Journal of Research in Biology

Journal of Research in Biology
An International Online Open Access

Original Research Publication group
Acute toxicity of insecticide, Diazinon and fungicide, Tilt (Propiconazole) on Pacific

white Shrimp, Litopenaeus vannamei postlarvae and Palaemon adspersus juveniles
Authors: ABSTRACT:
Majid Mohammad Nejad
Shamoushaki. Acute toxicity of Diazinon and Tilt (Propiconazole) agricultural toxins was
studied on Litopenaeus vannamei postlarvae and Palaemon adspersus juveniles,
aiming to determine the 50% lethal concentration (LC 50) in 96 h time duration. The
Institution: experiments were conducted at static condition and standard method in four days.
Department of Fishery, Physical and chemical factors were controlled through the experiment so that the
Bandar Gaz Branch,
amount of dissolved oxygen was fixed on 8 mg/L, temperature: 25 1 0C, pH: 7.5 to 8
Islamic Azad University,
and salinity: 31 ppt. The results showed that the mean LC50 values of Diazinon at 24,
Bandar Gaz, Iran.
48, 72 and 96 h were 0.298, 0.255, 0.237, 0.226 mg/L, and the mean LC50 values of
Tilt at 24, 48, 72 and 96 h were 9.021, 4.227, 4.032, 3.635 mg/L, to the Litopenaeus
Corresponding author: vannamei postlarvae. Also, the results showed that the mean LC50 values of Diazinon
Majid Mohammad Nejad at 24, 48, 72 and 96 h were 0.391, 0.330, 0.294, 0.277 mg/L, and the mean LC50
Shamoushaki.
values of Tilt at 24, 48, 72 and 96 h were 0.789, 0.763, 0.674, 0.611 mg/L, to the
Palaemon adspersus juveniles.
Email: Keywords:
Majid_m_sh@bandargaziau.ac.ir. Acute toxicity, Diazinon, Tilt, Litopenaeus vannamei, Palaemon adspersus.
Web Address: Article Citation:

http://jresearchbiology.com/ Majid Mohammad Nejad Shamoushaki.
Documents/RA0191.pdf.
Acute toxicity of insecticide, Diazinon and fungicide, Tilt (Propiconazole) on Pacific
white Shrimp, Litopenaeus vannamei postlarvae and Palaemon adspersus juveniles.
Journal of research in Biology (2012) 3: 160-166
Dates:
Received: 26 Jan 2012 /Accepted: 06 Feb 2012 /Published: 07 Mar 2012
Ficus Publishers.
This Open Access article is governed by the Creative Commons Attribution License (http://
creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-
commercial, distribution, and reproduction in all medium, provided the original work is properly
cited.
160-166 | JRB | 2012 | Vol 2 | No 3

Journal of Research in biology
Submit Your Manuscript
An International Open Access Online
Research Journal www.ficuspublishers.com www.jresearchbiology.com
Mohammad Nejad Shamoushaki, 2012
INTRODUCTION the household environment (Cox, 1992). Trade names for

White shrimp Litopenaeus vannamei (Boone) is Diazinon include Knox-out, Dianon and Basudin (EPA,
distributed throughout the Pacific coast from the Gulf of 2004). Tilt, with the chemical name of Propiconazol, is a
California to northern Peru. It is the major species of systemic fungicide, which in high usage is against
penaeid shrimp in the east hemisphere and contributes various kinds of rice diseases, such as stem rottenness
30% of farmed production of penaeid shrimp in the and etc (Mohammad Nejad Shamoushaki, 2005).
world (Perez Farfante and Kensley, 1997). This species Golestan province is a great pole of agriculture in the
inhabits wide ranges of salinity, including brackish water north of Iran and above 1.5 millions hectares of
of 12 ppt and saline water of 40 ppt (Menz and Blake, agricultural fields in areas are specified to cultivate
1980). Gomishan region in Golestan province is one of various kinds of farming products and dry farming. Thus
the main areas for culturing the Litopenaeus vannamei in different kinds of chemical fertilizers and vegetable
Iran. Distribution areas of Palaemon adspersus, Rathke pesticides are highly in use in Golestan. From a total
include the North Sea, Baltic Sea, Eastern Atlantic, degree of around 35000 metric tons of vegetable
Mediterranean, and Black Sea (Udekem dAcoz, 1999; pesticides, which are distributed in the Iran, around
Janas et al., 2004). Palaemon adspersus inhabits mainly 25000 metric tons of agricultural toxins are used by
Zostera covered bottoms only (Berglund, 1980; Baden farmers in agricultural fields of Golestan province
and Pihl, 1984). Also, this prawn exist in Caspian sea (Mohammad Nejad Shamoushaki, 2005). Toxicology of
and Gomishan lagoon, Iran. Pesticide use causes serious environmental parameters on Litopenaeus vannamei
environmental problems, especially in the dry season mainly focused on ammonia, nitrite, and some heavy
when the dilution capacity of the water systems is low, metal ions (Li et al, 2008), while there is no report on the
increasing the risk of high concentrations of toxic toxicology of Diazinon and Tilt to this species and
chemicals. Moreover, the dry season is often the critical Palaemon adspersus. As Gomishan area in Golestan
period for many animals, especially aquatic animals such province is only reproducing western white shrimp in the
as: fish and shrimp (Adedeji et al., 2009). Pesticide usage northern part of Iran, the acute toxicity pesticides
all over the world has increased dramatically during the Diazinon and Tilt, which is used a lot in Golestan
past few decades, coinciding with changes in farming district, on the Litopenaeus vannamei poslarvae and
and intensive agriculture practices. Hence the Palaemon adspersus juvenile were studied.
environmental pollution caused by pesticides, especially
in aquatic ecosystems, has become a grave problem MATERIALS AND METHODS
(Chilke, 2012). Direct or indirect contamination of water Shrimp and chemical supply
by pesticides can lead to fish and shrimp deaths, reduced To determine the acute toxicity of Diazinon and
productivity or elevated concentrations of undesirable Tilt from Litopenaeus vannamei postlarvae and
chemicals in edible fish tissue, which can affect the Palaemon adsperus from Litopenaeus vannamei
health of humans eating these aquatic animals (Adedeji postlarvae and Palaemon adspersus of Gomishan shrimp
et al., 2000). The organophosphate insecticide Diazinon brooding and culture center in Golestan province (north
(O,O-diethyl O-[6-methyl-2-(1-methylethyl)- 4- of Iran). In several processes some of these postlarvae
pyrimidinyl] phosphorothioate) has agricultural and (PL=20) and palaemon adspersus (1-2 gr) have been
commercial uses and it is used to control a variety of transferred to tanks to adapt to the new condition for 5-7
insects, primarily aphids, beetles, scales and pill bugs, in days. The experiments were carried out in 20 lit
161 Journal of Research in Biology (2012) 3: 160-166
aquariums (10 shrimps for each aquarium) with static Finally, LC10, LC50 and LC90 values at 24 48, 72 and 96
condition based on O.E.C.D method (TRC, 1984) with h, the maximum allowable concentration (MAC) value
five treatments and one blank with three repetitions. (LC50 in 96 h divided by 10) (TRC, 1984), the degree of
Physical and chemical factors were controlled through toxicity, mean of ineffective concentration (LOEC
the experiment. Dissolved oxygen was fixed on 8 mg/L, (Lowest Observed Effect Concentration)) (Finney,
temperature: 25 1 0C, pH: 7.5 to 8 and salinity: 31 ppt. 1971), of Diazinon and Tilt to Litopenaeus vannamei
Acute toxicity tests poslarvae and Palaemon adspersus juvenile were
The first experiment was conducted to determine determined.
the effects of acute toxicity (LC50 in 96 h) of Diazinon
(60 EM) and Tilt in two groups (Litopenaeus vannamei RESULTS AND DISCUSSION
postlarvae and Palaemon adspersus juvenile). For this The results showed that the mean LC50 values of
purpose, five treatments and one blank were used to test Diazinon at 24, 48, 72 and 96 h were 0.298, 0.255, 0.237,
toxicity; each treatment had three replications and 10 0.226 mg/L, and the mean LC50 values of Tilt at 24, 48,
shrimps per tank with 20 litres water capacity. Mortality 72 and 96 h were 9.021, 4.227, 4.032, 3.635 mg/L, to the
records were taken every 24 h (24, 48, 72, 96 h). Litopenaeus vannamei postlarvae (Table.1 and table. 2).
Movements and behaviors of the shrimps were Also, MAC value of Diazinon and Tilt were determined
investigated at the time of experiments. Finally, after to be 0.0226 and 0.3635 mg/L to Litopenaeus vannamei
early experiments, Diazinon concentration on postlarvae. The results showed that the mean LC50
Litopenaeus vannamei postlarvae was determined to be values of Diazinon at 24, 48, 72 and 96 h were 0.391,
0.15 - 0.3 mg/L and 2 - 8 mg/L, concentration of Tilt can 0.330, 0.294, 0.277 mg/L, and the mean LC50 values of
affect on Litopenaeus vannamei postlarvae. Then, based Tilt at 24, 48, 72 and 96 h were 0.789, 0.763, 0.674,
on this experiments LC10, LC50 and LC90 in 24, 48, 72 0.611 mg/L, to the Palaemon adspersus juveniles
and 96 h were measured. Also, early experiments (Table.3 and table. 4). Also, MAC value of Diazinon
showed 0.2 - 0.4 mg/L concentration of Diazinon and 0.4 and Tilt were determined to be 0.0277 and 0.0611 mg/L
- 1 mg/L concentration of Tilt can affect Palaemon to Palaemon adspersus juveniles, respectively. Also,
adspersus juvenile. Then LC10, LC50 and LC90 in 24, 48, LOEC (Lowest Observed Effect Concentration) which is
72 and 96 h of Diazinon and Tilt were measured for called LC10 in 96 h for Diazinon and Tilt were
Palaemon adspersus juvenile. determined 0.149 and 0.244 mg/L, respectively to
Statistical Analysis Litopenaeus vannamei and to Palaemon adspersus were
After obtaining the final results, the information determined 0.212 and 0.377 mg/L, respectively. The
was analysed statistically by probit program version 1.5 results showed that how pesticides tested concentration
(USEPA, 1985) and mortality was assessed at 24, 48, 72, increases, shrimps died in less time. In fact, for 24 hours
and 96 h after the early and dead shrimps removal. the mortality of shrimp in the amount of toxin is needed
Table1. Acute toxicity of Diazinon in 96 h on Table2. Acute toxicity of Tilt in 96 h on

Litopenaeus vannamei postlarvae Litopenaeus nnamei postlarvae
Concentration
Concentration 24 h 48 h 72 h 96 h 24 h 48 h 72 h 96 h
(mg/L)
LC10 0.2 0.176 0.158 0.149 LC10 3.162 1.448 1.375 1.244
LC50 0.298 0.255 0.237 0.226 LC50 9.021 4.227 4.032 3.635
LC90 0.443 0.370 0.360 0.342 LC90 25.730 12.340 11.825 10.621
Journal of Research in Biology (2012) 3: 160-166 162

Table3. Acute toxicity of Diazinon in 96 h on Table4. Acute toxicity of Tilt in 96 h on

Palaemon adspersus juvenile Palaemon adspersus juvenile
Concentra- Concentration
24 h 48 h 72 h 96 h 24h 48h 72h 96h
tion (mg/L) (mg/L)
LC10 0.275 0.243 0.214 0.212 LC10 0.539 0.525 0.447 0.377
LC50 0.391 0.330 0.294 0.277 LC50 0.789 0.763 0.674 0.611
LC90 0.557 0.447 0.404 0.362 LC90 1.157 1.066 1.018 0.989
more than 96 hours. Also, according to Table 5 studied by Li et al, (2008), while there is no report on the
(determination of toxicity in different pesticides) toxicology of Diazinon and Tilt to Palaemon adspersus.
Diazinon has high toxicity and Tilt has medium toxicity The 96 h LC50 values of ammonia-N on L. Vannamei
to Litopenaeus vannamei. Also, Diazinon and Tilt have juveniles were 24.39 mg/L at 15ppt; 35.4 mg/L at 25ppt;
high toxicity to Palaemon adspersus. The result of 39.54 mg/L at 35ppt, respectively. The 96 h LC50 values
Shrimp behaviors in different concentrations of toxin of NH3-N (un-ionized ammonia as nitrogen) were 1.20
showed that both shrimps, in high concentration of mg/L at 15ppt; 1.57 mg/L at 25ppt; 1.60 mg/L at 35ppt,
Diazinon and Tilt, had fast swimming, permanently then respectively (Lin and Chen, 2001). The 96 h LC50 value
got tired and died. While in low concentrations, during of nitrite-N on L. vannamei juveniles was 76.5 mg/L at
the first hours, there were no obvious reactions, then 15ppt, 178.3 mg/L at 25ppt, 321.7 mg/L at 35ppt (Lin
gradually faint. The main effect of concentrations of and Chen, 2003). The 96 h LC50 values of boron were
toxins was nervous system disorder, other external signs 25.05 mg/L at 3.0 ppt and 80.06 mg/L at 20.0ppt for
such as: imbalance, spiral swimming, and skin darkening Litopenaeus vannamei (Li et al, 2008). Acute toxicity of
were recorded to Litopenaeus vannamei postlarvae and ozone-produced oxidants (OPO) to juvenile Pacific white
Palaemon adspersus juvenile. So far no study has been shrimp, Litopenaeus vannamei, was assessed and found
reported on the effects of Diazinon and Tilt feature and to be 0.50 mg/L (Schroeder et al, 2010). Also, the results
other pollutants on Litopenaeus vannamei and Palaemon of this study and its comparison to the literature finally
adspersus in Iran. This could be due to the recent entry showed that the range of sensitivity to agricultural toxins
of Litopenaeus vannamei into Iran and there are still (e.g: Diazinon and Tilt), is more than that to the
many studies on this shrimp that can be made by ammonia and nitrite toxicity. But as to the pesticide
researchers. But since there is little information on the Diazinon there has been many studies on different
toxic effects of agricultural toxins to crustaceans, further species of fish. In other studies, 96 h LC50 value of
studies should be conducted to understand the toxic Diazinon were determined for the following species:
effect and mechanisms of agricultural toxins on Anguilla Anguilla: 0.08 mg/L, Blue gill: 0.46 mg/L,
crustaceans. As far as we know, agricaultural toxin such Fathead minnows (Pimephales promelas): 7.8 mg/L and
as Diazinon and Tilt can rapidly be accumulated in the zebra fish (Brachydanio rerio): 2.12 mg/L (Ansari et al.,
water and reach toxic concentrations via several
Table5. Determination of toxicity in different
processes, such as agricultural chemicals, laundry pesticides (Piri Zirkoohi and Orfog, 1997)
products, irrigation drain water, mining and processing, LC50 (mg/L) Degree of toxicity
and coal burning, so it presents a danger to aquatic Up to 100 Nearly no poison
10-100 toxicity Low
organisms (Li et al, 2008). Toxicology of environmental
1-10 toxicity Medium
parameters on Litopenaeus vannamei mainly focused on 0.1-1 toxicity High
ammonia, nitrite, and some heavy metal ions were Less to 0.1 toxicity Very high
1987). Also, 96 h LC50 value of Diazinon were and Department of Fishery, Bandar Gaz Branch, Islamic
determined for silver carp (Hypophthalmichthys molitrix) Azad University, Bandar Gaz, Iran. Also, With affection
and Abramis brama, 1.9 mg/L and 8.1 mg/L, respectively and deep appreciation we acknowledge indebtedness to
(Nasri Tajan, 1996); Acipenser persicus: 4.38 mg/L our friends and colleagues: Engineer Abdollatif Eimeri,
(Pajand, 1999); Acipenser nudiventris: 4.6 mg/L Engineer Ali Akbar Passandi, Engineer Voshtaie,
(Khoshbavar-Rostami and Soltani, 2002); Acipenser Engineer Saghali, Engineer Kia and Mr. Jalaly.
gueldenstadtii: 6.09 mg/L (Soltani and Khoshbavar-
Rostami, 2002); Huso huso: 4.99 mg/L (Khoshbavar- REFERENCES
Rostami et al., 2004); Acipenser stellatus: 4.98 mg/L Adedeji OB, Taiwo VO and Agbede SA. 2000.
(Khoshbavar-Rostami et al., 2005); grass carp Comparative haematology of five Nigerian freshwater
(Ctenopharyngodon idella): 15.13 mg/L (Pourgholam et fish species. Nig. Vet. J., 21:75-84.
al., 2006); Silurus glanis: 4.142 mg/L (Kprc et al.,
Adedeji OB, Adeyemo OK and Agbede SA. 2009.
2006); African catfish (Clarias gariepinus): 6.6 mg/L
Effects of diazinon on blood parameters in the African
(Adedeji et al., 2009), Bufo regularis: 0.44 mg/L
catfish (Clarias gariepinus), African Journal of
(Lawrence and Isioma, 2010). The results of this study
Biotechnology 8(16):3940-3946.
and its comparison to the literature finally show that
range of sensitivity to diazinon toxicant is, as shown Ansari BA, Aslam M and Kumar K. 1987. Diazinon
below: Anguilla anguilla > Litopenaeus vannamei > toxicity: activities of acetylcholinesterase and
Palaemon adspersus> Bufo regularis > blue gill> phosphatases in the nervous tissue of Zebra Danio,
Hypophthalmichthys molitrix > Brachydanio rerio > Brachydanio rerio (Cyprinidae). Acta Hydrochim.
Silurus glanis> A. persicus > A nudiventris >A. stellatus Hydrobiol., 15:301-306.
> Huso huso> A. gueldenstaedtii > Clarias gariepinus >
Baden SP, Pihl L. 1984. Abundance, biomass and
Pimephales promelas > Abramis brama >
production of mobile epibenthic fauna in Zostera marina
Ctenopharyngodon idella
(L.) meadows, western Sweden. Ophelia 23:65-90.
Also, 96 h LC50 values of Tilt was determined
as 3.9 mg/L at A nudiventris (Mohammad Nejad Berglund A. 1980. Niche differentiation between two
Shamoushaki, 2005). littoral prawns in Gullmar Fjord, Sweden: Palaemon
adspersus and P. squilla. Holarctic Ecology 3:111-115.
CONCLUSION
Cox C. 1992. Diazinon fact sheet. J. Pestic. Reform
The survey results indicated that the Diazinon
12:30-35.
and Tilt are very toxic to Litopenaeus vannamei and
Palaemon adspersus and endangers their health. EPA (Environmental Protection Agency). 2004.
Therefore, the shrimp farms in the vicinity of the Interim Reregistration Eligibility Decision, Diazinon.
pesticide use are in danger. In fact, L. vannamei shrimp United States Environmental Protection Agency, Office
farms in the vicinity of the use of pesticides can cause of Prevention, Pesticides and Toxic Substances 7508C.
harm to their health and affect the shrimp industry.
ACKNOWLEDGEMENT Finney D. 1971. Probit analysis. Cambridge Univ, Press
This research was supported by the Golestan 1-222 .
Province Fishery Organization, Gorgan, Golestan, Iran

Khoshbavar-Rostami HA and Soltani M. 2002. Acute salinity levels. Aquaculture 224:193-201.

toxicity of diazinon on Acipenser nudiventris. Second
Menz A and Blake BF. 1980. Experiments on the
Symposium on sturgeons. Rasht, Iran, Book of Abstracts
growth of Penaeus vannamei Boone. J. Exp. Mar. Biol.
72-75. (In Persian).
Ecol., 48:99-111.
Khoshbavar-Rostami H, Soltani M and Hassan
Mohammad Nejad Shamoushaki M. 2005. Accute
HMD. 2004. Acute toxicity and some hematological and
Toxicity of heavy metals (Pb, Cd and Zn) and
biochemical changes in giant sturgeon (Huso huso)
agricultural toxins (Diazinon, Hinosan and Tilt) LC50 in
exposed to diazinon. Bull. Eur. Assoc. Fish Pathol., 24
96 h to Acipenser nudivenrtis. MSc Thesis of Lahijan
(2):92-99.
Islamic Azad University. 1-4. (In Persian).
Khoshbavar-Rostami H, Soltani M and Yelghi S.
Perez Farfante I and Kensley B. 1997. Penaeid and
2005. Effects of diazinon on the hematological profiles
sergestoid shrimps and prawns of the world: keys and
of Acipenser stelletus and determination of LC50. J.
diagnoses. Memoires du Museum National DHistoire
Agric. Sci. Nature. Resure 12(5). (In Persian).
Naturelle, Paris. 233.
Kprc SO, Kprc K, Mevlt S, Ispir EU and Pala
Nasri Tajan M. 1996. Determining of LC50 in 96 h
M. 2006. Acute toxicity of organophosphorous pesticide
Granol diazinon 5% and emulsion 60% to Wetland
diazinon and its effects on behavior and some
Anzali Abramis brama. MSc Thesis of Lahijan Azad
hematological parameters of fingerling European catfish
University. Lahijan, Iran: 9-20. (In Persian).
(Silurus glanis), Pesticide Biochemistry and Physiology
86:99-105. Pajand Z. 1999. Acute toxicity of diazinon on
A.Stellatus and A. persicus Fingerlings. MSc Thesis of
Janas U, Zanzycki T, Kozik P. 2004. Palaemon
Lahijan Azad University. Lahijan. Iran. 45-60. (In
elegans a new component of the Gulf of Gdansk
Persian).
macrofauna. Oceanologia 46:143-0146.
Piri Zirkoohi M and Orfog V. 1997. Effect of some
Lawrence E and Isioma Tongo. 2010. Acute toxic
pesticides Commonly in Iranian agriculture on aquatic
effects of Endosulfan and Diazinon pesticides on adult
food chain. Tesis for pH-D degree submitted for the
amphibians (Bufo regularis). Journal of Environmental
academy of agricultural Sciences Godollo- Hungary. 1-
Chemistry and Ecotoxicology 2(5):73-78.
31.
Li E, Xiong Z, Chen L, Zeng C and Li K. 2008. Acute
Pourgholam R, Soltani M, Hassan DM, Ghoroghi A,
toxicity of boron to juvenile white shrimp, Litopenaeus
Nahavandi R and Pourgholam H. 2006. Determination
vannamei, at two salinities. Aquaculture 278:175-178.
of diazinon LC50 in Grass carp (Ctenopharyngodon
Lin YC and Chen JC. 2001. Acute toxicity of ammonia idella) and the effect of sublethal concentration of toxin
on Litopenaeus vannamei Boone Juveniles at different on some hematological and biochemical indices. Iranian
salinity levels. J. Exp. Mar. Biol. Ecol., 259:109-119. Journal of Fisheries Sciences 5(2):67-82.
Schroeder JP, Grtner A, Waller U and Hanel R.

Lin YC and Chen JC. 2003. Acute toxicity of nitrite on
2010. The toxicity of ozone-produced oxidants to the
Litopenaeus vannamei (Boone) juveniles at different
Pacific white shrimp Litopenaeus vannamei. Aquaculture
305:6-11.
Soltani M and Khoshbavar-Rostami H. 2002. The

study effects of diazinon on the some hematological and
biochemical changes of Acipeneser guldenstadti. Journal
of Marine Sceiences and Tecgnology 4(1):65-75. (In
Persian).
TRC. 1984. O.E.C.D. Guidelines for testing of

chemicals. Section 2. Effects on biotic systems 1-39.
Udekem d Acoz CD. 1999. Inventaire et distribution

des crustaces decapodes de lAtlantique nord-oriental,
de la Me diterranee et des eaux continentals adjacentes
au nord de 25_N. Collection Patrimoines Naturels.
Museum national dHistoire naturelle, Paris. 383.
USEPA (United States Environmental Protection

Agency). 1985. Methods for measuring the acute
toxicity of effluents to freshwater and marine organisms.
3rd Ed. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory,
Cincinnati, OH. EPA-600/4-85/013.
Submit your articles online at Ficuspublishers.com

Advantages
Easy online submission
Complete Peer review
Affordable Charges
Quick processing
Extensive indexing
Open Access and Quick spreading
You retains your copyright
submit@ficuspublishers.com
www.ficuspublishers.com/submit1.aspx.

Growth responses of petroleum refinery effluent bacteria to phenol

Authors: ABSTRACT:
Nwanyanwu CE, Nweke
CO and Orji JC. Studies of the fungal and bacterial population of petroleum refinery effluent
samples were carried out by microbial enumeration and determination of growth
responses of bacterial isolates in increasing doses (0 - 15mM) of phenol in mineral salt
-phenol agar. The total aerobic heterotrophic bacterial count ranged from (2.05 0.4)
x 107 to (2.52 0.5) x 108 CFU/ml, total phenol-utilizing bacteria ranged from (1.18
0.3) x 106 to (1.02 0.3) x 107 CFU/ml and the total fungal count ranged from (3.1
Institution:
Department of 1.3) x 103 to (3.9 0.5) x 104 CFU/ml in the effluent samples. Bacillus sp. RWW,
Microbiology, Federal Aeromonas sp. RBD, Escherichia coli OPWW and Staphylococcus sp. DP were isolated
University of Technology, from the samples. Growth responses of the isolates on increasing doses of phenol in
P.M.B. 1526, Owerri, mineral salt-phenol agar showed that Bacillus sp. RWW and Escherichia coli OPWW
Nigeria. had highest growth on 0.5mM ( 47.06mg/l) while Aeromonas sp. RBD and
Staphylococcus sp. DP had their highest growth on 1.0mM ( 94.11mg/l). Bacillus sp.
RWW and Escherichia coli OPWW showed least growth on 15.0mM (1,412mg/ml) of
phenol. Escherichia coli OPWW exhibited highest growth in mineral salt broth
containing 11.0mM of phenol with OD540nm of 0.324 in 144 h resulting in the fastest
utilization of phenol for growth. The highest specific growth rate of 0.013 h -1 at 11 mM
Corresponding author: (1,035mg/l) of phenol was obtained for Bacillus sp. RWW and Escherichia coli.
Nwanyanwu CE Staphylococcus sp. DP had the lowest specific growth rate of 0.011 h -1 at 11 mM of
phenol. These bacterial strains could be considered phenol-resistant and are
potentially applicable in the removal of phenolic compounds from contaminated
environmental media.
Email: Keywords:
cnwanyanwu2000@yahoo.com Growth responses, phenol-utilizing bacteria, refinery effluent.

http://jresearchbiology.com/ Nwanyanwu CE, Nweke CO and Orji JC.
Documents/RA0185.pdf. Growth responses of petroleum refinery effluent bacteria to phenol.
Dates:
Received: 09 Jan 2012 /Accepted: 18 Jan 2012 /Published: 13 Mar 2012
Ficus Publishers.
cited.
167-177 | JRB | 2012 | Vol 2 | No 3

Nwanyanwu et al.,2012
INTRODUCTION Industrial effluent discharges have led to the

The utilization of chemicals as carbon or energy pollution of aquatic and terrestrial environments over the
sources by living cells is basic to all forms of life. years. Many mesophilic microbial species have been
Adaptation of living cells over the centuries to consume implicated in the degradation of phenol leading to a
the natural biochemicals found on earth is the generally successful bioremediation of phenol-contaminated
accepted narrative, but the significantly different organic environments. Improvement of microbial efficiency has
species which are produced by man have led to been directed towards the utilization of microbial seeding
environmental problems due to the resistance or as a means of controlling industrial effluent discharges
complete recalcitrance to mineralization by any living (Idise et al., 2010). Biodegradation enables organic
species (Hill et al, 1996; Khleifat, 2006). pollutants to be converted from harmful organic
Phenol and its derivatives are common substances into harmless inorganic substances. The
constituents of industrial effluents such as ability of microorganisms to degrade phenol and the rate
pharmaceutical, industrial resin manufacturing, of phenol degradation is governed by certain factors such
pharmaceutical industries, plastic, coke oven plants, as oxygen tension, temperature, pH, and microbial
petrochemical and petroleum refineries (Charest et al., abundance as well as nutrient availability (Zhao and Zhu,
1999; Ruiz-Ordaz et al., 2001; Jindrova et al., 2002; 1997). The type and number of microbial species
Rigo and Alegre, 2004). Phenol is toxic to all forms of involved in degradation may influence the rate and
life even at low concentrations and causes taste and extent of the process (Atlas, 1981; Leahy and Colwell,
odour problems in drinking water (Boszczyk-Maleszak 1990). Despite the presence of microorganisms capable
et al., 2002; Monkiedje et al., 2004). The toxic action of of degrading the aromatic compounds, the concentration
phenol is always associated with the loss of cytoplasmic and type of the pollutant limit the biodegradation of the
membrane integrity. As a result of phenol membrane- inherent compounds. If the organic pollutant is not
damaging properties on microorganisms, cytoplasmic available to the microorganisms and the appropriate
membrane integrity is lost which in turn results in the enzymes do not come in contact with the substrate in the
disruption of energy transduction, disturbance of proper way, degradation will not occur (Atlas, 1981).
membrane barrier function, inhibition of membrane Biodegradation of phenol and other pollutants in the
protein function and subsequent cell death (Kewelo et natural ecosystem though complex has greatly enhanced
al., 1990; Heipieper et al., 1991; Yap et al., 1999). The the removal of harmful ions and solids from the
high-volume use of phenols world wide and their environment by transformation into readily usable and
potential toxicity has led to the inclusion of phenols on less/non toxic materials by microorganisms (Tanaka et
the list of priority pollutants (Veeresh et al., 2005). al., 1993). This research evaluated the growth responses
Owing to the importance of microbial activities in of bacteria isolated from petroleum refinery wastewaters
biogeochemical cycling, several biological parameters to various concentrations of phenol.
have been used to evaluate the toxic effect of phenol on
bacterial population which includes changes in cell MATERIALS AND METHODS
numbers, biomass measurement, specific and non Sample collection and characterization
specific enzymatic activities (Kuo and Genthner, 1996; Physicochemically treated raw effluent (addition
Nwanyanwu and Abu, 2010, Nweke and Okpokwasili, of additives, flocculation, sedimentation and filtration)
2010a,b; 2011). (RWW), biologically treated effluent [Rotary BioDisk,
(RBD)], observation pond treated effluent [Oxidation flasks. The flasks were incubated on a rotary shaker
pond, (OPWW)] and Discharge Pipe effluent (DP) incubator operating at 150 rpm for 24 h at room
samples of Port Harcourt petroleum refinery were temperature (28 2oC). Cells were harvested by
collected in sterile polyethylene containers. The centrifugation at 4000 rpm for 10 minutes. Harvested
containers were rinsed thrice with the effluent samples at cells were washed twice in sterile deionized distilled
the point of collection. To avoid deterioration, the water and resuspended in the same medium. The cell
samples were taken to the laboratory in icebox within six suspensions were standardized in a spectrophotometer to
hours of collection for the determination of pH, Total an optical density of 0.5 at 540 nm.
Hydrocarbon (THC), electrical conductivity, Biological Assay for the effect of phenol on the growth of
Oxygen Demand (BOD), Chemical Oxygen Demand bacteria in BH agar
(COD), Phosphate (PO4), Sulphate (SO4), phenol The phenol-utilizing bacterial isolates were used
(C6H5OH), Lead (Pb), Zinc (Zn), and Copper (Cu) for this assay. Aliquots of stock solution of phenol
following standard procedures (APHA, 1985). (20mM) were diluted with Bushnell and Haas (BH) broth
Enumeration of microorganisms medium (containing per litre: K2HPO4, 1.0g; KH2PO4,
Aerobic heterotrophic bacteria, phenol-utilizing 1.0g; NH4NO3, 1.0g; MgSO4.7H2O, 0.2; CaCl2.2H2O,
bacteria and fungal populations in the samples were 0.02; FeCl3.6H2O, 0.085g) in 250ml Erlenmeyer flasks to
enumerated on nutrient agar, mineral salt-phenol agar obtain different concentrations of phenol (0-15 mM).
(Hill and Robinson, 1975) and potato dextrose agar This was followed by the addition of 2% agar into the
plates respectively. In each medium, 0.1 ml aliquot of flasks. The flasks were sterilized by autoclaving at 121 oC
sample dilutions were spread-inoculated onto the agar for 15 min after which the contents were poured into
o
surface and incubated at room temperature (28 2 C) for sterile plates set up in triplicates. The plates were
24, 72 and 120 h for aerobic heterotrophic bacteria, allowed to solidify at room temperature and were, then
phenol-utilizing bacteria and fungi respectively. The dried in the oven at a temperature of 800C. Control flasks
discrete bacterial colonies that grew profusely on the that received the same treatment except phenol was set
mineral salt-phenol agar representing the preponderant up. An aliquot (0.1ml) of the standardized inoculum
morphotypes in their respective sources were purified on decimally diluted in physiological saline (0.85 % w/v
freshly prepared nutrient agar (Lab M) and stored in NaCl) were spread on the agar surface. All inoculated,
nutrient agar slant at 4oC. Characterizations of phenol- plates were incubated aerobically at 28 20C for 72 h,
utilizing bacteria were done using standard after which colonies were counted.
microbiological methods. Identification to the genus Growth and phenol utilization assay
level followed the schemes of Holt et al (1994). The Liquid cultures were conducted in order to
isolates were designated according to their sources examine more parameters such as phenol concentration
(RWW for Raw Waste Water, RBD for Rotary BioDisk, and growth of the bacterial strains. Hundred millilitre
OPWW for Observation Waste Water and DP for (100 ml) of BH medium was placed in a duplicate set of
Discharge Pipe). 250ml Erlenmeyer flasks. The flasks were supplemented
Preparation of inoculum and culture condition with aliquot of phenol (20 mM) to bring the final phenol
The phenol-utilizing bacterial strains used for the concentrations in the flasks to 2, 5, 8 and 11 mM. The
assay were grown in 100 ml of sterile nutrient broth flasks were sterilized by autoclaving, inoculated with the
media (HIMEDIA) contained in 250 ml Erlenmeyer test organisms upon cooling and incubated at 30 oC in an

incubator. At specific time, samples were withdrawn to degraders present in such specific habitats (Kopytko and
determine cell growth and remaining phenol. Controls Jacome, 2008). Total viable counts of aerobic
included cells in BH medium without phenol and the heterotrophic population were highest in RBD and
medium supplemented with phenol but without cells. lowest in RWW with microbial load of (2.52 0.5) x 10 8
This is to assess for endogenous respiration of the and (2.05 0.4) x 107 CFU/ml respectively. The
cultures as well as to measure abiotic loss of phenol difference in microbial load of the effluents may be
during the test period. At specific intervals of time attributed to differences in phenolic content. The
samples were collected and measured for cell growth and utilization of phenol in the rotary biodisk may have
phenol degradation. The cell growth was determined by resulted in the increased number of bacterial cells. The
measuring the Optical Density (OD) at 540 nm using nutrient content of the effluent samples measured as
spectrophotometer. The concentration of phenol in the Biological Oxygen Demand (BOD), Chemical Oxygen
medium was determined by the method of Folsom et al. Demand (COD), phosphate and nitrate may also be a
(1990). For each initial concentration of phenol, specific contributing factor in the microbial population obtained
-1
growth rate (, h ) was estimated using equation 1. (Ganapathiselvam et al., 2011). Total phenol-utilizing
bacteria were found to be highest in DP and lowest in
In X 2 X 1
1 OPWW with total load of (1.02 0.3) x 107 and (1.18
t 2 t1
0.3) x 106 CFU/ml respectively while total fungal load
Where X1 and X2 are the cell biomass (cell density)
was found to be highest in RBD and lowest in DP with
obtained at time t 1 and t2.
total load of (3.9 0.5) x 104 and (3.1 1.3) x 103 CFU/
Phenol degradation rate (Qs, mg/l.h) was
ml respectively.
determined from the maximum slope in plot of phenol
The bacterial isolates from the petroleum refinery
concentration (mg/l) versus time of incubation as
effluent samples were identified and designated as
described by Afzal et al. (2007).
Bacillus sp. RWW, Aeromonas sp. RBD, Escherichia
Statistical analysis
coli OPWW and Staphylococcus sp. DP comprising two
The statistical analysis was achieved using
Table 1: Physicochemical properties of petroleum
Microsoft Excel 2003. Biodegradation rate of phenol for refinery effluent samples
each bacterium was compared pairwise using students t- Sample sourcea
test with 5 % significance level OPW
Parameter/unit RWW RBD DP
W
pH 7.64 8.18 7.45
8.87
RESULTS AND DISCUSSION Temperature oC 26.4 26.1 26.8
26.7
Elect. Conduc.
The physicochemical properties and the 845 443 926 643
(scm-1)
microbial counts of the petroleum refinery effluent THC (mg/l) 17.5 15.0 21.0 16.0
BOD (mg/l) 32.0 8.0 12.8 12.8
samples are shown in Tables 1 and 2 respectively. The
COD (mg/l) 112.0 76.0 114.0 84.0
samples are slightly alkaline (pH 7.64 8.87). The PO4 (mg/l) 0.22 0.14 0.13 0.12
microbiological analysis of oil refinery effluents NO3 (mg/l) 2.60 1.20 1.80 1.20
Phenol (mg/l) 71.2 13.6 10.1 9,4
indicated that the effluents are a good habitat for bacteria Pb (mg/l) <0.01 <0.01 <0.01 <0.01
and fungi. The isolation of various bacterial genera from Zn (mg/l) 0.13 0.02 0.06 0.08
Cu (mg/l) <0.01 <0.01 0.01 0.01
the effluents that are able to utilize phenol as carbon and a
RWW =Raw Wastewater, RBD = Rotary Biodisk,
energy sources showed that they are potential phenol OPWW = Observation Pond Wastewater,
DP = Discharge Pipe
Table 2: Microbial load of petroleum refinery effluent samples

Count (CFU/ml)
a Total aerobic Total phenol-utilizing
Sample source
heterotrophic bacteria bacteria Total fungi
RWW (2.05 0.4) x 107 (1.24 0.4) x 106 (4.1 1.2) x 103
RBD (2.52 0.5) x 108 (1.31 0.3) x 106 (3.9 0.5) x 104
OPWW (2.76 0.3) x 107 (1.18 0.3) x 106 (3.6 0.5) x 103
DP (2.50 1.2) x 108 (1.02 0.3) x 107 (3.1 1.3) x 103
a
RWW =Raw Wastewater, RBD = Rotary Biodisk, OPWW = Observation Pond
Wastewater, DP = Discharge Pipe
each of Gram negative and Gram positive organisms that phenol concentration of 1.0mM ( 94.11mg/l) with a
exhibit a great deal of metabolic versatility. The toxicity total viable counts of (7.5 0.3) x 106 and (7.3 0.4) x
of phenol to the isolate was assessed using population 106 CFU/ml respectively. This implies a dose-dependent
growth on phenol-amended mineral salt medium. Figure population growth response of the isolates to the
1 showed the effect of phenol on the population of the concentrations of phenol in the growth medium as
test isolates growing on increasing doses of phenol compared to the controls with growth population values
ranging from 0.05 to 15.0mM ( 4.71 -1,411.7 mg/l). At of (2.1 0.4) x 105, (5.7 0.2) x 105, (3.3 0.3) x 105
0.5 mM ( 47.06mg/l) phenol, Bacillus sp. RWW and and (3.3 0.5) x 105 CFU/ml for Bacillus sp. RWW,
Escherichia coli OPWW have their highest growth with Aeromonas sp. RBD, Escherichia coli. OPWW and
6
total viable counts of (7.1 0.5) x 10 and (7.9 0.2) Staphylococcus sp. DP respectively. The growth of
6
x10 CFU/ml respectively while Aeromonas sp. RBD bacteria in the control medium (without phenol) may be
and Staphylococcus sp. DP have their highest growth at as a result of organic impurities present in the medium
Total viable count (CFU/ml x 106)
Phenol (mM)
Figure 1: Growth of Bacillus sp. RWW (a); Aeromonas sp. RBD (b); Escherichia coli. OPWW (c) and
Staphylococcus sp. DP (d) on Bushnell-Haas agar medium amended with increasing doses of phenol.

components, which are utilized by the organisms as toxicity at such high concentration leading to incomplete
carbon and energy source. The total viable counts of the utilization of phenol in the medium (Acua-Argelles et
organisms progressively decreased as the phenol al., 2003; Ruiz-Ordaz et al., 1998). Also, studies have
concentration increases. At 15.0mM ( 1,411.7 mg/l) shown that lower doses of phenol are more readily
Bacillus sp. RWW and Escherichia coli OPWW were utilized than higher doses. This corroborate with the high
highly inhibited while Aeromonas sp. RBD and growth responses at low doses (0.5 and 1.0 mM
Staphylococcus sp. DP showed little population growth respectively) of phenol. Stimulation of dehydrogenase
5 5
of (1.0 0.01) x 10 and (2.0 0.1) x 10 CFU/ml activity at low dose of phenol was reported for Bacillus
respectively. The inhibitory nature of phenol at high and Pseudomonas species isolated from petroleum
concentrations is well known, and has been reported by refinery wastewater (Nweke and Okpokwasili, 2010a, b).
many authors (Kotturi et al., 1991; Margesin et al., 2004; Stimulation of dehydrogenase activity in some bacteria
Khleifat, 2006; Okpokwasili and Nweke, 2006). by phenol could indicate the use of phenol as a growth
Although phenol was toxic to the organisms, substrate. Similar stimulation and inhibition of
they could utilize it as source of carbon and energy at dehydrogenase activity in soil Acinetobacter species by
low concentrations. The growth patterns of the phenol- phenol derivatives at low and high concentrations
utilizing bacteria in BH medium containing different respectively was reported by Okolo et al. (2007). The
concentrations of phenol presents an interesting progressive inhibition of dehydrogenase activity with
observation. The results conform to the work of Marrot increasing concentration of phenols is in line with the
et al. (2008) that reported microbial responses to phenol well documented inhibitory nature of phenols at high
at lower concentrations, depicting an increase in concentrations even for organisms which can use phenols
microbial biomass. Depression of growth of Bacillus sp. as growth substrates (Acua-Argelles et al., 2003; Ruiz-
RWW and Escherichia coli OPWW at 15mM ( 1,411.7 Ordaz et al., 1998).
mg/l) may be due to inactivation of enzymes by phenol Growth of the organisms in BH medium
Phenol (mM)
Time (h)
Figure 2: Biodegradation of different concentrations of phenol in mineral salt medium by the
bacterial strains. Values are mean of duplicate determination.

supplemented with different concentrations of phenol Satsangee and Ghosh (1990) that non acclimated
was studied. Phenol utilization was well correlated with organisms take longer period than acclimated ones to
growth. Utilization and growth of the bacterial strains in degrade organic compounds. Also, Joseph and Joseph
different concentrations of phenol are shown in Figures (1999) reported that phenol toxicity depends on the
2 and 3 respectively. Phenol was completely utilized by sensitivity as well as pre-exposure of organism to the
the isolates within 180 h. Two millimolar [(2.0 mM toxicant. At high concentrations, phenol has been
(188.2 mg/l)] of phenol was utilized completely within observed to inhibit microbial growth even to organisms
60 h by Bacillus sp. RWW and Escherichia coli. OPWW that utilize it as sole source of carbon and energy. This is
while same concentration of phenol was utilized in line with the reports of Collins and Daugulis (1997)
completely within 72 h by Aeromonas sp. RBD and and Nwanyanwu and Abu (2011) who observed the toxic
Staphylococcus sp. DP. Staphylococcus sp. DP effect of phenol at the membrane level, thereby
completely degraded 11.0 mM ( 1,035.2 mg/l) in 180 h disrupting the activity of enzymes in phenol-utilizing
(Figure 2). Time-dependent degradation of organic bacteria.
compounds has been reported to be linked with The biomass concentration profile of the pure
concentration of the organic compound as observed by cultures expressed as optical density (OD540nm) at
many authors (Colwell and Walker, 1977; Kotresha and different initial concentrations of phenol is shown in
Vidyasagar, 2008). This longer period of utilization of Figure 3. It was observed that the lag phases of growth
phenol by the organisms may be attributed to non- of the bacterial isolates were below 12 h. Highest cell
acclimation of the bacterial strains to increasing doses of densities (OD540nm) recorded were 0.341 at 11 mM and
phenol or substrate inhibition effect of phenol at high 0.182 at 2 mM phenol for Escherichia coli OPWW. This
concentrations. This corroborates the observation of resulted in the faster conversion rates of phenol for the
Optical density (OD540nm)
Time (h)
Figure 3: Growth of the bacterial strains on different concentrations of phenol in
mineral salt medium. Values are mean of duplicate determination.

bacterial growth compared to other pure cultures. specific growth rate of 0.15 h-1 (Bai et al., 2007).
Though, the other pure cultures showed robust Similarly, mixed microbial consortium grew at
-1
absorbance of cell growth in all phenol concentrations maximum specific growth of 0.37h (Saravanan et al.,
indicating that the cultures are capable of utilizing 2008), 0.143 h-1 (Nuhoglu and Yalcin, 2005) and 0.31 h-1
phenol much efficiently. Specific growth rates () for (Bajaj et al., 2009). However, the growth rates of
each initial phenol concentration (So) was calculated Escherichia coli, Aeromonas, Bacillus and
from the plot of ln (X) versus time in logarithmic phase. Staphylococcus species are similar to that of
The slope of the linear logarithmic plots of optical Pseudomonas aeruginosa and Pseudomonas
density during the exponential phase gives the specific pseudomallei degrading phenol in saline solutions (Afzal
growth rate. Specific growth rate of all the bacterial et al., 2007). The rate of phenol degradation of the
strains increased rapidly at phenol concentration of 2.0 organisms during their growth in phenol medium are
mM ( 188.22mg/l) and then progressively decreases shown in Table 3, The highest rate of phenol
with increase in substrate concentration suggesting degradation (9.16 mg/l.h) at 11 mM (1035 mg/l) was
phenol toxicity to the organisms (Figure 4). The highest observed with Escherichia coli OPWW. The degradation
specific growth rates obtained at phenol concentration of rates at 11 mM were higher than the degradation rates at
-1
11.0 mM ( 1035.21 mg/l) was 0.013 h for Bacillus sp. 2 mM. This indicates strong potential of these organisms
RWW and Escherichia coli OPWW while the lowest to degrade phenol. The slow rate of growth and
specific growth rate of 0.011 h-1 at 11.0mM was biodegradation may be attributed to the lack of
observed in Staphylococcus sp. DP. The Escherichia acclimation or decrease in the effective reactivity of the
coli, Aeromonas, Bacillus and Staphylococcus species enzyme system within the cell. Similar report had been
grew relatively at slow rate. The mean maximum specific observed by Yap et al., (1999) in their work using
growth of Acinetobacter species in phenol was reported Comamonas testosteroni strain P15 to eliminate phenol.
-1
to be 0.83 h (Hao et al., 2002). Ewingella americana They reported that the low degradation rate of phenol by
degrading 300mg/l phenol grew at maximum specific the organism was in order to counteract the adverse
1
growth rate of 0.32 and 0.29 h for starved and non- effects of phenol inhibition. The t-test showed that the
starved cells respectively (Khleifat, 2006). A phenol-
Table 3: Biodegradation rate (Qs) of refinery
degrading Alcaligenes faecalis grew at maximum effluent bactria under different initial phenol
concentrations
Biodegradation rate (mg/l.h)

Specific growth rate (h-1)
Phenol concentration (mg/l)a

Bacteria 188(2) 471(5) 753(8) 1035(11)
Bacillus sp.
4.49 6.18 11.63 8.22
RWW
Aeromonas
5.88 6.23 7.30 8.47
sp. RBD
Escherichia coli
OPWW
5.25 6.88 8.32 9.16
Staphylococcus
Phenol (mM), So sp. DP
3.61 5.63 6.06 6.33
Figure 4: Specific growth rate of the bacteria under values in parenthesis represent phenol
different initial phenol concentrations concentration in mM

biodegradation rate varied significantly (p < 0.05) among -cresol using Alcaligenes faecalis. Proc. Biochem.,
the test organisms. However there was no significant 42:510-517.
difference in the biodegradation rates of Aeromonas sp.
BoszczykMaleszak H, Chorazy M, Bieszkiewicz E,
RBD and Staphylococcus sp. DP as well as Escherichia
Kacieszczenko J. 2002. Phenol utilization by fungi
sp OPWW and Staphylococcus sp. DP.
isolated from activated sludge. Acta Microbiol. Polon.,
Although, they grew at low rate, the organisms
51:182-191.
demonstrated strong potential to utilize and degrade
phenol at high phenol concentrations of 11.0 mM Charest A, Bisaillon JG, Lpine F, Beaudet R. 1999.
(1,035.2 mg/l). This indicated that these strains have Removal of phenolic compounds from a petrochemical
remarkable potential for application in the treatment of effluent with a methanogenic consortium. Can. J.
phenolic wastewater and in the bioremediation of phenol Microbiol., 45:235-241.
-contaminated media.
Collins LD, Daugulis AJ. 1997. Characteristics and
optimization of a two-phase partitioning bioreactor for
REFERENCE
the biodegradation of phenol. Appl. Micobiol.
Acua-Argelles ME, Olguin-Lora P, Razo-Flores E.
Biotechnol., 48:18-22.
2003. Toxicity and kinetic parameters of the aerobic
biodegradation of phenols and alkyl phenols by a mixed Colwell RR, Walker JD. 1977. Ecological aspects of
culture. Biotechnol. Lett., 25:559-564. microbial degradation of petroleum in the marine
environment. Crit. Rev. Microbiol., 5:423-445.
Afzal M, Iqbal S, Rauf S and Khalid ZM. 2007.
Characteristics of phenol biodegradation in saline Folsom BR, Chapman PJ, Pritchard PM. 1990.
solutions by monocultures of Pseudomonas aeruginosa Phenol and trichloroethylene degradation by
and Pseudomonas pseudomallei. J. Hazard. Mat., 149:60 Pseudomonas cepcia GA: Kinetics and interaction
-66. between substrates. Appl. Environ. Microbiol., 56:1279-
1285.
APHA. 1985. Standard Methods for the Examination of
Water and Wastewater, 16th edition. American Public Hao O, Kim M, Seagreen E and Kim H. 2002.
Health Association, American water works Association Kinetics of phenol and chlorophenol utilization by
and Water Pollution control Federation, Washington, Acinetobacter isolates. Chemosphere 46(6):79-807.
D.C.
Heipieper HJ, Kewelo H, Rehm HJ. 1991. Influence of
Atlas RM. 1981. Microbial degradation of petroleum phenols on growth and membrane permeability of free
hydrocarbons: an environmental perspective. Microbiol. and immobilized Escherichia coli. Appl. Environ.
Rev., 45:180-209. Microbiol., 57:1213-1217.
Bajaj M, Gallart C, and Winter J. 2009. Phenol Hill GA, Milne BJ, Nawrock PA. 1996. Cometabolic
degradation kinetics of an aerobic mixed culture. degradation of 4-chlorophenol by Alcaligenes eutrophus.
Biochem Eng. J., 46(2):205-209. Appl Microbiol. Biotechnol., 46:163-168.
Bai J, Wen J-P, Li H-M and Jiang Y. 2007. Kinetics Hill GA, Robinson CW. 1975. Substrate inhibition
modelling of growth and biodegradation of phenol and m kinetics: phenol degradation by Pseudomonas putida.

Biotechnol. Bioeng., 17:1599-1615. Kuo CW, Genthner BRS. 1996. Effect of added metal
ions on biotransformation and biodegradation of 2-
Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams
chlorophenol and 3-chlorobenzoate in anaerobic bacterial
ST. 1994. Bergeys Manual of Determinative
consortium Appl. Environ. Microbiol., 62:2317-2323.
Bacteriology, 9th ed. Williams, Wilkins. Baltimore.
Leahy JG, Colwell RR. 1990. Microbial degradation of
Idise OE, Ameh JB, Yakubu SE, Okuofu CA. 2010.
hydrocarbons in the environment. Micrbiol. Rev., 54:305
Biodegradation of a refinery effluent treated with organic
-315.
fertilizer by modified strains of Bacillus cereus and
Pseudomonas aeruginosa. Afr. J. Biotechnol., 9 Margesin R, Bergauer P, Gander S. 2004. Degradation
(22):3298-3302. of phenol and toxicity of phenolic compounds: a
comparison of cold-tolerant Arthrobacter sp. and
Jindrova E, Chocova M, Demnerova K, Brenener V.
mesophilic Pseudomonas putida. Extremophiles 8:201-
2002. Bacterial aerobic degradation of benzene, toluene,
207.
ethylbenzene and xylene. Folia Microbiol., 47:83-94.
Marrot B, Barrios-Martinez A, Moulin P, Roche N.
Joseph V, Joseph A. 1999. Acclimation of algal species
2008. Biodegradation of high phenol concentration in a
following exposure to phenol. Bull. Environ. Contam.
membrane bioreactor. Int. J. Chem. React. Eng., 6.
Toxicol., 62:87-92.
Monkiedje A, Njine M, Meyabeme-Elono AL, Zebaze
Kewelo H, Weyrauch G, Rehm HJ. 1990. Phenol
SH, Kemka N, Chounwou PB, Djomo JE. 2004. Fresh
induced membrane changes in free and immobilized
water microcosms-based assessment of ecotoxicological
Escherichia coli. Appl. Microbiol. Biotechnol., 33:66-71.
effects of a chemical effluent from the pilcan industry in
Khleifat KM. 2006. Biodegradation of phenol by Cameroon. Int. J. Environ. Res. Publ. Health 1(2):111-
Ewingella americana: Effect of carbon starvation and 123.
some growth conditions. Proc. Biochem., 41:2010-2016.
Nwanyanwu CE, Abu GO. 2010. In vitro effects of
Kopytko M, Jacome LAP. 2008. Alternative for phenol petroleum refinery wastewater on dehydrogenase activity
biodegradation in oil contaminated wastewaters using an in marine bacterial strains. Ambi. Agua., 5:21-29.
adapted bacterial biofilm layer. Rud. Geo. Naft. Zb.,
Nwanyanwu CE, Abu GO. 2011. Assessment of
20:71-82.
viability responses of refinery effluent bacteria after
Kotresha D, Vidyasagar GM. 2008. Isolation and exposure to phenol stress. J. Res. Biol. 8: 594 602.
characterisation of phenol-degrading Pseudomonas
Nweke CO, Okpokwasili GC. 2010a. Influence of
aeruginosa MTCC 4996. World J. Microbiol.
exposure time on phenol toxicity to refinery wastewater
Biotechnol., 24:541-547.
bacteria. J. Environ. Chem. Ecotoxicol., 2(2):20-27.
Kotturi G, Robinson CW, Inniss WE. 1991. Phenol
Nweke CO, Okpokwasili GC. 2010b. Inhibition of
degradation by a psychrotrophic strain of Pseudomonas
dehydrogenase activity in petroleum refinery wastewater
putida. Applied Microbiology and Biotechnology 4:539-
bacteria by phenolic compounds. Ambi. gua., 5:6-16.
543.

Nweke CO, Okpokwasili GC. 2011. Inhibition of - Tanaka OH, Venkatswaran K, Komukai S, Toki H,
galactosidase and -glucosidase synthesis in petroleum Iwabuch T, Miyachi S. 1993. Ecodynamics of oil
refinery effluent bacteria by phenolic compounds. Ambi. degrading bacteria and significance of marine mixed
gua., 6(1):40-53. populations in the degradation of petroleum compounds.
International Oil Spill Conference 780-785.
Nuhoglu A, Yalcin B. 2005. Modelling of phenol
removal in a batch reactor. Proc. Biochem., 40:1233- Veeresh GS, Kumar P, Mehrotra I. 2005. Treatment of
1239. phenol and cresol in upflow anaerobic sludge blanket
(UASB) process: a review. Wat. Res., 39:154-170.
Okolo JC, Nweke CO, Nwabueze RN, Dike CU,
Nwanyanwu CE. 2007. Toxicity of phenolic compounds Yap LF, Lee YK, Poh CL. 1999. Mechanism for phenol
to oxidoreductases of Acinetobacter species isolated tolerance in phenol-degrading Comamonas testosteroni
from a tropical soil. Sci. Res., Essay 7:244-250. strain. Appl. Microbiol. Biotechnol., 51:833-840.
Okpokwasili GC, Nweke CO. 2006. Microbial growth Zhao J, Zhu J. 1997. Study on the Biodegradation of oil
and substrate utilization kinetics. Afr. J. Biotechnol., 5 spilled on the sea. International Oil Spill Conference 989
(4):305-317. -990.
Rigo M, Alegre RM. 2004. Isolation and selection of

phenol-degrading microorganisms from industrial
wastewaters and kinetics of the biodegradation. Folia
Microbiol., 49:41-45.
Ruiz-Ordaz N, Ruiz-Lagunez JC, Gonzalez JUC,

Manzano EH, Urbina EC, Mayer JG. 2001. Phenol
biodegradation using a repeated batch culture of Candida
tropicalis in a multistage bubble column. Rev. Latino-
Americana Microbiol., 43:19-25.
Ruiz-Ordaz N, Hernndez-Manzano E, Ruiz-Lagnez

JC, Cristiani-Urbina E, Galndez-Mayer J. 1998.
Growth kinetics model that describes the inhibitory and
lytic effects of phenol on Candida tropicalis yeast.
Biotechnol. Prog., 14:966-969.
Advantages
Saravanan P, Pakshirajan K, Saha P. 2008. Growth Easy online submission
kinetics of an indigenous mixed microbial consortium
Affordable Charges
during phenol degradation in a batch reactor. Biores. Quick processing
Technol., 99:205-209. Extensive indexing
Satsangeee R, Gosh P. 1990. Anaerobic degradation of
phenol using an acclimated mixed culture. Appl. submit@ficuspublishers.com
Microbiol. Biotechnol., 34:127-130.

Haemolymph composition of Ancylostomia stercorea Zeller (Lepidoptera:Pyralidae)

larvae with particular reference to proteins and amino acids
Authors: ABSTRACT:
Labban O, Jugmohan H,
Khan A, Matthew J,
Wisdom S. Haemolymph of 3rd, 4th and 5th larval instars of A. stercorea was extracted and
used in the identification of amino acids by electrophoresis, total protein content by
Lowrys method and proteins by SDS-PAGE. Potassium, sodium and glucose levels
Institution: were also determined. Seventeen amino acids were identified in all three larval
Department of Life Sciences
instars. Thirteen protein bands were present in the haemolymph of 4th and 5th larval
University of the West
Indies. St. Augustine instars, five of which had molecular weights of >45.4 kDa while ten protein bands
Trinidad. West Indies. were present in the haemolymph of 3rd larval instar. Sodium:Potassium ratios in 3rd,
4th and 5th larval instars were 0.05, 0.14 and 0.13 respectively. Glucose concentrations
in 3rd, 4th and 5th larval instars were 0.16, 0.33 and 0.57 mg/ml respectively. The
Corresponding author: results provided critical information that can be used to create artificial diets for in-
Khan A. vitro rearing of A. stercorea parasitoids.
Email: Keywords:
ayub.khan@sta.uwi.edu. Ancylostomia stercorea, haemolymph, larvae, Cajanus cajan.

http://jresearchbiology.com/ Labban O, Jugmohan H, Khan A, Matthew J, Wisdom S.
Documents/RA0199.pdf. Haemolymph composition of ancylostomia stercorea zeller (LepidopteraPyralidae).
Dates:
Received: 16 Feb 2012 /Accepted: 25 Feb 2012 /Published: 13 Mar 2012
Ficus Publishers.
cited.
178-183 | JRB | 2012 | Vol 2 | No 3

Labban et al., 2012
INTRODUCTION to develop artificial diets containing insect haemolymph

Pigeonpea (Cajanus cajan L. Millsp.) is a on which to rear parasitoids, and so should be able to cut
leguminous shrub that originated in India (van der production costs dramatically. Matthews (1974)
Maesen, 2006). It was found in West Africa about concurred that more attention should be paid to the mass
2000BC and the slave trade redistributed it to the West rearing efficiency of parasitoids by inducing species to
Indies, where its use as a bird feed led to the name oviposit directly and to successfully develop upon
pigeonpea in 1692. Pigeonpea forms an essential part artificial nutrient media. However, there are several
of the protein diet found within the Caribbean and can limiting factors hindering the development of these
also be used for a number of purposes including animal artificial diets, including the biochemical interactions
feed and firewood (Shanower et al., 1999). Furthermore, between the insect host and parasitoid and the lack of
pigeonpea improves the soil through its extensive root understanding of entomophage physiology and
system, nitrogen fixation and mulch provided by the metabolism. The objective of this study was to determine
fallen leaves (Van der Maesen, 2006). the haemolymph composition of A. stercorea with
The plant is plagued by numerous insect pests particular reference to protein, amino acid and ion
throughout the Caribbean and other areas where it is composition as a pre-requisite step in artificial rearing of
grown (Khan et al, 2003). They attack all stages and its major parasitoid, Bracon thurberiphagae Muesbeck
parts of the plant from seedling to harvest and beyond. (Hymenoptera:Braconidae).
However, damage caused by lepidopterous pod borers is
very significant since they attack the edible seeds MATERIALS AND METHODS
rendering them unfit for consumption. The most The experiment was carried out during the
important pod borer pest of pigeonpea in the Caribbean months of March May 2010. Pigeonpea pods from
and particularly Trinidad and Tobago is Ancylostomia fields were collected with 3rd, 4th and 5th larval stages of
stercorea Zeller (Lepidoptera:Pyralidae) which causes A. stercorea and kept in cages. Haemolymph from 3rd, 4th
significant damage to green seeds. Buckmire (1979) and 5th instar larvae was extracted by piercing the cuticle
reported that in Trinidad, over 90% of the green pods with a thin, sharp insect mounting pin (No.3) and
could be infested by A. stercorea. Bennett (1960) separately collected in Corning Pyrex 5l disposable
described the pest as a multivoltine species, depositing micropipettes (Corning Science Products, USA).
its eggs on the young pods of pigeonpea; the larvae then Zone electrophoresis of amino acids
feed on the developing seeds and later pupate in soil. The Electrophoresis of amino acids in the
life cycle is completed in 26-32 days. haemolymph of A. stercorea larvae was done using a
Major concerns regarding pesticide overuse and Gelman Semi-micro Electrophoresis chamber with a
its impact on human health and the environment have 180V DC supply. Phosphate buffer (150ml of 0.1M
arisen. A great deal of emphasis has therefore been KH2PO4 was added to 90ml of 0.1M NaOH and diluted
placed on finding alternative solutions (e.g. biological to 600ml giving a pH of 7.02) was poured into the
control) to large scale use of synthetic insecticides. chamber tray until almost half filled. The tray was gently
Guerra et al (1993) notes that mass production of tilted to ensure that the phosphate buffer was equalized
parasitoids for use in biological programmes requires in all compartments. Strips (6 cm x 14 cm) of
very costly in vivo techniques which involve mass Whatman No.1 filter paper were placed on clean paper
production of host insects. Researchers have attempted and a light pencil line was drawn across the centre of
Labban et al., 2012
each strip. One end was marked (+) to be immersed in 10l of TEMED). Ten wells each with 10L /well was
the anode and the other (-) to be immersed in the used for the determination of proteins present in the
cathode. Three equally spaced small dots were marked haemolymph of 3rd, 4th and 5th larval instars with each
on the centre line for application of two standard amino sample comprising: 3L Sample + 3L Sucrose + 3L
acid samples (0.2% (w/v) amino acid solutions preserved Bromophenol Blue + 1L Running Buffer. -
with a drop of toluene) and the other for the haemolymph Lactalbumin from Bovine milk (14.2kDa), Carbonic
sample. 5 l of each sample was transferred to its Anhydrase from Bovine Erythrocytes (29kDa), Albumin
appropriate spot and the strip placed in the chamber and from Chicken egg white (45kDa), and Urease from Jack
ran for 75 minutes at the end of which a glass rod was Bean (545kDa) were used as markers.
used to remove the paper. The paper was then dried with Sodium, Potassium and Glucose determination
a hair dryer for about 3-4 minutes and dipped in 0.3% Sodium and potassium haemol ymph
Ninhydrin solution (0.3% (w/v) solution in acetone + one concentrations were determined using a Sherwood Flame
drop of pyridine added per 50mL of solution) and dried Photometer Model 410 (Sherwood Scientific Ltd, UK).
for a further 4-5 minutes. The appearance of coloured The values presented represent the means of five
spots on the paper indicated the position of each amino replicates per instar. Glucose concentrations for each
acid. Retention factor (Rf) values were then calculated instar were determined using the method of Folin-Wu
from the results collected, using the following formula: (1919).
Rf = Distance moved by spot / Distance moved by
solvent front. These values were compared to standard RESULTS AND DISCUSSION
values of the amino acids used and the amino acids Amino acids
present in the haemolymph samples were identified. Free amino acids, proteins and other organic ions
Protein content and separation are very important components of parasitoid diets (Baker
Total protein concentration in the haemolymph and Fabrick, 2000). Research on the nutritional
was determined using the Lowry et al. (1951) method. components of host larvae which can be utilized by
Bovine Serum Albumin (200 g/mL) was used as the ectoparasitoids to support their larval growth and
standard protein. Proteins were separated using PAGE development is useful for the development of optimal
with a running buffer of pH 8.3 (0.6g Tris, 2.88g glycine artificial diets for the in vitro rearing of these
in 1000ml of de-ionized water) and a protein stain of ectoparasitoids. There were seventeen amino acids
0.2% Coomassie Blue R-250 in 50% methanol : 10% identified in all three larval instars of A. stercorea (Table
acetic acid (0.2g Coomassie Blue R-250, 50ml of 1). Leucine was present in 4th and 5th larval instars but
methanol and 10ml of acetic acid in 100ml of water). absent in 3rd larval instar. Isoleucine was present in 3rd
Gels were de-stained using a solution of 30% methanol: and 5th larval instars and absent in 4th larval instar while
10% acetic acid. A 7.5% separating gel (4.95ml of de- tryptophan was present in 4th and 5th larval instars but
ionized water, 2.50ml of 1.5M Tris-HCl (pH 8.8), 2.50ml absent in 3rd larval instar (Table 1). Chen (1962) noted
of Acrylamide BIS (30% stock), 50l freshly prepared that ten amino acids (arginine, histidine, isoleucine,
10% Ammonium Persulphate and 5l TEMED and a 4% leucine, lysine, methionine, phenylalanine, threonine,
stacking gel (6.2ml de-ionized water, 2.5ml 0.5M Tris- tryptophan, and valine) have been proven to be essential
HCl (pH 6.8), 1.33ml Acrylamide BIS (30% stock), for insect growth and development, all of which were
50l freshly prepared 10% Ammonium Persulphate and present in the insect larvae investigated. Duke (1983)

Labban et al., 2012
Table 1 Amino acids identified by paper significantly different (p<0.05, Tukey-Kramer Multiple
chromatography in haemolymph of Ancylostomia Comparisons Test) between 3rd, 4th and 5th instars of A.
stercorea Zeller (Lepidoptera:Pyralidae) 3rd, 4th and 5th
instar larval stages. stercorea (Table 2) since the same proteins may be
needed for growth and development in each larval stage
Amino acid 3rd Instar 4th Instar 5th Instar
however, they may be present in different concentrations.
Alanine
Indeed changes in the haemolymph biochemistry of
Arginine
Plodia interpunctella after treatment with Bacillus
Asparagine
thuringiensis revealed that there was insignificant
Aspartic acid
variation in the total proteins in normal untreated larvae
Cysteine
(Aboul-Ela et al, 1991). Thirteen protein bands were
Glutamine
present in the haemolymph of 4th and 5th larval instars,
Glutamic acid
five of which had molecular weights >45.4 kDa. Ten
Glycine
protein bands were present in 3rd instar larvae, however
Histidine
most of the bands were found in very low concentrations
Isoleucine X
except for a prolific band with a molecular weight of
Leucine X
18.7 kDa that was also present in the other larval instars.
Lysine Generally high molecular weight proteins were present in
Methonine 4th and 5th instars while proteins of low molecular
Phenylalanine weights were found mainly in 3rd larval instar although
Proline they were also present in the other two larval instars but
Serine in very low concentrations (Plate 1). One probable
Threonine explanation for the numerous protein bands in 4th and 5th
Tryptophan X instar haemolymph may be that most insects require
Tyrosine larger amounts of protein for further growth and
Valine development. Nation (2008) and Kanagalakshmi (2011)
Presence X - Absence confirm that some insects (larvae of Diptera and
Lepidoptera) synthesize large quantities of one or more
revealed the presence of 20 amino acids in Cajanus storage proteins during late larval life and this was
cajan (pigeonpea) namely alanine, arginine, asparagine, particularly evident in the haemolymph of 4th and 5th
aspartic acid, cysteine, glutamine, glutamic acid, glycine, instar A. stercorea larvae (Table 2). Chen (1962) also
histidine, isoleucine, leucine, lysine, methionine, suggested that the total haemolymph protein content
phenylalanine, proline, serine, threonine, tryptophan, increased during larval development and was most rapid
tyrosine and valine. Most of these were found in the during the period approaching pupation. The large
larval haemolymph of A. stercorea investigated number of protein bands present in 4th and 5th instars may
indicating that the larvae of A. stercorea may have be as a result of protein accumulation to sustain growth
derived most of its amino acid content from C.cajan during the pupal stage. Interestingly though, a distinct
seeds. protein band of a molecular weight of 18.7 kDa was very
Protein content and separation prolific in all three larval stages and could possibly be a
Haemolymph protein levels were not major protein needed for growth and development in all
Labban et al., 2012
Table 2 Potassium (K+), sodium (Na+), glucose and total protein content ( SE) in
haemolymph of 3rd, 4th and 5th larval instars of Ancylostomia stercorea (Zeller)
Na+ : K+
Instar Na+ (mg/ml)* K+ (mg/ml)* Glucose (mg/ml)* Total protein (mg/ml)*
Ratio
3rd 0.092 0.003a 1.692 0.002a 0.05 0.16 0.06a 66.0 27.0a
4th 0.153 0.006 b
1.128 0.005 b
0.14 0.33 0.11 ab
85.5 22.5a
5th 0.146 0.002 b
1.128 0.003 b
0.13 0.57 0.09 b
73.5 20.5a
*Values followed by the same letters in a column are not significantly different from
each other based on Tukey- Kramer multiple comparisons test (p< 0.001).
life stages of the insect. groups (Nation 2008). Glucose haemolymph

Potassium, Sodium and Glucose concentrations were significantly different (p<0.05)
Potassium concentrations were not significantly between 3rd and 5th instars (Table 2) and may be
different (p> 0.001) between 4th and 5th instars; however interpreted as the later larval instars requiring larger
rd
3 instar haemolymph had significantly higher (p< quantities of glucose for increased metabolism as the
0.001) potassium levels than the other two instars. A insect approaches pupation (Aboul-Ela et al, 1991;
similar trend was observed for sodium haemolymph Nation 2008; Malik and Malik 2009). The results from
+ +
concentrations (Table 2). Ratios of Na : K are generally the experiment provided critical information that can be
very low ( 0.3 0.1) in Lepidoptera, phytophagous used to strengthen the gaps that exist in creating artificial
Coleoptera, Hemiptera, Homoptera and Hymenoptera diets for in-vitro rearing of parasitoids of A. stercorea.
and may be correlated with the food habits of these
302.6 545
192.3
146.2
110.9
97.1
45.5 29
32.4
26.3
18.7
16.3
11.6 14.2 45
8.2
6.6
Lactalbumin Carbonic Albumin Urease
3rd 4th 5th anhydrase0
Plate 1 Electrophoresis gel of proteins present in haemolymph of 3 rd, 4th and 5th larval
instars of A. stercorea (values are given in kDa, markers used were Lactalbumin,
Carbonic anhydrase, Albumin and Urease)

Labban et al., 2012
REFERENCES reproductive and reproductive molt cycle of Albunea

Aboul-Ela R, Kamel MY, Salama HS, El-Moursy A, symmista. Journal of Research in Biology 1(2): 68-72
Abdel-Razek A. 1991. Changes in the biochemistry of
Khan A, Baker PS, Pollard GV. 2003. Semi-field tests
the haemolymph of Plodia interpunctella after treatment
of the effects of three insecticides on Ancylostomia
with Bacillus thuringiensis. Journal of Islamic Academy
stercorea. Tests of Agrochemicals and Cultivars (Annals
of Science 4(1):29-35.
of Applied Biology, 141, Supplement):23:4-5.
Baker JE, Fabrick JA. 2000. Host haemolymph
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ.
proteins and protein digestion in larval Habrobracon
1951. Protein measurement with the Folin-Phenol
hebetor (Hymenoptera:Braconidae). Insect Biochemistry
reagent. Journal of Biological Chemistry 193:265-275.
and Molecular Biology 30 (10):937-946.
Malik MA, Malik AF. 2009. Ontogenic changes in
Bennett FD. 1960. Parasites of Ancylostomia stercorea
haemolymph biochemical composition in the silkworm,
(Zell.) (Pyralidae:Lepidoptera), a pod borer attacking
Bombyx mori L under thermal stress. Academic Journal
pigeon pea in Trinidad. Bulletin of Entomological
of Entomology 2(1):16-21.
Research 50:737-757.
Matthews RW. 1974. Biology of Braconidae. Annual
Buckmire KU. 1979. Pests of Grain Legumes and their
Review of Entomology 19:15-32.
control in the Commonwealth Caribbean. In Singh SR,
van Emden HF, Taylor TA (Eds) Insect Pests of Grain Nation JL. 2008. Insect Physiology and Biochemistry.
Legumes: Ecology and Control. Academic Press, 3rd Edition CRC Press, USA. 496.
London UK. 454.
Shanower GT, Romus J, Minja ME. 1999. Insect pests
Chen PS. 1962. Amino acid and protein metabolism in of pigeonpea and their management. Annual Review of
insect development. Advances in Insect Physiology 3:53- Entomology 44:77-96.
132.
Van der Maesen LJG. 2006. Cajanus cajan (L.) Millsp.
Duke JA. 1983. Handbook of energy crops. http:// http://database.prota.org/search.htm.
w w w. h or t . p u r d u e . e d u / n e w c r op / d u k e _ e n e r g y/
Cajanus_cajun.html.
Folin O, Wu H. 1919. A system of blood analysis:

Supplement I - A simplified and improved method for
determination of sugar. Journal of Biological Chemistry
Advantages
41:367-374.
Guerra AA, Robacker M, Martinez S. 1993. Free
Affordable Charges
amino acid and protein titers in Anthonomus grandis Quick processing
larvae venomized by Bracon mellitor. Entomophaga 38 Extensive indexing
(4):519-525. You retains your copyright
Kanagalakshmi K. 2011. Fluctuation of protein level in submit@ficuspublishers.com

haemolymph, ovary and hepatopancreas during non-
A comparative study of fish population in Temengor Reservoir and Bersia

Reservoir, Perak, Malaysia
Authors: ABSTRACT:
Muzzalifah Abd Hamid1,
Mashhor Mansor1,
Zarul Hazrin Hashim1, A study on fish diversity and community of two reservoirs in Hulu Gerik was
Mohd. Syaiful carried out from August 2009 to December 2009. The two selected reservoirs were
Mohammad2, Temengor Reservoir and Bersia Reservoir. The aim of this study is to compare the
community structures of freshwater fish population between these two reservoirs. A
Institution:
total of 15 species which comprise of six families were recorded in this study. Twelve
1.School of Biological
Sciences, Universiti Sains species were recorded in Temengor Reservoir whereas 13 species were recorded in
Malaysia, 11800 Minden, Bersia Reservoir. The best represented family in both reservoirs was Cyprinidae with
Penang, Malaysia. eight species in Temengor Reservoir and seven species in Bersia Reservoir.
Cyclocheilichthys apogon was selected for the length-weight relationship and
2. Pulau Banding Rainforest condition factor analysis. C. apogon showed a better growth in Bersia Reservoir in
Research Center, Pulau comparison to Temengor Reservoir.
Banding, 33300 Gerik,
Perak, Malaysia.
Corresponding author:
Muzzalifah Abd Hamid. Keywords:
Fish diversity, reservoir, length-weight relationship, condition factor,
Cyclocheilichthys apogon.
Email:
muzzalifah.abdhamid@gmail.com Article Citation:
Muzzalifah Abd Hamid, Mashhor Mansor, Zarul Hazrin Hashim, Mohd. Syaiful
Mohammad.
Web Address: A comparative study of fish population in Temengor Reservoir and Bersia Reservoir,
http://jresearchbiology.com/
Perak, Malaysia.
Journal of research in Biology (2012)2(3): 184 192
Dates:
Received: 24 Feb 2012 /Accepted: 04 Mar 2012 /Published: 24 Mar 2012
Ficus Publishers.
cited.
184 192 | JRB | 2012 | Vol 2 | No 3

Hamid et al., 2012
INTRODUCTION MATERIALS AND METHODS

Fish is one of the most obvious and crucial Sampling Area
inhabitants of a reservoir ecosystem. Fish are sensitive A monthly study on fish diversity and
indicators of the relative health of aquatic ecosystems community in two reservoirs in Hulu Gerik district,
and their surrounding watersheds (Fausch et al., 1990). Perak, was carried out from August 2009 to December
Their size, community composition and structure often 2009. The two selected reservoirs were Temengor
reflect nutrient status of a water body. In addition, fish Reservoir and Bersia Reservoir (Figure 1). These sites
has a direct effect on other trophic levels, including were chosen due to their location at upper part of Sungai
phytoplankton and zooplankton. Generally, many fish Perak basin which serves as the first two consecutive
species can be considered as top consumers in aquatic man-made lakes. Temengor Reservoir and Bersia
ecosystems (Dallinger et al., 1987). Some fish consume Reservoir are separated by a dam named Temengor Dam
phytoplankton, whereas others consume zooplankton, (completed in 1977). Temengor Reservoir and Bersia
fish or fish larvae. Pollutants that were discharged into Reservoir cover an area of 152km2 and 5.7km2
aquatic environment are likely to accumulate in fish and respectively.
represent a potential risk (Adams et al., 1992). Sampling of Fish
Hashim et al., (2012) recorded that there were 21 Six sets of experimental gill nets (250 cm
species of freshwater fish found in Temengor Reservoir vertical length x 2976 cm total width) with five different
in 2006. This finding indicated some missing species in stretch mesh sizes (3.7 cm, 5 cm, 6.5 cm, 7.5 cm, 10 cm)
comparison with the previous study done by Md. Akhir were soaked randomly overnight in each reservoir. A
(1999) who found 37 species of fish in the same total of 692 individuals were captured in Temengor
reservoir. Unfortunately, to date, there are no Reservoir whereas 214 individuals were captured in
documented studies on fish species featuring Bersia Bersia Reservoir. All captured fish were identified and
Reservoir. Therefore, this study is believed to be the first measured (total length and weight). Species
report on fish population comparison between these two identification was based on taxonomic keys by Mohsin
reservoirs and will thus further contribute to the
development of comprehensive baseline data for
freshwater fishes in these regions.
Fish species have been used as biological
indicators to show the level of environmental quality
towards the threats of aquatic pollution since early
1900s. The concept of using fish communities as
biological indicator has been historically followed by
several authors (Ortmann, 1909; Karr, 1981; Fausch et
al., 1990; Schiemer, 2000). Therefore, this study was
conducted in order to detect changes in two consecutive
reservoirs by using fish community as an indicator. In
addition, this study compares the current status of
Figure 1: Location of Temengor Reservoir and
fisheries between these two reservoirs within the context
Bersia Reservoir at upper part of Sungai Perak
of conservation and management of fisheries. basin (Inset: Map of Peninsular Malaysia)
185 Journal of Research in Biology (2012) 2(3) : 184-192
Hamid et al., 2012
and Ambak (1983) and Rainboth (1996). Reservoir, O.hasseltii and Chela anomalura represented
Shannon Wiener Diversity and Evenness Indices 35% and 20% each of the catches followed by C.apogon.
These indices were applied to fish diversity study However in Bersia Reservoir, C.anomalura was absent
and were computed for each system. The diversity and and Labiobarbus leptochilus and C.apogon represent the
evenness indices were done by using Multi-Variate two highest abundance with 24% and 23% each followed
Statistical Package (MVSP) version 3.13 d. by O.hasseltii (16%) and the other seven species (17%).
Length-Weight Relationship (LWR) A total of four families were recorded in Temengor
In general, the change in weight of the fish can Reservoir while six families were recorded in Bersia
be described by the relationship W=aLb, where Reservoir. The highest abundance of family composition
W=observed fish weight (g) and L=observed fish length is Cyprinidae in both Temengor and Bersia reservoirs.
(cm). Value of a is estimated by using antilogarithm of ln In Temengor Reservoir, Cyprinidae comprised of eight
in the linear regression; ln W= ln a + b ln L, where ln = fish species whereas in Bersia Reservoir, Cyprinidae
regression intercept and b = regression slope. comprised of seven fish species.
Condition Factor (K) Shannon Wiener Diversity and Evenness Indices:
Condition factor refers to the factor of well being Result showed that diversity index in Bersia
and the degree of fatness of fish (Fafioye and Oluajo, Reservoir (2.108) was higher than Temengor Reservoir
2005). Condition factor (K) is presented by the equation (1.783) during the study. Evenness index of fish species
3
K= 100W/L , where W=weight (g) and L=total length in Bersia Reservoir is also higher than Temengor
(cm). Cyclocheilichthys apogon was selected for this Reservoir which represents 0.822 and 0.718,
purpose. respectively. Bersia Reservoir recorded a higher number
Independent-samples t test of species present, but the individuals in the community
Independent-samples t test was computed by were distributed more equally among the species. In
using SPSS version 11.5 for comparing two parameters; Temengor Reservoir, there is one less species compared
total fish catch and total catch of C.apogon between to Bersia Reservoir and most of the individuals belong to
Temengor Reservoir and Bersia Reservoir. O.hasseltii. O.hasseltii is the most common species in
Temengor Reservoir that makes up about 35% of the
RESULT community.
Fish Community Structure Length-Weight Relationship (LWR) of C.apogon
Fifteen species, which comprised of six families, The example form of parabolic equation of
were recorded during this study. Cyprinidae was the C.apogon in Temengor Reservoir and Bersia Reservoir
most dominant family with nine species while the other was shown in Table 2. The estimation for the length-
families were only represented by one or two species weight relationship in Temengor Reservoir is taken by
(Table 1). Twelve species were recorded in Temengor the formula, W=0.004L3.363. From the relationship, the
Reservoir with four families while 13 species were values of a and b are 0.004 and 3.363, respectively. For
recorded in Bersia Reservoir with six families. Total length-weight relationship of C.apogon in Bersia
catch in both reservoirs showed that Osteochilus hasseltii Reservoir, the parabolic equation is represented by
and C.apogon were two species that dominated the W=0.004 L3.414. Based on this formula, the values of a
catches during this study, which represented 30% and and b were 0.004 and 3.414, respectively. The regression
18% of the total catch, respectively. In Temengor coefficient in Bersia Reservoir (R2=0.965) is slightly
Journal of Research in Biology (2012) 2(3): 184 192 186

Hamid et al., 2012
Table 1: Species checklist and total individual catch recorded in

Temengor Reservoir and Bersia Reservoir during this study
Total individual
catch
Temengor Bersia
Bil Family Species
1 Bagridae Mystus nemurus 8 12
2 Bagridae Mystus wyckii - 6
3 Channidae Channa micropeltes 1 9
4 Cyprinidae Labiobarbus leptochilus 31 52
5 Cyprinidae Chela anomalura 141 -
6 Cyprinidae Puntius gonionotus - 5
7 Cyprinidae Puntius schwanenfeldii 1 5
8 Cyprinidae Hampala macrolepidota 88 18
9 Cyprinidae Mystacoleucus marginatus 33 1
10 Cyprinidae Cyclocheilichthys apogon 117 48
11 Cyprinidae Osteochilus hasseltii 241 35
12 Cyprinidae Tor tambroides 1 -
13 Eleotridae Oxyeleotris marmoratus - 1
14 Nandidae Pristolepis fasciatus 19 9
15 Notopteridae Notopterus notopterus 11 13
Total Catch 692 214
Total Species 12 13
higher than Temengor Reservoir (R2=0.920). Figure 2 Bersia Reservoir, the condition factor was 1.3197
and Figure 3 showed the examples of the length-weight 0.118. The value then decrease to 1.0825 0.061 in
relationship of C.apogon in Temengor and Bersia October. Then the value was increased in November
reservoirs, respectively. (1.2664 0.206). A little decrease was then recorded in
Condition factor (K) of C. apogon December (1.2427 0.172). In Temengor Reservoir, the
The condition factor of C.apogon (Figure 4) in condition factor recorded is 1.2090 in August and 1.3227
Temengor Reservoir for October and December were 0.425 in October. The figure shows the decrease in
higher than in Bersia Reservoir. There was no catch of November (1.1592 0.206). A slight increase recorded
C.apogon in Temengor and Bersia reservoirs for in December (1.3181 0.172).
September and August, respectively. For September in
Figure 2: The length-weight relationship of Figure 3: The length-weight relationship of

C. apogon from Temengor Reservoir C. apogon from Bersia Reservoir
187 Journal of Research in Biology (2012) 2(3): 184-192

Hamid et al., 2012
Table 2: Parameter estimates and parabolic equations for C.apogon in Temengor Reservoir and Bersia Reser-
voir in the study. n= total number of samples, min=minimum value, max= maximum value, a= intercept of
regression line, b= slope of regression line, R2 = regression coefficient.
Total Length Weight (g)
(cm)
Reservoir n Min Max Min Max a b R2 W = aLb
Temengor 117 10.2 23.3 10.0 155.0 0.004 3.363 0.920 W = 0.004 L3.363
Bersia 48 11.0 21.7 12.2 123.6 0.004 3.414 0.965 W = 0.004 L3.414
Independent-samples t test Gangapur Dam, India as well.

Independent-sample t test was used to test if GueGan et al., (1998) suggested that the
Bersia Reservoir was affected by Temengor Dam. Based components of reservoir size (surface area) and energy
on the Table 3, the two parameters were not differed availability (net primary productivity) are the most
significantly (p>0.05) between the Temengor and Bersia important factors in predicting fish diversity.
reservoirs. This means that total individual fish catch and Nevertheless, the role of other possible factors such as
condition factor of C.apogon at Bersia Reservoir were current climate and history of the reservoir are merely
relatively similar to Temengor Reservoir. marginal importance. The difference between species
diversity in Temengor Reservoir and Bersia Reservoir
DISCUSSION may due to the environmental changes especially logging
A total of 906 individual fish comprised of six activities. According to Samat et al., (2002) logging
families were recorded in both Temengor and Bersia activities will change the water body from being shaded
reservoirs. However, when compared to previous studies, with canopy cover to exposed water body with little
the declining trend in species diversity in Temengor cover. Logging activity also plays a major role in water
Reservoir was noted. A total of 37 species with 15 body through the changes in hydrological regimes, water
families were recorded in Temengor Reservoir by Md quality, amounts of organic debris, sediment transport
Akhir (1999) in comparison to this study which recorded and quantity, and primary and secondary production
only 12 species with five families. (Campbell and Doeg, 1989).
The highest percentage of family composition in
both Temengor and Bersia reservoirs is Cyprinidae.
Zakaria-Ismail (1996) reported that the Cyprinidae was
the most common family in Malaysian freshwater
bodies. This largest family of all freshwater fishes
occupies virtually all habitats throughout their
distribution (Howes, 1991; Nelson, 1994). This family is
highly adapted in any body forms and mouth structures
(Ward-Campbell et al., 2005). Taki (1978) highlighted
that in Southeast Asia, the distributional summit of
cyprinids may contribute to 40% or more of the species
in a watershed. In addition, Rathod and Khedkar (2011) Figure 4: Condition factor (mean SD) of C. apogon
noted that Cyprinidae was the dominant family in in Temengor Reservoir and Bersia Reservoir
Journal of Research in Biology (2012) 2(3): 184-192 188

Hamid et al., 2012
the parameters of the length-weight relationship. Based
Table 3: Independent-sample t test
on the length-weight relationship of C.apogon in both
Parameter p Mean
reservoirs, every individual fish can influence the
Total individual
0.143, p>0.05 0.9563 0.5786 accuracy of regression coefficient (R2). The length or
fish catch
Condition factor other physical dimension including weight can be
0.721, p>0.05 0.0244 0.0653
of C.apogon influenced by environmental factor (Shukor et al., 2008).
The fish community and distribution can be very Shamekhi et al., (2012) reported that the inter-basin
diverse, and it depends on both biotic and abiotic differences in length and weight of the fish were
conditions of reservoir such as food availability, size of influenced by the availability of food resources and fish
the water body, oxygen content and the primary growth.
production of plants and algae (Gophen et al., 1998). The coefficient of condition is also used to
Factors such as increased fishing pressure, open access determine the suitability of the environment for
and unregulated fisheries, and biologically incompatible particular species. The change of length-weight
water level management of reservoir can affect the relationship of fish is influenced by environmental
fisheries of reservoir, negatively (Petr, 1995). factors. If the environmental factor favours the fishs
Damming plays a significant role as external growth, the fish will show b value that close to 3.0
factor. The formation of reservoir initially boosts food (Yousof et al., 1989). Results have shown that C. apogon
availability and presents high water level variations in Bersia Reservoir has higher b value than Temengor
(Agostinho et al., 1999). Ambak and Jalal (1998) Reservoir. This indicated that C. apogon has better
documented the changes occurred in fish fauna and the growth in Bersia Reservoir as compared to Temengor
patterns of habitat use as the result of the inundation of Reservoir. However, the b value that was closer to 3 in
the man-made Lake Kenyir. Moreover, factors such as Temengor Reservoir showed that C apogon in Temengor
water transparency, water velocity in fish habitats, and Reservoir has the ideal shape of fish growth.
food availability should be considered to analyze the The growths of fish depend on the availability of
effects on fish composition. food and habitat. Usually, fish that have adequate food
During this study, the gill nets were placed and suitable habitat occupation are heavier than normal
randomly at the tributary. Generally, tributaries were weight at a certain length (Jobling, 2002). Regarding to
differed from the main water body with respect to the environmental factors that influence formation of
environmental properties and the local heterogeneity growth pattern in tropical fishes, Santos and Barbieri
(Benda et al., 2004). The fish density and community (1993) highlighted that it can diverge in intensity within
were higher in tributaries compared to main water body and between years.
(Brown and Coon, 1994) because tributaries have Condition factors have been widely used to get
become main habitats for early life stages of fish (Rice et the information about biological status of fish. The term
al., 2001). condition can be used to express the fishs general well
The length-weight relationship parameters allow being (Le Cren, 1951). Condition factor of C.apogon was
calculation and estimation of fish condition. Among the measured based on the number of total catch of the
fish from the same length, the heaviest will be in better species, in both Temengor and Bersia reservoirs. The
condition (Bagenal and Tesch 1978). In this study, a higher K value of C. apogon in Bersia Reservoir
2
square of regression coefficient (R ) was used to estimate indicated that this reservoir provided a more favourable

Hamid et al., 2012
environment for this species as compared to Temengor in both Temengor and Bersia reservoirs is Cyprinidae.
Reservoir. The higher K value of C. apogon in Bersia Reservoir
The difference of K values are influenced by age indicated that this reservoir provided a more favourable
of fish, sex, season, stage of maturation, fullness of gut, environment for this species as compared to Temengor
type of food consumed, amount of fat reserve and degree Reservoir. Results showed that the fish population found
of muscular development (Ozaydin et al., 2007; Cherif et in Temengor Reservoir was relatively similar with Bersia
al., 2008). In some fish species, the gonads may weigh Resevoir. Therefore, Temengor Dam has no effect on
up to 15% or more of total body weight. The K value will fisheries in Bersia Reservoir.
decrease rapidly in females when the eggs are shed
(Barnham and Baxter, 1998). The K value is greatly ACKNOWLEDGEMENT
influenced by the development of stages in reproductive We would like to thank Universiti Sains
organs. During the study, there is a month that the Malaysia and Pulau Banding Rainforest Research Centre
C.apogon is spawning and some of them are in different for providing physical facilities in order to carry out this
stage of the reproductive cycle. study. Funding for this project was provided by Research
Generally, C.apogon moves to flooded forest and Grant 1001/PBIOLOGI/815038.
on forested floodplain. This species was found in the
impoundment and seems to prosper here. As noted by REFERENCES
Rainboth (1996), this species was known to breed late in Adams WJ, Kimerle KA and Barnett JJW. 1992.
the high-water season from September to October as Sediment Quality and Aquatic Life Assessment.
water levels peak and begin to decline. This movement is Environmental Science and Technology 26:1865-1875.
basically to avoid predation by other species that move
Agostinho AA, Miranda L, Bini L, Gomes L, Thomaz
back to the rivers immediately at the onset of falling
S and Suzuki H. 1999. Patterns of Colonization in
water. Moreover, this harmless species is rarely used in
Neotropical Reservoirs and Prognoses on Aging. In:
aquaculture or as bait (Chheng et al., 2005).
Tundisi, J., Straskraba, M. (Eds.). Theorical Resevoir
The presence of dam in consecutive reservoirs
Ecology and its Applications. 227-265.
will influence the environmental conditions in the latter
reservoir. This is because the dam led to significant Ambak MA and Jalal KCA. 1998. Management and
changes in the rivers annual hydrographic due to the Ecological Note Habitat Utilization by the Tropical Fish
large storage reservoir it impounds (McAdam et al., Community in the Man-Made Lake Kenyir, Malaysia
1999). However, the two parameters tested in this study Fisheries Management and Ecology 5:173-176.
which were the total of fish catch and the condition
Bagenal TB and Tesch FW. 1978. Methods for
factor of C.apogon in Bersia Reservoir were relatively
Assessment of Fish Production in Fresh Waters, 3 rd
similar with Temengor Reservoir. Therefore, Temengor
Edition, Blackwell Scientific Publications, Oxford. Age
Dam has no effect on fisheries in Bersia Reservoir.
and Growth. pp. 101136. In:T. Bagenal (ed.)
CONCLUSION Barnham C and Baxter A. 1998. Condition Factor, K,

Twelve fish species were recorded in Temengor for Salmonid Fish. Fisheries. 1-3.
Reservoir whereas 13 species were recorded in Bersia
Reservoir. The highest percentage of family composition

Hamid et al., 2012
Benda L, Poff NL, Miller D, Dunne T, Reeves G, Pess GueGan JF, Lek S and Oberdorff T. 1998. Energy
G and Pollock M. 2004. The Network Dynamics Availability and Habitat Heterogeneity Predict Global
Hypothesis: How Channel Networks Structure Riverine Riverine Fish Diversity. Nature 39:382-384.
Habitats. Bioscience 54:413-427.
Hashim, ZH, Shah ASRM, Mohammad MS, Mansor
Brown DJ and Coon TG. 1994. Abundance and M and Sah SAM. 2012. Fishes of Sungai Enam and
Assemblage Structure of Fish Larvae in the Lower Sungai Telang in Temengor Reservoir, Perak, Malaysia.
Missouri River and Its Tributaries. Transactions of the Checklist, 8(1): 027-031.
American Fisheries Society, 123:718-732.
Howes GJ. 1991. Systematic and Biogeography: An
Campbell IC and Doeg TJ. 1989. Impacts of Timber Overview. In: Winfield, I.J., Nelson, J.S. (Eds), Cyprinid
Harvesting and Production on Streams: A Review. Aust. Fishes: Systematic, Biology and Exploitation. Chapman
J. Mar. Freshwater Res., 40:519-539. & Hall, London, 1-33.
Cherif M, Zarrad R, H. Gharbi H, Missaoui and Jobling M. 2002. Environmental Factors and Rates of
Jarboui O. 2008. Length-Weight Relationships for 11 Development and Growth. Hanbook of Fish Biology and
Fish Species from the Gulf of Tunis (SW Mediterranean Fisheries. Blackwell Science Ltd. 97-117.
Sea, Tunisia). Pan-American Journal of Aquatic
Karr JR. 1981. Assessment of Biotic Integrity Using
Sciences, 3(1): 1-5.
Fish Communities. Fisheries, 6(6): 21- 27.
Chheng, P, Thang, TB, Baran E and Vann LS. 2005.
Le Cren ED. 1951. The Length Weight Relationship and
Biological Reviews of Important Cambodian Fish
Seasonal Cycle in Gonad Weight and Condition in Perch
Species, based on Fishbase 2004, WorldFish Center and
(Perca fluviatilis).J.Animal Ecol., 20:12-16.
Inland Fisheries Research and Development Institute,
Phnom Penh, Cambodia, 1: 66-73. Mcadam DSO, Liley NR and Tan ESP. 1999.
Comparison of Reproductive Indicators and Analysis of
Dallinger R, Prosi F, Segner H and Back H. 1987.
the Reproductive Seasonality of the Tinfoil Barb,
Contaminated Food and Uptake of Heavy Metals by
Puntius Schwanenfeldii, In the Perak River, Malaysia.
Fish. Oecologia 73:91-98.
Environmental Biology of Fishes 55:369-380.
Fafioye OO and Oluajo OA. 2005. Length- Weight
Md. Akhir MZ. 1999. Habitat Heterogeneity and Fish
Relationship of Five Fish Species in Epe Lagoon,
Diversity in Temengor River Basin. Bsc. Thesis. Science
Nigeria. African J. of Biotechnology 4(7):749-751.
University Of Malaysia, Penang. 74.
Fausch KD, Lyons J, Karr JR and Angermeier PL.
Mohsin AKM and Ambak MA. 1983. Freshwater
1990. Fish Communities as Indicators of Environmental
Fishes of Peninsular Malaysia. Penerbit Universiti
Degradation. American Fisheries Society Symposium
Pertanian Malaysia, Serdang. 284.
8:123-144.
Nelson JS. 1994. Fishes of the World. John Wiley &
Gophen M, Yehuda Y, Malinkov A and Degani G.
Sons Inc., New York, New York, USA. 600.
1998. Food Composition of the Fish Community in Lake
Agmon. Hydrobiologia, 380:49-57. Ortmann AE. 1909. The Destruction of the Fresh-
Water Fauna in Western Pennsylvania. Proceedings of

Hamid et al., 2012
the American Philosophical Society, 48(191): 90-110. Shukor MN, Samat A, Ahmad AK and Ruziaton J.
2008. Comparative Analysis of Length-Weight
Ozaydin O, Uckun D, Akalin S, Leblebici S and
Relationship of Rasbora Sumatrana In Relation To the
Tosunoglu Z. 2007. Lengthweight Relationships of
Physiochemical Characteristic in Different Geographical
Fishes Captured from Izmir Bay, Central Aegean Sea. J.
Areas in Peninsular Malaysia. Malays. Appl Biol., 37
Appl. Ichthyol, 23: 695696.
(1):21-29.
Petr T. 1995. The Present Status Of and Constraints to
Taki Y. 1978. An Analytical Study of the Fish Fauna of
Inland Fishery Development in Southeast Asia. In: Petr,
the Mekong Basin as a Biological Production System in
T. and Morris, M. Ed. Indo-Pacific Fishery Commission,
Nature. Research Institute for Evolutionary Biology,
FAO Report: 512- 529.
Tokyo Special Publication 1:77.
Rainboth WJ. 1996. Fishes of the Cambodian Mekong.
Ward-Campbell BMS, Beamish FWH and
FAO Species Identification Field Guide for Fishery
Kongchaiya C. 2005. Morphological Characteristics In
Purposes. FAO, Rome, 265.
Relation To Diet in Five Co-Existing Thai Fish Species.
Rathod, SR and Khedkar GD. 2011. Impact of Journal Fish Biology 67:1266-1279.
Elevation, Latitude and Longitude on Fish Diversity in
Yousof AR, Firdous G, Balkhi MH and Anil AP.
Godavari River. Journal of Research in Biology, 4:269-
1989. Studies on the Length-weight Relationship in
275.
Some Cyprinid Fish in Manasbal Lake, Kashmir. Current
Rice SP, Greenwood MT and Joyce CB. 2001. Trends in Fish and Fishery Biology and Aquatic Ecology
Tributaries, Sediment Sources, and the Longitudinal 185-188.
Organisation Of Macroinvertebrate Fauna Along River
Zakaria-Ismail M. 1996. Freshwater Fish Biodiversity.
Systems. Canadian Journal of Fisheries and Aquatic
Paper presented at the CAP-SAM National Conference,
Sciences 58:824-840.
State of Malaysian Environment. Consumers
Samat A, Shukor MN and Mazlan AG. 2002. Association of Penang. 5 9 January, Penang, Malaysia,
Diversity and Conservation Status of Fishes Inhabits pp: 233-237.
Malaysian Highlands Stream Systems. Journal of
Wildlife and Parks 20:109-118.
Santos G and Barbieri G. 1993. Idade E Crecimento

Do Piau Gordura, Leporinus Piau Fowler, 1941, Na Submit your articles online at Ficuspublishers.com
Represa De TrEmarias (Estado De Minas Gerais)
Advantages
(Pisces, Ostariophysi, Anostomidae). Rev Brasil. Biol., Easy online submission
53(4):649-658. Complete Peer review
Affordable Charges
Quick processing
Shamekhi KR, Patimar R, Ghorbani R and Kordjazi
Extensive indexing
Z. 2012. Comparison Relative of Abundance of Capoeta Open Access and Quick spreading
capoeta gracilis in Five Streams of Gorganroud River You retains your copyright
Basin, Golestan Province, Northern Iran. Journal of submit@ficuspublishers.com
Research in Biology, 1: 019-022.

Assessment of Air Quality through Biomonitors of selected sites of

Dindigul town by air pollution tolerance index approach
Authors: ABSTRACT:
Sarala Thambavani D1 Rapid industrialization has led to different facts of pollution. Vegetation can
and Prathipa V2. absorb particulate and other gaseous pollutants into their system. But they also have
some limitation and tend to show symptoms of damages after prolonged exposure.
Based on the absorbing power and tolerance limit, vegetations can be classified as
highly tolerant, moderately tolerant and sensitive. This has been incorporated by
scientists into a quantitative value of Air pollution Tolerance index (APTI) depending
Institution:
on the score of the plant physiology indicators viz., leaf extract pH, Relative Water
1. Research and
Development Centre Content (RWC), Ascorbic acid and Chlorophyll content. The vegetation monitoring in
Bharathiar University, terms of its APTI act as Bioindicator of air pollution and can be incorporated into
Coimbatore Sri Meenakshi assessment studies. The present investigation were undertaken to assess the air
Government Arts College for quality of selected sites of Dindigul Town. A total of six plant species were collected at
Women (autonomous), three different spots. The results were used to calculate the APTI (Air Pollution
Madurai, Tamil Nadu. Tolerance Index) for each plant and their tolerance/sensitivity were assessed. In this
study, in industrial area Thyme rosemary showed the highest APTI followed by
2. PSNA college of Moringa tinctoria, Calotropis gigantea, Delonix regia, Azadiracta indica and Cynodon
engineering and technology, dactylon. In traffic area Delonix regia, showed the highest APTI followed by
Dindigul, Tamil Nadu. Azadiracta indica, Moringa tinctoria, Calotropis gigantea, Thyme rosemary and
Cynodon dactylon. In Residential area, Cynadon dactylon showed the highest APTI
followed by Calotropis gigantea, Delonix regia, Moringa tinctoria, Thyme rosemary
and Azadiracta indica, Therefore, these plant species could act as the bio indicators
for pollutants and could be utilized as tolerance species towards combating air
pollution.
Corresponding author: Keywords:

Sarala Thambavani D. Biomonitoring, APTI, Bioindicator, Sensitive Tolerant, Ascorbic acid,
Chlorophyll.

http://jresearchbiology.com/ Sarala Thambavani D and Prathipa V.
Documents/RA0205.pdf. Assessment of Air Quality through Biomonitors of Selected sites of Dindigul Town by
Air Pollution Tolerance Index Approach.
Dates:
Received: 29 Feb 2012 /Accepted: 09 Mar 2012 /Published: 04 Apr 2012
Ficus Publishers.
cited.
193-199 | JRB | 2012 | Vol 2 | No 3

Thambavani and Prathipa, 2012
INTRODUCTION bioindicators of pollution. Impact of plant community

Dindigul is a fast developing industrial and has also been studied worldwide in terms of plant-
commercial town and day by day increasing its environment interactions, since the plants are much more
geographical size. Air pollution is caused mainly due to sensitive in comparison to other organisms.
industrial emission, motor vehicle emissions, domestic The symptoms or effect including changes in the
and construction activities. The initial step towards plant anatomy, physiology or biochemistry indicate a
improving the air pollution situation is the monitoring of polluted environment. Thus, the regular monitoring of
the air quality. certain parameters of the plant physiology can indicate
Todays growing population and increasing air pollution in terms of its severerity and degree.
urbanization has resulted in deterioration of ambient air Although sensitivity towards air pollution varies
quality. Air pollution is causing vast changes in across the plant community and some being tolerant
vegetation. Since plants are stationary and they are showing no or minimal symptoms even if the air
continuously exposed to chemical pollutants from the pollutants increase in small amounts the impact can be
surrounding atmosphere, air pollution injury to plants is used for monitoring of air pollution in medium to large
proportional to the intensity of the pollution .Reduction towns and cities in terms of air pollutant concentration to
in plant height, canopy area, plant biomass chlorophyll, observe the air quality of the locality. The plant response
ascorbic acid and nitrogen content in plant growing at to air pollution varies from species to species and also in
sites receiving higher pollution are some of the common terms of type of pollutant, its reacting mechanism,
responses as mentioned by Pandey and Agarwal concentration and duration of exposure Sarala and
(1992),Sarala et al.,(2012) and Chattopadhyay (1996) Sabitha(2012). The pollutants enter into the plants and
reported that leaves respond to pollution and undergo react in variety of ways before being removed or
quantitative change in varying degree in a number of leaf absorbed that may include accumulation, chemical
surface micro morphological characters. transformation and incorporation into the metabolic
The leaves are generally used as experimental system. In this process, some plants are injured while
material as they take up large amount of pollution others show minimal effects (Choudhury et al, 2009).
(Treshow, 1985). APTI is a species dependent plant The trees such as Azadiracta indica, Delonix regia are
attribute which expresses the inherent ability of the plant found to be very common in the urban area of Dindigul.
to encounter stress emanating from pollution. According Near the tannery area the shrub and herbs such as
to Mashita and Pise, (2001) there is a scale of APTI Moringa tinctoria, Calotropis gigantea, Thyme rosemary
value which indicates that APTI value between 30 100 and Cynodon dactylon are found in greater extent .These
shows that the species is tolerant; APTI value between six plant species have been chosen for the Analysis of
17-29 are intermittently tolerant; and plants registering bio chemical parameters. In order to find out the
APTI value in the range of 1-16 are considered as variation in tolerance of pollution on Trees, Shrubs and
sensitive; APTI value lower than 1 is branded as highly Herbs have been taken for analytical study. Keeping in
sensitive. According to Tiwari and Bansal (1993) APTI view of the importance of APTI value in formulating
level of 25 species were found to be different at Bhopal phyto-optimization of air quality, biomonitoring of air
and plants having higher APTI value are more tolerant to quality of selected sites of Dindigul Town were under
air pollution than those having lower APTI value. taken. Tree species which are evergreen such as
Species having lower APTI value may act as Azadiracta indica and, Delonix regia, Shrubs such as
Moringa tinctoria, Calotropis gigantea and Herbs like

Thyme rosemary and Cynodon dactylon were selected to
find the impact of air pollution on biochemical Where A = Ascorbic acid content (mg/g); T =
parameters. In order to compare the effect of air pollutant Total chlorophyll content of leaf (mg/g/fw); P = pH of
on ever green trees, shrubs and herbs these plant species the leaf extract and R = Relative water content of leaf
have been chosen for the present research work. The extract (%)
important findings are reported in this communication. Based on the development and evaluation of
APTI values among the samples they were categorized
MATERIALS AND METHODS into three groups, namely <10 sensitive, >10 <16 is
A total of six different plant species were intermediate species and >17 is tolerant species.
selected for the study from three different spots from
Dindigul Town. The study sites include Residential area RESULTS AND DISCUSSION
(lakshmanapuram), Traffic cum commercial area (Bus The analyzed value for all the six plant species in
stand) and industrial area (Tannery). The screening and three different selected spots of Dindigul Town has been
selection of the plant species are partly based on presented in Table 1. From the results it was found that
literature survey of similar work and guidelines of the pH of the leaf extract was found to be acidic in nature
Central Pollution Control Board (CPCB-1999). in case of almost of all the plant species in all the
Composite leaf samples were sampled in polyethelene sampling sites. But a variation exists in Delonix regia at
bags, tagged brought to the laboratory and analyzed for all the three sites and in Calotropis gigantea at the
several biochemical parameters. At each sampling site residential and tannery area. The acidic nature
composite leaf sample were collected in replications. demonstrates that the air pollutants are mostly gaseous
Now on this pattern of work, a total of six plant species types namely SO2, NOx , diffuse and form acid radicals
were selected for sampling from the above mentioned in leaf matrix by reacting with cellular water. This
area. Again to trace the seasonal variation in the APTI further affects the chlorophyll molecules. (Singh,1991).
value the sampling was done twice, once in winter Among the three sampling sites, the pH of the leaf
season and the other in summer season. The Air extract of Moringa tinctoria and Calotropis gigantea at
pollution Tolerance Index (APTI) was developed by the tannery area are found to be 7.20 and 8.52
analyzing the biochemical parameters of leaf materials, respectively.
viz., pH, ascorbic acid, relative water content and total Relative water content is a useful indicator of the
chlorophyll. ( Pandey and Sharma, 2003). The pH state of water balance of a plant. The large quantity of
values were estimated by using a digital calibrated pH water (in terms of RWC) in plant body helps in
meter. Ascorbic acid, total chlorophyll of leaf extract maintaining its physiological balance under stress
was estimated by spectrophotometric method. Relative conditions of air pollution (Gonzalez and Gonzalez-
water content was estimated by gravimetric method by Vilar, 2001). The Relative Water Content (RWC) of
determining the leaf weight under different condition like leaves is an indicator of the plants water status with
initial, turgid and dry weight. The formula used to respect to its physiological consequences of cellular
determine the Air Pollution Tolerance Index (APTI) water. In residential area it ranged between 62.8% to
using the four parameter is given by (Singh,1991) 75.6% among the studied plant species. In traffic area it
varied between 55.3% to 65.0% for all the plant species.

Table 1 Biochemical constituents in leaf samples collected from six different

plant species at three different spots of Dindigul Town.
Sampling Station
Name of Sampling Station (Traffic) Sampling Station (Tannery)
(Residential)
the plant
Species Total Total Total
pH RWC AA pH RWC AA pH RWC AA
Chl Chl Chl
Azadiracta
5.85 62.8 0.85 2.08 5.55 65 0.478 4.052 5.78 75 0.69 2.62
indica
Delonix
7.53 65.3 0.531 2.65 9.65 55.3 0.384 3.58 6.92 79 0.76 3.15
regia
Moringa
6.75 72.05 0.948 1.65 5.5 62.5 0.636 3.55 7.2 82 0.48 3.68
tinctoria
Calotropis
7.6 70.5 0.674 2.56 4.56 58.5 0.47 3.65 86.5 0.43 1.89
gigantea 8.52
Thyme
5.24 72.3 0.8 1.96 4.85 62.5 0.458 2.35 5.2 72.8 0.56 1.08
rosemary
Cynodon
6.35 75.6 0.66 3.88 3.65 56.5 0.42 4.56 6.6 77.5 0.37 2.34
dactylon
Units as expressed as mg/g fresh weight except pH and RWC content
which is expressed in units and percentage respect.
But in tannery area it ranged between 72.8% to 86.5% main organ of trapping sunlight and its conversion to
for all the plant species. The highest value of RWC was chemical energy. The total chlorophyll content of
recorded by Calotropis gigantea in tannery area Azadiracta indica at the residential, traffic and industrial
whereas the lowest value of RWC was recorded by area are 0.85 mg/g, 0.475 mg/g and 0.69 mg/g
Delonix regia in traffic area of Dindigul Town. The respectively. It showed 43.76% reduction at the traffic
Relative water content of Azadiracta indica is less at area and 18.82% reduction at tannery area. The total
different sampling sites compared to herb Cynodon chlorophyll content for all the sampling sites for Delonix
dactylonbecause the Azadiracta indica is a tall tree regia are 0.531 mg/g, 0.384 mg/g and 0.76 mg/g
which reaches the height of 15-20m and the branches are respectively. The total chlorophyll content of Azadiracta
wide spread and so the evaporation of water is more .But indica at the three different sampling sites such as
the herbs which is very short and growing to 2-15cm Residential, Traffic and Tannery sites are found to be
height and the evaporation of water is less .Hence it has more compared to the Delonix regia. The arrangement of
more relative water content compared to Azadiracta leaves in Azadiracta indica is different from Delonix
indica. regia.It reason out the variation in photosynthetic
The relative water content indicates change in pigment in the tree species.
leaf matrix hydration condition and will generate higher Total chlorophyll content of Moringa tinctoria
acidity condition when RWC is low. More water will at the three sampling sites were 0.948 mg/g , 0.636 mg/g
dilute acidity. From the results it can also be concluded and 0.48 mg/g respectively. It showed 32.91% and
that the former species tends to be more tolerate to air 49.36% reduction at the traffic and tannery area
pollution stress while the later is sensitive. respectively. Calotropis gigantea recorded the total
Total chlorophyll content comprises of chlorophyll content at the three sampling sites at 0.674
Chlorophyll a, Chlorophyll b and other accessory mg/g , 0.47 mg/g and 0.43 mg/g respectively. It showed
pigment. It provides greenness to the leaves and is the 30.26% and 36.20% reduction at the traffic and tannery
area. Thyme rosemary recorded the total Chlorophyll The calculated APTI value associated with the
content as 0.80 mg/g, 0.458 mg/g and 0.56 mg/g sampled vegetation at the residential area ranges between
respectively. It also showed 42.75% and 30% reduction 7.67 to 10.27, 7.49 to 9.08 at the traffic area and 9.3 to
traffic and tannery area. Similarly Cynodon dactylon 13.5 at the tannery area are shown in Table2. All the six
showed the reduction of total chlorophyll at the traffic plant species at Dindigul Town, both at the control and
and tannery area as 36.36% and 43.93% respectively. the polluted area showing APTI value < 17 representing
The higher level of total chlorophyll content in the leaves sensitive response to air pollution. APTI for Azadiracta
of all the six selected species at residential area in indica at the Residential, Traffic and Tannery area are
comparison to traffic and tannery area indicate lower air 7.673, 8.941 and 9.915 respectively. Due to the air
pollution stress in residential area. The lowest value of pollution, the sensitive species moves towards tolerance.
total chlorophyll content in the leaves of six selected The APTI for Delonix regia at the different sampling
species at traffic and tannery area in comparison to sites are 8.66, 9.08and 10.31 respectively. At the tannery
residential area indicates higher air pollution stress. site, it changed in to intermediate tolerance. Moringa
Ascorbic acid, a stress reducing factor is a strong tintoria and Calotropis gigantea have the APTI values
reducing agent and is associated with tolerant plants. It as 8.475, 8.428, 11.02, 9.168, 7.68 and 10.34
reduces the effect of SO2 and acts as antioxidant. A high respectively. Due to the air pollutant from traffic and
content of ascorbic acid in plant leaf is related to tannery sites, the tolerance indexes have been changed.
biochemical and physiological species of a particular Thyme rosemary has the APTI value as 8.1, 7.49 and
environment. It provides specific physiological defense 13.5 at the different sampling sites. Similar APTI values
mechanism as for plants internally and its reducing were recorded for Cynodon dactylon. APTI
power is directly proportional to its concentration categorization of selected plant species of Dindigul
(Khattab, 2007) and Sarala et al.,(2011). In the present Town at different sampling sites were tabulated in
study at the residential area it ranged between 1.65 mg/g Table 2a.
to 3.88 mg/g with Cynodon dactylon having highest and This species having APTI less than 17 APTI
Moringa tinctoria having lowest content. For all the six value can be utilized as bio-indicator of the air quality
plant species at the traffic area the ascorbic acid content in the study area.
ranged between 2.35 mg/g to 4.56 mg/g with Thyme The correlation matrix given in the Table 3, 4, 5
rosemary recording the lowest value and Cynodon signified the association of the four biochemical
dactylon showing the highest value. In tannery area the parameters among themselves and also with the
Ascorbic acid content ranged between 1.08 mg/g to 3.68
Table 2 Comparative APTI values for plant species
mg/g with Thyme rosemary showing lowest and
at selected sites of Dindigul Town.
Moringa tinctoria having highest content. Ascorbic acid APTI values for plant species at
of six test species in traffic and tannery road side plants Plant species different sampling sites
Residential Traffic Tannery
(polluted plants) was increased. The increase in ascorbic
Azadiracta indica 7.673 8.941 9.195
acid might act as strong reductant for defense Delonix regia 8.66 9.08 10.31
mechanisms against automobile pollutants. It is evident Moringa tinctoria 8.475 8.428 11.02
that ascorbic acid activates many physiological and Calotropis
9.168 7.68 10.34
gigantea
defensive mechanisms. Its reducing power is directly
Thyme rosemary 8.41 7.49 13.5
proportional to its concentration. Cynodon dactylon 10.27 7.5 9.38

Table 2a APTI categorization of selected plant showed positive correlation with pH, RWC and Ascorbic
species of Dindigul Town acid but negative correlation with Total chlorophyll.
Categories of Plant species APTI showed positive correlation with RWC and
Plant Residential Traffic Tannery Ascorbic acid. In traffic area APTI showed positive
species
Azadiracta Sensitive Sensitive Sensitive correlation with all the four biochemical parameters.
indica Air Pollution Tolerate Index (APTI) is an
Delonix Sensitive Sensitive Intermediate
regia inherent quality of plants to encounter air pollution
Moringa Sensitive Sensitive Intermediate stress, which is presently of my concern particularly of
tintoria
Calotropis Sensitive Sensitive Intermediate urban areas in the world.
gigantea
Thyme Sensitive Sensitive Intermediate
Rosemary CONCLUSION
The present study suggests that plants have the
Cynodon Interme Sensitive Sensitive
dactylon diate potential to serve as excellent Morphometric,
Quantitative and Qualitative indices of pollution level.
dependent parameter APTI. It was found that there is a Biomonitoring of plants is an important tool to evaluate
positive correlation between APTI and pH, RWC, total the impact of air pollution on plants. Based on the
chlorophyll and ascorbic acid. In residential area APTI results it can be said that the Air Pollution Tolerance
Table 3 Correlation between the APTI values and biochemical parameters estimated from
the leaf samples of the selected spots of Residential area
Total
pH RWC Ascorbic acid APTI
chlorophyll
pH 1.00
RWC -0.124 1.00
Total
-0.530 0.077 1.00
Chlorophyll
Ascorbic acid 0.208 0.373 -0.674 1.00
APTI 0.291 0.743 -0.496 0.863 1.00
the leaf samples of the selected spots of Traffic cum Commercial area
Total
pH RWC Ascorbic acid APTI
Chlorophyll
pH 1.00
RWC -0.324 1.00
Total
-0.286 0.601 1.00
Chlorophyll
Ascorbic acid -0.142 -0.307 -0.115 1.00
APTI 0.783 0.114 0.038 0.187 1.00
the leaf samples of the selected spots of Tannery area
pH RWC Total Chlorophyll Ascorbic acid APTI
pH 1.00
RWC 0.989 1.00
Total Chlorophyll -0.378 -0.391 1.00
Ascorbic acid 0.311 0.330 0.226 1.00
APTI -0.360 -0.290 -0.006 -0.504 1.00
Index (APTI) values estimated using the four Pandey J and Agarwal M. 1992. Ozone concentration
biochemical parameters in plant leaves viz., Relative variabilities in a seasonally dry tropical climate. Environ
Water Content, total chlorophyll, pH and ascorbic acid International 18:515-520.
can be used as predictor of air quality. These parameters
Pandey J and Sharma MS. 2003. Environmental
are significant in studies in plant environment
Science Practical and Field. Yash Pub., Bikaner, India
interactions and used for development of bioindicator
129.
groups. The APTI of particular geographical area can be
used for biomonitoring of air quality. Species like SaralaThambavani D and Prathipa V. 2012. Bio
Azadiracta indica, Delonix regia, Moringa tinctoria, monitoring of air pollution around urban and industrial
Calotropis gigantea, Thyme rosemary and Cynodon sites. Journal of research in Biology 1:007-014.
dactylon can be potentially used for biomonitoring of air
Sarala Thambavani D, Sabitha MA. 2012. Water
quality in polluted areas.
quality and environmental assessment of sugar mill
effluent. Journal of research in Biology 2: 125-135.
REFERENCES
Chattapadhyay SP. 1996. Leaf surface effects of air Sarala Thambavani D, Saravana Kumar R. 2011.
pollution on certain tree species in Calcutta. Advance The monthly changes of chloroplast pigments content in
Plant Sci., 9(1):1-14. selected plant species exposed to cement dust pollution.
Journal of research in Biology 8:660-666.
Choudhury P and Banerjee D. 2009. Biomonitoring of
Air Quality in the industrial Town of Asansol using the Singh SK. 1991. Air Pollution Tolerance Index of plants.
Air pollution Tolerance index Approach. Res. J.of Chem Journal of Environmental Management. 32:45-55.
and Env., 13(1):46-51.

Tiwari S, Bansal S and Rai S. 1993. Air Pollution
CPCB. 1999-2000. Guidelines for developing green Tolerance Index of some plants in Urban areas of
belts. CPCB Publication. Programme objectives series Bhopal. Adv. Ecological., 16(1):1-8.
PROBES/75,
Treshow M. 1985. In: Air pollution and plant life,
Gonzalez L. Gonzalez-Vilar M and Reigosa MJ. 2001. JohnWiley and Sons, Newyork.
Determination of relative water content. Handbook of

plant Ecophysiology Techniques Kluwer Academic
Publishers. Dordrecht, The Netherlands. 207-212.
Hemmat Khattab. 2007. The defense mechanism of Submit your articles online at Ficuspublishers.com
Cabbage plants against Phloem sucking aphid Advantages
(Brenicoryne brassica L) Australian Journal of Basic and
Applied Science. 1(1):56-62. Affordable Charges
Quick processing
Mashita PM and Pise VL. 2001. Biomonitoring of air Extensive indexing
pollution by correlating the pollution tolerance index of
some commonly grown trees of an urban area Poll.
Research 20(2):195-197. www.ficuspublishers.com/submit1.aspx.

Effect of feeding frequency on growth performances and survival of

Rutilus rutilus caspicus
Authors: ABSTRACT:
Majid Mohammad Nejad
Shamoushaki. Feed management plays a major role in the economical and environmental
status of fish farms. Optimum feeding frequency seems to be dependent on fish size
and higher frequency of feeding was found to be advantageous for higher growth and
survival in younger age groups. The fishes should have the access to feed up to
Institution:
satiation for their optimum growth. This experiment was conducted for six weeks at
Department of Fishery,
four treatments and three replications as follows: Treatment A: One time a day at 8:00
Bandar Gaz Branch, Islamic
Azad University, Bandar h, treatment B: Two times a day at 08:00, 12:00 h, treatment C: Three times a day at
Gaz, Iran. 08:00, 12:00, 16:00 h and treatment D: Four times a day for feeding at 08:00, 12:00,
16:00, 20:00 h. Given the importance of the physical and chemical factors and their
impact on water supply and ultimately the fish growth, these factors were so
controlled through the experiment that the amount of dissolved oxygen was fixed on
5.5 - 6 ppm, the temperature 26 2 0C and pH 7.5 to 8. Rutilus rutilus caspicus were
Corresponding author: fed during the experimental period with SFK feed staff containing: 8.7 % moisture,
Majid Mohammad Nejad 11.2 % ash, 32 % protein and 10.5 % fat. Feeding rate was 10% of total body weight of
Shamoushaki. fingerlings. According to the results of this study, it was shown that feeding frequency
no affect body weight and length gain of fish and there is no significant difference in
the rate of FCR, BWI%, SGR, GR and CF (p < 0.05). Result of this study showed that
increasing of feeding frequency there isnt effect on growth and survival in Rutilus
rutilus caspicus.
Email: Keywords:
Maijd_m_sh@bandargaziau.ac.ir. Feeding frequency, growth performances, survival, Rutilus rutilus caspicus.
Article Citation:
Web Address:
Majid Mohammad Nejad Shamoushaki.
Documents/RA0212.pdf. Effect of feeding frequency on growth performances and survival of
Rutilus rutilus caspicus.
Dates:
Received: 10 Mar 2012 /Accepted: 22 Mar 2012 /Published: 04 Apr 2012
Ficus Publishers.
cited.
200-205 | JRB | 2012 | Vol 2 | No 3

Majid Mohammad Nejad Shamoushaki, 2012
INTRODUCTION excess feeding (Ng et al., 2000; Mihelakakis et al., 2002;

Rutilus rutilus caspicus belongs to Cyprinidae is Webster et al., 2002). In this context, it is useful to know
one of the most economically important and valuable the optimum feeding rate of the cultured species and how
telostei in the Caspian Sea. This kind of fish exist in the feed efficiency, feed consumption and composition of
southern part of the Caspian Sea especially Iran's shores. flesh are affected by it (Eroldogan et al., 2004). Among
Approximately two third of the semi artificial the different feed management practices proven to
propagation in Golestan province was done. Fish is the maximise the benefit of feeding, feeding frequency and
major source of protein for over one billion people ration size play an important role in regulating the feed
around the world (Rameshguru et al., 2011). intake, growth and waste outputs of fish (Silva et al.,
Understanding of natural foods and dietary habits of fish 2007). Optimizing feeding frequency may minimise feed
culture could be an important factor in providing wastage, leading to improvement in culture environment
effective method of nutrition. Although intensive fish and or reduction in size heterogeneity (Dwyer et al.,
culture adaptability of the species with different feeding 2002; Tucker et al., 2006), whereas poorly timed or
methods have been proven, but the choice of methods to sporadic feeding frequency may lead to increased
provide food and nutrition in aquaculture should be hunger, intra-specific aggression and increased rate of
considered dietary at patterns of normal behavior (Afshar cannibalism (Folkvord and Ottera, 1993). Studies on
Mazandaran, 2002). In result, understanding the feeding behavior in several fish species have shown that
nutritional qualities of fishes are very important. Fish if feeding frequency be in accordance with natural
feeding is one of the most important factors in feeding, it will increase growth and decrease FCR
commercial fish farming because feeding regime may (Bolliet et al, 2001). So, with aspect to two or three times
have consequences on both growth efficiency and feed feeding per day in farms and existing food for Rutilus
wastage (Tsevis et al., 1992; Azzaydi et al., 2000). The rutilus caspicus feeding in Iran it seems essential to
growth of fish at all stages is largely governed by the specify the best feeding frequency for reaching the
kind of food, ration, feeding frequency, food intake and highest rate of growth on Rutilus rutilus caspicus.
its ability to absorb the nutrients. Among these, feeding Considering the ideas mentioned above in this research,
frequency is an important aspect for the survival and the effects of feeding frequency on the growth
growth of fish at the early stage (Mollah and Tan, 1982). performances and survival of Rutilus rutilus caspicus
Optimum feeding frequency seems to be dependent on were studied.
fish size and higher frequency of feeding was found to be
advantageous for higher growth and survival in younger MATERIALS AND METHODS
age groups. The fishes should have the access to feed up In this study has been carried out in Sijual bony
to satiation for their optimum growth. However, over- fishes reproduce and cultivate center (Gorgan, Golestan,
feeding leads not only to reduction in feed conversion Iran) on 2010 summer. This experiment was conducted
efficiency and increase in input cost, but also for six weeks at four treatments and three replications as
accumulation of wastes that adversely affects the water follows: Treatment A: One time a day at 08:00 h,
quality (Biswas et al., 2006). Moreover, knowledge of treatment B: Two times a day at 08:00, 12:00 h,
the optimum feeding rate is important not only for treatment C: Three times a day at 08:00, 12:00, 16:00 h
promoting best growth and feed efficiency, but also for and treatment D: Four times a day for feeding at 08:00,
preventing water quality deterioration as a result of 12:00, 16:00, 20:00 h. Initial body weight and length
average were 0.9 gr. Given the importance of the 2003. All data were analyzed with one-way analyses of
physical and chemical factors and their impact on water variance (ANOVA) and significant means were
supply and ultimately the fish growth, these factors were subjected to a multiple comparison test (Duncan) at
so controlled through the experiment that the amount of P<0.05. When the normality of data did not present, the
dissolved oxygen was fixed on 5.5 - 6 ppm, the nonparametric test Kruskal-Wallis to compare treatments
temperature 26 2 0C and pH 7.5 to 8. Rutilus rutilus and test Mann - Whitney for paired comparison between
caspicus were fed during the experimental period with treatments were used.
SFK feed staff containing: 8.7 % moisture, 11.2 % ash,
32 % protein and 10.5 % fat. Feeding rate which paid RESULTS AND DISCUSSION
attention to live weight and in different times and after Final weight and length of Rutilus rutilus
each two weeks biometry, equall 10 % of body weight is caspicus in different treatments with 1, 2, 3 and 4
calculated and was intered to each aquarium. Fish feeding frequency per day are shown in table 1. Obtained
performances were evaluated in terms of Feed results in this study showed that increasing feeding
Conversion Ratio (FCR), Specific Growth Rate (SGR, % frequency has no effect on increasing weight and length
d1), Body Weight Index (BWI %), Growth Rate (GR, of Rutilus rutilus caspicus and there is no need full
gr d1), Condition Factor (CF, gr/Cm) and Survival (%). difference in this respect among considered treatments (p
These performance indices were calculated as follows > 0.05). Comparison average of different feeding
(Hung et al., 1989; Ronyai et al., 1990; Biswas et al., frequency effects on Rutilus rutilus caspicus growth
2010): factors during culture period are shown in table 2. The
FCR=total feed intake/ total biomass gain results showed that there isn't any meaningful difference
SGR=[(ln final weightln initial weight)/ rearing in FCR, % BWI, SGR, GR, CF and survival in different
duration in days]100 treatments (p > 0.05).
BWI=[(body weight finalbody weight initial)/ body Culture condition like flock density, temperature,
weight initial]100 water quality and feeding frequency are effective on fish
GR=(body weight finalbody weight initial)/ rearing growth in aquaculture (Wallace et al, 1988). Chang in
duration in days some factors like feeding frequency, feeding technique,
BWI=[(body weight /total length3)]100 or fish density may cause some changes in different fish
Survival=(number of fish harvested/number of fish species body weight (McCarthy et al, 1996). Feeding
stocked)100 frequency is an important aspect for the survival and
For analysis of all data were used SPSS version growth of fish at the early stage (Mollah and Tan, 1982).
13 and a software program for drawing graphs of Excel Over-feeding leads not only to reduction in feed
Table 1.The average of weight and length of Rutilus rutilus caspicus in different treatments
Treatments Initial weight (gr) Initial length (cm) Final weight (gr) Final length (cm)
A 1 time feeding per day 0.90.23a 4.610.18a 3.250.087a 5.130.37a
B 2 times feeding per day 0.90.23a 4.610.18a 3.290.034a 5.060.33a
C 3 times feeding per day 0.90.23a 4.610.18a 3.270.077a 5.150.3a
D 4 times feeding per day 0.90.23a 4.610.18a 3.270.071a 5.120.28a
The small Latin letters show that there are significant differences among different treatments

Table 2. Effect of different feeding frequency on growth performances in Rutilus rutilus caspicus
Indicatores Treatment A Treatment B Treatment C Treatment D

FCR 3.730.1a 3.700.04a 3.710.04a 3.780.08a
SGR (% d1) 2.620.044a 2.650.017a 2.630.021a 2.630.035a
BWI (%) 2255.92a 2292.6a 227.42.78a 227.134.9a
GR (gr d1) 0.050.0017a 0.05130.0006a 0.05030.0006a 0.05030.0015a
CF (gr/Cm) 2.410.25a 2.540.12a 2.40.165a 2.460.34a
Survival (%) 93.332.89a 855a 93.332.89a 91.675.77a
The small Latin letters show that there are significant differences among different treatments
conversion efficiency and increase in input cost, but also 2002). Study results on young cat fish by Murai and
accumulation of wastes that adversely affects the water Andrews (1976) have shown that more feeding
quality (Biswas et al., 2006). Also, the survey results frequency is needed for growth increase. Similarly,
showed that increasing of feeding frequency there isnt Mollah and Tan (1982) and Charles et al (1984) have
effect on growth and survival in Rutilus rutilus caspicus. reported that increasing feeding frequency in Clarias
Booth et al (2008) noted that 1 to 4 feeding frequency macrocephalus and Cyprinus carpio will cause an
per day may have the best function for increasing growth increase in growth, the results of this study is not the
in Salmonidae and Australian snapper, the results of this same. Comparsion of other study show none of the
study is not the same. Johansen and Jobling (1998) have results were not consistent with the results of this study
reported that feeding frequency increase, fish swimming was that increased feeding frequency varies in different
activity increase too and so energy consuming will be fish.
more and growth will be less. The highest growth in the
low frequency of feeding occurs (Tsevis et al, 1992). Acknowledgements
Study conclusions have shown that one-time feeding will This research was supported by the Sijual bony
be enough for the normal growth of Micropoginiuas fishes reproduce and cultivate center (Gorgan, Golestan,
furnieri (Aristizabal-Abud., 1990), Korean rock fish (Lee Iran). I also thank Mr.Jabbare for supplying the Rutilus
et al., 2000), yellow tail flounder (Dwyer et al., 2002), rutilus caspicus fingerlings used in this study. I also
the results of this study is the same. But for some species thank Mr.Maleki, Mr.Shakiba and Mr.Eri for their
it should be 2 to 6 times a day like European seabass valuable help during the Experiments.
(Ruohonen et al., 1998) and Tilapia (Riche et al., 2004) ,
the results of this study is not the same.Read phonetically Conclusion
Research on business culture species like Black Rock However, research results showed that the feeding
fish have shown that one - time feeding per day results in frequency and growth rate are different in different
normal growth and full use of feed in comparison to one species.Listen Read phonetically Totally, we can
time feeding in every 2 days or 2 time feed per day conclude with respect to resulting conclusions that
(Guen- up et al, 2004) , the results of this study is not the feeding frequency per day will have a meaningful effect
same. Although results of other researchers showed that on Rutilus rutilus caspicus weight and length and two-
increasing feeding frequency, causes feed acceptance times feeding is advised under tested situation including
increase and fish growth in many cases (Dwyer et al., changing water and airing.
REFERENCES Dwyer K, Brown JA, Parrish C and Lall SP. 2002.

Afshar Mazandaran N. 2002. Scintific papers of Feeding frequency affects food consumption, feeding
aquatic food and drug supply and inputs. Publications pattern and growth of juvenile yellowtail flounder
Samarang. 216. (In persian) (Limanda ferruginea). Aquaculture, 213:279-292.
Aristizabal-Abud EO. 1990. Effect of feeding Eroldogan OT, Kumlu M and Aktas M. 2004.
frequency in juvenile croaker, Micropogonias furnieri Optimum feeding rates for European sea bass
(Desmarest) (Pisces: Sciaenidae). J. Fish Biol, 37:987- (Dicentrarchus labrax L.) reared in seawater and
988. freshwater. Aquaculture, 231:501-515.
Azzaydi M, Martines FJ, Zamora S, Sanchez- Folkvord A and Ottera H. 1993. Effects of initial size
Vazquez FJ and Madrid JA. 2000. The influence of distribution, day length and feeding frequency on
nocturnal vs. diurnal feeding condition under winter growth, survival, and cannibalism in juvenile Atlantic
condition on growth and feed conversion of European cod (Gadus morhua L.). Aquaculture, 114:243-260.
sea bass (Dicentrarchus labrax L.). Aquaculture,
Guen-Up K, Jo-Young S and Sang-Min L. 2004.
182:329-338.
Effect of feeding frequency and dietary composition on
Biswas G, Jena JK, Singh SK, Patmajhi P and growth and body composition of juvenile rockfish
Muduli HK. 2006. Effect of feeding frequency on (Sebastes schlegeli). Faculty of marine Biosience and
growth, survival and feed utilization in mrigal, Cirrhinus Technology Kangnung National University Gangneung:
mrigala, and rohu, Labeo rohita, during nursery rearing. 210-702.
Aquaculture, 254:211-218.
Hung SSO, Lutes PB and Storebakken T. 1989.
Biswas G, Thirunavukkarasu AR, Sundaray JK and Growth and feed efficiency of white sturgeon (Acipenser
Kailasam M. 2010. Optimization of feeding frequency transmontanus) sub yearling at different feeding rates.
of Asian seabass (Lates calcarifer) fry reared in net Aquaculture, 80:147-153.
cages under brackishwater environment. Aquaculture,
Johansen SJS and Jobling M. 1998. The influence of
305:26-31.
feeding regime on growth and slaughter traits of cage-
Bolliet V, Azzaydi M and Boujard T. 2001. Effects of reared Atlantic salmon. Aquac. Int, 6:1-17.
feeding time on feed intake and growth. In: Houlihan D;
Lee SM, Hwang UG and Cho SH. 2000. Effects of
Boujard, T: jobling, M. (Eds), food intake in fish. Black
feeding frequency and dietary moisture content on
well science cost Action 827, Oxford: 232-249.
growth, body composition and gastric evacuation of
Booth MA, Tucker BJ, Allan GL and Fielder D. 2008. juvenile Korean rock fish (Sebastes schlegeli). Aqu
Effect of feeding regime and fish size on weight gain, aculture, 187:399-409.
feed intake and gastric evacuation in juvenile Australian
McCarthy ID, Carter CG and Houlihan DF. 1992.
snapper, Pagrus auratus. Aquaculture, 282:104-110.
The effect of feeding hierarchy on individual variability
Charles PM, Sebastian SM, Raj MCV and Marian in daily feeding in rainbow trout, Oncorhynchus mykiss
MP. 1984. Effect of feeding frequency on growth and food (Walbaum). J. Fish Biol, 41:257-263.
conversion of Cyprinus carpio fry. Aquaculture, 40:93-300.

Mihelakakis A, Tsolkas C and Yoshimatsu T. 2002. Tevis N, Klaoudatos S and Conides A. 1992. Food
Optimization of feeding rate for hatchery-produced conversion budget in sea bass, Dicentrarchus labrax,
juvenile gilthead sea bream Sparus aurata. Journal of the fingerlings under two different feeding frequency
World Aquaculture Society, 33:169-175. pattern. Aquaculture, 101:293-304.
Mollah MFA and Tan ESP. 1982. Effects of feeding Tucker BJ, Booth MA, Allan GL, Booth D and
frequency on the growth and survival of catfish (Clarias Fielder D. 2006. Effects of photoperiod and feeding
macrocephalus Gunther) larvae. Indian J. Fish, 29 frequency on performance of newly weaned Australian
(1&2):1-7. snapper Pagrus auratus. Aquaculture, 258:514-520.
Murai T and Andrews J W. 1976. Effect of frequency Wallace JC, Kolbeinshaven AG and Reinsnes TG.
of feeding on growth and food conversion of channel 1988. The effects of stocking density on early growth in
catfish fry. Bull. Jpn. Soc. Sci. Fish, 42: 159161. Arctic charr, Salvelinus (L.). Aquaculture, 73:101-110.
Ng WK, Lu KS, Hashim R and Ali A. 2000. Effects of Webster CD, Thompson KR and Muzinic L. 2002.
feeding rate on growth, feed utilization and body Feeding fish and how feeding frequency affects sunshine
composition of tropical bagrid catfish. Aquaculture bass. World Aquac, 33:20-24.
International, 8:19-29.
Rameshguru G, Senthilkumar P and Govindarajan

B. 2011. Vermiwash mixed diet effect on growth of
Oreochromis mossambicus (Tilapia). Journal of research
in Biology, 5:335-340.
Riche M, Haley DI, Oetker M, Garbrecht S and

Garling DL. 2004. Effect of feeding frequency on
gastric evacuation and the return of appetite in tilapia
Oreochromis niloticus (L.). Aquaculture, 234:657-73.
Ronyai A, Peteri A and Radics F. 1990. Cross breeding

of starlet and Lena river sturgeon. Aquaculture, 6:13-18.
Ruohonen K, Vielma J and Grove DJ. 1998. Effects of

feeding frequency on growth and food utilization of Submit your articles online at Ficuspublishers.com
rainbow trout (Oncorhynchus mykiss) fed low-fat herring
Advantages
or dry pellets. Aquaculture, 165:111-121. Easy online submission
Silva CR, Gomes LC and Brandao FR. 2007. Effect of Affordable Charges
feeding rate and frequency on tambaqui (Colossoma Quick processing
Extensive indexing
macropomum) growth, production and feeding costs Open Access and Quick spreading
during the first growth phase in cages. Aquaculture, You retains your copyright
264:135-139. submit@ficuspublishers.com

Effect of drought stress on protein and proline metabolism in seven

traditional rice (Oryza sativa Linn.) genotypes of Assam, India.
Authors: ABSTRACT:
Chutia J, Borah SP,
Tanti B. Abiotic stresses can directly or indirectly affect the physiological status of an
organism by altering its metabolism, growth, and development. Many plant species
naturally accumulate proline and protein as major organic osmolytes when subjected
Institution: to different abiotic stresses. These compounds are thought to play adaptive roles in
1. Department of Botany,
mediating osmotic adjustment and protecting sub cellular structures in stressed
Darrang College,
plants. Different approaches have been contemplated to increase the concentrations
Tezpur -784001 Assam.
of proline like compounds in plants grown under stress conditions to increase their
2. Department of Botany, stress tolerance. Seven different traditional rice varieties of Assam were evaluated for
Gauhati University, their response to osmolyte production under physiological drought condition through
Guwahati - 781014 Assam. simulation at three levels of osmotic stress of 0.15 bar, 0.25 bar and 0.56 bar of
physiological drought initiated by polyethylene glycol (PEG 6000). Along with the
evaluation for osmolyte response the different components of genotypic variation for
Corresponding author: six different drought-sustaining characters in the seven rice varieties were also
Chutia J. substantiated. The results indicated that plant height and seed number have
significant genotypic coefficient of variability (GCV) and heritability. Verities like
Laodubi, Leserihali, Beriabhanga and Borah were screened out as the best drought
Email: sustaining variety.
cjnan@rediffmail.com.
Keywords:
Phone No: Abiotic stresses, proline, protein, osmolyte, genotypic coefficient of variabil-
+91-94350-82261. ity, heritability, traditional rice cultivar.

http://jresearchbiology.com/ Chutia J, Borah SP, Tanti B.
Documents/RA0208.pdf. Effect of drought stress on protein and proline metabolism in seven traditional rice
(Oryza sativa Linn.) genotypes of Assam, India.
Journal of research in Biology (2012) 2(3): 206-214
Dates:
Received: 03 Mar 2012 /Accepted: 15 Mar 2012 /Published: 07 Apr 2012
Ficus Publishers.
cited.
206-214 | JRB | 2012 | Vol 2 | No 3

Chutia et al., 2012
INTRODUCTION instrumental in enabling the cells to elicit the right

Rice genotypes are known to vary widely in their response (Holmberg and Bulow 1998, Kasuga et al.
responses to abiotic stresses. About forty-two biotic and 1999, Serrano et al. 1999, Hasegawa et al. 2000, Zhu
abiotic stresses affect rice production (Sarkar et al. 2001a, Prabhavathi et al. 2002, Rontein et al. 2002).
2006). Recent studies indicate this in part due to the While there are several reports of expression of a number
complexity of interactions between stress factors and of genes only under water stress, a much more
various molecular, biochemical and physiological comprehensive approach is to profile the total protein
phenomena affecting plant growth and development contents and kinds of protein under normal and stressed
(Zhu 2002). Alterations in internal water relations are conditions.
generally evaluated by investigating the relationships It has also been seen that many plant species
between water potential or its solute and turgor naturally accumulate protein and proline as major
components and relative water content (Cutler et al. organic osmolytes when subjected to different abiotic
1980). stresses. These compounds are thought to play adaptive
Simulation of drought stress by polyethylene role in mediating osmotic adjustment and protecting sub
glycol (PEG) indeed induced drought stress on the plants cellular structures in stressed plants. However, not all
(Jiang et al.1995) significant deviation from the control plants accumulate protein or proline in sufficient amount
continues to increase with the increasing solute potential to help averting adverse effects of abiotic stresses. It has
(s) (Ranjbarfordoei et al. 2000). PEG-6000 has long also been observed that some stress proteins are
been utilized as a reliable marker under laboratory synthesized during the dehydration stress (Singh 2003,
conditions for testing the drought tolerant genotypes. Ashraf et al. 2007). Thus, different approaches have been
This is because polyethylene glycol acts as a non- contemplated to increase the concentrations of these
penetrating osmotic agent resulting into increasing solute compounds in plants grown under stress conditions to
potential (s) and blockage of absorption of water by the increase their stress tolerance. The present investigation
root system (Chezen et al. 1995 and Jiang et al. 1995). thus is aimed at elucidating the drought sustaining
Therefore, PEG solutions are often used to induce water character of some traditional rice cultivars of Assam to
stress in higher plants (Ashraf and OLeary 1996). drought stress based on some of the investigations
Drought screening using some seed technological conducted previously by others. Since breeders are still
parameters has been found to be quite useful in a number looking for traits that are suitable for screening rice
of crops (Singh and Afria 1988) under laboratory germplasm for characters affecting plant water relations
conditions. This technique can be further extended to test under drought conditions (Jha et al. 1997). The lack of a
drought tolerance in other genotypes, (Karan Singh et al. reliable method for identifying stress tolerant genotypes
2001). and the magnitude of factors involved in tolerance to
Response to water stress in plants at the water stress makes it difficult to choose traits conferring
molecular level undoubtedly constitutes an area of major an advantage under such stressed conditions.
interest for a complete understanding of the process. The
major strategy for gaining such understanding is through MATERIALS AND METHODS:
the approach of proteomics. Differential expression of The present study, initially 12 verities were
genes under water stress conditions can reveal a picture considered viz., Bengunguti, Beriabhanga, Borah,
as to what are the biochemical pathways that are Jahinga, Kesamani, Kolajoha, Laodubi, Leserihali,
Chutia et al., 2012
Pattesari, Rangadaria, Sakuakumal and Solpuna. After of fresh air to the growing seedlings at regular intervals.
initial studies related to germination index (GI) in PEG The seeds soaked in PEG-6000 solutions were kept
initiated drought and the whole plant behaviour under under observation for 7 days and the germination index
three water regimes then subsequently only seven was calculated out. The number of germinating seeds
traditional varieties viz. Laodubi, Borah, Jahinga, were counted and continued up to seven days at a regular
Beriabhanga, Pattesari, Leserihali and Kolajoha of interval of 24 hrs.
Assam, India, were screened for their response to Collected data were analyzed for determining the
osmolyte production under physiological drought (i) seed germination index, (ii) leaf protein content and
condition simulated by PEG 6000. (iii) leaf proline content.
Three levels of osmotic potential () of The germination index (GI) was calculated by
0.15bar, 0.25bar and 0.56bar induced by PEG-6000 were using the formula as suggested by the Association of
used for simulation of physiological drought. Seeds of Official Seed Analysis (AOSA 1983).
the experimental rice varieties were treated with different GI = No. of germinated Seeds + No. of germinated Seeds + - - + No. of germinated Seeds
Days of first count Days of Second count Days of final count
solutions of PEG- 6000. After the PEG-6000 treatment,
the germination index was determined and the seedlings For estimation of protein and proline content,
were subsequently grown under three different water young leaves from 20 days old seedlings grown under
regimes- (i) normal irrigated condition considered as non osmotic potentials of - 0.15 bar, 0.25 bar and 0.56 bar
-stress (control), (ii) unirrigated water stress upland were taken. The proline content was assayed by the
condition and (iii) unirrigated water stress potted method described by Bates et al. (1973) and Chinard et
condition. al. (1952). For the experiment, 0.5 gm of freshly
The experiment was conducted in a randomized collected leaves were homogenized in 10ml of 3%
block design (RBD) with three replications. Hundred aqueous sulphosalicylic acid. Control sample was
healthy seeds each of the 7 different cultivars was pre consisted of leaves from seedlings grown in deionized
soaked in distilled water for 12 hrs. Forty eight pairs of water alone. The homogenate was filtered through
clean and sterilized petri plates were used for the Whatman No. 2 filter paper. 2ml of the filtrate was taken
experiment. In each replication there were 16 petri in a test tube and 2ml of glacial acetic acid was added to
plates. The presoaked seeds were first air-dried to it. To the mixture freshly prepared 2ml of acid ninhydrin
eliminate the surface water. They were then placed over was added. The final solution was subjected to heat for 1
blotting paper in the petri-plates and were allowed to hr in a boiling water bath. After one hour of boiling the
germinate aseptically under three different osmotic reaction was terminated by placing the test tube in an ice
potentials i.e., 0.15 bar, 0.25 bar and 0.56 bar using bath. Now to the test tube 4 ml of toluene was added and
appropriate concentration of PEG-6000 (Ranjbarfordoei stirred for 20 30 seconds. Subsequently, the toluene
et al. 2000). Deionised water was used for the control layer was separated and the final mixture was again
and applied similarly. At regular intervals of 12 hrs, 5-6 warmed to room temperature and the red colour (slightly
drops of different solutions of PEG-6000 were red colour) was measured at 520nm.
administered to the seeds in the petri plates. The treated A standard curve was prepared using 0.1, 0.2,
and controlled seeds were allowed to germinate in a 0.3, 0.4 and 0.5 mol of pure proline and used for
BOD incubator at 25 + 20C for seven days. The lid of conversion of absorbance values into proline content.
the petri-plates were opened and replaced for exchange Protein content in the leaf samples was determined by
Journal of Research in Biology (2012) 2( 3): 206-214 208

Chutia et al., 2012
following Lowrys method (Lowry et al. 1951).
1240.633**
0.285
77155.4**
454.1803
Number
Proline and protein content were estimated from
Seed
the seedlings grown under simulated drought condition
Table 1 Mean sum of squares for various plant characters in twelve traditional rice cultivars grown under three different water regimes
induced by - 0.15 bar, 0.25 bar and 0.56 bar of PEG
6000. Seeds grown under osmotic stress induced by 0.56
bar of PEG 6000 failed to yield sufficient number of
Panicle
534.565**
length
15.16**
12.2341
seedlings enough for the biochemical assays. Thus
0.23
proline and protein could be estimated only in those
seedlings grown under 0.15 bar and 0.25 bar of artificial
Green leaf
Duration
22164.48**
drought.
571.383**
156.0795
Plant Characters
The genotypic and phenotypic coefficients of
0.037
variabilities for the characters were calculated according
to the formulae of Burton (1952). The heritability in
2238.785**
Flag leaf
broad sense was estimated according to Johnson et al.
angle
555.36**
221.2238
(1955 a, b).
0.675
RESULTS AND DISCUSSION
Flag leaf
Germination and seedling development under
length
163.01**
982.11**
57.25663
laboratory conditions have been accepted as suitable
1.0275
growth stages for testing the response to abiotic stresses
(Sharif-zadeh and Mohsen 2008). A positive correlation
between germination index (GI) in PEG initiated drought
1154.932**
24770.58**
height
Plant
438.9389
and the whole plant behaviour under three water regimes
0.194
were observed in the present investigation (Table 1).

This was evident from the results exhibited by Laodubi,
Degrees
freedom
Leserihali and Pattesari with higher germination index

of
while these same varieties showed good response to

11
92
2
other drought sustaining characters under three water

Germination
Character
regimes. Thus the determination of germination index

studied
196.40**
562.90**
(GI) can be used just as an easy and reliable parameter

index
0.00
6.59
for measuring drought sustenance among the traditional

rice cultivars of Assam.
Degrees of
The low germination rate in Jahinga, Pattesari

**Significance level P=0.1
Freedom
and Kolajoha as observed in the present study was due to

127
the osmotic stress induced by PEG 6000 which had mark

11
effect in both shoots and roots parameters. The reduction

Replication
in seed germination may be due to the less availability of

Variation
Source of
condition
Variety
Culture
free water to the seeds during early hours of inbibition,

Error
thus leaving the hydrolytic enzymes inactive (Shah and

Chutia et al., 2012
Loomis 1975, Hadas 1976). Inhibition of germination at rice plant. Ashraf and Foolad (2007) had reported that
higher osmotic potential may possibly be attributed to higher protein content in tolerant genotypes under water
moisture deficit in the seed below the threshold stress condition is due to higher DNA and RNA content,
requirement for germination (Everiff 1983). The which stimulate synthesis and inhibit protein
reduction in shoot and root growth is important as PEG decomposition.
induced stress affects root volume and root length Decrease in osmotic potential under stress
(Midaoui 2003). The reduction of root volume under reflects the increased hydrolysis of macromolecules into
induced osmotic stress originates not only from growth simpler ones like mono- and disaccharides, amino acids
inhibitions but also from a loss of turgidity (Huck et al. specially proteins etc. and consequently higher osmolite
1970). concentration (Tyagi et al. 1999). Thus under higher
Total protein content decreases due to abiotic solute potential, Laodubi, Leserihali, Beriabhanga and
stress Baruah et al. (1998). As synthesis of proteins occur Pattesari accumulated higher proline (Table 2), which
during dehydration stress a class of proteins called late acted as a osmoticum and accounted for higher drought
embryogenesis abundant globular protein known as tolerance due to greater relative water content and leaf
osmotin or dehydrin (Singh, 2003) are known to water potential (Baruah et al. 1998). This is because
accumulate in dry seeds, which play an important role in proline is a major organic osmolyte that accumulates in a
the regulation of dehydration in seeds. The protein variety of plant species in response to environmental
content among all tolerant genotypes was found higher stresses such as drought. Although their actual roles in
than susceptible ones (Serraj and Sinclair 2002). Water plant osmotolerance remain controversial, it is thought to
stress condition caused a marked change in protein have positive effects on enzyme and membrane integrity
synthesizing apparatus of plant tissue (Genkel et al. along with adaptive roles in mediating osmotic
1967) and the capacity for protein synthesis also adjustment in plants grown under stressed conditions.
decreases considerably as observed in response to water Exogenous application of proline to plants, before,
stress (Hsiao 1970). In the present study the results during, or after stress exposure, has been shown to
obtained with higher protein content in Borah, increase the internal levels of these compounds and
Beriabhanga, Laodubi and Solpuna (Table 3) are in generally enhances plant growth and final crop yield
agreement with the findings of Chinoy et al. (1974) who under stress conditions (Ashraf and Foolad 2007).
also reported a high protein content in drought stressed
Table 2 Proline content (mol/g of leaf tissue) in seven different rice cultivars
grown under simulated physiological drought stress
Culture condition
Simulated osmotic
Sl. No Variety Control Simulated osmotic
drought of - 0.15
(Deionized water) drought of - 0.25 bar
bar
1 Laodubi** 0.003 0.132 0.253
2 Borah** 0.001 0.0131 0.161
3 Jahinga 0.0003 0.173 0.145
4 Beriabhanga** 0.061 0.068 0.171
5 Pattesari 0.069 0.079 0.135
6 Leserihali** 0.057 0.053 0.204
7 Kolajoha 0.052 0.075 0.083
** Varieties selected as the best performing ones
Chutia et al., 2012
Table 3 Total leaf protein content (mg/g of leaf tissue) in seven different rice cultivars
grown Under simulated physiological drought stress condition
Culture condition
Sl. No Variety Control Simulated osmotic Simulated osmotic
(Deionized water) drought of 0.15 bar drought of 0.25 bar
1 Laodubi 0.12 0.16 0.15
2 Borah 0.15 0.18 0.14
3 Jahinga 0.12 0.19 0.15
4 Beriabhanga 0.11 0.21 0.15
5 Pattesari 0.119 0.18 0.13
6 Leserihali 0.15 0.19 0.12
7 Kolajoha 0.12 0.16 0.11
Over all water loss causes increase in The different components of genotypic variation
concentration of solutes leading to high concentration of for six drought-sustaining characters in seven
cell sap and intercellular fluid causes a greater decrease experimental rice varieties indicates that plant height and
in the water potential of the fluids. This causes stress on seed number have less environmental influences with
the protoplasm. Thus most of the biochemical processes high GCV as 187.16 and 35.99 respectively with high
are adversely affected because of water imbalances heritability (Table 4).
(Bunting et al. 1998). From this screening procedure Laodubi,
Tolerance to abiotic stresses is very complex at Leserihali, Beriabhanga and Borah cultivars were
the whole plant and cellular levels (Foolad 1999a,b, screened out as the best drought sustaining variety
Foolad et al. 2003a,b, Ashraf and Harris 2004). Putting among the ones considered in this investigation.
these observations under consideration the subsequent Mean values of the drought sustaining characters
phases of analysis was done so as to establish the in seven experimental rice verities are presented (Table
complexity of interactions between stress factors and 1). Genotypic components of variation for the traits are
various molecular, biochemical and physiological shown (Table 4).
phenomena affecting plant growth and development
(Zhu, 2002).
Table 4 Estimates of different genetical parameters in seven different rice varieties.

Genotypic Phenotypic
co-efficient co-efficient
Plant Genotypic Phenotypic
of variability
Heritability
Mean + SE Range variance Variance
of variability
characters GCV % PCV % %
Plant height 107.4286 129 - 55 40427.44 40521.63 187.16 187.38 99.76

Flag leaf
26.44444 50 - 11 55.09 77.91 28.06 10.31 70.71
length
Flag leaf
65.38095 90 -51 104.53 219.88 15.63 22.68 47.54
angle
Green leaf
111.3016 138 - 73 87.65 109.26 8.41 9.39 80.23
duration
Panicle
19.5873 24 - 11 4.49 11.41 10.83 17.25 39.39
length
Seed
70.80952 146 - 5 649.40 769.26 35.99 39.17 84.42
Number
Chutia et al., 2012
REFERENCES Chezen O, hartwig W, Newman PM. 1995. The

Ashraf M and Harrish PJC. 2004. Potential different effects of PEG-6000 and NaCl on leaf
biochemical indicators of salinity tolerance in plants. development are associated with differential inhibition of
Plant Sc., 166:3-16. root water transport. Plant Cell. 18:727-735.
Ashraf M. Foolad MR. 2007. Role of glycine betaine Chinard EP. 1952. J Biol Chem., 199:91.
and proline in improving plant abiotic stress resistance.
Chinoy JJ, Singh YD and Gurumurti K. 1974.
Env. And Experimental Botany 59:206-216.
biosynthesis of ascorbic acid and mobilization patterns of
Ashraf M. Leary O JW. 1996. Effect of drought stress macromolecules during water stressing germinating
on growth, water relations and gas exchange of two lines Ciser seedling . Biol. Plant. (Prague) 16:301-307.
of sunflower differing in degree of salt tolerance. Int. J.
Cutler JM, Sahan KW and Steponkus PL. 1980.
Plant. Sci., 157:729-732.
Alteration of the internal water relations of rice in
Association of Official Seed Analysis (AOSA). 1983. response to Drought hardening . Crop Sci., 20:307-310.
Seed Vigor Testing Handbook. Contribution No. 32 to
Everiff JH. 1983. Seed germination characteristics of
the handbook on Seed Testing.
three weedy plants species from south Taxas. J.Range.
Association of Official Seed Analysis (AOSA). 1991. Mange. 36:246-249.
Rules for testing seeds. J. Seed Technol., 12:18-19.
Foolad MR. 1999b. Comparison of salt tolerance during
Baruah KK and Singh K and Kumar A. 1998. seed germination and vegetative growth in tomato by
Evaluation of Drought tolerance in groundnut Stress and QTL mapping. Genome 42:727-734.
Environmental Plant Physiology Pointer Publishers,
Foolad MR, Zhang L, Subbiah P. 2003a. Genetics of
Jaipur, Rajasthan, India 145-152.
drought tolerance during seed germination in tomato:
Baruah KK, Bhuyan SS, Ghosh TJ and Pathak AK. inheritance and QTL mapping. Genome. 46:536-545.
1998. Response of rice genotypes to moisture stress
Foolad MR, Subbiah P, Kramer C, Hargrave G, Lin
imposed at seedling stage. Indian J. Plant Physiol., 3
GY. 2003b. Genetic relationships among cold, salt and
(3):181-184.
drought tolerance during seed germination in an
Bates LS, Walden RP and Teare ID. 1973. Plant Soil. interspecific cross of tomato. Euphytica. 130:199-206.
39:205.
Genkel PA, Satarova NA and Tvorus EK. 1967. Effect
Bunting AH and Kasam AH. 1988. Principles of crop of drought on protein synthesis and the state of
water use, dry matter production that govern choice of ribosomed in plants . Fiziol, Rast., 14:898-907.
crops and systems. In:Drought research priorities for the
Hadas A. 1976. Water uptake and germination of
dryland Tropics. (Eds.) bidinger, F. R. and Johansen, C
leguminous seeds under changing external water
ICRISAT, Pathancheru, 43-61.
potential in osmoticum solution . J.Exp.Bot., 27:480-489.
Burton GW. 1952. Quantitative inheritance in grasses.
Hasegawa PM, Bressan RA, Zhu JK, Bohnert HJ.
Proc. 6th Intern, Grasslands Congr., 1:277-283.
2000. Plant cellular and molecular responses to high
salinity. Annu. Rev. Plant Physiol. Plant Mol. Biol.,

Chutia et al., 2012
51:463-499. Lowry OH. Rosenbrough NJ. Farr AL. Randall RJ.

1951. Protein measurement with folin phenol reagent. J.
Holmberg N, Bulow L. 1998. Improving stress
Biol. Chem., 193:265-275.
tolerance in plant by gene transfer. Trends Plant Sci.,
3:61-66. Midaoui M, El. Serieys H and Kaan F. 2003. Effects
of osmotic and water stresses on root and shoot
Hsiao TC. 1970. Rapid changes in the levels of
morphology and seed yield in sunflower(Helianthus
polyribosomes in Zea mays in reponse to water stress.
annuus L.) genotypes breed for morocco or issued from
Plant Physiol., 46:281-285.
introgression with H. argophyllus T. & G. and H. debilis
Huck MG. Kleper B and Taylor MM. 1970. Diurnal Nutt. HELIA. 26. Nr. 38:1-16.
variation in root diameter. Plant Physiol., 45:529-530.
Prabhavathi V, Yadav JS, Kumar PA, Rajam MV.
Jha BN, Singh RA. 1997. Physiological responses of 2002. Abiotic stress tolerance in transgenic eggplant
rice varieties to different levels of moisture stress. Plant (Solanum melongena L.) by introduction of bactrial
Physiol., 2(1):81-84. mannitol phophodehydrogenase gene. Mol. Breeding
9:137-147.
Jiang Y, Macdonald SE, Zwiazak JJ. 1995. Effects of
cold storage and water stress on water relations and gas Ranjbarfordoei A, Samson R, Damne PV and
exchange of white spruce (Picea glauca) seedlings. Tree Lemeur R. 2000. Effects of drought stress induced by
Phsiol., 15:267-273. polyethylene glycol on pigment content and
photosynthetic gas exchange of Pistacia khinjuk and P
Johnson HW, Haward CL and Khan AR. 1955a.
mutica. Photosynthetic. 38(3):443-447.
Genotypic, Phenotypic correlations in soyabean and their
implication in selection. Agron. J., 47:477-483. Rontein D, Basset G, Hanson AD. 2002. Metabolic
engineering of osmoprotectant accumulation in plants.
Johnson HW, Robinson HF and Comstock RE.
Metab. Eng., 4:49-56.
1955b. Estimates of genetic and environmental
variability in soyabean. Agron. J., 47:314-318. Sarkar RK, Reddy JN, Sharma SG and Abdelbagi
MI. 2006. Physiological basis of submergence tolerance
Karan Singh and Kakralya BL. 2001. Seed
in rice and implication for crop improvement. Curr. Sci.,
physiological approach for evaluation of Drought
9(7):899-906.
tolerance in groundnut Stress and Environmental Plant
Physiology (Eds. K K Bora and Karan Singh and Arvind Serraj R. Sinclair TR. 2002. Osmolyte accumulation:
Kumar pp Pointer Publishers, Jaipur, Rajasthan, India. 45 can it really help increase crop yield under drought
-152. conditions? Plant Cell Env., 25:333-341.
Kasuga M, Liu W, Miura S, Yamaguchi-Shinozaki K, Serrano R, Culianz-Macia FA, Moreno V. 1999.

Shinozaki K. 1999. Improving plant drought, salt, and Genetic engineering of salt and drought tolerance with
freezing tolerance by gene transfer of a single stress- yeast regulatory genes. Sci. Hort., 78:261-269.
inducible transcription factor. Nat. Biotechnol., 17:287-
Shah CB and loomis RS. 1975. Ribonucleic acid and
291.
protein metabolism in sugar beet during drought. Plant
Physiol., 18:240-254.
Chutia et al., 2012
Sharif-zadeh and Mohsen J. 2008. Influence of priming

techniques on seed germination behavior of maize
inbreed lines (Zea mays L.) Journal of Agricultural and
Biological Science. 3(3):22-25.
Singh DP. 2003. Water deficit stress in Stress

physiology, New age Publishers, N Delhi. 64-79.
Singh K and Afria BS. 1988. Seed technological

approach for evaluation of drought tolerance in wheat
germplasm . Proc. Nation Semin.Seed Sci Tech. Ed. T.P.
Yadav and Changi Ram, HAU, Hissar., 72-178.
Tyagi A, Kumar N and Sairam RK. 1999. Efficacy o

RWC,membrane stability, osmotic potential, endogenous
ABA and root biomass as indices for selection against
water stress in rice. Indian J. Plant Physiol., 4(4):302-
306.
Zhu JK. 2001a. Plant salt tolerance. Trends Plant Sci.,

2:66-71.
Zhu JK. 2002. Salt and drought stress signal

transduction in plants. Annu. Rev. Plant Physiol. Plant
Mol. Biol., 53:247-273.

Advantages
Affordable Charges
Quick processing
Extensive indexing
Journal of Research in Biology(2012) 2(3) :206-214 214

Morphological alterations caused by Diflubenzuron in

Anopheles darlingi root (Diptera, Culicidae).
Authors: ABSTRACT:
Costa FM, Tadei WP.
The morphological alterations caused by Diflubenzuron, LC50= 0.006 ppm, in
larvae, pupae, and adults of Anopheles darlingi (Root 1926) were sistematized. The
Institution: third stage larvae showed elongation of the cervix 18-20 hours after exposure to the
Instituto Nacional de insecticide. Ecdysis in these larvae started after 40 hours: mortality began, with tissue
Pesquisas da Amaznia - extravasation and difficulties to discard the exuvia. The fourth stage larvae showed
INPA/CPCS. Laboratrio de tissue extravasation in the beginning of the formation of the puparium. The mouth
Malria e Dengue. Avenida parts of the larvae that completed the puparium were exposed, light-colored, and
Andr Arajo, 2936 presented clefts on the integument. Many of those that were able to perform ecdysis
Petrpolis CEP 69083-000 to adult could not escape the exuvia, and died bound by the legs, tarsi, and abdomen.
Manaus, Amazonas, Brasil.

Costa FM. Chemical control, IGR, larvicide, mosquito, larva.
Email: Article Citation:

fabiologocosta@gmail.com Costa FM, Tadei WP.
Morphological alterations caused by Diflubenzuron in Anopheles darlingi
Phone No: Root (Diptera, Culicidae).
55-92-3642-3435. Journal of research in Biology (2012) 3: 215-221
Web Address: Dates:

Ficus Publishers.
cited.
215-221 | JRB | 2012 | Vol 2 | No 3

Costa and Tadei, 2012
INTRODUCTION incapacity to discard the exuvia at ecdysis. Therefore,

The mosquito species Anopheles darlingi (Root immature stages are the most sensitive to this insecticide,
1926) is very important in public health because it is the because of their successive ecdyses that they undergo
main vector of the plasmodia that cause human malaria until reaching the adult stage (Eisler, 1992).
in Brazil. Although it is essentially a wild species, it Although the action mode of this compound is
proliferates rapidly in environments that are being altered already known in some insects, the response of each
by man to explore natural resources, such as for road species is an important information contributing to vector
opening, dam construction, aquaculture tanks, among control public policies concerning the larvicidal use of
others, where breeding grounds and human blood Diflubenzuron. Therefore, the objective of this study was
sources are available (Tadei et al., 2007). The control of to describe the morphological alterations caused by
this vector is the main measure to prevent the incidence Diflubenzuron in larvae, pupae, and adults of A. darlingi.
of human malaria. Currently, vector control has been
restricted basically to two forms: 1) Biological control, MATERIALS AND METHODS
by means of entomopathogenic bacteria of the species Obtaining the mosquitoes in the field and rearing
Bacillus sphaericus (Neide 1904), for larval forms, them in the laboratory
which is quite expensive; 2) Chemical control, with Females of A. darlingi were collected in two
pyrethroids, for adult forms (Tadei, 2001), applied localities in the periphery of Manaus, Amazonas State,
residually on walls or spacially by thermonebulization. Brazil: Puraquequara (338.63S; 595337.52W) and
In most places, chemical control has been the only Brasileirinho (3210.47S; 595217.22W). The
control measure undertaken. females were captured with an entomological aspirator,
It is known that the excessive and continuous use the bait being the authors with the body adequately
of only one control measure contributes strongly to protected by personal protective equipment - PPE. The
trigger resistance processes in populations of exposed collected females were kept in 350 ml capacity waxed
mosquitoes. For this reason, constant research is paper cups, covered with tulle. Next, they were taken to
necessary aiming at finding new chemical agents capable the laboratory, where they were fed domestic duck blood
of safely destroying populations of vector mosquitoes, and 10 % glucose solution. Upon feeding, they were
but it is crucial to understand before the mode of action individually transferred to plastic cups containing moist
of those agents and the responses of the different species filter paper, that served as substrate for oviposition. Once
exposed. the eggs ecloded, the larvae were raised according to the
Insect Growth Regulators (IGR) are known as method described by Scarpassa and Tadei (1990) until
third generation insecticides (Consoli and Loureno-de- they reached the optimal age for bioassys.
Oliveira, 1994). They are compounds of heteregeneous Bioassays
chemical nature with the capacity to interfere with Two experiments were conducted, one with
certain enzymes and hormones that regulate the larvae at the end of the third stage, another with larvae at
development of the immature stages of insects. the end of the fourth stage. Each experiment was
Diflubenzuron is an IGR and a powerful inhibitor repeated three times, each with 20 larvae in cups
of chitin synthesis that causes severe alterations in insect containing 50 ml of distilled water, food, and
ecdysis (Mulla et al., 1974). Basically, it acts on the LC50=0.006 ppm Diflubenzuron obtained previously
integument, leading to cuticle deformations and to the from the experiments of Costa and Tadei (2011). In the
experiment with fourth stage larvae, the cups were individuals that died in this process. There was a
covered with tulle to keep confined the adults that gradation in the attempt to perform the apolysis of the
possibly emerged. Both the experiments were exuvia. Some larvae died immediately in the beginning
accompanied by control groups that were subjected to of the process with the obstruction of the new integument
the same conditions, with the exception of the exposure and tissue extravasation, which prevented them to escape
to the insecticide. from the head exuvia. In other larvae, the head escape
Readings were taken every hour until 72 hours was observed; however, they died with the extravasation
after the beginning of the experiment, where the behavior of tissues of the thoracic region. In these latter, the
of the mosquitoes in the experiments were observed, thoracic regions integument was quite thin and
with the additional aim of collecting individuals markedly dilated. Finally, a third event occurred where
immediately after death or emergence. All dead the larvae were able to complete the apolysis, but the two
individuals were fixed in an adequate solution, and later last abdominal segments remained attached to the
observed with a stereoscopic microscope and exuvia. The larvae tried to perform swimming
photographed with a coupled digital camera. Some adults movements from the surface to the bottom of the cup to
were photographed in the experiment cup at the moment complete the apolysis of the exuvia, but did so slowly,
of emergence. External morphological alterations were and eventually died attached to the exuvia at the bottom
observed in the head, thorax, cephalothorax (pupae), of the cup. In the control group, the larvae developed and
apendages (adults), and abdomen of the mosquitoes, and fed normally until the end of the experiment.
recorded in a notebook.
RESULTS
Third stage larvae
In the first 18 to 20 hours after the beginning of
the experiment, we observed the elongation of the
cervical region (Figures 1, A and B), which connects the
cephalic region with the thorax. With this elongation, the
muscles had a greater difficulty to sustain the weight of
the cephalic region, which was considerably sclerotized.
The larva kept the thorax and abdomen in the natural
position (parallel to the water column), but had
difficulties to maintain its head in this position, keeping
it turned to the bottom of the cup. The mouth parts
movements were not interrupted, and the larvae kept
feeding normally. Alterations in the spiracular plate or in
color before or during death were not observed. Figure 1. Morphological alterations observed in
third stage larvae of Anopheles darlingi exposed to
Approximately 40 hours since exposure to the Diflubenzuron. A: beginning of ecdysis. Detail of the
insecticide, some larvae initiated the process of ecdysis. symptom of elongation of the cervical region; B:
beginning of apolysis of the cephalic region; C:
At this moment, the opening of the ecdysial line extravasation of tissue of the cephalic region. ce:
(Figures 1, B and C) in the head was normal in all the cervix; el: ecdysial line; te: tissue extravasation.

Fourth stage larvae however, the rest of the body, such as the thorax and
The subsequent larvae and stages showed the abdomen, remained inside the larval exuvia. At
patterns of morphological alterations in response to the this stage, a marked dilation of the cephalothorax
inhibitory action of Diflubenzuron. These alterations are (Figure 2C) and tissue extravasation in several
succinctly described here and grouped according to the degrees, from little to total cephalothorax
stage when the moulting events leading to mortality or destruction, was observed.
adult emergence occurred: C. Incompletely formed pupae. Light-colored
A. Larvae without apparent external morphological pupae, characterizing little melanization,
alterations. Larvae that probably died of natural incompletely formed, with no capacity to complete
causes, not showing any morphological feature that the apolysis of the exuvia, which remained attached
characterized any moment in the moult. Generally, to the last abdominal segments of the pupa (Figure
death with these characteristics occurred within 3A). At this stage, the pupae presented
around the first 24 hours until approximately 40 discontinuities in the formation of the integument in
hours. the region of the cephalothorax that covered the eyes
B. Larvae with morphological alterations in the pre- and the mouth and locomotory parts. Consequently,
pupal stage. Larvae that initiated the ecdysis process the appendages such as mouth parts, legs, wings,
with the initial formation of the pupa (Figure 2A), antennae, and practically all the ventral part of the
but only with the apolysis of the head. Mortality cephalothorax, were exposed in direct contact with
occurred within a period of 50 to 60 hours after the water (Figure 3B). Clefts in the integument (Figure
beginning of the experiment. The ecdysial line 3C) of these regions were observed, which probably
(Figure 2, B and D) opened normally, and in some favored the influx of water to the pupa, causing its
cases the cephalic region completed the apolysis; death. These alterations were recorded between 55
and 60 hours after the beginning of the experiment.
D. Completely formed pupae, attached to the larval
exuvia. Pupae without morphological alterations, but
the last segments still connected to the larval exuvia.
These pupae performed characteristic movements,
trying to escape from the exuvia, ascending or
staying at the surface to breathe; however, after
successive attempts, they finally died at the bottom
of the cup.
E. Completely formed pupae. Pupae that died with no
Figure 2. Morphological alterations caused by morphological alterations even in color.
Diflubenzuron in the beginning of pupation of
Anopheles darlingi. A: beginning of ecdysis, with F. Adults confined in the exuvia. Adults that did not
formation of the cephalothorax; B: apolysis of the complete the total apolysis of the exuvia, remaining
cephalic region with tissue extravasation; C: Detail
of the cephalothorax, with dilation and thickening of confined in it, in several degrees of emergence,
the thin integument; D: Dorsal view, showing detail varying from those with only the cephalic region
of the tissue extravasation and of the ecdysial line.
ctd: cephalothorax dilation; te: tissue extravasation; exposed to those completely emerged (Figure 4, A,
ctf: cephalothorax ormation; el: ecdysial line. B and C), but with hind tarsi attached to the exuvia
mosquitoes, and these are restricted to the genera Aedes

and Culex, possibly due to the ease of rearing in
laboratory and abundance in the environment, mainly
urban.
In this study, third stage larvae were quite
susceptible to Diflubenzuron in the applied
concentration. For larvae of C. quinquefasciatus in the
same stage and for the concentration of 0.0025 ppm,
Jakobs (1973) described a mortality around 95%; the
larvae presented a partial rejection of the exuvia at the
moment of the moult, subsequently causing death. For
larvae of A. aegypti of the same age exposed to 1.0 ppm,
Borges et al. (2004) reported destruction of the body
surface, with poorly defined abdominal segments.
Figure 3. Morphological alterations in pupae of According to the authors, at the concentration of 0.1
Anopheles darlingi derived from fourth stage
larvae exposed to Diflubenzuron. A: non- ppm, there were fewer alterations and better defined
melanized pupa attached to larval exuvia; B: body segments, but a slenderer and smaller body
Ventral view of a pupa highlighting appendage structure.
exposure; C: Side view of a pupa highlighting
the clefts in the integument and wing exposure. The regions most affected during the experiment
we: wing exposure; mpe: mouth parts exposure; were the head and the thorax of the larva, which are the
lge: leg exposure; lex: larval exuvia; tr:
body parts where the process of apolysis of the larva
integument cleft.
from the exuvia is initiated. These regions withstand an
(Figures 4B and 5B). In the surviving adults, i.e., intense pressure from the hemolymph and the body
those totallly emerged, the loss of the hind legs muscles to break through the exuvia and force the escape
(Figure 4D) or part ot them, because they remained of the larva during ecdysis. In this situation, the larva,
adhered to the pupal exuvia (Figures 4C and 5C), having had the formation of its new integument
was also observed. In the final emergence process of hampered by the insecticide, which probably reduced
these adults, the physical effort in the attempt of drastically the chitin content, is incapable to sustain the
completing apolysis during15 hours, on average, pressure exerted by the muscles and the hemolymph
until their death or emergence finally occurred, was (Eisler, 1992).
clearly observed. The morphological alterations described for the
G. Normal adults. Adults that completed emergence fourth stage larvae of A. darlingi are consistent with
and did not present morphological alterations, at those obtained by Arias and Mulla (1975) and Bridges et
least externally. The aptitude of these adults to fly al. (1977). In the former study, the authors summarize
inside the cup was excellent. seven categories of morphological alterations caused in
fourth stage larvae of Culex tarsalis treated with Altosid,
DISCUSSION of the benzoilfenilurea group, of which Diflubenzuron is
In the literature, there are studies with few also a member. Mortality was described for all stages,
reports of morphological alterations caused by IGRs in especially for the beginning of pupation and also for the

Figure 4. Morphological alterations observed in the Figure 5. Details of the structures attached to the
emergence of adults of Anopheles darlingi derived pupal exuvia observed as the adults of Anopheles
from fourth stage larvae exposed to Diflubenzuron. darlingi emerged. The adults derive from fourth stage
A: beginning of emergence; B: adult with legs larvae. A: abdomen of adult attached to the exuvia;
attached to pupal exuvia; C: adult emerging with leg B: tarsi locked inside the exuvia; C: legs broken and
broken and attached to pupal exuvia; D: completely attached to the exuvia (photo taken directly in the
emerged adult with missing hind legs (C and D: experiment cup). maab: male adult abdomen; pex:
photos taken directly in the experiment cup). aab: pupal exuvia; ble: broken leg; taf: tarsus attached to
adult abdomen; pex: pupal exuvia; ale: absent leg; the exuvia.
lef: leg attached to pupal exuvia; ble: broken leg.
pupa. The light-colored pupae were called albino, and deformations similar to those caused by methoprene,
those that presented exposed mouth parts were called another IGR. These results agree with the data obtained
elephantoid, due to the resemblance with an elephants in this study, in which exposure of the mouth parts and
head. locomotory appendages such as legs and wings in the
In this study, the characteristics were placed in adults was observed, as well as adults that died emerging
six groups, which were quite striking in most of the in several degrees of escape from the exuvia and adults
organisms evaluated. The light color of the pupae locked in the pupal exuvia.
resulted from the fact that mortality occurred in the very The morphological alterations produced by
beginning of the formation of the puparium, and Diflubenzuron in the larvae, pupae, and adults of A.
consequently the natural sclerotinization process that darlingi were quite variable and striking in all bioassays
causes the natural darkening of the integument did not conducted in this study. We attempted to sistematize
take place. The discontinuity of the integument exposed those deformations and the events that occurred to
the mouth parts being formed inside the puparium, which facilitate understanding. Probably, individuals from other
probably made the authors label them elephantoid. In mosquito populations or species will show several
the second study, the authors observed the morphological responses in variable periods of exposure to, and
alterations in the third and fourth stage larvae of A. concentrations of, the insecticide, which may cause
aegypti with the 5[[[55-(dimethyl-amino)-1- different degrees of morphological alterations. This
naphthalenyl]amino]-1,3-benzodioxole] IGR added to a generates the need of additional studies, such as internal
fluorescent compound denominated FIGR, and described anatomical observations at the tissue and cell levels, in
order to clarify the mode of action of Diflubenzuron in Scarpassa VM, Tadei WP. 1990. Biologia de
mosquitoes. anofelinos amaznicos. XIII. Estudos do ciclo biolgico
de Anopheles nuneztovari (Diptera, Culicidade). Acta
REFERENCES Amaz 20:95-118.
Arias JR, Mulla MS. 1975. Morphogenetic aberrations
Tadei WP. 2001. Controle da malria e dinmica dos
induced by a Juvenile Hormone Analogue in the
vetores na Amaznia. Anais da VII Reunio Especial de
mosquito Culex tarsalis (Diptera, Culicidae). J Med
Manaus da Sociedade Brasileira para o Progresso da
Entomol., 12:309-316.
Cincia. Manaus, Sociedade Brasileira para o Progresso
Borges RA, Cavasin GM, Silva IG, Arruda W, da Cincia - SBPC. 6.
Oliveira ESF, Silva HHG and Martins F. 2004.
Tadei WP, Rodrigues IB, Santos JMM, Rafael MS,
Mortalidade e alteraes morfolgicas provocadas pela
Passos RA, Costa FM, Pinto RC and Oliveira AEM.
ao inibidora do Diflubenzuron na ecdise de larvas de
2007. Entomologia e controle de vetores: o papel da
Aedes aegypti (Diptera, Culicidae). Rev Pat Tropic.,
entomologia no controle da malria. Rev Bras Med
33:91-104.
Trop., 40:23-26.
Bridges AC, Cocke J, Olson JK and Mayer RT. 1977.
Effects of a new Fluorescent Insect Growth Regulators
on the larval instars of Aedes aegypti. Mosq News.
37:227-233.
Consoli RAGB, Loureno-de-Oliveira R. 1994.

Principais mosquitos de importncia sanitria no Brasil.
Rio de Janeiro, Fiocruz 228.
Costa FM, Tadei WP. 2011. Laboratory toxicity

evaluation of Diflubenzuron, a chitin-synthesis inhibitor,
against Anopheles darlingi (Diptera, Culicidae). J Res
Biology 6:444-450.
Eisler R. 1992. Diflubenzuron hazards to fish, wildlife,

and invertebrates: a synoptic review. U.S. Fish and
Wildlife Service. Contaminant Hazard Review - Report Submit your articles online at Ficuspublishers.com
25:4-9.
Advantages
Jakob WL. 1973. Developmental inhibition of Complete Peer review
mosquitoes and the house fly by urea analogues. J Med Affordable Charges
Quick processing
Entomol., 10:452-455.
Extensive indexing
Mulla MS, Darwazeh HA and Norland RL. 1974.
Insect Growth Regulators: Evaluation Procedures and
Activity against Mosquitoes. J Econ Entomol., 67:329-332. www.ficuspublishers.com/submit1.aspx.

Effects of two pressed and extruded foods on the pigmentation of the

flesh and filleting yield of rainbow trout (Oncorhynchus mykiss)
Authors: ABSTRACT:
Mustapha ABA1,
Belghyti Driss 1,
Elkharrim khadija1, In order to compare the pigmentation of meat thread of rainbow trout
Benabid Mohammed2. Oncorhynchus mykiss, double pressed and extruded foods, an experimental test was
conducted in a fish farm. Comparison of two different formulation of food and energy
are performed in different energy requirements. Following this study two plans were
Institution:
formulated that extruded with 42% crude protein, 28% fat and 17% carbohydrate and
1. Biology and Health
Laboratory. Environmental 50 ppm of astaxanthin while pressed and 44.7% crude protein, 15% fat, 28.6
and Parasitology Team / carbohydrate and 40 ppm of canthaxanthin with 20.9 MJ of digestible energy 16.48 MJ
UFR Doctoral Parasitology and the initial average weight of trout with 474 g higher in two circular tanks of fresh
compared: Medical and water open circuit, each group was fed twice a day. After 60 days of experimentation
Veterinary Applications." the average weight for the final extruded feed was 759 g (60.12% weight gain) and
Sciences Faculty. Ibn Tofail pressed for the 724 g (52.74%). The best conversion rate was obtained with the feed
University. Knitra B.P. 133, extruded with1.17 against 1.56 with a survival rate of 98.85% and respectively
14000. Morocco. 97.72%.While for the pigmentation of the skin it was obtained at using the scale of
Salmofan 26,46 and 23,35 for the extruded and pressed respectively .Filleting yield
2. National Center of was 65.14% for the extruted diet and 60.64% for the pressed diet.
Hydrobiology and
Pisciculture (NCHP) Azrou
Morocco.

Mustapha ABA. Pelleted, extruted, nutrition,pigmentation,carotenoids, fillet yeild.

aba_mustapha@yahoo.fr. Mustapha ABA, Belghyti Driss, Elkharrim khadija, Benabid Mohammed.
Effects of two pressed and extruded foods on the pigmentation of the flesh and
filleting yield of rainbow trout (Oncorhynchus mykiss).
Phone No:
212661570488. Dates:
Web Address: Ficus Publishers.

http://jresearchbiology.com/ This Open Access article is governed by the Creative Commons Attribution License (http://
Documents/RA0201.pdf. creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-
cited.
222-231 | JRB | 2012 | Vol 2 | No 3

Aba et al., 2012
INTRODUCTION Carotenoids can not only help improve quality by

Diet plays a major role on the nutritional quality improving the color, but could also contribute to a better
and taste of the flesh of fish.One of the strengths of image in the minds of consumers of aquaculture
farmed fish is the quality control (Regost 2001), and products. In addition to their role pigmentosa, these
increasing the fat content of foods,that is followed by a compounds may have biological activities, some of
decrease in the ratio protein / digestible energy food which are fundamental precursor of vitamin A,
which, leads to improved growth performance and antioxidant or immuno-stimulating (Choubert and al.,
reduced nitrogen discharges. (Corraze 1999, Aba 2011). 2006), but most research conducted to date have focused
A special feature of aquaculture in relation to on their role as pigments.
fishing is that it is possible to obtain some control over Several factors may affect the pigmentation of
the quality of fish meat for consumption through the diet salmonids, including fish size, the genetic factors sex,
(Kolditz 2008). Numerous studies have shown that the age, time to Pigment supplementation, food composition,
composition of the food and its energy content directly source and concentration of carotenoids. Among them
affect the body composition of fish (Medale 2010, many there are other factors that can influence the
Kolditz 2008, Aba 2011). The quality of farmed fish pigmentation of the flesh of fish found food composition
includes both the ability to transform in particular the (Torrissen, 1985).
notions of performance gutting, filleting (Regost 2001) In addition to the pigmentation of the flesh, the
but also one of the qualities of the flesh of fish fillet; net yield is considered an important quality criterion for
pigmentation considered by the consumer as a factor in aquaculture technology, because fish fillet, as the main
visual quality (Choubert 1999), the color of the net is one edible portion of fish, hold the main economic and
of the most important factors motivating purchase and nutritional interest of fish production. Increasing fillet
acceptability of a product by the consumer (Buttle et al., yield, without any negative effect on flesh quality, is a
2001; Baker and Gunther, 2004, Liu et al., 2004). major challenge for fish farmers (Bugeon 2010)
The setting of carotenoid pigments in fish is increased net yield is related to muscle development
dependent on many factors among these factors that proportionately higher than other tissues. This muscle
there is a food. The fish are not able to synthesize development may impact on the quality of the flesh in
carotenoids de novo and must therefore be found in their terms of composition in addition to the technological
diet. (Choubert 1999). quality leading to the return on gutting and filleting as a
This pigmentation is due to food-borne low cut performance can lead to economic losses
carotenoids, mainly astaxanthin and canthaxanthin (Bugeon, 2006 ).
(Torrisenet and al., 1989), these compounds are added to It is in this context that this test is performed to
feed the fish and the synthesis of carotenoids compare the effectiveness of two food and extruded and
supplementation increases the cost of the food about pelleted food , and determine the interaction between
15% to 20% (Choubert 1999). Carotenoids are classified these foods their formulation and their effects on the
according to a color scale based on differences in pigmentation of trout fillets by supplementation of
astaxanthin, which gives a pink red and canthaxanthin, carotenoids, and yield of filleting.
which gives more of a yellow color (Knockaert, 2006).
Therefore, the color of the fish is a decisive quality
criterion that must be maintained and optimized.
Aba et al., 2012
MATERIALS AND METHODS site, we have set the rates according to the temperature of
experimental design the site which is about 14 so that the quantitative ratio
The experiment was conducted between April for the same food energy intake is: amount of food
15, 2009 and June 27, 2009 at the fish farm located about extruded 1.27 = amount of pressed food (or amount of
70 km from Azrou (Morocco). This test was conducted food extruded 0.78 = amount of pelleted food). Gross
in two circular tanks of 22 m3 volume at the open circuit energy was calculated using the following values: crude
with an initial load of 14 kg fed by spring water at a protein = 23.7 kJ g-1, crude lipids = 39.5 kJ g-1 and
constant temperature of around 14 C and a flow rate of carbohydrate = 17.2 kJ g-1 proposed by Brett and Groves
30 m3 h-1 , with a time of renewal of water 1.4 times per (1979). The calculation of digestible energy is obtained
hour with oxygen levels above 80% saturation. The by the coefficient of digestibility of protein, fat and
average content of dissolved oxygen in the outlet of the carbohydrates gelatinized or raw (Guillaume et al.,
ponds was 7.1 ppm. 2001).
biological materials Body Measurements
1400 juvenile trout females triploid of average Body mass, length, and organ mass were
weight 474 g from the same batch of eggs were divided recorded to evaluate the condition factor (CF) = [total
randomly into two circular tanks. The test was conducted body weight (g)] / [total body length (cm)] 3 ;
in monoculture, fish were fed manually and the daily
ration was split into two meals distributed at 09 AM and Table 1a : Ingredient and proximate compositions
(g/100 g dry matter) of the truted diets
03 PM, for 60 days, according to the feeding table
Ingrdients Extruted diet
provided by the supplier of food (71 Ecolife of Biomar). Fish meal 30%
Every two weeks 30 fish of each batch have been Soybean meal 15%
Rapeseed oil 12%
anesthetized after 24 h of fasting in order to measure the Fish oil 10%
size and the weight of each fish .for measurements of soy concentrate 8%
wheat 6%
weight and size.The quantities of food distributed were
Rapeseed meal 6%
weighed to estimate the consumption by the fish between Krill meal 5%
two weighings. Vitamin A - (UI/kg) 5000
Vitamin D3 (UI/Kg) 1000
experimental foods
Vitamin E - (mg/kg) 180
Formulation and chemical composition of
Vitamin C - (mg/kg) 100
experimental diets are shown in Table 1a,1b and 2. Astaxanthin 50 ppm
food composition ashes 7%
The rate of feeding Table 1b : Ingredient and proximate compositions
(g/100 g dry matter) of the pelleted diet.
The experimental test was aimed at comparing
two non-isoenergetic foods to different formulations on Ingrdients Pressed diet
their growth performance of fish, and their flesh quality Fish meal 41,5%
Fish oil 11,5%
in isoenergetic condition. The amount of food distributed
Corn gluten 15%
is consistent with the feeding tables of extruded and
Wheat Flour 30%
pressed foods that have different digestible energy 20.90 vitamin complex 2%
Mj ,16.48 Mj, respectively. These rates of rationing Cantaxanthin 40 ppm
depends on the temperature of the water colsely of the ashes 6,1%

Aba et al., 2012
viscerosomatic index (VSI) = ([viscera weight (g)] / determined by folch method (1957)
[total body weight (g)] x 100) (Ricker, 1979). Technical Variables
Zootechnical parameters Pigmentation of the flesh was done by the use of scale
Calculations: The following variables were rock SalmoFan
calculated: The perceived color of the nets was determined
Weight gain (WG, %) =100 x (final body weight - initial on fresh fillets in the processing plant using the color
body weight) / initial body weight scale, the SalmoFan (F. Hoffman-La Roche Ltd, Basel,
Survival(%) =100 x (final amount of fish /initial amount Switzerland). Three people have made separate
of fish) determinations for each fillet in natural light.
Average daily growth (g) = (final wt initial wt) /no. of Fillet yield was calculated as F%=( Fw/Bw) 100,
days where Fw was the fillet weight and Bw was the body
Feed conversion ratio (FCR) = g feed consumption/ (g weight in grams.
final biomass initial biomass). Statistical studies
Specific Growth Rate (SGR) = 100 (Ln final weight (g) - Our results are compared statistically (R
Ln initial weight (g)) / time (days). Development Core Team, 2011). All parameters of
Chemical analysis growth and yield were subjected to analysis of variance
Crude protein, crude fat and ash were determined test (ANOVA). The results were subjected to analysis of
from the ventral muscle, according to AOAC (1990). variance and any differences were estimated by the
Eight fillets of final fish carcasses and muscles Duncan test (1955) at the 0.05 level.
were sampled and stored at -25C for proximate
analyses,which were performed according to AOAC. RESULTS
Dry matter was determined after drying at 105C until a Our experimental test showed that the
constant weight was obtained. Ash content was measured performance of zootechnical parameters vary
by incineration in a muffle furnace at 525 C for 12 h. significantly (p <0.05) between the two dietary
Crude protein was analyzed by the Kjeldahl method after treatments (Table 3). Indeed the final weights of fish are
acid digestion using the Gerhardt system. Lipid was between 724 and 759 g for diets extruded and pressed .
Table 2 : Proximate composition of diets (%) Duncan's test showed a significant difference between
Extruted the final weights (p<0.05). The percentage weight gain
diet
Pressed diet
was 52.74 for the pelleted food, where as it was 60.12 for
Dry matter 96 % 93,2 %
Protins 42% 44.7 % extruded feed. There it showed a significant difference
Lipids 28% 15 % between the two values of the two systems (p<0.05). The
carbohydrates 17% 28.6 %
Fiber 2% 2% TCS was calculated as 0.7% for fish fed with the diet in a
Ash 6% 6,1 % pressed feed and 0.8% for the extruded diet, there was a
Moisture 4% 6.80 %
significant difference (p<0.05) between these results.
phosphorus 1% 1,15 %
Gross Energy The VI was 7.88 for the pressed food and 10.29 for the
24,28 21,58
(GE, Mj Kg-1)
extruted food.The survival rate was 97.72% for pelleted
Digestible energy
20,90 16,48
(DE, MJ Kg-1) food and 98.85% for the extruded food the difference
DP / DE (g MJ- 1)
18,08 24,41 between the two groups was not significant. The rainbow
(DP:Digestible Prtoein)
Ratio P /L 42/28 44,7/15 trout fed with extruded feeds had high levels of lipid in
Aba et al., 2012
Table 3 : Results of Rainbow trout performances performance (Aba and al, 2011) given that in salmonids,
obtained during this experimental test the optimal PD / ED in the order of 17-18mg/kJ
Pelleted Extruted
Paramters (Kaushik, 1997), which is lower when compared with
diet diet
Initial average weight (g) 474 474 that of later recommended 22-24mg/kJ value (Cowey,
Final average weight (g) 724a 759b 1992).
Weight gain (%) 52,74a 60,12b
Specific growth rate (%) 0,7a 0,8b The growth observed in this study represented by
Feed conversion ratio 1,56b 1,07a the weight gain are comparable to those obtained by
Viscerosomatic index 9,58a 10,99b
Survival (%) 97,72 98,85 Pfeffer and al. (1991) and those reported by Pokniak
(1999). These results are consistent with those reported
fillet compared with rainbow trout fed with pelleted by Zoccarato and al. (1996). With their experimental
feeds (p<0.05). The pigmentation of the fish fillet differ work concerning the trout, the pressed and extruded
(p<0.05) among fish fed with extruded and pelleted foods, the extruded food is high in fat, thus provides
feeds.The yield to evisceartion was 90,58 for pelleted more energy than the pressed food.
food and 89,61 for extruted food , the difference between Similar results were observed in the silver perch
two diets were significant.The filleting yield was 60,64 (silver perch Bidyanus bidyanus) by Booth (2002) and in
for the pelleted diet ,where as it was65,14 forthe extruted the Nile tilapia (Oreochromis niloticus) by Ammar
diet (p<0.05). (2008) whose body weight has been a big success win
with the extruded food compared to food in a hurry.
DISCUSSION Guroy (2006) work on the sea bass (Dicentrarchus
Increasing the level of digestible energy in fish labrax) with isoenergetic diets for pressed and extruded
feed incorporating a rate well digestible carbohydrates by was observed its weight and better performance of the
the extrusion treatment (Kaushik, 2000) and a high rate IVS to extruded food
of fat has resulted in improved animal performance. In addition, we can say that the diet can be
More over, this diet is a nutritional strategy applied to get extruded to have a greater incorporation of lipids, which
a savings of protein without compromising the growth of probably increased by the IVS increased visceral
trout (Cho & Bureau 2001). This sparing effect by fat,which is seen in the viscera that there are more
supplementation of fat and carbohydrates has been well deposition of fat in the rainbow trout (Corraze, 1999,
demonstrated for salmonids and sea bass (Watanabe, Richard, 2006; Kolditz, 2008; Medale, 2009) and IV
1982, Dias et al. 1998; Torstensen et al., 2001, Aba obtained in this study is almost consistent with the results
2011), perch Scortum barcoo Song and al., (2009) relates of Gelineau (2001).
the value of the digestible energy and digestible protein The increase in dietary energy intake (via the
levels, any decrease in the ratio PD / ED has a direct amount of food distributed or the energy content of food)
impact on the retention of proteins that is becoming more leads, in almost all species, with an increase in body fat
efficient, or the food pressed is characterized by a ratio accompanied by a decrease in water content (Dias, 1998,
of PD / ED high, this is found in the trout food poor Corraze and Kaushik, 1999, Medale 2010, Aba 2011),
Table 4 : Results of Rainbow trout technical variables obtained during this experimental test
Pigmentation
Paramtres Total lipid content of fillet Yield to evisceration Filleting yield
Scale of Salmofan
a
Aliment press 5,58 23,35a 90,58% 60,64 %a
Aliment extrud 8,35b 26,46b 89,61% 65,14 %b

Aba et al., 2012
this lipid accumulation in fish has been obtained in after processing of the product, and these results are
several species of fish such as rainbow trout (Caballero consistent with those of Torrissen (1986, 1989), and No
et al, 2002. Chaiyapechara et al, 2003), sea bass (the (1992) and (Baker et al., 2002). the effectiveness of
Pirin and Gatta, 2000) and Atlantic salmon (Hamre, astaxanthin is related to its high digestibility of up to
1998). The results obtained in this trial are consistent 60% while than that of canthaxanthin, it is only 30%.
with those obtained by Quillet et al (2007). According to Choubert (1999), in addition to the most
Salmonid pigmentation by carotenoids are effective dose for optimal retention of dietary pigments
affected by the dietary source of pigments, the rate of in Atlantic salmon 40-60 mg/kg of astaxanthin, rainbow
pigments, length of farming and food composition. trout in fresh water level ranged from 50 to 70 mg / kg of
Bjerkeng (2001) found that the food composition may be astaxanthinand this is consistent with our results.(Nickell
related to the manufacturing technology of the food that and Springate, 2001)
can lead to a food pressed or extruded above the For yielding we noticed that the technological
pigmentation of trout fillets which has a close performance of evisceration is almost 90% for the two
relationship with the lipid in food as food extrusion can foods but yield to evisceration of the pelleted food is
incorporate more fat (Aba et al., 2011) this increase in squeezed slightly higher compared to that of the extruded
dietary fat causes an increase in muscle lipid content and and this can not be explained by the large deposits of fat
increased color of the fillets. As a triploid trout , the in the viscera in trout fed the feed extruded (Medale,
pigmentation will have impact on the nets and skin color 2009) and our results are consistent with those of Quillet
(Choubert 1999). During this test we obtained the best (2007). While for the filleting yield there is an increased
pigment for the food extrusion since it contains high performance for the extruded diet, it is governed by a
levels of fat. Several studies have shown that it is better retention of protein due to the low ratio of
possible to increase the determination of carotenoids in digestible protein / digestible energy by the high rate of
the muscles of salmonids by altering the level of dietary fat found in food extruded and subsequently the protein /
lipids (Torrissen, 1989, Nickell and Bromage, 1998 (a) lipid. This decreases when the protein sparing effect and
Barbosa et al., 1999). And as the carotenoids are fat- subsequently a better retention of proteins resulting in
soluble compounds, their absorption and digestibility are better optimization of the use of proteins (Lee and Kim
related to the lipid content of the food (Choubert, 1999; 2001) and a better yield to the thread and our results are
Bronstad and al., 2002). Moreover, the binding of in agreement with those obtained by caballero et al
astaxanthin is better by compared to canthaxanthin. This (2002) and De Francesco and al (2004).
difference is attributed to better absorption of astaxanthin
in rainbow trout Rainbow (Torrissen,1989, Choubert CONCLUSION
1999). The mastery (control) of growth performance and
Similar results were observed in the same fish pigmentation of fish and technological performance are
(Nickell and Bromage, 1998; Choubert and Baccaunaud, related to the composition of the food and its method of
2006) and in Atlantic salmon (Bjerkeng et al. 1997; manufacture. The extruted feed offers the best trout
Bencze Rora, 1998; Einen and Thomassen, 1998) in growth performance by its energy content, buoyancy,
brown trout (Salmo trutta) (Regost et al., 2001). Rainbow digestibility and digestible protein ratio / digestible
trout uses astaxanthin more efficiently than energy and non-protein energy level factor. In intensive
canthaxanthin and astaxanthin is more stable during and fish farming using extruded diets despite their price can
Aba et al., 2012
be justified by the savings resulted from their feeding Baker R and Gunther C. 2004. The role of carotenoids
efficiency through better conversion and their in consumer choice and the likely benefits from their
contribution to sustainable aquaculture. In addition to inclusion into products for human consumption. Trends
feeding efficiency extruded food also contribute to in FoodScience and Technology, 15:484-488.
improve the quality of fish through improved
Bencze Rora AM. 2001. Primary processing
pigmentation which is closely related to the lipid content
(Evisceration and filleting). In "Farmed Fish Quality" (S.
in the food and also by a better performance due to
C. Kestin, and P. D. Warris, eds.). Blackwell Science,
thread better retention and better protein fat content that
Bristol, UK. 249-260.
allows a more attractive pigmentation of the net of the
rainbow trout. Bjerkeng B, Refstie S, Fjalestad KT, Storebakken T,
Rodbotten M, Roem AJ. 1997. Quality parameters of
ACKNOWLEDGEMENTS the flesh of Atlantic salmon (Salmo salar) as affected by
This research was financially supported by Les dietary fat content and full-fat soybean meal as a partial
Domaines agricoles Truite de l'Atlas Azrou .We thanks substitute for fish meal in the diet. Aquaculture 157:297-
manager and staff of fish farm 309.
Bjerkeng B. 2001. Carotenoid pigmentation of

REFERENCES
salmonids fishes recent progress.Avances en Nutrition
Aba M, Belghyti D, Elkharrim K, Benabid M,
Acuicola V Memorias del V Simposium Internacional de
Elguamri Y and Maychal A. 2011. Effects of two
Nutrition Acuicola. 19-22 Noviembre, 2000. Merida,
pressed and extruded foods on growth performance and
Yucatan.
flesh quality of rainbow trout (Oncorhynchus mykiss).
Sciencelib Editions Mersenne, volume 3 N111004. Booth MA, Allan GL, Evans AJ, Gleeson VP. 2002.
Effects of steam pelleting or extrusion on digestibility
Aba M, Belghyti D, Benabid M. 2011. Effects of two
and performance of silver perch Bidyanus bidyanus.
pressed and extruded foods on growth performance of
Aquaculture Research 33:1163-1173.
rainbow trout and their environmental impacts .
Sciencelib Editions Mersenne, volume 3 N111205. Brett JR, Groves TD. D. 1979. In fish physiology - VIII
(op. cit.) G physiological energetics, 280-282.
Ammar A. 2008. Effect of extruded and non extruded
fish pellet on growth performance and total production of Bugeon J, Vandeputte M, Cardinal M, Lefvre F,
nile tilapia and grey mullet fingerings reared in a Uyanik A, Davenel D, Labb L, Haffray P. 2006.
me
polyculture system in earthen ponds, 8 Symposium sur Amlioration des rendements au filetage et au parage en
laquaculture du tilapia October 12-14, 2008 Cairo, pisciculture TRUITAFILET . Rapport final de la
Egypt. convention OFIMER N076/04/C.
Baker RTM, Pfeiffer AM, Schner J, Lemmon-Smith Bugeon J, Lefevre F, Cardinal M, Uyanik A, Davenel
L. 2002. Pigmenting efficacy of astaxanthin and A. Haffray P. 2010. Flesh quality in large rainbow trout
canthaxanthin in fresh-water reared Atlantic salmon, with high or low fillet yield.
Salmo salar. Animal Feed Science Technology. 99
Bureau DP, Kaushik SJ, Cho CY. 2002. Bioenergetics.
(1/4):97-106.
In: Fish Nutrition (Eds: Halver, J.E, and Hardy, R.W)

Aba et al., 2012
Academic Press, 1-59. protine /nergie des rgimes alimentaires chez la truite
et le bar en levage .Perspectives de contrle nutritionnel
Caballero MJ, Obach A, Rosenlund G, Montero D,
des dpts lipidiques.
Gisvold M, Izquierdo MS. 2002. Impact of different
dietary lipid sources on growth, lipid digestibility, tissue Einen O, Waagan B, Thomassen MS. 1998. Starvation
fatty acid composition and histology of rainbow trout, prior to slaughter in Atlantic salmon (Salmo salar): I.
Oncorhynchus mykiss. Aquaculture 214(1-4):253-271. Effects on weight loss, body shape, slaughter- and fillet-
yield, proximate and fatty acid composition. Aquaculture
Chaiyapechara S, Casten MT, Hardy RW, Dong FM.
166(1-2),85-104.
2003. Fish performance, fillet characteristics,and health
assessment index of rainbow trout (Oncorhynchus Folch J, Lees M, Sloane-Stanley GH. 1957. A simple
mykiss) fed diets containing adequate and high method for the isolation and purification of total lipids
concentrations of lipid and vitamin E.Aquaculture, from animals tissues. J. Biol. Chem., (226):497-
219:715-738. 509.completer.
Cho CY, Bureau DP. 2001. A review of diet Gatta PP, Pirini M, Testi S, Vignola G, Monetti PG ,
formulation strategies and feeding systems to reduce 2000. The influence of different levels of dietary vitamin
E on sea bass Dicentrarchus labrax flesh quality.
excretory and feed wastes in aquaculture. Aquaculture
Aquaculture Nutrition. 6:4752.
research, 32:349-360,
Gelineau A, Corraze G, Boujard T, Larroquet L,
Choubert G. 1999. Nutrition et alimentations des Kaushik SJ. 2001. Relation between dietary lipid level
poissons et crustacs, Carotnodes et pigmentation 229- and voluntary feed intake, growth, nutrient gain, lipid
244. deposition and hepatic lipogenesis in rainbow trout.
Reproduction Nutrition Development 41:487-503.
Choubert G, Baccaunaud M. 2006. Colour changes of
fillets of rainbow trout (Oncorhynchus mykiss W.) fed Guillaume J, Medale F. 2001. Nutrition et
astaxanthin or canthaxanthin during storage under alimentations des poissons et crustacs, Nutrition
controlled or modified atmosphere Food Science and nergtique 87-111.
Technology 39(10)1203-1213.
Guroy D. 2006. Influence of Feeding Frequency on Feed
Corraze G, Kaushik S. 1999. Lipids from marine and Intake, Growth Performance and Nutrient Utilization in
freshwater fish. Ocl-Oleagineux Corps Gras Lipides European Sea Bass (Dicentrarchus labrax) Fed Pelleted
6:111-115. or Extruded Diets. Turkish Journal of Veterinary and
Animal sciences, 30,171-177.
De Francesco M, Parisi G, Medale F, Lupi P, Kaushik
S, Poli BM. 2004. Effect of long-term feeding with a Kaushik SJ, Oliva-Teles A. 1985. Effect of digestible
plant protein mixture based diet on growth and body/ energy on nitrogen and energy balance in rainbow trout.
fillet quality traits of large rainbow trout (Oncorhynchus Aquaculture 50:89-101.
mykiss). Aquaculture
Kaushik SJ, Medale F, Fauconneau B, Blanc D. 1989.
236 : 413-29.
Effect of digestible carbohydrates. On protein/energy
Dias J, Alvarez MJ, Diez A, Arzel J, Corraze G, utilization and on glucose metabolism in iainbow trout
Bautista JM, Kaushik SJ. 1998. Effet du rapport (Salmo gairdneri R.). Aquaculture 79:63-74.
Aba et al., 2012
Kaushik SJ. 1990. Nutrition et alimentation des Medale F. 2009. Les sources protiques
poissons et contrle des dchets piscicoles. La dans les aliments pour les poissons dlevage. Cahiers
pisciculture franaise, 101:14-23. Agricultures, 18(2-3):103-11
Kaushik SI. 1997. Nutrition-Alimentation et Mdale F. 2010. Les lipides des poissons daquaculture
composition corporelle chez le poisson. Cah. Nutr. Diet., et leurs facteurs de variation OCL VOL. 17 N 4
32:100-106. JUILLET-AOT 2010
Kaushik SJ. 1998. Nutritional bioenergetics and Medale F. 2010. Pratiques dlevage et qualit
estimation of waste production in non salmonids. nutritionnelle des lipides des poissons Cahiers de
Aquatic Living Resources, 11:221-217. Nutrition et de Dittique 45(5):267-273.
Kaushik SJ. 1999. Nutrition glucidique: intrt et Nickell DC, Bromage NR. 1998. The effect of dietay
limites des apports de glucides In : Nutrition et lipid level on variation of flesh pigmentation in rainbow
alimentation des poissons et crustacs INRA Editions, trout (Oncorhynchus mykiss). Aquaculture 161:237-251.
171-186.
No HK, Storebakken T, 1992. Pigmentation of rainbow
Kaushik SJ. 2000. Feed formulation, diet development trout with astaxanthin and canthaxanthin in freshwater
and saltwater. Aquaculture 101, 123-143.
and feed technology CIHEAM, 43-51.
Pfeffer E, Beckman-Toussaint J, Henrichfreise B,
Knockaert C. 2006. Salmonids daquaculture , de la
Jansen HD. 1991. Effect of extrusion on efficiency of
production la consummation. Editions Quae.327.
utilisation of maize starch by rainbow trout
Kolditz C-I. 2008. Dterminisme nutritionnel et (Oncorhynchus mykiss). Aquaculture, 96:293-303.
gntique de la teneur en lipides musculaires chez la

Pokniak. 1999. Effects of pelletization or extrusion of
truite arc-en-ciel (Oncorhynchus mykiss) : Etude par
the fattening diet on production performance of pan size
analyse de l'expression de gnes candidats, du protome
rainbow trout (Oncorhynchus mykiss) Arch. Med. vet. 31
et du transcriptome du foie et du muscle. Thse,
(1):141-150.
Universit de Bordeaux 1, Bordeaux, 420
Regost C, Arzel J, Cardinal M, Laroche M, Kaushik
Lee SM, Kim KD. 2001. Effects of dietaryprotein and
SJ. 2001. Fat deposition and flesh quality in seawater
energy levems on this growth, protein utilizationand
reared, triploid brown trout (Salmo trutta) as affected by
body compositionof juvenile Masu salmon
dietary fat levels and starvation. Aquaculture 193(3-
(Oncorhynchus masou) Brevoort. Aquaculture Research, 4):325-345.
32:39-45.
Ricker WE. 1979. Growth rates and models. In:
Liu Y Roof S, Ye Z, Barry C, van Tuinen A, W.S.Hoar, D.J. Randall and J.R. Brett (Eds.), Fish
Vrebalov J, Bowler C and Giovannoni J. 2004. Physiology Vol. VIII: Bioenergetics and Growth
Manipulation of light signal transduction as a means of
Academic Press, New York: 677-743.
modifying fruit nutritional quality in tomato. Proc Natl
Acad Sci USA, Vol.101, No.26, (June 2004) 9897-9902. Quillet E, Guillou S, Aubin J, Labb L, Fauconneau
B, Mdale F. 2007. Response of a lean muscle and a fat
muscle rainbow trout (Oncorhynchus mykiss) line on

Aba et al., 2012
growth, nutrient utilization, body composition and Torstensen BE, Lie , Hamre K. 2001. A factorial
carcass traits when fed two different diets. Aquaculture experimental design for investigation of effects of dietary
269:220-231. lipid content and pro- and antioxidants on lipid
composition in Atlantic salmon (Salmo salar) tissues and
R Development Core Team. 2011. R: A language and
lipoproteins. Aquaculture Nutrition, 7:265-276.
environment for statistical computing. R Foundation for
Statistical Computing. Watanabe T. 1982. Lipid nutrition in fish. Comparative
Biochemistry and Physiology, 73:3-15.
Richard N, Kaushik SJ, Larroquet L, Panserat S,
Corraze G. 2006. Replacing dietary fish oil by vegetable Zoccarato I, Sicuro B, Gasco L, Palmegiano GB,
oils has little effect on lipogenesis, lipid transport and Boccignone M, Bianchini ML. 1996. Use of pelletised
tissue lipid uptake in rainbow trout (Oncorhynchus or extruded diet at two feeding levels: effect on
mykiss). Br. J. Nutr., 96:200-309. performance and body composition in rainbow trout
(Oncorhynchus mykiss). Rivista Italiana Acquacoltura,
Silas SO, Hung, Trono S. 1994. Carbohydrate
31:107-118.
utilization by Rainbow Trout Is Affected by Feeding
Strategy. Journal of nutrition 124:223-230.
Silver GR, Higgs DA, Dosanjh BS, Mckeown BA,

Deacon G, Frenche D. 1993. Effect of dietary protein to
lipid ratio on growth and chemical composition of
Chinook salmon (Oncorhynchus tshawytscha) in seal
water. Ln: KAUSHIK S. J, Luquet P. (Eds.) Fish
nutrition in practice, Biarritz (France), June 24-27, 1991.
Les Colloques, No. 61. INRA, Paris, 459-468.
Song LP, AN L, Zhu, YA, Li, X, Wang Y. 2009.

Effects of dietary lipids on growth and feed utilization of
jade perch, Scortum barcoo. Journal of the World
Aquaculture Society, 40(2):266-273.
Spannhof L, Plantikow H. 1983. Studies on

carbohydrate digestion in rainbow trout. Aquaculture,
30:95-108. Submit your articles online at Ficuspublishers.com
Advantages
Torrissen O. 1986. Pigmentation of salmonids - a Easy online submission
comparison of astaxanthin and canthaxanthin as pigment Complete Peer review
Affordable Charges
sources for rainbow trout. Aquaculture 53:271-278. Quick processing
Extensive indexing
Torrissen O. 1989. Pigmentation of salmonids: Open Access and Quick spreading
interactions of astaxanthin and canthaxanthin on pigment You retains your copyright
deposition in rainbow trout. Aquaculture 79:363-374. submit@ficuspublishers.com
www.ficuspublishers.com/submit1.php.

Correlation Analysis and Exceedence Factor among the Ambient Gaseous

Pollutants and Particulate Matter in Urban Area
Authors: ABSTRACT:
Sarala Thambavani D1, Urban air pollution is rapidly becoming an environmental problem of public
Saravana Kumar R2. concern worldwide. It can influence public health and local or regional weather and
climate. In the present study, an effort has been made to study the importance of
gaseous pollutants with particulate matter. All the data were collected for a period of
eight years (2002-2009) at three different locations (Fenner (I) limited, Highway
building and Kunnathur chatram) representing industrial, residential and commercial
cum traffic areas in Madurai city. The particulate pollutants SPM and RSPM
Institution:
1. Department of Chemistry, concentrations were compared with gaseous pollutants SO 2 and NOx concentrations.
Sri Meenakshi Govt. Arts These particulate pollutants were highly compared with gaseous pollutants in all the
College for women, sampling stations during the study period. It showed significant positive correlations
Madurai- 625 002. Tamil with gaseous pollutants. The trends of pollutant levels follow the order of
Nadu, India. SO2<NOx<SPM<RSPM. The Exceedence factor was calculated for the pollutants at
different sampling areas. It reveals that Madurai city is critically polluted by RSPM and
2. Department of Chemistry, SPM at residential and commercial cum traffic area. From these values, it is clear that
N.P.R. College of the air quality is good at Fenner (I) Limited whereas it is low polluted and moderately
Engineering and polluted at Highway building and Kunnathur chatram respectively. The values of
Technology, correlation co-efficient for all possible correlations among different pollutants are
Natham, Dindigul- computed (range of r = -0.043 to 0.973). Highly significant correlation and linear
624401.TamilNadu, India. relationship are obtained between the following pairs of pollutants SO 2-NOx and NOx-
RSPM at Fenner (I) Limited, SPM-RSPM at Highway building and Kunnathur chatram
respectively.
Keywords:
Corresponding author: Ambient air quality, Exceedence factor, Oxides of Sulphur, Oxides of
Saravana Kumar. Nitrogen, SPM and RSPM, Particulate Matter.

gsivasaravanan@gmail.com Sarala Thambavani D, Saravana Kumar R.
Correlation Analysis and Exceedence Factor among the Ambient Gaseous Pollutants
and Particulate Matter in Urban Area.
Phone No: Journal of research in Biology (2012) 3: 232-240
99423-68797.
Dates:
Web Address: Received: 08 Mar 2012 /Accepted: 22 Mar 2012 /Published: 16 Apr 2012
Ficus Publishers.
cited.
232-240 | JRB | 2012 | Vol 2 | No 3

Thambavani and Kumar, 2012
INTRODUCTION hence atmospheric pollution is often severe in cities of

The origin of urban air pollution is mainly in developing countries all over the world (Mage et al.,
anthropogenic emission sources, which include 1996). Atmospheric particulate matter is the major air
automobiles, industries, and domestic fuel combustion. pollutant in India. At the same time, other chronic non-
The air pollutants so generated are detrimental to human communicable diseases such as cancer, cardiovascular
health. In addition, they cause negative impacts directly disease and respiratory disorders are becoming more
or indirectly, if at elevated concentrations, on vegetation, dominant. Approximately 50,000 premature deaths occur
animal life, buildings and monuments, weather and annually due to PM10 pollution in India. In many Indian
climate, and on the aesthetic quality of the environment cities, the levels of particulate pollutants in the ambient
(Stern, 1976; Godish, 1985; Takemura et al., 2007, Shen air have been found to be above the permissible limit
et al., 2009). The size of particulates is known to cause (Meenakshi and Saseetharan, 2004). Air quality in
severe damage to the lungs. Finer solid particles and Madurai is deteriorating. In the present study, the levels
liquid droplets, collectively called suspended particulate of particulate air pollutants were measured at three
matters, are present in the air in great numbers and at stations in Madurai city and these particulate pollutants
times they give rise to a serious pollution problem concentrations were statistically compared with gaseous
(Thambavani et al., 2012) In fact, the World Health pollutants. The present study area of Madurai city in
Organization (WHO) reports that there is no safe level Tamil Nadu consisting of large mixed use areas, having
for particulate matter emissions (WHO, 1999). high traffic volume and also fast growing center for
International studies have confirmed association between small scale industries and it was undertaken with
elevated levels of particulate air pollution and increase in following specific objective:
respiratory symptoms such as cough, shortness of breath, To conduct air quality measurement at different high
wheezing and asthma attacks. Rapid industrialization and traffic volume location of Madurai city along with
addition of the toxic substances to the environment are residential areas.
responsible for altering the ecosystem (Thambavani et To study the most common air pollutants like sulphur
al., 2011). Studies have also found associations between dioxide, oxides of nitrogen, suspended particulate matter
particulate air pollution and rates of hospitalization, and respirable particulate matter in different locations of
chronic obstructive pulmonary disease and restricted Madurai city.
activity due to illness (Dockery et al, 1993). To investigate the source of these pollutants and extent
Concentration of ambient air particulates has been found of their individual contribution to the pollution of
to be associated with a wide range of effects on human ambient air.
health (Dockery and Pope, 1994; Godberg, 1996;
Schwartz, 1991; Schwartz, 1994). The high MAJOR KINDS OF AIR POLLUTANTS
concentration of particulate matter in the environment Suspended Particulate Matter (SPM)
has become a problem for many countries (Elbir et al., SPM consists of different solid and liquid
2000). Air pollution can directly affect plants via leaves particles that are suspended in the atmosphere and
or indirectly via soil acidification (Steubing et al., 1989, includes soil, soot, lead, asbestos and sulphuric acid
Thambavani and Kumar 2011). Rates of increase of air droplets. Suspended particulates reduce visibility by
pollutant concentrations in developing countries such as scattering and absorbing sunlight, they corrode metals,
India are higher than those in developed countries and erode buildings and works of sculpture when the air is
humid and soils clothing and draperies. Smaller particles minimum of 420C and 210C respectively.
are inhaled into the respiratory system and can cause Madurai city has emerged as an important center
health problems. Lead and asbestos particles are for textiles and engineering industries. The sampling
especially harmful. stations were selected keeping in view the important
Oxides of Nitrogen (NOx) zone and the nature of activity. A total of three sampling
Oxides of Nitrogen were produced by the stations, consisting of industrial, residential and traffic
chemical interactions between Nitrogen and Oxygen. cum commercial were chosen for the present study. The
They consist mainly of Nitric oxide (NO), Nitrogen description of sampling station is given in table-1.
dioxide (NO2) and Nitrous oxide (N2O). Nitrogen For the present study, we used particulate and
oxides inhibit plant growth and when breathed, aggravate gaseous pollutant data collected by the Central Pollution
health problems, such as asthma. They are involved in Control Board (CPCB) at three sampling sites in
the production of photochemical smog, acid deposition, Madurai city over a period of eight years from 2002 to
global warming and they cause metal to corrode and 2009 and the observations made during the sampling
textiles to fade and deteriorate. period are listed in table-2 and figure-1 to 3.
Sulphur dioxide (SO2) Calculation of Correlation Co-Efficient and Linear
Sulphur dioxide was produced by the chemical Regression
interaction between Sulphur and Oxygen. SO2 is a Statistical software SPSS (Statistical package for
colorless, non-flammable gas with a strong irritating Social Sciences, Version 7.5) is used to compute the
odour and is emitted as a primary air pollutant. They correlation (r values) for all possible correlations among
corrode metals, damage stone and other materials, air pollutants. The software is used to calculate the
damage plants, and irritate the respiratory tracts of
human and other animals.
Data And Analysis Techniques
Description of study area
Madurai is one of the important cities in South
India. It is the seventh largest city in Tamil Nadu,
situated at the banks of river Vaigai and its terrain is
mostly flat. The ground rises from the city, towards
outward, on all sides except the south, which is a Figure 1: Ambient Air Quality Data at
gradually sloping terrain. It is surrounded on the outskirts Fenner (I) Limited
by small and prominent hills. The city is about 100
Table 1: Sampling sites in Madurai city
meters above mean sea level and it is situated on 9055 Location
S. No. Location Category
NL and 7807 EL and the city is covering 51.96 sq.kms description
1 Fenner (I) Industrial, Industrial
that comprises of a total population of 25,78,201 persons limited residential area
(Census 2011) whereas the Madurai Urban 2 Highway Less traffic, Residential
building residential area
agglomeration comprising the city and surrounding
3 Kunnathur High traffic, Commercial
settlements accommodates a population of 12,03,095 chatram residential, cum traffic
commercial area
persons. The climate of Madurai town is hot and dry and
and shopping
the temperature range between a maximum and complex
Table 2: Ambient air quality data of Madurai city from 2002-2009.

Avg Avg Avg
AvgSO2 PI PI PI PI
Year Location Type NOx SPM RSPM
(mg/m3) SO2 NOx SPM RSPM
(mg/m3) (mg/m3) (mg/m3)
Fenner (I)
I 18 0.22 28 0.35 65 0.18 117 0.98
Limited
Highway
R 10 0.17 26 0.43 214 1.52 137 2.29
Building
2002
Kunnathur
C&T 9 0.15 40 0.66 228 1.63 148 2.47
Chatram
Fenner (I)
I 20 0.25 31 0.39 106 0.29 98 0.65
Limited
Highway
R 12 0.20 21 0.35 138 0.98 78 1.30
Building
2003
Kunnathur
C&T 11 0.18 20 0.33 389 2.77 183 3.05
Chatram
Fenner (I)
I 19 0.24 25 0.31 144 0.40 52 0.43
Limited
Highway
R 10 0.17 20 0.33 110 0.78 50 0.83
Building
2004
Kunnathur
C&T 10 0.17 24 0.40 397 2.80 180 3.0
Chatram
Fenner (I)
I 18 0.22 26 0.32 87 0.24 29 0.24
Limited
Highway
R 10 0.17 25 0.41 104 0.74 35 0.58
Building
2005
Kunnathur
C&T 10 0.17 24 0.40 219 1.56 57 0.95
Chatram
Fenner (I)
I 13 0.16 28 0.35 113 0.31 42 0.35
Limited
Highway
R 10 0.17 26 0.43 106 0.76 37 0.62
Building
2006
Kunnathur
C&T 10 0.17 27 0.45 126 0.90 34 0.57
Chatram
Fenner (I)
I 11 0.14 24 0.30 95 0.26 40 0.33
Limited
Highway
R 8 0.13 21 0.35 93 0.66 41 0.68
Building
2007
Kunnathur
C&T 9 0.15 20 0.33 102 0.73 44 0.73
Chatram
Fenner (I)
I 11 0.14 24 0.30 89 0.24 43 0.36
Limited
Highway
R 9 0.15 22 0.37 82 0.59 38 0.63
Building
2008
Kunnathur
C&T 10 0.17 24 0.40 92 0.66 44 0.73
Chatram
Fenner (I)
I 11 0.14 26 0.32 83 0.11 41 0.34
Limited
Highway
R 10 0.17 25 0.42 85 0.61 40 0.67
Building
2009
Kunnathur
C&T 11 0.18 25 0.42 91 0.65 44 0.73
Chatram
I-Industrial area, R- Residential area, C&T- Commercial cum traffic area, PI- Pollution indices.
regression parameters A and B of the straight line Y = A sites and regression co-efficient values (A and B) are
+ BX by applying the well-known method of least tabulated in tables-3, 4, 5, 6, 7 and 8.
squares (Wonnacott and Wonnacott, 1981, Gupta, 1974) Exceedence Factor
to fit the experimental data to give straight line. The The air quality has been categorized into four
correlation co-efficient (r) values at different sampling broad categories based on an Exceedence Factor (EF).
Figure 2: Ambient Air Quality Data at Figure 3: Ambient Air Quality data at
Highway Building Kunnathur Chatram
The four air quality categories for Exceedence Factor Madurai city at three sites, namely industrial, residential
values are given in table-9. It is calculated as follows: and commercial cum traffic areas were considered to
determine the short-term air quality indices. Annual
Observed annual mean concentration of criteria pollutant
Exceedence Factor = average values of critical parameters SPM, RSPM, SO2
Annual standard for the respective pollutant
and NOx, which generally persist in the air under the
It could be seen from the above categorization, prevailing microclimate conditions, are considered.
that the locations in either of the first two categories are Any correlation will be statistically significant
violating the standards, although, with varying only if its r values are very close to +1 or 1. In the
magnitude. Those, falling in the third category are present investigation, tables-3, 4 and 5 show the
meeting the standards but likely to violate the standards correlation co-efficient of pollutants. High positive
in future if pollution continues to increase and is not correlations were observed between SPM and RSPM
controlled. However, the locations in low pollution (0.97) at Highway Building, and Kunnathur chatram
category have a rather pristine air quality and such areas (0.92) and NOx-RSPM (0.68) at Fenner (I) Limited
are to be maintained at low pollution level by the way of whereas negative correlation were found between NOx
adopting preventive and control measures of air and SPM (-0.042), SPM and RSPM (-0.26) at Fenner (I)
pollution. Computed Exceedence factor categories for Limited, SO2 and NOx (-0.45) and NOx-SPM (-0.055) at
various pollutants at three sampling areas are tabulated in kunnathur chatram respectively. Tables 6, 7 and 8 show
table-10. the regression co-efficient for all the possible pairs of
pollutants.
RESULTS AND DISCUSSION Pairs having very high positive correlation
In the present study, the annual average value of between them show the dependency of one pollutant on
SO2, NOx, SPM and RSPM in the ambient air of
Table 4: Correlation co-efficient (r) between gaseous
Table 3: Correlation co-efficient (r) between pollutants and particulate matter at Highway
gaseous pollutants and particulate matter at Building
Fenner (I) Limited
Parameters (mg/m3) Parameters (mg/m3)
Parameter Parameter
SO2 NOx SPM RSPM SO2 NOx SPM RSPM
SO2 1 0.063618 0.366364 0.335073
SO2 1 0.57815 0.25793 0.563495 NOx - 1 0.30704 0.214817
NOx - 1 -0.04288 0.675964
SPM - - 1 0.973793
SPM - - 1 -0.26226
RSPM - - - 1 RSPM - - - 1
Table 5: Correlation co-efficient (r) between gaseous Table 7: Regression coefficient (A and B) between
pollutants and particulate matter at Kunnathur gaseous pollutants and particulate matter at
chatram Highway Building.
Parameters (mg/m3) Parameter pairs A B
Parameter
SO2 NOx SPM RSPM NOx and SO2 0.140845 21.85915
- NOx and SPM 0.017753 21.18179
SO2 1 0.222074 0.09994
0.44821 NOx and RSPM 0.015207 22.38318
NOx - 1 -0.05539 0.172352 SPM and SO2 14.02817 -22.0282
SPM - - 1 0.927693 SPM and NOx 5.310345 -6.96552
RSPM - - - 1 SPM and RSPM 1.192281 48.53998
RSPM and SO2 10.47887 -46.4789
the other while pairs having negative correlation among RSPM and NOx 3.034483 -13.5517
RSPM and SPM 0.795343 -35.6574
them suggest increase in concentration of one pollutant
represents decrease in other. The values of regression co- annual average concentration of 65 mg/m3 in the year of
efficient (A and B) greatly help in finding out the 2002. But the level of pollutant SPM are within the
regression equation between the two pollutants, which prescribed limit throughout the year. The level of RSPM
can be further extended in 95 % confidence limit. exceeded the prescribed limit during the year 2002
In the present study at Madurai city, primary to2004 at the sampling station of Kunnathur chatram.
pollutants such as SO2 and NOx were found within the The values of SPM are 148,183and180 mg/m3
prescribed limit and SPM and RSPM were found to be respectively. The level of pollutant sulphur dioxide and
exceeding the prescribed limit. That is the annual mean oxides of nitrogen were found within the prescribed limit
SPM level at all the three sites remained above Indian of CPCB throughout the years in all the study area.
National Ambient Air Quality Standard. The annual Fenner (I) Limited
average values of SPM, SO2, RSPM and NOx in the In the Fenner (I) Limited, the level of SPM is
ambient air of Madurai city are considered to determine highest and SO2 is lowest. The trend of pollutants levels
the pollution trends. This shows that maximum SPM is SO2<NOx<RPM<SPM. It is found that the levels of
levels were found in Kunnathur chatram with annual SPM and RSPM have exceeded the standards in all the
average concentrations of 397 and 389 mg/m3 during the years. The correlation analysis reveals that there is no
year 2004 and 2003 respectively, which is observed three high positive correlation between the pollutants except
times above the standard value. The low concentration of correlation values of 0.57 and 0.68 are observed for NOx
SPM was found at industrial site Fenner (I) limited with - SO2 and NOx - RSPM respectively.
Table 6: Regression coefficient (A and B) between Table 8: Regression coefficients (A and B) between
gaseous pollutants and particulate matter at gaseous pollutants and particulate matter at
Fenner (I) Limited. Kunnathur chatram.
Parameter pairs A B
Parameter pairs A B
NOx and SO2 0.347238 21.24803
NOx and SPM -0.00433 26.92289 NOx and SO2 -3.75 63
NOx and RSPM 0.050911 23.55988 NOx and SPM -0.00274 26.06394
NOx and RSPM 0.01647 23.98886
SPM and SO2 1.535513 74.52537
SPM and SO2 37.5 -169.5
SPM and NOx -0.425 109.0125 SPM and NOx -1.11786 234.0054
SPM and RSPM -0.19577 109.0559 SPM and RSPM 1.789247 41.33658
RSPM and SO2 4.493799 -10.2187 RSPM and SO2 8.75 4.25
RSPM and NOx 8.975 -180.088 RSPM and NOx 1.803571 45.75893
RSPM and SPM -0.35132 92.09123 RSPM and SPM 0.480992 -7.09388
Table 9: Relative scale for Exceedence Factor values the study areas. In order to control the pollutant levels in
Exceedence Factor values Categories urban areas, urgent attention is required for controlling
> 1.5 Critical pollution
vehicular emission and industrial emission. Air pollution
1.0 1.5 High pollution
0.5 1.0 Moderate pollution management for urban air quality monitoring and
< 0.5 Low pollution assessment should provide management approaches by
Highway Building deriving policies and strategies.
In the Highway building, the level of SPM and In the present study, the particulate pollutants
RSPM is highest during the year 2002 and the values are SPM and RSPM are mostly above the permissible limit
214 and 137 respectively. It is found that the level of at Kunnathur chatram in Madurai city. The gaseous
pollutants are decreasing trend in all the years. The trend pollutants SO2 and NOx concentrations in Madurai city
of pollutants is SO2<NOx<RSPM<SPM. High positive are found to be decreasing trend which is in accordance
correlation exists for the following pair SPM and RSPM with Bhaskar et al., 2008, Bhaskar et al., 2010). It
in this area. implies that particulate matters are produced by vehicles,
Kunnathur Chatram industrial activities and combustion of conventional fuels
At Kunnathur chatram, the level of SPM and for domestic and commercial purposes. The results of
RSPM is higher than the level of sulphur dioxide and this study will be useful for further research on
oxides of nitrogen. The trend of pollutant levels are interactions between atmospheric aerosols, local and
SPM>RSPM>NOx>SO2. SO2 and RSPM, SPM and regional weather and climate in Madurai city.
RSPM are positively correlated with r-values 0.92 for Sustainable Development And Planning
each. This shows that RSPM concentration increases The preparation of Environmental Management Plan
with the concentrations of SO2 and SPM. (EMP) studies should be taken up for tourism and
Comparison of Exceedence Factor values of all environmentally fragile areas.
the sampling areas shows that the air quality is critically The sustainable development plan requires the
polluted by RSPM and SPM at residential and conservation of environment resources, like forest,
commercial cum traffic area for all the years. Industrial natural gases flora and fauna and qualities to protect the
sites of Madurai city are moderately polluted from 2002 interests of our future generations.
to 2009.The level of SO2 and NOx are quite clean in all The Environment Management Plans for urban areas
Table 10: Computed Exceedence factor categories for various pollutants at three sites during 2002-2009.
Sampling Periods
Sampling areas Pollutants
2002 2003 2004 2005 2006 2007 2008 2009
Fenner (I) SO2 L L L L L L L L
Limited NOx L L L L L L L L
SPM L L L L L L L L
RSPM M M L L L L L M
Highway SO2 L L L L L L L L
Building NOx L L L L L L L L
SPM C M M M M M M M
RSPM C H M M M M M M
Kunnathur SO2 L L L L L L L L
Chatram NOx M L C L L L L L
SPM C C C C M M M M
RSPM C C C C M M M M

should be targeted to provide planning solutions for the pollution.

different kinds of urban areas wherein the environmental Green belt may be provided throughout city. It will
degradation has taken place and environmental risks are reduce excess CO2, provide health-giving oxygen, trap
increasing (MoEF, 2000-2001). aerosols, supply biomass and timber and create social
The concentrations of SPM observed in the present forestry.
study are indicative and continuous monitoring at Pollution level may be monitored regularly. Increasing
minimum five categories, that is residential, commercial, awareness among the people.
industrial, petrol pumps and traffic junctions is
recommended as this will help in anticipating future REFERENCES
concentration of SPM in the ambient air, formulating Air Quality Management. 1999. Air Quality
standard levels and decision pertaining to control air Guidelines, WHO, Geneva.
pollution (Gupta et al., 2005).
Bhaskar BV, Rajasekhar RVJ, Muthusubramanian P
Recommendation
and Kesarkar AP. 2008. Measurment and Modelling of
As per the results, there are some suggestions, which
Respirable Particulate (PM10) and Lead Pollution over
can be adapted for prevention, abatement and control of
Madurai, India. Air Qual. Atmos. Health 1:45-55.
air pollution. It may be understood that these measures
are suitable to be applied at the source itself, since the Bhaskar B Vijay, Vikram M. Mehta. 2010.
pollutants are released in the atmosphere, it is only by Atmospheric Particulate Pollutants and their Relationship
virtue of the natural phenomena of wind, and thermal with Meteorology in Ahmedabad. Aerosol and Air
convection and rain wash out that their extent can be Quality Research. 10:301-315.
reduced.
Dockery DW. 1993. New Engl. J. Med., 329:1753-1759.
Vehicular pollution can be reduced or controlled
through the implementation of Euro/Bharat norms and Dockery Ward D, Pope CA. 1994. Acute respiratory
phase out of old and ill maintained vehicles. effects of particulate air pollution, Annu. Rev. Public
CNG may be introduced in lieu of petrol. Health, 15:104-132.
Traffic rules should be implemented properly avoiding Elbir T, Muezzinoglu A and Bayram A. 2000.
traffic jams, smooth traffic flow and it should be Evaluation of some air pollution indicators in Turkey,
regularized by imposing nominal pollution tax for those Environmental International, 26:5-10.
who float the city traffic rules and regulations.
Polluting industries should be away from the city. They Godberg K. 1996. Particulate air pollution and daily
should have one complex, where in all the industrial mortality: Who is at risk? Journal of Aerosols Med., 9:
pollution norms are observed. Offices should not be 43-53.
concentrated in silence zone. Godish T. 1985. Air Quality. Lewis Publishers, Inc.
Road should be well maintained. Bye-pass roads may Chelsea, USA.
be provided for inter city traffic. Flyover may be built to
Gupta HK. 2005. Analysis of Dioxin in the
avoid traffic jams.
environment. Indian J. Env. Prot., 25(2):159-167.
Common diesel generator sets should be established in
the light cut hours to avoid unnecessary air and sound

Gupta SP. 1974. Advanced practical statistics, 2nd 39-60.

edition, S.Chand, New Delhi.
Shen ZX, Cao JJ, Tong Z, Liu SX, Reddy LSS, Han
http:/www.cpcb.delhi.nic.in/as.htm. 2004. Central YM, Zhang T and Zhou J. 2009. Chemical
Pollution Control Board, New Delhi. Characteristics of Submicron Particles in Winter in
Xian. Aerosol Air Qual. Res. 9: 8093.
http:/homeiitk.ac.in/-mukesh/basis.html. 2004. Basis
for Indian Air Quality Index (INDAQI), Stern AC. 1976. Air Pollution. 1, 3rd edn. Academic
Press, New York, USA.
Mage D. Ozolins G, Peterson P, Webster A, Orthofer
R, Vandeweerd V, Gwynne M. 1996. Urban air Steubing L, Fangmier A. 1989. Both, R; Effects of
pollution in mega-cities of the world. Atmospheric SO2, NO2, and O3 on Population Development and
Environment; 30:681-686. Morphological and Physiological parameters of Native
Herb Layer Species in a Beech Forest. Environmental
Meenakshi P and Saseetharan MK. 2004. Urban Air
Pollution 58:281-302.
Pollution Forecasting with Respect to SPM using Time
Series Neural Networks Modelling Approach A Case Takemura T, Kaufman YJ, Remer LA, Nakajima T.
Study in Coimbatore City. J. Environ. Sci. Eng., 46:92- 2007. Two competing pathways of aerosol effects on
101. cloud and precipitation formation, Geophys. Res. Lett.
34: L04802. doi: 10.1029/2006GL028349.
MoEF. 2000-2001. Annual report. Ministry of
Environment and Forests, Government of India, New Wonnacott TH and Wonnacott RJ. 1981. A second
Delhi. course in statistics, John Wiley and sons, New York.
SaralaThambavani D and Saravanakumar R. 2011.

Effect of Cement dust on Photosynthetic pigments of
selected plant species Asian J.Environ.Sci., 6(2):161-
167.
Sarala thambavani D, Saravana Kumar R. 2011. The

monthly changes of chloroplast pigments content in
selected plant species exposed to cement dust pollution.
Journal of research in Biology. 8: 660-666.
Sarala thambavani D, Vidya vathana M. 2012. Submit your articles online at Ficuspublishers.com
Ambient concentration of suspended particulate matter Advantages

and manganese in urban area of Madurai City. Journal of Complete Peer review
research in Biology. 1: 001-006. Affordable Charges
Quick processing
Schwartz J. 1991. Particulate air pollution and daily Extensive indexing
mortality: A Synthesis Public health Rev., 19:39-60.
Schwartz J. 1994. Air pollution and daily mortality: A submit@ficuspublishers.com

review and metal analysis. Environmental Research, 64:

Ichthyodiversity of the Rangavali Dam, Navapur,

District Nandurbar, Maharashtra State.
Authors: ABSTRACT:
Jaiswal DP and
Ahirrao KD. The study area was Rangavali dam of Navapur, district Nandurbar
(Maharashtra State) India. This dam has not received much attention by limnologist
and this prompted us to sample the fishes throughout the year to assess the
Institution: Icthyodiversity. Fishes were collected from the catch of local fisherman at different
A.S.C. College, Navapur,
stations of dam, from June 2007 to May 2009 on a monthly basis. Initially fishes were
Dist-Nandurbar (MS), India.
identified by local name and common name as named by local fishermen and then the
Rani Laxmibai College, scientific identification and classification were made. In the present study, 28 fish
Parola, Dist-Jalgaon, species were found, belonging to 25 genera and 12 families were grouped under
425111 India. seven orders. Among 28 fish species the order Cypriniformes was found to be
dominant.

Ahirrao KD. Freshwater Rangavali Dam, Ichthyodiversity.

drkiranahirrao@gmail.com. Jaiswal DP and Ahirrao KD.
Ichthyodiversity of the Rangavali Dam, Navapur, District Nandurbar, Maharashtra
State.
Phone No: Journal of research in Biology (2012) 3: 241-245
+919455784578
Dates:
Received: 30 Jan 2012 /Accepted: 06 Feb 2012 /Published: 19 Apr 2012
Web Address:
Documents/RA0194.pdf. Ficus Publishers.
cited.
241-245 | JRB | 2012 | Vol 2 | No 3

Jaiswal and Ahirrao, 2012
INTRODUCTION freshwater fishes are useful to examine factors

The entire study area is declared as tribal area by influencing the structure of the fish community. The
the Central Government of India. The country is distribution and composition of the fish species in each
endowed with vast and varied water resources and rich habitat were closely associated with various factors such
biodiversity. Freshwater fishery sites are varied; like as the availability of food, breeding sites, water current,
45,000 Km of rivers, 1,26,334 Km. of canals, ponds and depth, topography and physico-chemical properties of
tanks 2.36 million hectares of reservoirs. The fish fauna water (Harris, 1995).
is divided into two classes, viz., Chondrichthyes Biodiversity may be broadly defines as the
(cartilagenous fishes) and Osteichthyes (bony fishes). variety and variability among living organisms and the
The endemic fish families form 2.21 percent of the total ecological complexes in which they occur. Biodiversity
bony fish families of the Indian region. 223 endemic fish can be considered at different scales ranging from the
species are found in India, representing 8.75 percent of gene to ecosystem. The most commonly used meaning of
the total fish species known from the Indian region. biodiversity is at the level of species (Organismal
There are about 450 families of freshwater fishes biodiversity). Freshwater systems harbor a unique and
globally. Roughly 40 are represented in India. About 25 diverse set of organisms. About 15 % of all animal
of these families contain commercially important species that have been described until today live in
species. Number of endemic species in warm water is freshwater ecosystems. More than 70 thousand
about 544. Freshwater fishes are a poorly studied group freshwater species from 570 families and 16 phyla have
since information regarding distribution, population been describing so far (Strayer 2001).
dynamics and threats are incomplete, and most of the
information available is from a few well-studied MATERIALS AND METHOD
locations only. Biodiversity is essential for the This study was conducted at in the Rangavali
stabilization of ecosystem, protection of overall dam. Samples were collected from various stations
environmental quality, for understanding intrinsic worth between June 2007 and May 2009 on a monthly basis,
of all species on the earth. In India, there are 2,500 using 1 to 2 mm pore size fish net, and with horizontal
species of freshwater fishes that have been recognized in and vertical hauls. The samples were evaluated
the Indian subcontinent out of which 930 are categorized quantitatively, and the species were identified from
as freshwater species by Day,(1878) Jayaram(1981), collected samples. Initially fishes were identified by
Talwar and Jhingran (1991) and Rao et al.,(1999), local name and common name as named by local
Sahare and Joshi(2002), Dutta et al.,(2003), Sakhare and fishermen. The scientific identification and classification
Joshi(2004), Yadav (2005), Battul et al.,(2007), were made by using the keys of Day (1878). Later
Ashashree et al ., (2008). In present study an attempt has specimens were preserved in 4 % formaldehyde.
been made to highlight the Ichthyofaunal diversity of Topography and Morphometry:
Rangavali Dam. The work will provide future strategies Rangavali Dam is known as Rangavali river
for betterment of nearby village tribals because many of project in government documents. In the year 1972, it is
the villages are surrounded to the said dam are built over Rangavali river near Nagziri village, taluka
exclusively from tribal category. Navapur, district Nandurbar, Maharashtra. The average
Studies of spatial and temporal pattern of annual rain fall in the surrounding area is 1227 mm, the
diversity, distribution and species composition of catchment area of dam is 99.20 Sq. Km. The geographic
location of the dam is 73 0 52 0 longitudinal and 210 0 dominant by consisting ten species while order
latitude. The gross capacity of the dam is about 15.02 Mugiligormis and Synnbranchiforms consists single
Mcum and capacity of the dead storage 2.13 Mcum Dam species only.
is earthen of rolled filled maximum height of the dam in Table 1: List of fishes recorded from Rangavali
Dam-for 2007-08 and 2008-09.
the river is 25.63 meter and the length is 1878 meter. The Sr. No. Classification
earthwork of the dam is 1.042 Mcum, concrete 113.00 Class: Pisces
Sub-Class Teleostei
Mcum, masonry 7770 Mcum and excavation 327134 Order-I: Clupeiformes
Mcum. The entire area is declared as a tribal area by the Family- I: Notopteridae
1 Notopterus notopterus
central government of India. This dam has not received 2 Notopterus chital
much attention by limnologist and this prompted us to Order-II: Cypriniformes
Family-II: Balitoridae
sample the fishes throughout the year to assess the
3 Nemacheilus moreh
diversity of fishes. Family-III: Cyprinidae
4 Labeo rohita
5 Cyprinus carpio
RESULTS 6 Rasbora daniconias
In the present study, 28 fish species were found. 7 Puntius sophore
8 Thynicthys sandkhol
Among 28 fish species, 25 genera and 12 families were 9 Salmostoma novacula
grouped under seven orders. 10 Catla catla
11 Cirrhinus mirgala
1. Order Clupeiformes contained a two species namely
12 Garra mullya
Notopterus notopterus and Notopterus Chitla. Order-III: Siluriformes
Family-IV: Bagridae
2. Order Cypriniformes contained 10 species namely
13 Mystus aor
Nemacheilus moreh, Catla catla, Cirrhinus mrigala, 14 Mystus seenghala
Labeo rohita, Cyprinus carpio, Rasbora daniconias, 15 Rita rita
Family-V: Siluridae
Puntius sophore, Thynicthys sandkhol, Salmostona 16 Wallago attu
novacula and Garra mullya. 17 Ompak bimaculatus
Order -IV: Mugiliformis
3. Order Siluriformes contained five species namely Family-VI:Mugilidae
Mystus aor, Mystus seenghala, Wallago attu, Ompak 18 Rhinomugil carsula
Order-V: Synnbranchiformes
bimaculatus and Rita rita.
Family-VII: Mastacembelus
4. Order Channiformes contained four species namely 19 Mastacembelus armatus
Clarius batrachus, Channa gachua, Channa Order-VI: Channiformes
Family-VIII: Channidae
marulius and Channa punctatus. 20 Clarius batrachus
5. Order Synnbranchiforms contained a single species 21 Channa gachua
22 Channa marulius
namely Mastacembelus armatus. 23 Channa punctatus
6. Order Perciformes contained a five species namely Order-VII: Perciformes
Family IX Chandidae
Oriochromis mossambicus, Chanda nama,
24 Chanda nama
Glossogobius giuris giuris, Xentodon cancilla and 25 Parambassis ranga
Family X Cichlidae
Parambassis ranga.
26 Oreochromis mossambicus
7. Order Mugiliformis consisted the only one species Family XI Gobidae
Rhinomugly carsula. 27 Glossogobius giuris giuris
FamilyXII Belonidae
The order Cypriniformes was found to be 28 Xenetodon cancila
DISCUSSION AND CONCLUSION Mugiliformes and Beloniformes. The order

The fishes are the major component of an aquatic Cypriniformes was dominant.
ecosystem having high economic value, as they provide The fish community in lakes include the native
the nutritious and delicious food for mankind. They also species and the introduced species for the purpose of fish
provide protein rich food and several economically production. Many of the fish species are endemic to this
important byproducts. The fish are also important to keep region. To conserve all endemic fish species and the total
ecosystem in balance and enhances the beauty of the fish diversity, it is necessary to prevent drainage of
nature in different ways. About 30,000 to 40,000 species pesticides and fertilizers from surrounding crop fields,
are reported differing widely from each other in shape, heavy siltation during heavy rainfall, high density of
size habits and habitats. Some of the fishes are very fingerling stocking selected culture fishes, fish diseases.
small measuring not more than an inch in length, while a Sustainable fish production by taking appropriate steps
few species attend a length up to 18.50 meters. In the for sustaining fish diversity is necessary to conserve
field of ichthyology valuable contribution were made by these vulnerable resources.
Rahimullah (1943), Chacko and Thyagarajan(1954),
David(1963), Das(1966), Karamchandani and Pisolkar ACKNOWLEDGEMENT
(1967) and Saha(1970) Rathod and Khedkar(2011). Authors are grateful to Dr. K. B. Shejule, Asso.
As far as Rangavali Dam (Navapur, district Professor, Dept of Zoology, Dr. Babasaheb Marathwada
Nandurbar, MS, India) is concerned poor attention has University, Aurangabad (Maharashtra) for his constant
been paid towards systematic investigation on diversity encouragement and providing the laboratory facilities
of fish fauna. So it is felt that there is a need to generate during the course of study.
information on diversity of fishes from Rangavali
reservoir. Hence, the present investigation was REFERENCES
undertaken to prepare a check list of fishes from Ahirrao SD and Mane AS. 2000. The diversity of
Rangavali Dam and it is the first effort in this direction. ichthyofauna, taxonomy and fisheries form some fresh
Babu Rao (1997) has studied the fish fauna in waters of Prabhani District (MS), J. Aqua, Biol.,15:40-
Himayatsagar lake in Hydrabad and recorded 32 fish 43.
species belonging to six order with 11 families. In recent
Ashashree HM, Venkateshwarlu M and Reneuka
years Rao et al.,(1999) has studied Icthyofauna of river
Swamy HM. 2008. Diversity of fish fauna in
Nagavalli from A. P.; Ahirrao and Mane(2000)has
Nagathibelagulu Pond, Shimoga, Karnataka. Advances in
studied Icthyofauna from Parabhani District of
aquatic ecology (2):95-97
Maharashtra State and Sakhare (2001) of Jawalgaon
reservoir in Solapur District of Maharashtra. Very Babu Rao M. 1997. Studies on the ecology and fish
recently Walujkar (2005) reported 35 Species from fauna of an Oligotrophic lake Hamayatsagar Hydrabad
Shirsathwadi reservoir, Mohari reservoir and (A.P.), Rec. Adv. In freshwater Biology II (8):123-138.
Manikdaundi reservoir of Nagar District Maharashtra
Battul PN, Rao KR, Navale RA, Bagale MB and
State. They further illustrated 18 species form order
Shah VN. 2007. Fish diversity from Errukh Lake near
Cypriniformes, five species in order Siluriformes, four
Solapur, Maharashtra. J.Aqua. Biol., 22(2):68-72.
species in Preciformes, one species each for order
Clupeiformes, Channiformes, Mastacemboeliformes,
Chacko Kurion and Thyagarajan. 1954. Survey of Sahare VB and Joshi PK. 2002. Ecology of Palas NI
fishes of Cauvery River, Freshwater fish Biol. Stn. Legaon reservoir in Osmanabad district Maharashtra. J.
Madras. 12:19. Aqua.Biol., 18(2):17-22.
Das SM. 1966. The Ichthiyofauna of Kashmir, Pro. Nat. Sahare VB and Joshi PK. 2004. Present status of
Acad. Sci. India. 33/b(2):62-69. reservoir fishery in Maharashtra. Fishing Chimes 24
(8):56-60.
David A. 1963. Studies on fish and fisheries of
Godavari and Krishna River system. Part-I Pro. Nat. Sakhare VB. 2001. Ichthyofauna of Jawalgaon reservoir
Acad.of Sci., India 33(2):263-286. in Solapur District (M.S.) J. Aqua. Bio., 16(1&2):31,33.
Day FS. 1878. The fishes of India, Williams and Sons Strayer DL. 2001. Ecology and distribution of
ltd. London. hyporheic microannelids (Oligochaeta, Aphanoneura and
Polychaeta) from the eastern United States: Archiv for
Dutta SPS, Kour H and Zutshi N. 2003. Ichthyofauna
Hydrobiology 131(3):493-510.
of river Tawi and its Tributories, and important tributary
of the river Chenab J. Aqua. Biol., 18(2):61-68. Talwar PK and Jhingran AG. 1991. Inland fishes of
India and adjacent countries Vol. I and II Oxford,
Harris JH. 1995. The use of fish in ecological
Clarendon Press (1908-1931).
assessments. Australian Journal of Ecology 20:65-80
Walujkar AG. 2005. Hydrobiology study of Dam water
Jayaram KC. 1981. The freshwater fishes of India,
from Pathardi Dist. Ahmadnagar (M.S.)Ph. D. Thesis.
Pakistan, Burma and Sri Lanka. Hand book of
Zoological survey of India. No.2 xxii+475. Yadav BE. 2005. Pisces, Fauna of Nathsagar Wetland,
Wetland Ecosystem Series. Zool. Surv. India. 7:137-143.
Karamchandani SJ and Pisolkar MD. 1967. Survey of
fish and fisheries Tapi river. Survey report Centre,
Inland fish. Res. Inst. Bharakpore. 4-29.
Rahimullah M. 1943. Fish survey of Hyderabad City

and its suburbs J. Bom. His. Soc., 44(1):88-91.
Rao A. Sreenivasa, Rao P. Rammohan and

Somesswara Rao. 1999. Degradation of Kolleru lake
Ind. J. Env, Hlth., 41(4):300-311.
Rathod SR and Khedkar GD. 2011. Impact of Advantages
elevation, longitude and longitude on fish diversity in
Godavari river Journal of Research Biology 4:269-275. Affordable Charges
Quick processing
Saha GN, Sahagal KL, Mitra E and Nandy AI. 1970. Extensive indexing
Studies on the seasonal and Diurnal variations in
physicochemical and biological conditions of a perennial
fresh water pond J. Ind. Fish Soc. India. 3:79-102. www.ficuspublishers.com/submit1.aspx.

Short communication Publication group
Prevalence of gastro-intestinal nematodiasis in Black Bengal goat of

Sylhet Govt. Goat Development Farm, Bangladesh
Authors: ABSTRACT:
Akanda1 MR, Hossain2
FMA, Uddin3 MN, Belal2
SA, Ashad4 FA, Howlader1
MMR. A survey on the prevalence of gastro-intestinal tract (GIT) parasites in 20
black Bengal goats (Capra hircus) of 18 months of age was conducted in Sylhet Govt.
Goat Development Farm, Bangladesh during the period of February to May of 2011.
Institution:
Irrespective to sex, using McMaster method for egg per gram of feces (EPG) disclose
1.Department of Physiology
and Pharmacology. that the percentage of Haemonchus contortus, Strongyloides papillosus, Trychuris ovis,
Trychostrongylus vitrinus., Oesophagostomum columbianum and mixed infections
2.Department of Dairy and were prevalent as 30, 16.67, 10, 6.67, 13.33, and 23.33 respectively. Study surveys
Poultry Science. suggest, appropriate parasitic control approach be explored and tried in order to
alleviate the problem of worm saddle.
3.Department of Animal
Nutrition and Livestock
Management; Faculty of
Veterinary and Animal
Science; Sylhet Agricultural
University, Sylhet-3100;
Bangladesh.
4. Senior Instructor,
Livestock, Youth Training
Centre, Ministry of Youth
and Sports, Bangladesh.
Corresponding author: Article Citation:

Hossain FMA. Akanda MR, Hossain FMA, Uddin MN, Belal SA, Ashad FA, Howlader MMR.
Prevalence of gastro-intestinal nematodiasis in Black Bengal goat of Sylhet Govt. Goat
Development Farm, Bangladesh.
Email:
fmhossainvet@yahoo.com Dates:
Received: 08 Jan 2012 Accepted: 15 Jan 2012 Published: 21 Apr 2012
Ficus Publishers.
Web Address: This Open Access article is governed by the Creative Commons Attribution License (http://
http://jresearchbiology.com/ creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-
documents/RA0183.pdf. commercial, distribution, and reproduction in all medium, provided the original work is properly
cited.
246-250 | JRB | 2012 | Vol 2 | No 3

An International Open Access Online Submit Your Manuscript
Akanda et al., 2012
INTRODUCTION (Perry et al., 2002; Sahlu et al., 2009). The prevalence of

Goat rearing is becoming more and more popular anthelmintic resistance is a serious constraint to goat
in Bangladesh. Parasitism has been considered as one of production globally (Howell et al., 2008; Kaplan et al.,
the major constraints of livestock production. Goat 2004). The use of sustainable, integrated parasite control
rearing contributes greatly to the poverty stricken rural systems, using scientifically proven non-chemical
people, especially to small and marginal farmers and methods and limited use of drugs is being considered to
landless laborers holding less than 2 acres of land ensure animal health and food safety (Waller, 2006).
(Husain et al., 1998; SAIC, 1995). Among the
constraints, helminthiasis especially parasitic MATERIALS AND METHODS
gastro-enteritis (PGE) constitutes a serious health This work was carried out in Government Goat
problem and limitation to the productivity of small Development Farm, Sylhet located at 24.8917N
ruminants (Goats and Sheep) throughout the world due 91.8833E, Bangladesh from February to May, 2011.
to the associated morbidity, mortality and cost of Twenty (20) goats were selected irrespective to sex and
treatment and control measures (Silvestre et al., 2000). the fresh fecal sample (near to 5 gm) collected from each
They cause the animals to be unthrifty which may black Bengal goat (C. hircus) early in the morning on
include the loss of weight, low birth weight, and weekly interval with aseptic precautions and transferred
difficulty in kidding. Parasitisms are important limiting that immediately to the laboratory of Department of
factors that are responsible for deteriorating the health Physiology and Pharmacology, Sylhet Agricultural
and productivity of livestock. Parasitic infestations exert University. All samples kept in refrigerator at 4C
adverse effects on the health and productivity of animals temperature for onward examination. Egg per gram of
(Rehman et al., 2009). These effects are varied and more feces (EPG) from the naturally infested goats was
pronounced in sheep and goats compared to those seen in monitored at weekly intervals. EPG was recorded just
other species of livestock (Iqbal et al., 1993). Many prior to treatment from each group. EPG of experimental
species of parasites are seen in sheep and goats and goat was determined by McMaster method. In this
usually include Haemonchus, Oesophagostomum, method a known volume of feces (5gm) was thoroughly
Ostertagia, Cchabertia, Nematodirus, Trichuris, suspended in a known volume of (50ml) saturated salt
Moniezia and Fasciola. The most important of these is solution. The suspension was stirred through a 150 mm
Haemonchus contortus (Husnain and Usmani, 2006). It mesh sieve to remove the course particles. A portion of
is an important blood sucking parasite of the ovines and the suspension was withdrawn with the help of Pasteur
causes an insidious drain on production (Asanji and pipette and allowed to run into the chambers of the
Williams, 1987; Ijaz, et al., 2008), weight losses and McMaster slide. The slide was allowed to stand for 5
even mortality in young animals (Husnain and Usmani, minutes to allow the eggs to float.
2006). The disease caused by various gastrointestinal The eggs in the two chambers were counted
nematodes is prevalent wherever sheep and goats are using low power objectives (10). The number of eggs
raised, but it exerts the greatest economic losses in per gram of feces was calculated by using the following
temperate and tropical regions (Blood et al., 1979; Raza formula.
et al., 2009; Ijaz et al., 2009). Number in two chambers
Number in one gm = x dilution factor
Gastrointestinal parasites pretense the greatest 0.3
challenge to goat health and production in humid areas
Akanda et al., 2012
Total volume of suspension in ml Waruiru et al., (2003).

Dilution factor =
Seasonal variations of gastro-intestinal
Total volume of feces
Weekly EPG count was also done on day 7, 14, nematodes should be considered due to their distribution
21 and 28 post treatment by McMaster egg counting and frequency of larval stages. Our findings observed
technique. from February to May and this is naturally a time
Statistical analysis between late winters to initiation of summer in
Descriptive statistics were used to analyze the Bangladesh. This finding also corroborated with Mazid
mean EPG of the helminth egg count, prevalence, and et al., (2006) and Zong et al., (1997) investigated the
pre-treatment and post-treatment results. Linear g ast ro - int est inal ne mat o d es lar vae of
correlation was used to analyze differences between Haemonchus cont ort us , Nematodi rus and
body condition scores and EPG counts, and EPG of Oesophagostomum were higher in January, February and
different body scores were analyzed by pair-wise mean December.
comparisons using SPSS v.15 for Windows (SPSS, Inc., In a column figures with same letter or without
Chicago, IL, USA). letter do not differ significantly whereas figures with
dissimilar letter differ significantly (as per DMRT)
RESULTS AND DISCUSSIONS Similarly, the prevalence of Trichuris (10.00%)
In the present study, Haemonchus, strongyloides, observed in the present study is also in conformity with
Trichuris, Oesophagostomum, Trichostrongylus and result observed by Fivaz et al., (1990) and Broomker et
mixed infestations were observed before treatment. al., (1989) in Angora goats though Samanta and Santra
Among the parasite the prevalence was highest for (2009) found Trichuris spp only as 2.15%. On the
Haemonchus (30.00%) and this was followed by mixed contrary, the prevalence of Trichostrongylus sp. (6.67%)
infestation (23.33%) strongyloides (16.67%), observed in the present study was lower than the findings
Oesophagostomum (13.33%), Thichuris (5.77%) and made by Gupta et al., (1987). The prevalence of mixed
Trichostrongylus (3.85%). Prevalence of different infection was found to be (23.33%) which Similar with
parasitic infestation in goat is shown in Table 1. More or Table 1. Prevalence of gastro-intestinal
less similar prevalence of gastrointestinal parasites have nematodiasis in goat
been reported earlier by Tariq et al., (2010), Lindqvist et Number

Sl. Name of Prevalence
of goat
al., (2001); Iqbal et al., (1993); Mcculloch et al., (1986) No. Parasites (%)
affected
and Khalid et al., (2004). Haemonchus
1 9 30.00a
contortus
In this study the dominant nematodes sp. was Strongyloides
2 5 16.67c
Haemonchus (30.00%) which was in agreement with papillosus
3 Trichuris ovis 3 10.00e
earlier reports by Iqbal et al., (1993); Yadav and Tandon Trichostrongylus
4 2 6.67f
(1989); Ahmad and Ansari (1987) and Cantreras et al., vitrinus
Oesophagostomum
(1976). The prevalence of Oesophagostomum (13.33%) 5 4 13.33d
columbianum
is in conformity with the report of Ijaz et al., (2008). 6 Mixed infection 7 23.33b
However, on the other hand, present finding of Total 30 100.00
LSD/Level of sig. - 2.15/**
Oesophagostomum is much lower than those reported by
* = Significant at 5% level of probability
Gupta et al., (1987). The prevalence of strongyloides sp. ** = Significant at 1% level of probability
(16.67%) is in conformity with the result observed by NS = Not significant

Akanda et al., 2012
the results recorded by Tariq et al., (2010). South Africa. J. South Africa Vet. Associ.,
The differences in the prevalence of various 61(3):112-116.
gastrointestinal parasitic infections in goats are thought Gupta RP, Yadav CL and Chaudhri SS. 1987.
to be due to sex variation, time of studies, risk factors Epidemiology of gastrointestinal nematodes of sheep and
and determinants, environmental condition (Mcculloch et goats in Haryana, India. Vet. Parasitol., 24(1-2):117-127.
al., 1986) and frequency of examination of the animal, Howell SB, Burke JM, Miller JE, Williamson LH,
management of helminthes status of a group of goats and Zajac AM and Kaplan RM. 2008. Prevalence of
anthelmintic resistance on sheep and goat farms in the
concurrent topography (Tariq et al., 2010). So, it is
southeastern United States. J Am Vet Med Assoc., 233
obviously a basic need to analyze the GIT in goats in this
(12):1913-9.
farm through cross-sectional studies to construct a
Husain SS, Amin MR and Islam ABMM. 1998. Goat
valuable epidemiological figure so that effective
production and its breeding strategy in Bangladesh. In
management and treatment can be introduced. proceedings of First National Workshop on Animal
breeding held in Bangladesh Agricultural University,
ACKNOWLEDGEMENT Mymensingh on November 26, 17-36.
The author is highly grateful to the personnel of Husnain HU and Usmani RH. 2006. Livestock of
Sylhet Govt. Goat Development Farm for their valuable Pakistan. 1st Ed. Livestock Foundation, Islamabad.
assist in conducting such survey. Ijaz M, Khan MS, Avais M, Ashraf K, Ali M and
Saima M. 2008. Infection rate and chemotherapy of
REFERENCES various helminths in goats in and around Lahore.
Ahmad M and Ansari JA. 1987. Prevalence of Pakistan Vet. J., 28(4):167-170.
gastro-intestinal nematodes of sheep and goats in
Ijaz M, Khan MS, Avais M, Ashraf K, Ali MM and
Aligarh. Indian Vet. Med. J., 11:165-170.
Khan MZU. 2009. Infection rate and chemotherapy of
Asanji MF and Wiliams MO. 1987. Variables affecting various helminthes in diarrhoeic sheep in and around
population dynamics of gastrointestinal helminth Lahore. J. Anim. and Plt. Sci., 19(1):13-16.
parasites of small farm ruminants in Sierra Leone. Bull.
Iqbal Z, Akhtar M, Khan MN and Riaz M. 1993.
Anim. Hlth. Prod. Afr., 35:308-313.
Prevalence and economic significance of haemonchosis
Blood DC, Henderson JA and Radostits OM. 1979. in sheep and goats slaughtered at Faisalabad abattoir.
Veterinary Medicine. 5th Ed., Bailliere Tindall, London, significance of haemonchosis in sheep and goats
UK. slaughtered at Faisalabad abattoir. Pakistan J. Agric. Sci.,
30:51-3.
Broomker J, Horak IG and Macivor KMDF. 1989.
Helminth parasites of grysbok common duikers and Kaplan RM, Burke JM, Terrill TH, Miller JE and
Angora and Boer goats in the valley bushveld in the Getz WR. 2004. Validation of the FAMACHA eye
Eastern Cape Province South Africa. Onderstepoort. J. color chart for detecting clinical anemia in sheep and
Vet. Res., 56(3):165-172. goats on farms in the southern united states. Vet.
Parasitol., 123:105-120.
Cantreras JA, Lopez W and Sanchez J. 1976.
Haemonchus infection in goats in Venezuela; a survey. Khalid SMA, Amin MR, Mostofa M, Hossain MJ and
Revista Veterinaria Venezolana 40(235):91-97. Azad MAK. 2004. Effects of Vermic against
Gastro-intestinal Nematodiasis in sheep. J. Biolog. Sci.,
Fivaz BH, Horak IG and Williams EJ. 1990. Helminth
4(6):720-724.
and arthropod parasites of Angora goats on irrigated
kikuyu grass pastures in the Eastern Capaae Province

Akanda et al., 2012
Lindqvist A, Ljungstrom BL, Nilsson O and Waller Waller PJ. 2006. Sustainable nematode parasite control
PJ. 2001. The dynamics, prevalence and impact of strategies for ruminant livestock by grazing management
nematode infections in organically raised sheep in and biological control. Anim. Feed Sci. Technol.,
Sweden. Acta Veterinaria Scandinavica 42:377-389. 126:277-289.
Mazid MA, Bhattacharjee J, Begum N and Rahman Waruiru RM, Ngotho JW, Mutune MN and
MH. 2006. Helminth parasites of the digestive system of Munyua WK. 2003. Comparative efficacy of
sheep in Mymensingh, Bangladesh. Bangl. J. Vet. Med., ivermectin, albendazole, levamisole and rafoxanide
4(2):117-122. against gastrointestinal nematode infections in goats.
Indian J. Ani. Sci., 73(2):147-150.
Mcculloch B, Dalbock RR and Kuehn HG. 1986. The
relation of climate and topography to gastrointestinal Yadav AK and Tandon V. 1989. Gastrointestinal
nematode worm egg counts of Angora goats in the nematodes infections of goats in a subtropical and humid
eastern cape South Africa. Onderstepoort J. Vet. Res., 53 zone of India. Vet. Parasitol., 33(2):135-142.
(3):167-178.
Zong Z, Wan-Zuomin, Shi, Jianthua, Siqin,
Perry BD, Randolph TF, McDermott JJ, Sones KR ZhaoRiGe, Zong-ZJ. 1997. Haemonchus contours,
and Thornton PK. 2002. Investing in a animal health Nematodirus and Oesophagostomum and their tissue
research to alleviate poverty. International Livestock stage Larvae in sheep. Chinese J. Vet. Med.,
Research Inst itute, Nairobi, Kenya, ISBN: 23(8):30-31.
92-9146-108-3, 148.
Raza MA, Murtaza S, Bachaya HA, Dastager G and

Hussain A. 2009. Point prevalence of haemonchosis in
sheep and goats slaughtered at multan abattoir. J. Ani. &
Plant Sci., 19(3):158-159.
Rehman KU, Javed K, Tunio MT and Kuthu ZH.

2009. Passive survalance of Gastro intestinal parasites in
buffaloes of mandi Bahauddin and Gujrat districts of the
Punjab. J. Animal & Plant Sci., 19(1):17-19.
Sahlu T, Dawson LJ, Gipson TA, Hart SP and

Merkel RC. 2009. ASAS Centennial Paper: Impact of
animal science research on United States goat production
and predictions for the future J. Anim. Sci., 87:400-418.
SAIC Newsletter. 1995. A publication of the SAARC

Agricultural Information Centre 5:2.
Silvestre A, Chartier C, Sauve C and Cabaret J. 2000. Submit your articles online at Ficuspublishers.com
Relationship between helminth species diversity, Advantages
intensity of infection and breeding management in dairy Easy online submission
goats. Vet. Parasitol. 94:91-105. Complete Peer review
Affordable Charges
Tariq KA, Chishti MZ, Ahmad F. 2010. Quick processing
Gastro-intestinal nematode infections in goats relative to Extensive indexing
season, host sex and age from the Kashmir valley. India J Open Access and Quick spreading
Helminthol., 84(1):93-7.

Curcumin and Cancer: Recent Developments

Authors: ABSTRACT:
Shyamasree Ghosh1,
Sanjima Pal1, Smita
Prusty1 and Girish KVS 2 . Cancer is a malignant disease with high mortality rates affecting millions.
Although chemotherapeutic agents are employed largely in cancer management, they
often result in toxicity and side effects and may lead to resistance. These draw backs
Institution: of the conventional chemotherapeutic agents have lead to the urging need for
1. School of Biological
development of safer, biocompatible, nontoxic compounds from natural sources and
Sciences.
their application in cancer management. Products from natural sources are being
2. School of Chemical exploited in cancer research worldwide due to its less toxicity. Of the several natural
Sciences. products tested, curcumin, well known for its chemopreventive, cytoprotective and
immune suppressive properties holds great promise for cancer research. Curcumin
National Institute of Science has been reported to affect different signaling pathways either in a direct or indirect
Education and Research, manner in a wide range of cancers. Despite the therapeutic effectiveness of curcumin,
Bhubaneswar, Institute its application is largely restricted due to its poor absorption, lipophilic nature and low
of Ph ysics Campus, bioavailability. Thus newer and effective formulations for curcumin in cancer
Sachivalaya Marg, treatment are being continuously exploited. In this review we highlighted (i) recent
PO: Sainik School, developments of application of natural products in cancer research (ii) role of
Bhubaneswar - 751 005, curcumin in different cancers (iii) curcumin formulations and their application in
India. cancer research. The future scope of this review lies in the effective employment of
curcumin and its formulations, in the eradication of cancer.
Corresponding author:
Shyamasree Ghosh.
Keywords:
Curcumin, cancer, anticancer therapy.

sree.s@niser.ac.in Shyamasree Ghosh, Sanjima Pal, SmitaPrusty and KVS Girish.
Curcumin and Cancer: Recent Developments.
Phone No:
0674-230-4049. Dates:
Received: 10 Mar 2012 /Accepted: 03 Apr 2012 /Published: 07 May 2012
Fax No :
0674-230-4070.
Ficus Publishers.
Web Address: This Open Access article is governed by the Creative Commons Attribution License (http://
http://jresearchbiology.com/ creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-
Documents/RA0211.pdf. cited.
251-272| JRB | 2012 | Vol 2 | No 3

Ghosh et al., 2012
INTRODUCTION 2004), through different signaling pathways portray it as

D e s pi t e on g oi n g pr ogr e s s of th e a promising chemotherapeutic molecule. The exact
chemotherapeutic agents in cancer management, it still molecular mechanism of curcumin in different cancer
remains to be a major killer. The main challenge in cells is being determined. However some of the target
cancer therapeutics revolves around specific killing of molecules that curcumin effect in different cancers
cancer cells within a milieu of the normal ones, include nuclear factor-B, activator protein-1, nitric
overcome resistance to chemotherapeutic drugs called as oxide synthase, receptor tyrosine kinases, cell cycle
Multi Drug Resistance (MDRs) and prevention of regulators like cyclins, matrix metalloproteinases, pro-
metastasis, targeting clinical relapse and augmentation of apoptotic markers and inhibits angiogenesis (Syng-Ai et
chemotoxicity. The conventional anticancer agents al., 2004; Aggarwal et al., 2007; Bierhaus et al., 1997;
employed in cancer management are mostly toxic that Brouet et al., 1995; Hahm et al., 2004; Korutla et al.,
lead to life threatening complications of neurotoxicity 1994; Singh et al., 1995). Curcumin is currently in
and cognitive changes (Dutta et al., 2011; Fardell et al., clinical trials for treatment of various cancers, including
2011). Therefore the need for safer, nontoxic products multiple myeloma, pancreatic cancer, and colon cancer
remains in targeting cancer. Different types of natural (Aggarwal et al., 2003).
products are being continuously exploited in cancer In this review we focus our attention to (a)
research (Ulbricht et al., 2010; Dai et al., 2010) due to mortality rates in different cancer (b) recent references to
the tested advantage of being less toxic than synthesized natural products applied in cancer research (c) how
chemical compounds. curcumin inhibits cancer through different pathways (d)
Curcumin, a major curcuminoid derivative, is a recent developments in curcumin formulations and
main component of turmeric obtained from the roots of applications in cancer research.
Curcuma longa and forms an important component of
the South Asian cushine. (Fig-1). It is chemically
formulated as diferuloylmethane (C21 H20 O6), a
polyphenol in its properties, curcumin has been reported
to affect the biological system immensely by its
antiinflammatory, antitumorigenic, antioxidant,
antiseptic, anti-toxic, cancer chemopreventive, chemo
sensitization, radio sensitization epigenetic change
inducer, and potentially chemotherapeutic properties
(Aggarwal et al., 2007, Basnet et al., 2011) and thus
Fig 1A: Plant Fig 1C: Rhizome
finds wide application in health research.
Several classes of bio-molecules are selectively
regulated by curcumin thereby affecting diverse
signaling pathways (Arora et al., 1973; Srimal et al.,
1973; Jobin et al., 1999; Sharma et al., 2005 and
Shanmugam et al., 2011) in the biological system. Fig 1B: Chemical Structure of curcumin
The hall mark property of targeted cell killing by Fig 1: Curcumin: 1A. Plant, 1B. Chemical Structure
of curcumin, 1C. Rhizome
curcumin (Shanmugam et al., 2011; Syng-Ai et al.,
Ghosh et al., 2012
Mortality rates in different types of cancer Hence the need for suitable molecules to fight the disease
Cancer remains to be a leading cause of death is urgent.
worldwide accounting for 7.6 million deaths (around Recent references of Natural products applied in
13% of all deaths) in 2008, as determined by statistics on cancer research
Cancer Incidence and Mortality Worldwide by A whole range of natural products obtained from
GLOBOCAN, Cancer Fact Sheet conducted by WHO bacterial, algal, fungal and higher plant sources
(http://www.globocon.iarc.fr/) (Fig-2). The statistics (Kinghorn et al., 2011) are being exploited for obtaining
summarized the mortality rate in cancer and was active molecules to combat cancer (Table-1).
represented by an Age-standardized rate (W) including Curcumin: Molecular Pathways and Inhibition of
the number of new cases or deaths per 100,000 persons Cancer
per year. In India the most frequent cancers reported Cancer is a disease with major manifestations
were cervix and uteri, breast, oral and lips, lungs and like uncontrolled cell proliferation and metastasis.
oesophagus while lung, breast, colorectal and prostrate Research in the field of cancer chemotherapy broadly
were the five most frequent occurring ones in the world. revolves around the control by the following mechanisms
Mortality Rate ASR (W)
Most Frequent Cancers

Fig 2: Cancer Statistics: World and India: The statistics is based on Mortality in most frequent cancers. The
data is based on Age-standardized rate (W): A rate is the number of new cases or deaths per 100, 000 persons
per year. An age-standardized rate is the rate that a population would have if it had a standard age structure.
(Globocon, WHO 2008, Cancer Facts Sheet).

Ghosh et al., 2012
(i) Inhibition of cell proliferation and metastasis (ii) retinoblastoma protein (Rb), and block clonogenicity of
Induced apoptosis (iii) Overcoming MDRs, (iv) Tumour tumor cells (Miller et al., 2008) thus enabling cancer
suppression. The effects of curcumin, in inducing the prevention.
above roles both by direct and indirect effect on In colon carcinoma, curcumin inhibits tumor
molecules have been extensively studied in different growth by increased apoptosis through the expression of
cancer and cell lines (Fig-3). Curcumin has been cyclooxygenase-2 (COX-2) and inhibition of
reported to play a dominant role in the prevention of proteasomal chymothypsin-like activity thereby leading
different types of cancer by promoting apoptosis or by to accumulation of ubiquitinated proteins and
inhibiting cell proliferation. proteasome target proteins IB-, p27, and p21/Bax
In pituitary cancer, curcumin has been reported leading to apoptosis (Milacic et al., 2008), activation of
to inhibit cell proliferation (Schaaf et al., 2009; Schaaf et Reactive Oxygen Species (ROS) (Lee YJ et al., 2011)
al., 2010) by decreased cyclin D3 expression thus and, suppression of mitochondrial NADP(+)-dependent
affecting cell cycle G1 to S transition and suppressed isocitrate dehydrogenase activity (Jung KH et al., 2011).
growth hormone (GH) levels like chymothypsin Curcumin also inhibits tumor cell proliferation by
prolactins and enhanced the growth-inhibitory effect of promoting cell cycle arrest in the G1 phase manifested
low concentrations of bromocriptine. It is also known to by decreased levels of PCNA, Cyclin D1, C-Myc, and
induce apoptosis by decreased phosphorylation of Bcl-2, Nuclear Factor kappa B, NF-kB-regulated
Fig 3: Curcumin and Cancer: Biomolecules and Pathways

Ghosh et al., 2012
Table 1: Natural Products and Cancer Research: Recent Developments
Major source of
Examples and References
Natural Product
Bacterial products (Wang C et al., 2011), Largazole, from the marine cyanobacterium
Bacteria Symploca species (Li S et al., 2011), Marine cyanobacterium Lyngbya majuscule (Tripathi
et al., 2011).
Algae Red sea weeds (Ahmed et al., 2011), Alga (Shalaby et al., 2011)
Endophytic fungal strain(Wang XN et al., 2011), Halophilic actinomycete
Fungus
Actinopolyspora erythraea YIM 90600(Zhao et al., 2011), Mushroom(Jiang et al., 2010).
Chinese herbs (Eichhorn et al., 2011), Thunder god vine or Tripterygium wilfordii Hook.
Herbs F. (Liu Z et al., 2011), Herbal flavenoids(Liu PX et al., 2011), lipid-soluble ginseng
extract (LSGE)(Kang et al, 2011), Phyllanthus urinaria (Huang ST et al., 2010).
Curcumin(Aggarwal et al., 2007; Basnet et al., 2011), Rhizome of Cnidium officinale(Bae
et al., 2011), Hypoestes forskaolii, Withania somnifera, Solanum glabratum, Adenium
obesum, Pistacia vera oleoresin, Caralluma quadrangula, Eulophia petersii,
Phragmanthera austroarabica, and Asparagus officinalis(Almehdar et al., 2011),
Terpenoids(Kuttan et al., 2011), Salvia officinalis L. (sage) essential oil(Sertel et al., 2011)
Xanthone V(1) and 2-acetylfuro-1,4-naphthoquinone(Kuete et al., 2011), Pomegranate
extract(Nair et al., 2011), Tea polyphenols(Singh M et al., 2011; Chen D et al., 2011),
green tea extract (Chen D et al., 2011; Cross et al., 2011; Vu et al., 2010; Lopez-Lazaro et
al., 2011; Liu X et al., 2011), catechins from green tea(Shimizu et al., 2011), methanolic
leaf extract of Indigofera cassioides (MEIC)(Kumar et al., 2011), phenolic compound
from the wood of Millettia leucantha(Rayanil et al., 2011), triterpenoid from the leaves of
Sinojackia sarcocarpa(Wang O et al., 2011), Phytochemicals green tea polyphenols
(epigallocatechin gallate)(Wahl et al., 2011), isoflavins from soy bean(Szliszka et al.,
2011; Jung et al., 2011), biflavonoid amentoflavone(Lee S et al., 2011), flesh of avocado
fruits(DAmbrosio et al., 2011), polyphenolic compounds isolated from the leaves of
Higher plants Leucenia leucocephala (Haggag et al., 2011), extracts of Xanthium strumarium
(Cocklebur)(Takeda et al., 2011), extracts from root bark of Juglans Regia L. (RBJR)
(Hasan et al., 2011), red beetroot (Beta vulgaris L.) extract(Kapadia et al., 2011), Moringa
oleifera leaf extract(Sreelatha et al., 2011), Solvent extracts from the aerial and root parts
and seed oil from E. sativa (rocket salad)(Khoobchandani et al., 2011), Toona sinensis
(leaf extracts)(Hseu et al., 2011), water extract from Mahonia bealei (Fort.) Carr. Leaves
(Hu et al., 2011), Palm tocotrienols(Selvaduray et al., 2010), Gugulipid (GL), extract from
medicinal plant Commiphora mukul (Xiao D et al., 2011), Methanol extracts of leaves of
Alnus sieboldiana(Ludwiczuk et al., 2011), ginger (Zingiber officinale Roscoe)
(Tuntiwechapikul et al., 2010), extract from black rice(Hui et al., 2010), Cacao
(Theobroma cacao L.)(Preza et al., 2010), Periwinkle (Catharanthus roseus)(Liscombe et
al., 2010), Panax stipuleanatus rhizomes(Liang et al., 2010), root extract of Polygala
senega(Paul et al., 2010), Black pepper (Piper nigrum) (Liu Y et al., 2010), organic
extracts of mulberry (Morus alba L.) leaves(Naowaratwattana et al., 2010), Phyllanthus
emblica L.(Ngamkitidechakul et al., 2010), dry olive leaf extract(Mijatovic et al., 2011),
Cedrus deodara lignins(Saxena et al., 2010).
Journal of Research in Biology (2012) 3:251-272 255

Ghosh et al., 2012
(Chen C et al., 2011). It is known to induce DNA damage angiogenesis via nuclear factor-kappa B pathway (Lin et
and cause S and G2/M arrest in the cell cycle (Lu JJ et al., al., 2007). Curcumin has been reported to induce
2012) . apoptosis in nasopharyngeal cancers by activation of
In breast cancer, curcumin is reported to prevent Reactive Oxygen Species (ROS), mitochondrial
tumor growth by causing cell cycle arrest by inhibiting depolarization and caspase 3 dependant pathways and
cyclin-dependent kinase (cdk) activity, suppressing prevents tumor cell growth and alters the phenotype of
cyclin D1 and cyclin E expression (Mukhopadhyay et migratory cells (Kuo et al., 2011; Wang et al., 2011a;
al., 2002), increasing levels of cdk inhibitors p21 and Wang et al., 2011b; Wong et al., 2010).
p27, inducing p53 transcriptional activity (Aggarwal et In liver cancer, curcumin has been reported to
al., 2007; Sen et al., 2011), inhibition of matrix cause cell death and promote mitochondria mediated
metalloproteinase-3 secretion (Boonrao et al., 2010) and apoptosis (Qian et al., 2011) and inhibits tumor cell
induce apoptosis by induction of Bax pathway growth (Cheng et al., 2010; Ning et al., 2009). Curcumin
(Choudhuri et al., 2002). It is also reported to cause has been reported to induce apoptosis in pancreatic
DNA damage and apoptosis in association with increased cancer (Sahu et al., 2009). to inhibit matrix
expression, phosphorylation, and cytoplasmic retention metalloproteinase protein-2, MMP-2 in human laryngeal
of the Breast cancer protein BRCA1 protein- a tumor squamous carcinoma cells (Mitra et al., 2006) promote
suppressor protein which is a critical mediator of DNA apoptosis in human lung adenocarcinoma cells (Zhang J
repair in response to double-strand breaks (Rowe et al., et al., 2010) by DNA damage (Saha et al., 2010), caspase
2009). pathways and ER Stress mechanisms (Wu et al., 2010).
In bladder cancer, curcumin has been reported to Curcumin has been reported to inhibit growth of uterine
prevent tumor cell growth and induces apoptosis (Saini (Tsuiji et al., 2011), ovarian cancer (Saydmohammed et
et al., 2011) via decreased expression of the proapoptotic al., 2010; Watson et al., 2010; Seo et al., 2010), brain
protein survivin and the angiogenic proteins vascular and nervous system cancer (Zanotto-Filho et al., 2011;
endothelial growth factor (VEGF) and VEGF receptor 1 Spiller et al., 2011). It is known to cause apoptosis and
(VEGFR1) (Chadalapaka et al., 2008). In Acute Myeloid prevent cell growth in Hodgekins lymphoma (Mackenzie
Leukemia (AML), curcumin promotes apoptosis and et al., 2008), lymphoma (Vishvakarma et al., 2011;
inhibit cell proliferation (Rao et al., 2011) and is reported Zhang et al., 2010; Li et al., 2009; Xiao et al., 2010;
to inhibit telomerase activity in human leukemia cell HL- Zhongguo et al., 2008) and esophageal cancers
60 (Mukherjee et al., 2007). In skin cancer, curcumin (OSullivan-Coyne et al., 2009; Tian et al., 2008). It is
inhibits tumor progression by inhibiting the mammalian reported to participate in mechanisms aiding in
target for rapamycin or mTOR pathway (Phillips et al., overcoming MDR in multiple myeloma (Xiao et al.,
2011). 2010). The overall effect of curcumin in different cancer
In gastric cancer, curcumin suppresses cancer has been summarized in Table-2.
cell proliferation and invasion via down-regulation of P Curcumin formulations and recent developments
21 activated kinase 1(PAK1) activity and cyclin D1 Although curcumin offers promise to cancer
expression (Cai et al., 2009) and overcome MDR (Tang research over the conventional toxic chemotherapeutic
et al., 2005) and inhibits proliferation by affecting the drugs, the main disadvantage of applicability of
cell cycle (Moragoda et al., 2001). In ovarian carcinoma curcumin in disease therapy is its lipophilic nature and
curcumin has been reported to inhibit tumor growth and poor aqueous solubility, minimum bioavailability and
Ghosh et al., 2012
Table 2: Major effects of Curcumin and its derivatives in different Cancers

Role of Target molecules/pathways
curcumin Cancer
Direct Indirect
p53-p300 cross-talk leading to cell death
Breast cancer(Sen et al., 2011)
ABC transporter pathway
TLR4 dimerization Proteasome pathway Multiple myeloma(Mujtaba et al., 2012)
Overcome IB kinase Gastric cancer cells
drug Down-regulating the activity of NF-B
Cyclooxygenase-2 (Tang et al., 2005; Yu LL et al., 2011)
resistance Protein kinase C Decrease of pro-caspase 3 pro-caspase 9,
Protein kinase A increase of PARP cleavage and the ratio of K562/A02(Lu JJ et al., 2012)
Phosphorylase kinase Bax/Bcl-xL
pp60c-src Demethylation of the Neurog1 gene and restored
tyrosine kinase the expression of this cancer-related Prostate cancer(Shu et al., 2011)
Ca2+-dependent CpG-methylation epigenome marker gene
Epigenetic protein kinase Down Regulation of histone deacetylases,
Xanthine oxidase histone acetyltransferases, DNA
Ca2+-ATPase of Cancer(Reuter et al., 2011)
methyltransferase I,
sarcoplasmic and miRNAs
reticulum Activation of Apoptosis pathways Prostate cancer(Yallapu et al., 2011b)
Inositol 1,4, Induce apoptosis by decreased phosphorylation
5-triphosphate Pituitary tumour(Schaaf et al, 2010)
of Retinoblastoma (Rb).
receptor Activation of ROS pathway, DNA damage Myelomonocytic leukemia
(Shrikanth et al,1994) mitochondria-mediated and ER stress-dependent (Huang et al., 2011)
pathways.
Down Regulation of caspase cascade Breast cancer cell line(Zong et al., 2011)
Down-regulation of Bcl-2 and procaspase-3 Adenocarcinoma cell line
and increased production of reactive oxygen (Ibrahim et al., 2008)
species (ROS) level
Apoptosis Increasing Bax expression, decreasing
the expression of Bcl-2 and Bcl-xL,
Small Cell Lung Cancer (Yang et al., 2011)
decreasing mitochondrial membrane
potential, increased ROS
ROS level Up regulation of Bax
Human nasopharyngeal carcinoma cells
and down regulation of Bcl-2,
(Kuo et al., 2011)
mitochondrial dysfunction
Chronic myeloid leukemia
Bcr-Abl suppression
(Acharya et al, 2011)
Down regulation of NF-B expression Oesophageal cancer (Tian et al., 2008)
Proapoptotic via cytotoxicity of Cum-np Rat C6 glioma cells (Shao et al., 2011)
FAS/caspase-8 (extrinsic) pathway and
ER stress proteins, growth arrest- and DNA Human non-small cell lung cancer cells
damage-inducible gene 153 (GADD153) (Wu et al., 2010)
and glucose-regulated protein 78 (GRP78)
Induction of apoptosis by cleavage of PARP, Gastric and colon cancer
caspase-3, and reduction in Bcl-XL levels (Moragoda et al., 2001)
Human mammary epithelial carcinoma
Inhibits Down-regulating the NF-kB transcription factor
MCF-7 cells (Zong et al., 2011)
invasion and
Down regulation of Matrix metalloproteinase-3 Human invasive breast carcinoma cells
metastasis
(MMP-3) (Yallapu et al, 2010b)
Induces a tumor-suppressive miRNA, miR-203 Bladder cancer (Rejinold et al., 2011c)
Reduces SCC-4 cell invasion, leads to Human tongue squamous cell carcinoma
the recruitment of alpha-tubulin. (Chen JW et al,, 2011)
Inhibits tumor growth by inhibiting angiogenesis Colon cancer (Gou et al., 2011)
Inhibition of STAT3 signaling pathway
Tumor Human rhabdomyosarcoma cells
via LLL12 and FLLL32,
suppression (Wei et al., 2011)
down regulation of cyclin D1, Bcl-xL
Inhibits tumor cell proliferation Jurkat cell lines (Yadav et al., 2010)
Inhibition of Activation of nuclear factor-kappaB Pancreatic cancer (Bisht et al., 2010)
Disturbed mitotic spindle structure,
MCF-7 cells (Banerjee et al., 2010)
activated mitotic check points
Drug resistant AML cell lines
G1/S arrest
(Rao et al., 2011)
Inhibition of NF-kB signaling Medulloblastoma (Spiller et al., 2011)
targeted delivery to the transformed cell.

Ghosh et al., 2012
Different formulations and delivery devices are being Das et al., 2010), aerosols (Selvam et al., 2011), and
tested to enable curcumin as an effective agent in cancer nanodisks (Singh AT et al., 2011; Tadmor et al., 2011;
management. Biocompatible urithin polymers (PU) from Ghosh et al., 2011) are being tested as delivery
polylactic acids and hexamethylene diamide (Selvaraj et formulations of curcumin in different cancer.
al., 2011), liposomes coated with N-trimethyl chitosan Few reports on targeted delivery (Thamake et al.,
chloride (TMC) (Chen H et al., 2011) have been reported 2011) of curcumin exists. Tissue specific targeting are
to function as efficient formulations. Cationic and being tested like curcumin loaded Eudragit S100
anionic curcumin conjugates by anchoring curcumin coated calcium pectinate microsphere in colon cancer
(Cur) onto poly (vinylpyrroidone) (PVP-Cur) and onto (Zhang L et al., 2011). Reports on the use of Gelatin
hyaluronic acid (HA-Cur) (Manju et al., 2011b) are also microspheres (C-GMS) in lung cancer (Cao et al., 2011)
being exploited. Delivery through formulations like and Gelucire44/14 in eyes (Liu R et al., 2011) have
niosomes (Rungphanichkul et al., 2011), amphiphilic shown promising results. The application of different
polymers like Lauroyl sulphated chitosan (LSCS) formulations of curcumin in different cancer is
(Shelma et al., 2011) and carboxy methyl derivatives summarized in Table-3.
(Anitha et al., 2011), fibrinogen nanoparticles (CRC-
FNPs) (Rejinold et al., 2011a), nanospheres (Mukerjee DISCUSSIONS
et al., 2009), 2-Hydroxypropyl--cyclodextrin/curcumin- Curcumin has shown strong reports in its anti-
liposomal nanoparticles (Dhule et al., 2011) have been cancerous activities in different malignancies including
reported to be effective. Nanoparticles (Li R et al., 2011; brain, skin, lung, prostate, breast, ovarian, liver,
Yallapu et al., 2010a; Yallapu et al., 2010b) reported for nasopharyngeal, gastrointestinal, pancreatic and
their biocompatible, non toxic, biodegradable nature and colorectal cancers. Synergistic effects of curcumin with
thermoresponsive properties (Rejinold et al., 2011c) have agents like decetaxel has also been reported in lung
shown positive results in curcumin delivery. Conjugated cancer (Yin et al., 2011). However, because of its both
nanoparticle copolymers with Chitosan-g-poly (N- pro and anti-oxidant effect, it has been reported to
isopropylacrylamide) nanoparticles (Rejinold et al., behave like a double edged sword. (Kawanishi et al.,
2011b), amphilic methoxy polyethylene glycol-poly 2005) and its safety as a chemopreventive agent remains
(caprolactone) (mPEG-PCL) are being employed (Shao yet to be exploited. The future scope of this review
et al., 2011). Magnetic particles are being tested for remains in potential applications of targeted cancer cell
controlled drug delivery (Koppolu et al., 2010; Chin et killing by a natural product like curcumin.
al., 2010; de-Souza et al., 2011).
Strategies to increase the solubility of analogues ACKNOWLEDGEMENT
of curcumin (Zhang et al., 2011), lipid-based The study was conducted in the existing facility
formulations (Thangapazham et al., 2008; Yu et al., of the School of Biological Sciences, SBS, National
2011; Setthacheewakul et al., 2011; Xie et al., 2011), Institute of Science Education and Research (NISER),
nanosuspensions (Zhang H et al, 2011), liposomes (Chen Bhubaneswar, DAE, Govt. of India.
H et al., 2011; Dhule et al., 2011; Li et al., 2005; Mach
et al, 2009; Pandelidou et al., 2011; Agashe et al., 2011),
microemulsions (Liu CH et al., 2011), polymer
encapsulations (Mohanty et al., 2010; Sahu et al., 2011;
Ghosh et al., 2012
Table 3: Curcumin and Formulations : Cancer Research
Cancer Delivery Systems
Lung adenocarcinoma H441 cells and nude Cyclodextrin entrapped curcuminoid derivative(Agashe et al., 2011)
rats bearing xenograft H441 tumors
Micro emulsions for transdermal delivery(Liu et al., 2011)

Skin cancer
Non small cell lung cancer
Intravenous synergistic with Docetaxel(Yin et al., 2011)
Matrix of Urethane polymers (PU) prepared from low-molecular
Lung cancer cells weight polylactic acid (PLA) and hexamethylene diisocyanate
(HMDI) curcumin-containing PU membranes(Selvaraj et al., 2011)
Glioma cells and Caco-2 cells
Magnetic nanoparticles (MNPs) (Manju et al., 2011a)
L929 (mouse fibroblast), PC3 (prostate)
and MCF7 (breast) cancer cell lines Fibrinogen nanoparticles (Rejinold et al., 2011b)
Curcuminoids-loaded solid lipid nanoparticles (curcuminoids-SLNs) and

Tumor mice model curcumin-loaded solid lipid nanoparticles (curcumin-SLNs)(Li et al., 2005)
Osteosarcoma Liposomal nanoparticles(Dhule et al., 2011)
Cancer cells N,O-Carboxymethyl Chitosan Nanoparticles(Anitha et al., 2011)

Bcr-Abl + leukemia cells Nanoparticles(Acharya et al, 2011)
PC3, L929 cells Biocompatible thermoresponsive polymeric chitosan-g-poly
(N-vinylcaprolactam) nanoparticles (TRC-NPs)(Rejinold et al., 2011c)
Colorectal cell lines Lyophilised egg PC liposomes (Pandelidou et al., 2011)
Methoxy polyethylene glycol-poly (caprolactone) nanoparticles

C6 Glioma cells
(Shao et al, 2011)
3T3-L1 preadipocytes and adipocytes Conjugated with Polyethylene glycol (Kim et al., 2011)
Surface functionalized polymeric PLGA nanoparticles by non-covalent
Breast cells insertion of a homo-bifunctional chemical crosslinker,
bis(sulfosuccinimidyl) suberate (BS3) for targeted cancer therapy
(Thamake et al., 2011)
L929 cells Hollow microcapsules (Manju et al., 2011 b)
Cisplatin resistant A2780CP ovarian and

Encapsulated PGLA formulation (Yallapu et al., 2010a)
metastatic MDA-MB-231 breast cancer
cells
Poly(-cyclodextrin)/curcumin self-assembly (Yallapu et al., 2010b),
Prostrate cancer Nanoparticles (Thangapazham et al, 2008), cellulose nanoparticles
(Yallapu et al.,, 2011a), PGLA nanospheres (Mukerjee et al., 2009)
A biodegradable and biocompatible polymer, poly (d,l-lactide-co-glycolide),
Sub cutaneous injection in mice
was used to fabricate curcumin microparticles (Shahani et al., 2010)
Encapsulated methoxy poly (ethylene glycol)
Pancreatic cell lines MIA PaCa-2 (MePEG)/poly-epsilon-caprolactone (PCL)
and PANC-1 diblock copolymeric micelle (Mohanty et al., 2010)
Micelles of Pluronic encapsulated curcumin (Sahu et al., 2011),
Hela cells Alginate-chitosan-pluronic composite nanoparticles (Das et al., 2010)
Pancreatic cancer Liposome mediated (Li et al., 2005)

Ghosh et al., 2012
REFERENCES Arora RB, Kapoor V, Basu N, and Jain AP. 1973.

Acharya S, Sahoo SK. 2011. Sustained targeting of Bcr- Anti-inflammatory studies on Curcuma longa (turmeric).
Abl + leukemia cells by synergistic action of dual drug Indian J Med Res., 59:1289-1295.
loaded nanoparticles and its implication for leukemia
Bae KE, Choi YW, Kim ST, Kim YK. 2011.
therapy. Biomaterials., 32:5643-5662.
Components of Rhizome Extract of Cnidium officinale
Agashe H, Sahoo K, Lagisetty P, Awasthi V. 2011. Makino and Their In vitro Biological Effects.
Cyclodextrin-mediated entrapment of curcuminoid 4-[3,5 Molecules., 16:8833-8847.
-bis(2-chlorobenzylidene-4-oxo-piperidine-1-yl)-4-oxo-2
Banerjee M, Singh P, and Panda D. 2010. Curcumin
-butenoic acid] or CLEFMA in liposomes for treatment
suppresses the dynamic instability of microtubules,
of xenograft lung tumor in rats. Colloids Surf B
activates the mitotic checkpoint and induces apoptosis in
Biointerfaces., 84:329-337.
MCF-7 cells. FEBS J., 277(16):3437-3448.
Aggarwal BB, Banerjee S, Bharadwaj U, Sung B,
Basnet P, Skalko-Basnet N. 2011. Curcumin: An anti-
Shishodia S, Sethi G. 2007. Curcumin induces the
inflammatory molecule from a curry spice on the path to
degradation of cyclin E expression through ubiquitin-
cancer treatment. Molecules., 16:4567-4598.
dependent pathway and up-regulates cyclin-dependent
kinase inhibitors p21 and p27 in multiple human tumor Bierhaus A, Zhang Y, Quehenberger P, Luther T,
cell lines. Biochem Pharmacol., 73:1024-1032. Haase M, Mller M, Mackman N, Ziegler R,
Nawroth PP. 1997. The dietary pigment curcumin
Aggarwal BB, Kumar A, Bharti AC. 2003. Anticancer
reduces endothelial tissue factor gene expression by
potential of curcumin: preclinical and clinical studies.
inhibiting binding of AP-1 to the DNA and activation of
Anticancer Res., 23:363-398.
NF-kappa B. Thromb Haemost., 77:772-782.
Aggarwal Bharat B, Surh Young-Joon, Shishodia
Bisht S, Mizuma M, Feldmann G, Ottenhof NA, Hong
Shishir. 2007. The molecular targets and therapeutic
SM, Pramanik D, Chenna V, Karikari C, Sharma R,
uses of curcumin in health and disease. Springer Science
Goggins MG, Rudek MA, Ravi R, Maitra A, Maitra
pub., 1-47.
A. 2010. Systemic administration of polymeric
Ahmed HH, Hegazi MM, Abd-Alla HI, Eskander EF, nanoparticle-encapsulated curcumin (NanoCurc) blocks
Ellithey MS. Z. 2011. Antitumour and antioxidant tumor growth and metastases in preclinical models of
activity of some Red Sea seaweeds in Ehrlich ascites pancreatic cancer. Mol Cancer Ther., 9:2255-2264.
carcinoma in vivo. Z Naturforsch C., 66:367-376.
Boonrao M, Yodkeeree S, Ampasavate C,
Almehdar H, Abdallah HM, Osman AM, Abdel- Anuchapreeda S, Limtrakul P. 2010. The inhibitory
Sattar EA. 2011. In vitro cytotoxic screening of selected effect of turmeric curcuminoids on matrix
Saudi medicinal plants. J Nat Med., Sep 28. metalloproteinase-3 secretion in human invasive breast
carcinoma cells. Arch Pharm Res., 33:989-998.
Anitha A, Maya S, Deepa N, Chennazhi KP, Nair SV,
Jayakumar R. 2011. Curcumin-Loaded N,O- Brouet I, Ohshima H. 1995. Curcumin, an anti-tumour
Carboxymethyl Chitosan Nanoparticles for Cancer Drug promoter and anti-inflammatory agent, inhibits induction
Delivery, J Biomater Sci Polym Ed. Jun 28. of nitric oxide synthase in activated macrophages,

Ghosh et al., 2012
Biochem Biophys Res Commun., 206:533-540. Chin SF, Iyer KS, Saunders M, St Pierre TG,
Buckley C, Paskevicius M, Raston CL. 2010.
Cai XZ, Wang J, Li XD, Wang GL, Liu FN, Cheng
Encapsulation and sustained release of curcumin using
MS, and Li F. 2009. Curcumin suppresses proliferation
superparamagnetic silica reservoirs. Chemistry., 15:5661
and invasion in human gastric cancer cells by
-5665.
downregulation of PAK1 activity and cyclin D1
expression. Cancer Biol Ther., 8:1360-1368. Choudhuri T, Pal S, Agwarwal ML, Das T and Sa G.
2002. Curcumin induces apoptosis in human breast
Cao F, Ding B, Sun M, Guo C, Zhang L, Zhai G.
cancer cells through p53-dependent Bax induction. FEBS
2011. Lung-targeted delivery system of curcumin loaded
Lett., 512:334-340.
gelatin microspheres. Drug Deliv 18:545-554.
Cross SE, Jin YS, Lu QY, Rao J, Gimzewski JK.
Chadalapaka G, Jutooru I, Chintharlapalli S,
2011. Green tea extract selectively targets
Papineni S, Smith R , Li X, Safe S. 2008. Curcumin
nanomechanics of live metastatic cancer cells.
decreases specificity protein expression in bladder cancer
Nanotechnology., 22:215101.
cells. Cancer Res 68:5345-5354.
Dai J, Mumper RJ. 2010. Plant phenolics: extraction,
Chen C, Liu Y, Chen Y, Xu J. 2011. C086, a novel
analysis and their antioxidant and anticancer properties.
analog of curcumin, induces growth inhibition and down
Molecules., 15:7313-7352.
-regulation of NFB in colon cancer cells and xenograft
tumors. Cancer Biol Ther.,12:797-807. D'Ambrosio SM, Han C, Pan L, Kinghorn AD, Ding
H. 2011. Aliphatic acetogenin constituents of avocado
Chen D, Wan SB, Yang H, Yuan J, Chan TH, Dou
fruits inhibit human oral cancer cell proliferation by
QP. 2011. EGCG, green tea polyphenols and their
targeting the EGFR/RAS/RAF/MEK/ERK1/2 pathway.
synthetic analogs and prodrugs for human cancer
Biochem Biophys Res Commun., 409:465-469.
prevention and treatment. Adv Clin Chem., 53:155-177.
Das RK, Kasoju N, Bora U. 2011. Encapsulation of
Chen H, Wu J, Sun M, Guo C, Yu A, Cao F, Zhao L,
curcumin in alginate-chitosan-pluronic composite
Tan Q, Zhai G. 2011. N-trimethyl chitosan chloride-
nanoparticles for delivery to cancer cells.
coated liposomes for the oral delivery of curcumin. J
Nanomedicine., 6:153-160.
Liposome Res., Oct 18.
De Souza FF, dos Santos MC, dos Passos DC, Lima
Chen JW, Tang YL, Liu H, Zhu ZY, L D, Geng N,
EC, Guillo LA. 2011. Curcumin associated magnetite
Chen Y. 2011. [Anti-proliferative and anti-metastatic
nanoparticles inhibit in vitro melanoma cell growth. J
effects of curcumin on oral cancer cells]. [Article in
Nanosci Nanotechnol., 11:7603-7610.
Chinese] Hua Xi Kou Qiang Yi Xue Za Zhi., 29:83-86.
Dhule SS, Penfornis P, Frazier T, Walker R, Feldman
Cheng CY, Lin YH, Su CC. 2010. Curcumin inhibits
J, Tan G, He J, Alb A, John V, Pochampally R. 2011.
the proliferation of human hepatocellular carcinoma J5
Curcumin-loaded -cyclodextrin liposomal nanoparticles
cells by inducing endoplasmic reticulum stress and
as delivery vehicles for osteosarcoma. Nanomedicine.,
mitochondrial dysfunction. Int J Mol Med., 26:673-678.
Aug 10.

Ghosh et al., 2012
Dutta V. 2011. Chemotherapy, neurotoxicity, and endothelial growth factor (VEGF)-induced angiogenesis
cognitive changes in breast cancer J Cancer Res Ther., in vascular endothelial cells. J Ethnopharmacol., 134:111
7:264-269. -121. http://www.globocan.iarc.fr/.
Eichhorn T, Efferth T. 2011. P-glycoprotein and its Hu W, Yu L, and Wang MH. 2011. Antioxidant and
inhibition in tumors by phytochemicals derived from antiproliferative properties of water extract from
Chinese herbal medicine, J Ethnopharmacol., Mahonia bealei (Fort.) Carr. Leaves. Food Chem
Toxicol., 49:799-806.
Fardell JE, Vardy J, Johnston IN, Winocur G. 2011.
Chemotherapy and cognitive impairment: treatment Huang AC, Chang CL, Yu CS, Chen PY, Yang JS, Ji
options , Clin Pharmacol Ther., 90:366-376. BC, Lin TP, Chiu CF, Yeh SP, Huang YP, Lien JC,
Chung JG. 2011. Induction of apoptosis by curcumin in
Ghosh M, Singh AT, Xu W, Sulchek T, Gordon LI,
murine myelomonocytic leukemia WEHI-3 cells is
Ryan RO. 2011. Curcumin nanodisks: formulation and
mediated via endoplasmic reticulum stress and
characterization, Nanomedicine., 7:162-167.
mitochondria-dependent pathways, Environ Toxicol.,
Gou M, Men K, Shi H, Xiang M, Zhang J, Song J, Jul 26.
Long J, Wan Y, Luo F, Zhao X, Qian Z. 2011.
Huang ST, Pang JH, and Yang RC. 2010. Anti-cancer
Curcumin-loaded biodegradable polymeric micelles for
effects of Phyllanthus urinaria and relevant mechanisms.
colon cancer therapy in vitro and in vivo. Nanoscale.,
Chang Gung Med J., 3:477-487.
3:1558-1567.
Hui C, Bin Y, Xiaoping Y, Long Y, Chunye C,
Haggag EG, Kamal AM, Abdelhady MI, El-Sayed
Mantian M, Wenhua L. 2010. Anticancer activities of
MM, El-Wakil EA, Abd-El-Hamed SS. 2011.
an anthocyanin-rich extract from black rice against breast
Antioxidant and cytotoxic activity of polyphenolic
cancer cells in vitro and in vivo. Nutr Cancer., 62:1128-
compounds isolated from the leaves of Leucenia
1136.
leucocephala. Pharm Biol., 49:1103-1113.
Ibrahim A, El-Meligy A, Lungu G, Fetaih H,
Hahm ER, Gho YS, Park S, Park C, Kim KW, Yang
Dessouki A, Stoica G, Barhoumi R. 2008. Curcumin
CH. 2004. Synthetic curcumin analogs inhibit activator
induces apoptosis in a murine mammary gland
protein-1 transcription and tumor-induced angiogenesis.
adenocarcinoma cell line through the mitochondrial
Biochem Biophys Res Commun., 321:337-344.
pathway. Eur J Pharmacol., 668:127-132.
Hasan TN, B LG, Shafi G, Al-Hazzani AA, Alshatwi
Jiang J, Sliva D. 2010. Novel medicinal mushroom
AA. 2011. Anti-proliferative effects of organic extracts
blend suppresses growth and invasiveness of human
from root bark of Juglans Regia L. (RBJR) on MDA-MB
breast cancer cells. Int J Oncol., 37:1529-1536.
-231 human breast cancer cells: role of Bcl-2/Bax,
caspases and Tp53. Asian Pac J Cancer Prev., 12:525- Jobin C, Bradham CA, Russo MP, Juma B, Narula
530. AS, Brenner DA, Sartor RB. 1999. Curcumin blocks
cytokine-mediated NF-kappa B activation and
Hseu YC, Chen SC, Lin WH, Hung DZ, Lin MK,
proinflammatory gene expression by inhibiting inhibitory
Kuo YH, Wang MT, Cho HJ, Wang L, Yang HL.
factor I-kappa B kinase activity. J Immunol., 163:3474-
2011. Toona sinensis (leaf extracts) inhibit vascular
Ghosh et al., 2012
3483. Koppolu B, Rahimi M, Nattama S, Wadajkar A,

Nguyen KT. 2010. Development of multiple-layer
Jung KH, Park JW. 2011. Suppression of
polymeric particles for targeted and controlled drug
mitochondrial NADP (+)-dependent isocitrate
delivery. Nanomedicine., 6:355-361.
dehydrogenase activity enhances curcumin-induced
apoptosis in HCT116 cells. Free Radic Res., 45:431-438. Korutla L, Kumar R. 1994. Inhibitory effect of
curcumin on epidermal growth factor receptor kinase
Jung MY, Choi NJ, Oh CH, Shin HK, and Yoon SH.
activity in A431 cells. Biochim Biophys Acta., 1224:597
2011. Selectively hydrogenated soybean oil exerts strong
-600.
anti-prostate cancer activities. Lipids, 46:287-295.
Kuete V, Wabo HK, Eyong KO, Feussi MT, Wiench
Kang MR, Kim HM, Kang JS, Lee K, Lee SD, Hyun
B, Krusche B, Tane P, Folefoc GN, Efferth T. 2011.
DH, In MJ, Park SK, Kim DC. 2011. Lipid-soluble
Anticancer activities of six selected natural compounds
ginseng extract induces apoptosis and G0/G1 cell cycle
of some Cameroonian medicinal plants, PLoS One.,
arrest in NCI-H460 human lung cancer cells. Plant Foods
6:e21762.
Hum Nutr., 66:101-106.
Kumar RS, Rajkapoor B, Perumal P. 2011. In vitro
Kapadia GJ, Azuine MA, Rao GS, Arai T, Iida A,
and in vivo anticancer activity of Indigofera cassioides
Tokuda H. 2011. Cytotoxic effect of the red beetroot
Rottl. Ex. DC. Asian Pac J Trop Med., 4:379-385.
(Beta vulgaris L.) extract compared to doxorubicin
(Adriamycin) in the human prostate (PC-3) and breast Kuo CL, Wu SY, Ip SW, Wu PP, Yu CS, Yang JS,
(MCF-7) cancer cell lines. Anticancer Agents Med Chen PY, Wu SH, Chung JG. 2011. Apoptotic death in
Chem., 11:280-284. curcumin-treated NPC-TW 076 human nasopharyngeal
carcinoma cells is mediated through the ROS,
Kawanishi S, Oikawa S, Murata M. 2005. Evaluation
mitochondrial depolarization and caspase-3-dependent
for safety of antioxidant chemopreventive agents.
signaling responses. Int J Oncol., 39:319-328.
Antioxid Redox Signal.7:1728-39.
Kuttan G, Pratheesh kumar P, Manu KA, Kuttan R.
Khoobchandani M, Ganesh N, Gabbanini S,
2011. Inhibition of tumor progression by naturally
Valgimigli L, Srivastava MM. 2011. Phytochemical
occurring terpenoids, Pharm Biol., 49:995-1007.
potential of Eruca sativa for inhibition of melanoma
tumor growth. Fitoterapia .,82:647-653. Lee S, Kim H, Kang JW, Kim JH, Lee DH, Kim MS,
Yang Y, Woo ER, Kim YM, Hong J,Yoon DY. 2011.
Kim CY, Bordenave N, Ferruzzi MG, Safavy A, Kim
The biflavonoid amentoflavone induces apoptosis via
KH. 2011. Modification of curcumin with polyethylene
suppressing E7 expression cell cycle arrest at sub-G
glycol enhances the delivery of curcumin in
phase, and mitochondria-emanated intrinsic pathway in
preadipocytes and its antiadipogenic property. J Agric
human cervical cancer cells. J Med Food., 14:808-816.
Food Chem., 59:1012-1019.
Lee YJ, Kim NY, Suh YA, Lee C. 2011. Involvement
Kinghorn AD, Pan L, Fletcher JN, Chai H. 2011. The
of ROS in Curcumin-induced Autophagic Cell Death.
relevance of higher plants in lead compound discovery
Korean J Physiol Pharmacol., 15:1-7.
programs V J Nat Prod .,74:1539-1555.

Ghosh et al., 2012
Lee-Sherick AB, Linger RM, Gore L, Keating AK, Liscombe DK, Usera AR, and OConnor SE. 2010.
Graham DK. 2010. Targeting paediatric acute Homolog of tocopherol C methyltransferases catalyzes N
lymphoblastic leukaemia: novel therapies currently in methylation in anticancer alkaloid biosynthesis. Proc
development. Br J Haematol., 151:295-311. Natl Acad Sci U S A., 107:18793-18798.
Li L, Braiteh FS, Kurzrock R. 2005. Liposome- Liu CH, Chang FY. 2011. Development and
encapsulated curcumin: in vitro and in vivo effects on characterization of eucalyptol microemulsions for topic
proliferation, apoptosis, signaling, and angiogenesis. delivery of curcumin, Chem Pharm Bull (Tokyo).,
Cancer., 104:1322-1331. 59:172-178.
Li R, Qiao X, Li Q, He R, Ye M, Xiang C, Lin X, Guo Liu PX, Gao J, Chen YJ, Long W, Shen X, Tang WS.
D. 2011. Metabolic and pharmacokinetic studies of 2011. Anticancer activity of total flavonoids isolated
curcumin, demeth oxycur cumin and from Xianhe Yanling Recipe. Chin J Integr Med., 17:459
bisdemethoxycurcumin in mice tumor after intragastric -463.
administration of nanoparticle formulations by liquid
Liu R, Liu Z, Zhang C, Zhang B. 2011. Gelucire44/14
chromatography coupled with tandem mass
as a novel absorption enhancer for drugs with different
spectrometry. J Chromatogr B Analyt Technol Biomed
hydrophilicities: in vitro and in vivo improvement on
Life Sci., 879:2751-2758.
transcorneal permeation. J Pharm Sci., 100:3186-3195.
Li S, Yao H, Xu J, Jiang S. 2011. Synthetic routes and
Liu X, Zhang DY, Zhang W, Zhao X, Yuan C, Ye F.
biological evaluation of largazole and its analogues as
2011. The effect of green tea extract and EGCG on the
potent histone deacetylase inhibitors, Molecules.,
signaling network in squamous cell carcinoma. Nutr
16:4681-4694.
Cancer., 63:466-475.
Li ZX, Ouyang KQ, Jiang X, Wang D, Hu Y. 2009.
Liu Y, Yadev VR, Aggarwal BB, Nair MG. 2010.
Curcumin induces apoptosis and inhibits growth of
Inhibitory effects of black pepper (Piper nigrum) extracts
human Burkitt's lymphoma in xenograft mouse model.
and compounds on human tumor cell proliferation,
Mol Cells., 27:283-289.
cyclooxygenase enzymes, lipid peroxidation and nuclear
Liang C, Ding Y, Nguyen HT, Kim JA, Boo HJ, Kang transcription factor-kappa-B. Nat Prod Commun., 5:1253
HK, Nguyen MC, Kim YH. 2010. Oleanane-type -1257.
triterpenoids from Panax stipuleanatus and their
Liu Z, Ma L, and Zhou GB. 2011. The main anticancer
anticancer activities, Bioorg Med Chem Lett., 20:7110-
bullets of the Chinese medicinal herb, thunder god vine,
7115.
Molecules., 16:5283-5297.
Lin YG, Kunnumakkara AB, Nair A, Merritt WM,
Lpez-Lzaro M, Caldern-Montao JM,
Han LY, Armaiz-Pena GN, Kamat AA, Spannuth
Burgos-Morn E, Austin CA. 2011. Green tea
WA, Gershenson DM, Lutgendorf SK, Aggarwal BB,
constituents (-) epigallocatechin-3-gallate (EGCG) and
Sood AK. 2007. Curcumin inhibits tumor growth and
gallic acid induce topoisomerase I- and topoisomerase II-
angiogenesis in ovarian carcinoma by targeting the
DNA complexes in cells mediated by pyrogallol-induced
nuclear factor-kappaB pathway. Clin Cancer Res.,
hydrogen peroxide, Mutagenesis., 26:489-498.
13:3423-3430.
Ghosh et al., 2012
Lu JJ, Cai YJ, Ding J. 2012. The short-time treatment the proteasome activity in human colon cancer cells in
with curcumin sufficiently decreases cell viability, vitro and in vivo. Cancer Res., 68:7283-7292.
induces apoptosis and copper enhances these effects in
Miller M, Chen S, Woodliff J, Kansra S. 2008.
multidrug-resistant K562/A02 cells. Mol Cell Biochem.,
Curcumin (diferuloylmethane) inhibits cell proliferation,
360:253-260.
induces apoptosis, and decreases hormone levels and
Ludwiczuk A, Saha A, Kuzuhara T, Asakawa Y. secretion in pituitary tumor cells. Endocrinology.,
2011. Bioactivity guided isolation of anticancer 149:4158-4167.
constituents from leaves of Alnus sieboldiana
Mitra A, Chakrabarti J, Banerji A, Chatterjee A and
(Betulaceae). Phytomedicine., 18:491-498.
Das BR. 2006. Curcumin, a potential inhibitor of MMP-
Mach CM, Mathew L, Mosley SA, Kurzrock R, 2 in human laryngeal squamous carcinoma cells HEp2. J
Smith JA. 2009. Determination of minimum effective Environ Pathol Toxicol Oncol., 25:679-690.
dose and optimal dosing schedule for liposomal
Mohanty C, Acharya S, Mohanty AK, Dilnawaz F,
curcumin in a xenograft human pancreatic cancer model.
Sahoo SK. 2010. Curcumin-encapsulated MePEG/PCL
Anticancer Res., 29:1895-1899.
diblock copolymeric micelles: a novel controlled
Mackenzie GG, Queisser N, Wolfson ML, Fraga CG, delivery vehicle for cancer therapy. Nanomedicine
Adamo AM, Oteiza PI. 2008. Curcumin induces cell- (Lond)., 5:433-449.
arrest and apoptosis in association with the inhibition of
Moragoda L, Jaszewski R, Majumdar AP. 2001.
constitutively active NF-kappaB and STAT3 pathways in
Curcumin induced modulation of cell cycle and
Hodgkin's lymphoma cells, Int J Cancer 123:56-65.
apoptosis in gastric and colon cancer cells. Anticancer
Manju S, Sreenivasan K. 2011a. Enhanced Drug Res., 21:873-878.
Loading on Magnetic Nanoparticles by Layer-by-Layer
Mujtaba T, Kanwar J, Wan SB, Chan TH, Dou QP.
Assembly Using Drug Conjugates: Blood Compatibility
2012. Sensitizing human multiple myeloma cells to the
Evaluation and Targeted Drug Delivery in Cancer Cells.
proteasome inhibitor bortezomib by novel curcumin
Langmuir., 27:14489-14496.
analogs, Int J Mol Med., 29:102-106.
Manju S, Sreenivasan K. 2011b. Hollow microcapsules
Mukerjee A, Vishwanatha JK. 2009. Formulation,
built by layer by layer assembly for the encapsulation
characterization and evaluation of curcumin-loaded
and sustained release of curcumin, Colloids Surf B
PLGA nanospheres for cancer therapy. Anticancer Res.,
Biointerfaces., 82:588-593.
29:3867-3875.
Mijatovic SA, Timotijevic GS, Miljkovic DM,
Mukherjee Nee Chakraborty S, Ghosh U,
Radovic JM, Maksimovic-Ivanic DD, Dekanski DP,
Bhattacharyya NP, Bhattacharya RK, Dey S,Roy M.
Stosic-Grujicic SD. 2011. Multiple antimelanoma
2007. Curcumin-induced apoptosis in human leukemia
potential of dry olive leaf extract. Int J Cancer., 128:1955
cell HL-60 is associated with inhibition of telomerase
-1965.
activity. Mol Cell Biochem., 297:31-39.
Milacic V, Banerjee S, Landis-Piwowar KR, Sarkar
Mukhopadhyay A, Banerjee S, Stafford LJ, Xia C,
FH, Majumdar AP, Dou QP. 2008. Curcumin inhibits
Liu M, Aggarwal BB. 2002. Curcumin-induced

Ghosh et al., 2012
suppression of cell proliferation correlates with down- Paul S, Mandal SK, Bhattacharyya SS, Boujedaini N,
regulation of cyclin D1 expression and CDK4-mediated Khuda-Bukhsh AR. 2010. In vitro and in vivo studies
retinoblastoma protein phosphorylation. Oncogene., demonstrate anticancer property of root extract of
21:8852-8861. Polygala senega. J Acupunct Meridian Stud., 3:188-196.
Nair V, Dai Z, Khan M, Ciolino HP. 2011. Phillips JM, Clark C, Herman-Ferdinandez L, Moore
Pomegranate extract induces cell cycle arrest and alters -Medlin T, Rong X, Gill JR,Clifford JL, Abreo F,
cellular phenotype of human pancreatic cancer cells, Nathan CO. 2011. Curcumin inhibits skin squamous
Anticancer Res 31:2699-2704. cell carcinoma tumor growth in vivo. Otolaryngol Head
Neck Surg., 145:58-63.
Naowaratwattana W, De-Eknamkul W, De Mejia EG.
2010. Phenolic-containing organic extracts of mulberry Preza AM, Jaramillo ME, Puebla AM, Mateos JC,
(Morus alba L.) leaves inhibit HepG2 hepatoma cells Hernndez R, Lugo E. 2010. Antitumor activity
through G2/M phase arrest, induction of apoptosis, and against murine lymphoma L5178Y model of proteins
inhibition of topoisomerase II activity. J Med Food., from cacao (Theobroma cacao L.) seeds in relation with
13:1045-1056. in vitro antioxidant activity. BMC Complement Altern
Med., 10:61.
Ngamkitidechakul C, Jaijoy K, Hansakul P,
Soonthornchareonnon N, Sireeratawong S. 2010. Qian H, Yang Y and Wang X. 2011. Curcumin
Antitumour effects of Phyllanthus emblica L.: induction enhanced adriamycin-induced human liver-derived
of cancer cell apoptosis and inhibition of in vivo tumour Hepatoma G2 cell death through activation of
promotion and in vitro invasion of human cancer cells. mitochondria-mediated apoptosis and autophagy. Eur J
Phytother Res., 24:1405-1413. Pharm Sci., 43:125-131.
Ning L, Wentworth L, Chen H, Weber SM. 2009. Rao J, Xu DR, Zheng FM, Long ZJ, Huang SS, Wu
Down-regulation of Notch1 signaling inhibits tumor X, Zhou WH, Huang RW, Liu Q. 2011. Curcumin
growth in human hepatocellular carcinoma. Am J Transl reduces expression of Bcl-2, leading to apoptosis in
Res., 1:358-366. daunorubicin-insensitive CD34+ acute myeloid leukemia
cell lines and primary sorted CD34+ acute myeloid
O'Sullivan-Coyne G, O'Sullivan GC, O'Donovan TR,
leukemia cells, J Transl Med., 19; 9:71.
Piwocka K, McKenna SL. 2009. Curcumin induces
apoptosis-independent death in oesophageal cancer cells. Rayanil KO, Bunchornmaspan P, Tuntiwachwuttikul
Br J Cancer., 101:1585-1595. P. 2011. A new phenolic compound with anticancer
activity from the wood of Millettia leucantha. O. Arch
Pandelidou M, Dimas K, Georgopoulos A,
Pharm Res., 34:881-886.
Hatziantoniou S, Demetzos C. 2011. Preparation and
characterization of lyophilised egg PC liposomes Rejinold NS, Muthunarayanan M, Chennazhi KP,
incorporating curcumin and evaluation of its activity Nair SV, Jayakumar R. 2011a. Curcumin loaded
against colorectal cancer cell lines, J Nanosci fibrinogen nanoparticles for cancer drug delivery. J
Nanotechnol., 11:1259-1266. Biomed Nanotechnol., 7:521-534.

Ghosh et al., 2012
Rejinold NS, Muthunarayanan M, Divyarani VV, Saini S, Arora S, Majid S, Shahryari V, Chen Y,
Sreerekha PR, Chennazhi KP, Nair SV, Tamura H, Deng G, Yamamura S, Ueno K, Dahiya R. 2011.
Jayakumar R. 2011b. Curcumin-loaded biocompatible Curcumin Modulates MicroRNA-203-Mediated
thermoresponsive polymeric nanoparticles for cancer Regulation of the Src-Akt Axis in Bladder Cancer.
drug delivery. J Colloid Interface Sci., 360:39-51. Cancer Prev Res (Phila)., 4:1698-1709.
Rejinold NS, Sreerekha PR, Chennazhi KP, Nair SV, Saxena A, Saxena AK, Singh J, Bhushan S. 2010.
Jayakumar R. 2011c. Biocompatible, biodegradable Natural antioxidants synergistically enhance the
an d th erm o-sen sit i ve chi t osan -g-pol y (N- anticancer potential of AP9-cd, a novel lignan
isopropylacrylamide) nanocarrier for curcumin drug composition from Cedrus deodara in human leukemia
delivery. Int J Biol Macromol., 49:161-172. HL-60 cells. Chem Biol Interact., 188:580-590.
Reuter S, Gupta SC, Park B, Goel A, Aggarwal BB. Saydmohammed M, Joseph D, Syed V. 2010.
2011. Epigenetic changes induced by curcumin and other Curcumin suppresses constitutive activation of STAT-3
natural compounds. Genes Nutr., 6:93-108. by up-regulating protein inhibitor of activated STAT-3
(PIAS-3) in ovarian and endometrial cancer cells. J Cell
Rowe DL, Ozbay T, O'Regan RM, Nahta R. 2009.
Biochem., 110:447-456.
Modulation of the BRCA1 Protein and Induction of
Apoptosis in Triple Negative Breast Cancer Cell Lines Schaaf C, Shan B, Buchfelder M, Losa M, Kreutzer J,
by the Polyphenolic Compound Curcumin. Breast Rachinger W, Stalla GK, Schilling T, Arzt E, Perone
Cancer (Auckl)., 3:61-75. MJ, Renner U. 2009. Curcumin acts as anti-tumorigenic
and hormone-suppressive agent in murine and human
Rungphanichkul N, Nimmannit U, Muangsiri W,
pituitary tumour cells in vitro and in vivo. Endocr Relat
Rojsitthisak P. 2011. Preparation of curcuminoid
Cancer., 16:1339-1350.
niosomes for enhancement of skin permeation.
Pharmazie., 66:570-575. Schaaf C, Shan B, Onofri C, Stalla GK, Arzt E,
Schilling T, Perone MJ, Renner U. 2010. Curcumin
Saha A, Kuzuhara T, Echigo N, Fujii A, Suganuma
inhibits the growth, induces apoptosis and modulates the
M, Fujiki H. 2010. Apoptosis of human lung cancer
function of pituitary folliculostellate cells.
cells by curcumin mediated through up-regulation of
Neuroendocrinology., 91:200-210.
"growth arrest and DNA damage inducible genes 45 and
153". Biol Pharm Bull 33:1291-1299. Selvaduray KR, Radhakrishnan AK, Kutty MK,
Nesaretnam K. 2010. Palm tocotrienols inhibit
Sahu A, Kasoju N, Goswami P, Bora U. 2011.
proliferation of murine mammary cancer cells and induce
Encapsulation of curcumin in Pluronic block copolymer
expression of interleukin-24 mRNA, J Interferon
micelles for drug delivery applications. J Biomater
Cytokine Res., 30:909-916.
Appl., 25:619-639.
Selvam P, El-Sherbiny IM, Smyth HD. 2011.
Sahu RP, Batra S, Srivastava SK. 2009. Activation of
Swellable hydrogel particles for controlled release
ATM/Chk1 by curcumin causes cell cycle arrest and
pulmonary administration using propellant-driven
apoptosis in human pancreatic cancer cells. Br. J.
metered dose inhalers. J Aerosol Med Pulm Drug Deliv.,
Cancer., 100:1425-1433.
24:25-34.

Ghosh et al., 2012
Selvaraj Nagarajan, Bo Siva Rami Reddy, Jhon cancer. Nutr Cancer., 63:161-173.
Tsibouklis. 2011. In vitro effect on cancer cells:
Shao J, Zheng D, Jiang Z, Xu H, Hu Y, Li X, Lu X.
Synthesis and preparation of polyurethane membranes
2011. Curcumin delivery by methoxy polyethylene
for controlled delivery of curcumin Journal of
glycol-poly(caprolactone) nanoparticles inhibits the
biomedical materials research. Part A 12 99:410-417.
growth of C6 glioma cells. Acta Biochim Biophys Sin
Sen GS, Mohanty S, Hossain DM, Bhattacharyya S, (Shanghai)., 43:267-274.
Banerjee S, Chakraborty J, Saha S, Ray P,
Sharma RA, Gescher AJ, Steward WP. 2005.
Bhattacharjee P, Mandal D, Bhattacharya A,
Curcumin: the story so far. Eur J Cancer., 41:1955-1968.
Chattopadhyay S, Das T, Sa G. 2011. Curcumin
enhances the efficacy of chemotherapy by tailoring Shelma R, Sharma CP. 2011. Submicroparticles
p65NF{kappa}B-p300 cross-talk in favor of p53-p300 in composed of amphiphilic chitosan derivative for oral
breast cancer. J Biol Chem., 286:42232-42247. insulin and curcumin release applications. Colloids Surf
B Biointerfaces., 88:722-728.
Seo JH, Jeong KJ, Oh WJ, Sul HJ, Sohn JS, Kim YK,
Cho do Y, Kang JK, Park CG, Lee HY. 2010. Shimizu M, Adachi S, Masuda M, Kozawa O,
Lysophosphatidic acid induces STAT3 phosphorylation Moriwaki H. 2011. Cancer chemoprevention with green
and ovarian cancer cell motility: their inhibition by tea catechins by targeting receptor tyrosine kinases. Mol
curcumin. Cancer Lett., 288:50-56. Nutr Food Res., 55:832-843.
Sertel S, Eichhorn T, Plinkert PK, Efferth T. 2011. Shrikanth Reddy, Bharat B. Aggarwal. 1994.
Anticancer activity of Salvia officinalis essential oil Curcumin is a non-competitive and selective inhibitor of
against HNSCC cell line (UMSCC1). [Article in phosphorylase kinase, FEBS Letters., 341:19-22.
German] HNO.
Shu L, Khor TO, Lee JH, Boyanapalli SS, Huang Y,
Setthacheewakul S, Kedjinda W, Maneenuan D, Wu TY, Saw CL, Cheung KL, Kong AN. 2011.
Wiwattanapatapee R. 2011. Controlled release of oral Epigenetic CpG Demethylation of the Promoter and
tetrahydrocurcumin from a novel self-emulsifying Reactivation of the Expression of Neurog1 by Curcumin
floating drug delivery system (SEFDDS). AAPS in Prostate LNCaP Cells. AAPS J., Sep 22.
PharmSciTech., 12:152-164.
Singh AT, Ghosh M, Forte TM, Ryan RO, Gordon
Shahani K, Swaminathan SK, Freeman D, Blum A, Li. 2011. Curcumin nanodisk-induced apoptosis in
Ma L, Panyam J. 2010. Injectable sustained release mantle cell lymphoma. Leuk Lymphoma., 52:1537-1543.
microparticles of curcumin: a new concept for cancer
Singh M, Singh R, Bhui K, Tyagi S, Mahmood Z,
chemoprevention. Cancer Res., 70:4443-4452.
Shukla Y. 2011. Tea polyphenols induce apoptosis
Shalaby E. 2011. Algae as promising organisms for through mitochondrial pathway and by inhibiting nuclear
environment and health. Plant Signal Behav., 6. factor-kappaB and Akt activation in human cervical
cancer cells. Oncol Res., 9:245-257.
Shanmugam MK, Kannaiyan R, Sethi G. 2011.
Targeting cell signaling and apoptotic pathways by Singh S, Aggarwal BB. 1995. Activation of
dietary agents: role in the prevention and treatment of transcription factor NF-kappa B is suppressed by

Ghosh et al., 2012
curcumin (diferuloylmethane). J Biol Chem., 270:24995- Thamake SI, Raut SL, Ranjan AP, Gryczynski Z,
25000. Vishwanatha JK. 2011. Surface functionalization of
PLGA nanoparticles by non-covalent insertion of a homo
Spiller SE, Logsdon NJ, Deckard LA, Sontheimer H.
-bifunctional spacer for active targeting in cancer
2011. Inhibition of nuclear factor kappa-B signaling
therapy. Nanotechnology., 22:035101.
reduces growth in medulloblastoma in vivo. BMC
Cancer., 11:136 Thangapazham RL, Puri A, Tele S, Blumenthal R,
Maheshwari RK. 2008. Evaluation of a nanotechnology
Sreelatha S, Jeyachitra A, Padma PR. 2011.
-based carrier for delivery of curcumin in prostate cancer
Antiproliferation and induction of apoptosis by Moringa
cells. Int J Oncol., 32:1119-1123.
oleifera leaf extract on human cancer cells. Food Chem
Toxicol., 49:1270-1275. Tian F, Song M, Xu PR, Liu HT, Xue LX. 2008.
Curcumin promotes apoptosis of esophageal squamous
Srimal RC, Dhawan BN. 1973. Pharmacology of
carcinoma cell lines through inhibition of NF-kappaB
diferuloyl methane (curcumin), a non-steroidal anti-
signaling pathway. [Article in Chinese]. Ai Zheng.,
inflammatory agent. J Pharm Pharmacol., 25:447-452.
27:566-570.
Syng-Ai C, Kumari AL, Khar A. 2004. Effect of
Tripathi A, Puddick J, Prinsep MR, Rottmann M,
curcumin on normal and tumor cells: role of glutathione
Chan KP, Chen DY, Tan LT. 2011. Lagunamide C, a
and bcl-2. Mol Cancer Ther., 3:1101-1108.
cytotoxic cyclodepsipeptide from the marine
Szliszka E, Krol W. 2011. Soy isoflavones augment the cyanobacterium Lyngbya majuscula., Phytochemistry
effect of TRAIL-mediated apoptotic death in prostate 72:2369-2375.
cancer cells. Oncol Rep., 26:533-541.
Tsuiji K, Takeda T, Li B, Wakabayashi A, Kondo A,
Tadmor T, Polliack A. 2011. Mantle cell lymphoma: Kimura T, Yaegashi N. 2011. Inhibitory effect of
curcumin nanodisks and possible new concepts on drug curcumin on uterine leiomyoma cell proliferation.
delivery for an incurable lymphoma. Leuk Lymphoma., Gynecol Endocrinol., 27:512-517.
52:1418-1420.
Tuntiwechapikul W, Taka T, Songsomboon C,
Takeda S, Matsuo K, Yaji K, Okajima-Miyazaki S, Kaewtunjai N, Imsumran A, Makonkawkeyoon L,
Harada M, Miyoshi H, Okamoto Y, Amamoto T, Pompimon W, and Lee TR. 2010. Ginger extract
Shindo M, Omiecinski CJ, Aramaki H. 2011. (--)- inhibits human telomerase reverse transcriptase and c-
Xanthatin selectively induces GADD45 and stimulates Myc expression in A549 lung cancer cells. J Med Food.,
caspase-independent cell death in human breast cancer 13:1347-1354.
MDA-MB-231 cells. Chem Res Toxicol., 24:855-865.
Ulbricht CE, Chao W. 2010. Review. Phytochemicals
Tang XQ, Bi H, Feng JQ, Cao JG. 2005. Effect of in the oncology setting. Curr Treat Options Oncol., 11:95
curcumin on multidrug resistance in resistant human -106.
gastric carcinoma cell line SGC7901/VCR. Acta
Vilas-Zornoza A, Agirre X, Martn-Palanco V,
Pharmacol Sin., 26:1009-1016.
Martn-Subero JI, San Jos-Eneriz E, Garate L,
lvarez S, Miranda E, Rodrguez-Otero P, Rifn J,

Ghosh et al., 2012
Torres A, Calasanz MJ, Cruz Cigudosa J, Romn- Wang X, Xia X, Xu C, Xu J, Wang P, Xiang J, Bai D,
Gmez J, Prsper F. 2011. Frequent and simultaneous Leung AW. 2011b. Ultrasound-induced cell death of
epigenetic inactivation of TP53 pathway genes in acute nasopharyngeal carcinoma cells in the presence of
lymphoblastic leukemia. PLoS One., 6:e17012. curcumin. Integr Cancer Ther., 10:70-76.
Vishvakarma NK, Kumar A, Singh SM. 2011. Role of Wang XN, Bashyal BP, Wijeratne EM, U'ren JM, Liu
cur cumin -dependen t m odul at i on of t um or MX, Gunatilaka MK, Arnold AE, Gunatilaka AA.
microenvironment of a murine T cell lymphoma in 2011. Smardaesidins A-G, Isopimarane and 20-nor-
altered regulation of tumor cell survival. Toxicol Appl Isopimarane Diterpenoids from Smardaea sp., a Fungal
Pharmacol., 252:298-306. Endophyte of the Moss Ceratodon purpureus (1) J Nat
Prod., 74:2052-2061.
Vu HA, Beppu Y, Chi HT, Sasaki K, Yamamoto H,
Xinh PT, Tanii T, Hara Y, Watanabe T, Sato Y, Watson JL, Greenshields A, Hill R, Hilchie A, Lee
Ohdomari. I. 2010. Green tea epigallocatechin gallate PW, Giacomantonio CA, Hoskin DW. 2010. Curcumin
exhibits anticancer effect in human pancreatic carcinoma -induced apoptosis in ovarian carcinoma cells is p53-
cells via the inhibition of both focal adhesion kinase and independent and involves p38 mitogen-activated protein
insulin-like growth factor-I receptor, J Biomed kinase activation and downregulation of Bcl-2 and
Biotechnol., 2010:290516. survivin expression and Akt signaling. Mol Carcinog.,
49:13-24.
Wahl O, Oswald M, Tretzel L, Herres E, Arend J,
Efferth T. 2011. Inhibition of tumor angiogenesis by Wei CC, Ball S, Lin L, Liu A, Fuchs JR, Li PK, Li C,
antibodies, synthetic small molecules and natural Lin J. 2011. Two small molecule compounds, LLL12
products, Curr Med Chem., 18:3136-3155. and FLLL32, exhibit potent inhibitory activity on
STAT3 in human rhabdomyosarcoma cells. Int J Oncol.,
Wang C, Henkes LM, Doughty LB, He M, Wang D,
38:279-285.
Meyer-Almes FJ, Cheng YQ. 2011. Thailandepsins:
bacterial products with potent histone deacetylase Wong TS, Chan WS, Li CH, Liu RW, Tang WW,
inhibitory activities and broad-spectrum antiproliferative Tsao SW, Tsang RK, Ho WK, Wei WI, Chan JY.
activities. J Nat Prod., 74:2031-2038. 2010. Curcumin alters the migratory phenotype of
nasopharyngeal carcinoma cells through up-regulation of
Wang O, Liu S, Zou J, Lu L, Chen L, Qiu S, Li H, Lu
E-cadherin. Anticancer Res., 30(7):2851-2856.
X. 2011. Anticancer activity of 2, 3, 19, 23-
Tetrahydroxyurs-12-en-28-oic acid (THA), a novel Wu SH, Hang LW, Yang JS, Chen HY, Lin HY,
triterpenoid isolated from Sinojackia sarcocarpa. PLoS Chiang JH, Lu CC, Yang JL, Lai TY, Ko YC, Chung
One., 6:e21130. JG. 2010. Curcumin induces apoptosis in human non-
small cell lung cancer NCI-H460 cells through ER stress
Wang X, Xia X, Leung AW, Xiang J, Jiang Y, Wang
and caspase cascade- and mitochondria-dependent
P, Xu J, Yu H, Bai D, Xu C. 2011a. Ultrasound induces
pathways. Anticancer Res., 30:2125-2133.
cellular destruction of nasopharyngeal carcinoma cells in
the presence of curcumin. Ultrasonics., 51:165-170. Xiao D, Zeng Y, Prakash L, Badmaev V, Majeed M,
and Singh SV. 2011. Reactive oxygen species-
dependent apoptosis by gugulipid extract of Ayurvedic
Ghosh et al., 2012
medicine plant Commiphora mukul in human prostate H446 Cell Apoptosis via the Reactive Oxygen Species-
cancer cells is regulated by c-Jun N-terminal kinase. Mol Mediated Mitochondrial Pathway and Not the Cell Death
Pharmacol., 79:499-507. Receptor Pathway, DNA Cell Biol., Jun 28.
Xiao H, Xiao Q, Zhang K, Zuo X, Shrestha UK. 2010. Yin H, Guo R, Xu Y, Zheng Y, Hou Z, Dai X, Zhang
Reversal of multidrug resistance by curcumin through Z, Zheng D, Xu H. 2011. Synergistic antitumor
FA/BRCA pathway in multiple myeloma cell line MOLP efficiency of docetaxel and curcumin against lung
-2/R. Ann Hematol., 89:399-404. cancer. Acta Biochim Biophys Sin (Shanghai).
Xie X, Tao Q, Zou Y, Zhang F, Guo M, Wang Y, Yu H, Huang Q. 2011. Investigation of the absorption
Wang H, Zhou Q, Yu S. 2011. PLGA nanoparticles mechanism of solubilized curcumin using Caco-2 cell
improve the oral bioavailability of curcumin in rats: monolayers. J Agric Food Chem 59(17):9120-9126,
characterizations and mechanisms J Agric Food Chem., 2011.
59:9280-9289.
Yu LL, Wu JG, Dai N, Yu HG and Si JM. 2011.
Yadav VS, Mishra KP, and Singh DP. 2010. Curcumin Curcumin reverses chemoresistance of human gastric
inhibits Jurkat cell proliferation by inducing apoptosis cancer cells by downregulating the NF-B transcription
via activation-induced cell death. Biomed Pharmacother., factor. Oncol Rep., 26:1197-1203.
Sep 20.
Zanotto-Filho A, Braganhol E, Edelweiss MI, Behr
Yallapu MM, Dobberpuhl MR, Maher DM, Jaggi M, GA, Zanin R, Schrder R, Simes-Pires A, Battastini
Chauhan SC. 2011a. Design of Curcumin loaded AM, Moreira JC. 2011. The curry spice curcumin
Cellulose Nanoparticles for Prostate Cancer. Curr Drug selectively inhibits cancer cells growth in vitro and in
Metab., Sep 5. preclinical model of glioblastoma. J Nutr Biochem.,
Yallapu MM, Gupta BK, Jaggi M, Chauhan SC. Zhang C, Li B, Zhang X, Hazarika P, Aggarwal BB,
2010a. Fabrication of curcumin encapsulated PLGA Duvic M. 2010. Curcumin selectively induces apoptosis
nanoparticles for improved therapeutic effects in in cutaneous T-cell lymphoma cell lines and patients'
metastatic cancer cells. J Colloid Interface Sci.,351:19- PBMCs: potential role for STAT-3 and NF-kappaB
29. signaling. J Invest Dermatol., 130:2110-2119.
Yallapu MM, Jaggi M, Chauhan SC. 2011b. Zhang H, Zhang L, Yuan P, Wang C. 2011.
Curcumin nanoformulations: a future nanomedicine for Preparation and in vitro release characteristics of
cancer. Drug Discov Today., Sep 18. curcumin in nanosuspensions, Zhongguo Zhong Yao Za
Zhi., 36:132-135.
Yallapu MM, Jaggi M, Chauhan SC. 2010b. Poly (-
cyclodextrin)/curcumin self-assembly: a novel approach Zhang J, Du Y, Wu C, Ren X, Ti X, Shi J, Zhao F,
to improve curcumin delivery and its therapeutic efficacy Yin H. 2010. Curcumin promotes apoptosis in human
in prostate cancer cells. Macromol Biosci., 10:1141- lung adenocarcinoma cells through miR-186* signaling
1151. pathway. Oncol Rep., 24:1217-1223.
Yang CL, Ma YG, Xue YX, Liu YY, Xie H, Qiu GR. Zhang L, Cao F, Ding B, Li Q, Xi Y, Zhai G. 2011.
2011. Curcumin Induces Small Cell Lung Cancer NCI- Eudragit S100 coated calcium pectinate microspheres

Ghosh et al., 2012
of curcumin for colon targeting. J Microencapsul.,

28:659-667.
Zhang Q, Zhong Y, Yan LN, Sun X, Gong T, Zhang

ZR. 2011. Synthesis and preliminary evaluation of
curcumin analogues as cytotoxic agents. Bioorg Med
Chem Lett., 21:1010-1014.
Zhao LX, Huang SX, Tang SK, Jiang CL, Duan Y,

Beutler JA, Henrich CJ, McMahon JB, Schmid T,
Blees JS, Colburn NH, Rajski SR, Shen B. 2011.
Actinopolysporins A-C and tubercidin as a Pdcd4
stabilizer from the halophilic actinomycete
Actinopolyspora erythraea YIM 90600. J Nat Prod.,
23;74:1990-1995.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. Xiao H,

Zhang KJ. 2008. Antiproliferative effect of curcumin
combined with cyclophosmide on the growth of human
lymphoma cell line HT/CTX with drug resistance and its
relation with FA/BRCA pathway. [Article in Chinese] J
Experimental hematology., 16:804-808.
Zong H, Wang F, Fan QX, and Wang LX. 2011.

Curcumin inhibits metastatic progression of breast
cancer cell through suppression of urokinase-type
plasminogen activator by NF-kappa B signaling
pathways. Mol Biol Rep.

Advantages
Affordable Charges
Quick processing
Extensive indexing

Journal of Research in Biology - Volume 2 Issue 3

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Journal of Research in Biology - Volume 2 Issue 3

Uploaded by

Copyright:

Available Formats

An International Online Open Access

Acute toxicity of insecticide, Diazinon and fungicide, Tilt (Propiconazole) on Pacific

Web Address: Article Citation:

160-166 | JRB | 2012 | Vol 2 | No 3

INTRODUCTION the household environment (Cox, 1992). Trade names for

Table1. Acute toxicity of Diazinon in 96 h on Table2. Acute toxicity of Tilt in 96 h on