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Authors: ABSTRACT:
Ihejirika CE1, Ogbulie JN2,
Nwabueze RN2, Orji JC2, The occurrence and population of pathogenic bacteria at certain points of
Ihejirika OC3, Adieze IE2, Imo River, were determined. Water samples were collected for two seasons at major
Azubike OC4 and Ibe IJ5.
points of human activities along the river and subjected to standard microbiological
and statistical analyses. Total heterotrophic bacterial count (2.9 x 10 9 3.7 x 103 CFU/
ml) and total coliform bacterial counts (9.0 x 106 2.5 x 102 CFU/ml) showed
Institution: significant variation at P<0.05 for the two seasons. Percentage occurrence of
1
Department of individual isolates across the sampling points showed the presence of Escherichia coli
Environmental Technology, (100%), Klebsiella spp. (71.0%), Shigella spp. (71.0%), Salmonella spp.(71.0%), Proteus
2 spp.(42.9%), Vibro spp. (42.9%), Pseudomonas spp. (42.9%), Staphylococcus spp.
Department of
Microbiology, (85.7%), Bacillus spp. (100%), Enterobacter spp. (57.1%), Citrobacter spp. (14.3%),
3
Department of Public Serratia spp.(14.3%) and Streptococcus spp.(14.3%). These organisms are of public
Health and health importance and imply that Imo River should be protected from pollution to
4
Department of avoid possible diseases outbreak and transmission.
Biotechnology, Federal
University of Technology,
Owerri,
5
Department of Biology and Keywords:
Microbiology, Federal Imo River, pathogenic bacteria, percentage occurrence, diseases outbreak
Polytechnic Nekede, Owerri. and transmission.
Web Address:
http://jresearchbiology.com/ Ficus Publishers.
Documents/RA0039.pdf.
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hours. The samples were collected in two seasons locations of the Imo River is as shown in Table
dry and rainy seasons for two years. The dry 1.The THBC during the dry season ranged from 3.6
season was between November and March while x 103 -1.23 x 106cfu/ml while the TCBC during the
the rainy season was between May and September. dry season ranged from 1.65 x 103 8.7 x 106 cfu/
Microbiological analysis ml. These results were above the limits of EPA
Sterilization of media was carried out by Maximum Contaminant Levels (MCLS) of
moist heat sterilization method using autoclave at <100cfu/ml in drinking water (USEPA, 2003).
1210C, 15psi for 15 minutes. Heat stable materials Table 2 shows the % occurrencea of
were sterilized using hot air oven at 1600C for 1 bacterial isolates from the different sampling point
hour as described by Cruickshank et al. (1982). or location of Imo River. A total of 14 organisms
Heat labile materials were aseptically rinsed with were isolated from the different locations. These
alcohol and distilled water. The water samples organisms include Escherichia coli (100%),
were aseptically subjected to 10 fold serial dilutions Klebsiella spp (71.0%), Shigella spp (71.0%),
to dilute the population of microorganism Salmonella spp (71.0%), Proteus spp (42.9%),
sufficiently in sterile blanks of 9ml peptone water Vibro spp(42.9%), Pseudomonas spp(85.7%),
and then plated to produce discrete colonies for Staphylococcus spp (85.7%), Bacillus spp
easy enumeration. The media used include Nutrient (100.0%), Enterobacter spp (57.1%), Citrobacter
agar, MacConkey agar, Eosin Methylene Blue agar, spp (14.3%), Serratia spp (14.3%) and
TCBS, and Salmonella Shigella agar. All media Streptococcus spp (14.3%). This implied that
were prepared as directed by the manufacturer. The Escherichia coli and Bacillus spp were isolated
method was adopted for the inoculation of media. from all the 14 sampling points, Pseudomonas spp
Spread plates of appropriately diluted samples were and Staphylococcus spp isolated from 6 out of the 7
incubated at 370C for 24 hours for heterotrophic sampling points and so on.
bacterial count (THBC) while total coliform The % occurrenceb of total bacterial isolates
bacterial count (TCBC) were determined after at each sampling point showed Owerrinta (69.2%),
incubation at 450C for 24 hours in MacConkey Udo (61.5%), Alulu (61.5%), Akwette (38.4%),
agar. Identification of isolates was based on the Ekenobizi (76.9%), Owaza (53.8%), and Obigbo
scheme described by Cheesborough (1984). (53.8%).
The results were subjected to Analysis of Variance Table 3 shows the result of seasonal
(ANOVA) by using Statistical Program for Social variation of THBC and TCBC of Ekenobizi Point
Sciences (SPSS) and percentage occurrence. of Imo River. There were significant seasonal
variations (P<0.05) in THBC at P = 0.00 and T-
RESULTS AND DISCUSSION: 31.92. The mean THBC values were 1.2 x 106 cfu/
Results: ml (Dry) and 5.0 x 105 cfu/ml (Rainy). There were
The result of seasonal variation of Total no significant seasonal variations in TCBC at P =
Heterotrophic Bacterial Count (THBC) and Total 0.197 and T= 1.54. The mean TCBC values were
Coliform Bacterial Count (TCBC) at all sampling 8.7 x 105 cfu/ml (Dry) and 3.7 x 104 cfu/ml (Rainy).
Table 2: Percentage occurrence of microbial isolates at different sampling points of Imo River
Table 4 shows the result of seasonal Imo River. There were significant seasonal
variation of THBC and TCBC of Udo point of Imo variations in THBC at P = 0.00 and T = 85.99. The
River. There were significant seasonal variations in mean THBC values were 8.6 x103cfu/ml (Dry) and
THBC at P = 0.00 and T-99.98. The mean THBC 1.3 x 107 cfu/ml (Rainy). There were significant
values were 2.0 x 105 cfu/ml (Dry) and 2.0 x 109 seasonal variations in TCBC at P = 0.00 and T=
cfu/ml (Rainy). There were significant seasonal 32.70. The mean TCBC values were 5.0 x 103cfu/
variations in TCBC at P = 0.00 and T= 11.85. The ml (Dry) and 1.0 x 106cfu/ml (Rainy).
mean TCBC values were 1.9 x 104 cfu/ml (Dry) and Table 7 shows the result of seasonal
8.9 x 106 cfu/ml (Rainy). variation of THBC and TCBC of Owaza Point of
Table 5 shows the result of seasonal Imo River. There were significant seasonal
variation of THBC and TCBC of Owerrinta Point variations in THBC at P = 0.01 and T = 4.47. The
of Imo River. There were significant seasonal mean THBC values were 3.7 x103cfu/ml (Dry) and
variations in THBC at P = 0.00 and T = 32.04. The 4.1 x 103cfu/ml (Rainy). There were significant
mean THBC values were 1.8 x 104cfu/ml (Dry) and seasonal variations in TCBC at P = 0.00 and T=
5.5 x 104 cfu/ml (Rainy). There were significant 23.35. The mean TCBC values were 1.7 x 103cfu/
seasonal variations in TCBC at P = 0.00 and T= ml (Dry) and 2.5 x 102cfu/ml (Rainy).
60.21. The mean TCBC values were 4.8 x 103cfu/ Table 8 shows the result of seasonal
ml (Dry) and 2.0 x 104cfu/ml (Rainy). variation of THBC and TCBC of Obigbo Point of
Table 6 shows the result of seasonal Imo River. There were significant seasonal
variation of THBC and TCBC of Alulu Point of variations in THBC at P = 0.00 and T = 44.71. The
Table 3: Seasonal variation of THBC and TCBC of Ekenobizi Point of Imo River.
Table 4: Seasonal variation of THBC and TCBC of Udo Point of Imo River.
Table 5: Seasonal variation of THBC and TCBC of Owerrinta Point of Imo River.
Table 6: Seasonal variation of THBC and TCBC of Alulu Point of Imo River.
mean THBC values were 4.5 x104cfu/ml (Dry) and water (USEPA, 2003). The variation in the dry and
2.9 x 106cfu/ml (Rainy). There were significant rainy seasons might be due to heavy rainfalls
seasonal variations in TCBC at P = 0.00 and T= resulting to sewage overflow and surface runoff.
18.30. The mean TCBC values were 1.9 x 104cfu/ This was demonstrated by previous studies (Goyal
ml (Dry) and 3.0 x 103cfu/ml (Rainy) et al., 1977). These results suggest that heavy
Table 9 shows the result of seasonal rainfall events may contribute to Imo River
variation of THBC and TCBC of Akwette Point of contamination and may give rise to increased
Imo River. There were significant seasonal concentration of pathogens in the water.
variations in THBC at P = 0.00 and T 26.47. The The coliform test is a reliable indicator of
mean THBC values were 4.1 x104cfu/ml (Dry) and the possible presence of fecal contamination and is,
5.6 x 106cfu/ml (Rainy). There were significant consequently, correlated with pathogens. The
seasonal variations in TCBC at P = 0.00 and T= USEPA MCL is less than one coliform per 100ml
24.49. The mean TCBC values were 1.0 x 104cfu/ (USEPA, 2003).
ml (Dry) and 5.0 x 104cfu/ml (Rainy). The Total Heterotrophic Bacteria Count
(THBC) test also called total count or plate
DISCUSSION: count, provides an estimate of the total number of
The result of Total Heterotrophic Bacterial bacteria in a sample that will develop into colonies
Count (THBC) and Total Coliform Bacterial Count during a period of incubation in a nutrient. This test
(TCBC) as shown in table 1 were above the USEPA detects a broad group of bacteria including
Maximum Contaminant Levels (MCLS) in drinking pathogens, and opportunistic pathogens, but it does
Table 7: Seasonal variation of THBC and TCBC of Owaza Point of Imo River.
not pretend to report all of the bacteria in the water points. This result is supported by the works of
sample examined. High THBC may be an indicator Health Canada (2006) and Cabral (2010). E. coli is
of poor general biological quality of drinking water a coliform bacterium found exclusively in the
(USEPA, 2003). There might be presence of digestive tract of warm blooded animals, including
emerging waterborne pathogenic bacteria like humans. This might be responsible for the highest
Mycobacterium avium, Legionella spp., percentage occurrence (Swerdlow et al., 1992). As
Helicobacter spp., and Aeromonas hydrophyla in such E. coli is used in the drinking water industry as
Imo River that might not be isolated through the definitive indicator of recent fecal
conventional laboratory techniques. Health contamination of water.
agencies like the USEPA and World Health Citrobacter spp, Klebsiella spp, Salmonella,
Organization (WHO) have avoided setting Shigella, and Serratia spp are present in most
standards for plate counts possibly for lack of individuals although in low numbers, while Proteus
pathogenicity and great variation in density, spp and Enterobacter spp. are only present in
encountered (Dezuane, 1990). A recommended minority of humans (Wilson, 2005). Therefore,
MCL for human drinking water has not yet been these organisms are not suitable as indicators of
proposed, but the USEPA does recognize the water fecal pollution of the environment. This might be
quality deterioration implied by high plate counts. responsible for their <100% distribution in the
The upper limit for portable water is usually waters. This is supported by the work of Cabral
500cfu/ml. Dezuane (1990) says that water with (2010).While most strains of E. coli are non-
counts under 100cfu/ml should be considered pathogenic, some can cause serious diarrheal
portable and values 100 -500/ml is infections in human (Health Canada, 2006), urinary
questionable. Therefore Imo River samples have tract infections (Scheatz and Strockbin, 2005), and
questionable water quality. distribution of erythrocytes (Moe,
Among these organisms, the members of the 1997).Citrobacter spp (14.3%) is included in a
family Enterobacteriaceae include Escherichia number of pathogenic bacteria capable of causing
coli, Citrobacter spp., Klebsiella spp, Proteus spp, serious disease and being discharged into rivers
Enterobacter spp, Salmonella spp, Shigella spp, and (Donovan et al.,2008); has ability to produce an
Serratia spp (Prescott et al., 2005). As E. coli was enterotoxin and this become an intestinal pathogen
isolated from all the sampling points, it indicated in environments such as water, sewage, soil and
recent fecal contamination of the different sampling food (Frederisksen and Sogaard, 2003).
Table 8: Seasonal variation of THBC and TCBC of Obigbo Point of Imo River.
Table 9: Seasonal variation of THBC and TCBC of Akwette Point of Imo River.
The presence of Citrobacter spp. is The association of Shigella spp (71.0%) with
significant since the species - C. freundii can cause some points of Imo River is an implication of the
meningitis with high morbidity and mortality fecal contamination of the River. This is in
(Donovan et al., 2008). agreement with the reports and works of WHO
Klebsiella spp (71.0%) are ubiquitous in the (2008) and Kapperud et al., (1995). That Shigella
environment. They have been found in a variety of spp with <100% occurrence at sampling points of
environmental situations, such as soil, vegetation, Imo River might imply its presence at some points
or water, and they influences, many biochemical, but are viable and non-culturable. This argument is
and geochemical processes (Cabral, 2010). They supported by the report of Faruque et al. (2002).The
have been recovered from aquatic environments implication of the presence of Shigella spp in Imo
receiving industrial wastewaters, plant products, River samples is the risk of possible outbreak of
fresh vegetables, food with a high content sugars shigellosis. This was in agreement with the report
and acids, frozen orange juice concentrate, of Emch et al. (2008).
sugarcane waste and living trees (Grimont et al., Enterobacter spp (57.1%) might be an
2005). It is because Klebsiella spp. has high % implication of fecal contamination at Imo River.
occurrence (71.0%) in Imo River and especially its This was supported by the works of Grimont and
isolation from Owerrinta Point of Imo River that Grimont (2005). Apart from fecal contamination
receives effluents from paper mill industries. Enterobacter spp might have been introduced from
Klebsiella spp can cause human diseases, ranging other sources like soil, polluted water, and plants.
from asymptomatic colonization of the intestinal, The implication of this ubiquity is that Enterobacter
urinary, or respiratory tract to fatal septicemia. spp cannot be used as an indicator of fecal
Klebsiella are mostly considered nosocomial contamination of water bodies. The presence of
pathogens (Grimont et al., 2005). Enterobacter spp in some samples of Imo River
The presence of Salmonella spp in Imo implied possible risk of outbreak of nosocomial
River samples might be due to contamination from infections such as urinary tract infections and other
municipal sewage agricultural pollution, and storm healthcare- associated infections. This argument is
water runoffs. This argument is supported by the supported by the reports of Hirdron et al. (2008).
reports of WHO (2008) and Arvanitidov et al. Proteus spp (42.9%) is an enteric pathogen
(2005). Though Salmonella spp can survive in associated with the feces of animals including
water bodies, its presence and multiplication can be humans. Its low % occurrence (42.9%) might be
influenced by seasonal conditions, temperature, because it exists in minority of human feces. This is
humidity, and pH, which might be contributory to supported by the reports of Wilson (2005).
the <100% (71.1%) occurrence of Salmonella spp Serratia spp (14.3%) is another enteric
in Imo River water samples. This is supported by pathogen associated with feces of some humans. Its
the work of Le Minor (2003). Salmonella spp are lower % occurrence might be due to low presence
responsible for two types of salmonellosis: (1) in feces of humans and subsequently its lower
Typhoid and paratyphoid fever; (2) gastroenteritis availability as a water borne pathogen. This is
(Le Minor, 2003). This implies that controlled supported by the report of Cabral (2010).
water sewage systems, pasteurization of foods and Vibro spp (42.9%) might be present in some
personal hygiene will reduce the incidence of samples due to contamination from birds, frogs,
typhoid fever (Popoff et al., 2005) that might result fishes and shell fish present in aquatic
from the use of Imo River. environments. This argument is supported by the
reports of Ali et al. (2001). They detected low for domestic and recreational activities.
distribution of Vibrio spp (42.9%) might be due to
environmental stress caused by adverse CONCLUSION:
environmental conditions making some of the cells Seasonal influences in addition to
viable but non-culturable, though retaining the anthropogenic activities contributed to the
potential for pathogenicity for significant periods of occurrence of water borne pathogens with high
time. This is in agreement with the report of Alam bacteria counts above established standards. The
et al. (2006) and Chaiyana et al. (2001). Vibrio spp River therefore is a source organism of public
especially V. cholerae is responsible for the disease health importance and should be protected from
cholera in humans (Cabral, 2010). human contamination and properly treated to avoid
Streptococcus spp (14.3%) were isolated in consequences.
one out of the seven locations and might be due to
fecal contamination of the point of Imo River. This REFERENCES:
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Monitoring of waterborne pathogens in water in
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Environ Microbiol., 74(7):2069-2078.
Authors: ABSTRACT:
Ganapathy Selvam G1,
Baskaran R2 and
Mohan PM3.
The role of cyanobacteria in distilleries effluent was studied in Kanchipuram
Tamilnadu, India. Totally 12 species of cyanobacteria belonging to 6 genera falling
Institution:
1
Department of under 4 families were identified. Six different genera of bacteria and 3 different
Microbiology, A.V.V.M, genera of fungi were isolated and identified. Among the cyanobacteria isolated,
Sri Pushpam College Nostoc muscorum was selected to treat the effluent. Distilleries effluent was the
(Autonomous), Thanjavur, potential source of cyanobacteria. Nostoc muscorum was found to be the most
Tamil Nadu, dominated genus in this effluent (heterocyst organism). The inoculation of Nostoc
India. muscorum resulted in removal of various chemicals as well as nutrients such as
nitrogen, ammonia and phosphorus from the effluent. It is concluded that Nostoc
2 and 3
Department of Ocean muscorum, a cyanobacteria found in the distilleries effluent, could be potentially
Studies and Marine Biology, employed for the treatment of distilleries effluent.
Pondicherry
University, Port Blair,
Andaman and Nicobar Island
-744 103, India.
Keywords:
Corresponding author: Distillery effluent, Microbial diversity, Bioremediation, Physico-Chemical
Baskaran R. analysis.
study the interaction of Cyanobacterium with the for the estimation of biochemical compound
distilleries effluent. Effluent uninoculated was variation in effluent treated and control culture.
taken as, control for physicochemical analysis (C).
BG11 medium inoculated with Nostoc muscorum for RESULTS AND DISCUSSION
the estimation of growth (CO). Inoculation was Isolation of cyanobacteria:
made by adding 3.0 ml uniform suspension of In the present investigations totally 12
Nostoc muscorum separately (0.03 mg ml-1). The species (Fig-1 and Table -1) of cyanobacteria
experiment was conducted under controlled distributed in five genera falling under four
conditions (Temperature 25 2oC with light different families were identified from the
intensities of 2500 lax provided form overhead cool distilleries effluent polluted habitat. Among the
with fluorescent tubes) for one month. Since the genera, Oscillatoria with six species followed by
growth of Nostoc muscorum was very slow in the Phormidium two species, Lyngbya, Microcystis,
effluent, a long duration was provided to get a Nostoc and Plectonema with single species each.
culture of exponential growth. The cultures were This is attributed to favorable conditions of
harvested on every 5th day by filtration through oxidizable organic matter, less DO and high
filter paper and washed repeatedly with distilled calcium content; (Table 4) an observation which
water. The filtered effluents (inoculated and supports Venkateswaralu (1969) who observed that
control) were used for physicochemical analysis. high orthophosphate levels favored the
Nostoc muscorum obtained both from the medium development of cyanobacterial bloom. Similar
and effluents were used for the estimation of observations were made in the present study with
growth (estimation of chlorophyll a). reference to various nutrients (Table-4).
Estimation of Chlorophyll a Heterocystous cyanobacteria have not been
The cultures were centrifuged at 5,000 x g recorded in polluted waters rich in nitrogen
for 10 minutes. The pellets were washed with (Boominathan et al., 2007). In contrary to that
distilled water; suspended in 80 per cent methanol present study heterocystous cyanobacteria have
and vortexed thoroughly. Then the tubes were already been recorded and is because of the
covered with aluminium foil to prevent solvent presence of lower concentration of various forms of
evaporation and incubated in a water bath set at nitrogen effluent as suggested by Vijayakumar et
60oC for 1 hr, in dark with occasional shaking. al., (2007).
After 1 hr the contents were cooled and centrifuged Isolation of fungi and Bacteria:
at 5,000 x g for 5 minutes. The supernatant was In the present investigation, a total of three
saved and the above procedure was repeated twice different genera of fungus were isolated and
to ensure complete extraction of the pigment. The identified. Among the genera Aspergillus with four
pooled supernatant was made upto a known volume
with 80 per cent methanol (to compensate the
solvent loss during heating). The absorbency was
measured at 663 nm in Spectronic 20 against
methanol blank.
Physico-chemical Analysis of distilleries effluent
The physico-chemical properties of the
distillery effluent like pH, Carbondioxide, alkalinity
(Carbonate and Bicarbonate), Dissolved oxygen,
Biological Oxygen Demand, Chemical Oxygen
Demand (COD), Nitrate, Nitrite, Ammonia, total
phosphorus, inorganic phosphate, Organic
phosphate, Calcium, Magnesium and Chloride were
analyzed following the procedure of Apha (1975).
Biochemical analysis of Nostoc muscorum
Biochemical analysis of Nostoc muscorum
such as Carbohydrate (Dubois et al., 1956), Protein
(Lowry et al., 1951), aminoacid. (Jayaraman, 1981)
and Lipids (Sato and Murata, 1988), were analyzed
Fig-1 Microbial diversity of distillery effluent
different cyanobacteria with different effluents has However, Nostoc muscorum was marginally better
already been reported by Vijayakumar et al. 2005 in removing all form of nitrogen. Suspended,
and Boominathan et al., 2007. This raise in DO cultivation of micro algae is one of the biological
level in effluents with cyanobacteria might be due process has been employed to eliminate residual
to oxygenic photosynthetic nature of cyanobacteria. inorganic nutrients as a tertiary treatment step from
The correlation between increase in DO and secondary treated effluents (Oswald, 1978). Studies
removal of BOD and COD observed in this study with Spirulina (Boominathan et al., 2007),
agree with observations of Kankal et al., (1987); Anabaena (Mallick and Rai, 1994), Oscillatoria
and Vijayakumar et al., (2005). (Manoharan and Subramanian, 1992 and 1993)
Suspended cultivation of microalgae is one concluded that cyanobacteria can efficiently
of the biological processes which have been eliminate inorganic nitrogen compounds from waste
employed to eliminate residual inorganic nutrients waters. In general, the removal of inorganic
as a tertiary treatment step from secondary treated nitrogen compounds by cyanbacteria is governed by
effluents (Prakasham and Ramakrishnan, 1998). growth conditions and physiological conditions of
Inorgornic compounds such as nitrite, nitrate, the state of the organism like pH, light, temperature,
ammonia and phosphate are the essential inoculum size and subtracted concentration.
requirements for the growth of cyanobacteria. They The capacity of cyanobacteria to remove
have high nutrient capabilities as they can large amount of phosphorus from industrial waste
accumulate inorganic phosphate and cynophycin water has been demonstrated by several workers
respectively (Fay, 1983). In the present (Monoharan and Subramanian (1992 and 1993),
investigation, in distilleries effluent, all form of Boominathan et al. (2007) and Vijayakumar (2005)
nitrogen were observed in appreciable quantities found a total or near total removal of all forms of
(Table-4) and cyanabacteria removed more than 55 phosphate by Oscillatoria and Aphanocapsa from
per cent of inorganic nitrogen from the effluent. different effluents. Tang et al., (1997) also reported
158 Journal of Research in Biology (2011) 3: 153-162
Ganapathy Selvam et al.,2011
a higher phosphate uptake, despite low biomass of treatment methods. However, in the present
cyanobacteria. Dash and Mishra (1999) observed investigation Nostoc was efficient by removing
100 per cent removal of phosphate from paper mill chloride from the sample (Table-4). Uma and
effluent while testing with Westiellopsis prolifica. Subramanian (1990) observed nearly 50 and 25 per
However in the present study more than 55 percent cent removal when ossein effluent which has very
removal of total phosphorous and 36 per cent high level of chloride was treated with Oscillatoria
removal inorganic phosphate were observed in and Aphanocapsa respectively. Manoharan and
effluent with Nostoc muscorum. The major Subramaniam (1992 and 1993) also observed more
phosphate reserve of cyanobacteria is than 40 percent removal of chloride from various
polyphosphate, which accumulates as discrete effluents by Oscillatoria sp. A similar observation
granules in the cytoplasm of the cell wall when attributing 50 percent chloride reduction under
phosphate is in excess (Fay, 1983). The intracellular laboratory conditions by O. brevis was also
accumulation of phosphate is energy dependent, reported in dye effluent (Vijayakumar et al., 2005),
being higher in the light than in the dark and oil refinery effluent (Boominathan et al., 2007) and
depends strictly on the pH of the cytoplasm. All the sugar mill effluent (Gopalakrishnan, 2007).
above findings confirm that cyanobacteria can Biochemical studies on Nostoc muscorum
absorb phosphate in excess amount as it is required Growth was measured in terms of
and this could be the reason for maximum removal chlorophyll a as a biomass component. This was
of all forms of phosphate from the effluent. estimated both in BG11 medium (control) and
The total hardness is constituted by calcium effluent. Maximum growth of Nostoc was recorded
and magnesium. In the present study more than 68 in the control than in effluent inoculated (Fig.2). In
per cent removal of calcium and magnesium were recent years, scientists have been increasingly
observed in effluent with Nostoc. Uma and concentrated more on the influence of these systems
Subramaniam (1990) studied the effective use of on the removal of nutrients from the effluent but
cyanobacteria in ossein effluent which has high only a few have investigated the effect of effluents
levels of calcium. They found more than 50 per cent on the biochemistry of the cyanobacterial systems.
reduction of calcium within 4 days when the To develop suitable and efficient treatment system,
effluent was treated separately with Oscillatoria it is obligatory to understand the mutual influence
and Aphanocapsa. Similarly, Manoharan and and interactions between the effluents and the
Subramanian (1992 and 1993) reported the organisms, so that manipulation to improve the
reduction of calcium and magnesium in domestic treatment system becomes feasible and hence the
sewage, ossein and paper mill effluents by O. present investigation on the biochemistry of
pseudogeminata var. unigranulata. They observed effluent grown cyanobacteria was carried out.
more than 70 percent reduction of calcium and Distilleries effluent has decreased the total
magnesium with retention time of 15 days. Dash carbohydrate content of Nostoc muscorum to a
and Mishra (1999) observed 50 per cent reduction considerable level (Fig.3) when compared to BG11
of calcium in paper mill effluent by Westiellopsis medium (control). The reduction was around 16
(retention time of 15 days). On the other hand percent. Contrary to the present observation,
Vijayakumar et al. (2005) reported more than 90 increase in the carbohydrate level of cyanobacteria
percent removal of calcium and magnesium from with various effluents has been reported by many
dye effluent when treated with Oscillatoria sp. workers (Reddy et al., 1983 and Vijayakumar,
Although, calcium is undoubtedly required for 2005, 2007). Nitrogen limitation caused photo
cyanobacterial growth substantial reduction in assimilated carbon to be directed towards the
calcium and magnesium are known to be essential synthesis of carbohydrate instead of proteins and
for flocculation and would coflocculate (Richmond chlorophyll. This response is widely observed in
and Becker, 1986).) observed the excretion of many algal species (Turpin, 1991). Protein and
organic acids by microbes and especially chlorophyll decreases and carbohydrate increases
cyanobacteria and their capacity to solubilize by CO2 enrichment have been observed previously
magnesium in the waste water, which could explain in a number of species (Loehle, 1995). But in the
observed reduction. present investigation, considerable levels of all
Chlorides are generally considered as one of forms of inorganic nitrogen were observed; more
the major pollutants in the effluent which are over the carbohydrate content was also reduced by
difficult to be removed by conventional biological the effluent. There were no CO2 recorded in the
Journal of Research in Biology (2011) 3: 153-162 159
Ganapathy Selvam et al.,2011
effluent throughout the study period and hence this Bakare AA, Lateef A, Amuda OS and Afolabi
could not explain the observed variation in RO. 2003. The aquatic toxicity in characterization
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Authors: ABSTRACT:
Ganesan CM and
Paulsamy S.
Email:
Keywords:
paulsami@yahoo.com In vitro culture, Medicinal plant, Artemisia annua, Nilgiris, Western Ghats,
bioganesan@gmail.com. India.
Dates:
Received: 04 Jul 2011 /Accepted: 08 Jul 2011 /Published: 12 Jul 2011
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.
factors (Ananthi et al., 2011). The amount of leaf endogenous level of growth regulators as observed
explant responding for callus formation was in many other plants (Farternale et al., 2002;
ranging between 10 and 84% (Table 1). MS Senthilkumar and Paulsamy, 2010b). It was noted
medium fortified with NAA at 0.9 mg/l initiated that the NAA alone or in combination with TDZ
98.66% of leaf discs for callus formation (Fig. 1a) generally have the efficiency of initiation at high
followed by 0.7mg/l of NAA initiated 84% of leaf percentage of (>50%) leaf explant for callus
discs for callusing and 0.5mg/l of NAA and TDZ formation. It indicates the higher requirement of
each initiated 73% of discs for callusing. The other certain auxins like NAA alone or in combination
combinations and concentrations of growth with low quantity of cytokinins like TDZ for callus
hormones in the medium initiated around only 20 to formation of the study species, A. annua.
50% of leaf discs of A.annua for callus formation. Karappusamy and Pullaiah (2007) for the species,
Baskaran and Jayabalan (2005) explained that the Bupleurum distichophyllum and Senthilkumar and
differential response of same or different explants Paulsamy (2010a) for the species, Ageratum
for callus formation could be due to the nature of conyzoides also have reported effective callus
tissue, degree of totipotency and composition of formation from the leaf explants in the medium
medium with respect to micronutrients and containing high quantity of NAA. Mariani et al.,
hormones. Further it is explained that the variation (2011) reported the requirement of the cytokinin
in response of discs in terms of callus initiation may like compounds, TDZ for effective callus formation
be due to the variation in distribution of in the ornamental plant, Aglaonema sp. The colour
Table 1. Effect of different concentrations of growth regulators on per cent callus induction from leaf, node
and intermodal explants of the species, Artemisia annua.
Days required for
Growth regulator Callus formation Colour of the
callus formation after
(mg/l) (%) callus
inoculation
BAP NAA 2,4-D IBA TDZ Leaf explant Leaf explant Leaf explant
0.5 0 0 0 0 10 10.002.00a G
1.0 0 0 0 0 12 13.661.52a G
1.5 0 0 0 0 14 18.002.00b G
2.5 0 0 0 0 16 21.331.53bc G
3.0 0 0 0 0 18 23.331.58c G
0 0.1 0 0 0 18 25.002.00c G
0 0.3 0 0.5 0 21 29.002.00cd G
0 0.5 0 1.0 0 20 30.001.00d DG
0 0.7 0 1.5 0 23 49.002.00d DG
0 1.0 0 2.0 0 22 57.332.51e DG
0.5 0 0.5 0 0 18 39.001.00f G
1.0 0 1.0 0 0 17 44.332.08d DG
1.5 0 1.5 0 0 15 46.661.52d DG
2.0 0 2.5 0 0 21 49.332.08d LG
2.5 0 3.0 0 0 19 51.332.08d LG
3.0 0 3.5 0 0 23 55.001.00e B
0 0.1 0 0 0 14 39.661.53f LG
0 0.3 0 0 0 18 49.661.55d LG
0 0.5 0 0 0 20 46.661.59d B
0 0.7 0 0 0 21 84.004.35g LB
0 0.9 0 0 0 30 98.662.51h LB
0 0.5 0 0 0.1 22 54.331.58e DB
0 0.5 0 0 0.2 19 54.660.57e DB
0 0.5 0 0 0.3 17 56.662.08e DB
0 0.5 0 0 0.4 25 58.003.00e DB
0 0.5 0 0 0.5 23 73.004.00i LB
G-Green, DG- Dark green, LG- Light green, B-Brown, DB-Dark brown, LB- Light brown
Means in column followed by different letter(s) are significantly different at 5% level according to DMRT.
of the calli was showing wide degree like green, species (Vijaykumari et al., 2001; Roy et al., 2008;
dark green, light green, brown, dark brown and Senthilkumr and Paulsamy, 2010a; Sunder and
light brown according to the combinations and Jawahar, 2011).
concentrations of the growth regulators in the MS The rooting attributes of A.annua while
medium (Table 1). subculturing the secondary explant shoots were
The results of the subculturing experiments well pronounced in the MS medium supplemented
by using the secondary explant, leaf derived callus with the auxin, IBA alone at higher concentrations
showed that the cytokinin, BAP alone in higher from 0.5 to 0.9 mg/l (Table 3). The IBA
concentration (>1.5mg/l) (Fig. 1b) or BAP in concentration at 0.9 mg/l initiated 85% shoots for
combination with GA3 have enhanced the response root formation (Fig. 1d) followed by 0.7mg/l
of calli for shoot formation by 98 and 90% initiated 80% and 0.5 mg/l initiated 71% shoots for
respectively (Table 2). In addition, greater number root formation. The number of roots per shoot were
of 20 shoots/callus was also noted to be produced also observed to be higher (12 roots/shoot) in the
while subculturing the calli on MS medium with MS medium containing 0.9mg/l IBA for the study
BAP and GA3 at 2.7 and 0.9 mg/l respectively species, A.annua. Similarly, the root length was
(Table 2) (Fig. 1c). However, the higher shoot greater (5.1cm) during the subculturing of in vitro
length of 10cm was achieved in the MS medium cultured shoots for roots on MS medium with IBA
fortified with BAP alone at 2 mg/l (Table 2). All at 0.9 mg/l. All these facts showed that the auxin,
these facts indicate that the cytokinin, BAP is the IBA is the most required growth regulator for
most essential growth regulator for effective shooting characters of the study species, A.annua.
shooting of the study species, A.annua. It is of It agrees with the concept that auxins are the plant
common fact that cytokinin is the major growth hormones endogenously or exogenously inducing
hormone involved in shoot formation in many plant root formation in majority of plant species (Van
Table 2. Effect of different concentrations of growth regulators on shoot initiation, shoot number and shoot
length after subculturing the leaf derived callus of the species, Artemisia annua.
Growth regulator (mg/l)
Culture response (%) No. of shoots/callus Shoot length (cm)
BAP IAA GA3
0.5 0.0 0.0 57.332.08ag 08.661.52a 07.570.35a
b a
1.0 0.0 0.0 65.663.51 09.002.00 07.930.57a
c b
1.5 0.0 0.0 85.333.05 12.001.00 08.830.74a
d b
2.0 0.0 0.0 98.002.00 14.001.00 10.200.26b
0.5 0.2 0.0 20.333.05e 06.331.53a 04.800.30c
e a
1.0 0.4 0.0 29.661.53 07.331.53 06.500.20ca
af a
1.5 0.6 0.0 48.332.08 09.671.54 07.000.20a
b b
2.0 0.8 0.0 62.334.51 14.002.00 07.370.15a
d cd
3.0 1.0 0.0 93.002.00 17.661.56 08.500.36a
f b
0.3 0.0 0.1 45.002.00 12.331.53 05.030.25c
0.6 0.0 0.2 50.333.21f 15.002.00bd 05.130.15c
a bd
0.9 0.0 0.3 54.003.00 15.671.53 05.230.15c
a bd
1.2 0.0 0.4 55.332.08 16.671.53 05.200.10c
a cd
1.5 0.0 0.5 55.331.53 17.002.00 05.300.10c
g cd
1.8 0.0 0.6 59.673.05 18.002.00 05.670.15c
2.1 0.0 0.7 70.331.53b 19.671.57cd 05.900.20c
c cd
2.4 0.0 0.8 80.001.00 19.002.00 05.900.10c
e cd
2.7 0.0 0.9 90.002.00 20.332.08 06.670.15ca
e cd
0.5 0.0 1.0 94.672.08 20.671.58 07.100.20a
af a
1.0 0.2 1.0 46.332.52 07.002.00 02.870.20d
1.5 0.4 1.0 50.672.52f 06.671.53a 02.930.15d
f a
2.0 0.6 1.0 51.332.52 06.001.00 03.000.10dc
g b
2.5 0.8 1.0 59.332.08 07.001.00 03.470.25dc
b a
3.0 1.0 1.0 74.002.00 11.331.53 04.700.20c
Means in columns followed by different letter(s) are significantly different at 5% level according to DMRT.
176 Journal of Research in Biology (2011) 3: 173-178
Ganesan et al.,2011
Table 3. Effect of different concentrations of growth regulators on root number, rooting percentage and
root length after subculturing the leaf calli derived shoots of the species, Artemisia annua.
Growth regulator (mg/l)
Shoots rooted (%) No. of roots/shoot Root length (cm)
IBA IAA NAA
0.1 0.0 0.1 19.001.00a 05.000.81a 2.100.20a
a b
0.2 0.0 0.2 20.661.52 07.000.82 2.700.10ab
0.3 0.0 0.3 22.672.51a 08.000.85b 3.030.15b
b c
0.4 0.0 0.4 27.331.57 10.661.24 3.730.16b
0.5 0.0 0.5 29.001.00b 08.661.26b 2.801.90ab
a b
0.0 0.1 0.1 22.331.52 07.331.27 3.460.15b
0.0 0.2 0.3 26.001.00b 10.002.44c 3.530.15b
0.0 0.3 0.5 29.671.54b 09.331.21c 3.630.15b
ce b
0.0 0.4 0.7 40.001.00 07.001.64 3.800.10b
0.0 0.5 0.9 46.001.01d 09.001.63c 4.060.17b
ce cd
0.0 0.1 0.0 39.001.05 10.671.28 2.170.21a
0.0 0.2 0.0 41.672.08ce 08.671.23b 2.700.20ab
ce c
0.0 0.3 0.0 42.671.54 10.001.63 3.000.13b
0.0 0.4 0.0 44.681.52df 09.002.16c 2.800.10ab
f b
0.0 0.5 0.0 48.331.27 08.671.25 3.130.25b
0.1 0.0 0.0 50.662.51f 07.001.63b 3.200.27b
g c
0.3 0.0 0.0 60.001.00 09.661.26 4.030.15bd
0.5 0.0 0.0 71.002.00h 11.661.27cd 4.430.31bd
0.7 0.0 0.0 80.001.00i 10.331.20cd 4.800.20d
j d
0.9 0.0 0.0 85.001.00 12.000.81 5.100.10d
1.0 0.0 0.1 41.661.52ce 11.331.27cd 3.260.15b
f c
1.0 0.0 0.2 47.002.00 09.001.63 3.600.20b
1.0 0.0 0.3 50.662.08f 07.331.25b 3.730.20b
g a
1.0 0.0 0.4 56.001.00 05.000.81 3.730.15b
1.0 0.0 0.5 60.001.00g 06.000.83a 3.900.10b
Means in columns followed by different letter(s) are significantly different at 5% level according to DMRT.
Eck and Kitto, 1992). Similar kind of findings of The present paper describes a prime and
effective root formation by the influence of various easy-to-use protocol for large scale production of
types of auxins in many plant species have been plantlets of A.annua through leaf culture and the
reported elsewhere (Mallikadevi and Paulsamy, method is useful for the ex situ conservation of this
2009; Mahesh et al., 2010; Loc et al., 2011; species as well. In addition, the findings of the
Mungole et al., 2011; Rajput et al., 2011). present investigation provide a baseline data for
The hardening experiments showed that high further research in this species.
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Authors: ABSTRACT:
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MohanPM, DhivyaP,
Muruganandam2N,
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DNA barcode technique has been recently promoted as a method for both
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Islands, India.
Corresponding author:
Keywords:
Dr. PM. Mohan, Ph.D., DNA barcode, Serranidae, Phylogenetic and Andaman Sea.
Dates:
Web Address: Received: 04 Jul 2011 /Accepted: 08 Jul 2011 /Published: 18 Jul 2011
http://jresearchbiology.com/
Documents/RA0056.pdf.
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.
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Fig. 1. Neighbor-joining trees based on the mtDNA COI gene nucleotide sequences of Plectropomus species
analyzed. Numbers at nodes are bootstrap values based on 1000 replicates. The scale bar represents an interval
of Tamura-Nei genetic distance for Plectropomus leopardus.
65
EF607554.1 Seriola dumerili
99 FJ237809.1 Lutjanus lutjanus
EU600141.1 Epinephelus fario
99 FJ237765.1 Epinephelus bleekeri
33 EU600147.1 Epinephelus akaara
HQ174862.1 Epinephelus fuscoguttatus
FJ583012.1 Cephalopholis urodeta
DQ107868.1 Epinephelus areolatus
81
DQ107886.1 Epinephelus multinotatus
36
FJ237772.1 Epinephelus spilotoceps
33 6 EF609352.1 Epinephelus undulosus
EF60922.1 Epinephelus longispinis
18 FJ237734.1 Epinephelus amblycephalus
FJ583397.1 Epinephelus ongus
99 EU266383.1 Epinephelus moara
7 FJ594964.1 Epinephelus bruneus
3
GU673644.1 Perciformes sp.
EF609518.1 Epinephelus diacanthus
4
FJ583396.1 Epinephelus adscensionis
3
EU595118.1 Epinephelus sexfasciatus
The comparative analysis of the Optical Density for MRET activated water and non-
activated water in visible, ultraviolet, and infrared regions of the spectrum
Journal of Research in Biology
Authors: ABSTRACT:
Igor Smirnov MRET activated water is produced with the help of the USA patented
Molecular Resonance Effect Technology (MRET). The anomalous electrodynamic
characteristics (dielectric permittivity and electrical conductivity) of MRET water
subjected to applied electromagnetic field in the area of very low range of frequencies
and the anomalous viscosity of MRET water subjected to very low tangential pressure
Institution:
Affiliation: Global have provided some evidence for the long range dynamic polarized-oriented
Quantech, Inc. Encinitas, multilayer structuring of such activated water .
California, USA. The objective of this article is to demonstrate the relatively high, long range
dynamic structuring of water molecules in activated water produced with the help of
the MRET activation process. To achieve this goal there were conducted a number of
the comparative analysis of the optical density for MRET activated water and non-
activated water in visible, ultraviolet (UV), and infrared (IR) regions of the spectrum.
Corresponding author: This analysis confirms that MRET activation process has the tendency to force water
Igor Smirnov molecules to form large size clusters in the volume of water.
Such findings provide some evidence that MRET activation leads to the
formation of polarized-oriented multilayer structuring of water molecules. The
correlation analysis of the scattered laser emission also provides the unambiguous
confirmation of the presence of the stable water memory phenomenon. The duration
Email: of water memory phenomenon was observed for many hours and days in this
igor@gqusa.com
particular experiment. The water memory phenomenon can be interpreted as the
existence of stable super-molecular clusters in the volume of activated water.
Article Citation:
Igor Smirnov.
The comparative analysis of the Optical Density for MRET activated water and non-
activated water in visible, ultraviolet, and infrared regions of the spectrum.
Journal of research in Biology (2011) 3: 184-190
Dates:
Received: 21 Jun 2011 /Accepted: 24 Jun 2011 /Published: 18 Jul 2011
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.
Fig. 1 Optical density of non-activated water (1) and Fig. 2 IR-spectra of non-activated water (upper
MRET water activated for 30 min (2) versus the curve) and activated water that was studied in 5
wavelength in the visible and UV ranges. Both plots hours after the activation (middle curve) and in 1
visually coincide with each other [Vysotskii et al., 2009]. hour (lower curve) [Vysotskii et al., 2009].
studied by the method of infrared Fourier- conducted on four different samples of the initial
spectroscopy with the help of a device Nexus non-activated water (see Fig.5):
produced by the firm Thermo Nicolet. The 1. Natural Mineral Water AVIAN (produced in
studies were performed in the range of wave France);
numbers from 50 to 4000 cm1 (Fig.3). 2. FreshWater Life JINBO/Seok Su (produced
MRET activated water was also studied by in the Republic of Korea);
Raman scattering spectroscopy. The Raman 3. Natural Mineral Water HAITAI/Gangwondo/
scattering spectra for non-activated water and for Pyeong Chang (produced in the Republic of
MRET activated water was studied in 24 hours after Korea);
the completion of the activation and are presented 4. Natural Mineral Water JEJU/Sa Da
in Fig. 4. The studies were performed with the same Soo (produced in Iceland).
fraction of water (activation for 30 min). The
spectra determine the Raman scattering RESULTS:
characteristics in the region from 1 cm1 to 4000 Visually, both plots in Fig.1 coincide with
cm1. each other. This corresponds to that almost no
With the purpose to discover the presence of difference between the spectra of the initial non-
structural elements in the water bulk there carried activated water and activated one observed in the
out an additional study in the optical characteristics visible and UV regions of the absorption spectrum
of MRET activated water with the help of the laser in the limits of accuracy of the method (0.5%),
correlation analyzer AKVA-01. The principle of except for a small increase in the absorption, at
action of this device is based on the analysis of the most 12%, in the UV region of the spectrum at
characteristics of a mutual temporal coherence wavelengths near = 190210 nm.
function of the laser emission scattered in the This result differs basically from the above
volume of the studied water in the direction mentioned very essential changes in the
perpendicular to the initial laser beam. There was electrodynamics characteristics of activated water
used the emission with a wavelength of = 0.63m in the region of low frequencies. It shows that the
generated by a HeNe laser was used in the process of activation by means of MRET activation
analysis. The longitudinal coherence length of such does not lead to a significant change of the electron
an emission prior to its scattering is very large and subsystem of atoms and involves only structural
exceeds hundreds of meters. The width of the and juxtaposition changes of the water molecular
spectrum of this emission is very small [Vysotskii system.
et al., 2009]. The results of the study in various regions of
The laser spectroscopy measurements were IR-spectra are presented in Figs. 2. This diagram
Fig. 3 IR-spectrum of non-activated water (upper Fig. 4 Raman scattering spectra of non-activated (1)
curve) and water activated for 30 min (lower curve). and activated (2)water. Fragments of the spectra (1a)
The spectrum was studied in 1 hour after the and (2a) correspond to the increase in the scale by 10
activation [Vysotskii et al., 2009]. times. All spectra were measured in 24 hours after
the activation [Vysotskii et al., 2009].
gives the IR-spectra in the range of 50 550 cm1
for the initial non-activated water and MRET water cm1. This result is presented in Fig. 3 for non-
activated for 30 min. The study was carried out activated water and water activated for 30 min (the
with three samples of water derived from the spectrum of activated water was studied in 1 hour
identical distilled water: after the activation).
Initial distilled water; MRET activated water was also studied by
MRET activated water stored after the Raman scattering spectroscopy. It is seen from the
activation prior to the measurement for 30 min data presented in Fig. 4 that MRET activation of
at room temperature; water, it leads to a small increase of the Raman
MRET activated water which was stored after scattering amplitude in the whole range from 1 cm1
the activation prior to the measurement for 5 to 4000 cm1.
hours at room temperature. The optical characteristics of MRET
Such study allows us to determine the activated water was additionally studied with the
dependence of the influence of the activation on the help of laser spectroscopy.
IR-spectrum of water, as well as the influence of the As a quantitative characteristic there was used the
storage duration of this water on its properties. It is integral index of the molecular dynamics of water
seen from Fig. 2 that a decrease in the absorption W which is inversely proportional to the coefficient
of water (a decrease of the imaginary part of the of diffusion D of the scattering objects including
dielectric permittivity) in the long wave part of the the normalizing factor (in order to make W
IR- occurs in the process of MRET activation. This dimensionless).
decrease exceeds 10% for the range of wave The parameters of the scattered emission
numbers less than 300 cm1. depend on the motion of scattering molecules of
With increase of the wave number, the water and the ensembles of these molecules. Due to
absolute value of a change in the coefficient of the presence of the fluctuation (diffusive)
absorption remains approximately constant, and the mechanical, motion of the scattering objects, the
relative change decreases, respectively. With phase of the scattered emission continuously
increase of the duration of storage of activated fluctuates, which decreases very sharply the
water, the discovered change in the optical density coherence length of the scattered laser field. A
in the IR-range gradually decreases. change in the coherence length of the emission and
The effect of a small decrease in the optical the associated change in the spectrum of the
density of MRET activated water is also observed emission are those parameters of which the
in the region of great wave numbers 10004000 measurement allows us to determine the
187 Journal of Research in Biology (2011) 3: 184-190
Smirnov et al.,2011
DISCUSSION:
The obtained experimental data provides
evidence that the specific features of the spectrum
of MRET activated water in IR, visible, and UV
ranges preserve for a long time after the completion
of the activation, though the very changes in the
spectrum in these ranges turned out to be a small
Fig. 5 Integral index W of the molecular dynamics ones. Such changes in the optical properties of
of activated water of different types versus time. The MRET activated water which are small in
numbers stand for the types of water mentioned magnitude but with the long time of their
above in the text [Vysotskii et al., 2009].
preservation can be satisfactorily substantiated if we
characteristics of the motion of scattering objects consider that they are related to a change in the
(the coefficient of diffusion) and, hence, the mass properties of a relatively small number of separate
and size of these objects. The coherence length can molecules of water isolated from the external action
be found with the help of the autocorrelation and corresponding to the hindered relaxation. Based
function (1.1). on the proposed model of the memory of water, we
It is seen (Fig.5) that the index W was can refer such changes to the molecules of water
changed comparatively slightly (except for water being in the volume of clathrate microcavities
sample #3) for the time of the first activation [Vysotskii at el., 2005].
process. Thereafter for 24 hours (with an interval of It was proposed earlier that there exists a
two hours) there was measured the index W for all strong repulsive electrostatic field in such
types of water. It is seen that, for this time interval, microcavities. For this reason, the internal walls
there occurred a systematic increase of the index W of microcavities possess the hydrophobic properties
for all types of water except for water #3. In this and do not form hydrogen bonds with molecules of
case, a change in W was accompanied by very water placed in the volume of microcavities. These
strong oscillations for the water samples #2 and #3. molecules, due to the absence of a hydrogen bond
At the same time, the index W for all the remaining with molecules of water forming the walls of
types of water was changed as a monotonously microcavities, have a changed configuration of the
increasing function of time. There was performed electron shell (it corresponds to a free H2O
the second activation of water in 24 hours after the molecule, rather than that of a bound molecule) and,
first activation with the same duration of one hour. respectively, some different optical properties
The results of this activation also turned out [Vysotskii at el., 2009].
ambiguous. After the activation, there was observed The process of MRET activation renders a
a very sharp increase of the index W for water strong influence on the dynamics of the mechanical
sample #4. At the same time, this index was fluctuation motion of stable global structural
practically constant for the remaining samples of elements in the water bulk. The formation and the
water. The measurements after the second long-term existence of such structural elements
activation were performed for 42 hours (At first, depend on the activation of water and confirm the
three measurements were carried out with an possibility for the memory of water to exist. To find
interval of two hours and then the other three such structural formation in the body of MRET
measurements with an interval of 12 hours). After activated water there was used the laser
the completion of the whole cycle of measurements, spectroscopy method. The study of the correlation
the values of the index W for all samples of water spectrum allows us to determine the mean
(except for water sample #3) turned out to be characteristics of the motion of scattering objects in
exceeding the initial value W0 (prior to the first the water bulk and, on this basis, to perform the
qualitative analysis of the spatial structure of water. size of a stiffly bound molecular complex which is
In addition, such studies allow one to determine the present in the water bulk and scatters the laser
dependence of these characteristics on the emission. There was performed the following
activation of water. procedure and the sequence of studies that total
In order to determine the regularities of the duration was equal to 71 hours. First of all there
process of scattering, it is necessary to determine was carried out three measurements of the index W
those quantities which are not random and describe for 3 hours for each of the studied types of water.
unambiguously the scattering. The deterministic The purpose of these measurements were related to
(nonrandom) characteristics of the scattered the determination of the stability of a measuring
emission can be found if the autocorrelation unit. The results of this testing period correspond
function of field is known: to the first three points on all the plots presented in
Fig. 5. The very small variance of the index W
confirms a high degree of stability, a small value of
the apparatus error, and the error related to the
procedure of measurements. It is obvious that the
differences of the indexes of molecular dynamics W
for different types of water prior to the activation
are related to a great extent to the differences of
(1.1) their salt composition. This is explained by the fact
Where: E0 is the amplitude of an incident wave; that the ions dissolved in water influence the mass
and size of molecular complexes in water and,
is the scattering amplitude of the
hence, the coefficient of diffusion of these
emission;
complexes and the spectrum of the scattered laser
emission. It is possible to assume that such an
is the wave vector scattered by an angle ; influence can be realized in at least two ways: on
R is the distance from the region center in the water the one hand, the dissolved ions break the chemical
bulk, where the objects scattering the light are homogeneity of water and therefore affect the
positioned; formation of molecular complexes formed from
N is a number of scattering objects in the volume molecules of water; on the other hand, the union of
of water; dissolved ions with these complexes changes the
E is the amplitude of the scattered electric field density and the mass of the latter. It is obvious that
intensity; both mechanisms lead to the essential influence of
D is the diffusion coefficient of scattering objects; dissolved salts on the spectrum of the scattered
The spectrum of the scattered emission is emission, which was observed in the study of
calculated with the help of the Wiener Khinchin various samples of water.
formula: After the completion of the whole cycle of
measurements, the values of the index W for all
samples of water (except for water sample #3)
turned out to be significantly exceeding the initial
value W0 (prior to the first activation) that allows to
assume the formation of the water molecular super-
clusters in the volume of water after MRET
activation process.
(1.2) The results of the optical study of MRET
It is seen from formula (1.2) that the activated water in UV, IR spectrum, and Raman
spectrum of the scattered emission depends on the scattering spectra also provide evidence of the
coefficient of diffusion D defining the mean stable polarized-oriented multilayer molecular
characteristics of the motion of scattering objects. formation in activated water. It shows that the
This coefficient, in turn, depends on the size and process of activation does not lead to a significant
mass of these objects. change of the electron subsystem of atoms and
An increase of the index W corresponds to a involves only structural and juxtaposition changes
decrease of the coefficient of diffusion and of the water molecular system. The confirmation of
characterizes uniquely the increase of the mass and such molecular formations can explain the
189 Journal of Research in Biology (2011) 3: 184-190
Smirnov et al.,2011
ACKNOWLEDGEMENT:
I deeply acknowledge Prof. Vysotskii V. I.
and Kornilova A.A., Ph.D. Moscow State
University, Russia, and Helios Alpha (USA) for
their help in my research work.
Authors: ABSTRACT:
Upadhyaya S1 and
Saikia2 PK. Morphomatric of Cotton Pygmy-goose, Nettapus coromandelianus
coromandelianus was studied during 2006 to 2008. The males are comparatively
bigger in size than the females. The average weight was found to be 226.50 gm and
219.50 gm for male and female respectively. The primary (wing) feather arrangement
Institution:
1. Deptt. of Zoology, in male was found to be P1 < P11 < P10 < P9 < P8 < P7 < P2 < P6 <P5 < P3< P4. The
Tyagbir Hem Baruah females have a more or less similar arrangement except the P2 and P6 where P6<P2.
College, Karchantola, The mean length of the middle toe in male was found to be 34.32+0.194 mm; where
Dist.-Sonitpur (Assam), as in female the same remains 0.5 mm shorter (+0.163). The wing expansion was
India. PIN-784189. ranged between 424 mm to 426 mm in both male and female, but with slight variation
2. Department of Zoology, in mean value (male- 425.17+0.753 mm; female- 425.53+0.816 mm). Since no
Gauhati University, Assam morphomatric studies has been done so far on this species, the present paper was
India. hypothesized to the study of morphomatric variation in various aspects of Cotton
Pygmy-goose indicating the relation of wings, hind-limbs, head neck, beak, tarsus,
different types of toes and tail in respect to the habitat utilization and ecology of the
wetland.
Corresponding author:
Dr. Sanjib Upadhyaya
Email: Keywords:
sanjib1970@sify.com Eco-morphology, middle toe, tarsus and wing expansion.
Article Citation:
Web Address: Upadhyaya S and Saikia PK.
http://jresearchbiology.com/
Documents/RA0051.pdf.
Study of morphomatric biology of cotton Pygmy-Goose Nettapus coromandelianus
coromandelianus Gmelin.
Journal of research in Biology (2011) 3: 191-201
Dates:
Received: 27 Jun 2011 /Accepted: 08 Jul 2011 /Published: 18 Jul 2011
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.
same found to be 227.0 gm + 6.8SD (range= 215.5 =332.9 to 336.5 mm, n =27), whereas the mean
to 236.34 gm, n =15). A greater BoW (in either sex) BoL (BT to TT) measured in female was 351.5 +1.6
was found during the monsoon season with a mean mm (range =350.6 to 353.4 mm, n =15). Seasonal
of 225.6 gm +20.4SD (male: mean 222.1 gm variation was observed in BoL (BT to TT) of goose
+22.2SD, range =180.65 to 260.0 gm, n =13; during the study period. In winter season, a greater
female: mean 229.08 gm +18.6SD, range =221.7 to mean BoL (BT to TT) was observed in female with
234.5 gm, n =6). 351.2 mm +1.5SD (range =348.8 to 353.2 mm, n
b. Body length (BoL) =9) than the male with a mean BoL (BT to TT) of
The mean BoL (BT to TT) of Cotton Pygmy 334.8 mm +1.2SD (range =332.7 to 336.1 mm, n
-goose, irrespective of sex was 335.3 mm + 1.1SD =14), while the case is reversed during monsoon
(range =332.9 to 353.4 mm, n =42, Table 5.1, (Female, mean: 351.8 mm +1.7SD, range: 349.1 to
Figure 5.1). In male goose, the mean BoL (BT to 353.4 mm, n =6; Male, mean: 335.8 mm +0.7SD,
TT) measured was 335.3 mm +1.1SD (range range: 333.9 to 336.5 mm, n =13). The mean BoL
(N to A) of the goose was found to be 154.7 mm +
1.1SD (range =143.5 to 165.0 mm, n =42, Table
5.1, Figure 5.1) irrespective of sex and seasons. The
mean BoL (N to A) was found higher in female
with a mean length 163.6 mm +1.2SD (range
=162.0 to 165.0 mm; n =15) than the male which
has a mean BoL (N to A) of 145.7 mm + 1.1SD, n
=27) irrespective of the seasons (Table 5.1). The
male and females were found with a greater BoL (N
to A) during monsoon then the winter (male
(a) (b) monsoon: mean BoL 146.0 mm +1.1SD, range
Plate 1 Primary wing feathers of (a) male & (b) =143.3 to 147.0 mm, n =13; winter: mean BoL
female CPG (from inner to outer)
145.4 mm +0.9SD, n =14; female winter: 163.5 mm female (male: mean 15.4 mm+0.2SD, n =13;
+1.3SD, n =9 & monsoon: 163.9 mm +1.1SD, female: mean 15.3mm +0.1SD, n =21) during the
range =162.7 to 165.0 mm, n =6). monsoon season of a year.
c. Bill length (BL) e. Bill width (BW)
The mean BL (BT to F) of the Cotton The overall mean BW (at culmen) in Cotton
Pygmy-goose was found to be 26.5 mm +0.2SD Pygmy-goose was found to be 12.03 mm +0.2SD
(range =26.2 to 26.9 mm, n =42, Table 5.1, Figure (range 11.5 to 12. 7 mm, n = 42, Table 5.1, Figure
5.1) and BL (BT to F) between 22.3 mm to 23.2 5.1). The BW of the male goose was found to be
mm (mean 22.7 mm +0.3SD, n =12) with p >0.05 12.3 mm +0.2SD (range =12.0 to 12.7 mm, n =27),
(Students t-test) in both the cases irrespective of whereas in females the same found to be 11.8 mm
sex and seasons of a year. The male goose was of +0.2SD (range =11.5 to 12.0 mm, n =15). The BW
greater BL (BT to F) than the females with mean (at culmen) in case of male goose during monsoon
length 26.6 mm +0.2SD (range 26.2 to 26.9 mm, n was found to be 12.3 mm +0.2SD (range =12.0 to
=27) and 26.5 mm +0.2SD (range 26.2 to 26.8 mm, 12.5 mm; n =13), whereas the mean BW remains
n =15) respectively. The BL (BT to F) of the goose at12.4 mm +0.3SD (range =12.0 to 12.6 mm; n
was found longer during monsoon with a mean =14). Again the mean BD of female goose during
length of 26.6 mm +0.2SD (range =26.2 to 26.9 monsoon was found to be 11.4 mm +0.2SD (range
mm, n=19) than the winter period with a mean of =11.4 to 11.9 mm, n =6) and during winter it
26.5 mm +0.2SD (range =26.2 to 26.9 mm, n =23). remains at 11.8 mm +0.2SD (range =11.5 to 12.0
No major differences were observed in the BL (BT mm, n =9).
to F) between the male and female during winter f. Head circumference (HC)
season (male: mean =26.5 mm +0.2SD, range =26.2 The mean HC of Cotton Pygmy-goose was found
to 26.9 mm, n =14; female: mean =26.5 mm to be 93.5 mm +0.1SD (range =92.7 to 94.2 mm, n
+0.2SD, range =26.2 to 26.8 mm, n =9). The mean =42, Table 5.1, Figure 5.1). The mean HC of male
BL (BT to FC) of the goose was found to be 22.8 goose was 93.9 mm +0.2SD (range =93.6 to 94.2
mm +0.2SD (range =22.3 to 23.2 mm, n =42). The mm, n =27) and of female goose was 92.9 mm
BL (BT to FC) in case of the male goose was found +0.2SD (range =92.7 to 93.2 mm, n =15)
greater with a mean 22.8 mm +0.3SD (range =22.4 irrespective of seasons of a year. The HC was found
to 23.2 mm; n =27), than the females where the greater during the monsoon season (93.5 mm
mean BL (BT to FC) was 22.6 mm +0.2SD (range +0.2SD, n =19) than during the winter season (93.4
=22.3 to 22.9 mm, n =15, Table 5.1). The Cotton mm +0.2SD, n =23) irrespective of the sex. During
Pygmy-goose, during the monsoon were found with the monsoon the male goose has a greater HC with
greater BL (BT to FC) than the goose observed a mean of 93.9 mm +0.2SD (range =93.6 to 94.2
during the winter (monsoon: mean 22.8 mm mm, n=13) than the female goose of the same
+0.9SD, range =22.4 to 23.2 mm, n =19; winter: season which has a mean HC of 93.0 mm +0.2SD
mean 22.7 mm +0.3SD, range =22.3 to 23.2 mm, n (range =92.7 to 93.2 mm, n =6).
=23). Again, the monsoon male were found to have g. Neck length (NL)
greater BL (BT to FC) than the monsoon females The mean NL of the Cotton Pygmy-goose was
(male: range 22.4 to 23.2 mm, mean 22.9 mm found to be 42.1 mm +0.6SD (range =40.0 to 43.5
+0.4SD; female: range 22.3 to 22.9 mm, mean 22.6 mm, n =42, Table 5.1, Figure 5.1) irrespective of
mm +0.2SD). sex and seasons of a year. The mean NL measured
d. Bill depth (BD) in male Cotton Pygmy-goose was 42.6 mm +0.4SD
In Cotton Pygmy-goose, the mean BD was (range= 42.0 to 43.5 mm, n =27), whereas in female
found to be 15.4 mm +0.1SD (range =15.1 to 15.6 goose the mean NL found was 41.4 mm +0.7SD
mm, n =42, Table 5.1, Figure 5.1). The male goose (range =40.0 to 42.1 mm, n =15. During the winter
has longer BD with a mean 15.4 mm+0.2SD (range season, the mean NL of the male was found to be
=15.1 to 15.6 mm, n =27), than the females who has 42.6 mm +0.4SD (range =42.0 to 43.2 mm, n =14),
a mean BD of 15.3 mm +0.1SD (range =15.1 to whereas the female has a mean NL of 41.3 mm
15.4 mm, n =15). During winter, the male goose has +0.7SD (range =40.0 to 42.1 mm, n =9). The
a mean BD of 15.4 mm +0.2SD (range =15.1 to female has mean NL of 41.8 mm +0.7SD (range
15.6 mm; n =14), whereas the female has a mean =40.5 to 42.1 mm, n =6), the male goose during
BD of 15.3 mm +0.1SD (range: 15.1 to 15.4 mm; n monsoon has a mean NL of 42.7 mm +0.5SD
=9). The BD found slightly greater in male than the (range = 42.0 to 43.5 mm, n =13).
194 Journal of Research in Biology (2011) 3: 191-201
Upadhyaya et al.,2011
h. Neck circumference (NC) (PWF) and 14-secondaries (SWF). The mean PWF
The mean NC of Cotton Pygmy-goose was found length was found to be 104.2 mm +0.06SD (range
to be 86.9 mm +0.2SD (range =86.5 to 87.2 mm, n =101.13 to 107.31 mm, n =42) irrespective of sexes
=42, Table 5.1, Figure 5.1) irrespective of sex and and seasons. The male goose has a mean PWF of
seasons of a year. The NC was slightly of greater 107.2 mm +0.07SD (range =107.07 to 107.31 mm,
value in male goose with a mean of 87.0 mm n =27), whereas the female PWF has a mean length
+0.2SD (range =86.7 to 87.2 mm, n =27) than the of 101.21 mm +0.06SD (range =101.13 to 101.31
female with a mean value of 86.8 mm +0.2SD mm, n =15). The PWF length was found
(range = 86.5 to 87.0 mm; n =15). During monsoon statistically significant at both 99% and 95%CL (p
season, the male goose has a greater NC with a <0.01, Students t-test) between the male and
mean of 87.0 mm +0.2SD (range =86.7 to 87.2 mm, female. During winter season, the mean PWF
n =13) than the female goose of the same season length measured in the male was 107.2 mm
with a mean NC of 86.8 mm +0.2SD (range =86.5 +0.07SD (range =107.07 to 107.31 mm, n =14),
to 87.0 mm, n =6). whereas in female goose the mean PWF length
i. Wing span (WS) remains at 101.23 mm +0.07SD (101.13 to 101.35
The mean WS of the Cotton Pygmy-goose was mm, n =9). During monsoon season, the mean PWF
found to be 425.3 mm +0.7SD (range 424.0 to was found to be 107.2 mm +0.07SD (range =107.07
426.0 mm, n =42, Table 5.1, Figure 5.1) during the to 107.31 mm, n =13) and 101.2 mm +0.05SD
study period. The male goose have a mean WS of (range =101.13 to 101.25 mm, n =6) respectively in
425.278 mm +0.7SD and female goose with 425.26 male and female gooses. The PWF length
mm +0.7SD (range =424.0 to 426.0 mm in both difference was found statistically significant in the
sexes, n =27 & 15). The mean WS of male during male and female at 99%CL (p <0.01, Students
winter was found to be 425.428 mm +0.6SD (range Paired t-test) during winter and monsoon season.
=424.5 to 426.0 mm, n =14), whereas the female The length-wise arrangement of the PWF of male
has a mean WS of 425.388 mm +0.8SD (range was found as PWF1< PWF11< PWF10 < PWF9 <
=424.0 to 426.0 mm, n =9) in the same season. PWF8 < PWF7 < PWF2 < PWF6 < PWF5 < PWF3
Again, during monsoon the male goose has a mean < PWF4 (Table 5.2 & Figure 5.2). A more or less
WS of 425.11 mm +0.7SD (range =424.0 to 426.0 similar arrangement was found in PWFs of female
mm, n =13) and female has 425.08 mm +0.7SD goose with a difference in PWF2 and PWF6, where
(424.0 to 426.0 mm, n =6). PWF6 < PWF2.
j. Wing length (WL) The mean SWF length was found to be 83.1
The mean WL (flattened) in case of both the mm +0.06SD (range =80.5 to 85.7 mm, n = 42).
sexes of Cotton Pygmy-goose recorded was 169.2 The male goose have longer SWF length with a
mm +0.6SD (range =168.0 to 171.0 mm, n =42, mean 85.6 mm +0.06SD (range 80.6 to 85.6 mm, n
Table 5.1, Figure 5.1). The male goose has a mean =27) than the female goose with a mean 80.7 mm
WL of 169.3 mm +0.85SD (range =168.0 to 171.0 +0.05SD (range 80.6 to 80.8 mm, n =15)
mm, n =27), whereas the female has a mean WL of irrespective of seasons. During monsoon, the male
169.1 mm +0.92SD (range =168.0 to 171.0 mm, n has a mean SWF length of 85.6 mm +0.05SD
=15). The WL of the female goose during the (range =85.55 to 85.64 mm, n =13), whereas the
monsoon season was measured with a mean WL of female has a mean length of 80.6 mm +0.03SD
169.3 mm +1.1SD (range =169.0 to 171.0 mm, n (range 80.6 to 80.7 mm, n =6). During winter the
=6), whereas in male the same remains at 169.0
mm +0.7SD (range =168.0 to 170.0 mm, n =13).
During winter season, the mean WL of the male
goose was found to be 169.5 mm +0.9SD (range
=168.0 to 171.0 mm, n =14) and in female the
mean WL was found as 168.9 mm +0.8SD (range
=168.0 to 170.0 mm, n =9).
k. Wing feathers (WF) & wing feather length
(WFL)
(a) (b)
The number of WF in both male and female
Cotton Pygmy-goose was 25 with 11-primaries Plate 2 Secondary wing feathers of (a) male & (b)
female CPG (from inner to outer)
male goose has a more or less similar mean SWF monsoon season the male goose have greater SFL
length with the monsoon season (mean: 85.6 mm with a mean SFL of 42.2 mm +0.3SD (range =41.9
+0.07SD, range =85.46 to 85.63 mm, n =14), to 42.5 mm, n =13) than the winter ones with a
whereas in the female goose the same remains at mean SFL of 42.0 mm +0.1SD (range =41.9 to 42.2
80.5 mm +0.07SD (range =80.52 to 80.75 mm, n mm, n =14). The scapulars found significantly
=9). The length-wise arrangement of the SWF in longer in males by 0.27 mm +0.13SD.
male was found to be SWF14 <SWF9 <SWF10 m. Tarsus length (TL)
=SWF13 < SWF8 < SWF7 < SWF6 < SWF5 < The mean TL in Cotton Pygmy-goose was
SWF4 <SWF1 <SWF3 <SWF11 <SWF2 <SWF12, found to be 29.1 mm +0.1SD (range =28.9 to 29.3
while in female the arrangement was found to be mm, n =42, Table 5.1, Figure 5.1). The mean TL in
SWF14 <SWF7 <SWF9 < SWF6 < SWF10 < both sexes of the goose was found to be 29.1 mm
SWF5 <SWF4 < SWF8 < SWF13 < SWF3 <SWF2 +0.1SD with a range between 29.0 to 29.2 mm
<SWF1 <SWF11<SWF12 (Table 5.3 & Figure (male: n =27; female: n =15).
5.2). The SWF length was found statistically n. Hind toe length (HTL)
significant at both 99% CL (p <0.01, Students t- In Cotton Pygmy-goose the mean HTL or
test) between the male and female. first toe length was found to be 8.1 mm +0.09SD
l. Scapular feathers (SF) and scapular feather (range =7.9 to 8.3 mm, n =42, Table 5.1, Figure
length (SFL) 5.1). The male goose have a slightly longer mean
There are 6 (six) SF in both male and female HTL with 8.2 mm +0.09SD (range 8.0 to 8.3 mm, n
Cotton Pygmy-goose with mean length of 42.0 mm =27) than the female goose which have a mean
+0.25SD (range =41.7 to 42.6 mm, n =42, Table HTL of 8.06 mm +0.09SD (range =7.9 to 8.2 mm, n
5.1). The male goose has a mean SFL of 42.2 mm =15) irrespective of the seasons of a year. During
+0.3SD (range =41.9 to 42.6 mm, n =27) whereas winter, the mean HTL of male goose was found to
the female goose has a mean SFL of 41.9 mm be 8.1 mm +0.1SD (range =8.0 to 8.3 mm, n =14),
+0.1SD (range =41.7 to 42.1 mm, n=15) whereas a mean length of 8.0 mm +0.08SD (range
irrespective of the seasons of a year. During =7.9 to 8.2 mm, n =9) was observed in female
goose during the same season. The male has a
greater HTL than the females during the monsoon
(male: mean 8.2 mm + 0.08SD, n =13; female:
mean 8.1 mm +0.1SD, n =6).
o. Inner toe length (ITL)
In Cotton Pygmy-goose the mean ITL or
second toe length was found to be 31.7 mm +0.1SD
(range =31.5 to 32.0 mm, n =42, Table 5.1, Figure
5.1). The male goose have a slightly longer mean
(a) (b) ITL with 31.8 mm +0.2SD (range 31.4 to 32.0 mm,
Plate 3 Rectrices of (a) male & (b) female CPG from n =27) than the female goose which have a mean
right to left) ITL of 31.6 mm +0.1SD (range =31.5 to 31.8 mm,
196 Journal of Research in Biology (2011) 3: 191-201
Upadhyaya et al.,2011
was found to be 16.5 mm +0.2 (range 16.2 to 16.8 with mean TaL 64.4 mm +0.6SD (range =63.2 to
mm; n =42, Table 5.1) irrespective of sexes and 65.3 mm, n =9). The numbers of TF or rectrices in
seasons. The male and female goose have a more Cotton Pygmy-goose vary in both sexes. The male
or less similar measurement of OWD with a mean has 12 TF, whereas the female has 14 in numbers.
of 16.5 mm +0.2SD in both (range =16.2 to 16.8 The mean TFL was found to be 74.2 mm +0.08SD
mm, n= male-27 & female-15) irrespective of the (range =73.36 to 75.07 mm, n =42, Table 5.1)
seasons of a year. During winter season, the mean irrespective of sexes and seasons. The mean TFL of
OWD of male and female goose was found to be the male was 73.5 mm +0.09SD (range =73.4 to
16.5 mm +0.2SD (range =16.2 to 16.8 mm, n =male 73.6 mm, n =27) and in female it remains at 74.9
-14 & female-9). Again during monsoon season, the mm +0.08SD (range =74.8 to 75.1 mm, n =15).
mean OWD of male and female goose was found to During winter, in male goose the mean TFL found
be 16.5 mm +0.2SD and 16.4 mm +2SD (range: was 73.5 mm +0.09SD (range =73.4 to 73.7 mm, n
male =16.2 to 16.8 mm, n =13; female =16.2 to =14) and in female the mean value found to be 75.0
16.7 mm, n =6). mm +0.08SD (range =74.9 to 75.1 mm, n =9).
t. Tail length (TaL), tail feathers (TF) or Again, the male goose studied during monsoon
rectrices and tail feather length (TFL) shows a mean TFL of 73.5 mm +0.09SD (range
The mean TaL in Cotton Pygmy-goose was =73.4 to 73.6 mm, n =13), whereas the female
found to be 64.6 mm +0.8SD (range =63.2 to 65.9 goose has a mean TFL of 74.9 mm +0.09SD (range
mm, n =42, Table 5.1, Figure 5.1). The male has a =74.9 to 75.1 mm, n =6).
longer tail with a mean TaL of 64.6 mm +0.8SD Twenty pairs of body parameters were
(range =63.4 to 65.9 mm, n =27) than the female studied and relationship was correlated which
with a mean TaL 64.5 mm +0.5SD (range =63.2 to shows strong as well as negative relationship
65.3 mm, n =15). The goose studied during winter among them, and the value of r ranges between -
season has greater TaL with mean 64.6 mm +0.6SD 0.9 to 0.9 (Table 5.4 & Figure 5.3). A strong
(range =63.2 to 65.9 mm, n =23) than the goose relationship was observed in case of female (r=0.8,
studied during monsoon season with a mean 64.5 Pearson Correlation) between BoW and BoL, while
mm +0.6SD (range =63.2 to 65.9 mm, n =19). The in the case of male a less strong correlation (r=0.2)
male goose captured during winter has longer tail was observed. It shows a higher tendency of
with mean TaL of 64.7 mm +0.7SD (range =63.4 to increase of body weight with an increase in BoL.
65.9 mm, n =14) than the female of the same season There exists a strong correlation between BoL and
Table 4. The co-efficient of correlation (r) among various body parameters of CPG
(male & female)
Parameters r in male r in female
1. Body wt. & Body length 0.2 0.8
2. Body length & Neck length 0.7 0.8
3. Body length & Bill length 0.9 0.1
4. Bill length & Bill depth 0.5 0.4
5. Body length & Middle toe length 0.7 -0.1
6. Tarsus & Middle toe length 0.1 0.5
7. Body length & Tail length 0.5 0.2
8. Wing length & Tail Length -0.7 -0.4
9. Wing feather length & Tail feather length -0.7 -0.1
10. Wing length & Body length -0.8 0.5
11. Bill length & Bill width -0.6 0.1
12. Wing Expansion & Body length -0.8 -0.7
13. Bill depth & Bill width -0.2 -0.1
14. Tarsus length & Hind toe length -0.2 -0.2
15. Tarsus length & Inner toe length 0.1 0.2
16. Hind toe length & Middle toe length -0.3 0.1
17. Tarsus length and Outer toe length -0.5 0.1
18. Outer toe length & Middle toe length 0.4
19. Hind toe length & Outer toe length 0.5 -0.7
20. Outer toe length & Inner toe length 0.5 0.9
Plate 5 An adult CPG (female) during measurement Plate 6 A pair of adult CPG recovered from
(in inset: an adult male) a poacher of Kadamani area.
NL in both male and female (r=0.7 & 0.8 irrespective of their sex except a male bird which
respectively) and a less strong correlation between may just arrived to the area and was trapped for the
the BoL and BL in female (r=0.1). The NL is study. The greater weight in the birds of the
highly related with the BoL and vice-versa, while vegetation cover area may be due to recent feeding
the tendency is lower in case of the male. A on the vegetation available in the wetland. The
moderately strong correlation was observed in Cotton Pygmy-goose studied from Borsola area
female (r=0.5) between TL and MTL. The co- were found greater weight than the goose captured
efficient of correlation between BoL and TaL was from other areas during the study period. The
found strong in male (r=0.5) and in female (r=0.2), reason might be the availability of food plants in the
whereas the WS and BoL in both male (r=-0.8) and wetlands of Borsola area.
female (r=-0.7) are oppositely related and increase The sex-wise differences of body length,
in WS decreases the BoL. An oppositely related plumage characteristics were found to be prominent
WFL and TFL were found in male (r = -0.7) and in the present context of the study. Deviations of
female (r= -0.1). Similarly the BD and BW in both the body lengths were more like those predicted by
male and female were oppositely related (r = -0.2 & the competition hypothesis in wetlands with low
-0.1 respectively). food abundance than in the wetlands with high food
abundance. The similar results were also discussed
DISCUSSION by Pys et al. (1994).
The present morphomatric study on forty T h e f u nct i on al s i gni f i c an ce of
two Cotton Pygmy-gooses, the average BoW of morphomatric study shows that due to differences
male and female is found to be 221.2 gm and 227.1 in neck and body length the Cotton Pygmy-goose
gm respectively. The study shows a low body use different feeding methods and depths for
weight of the Cotton Pygmy-goose in comparison feeding. The present findings were further
to other goose. A lower body weight of the supported by the works of Pys (1983a, 1983b,
Nettapus sp. > 500 gm has been confirmed. The 1986 & 1987). The wing span and the flattened
female Cotton Pygmy-goose was found heavier wing length are found quite correlative in response
during the monsoon season than the male. This to the flight adaptation. The relationship of wing
might be either due to excessive feeding for the span, wing length and tarsus length with the body
developing eggs during monsoon season or food weight is also supported by the works of Green et
reservation which may occur for future use during al. (2001). No relation was observed in between the
incubation of the eggs. The present investigation outer web distance and inner web distance. Fox et
also reveals that the female has slightly greater al. (1992) found significance differences between
body weight than the male bird during breeding male and female adult teal in tarsus and wing
season prior to egg laying. The weight of the birds length. The present findings are also more or less
congregated in vegetation covers area found to be supported by the findings of Moore and Battley
greater than the birds found in open area (2003) in Anas chlorotis.
Authors: ABSTRACT:
1,2
Alisi P. Nc,
1
Buseri FI and
3
Alisi CS.
Haeomatological and oxidative stress parameters were measured in Pre-
Institution: eclamptic [PE] women attending clinic in owerri South Eastern Nigeria. The effect of
1
Department of medical Pre-eclampsia on these parameters was assessed by juxtaposition with similar indices
Laboratory Science in normal pregnant and normal non-pregnant women. Result showed that pre-
Rivers State University of eclampia resulted in significant decrease (P< 0.05) in Lymphocyte count. There was a
Science and Technology significant increase in total white blood cell count, neutrophil count and mixed
Port Harcourt, Nigeria differential count. Pre-eclamptic resulted in significant increase (P<0.05) in
erythrocyte lipid peroxidation, met-haemoglobin concentration and percentage
2
Federal medical centre hydrogen peroxide-induced haemolysis. The result indicate that oxidative stress is a
PMB 1010 Owerri, Nigeria significant contributor in the pathogenesis and white cell changes seen in pre-
3
eclampsia.
Department of
Biochemistry Federal
University of Technology
PMB 1526 Owerri.
Dates:
Web Address: Received: 21 May 2011 /Accepted: 26 May 2011 /Published: 18 Jul 2011
http://jresearchbiology.com/
Documents/RA0033.pdf.
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.
basis of maximum absorbance (100%) in the increase in prenatal mortality and its socioeconomic
aliquots of erythrocyte completely haemolysed in impact on developing countries is huge, its
distilled water. pathophysiological changes include elevated
Measurement of Methaemoglobin (Hi) in blood systemic vascular resistance, generalized
Methaemoglobin has a maximum absorption vasoconstriction, and activation of the coagulation
at 630nm. When cyanide is added this absorption cascade, all of which may be explained by
band disappears and the resulting change in disruption of normal maternal endothelial function
absorbance is directly proportional to the (Lpez-Jaramillo et al., 2001).
concentration of Hi. Total Haemoglobin in the Effect of pre-eclampsia on hydrogen
sample is then measured after complete conversion peroxide induced haemolysis in erythrocytes (figure
to HiCN by the addition of ferricyanide-cyanide 1), shows that erythrocytes from pre-eclamptic
reagent using the method of Evelyn and Malloy patients underwent more lysis than erythrocytes
(1938). from the normal pregnant control (PC) and the non-
Briefly, 0.2mls of blood was lysed in a pregnant control (NPC). It is known that the
solution containing 4ml of buffer and 6ml of increased lysis results from oxidative damage to the
nonidet P40. The lysate was divided into two equal erythrocyte membrane, causing a decrease in
volumes (A and B). The absorbance of A was membrane fluidity and reducing its ability to
measured in a spectrophotometer at 630nm (D1). 1 withstand osmotic changes. Hydrogen peroxide
drop of KCN solution was added and absorbance participates in the HaberWeiss reaction: O2 +
was measured after mixing (D2). 1 drop of K3Fe H2O2 O2 + OH + OH. Increase in the H2O2
(CN)6 was added to solution B and after 5mins induced haemolysis in pre-eclampsia could indicate
absorbance was measured at the same wavelength a lesser ability to neutralize hydrogen peroxide
(D3). Then 1 drop of KCN was added to solution to (H2O2 + 2H+ + 2e 2H2O). Our observation
B and absorbance was measured after mixing (D4). could mean that there is a decreased antioxidant
Measurements were made against a blank capacity in pre-eclampsia. Increase in osmotic lysis
containing buffer and detergent in the same have been reported in pre-eclampsia in the united
proportion as present in the sample. % kingdom and the Netherland (Courinne et al., 1998;
methaemoglobin concentration was calculated thus: Armutcu, 2005).
Excessive formation of methaemoglobin was
% Methaemoglobin = D1-D2 C 100 seen in the erythrocytes of the preclamptic patient
D3-D4 (figure 2) in comparison with the control groups.
Measurement of Lipid Peroxidation This could be as a result of the oxidation of
Lipid peroxidation in the erythrocyte was erythrocyte protein (haemoglobin) which leads to
determined spectrophotometerically by assessing accumulation of methaemoglobin. The above is
the level of thiobarbituric acid reactive substance similar to accumulation of methaemoglobin seen in
(TBARS) as described by Liu et al. (1990).
Absorbance was measured at 532 nm using a 100
% Hydrogen Peroxide induced hemolysis
oxidative damage by drugs and chemical agents. The total white cell count (table 1) in pre-
Increases observed in methaemoglobin eclamptic women was significantly elevated than
concentration in pregnancy and pre-eclampsia the NPC. A significant increase in total white cell
implied a shift in pro-oxidant/antioxidant count was also found in PE when compared to PC
equilibrium in favour of the pro-oxidant, resulting women. This observation implied that pre-
in stress. Increased concentration has been eclampsia further exacerbated leukocytosis as
postulated in diseases associated with oxidative pregnancy is universally known to induce
stress (Dacie and Lewis 2006). leukocytosis. This is in agreement with Balloch
In this study, erythrocyte lipid peroxidation and Cauchi (1993). Schuiling et al., (2000) found
(LPO) as seen from the malondialdehyde an increased activated white blood cell in
concentration were significantly higher in the blood preeclampsia than in normal pregnancy, while
of pre-eclamptic women (figure 3) than the Sargent et al., (1998) found an increase in activated
controls. However, the RBC LPO in the pregnant white blood cells both in normal pregnancy and
control was also higher than the non-pregnant preeclampsia which was more in preclampisa
control. This shows that during uncomplicated resembling sepsis condition. On the other hand
pregnancy, there is an increased production of pro- Ceyhan et al., (2006) and Heilman et al., (2004)
oxidants that is balanced by the synthesis of recorded in their study an increase in total white
antioxidants (Scholl et al., 2005). Sims et al., cell count in preeclampsia that was not statistically
(1998) also reported that serum lipid peroxide significant. Correlation has been found to exist
concentrations in pregnant subjects were between stress conditions and higher total white cell
significantly higher than those in non-pregnant count (Allsop et al 1992). This could lead one to
subjects. The significant variation seen in the pre- say that white blood cells are activated in pre-
eclamptic group and pregnant control is suggestive eclampsia and that pre-eclampsia may be partly an
that pre-eclampsia resulted from an imbalance inflammatory disorder since the activation of white
between pro-oxidant production and probably the cells were similar to that seen in inflammatory
antioxidant defenses. This imbalance could disorder (Sargent et al.,1998).
contribute to vascular dysfunction (Shaarawy et al., The percentage lymphocyte in pre-eclamptic
1998). The nature and extent of oxidative injury women was significantly decreased when compared
contributing to vascular dysfunction would depend to the normal pregnant controls. The result of this
on the concentration of free radicals/ reactive study agreed with reports from Israel (Lurie et al.,
oxygen species. Increases in lipid peroxidation 1998) but reports from Louisiana (Canzoneri et al.,
products like malondialdehyde correlates directly 2009) was different, no statistical significant
with increased imbalance in pro-oxidant/antioxidant difference was seen in lymphocyte between the PE
concentrations. and normal PC. The variation in this parameter
could come from the increase in the percentage
4.00
4
3.00
Percentage (%) Methaemoglobin
3
MDA (mol/l)
2.00
2
1.00
1
0.00
0 NPC PC PE
NPC PC PE
Ser i es1
Eskenazi B, Fenster L, Sidney S and Elkin EP. Sargent IL, Sacks GP, Studena K and Redman
1993. Fetal growth retardation in infants of CWG. 1998 Normal pregnancy and preeclampsia
multiparous and nulliparous women with both produce inflammatory changes in peripheral
preeclampsia. American Journal Obstetrics and blood leukocytes akin those of sepsis. American
Gynecology 169:1112-1118. Journal of Obstetrics and Gynecology 179:80-86
Evelyn KA and Malloy HT. 1938. Scholl OT, Leskiw M, Chen X, Sims M and Stein
Microdetermination of oxyhaemoglobin, PT. 2005. Oxidative stress, diet, and the etiology of
Methaemoglobin and sulfhaemoglobin in a single pre-eclampsia. American Journal of Clinical
sample of blood. Journal of Biological Chemistry Nutrition 81:1390-6.
126:655.
Schuiling MM, Faas MM, Gerard A, Linton,
Goldenberg RL and Rouse DJ. 1998. Prevention Elizabeth A, Sargent IL, Redman C WG. 2000.
of premature birth. New England Journal of Activation of peripheral leukocytes in rat pregnancy
Medicine 339:313-320. and experimental preeclampsia American Journal of
Obstetrics and Gynecology 182(2):351-357.
Heilmann L, Rath W, Pollow K. 2004.
Hemoconcentration and pre-eclampsia Se Imseek M, Nairogi Lu M, Se Imseek M, Ce
Clinical Hemorheology and Microcirculation 31 Ay M, Aksakal M and Kumru S. 1998. Blood
(1):49-58. Plasma Levels of Lipoperoxides, Glutathione
Peroxidase, Beta Carotene, Vitamin A and E in
Liu J, Edamatsu R, Kabuto H, Mori A. 1990. Women with Habitual Abortion Cell Biochemistry
Antioxidant action of Guilingji in the brain o f and Function. 16:227-231.
rats with FeCl3 induced epilepsy. Free Radical
Biology and Medicine 9, 451-454. Sengupta A, Guha SK, Anand S, Jayaraman G,
Biswas P and Rehana HMK. 1995. Can
Lpez-Jaramillo P, Casas JP and Serrano N. conventional biomedical investigative techniques
2001. Preeclampsia: from epidemiological unfold the mysteries of uteroplacental system in
observations to molecular mechanisms. Brazilian pregnancy? A critical study. In: The proceedings of
Journal of Medical and Biological Research 34 RC-IEEE-EMBS, USA. NewDelhi:CBME-IIT/
(10):1227-1235. AIIMS 1.83-1.84.
Authors: ABSTRACT:
Alisi CS, Nwaogu LA,
Ibegbulem CO and
Inhibition of total dehydrogenase enzyme activity in pathogenic gram
Ujowundu CU.
positive and gram negative micro organisms exposed to methanol extract of
Chromolaena odorata was used as an index for assessment of its antimicrobial
Institution: activity. Assay of total dehydrogenase enzyme activity was done in the test organisms
Department of Biochemistry, (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli) using 2,3,5-
Federal University of triphenyltetrazolium chloride (TTC) as an artificial electron acceptor which was
Technology, reduced to the red-coloured triphenyl-formazan. Response of the bacterial isolates
PMB 1526 Owerri, Imo varied with extract concentration. Dehydrogenase activity was progressively inhibited
State, Nigeria. in a logistic dose-response fashion. The gram negative Escherichia coli responded
more markedly than Pseudomonas aureginosa and gram positive Staphylococcus
aureus. Inhibitory concentrations (IC50) of the methanol extract against Escherichia
Corresponding author: coli, Staphylococcus aureus, and Pseudomonas aeruginosa were 208.49 g/ml,
Alisi CS 1361.01 g/ml, and 903.08 g/ml respectively. Preliminary phytochemical screening
of the extract gave positive reactions for alkaloids, flavonoids, tannins, 4-
hydroxybenzoic acid, and glycosides. These phytochemicals may be responsible for
Email: the observed inhibition of total dehydrogenase enzyme activity that translates to
silverpresh@yahoo.com antimicrobial action in these pathogenic organisms.
Keywords:
Oxidoreductases, toxicity, enzyme inhibition, wound isolates, phytochemicals,
and bacterial response.
Article Citation:
Web Address:
http://jresearchbiology.com/
Alisi CS, Nwaogu LA, Ibegbulem CO and Ujowundu CU.
Documents/RA0016.pdf. Antimicrobial Action of Methanol Extract of Chromolaena Odorata-Linn is
Logistic and Exerted by Inhibition of Dehydrogenase Enzymes.
Journal of research in Biology (2011) 3: 209-216
Dates:
Received: 05 May 2011 /Accepted: 09 May 2011 /Published: 18 Jul 2011
Ficus Press.
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.
odoratum L.), a perennial belonging to the plant 2006), and heterotrophic (Nweke, et al., 2007)
family Asteraceae (=Compositae), is a diffuse, bacteria. We had earlier also assessed toxicity of
scrambling shrub that is mainly a weed of antimicrobial agent to pathogenic bacteria using the
plantation crops and pastures of Southern Asia and dehydrogenase assay (Alisi, et al., 2008; Nwaogu,
Western Africa. It is a weed of 13 crops in 23 et al., 2008; Nwaogu, et al., 2007).
countries and has been described as the worlds Results of an earlier study showed that the
worst weed (Holme et al., 1977). This common extract of the leaves of C. odorata inhibited the
plant called Siam weed is known among the Igbos growth of some bacteria (Alisi and Onyeze, 2009).
of the South-Eastern Nigeria as: Elizabeth, Inhibition of dehydrogenase enzymes in pathogenic
Independence leaf, Enugu plantation weed, or pure microbial cultures by methanol extracts of C.
Awolowo weed. Chromolaena odorata is used as odorata has not been demonstrated. This work is
a gargle for sore throat and cold. Traditionally, therefore aimed at studying the inhibition of total
fresh leaves or a decoction of C. odorata have been dehydrogenase enzymes in pathogenic pure
used throughout Vietnam for many years as well as microbial cultures exposed to methanol extract of
in other tropical countries for the treatment of leech Chromolaena odorata.
bite, soft tissue wounds, burn wounds, skin
infection and dento-alveolitis (Le, 1995). MATERIALS AND METHODS
Decoctions of leaves have been used by traditional Plants
healers in the South-Eastern Nigeria for the Fresh aerial parts of C.odorata were
treatment of liver diseases. collected from Egbu and Ihiagwa in Owerri North
Several subclasses of flavonoids have been and Owerri West local Government areas of Imo
isolated from C. odorata extracts. The phenolic State respectively. Plant was authenticated by
acids present in ethanol extract of C.odorata are Professor S.E.Okeke, a plant taxonomist, of the
protocatechuic, p-hydroxybenzoic, p-coumaric, Department of Plant Science and Biotechnology,
ferulic and vanillic acids. The complex mixtures of Imo State University Owerri, Imo State. Voucher
lipophilic flavonoid aglycones present in C.odorata specimen is deposited in the authors laboratory.
included flavanones, flavonols, flavones and Extract Preparation
chalcones. C. odorata also contains high The aerial part of C. odorata was shed dried
concentrations of amino acids (Phang, et al., 2001). at room temperature and reduced to a coarse
The use of the total dehydrogenase assay has been powder in a mill (Kenwood BL357). The powder
used as a tool in probing response of was extracted with methanol. The extract was
microorganisms to antibacterial agents (Alisi et al., recovered by distillation under reduced pressure at
2008) and is recognized as a useful indicator of the 49oC in a rotary evaporator-Buchi rotavapour
overall measure of the intensity of microbial, (Switzerland). The extracts were then dried to solid
metabolism (Tabatabi 1982; von Marsi and forms in vacuum desiccators, and stored in a freezer
Schinner, 1991). This method is preferred over (<-4.0 0C).
Table 1: Phytochemical constituents of Chromolaena.odorata methanol extracts
4-HBA
(p-OH-
Cardiac Steroidal benzoic
Alkaloids Flavonoids Tannins Saponins Glycosides glycosides aglycone Protein acid)
Dry
plant + + + + + + + + +
MECO + + + - + + + + +
Isolation of bacterial strains and culture exponential phase in nutrient broth (Lab M) on a
conditions Marrienfeld rotary incubator (150 rpm) at room
Pathogenic bacteria (Pseudomonas sp., temperature (28 2oC). The cells were harvested by
Staphylococcus sp., and Escherichia sp. were centrifugation at 4000 rpm for 10 min. Harvested
obtained from degenerated wound. Isolates were cells were washed twice in deionised distilled water
purified on nutrient agar (Fluka) plates and and re-suspended in water. The re-suspended cells
characterizations were done using standard were standardized in a spectrophotometer to an
microbiological methods. Identifications to the optical density of 0.70 at 420 nm. The dry weights
generic level followed the schemes of Holt et al. of the standardized cells were determined by drying
(1994). The bacterial strains were grown to mid volumes of cell suspension to constant weight in an
Table 2: Showing the threshold inhibitory concentrations of methanol extracts of C.odorata against the total
dehydrogenase activity (DHA) of some wound isolates (Escherishia.coli, Staphylococcus.aureus, and
Pseudomonas.aureginosa)
Inhibitory Concentrations Against Wound Isolates
MECO(g/ml)
IC20 IC50 IC70 IC80 IC100
Escherishia coli 19.15 208.49 * 4736.25 9755.00
Staphylococcus aureus 111.11 1361.01 * ND ND
Pseudomonas aureginosa 206.27 903.08 1843.57 2605.12 5473.75
1.5 2
Log % Inhibition
1.75
1.25 1.5
1.25
Journal of Research in Biology
1 1
0.75
0.75 0.5
0.25
10 100 1000 10000
0.5
0.25
0 500 1000 1500 2000
MECO (ug/ml)
Figure 1b: Plot of Log percentage inhibition of DHA
against graded concentration of methanol extract of
chromolaena odorata
Staphylococcus sp.
A
1.1 1.1
1
0.4
0.7 0.01 0.1 1 10 100 1000 10000
0.6
0.5
0.4
0 500 1000 1500 2000
B ETECO(ug/ml)
60
50
% INHIBITION OF DHA
40 60
50
30
40
30
20
20
10 10
0
0 -10
0.01 0.1 1 10 100 1000 10000
-10
0 500 1000 1500 2000
C ETECO(ug/ml) D
1.5 1.75
1.5
1
Log % INHIBITION
r-PARAMETER
1.25
0.5 1
0.75
0
0.5
-0.5 0.25
0 500 1000 1500 2000 0 500 1000 1500 2000
ETECO(ug/ml) ETECO(ug/ml)
MECO (g/ml)
Fig.2 : Inhibition of dehydrogenase activity (mg Formazan/mg cell dry weight/hr) in Pathogenic (wound
isolates) bacteria(Staphylococcus. sp.) by methanol extract of Chromolaena odorata. Plots (A,B,C,and D) show
dehydrogenase activity, percentage inhibition of DHA, gamma parameters and log %inhibition of DHA on y-
axis respectively against graded concentrations of extract on x-axis.
amount of formazan produced was determined from inhibition data generated are fitted into the model
a standard dose-response curve [0 - 20 g/ml TPF equation (2) which is a logistic dose response
(Sigma) in amyl alcohol; y = 0.0487x; R2 = equation. The parameters were estimated by
0.9977]. Dehydrogenase activity (DHA) was iterative minimization of least squares using
expressed as milligrams of TPF formed per mg dry Levenberg-marquardt algorithm (Table curve 2D
weight of cell biomass per hour. systat USA) Marquardt (1964). The data of %
Data Analysis inhibition fitted into equation (2) were used to
Percentage Inhibition of dehydrogenase evaluate the toxicity thresholds IC5, IC20, IC50, IC70,
activity by MECO was calculated relative to the IC80, IC100 which are the concentrations of the
control as shown in equation (1) (table 1). The extracts that inhibited 5%, 20%, 50% 70%, 80%
Pseudomonas sp
and 100% of controls respectively. Where IC100 was
0.9
A 0.9 not determinable (ND) by fitting Eqn (1) into the
model (Eqn 2), Equation (1) was transformed to
DHA (mg Formazan/mg cell dry wt/hr)
0.8
0.3
parameter plot where data gave high R2-value with
0.5 0.2
0.01 0.1 1 10 100 1000 10000 Eqn 4. Data whose IC100 was non-determinable
using Equation 2 and 3, were fitted into an inverse
0.4
Log(x) polynomial equation (Eqn 5). By solving for
x in Eqn 5, IC100 (the concentration at which MECO
0.3
will exert total inhibition against the tested
0.2 organism) was calculated.
0 500 1000 1500 2000
MECO (ug/ml)
B RESULTS AND DISCUSSION
80
The plant C.odorata was found to contain
Flavonoids, Tannins, Saponins, Glycosides,
%Inhibition of DHA in Pseudomonas
70
Steroidal aglycones, Alkaloids and 4 -
60 hydroxybenzoic acid. The methanol extract
50
however did not show a positive reaction for
80
saponins (Table 1). These phytochemicals have
70
40 60
been found to have medicinal properties and health
50
promoting effects (Raza and John, 2007; Salah et
30
40 al., 1995; Del-Rio et al., 1997; Okwu, 2004; Liu,
20
30 2004). The use of C.odorata in ethno-medical
20
practice may be due to the medicinal effects of
10 10
these phytochemicals.
0
0.01 0.1 1 10 100 1000 10000
Methanol extract of C.odorata inhibited
0
0 500 1000 1500 2000 dehydrogenase activity in the organisms in a
MECO (ug/ml) logistic dose dependent manner. Inhibition of
C
3
dehydrogenase activity in Staphylococcus aureus,
2.5 and Pseudomonas aerugenosa followed a logistic
dose response abcd model (Eqn 1) while E.coli,
followed weilbullcum abcde model Eqn (3) and
r-PARAMETER
2
(Eqn 5).
1.5 Threshold inhibitory concentrations of the
extracts (Table 2) showed that Pseudomonas
1
auregenosa responded gradually but steadily. At
0.5
lower concentrations, the extracts exerted stronger
inhibitory effect on the dehydrogenase activity of
0 Escherishia coli and Staphylococcus aureus than
0 500 1000 1500 2000
MECO (ug/ml) Pseudomonas auregenosa. The rate of inhibition of
total dehydrogenase enzyme activity in Escherishia
Fig.3: Inhibition of dehydrogenase activity (mg coli and Staphylococcus aureus was not sustained
Formazan/mg cell dry weight/hr) in Pathogenic as Pseudomonas auregenosa responded more at
(wound isolates) bacteria(Pseudomonas sp..) by higher concentrations of C.odorata extracts, making
methanol extract of Chromolaena odorata. Plots IC100 in Escherishia coli and Staphylococcus aureus
(A,B,and C) show dehydrogenase activity,
percentage inhibition of DHA and gamma
non-determinable.
parameters on y-axis respectively against graded The gamma parameter model (Eqn 4) gave a
concentrations of extract on x-axis. strong linearization of percentage inhibition of
DHA by methanol extract of C. odorata against
214 Journal of Research in Biology (2011) 3: 209-216
Alisi et al.,2011
Table 3: Showing models and equations for dehydrogenase inhibition
with pearson correlation coefficient in the pathogenic organisms.
Organisms Model Equation R2-value
WEIBULLCU Eqn (3) 0.9969
Escherishia coli M (abcde)
inverse Log(x) Eqn (5) 0.9989
polynomial
Journal of Research in Biology
Pseudomonas aerugenosa. This makes this the Ibegbulem CO. 2008. Inhibition of dehydrogenase
model of choice in the determination of IC50 in activity in pathogenic bacteria isolates by aqueous
Pseudomonas aerugenosa. All equations used in the extracts of Musa paradisiaca (Var Sapientum). Afr.
modelling of the results gave high correlation Journ. Biotech. 7(12):1821-1825.
coefficient (R2 0.99), with very low Fit standard
errors. Inhibition of dehydrogenase activity in the Aziz NH, Frag SE, Mousa LA and Abo-Zald
wound isolates showed antimicrobial activity. The MA. 1998. Comparative antimicrobial and
extract was toxic to the organism at all antifungal effects of some phenolic compounds.
concentrations and the nature of inhibition was Microbios 93(374):43-54.
logistic rather than hormetic (Table 3).
Extracts of the plant C. odorata have earlier DelRo A, Obdulio BG, Castillo J, Marin RR,
been shown to contain phenolic compounds (Phan, and Ortuno A. 1997. Uses and properties of citrus
et al., 2001). The presence of phenolic compounds flavonoids. J. Agric. Food Chem. 45:4505-4515.
in the plant C.odorata has been demonstrated by
earlier worker (Phan et al., 2001). Derivatives of 4- Evans WC. (ed) 2002. Trease and Evans
hydroxybenzoic acid are used as anagelsics and pharmacognosy: Edinburg:W.B.Saunders.
antimicrobial substances. Also, Tannins,
flavonoids, and saponins are known to have Holme GLR, Plucknet DL, Pancho JV and
antimicrobial activities (Aziz et al., 1998; Evans, Herberger JP. 1977. Chromolaena odorata, The
2002). Hydroxybenzoic acid found in the extract is Worlds Worst Weeds: Distribution and Biology:
known to possess antimicrobial activity. The The University Press of Hawaii.
secondary plant metabolites identified in this
extract may be acting synergistically to bring about Le TT. 1995. The 5th European Tissue Repair
the observed inhibition of dehydrogenase activity. Society Annual Meeting, Padova, Italy, (Abst. 30).
These extracts may actually be exerting their
antimicrobial activity via inhibition of Liu RH. 2004. Potential synergy of phytochemicals
dehydrogenase activity in the test organisms. in cancer prevention: mechanism of action. J. Nutr.
134:3479S- 3485S.
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Authors: ABSTRACT:
Kami Kaboosi In recent decades, specific big share of executive capabilities, capital and
activities of the Iranian government have been allocating to the development of
pressurized irrigation systems. Concluding these programs and specifying the
development status of these systems in different locations of country can be regarded
as a guideline to assign future strategies. Reliable information on irrigation methods is
important for determining agricultural water demand trends. Therefore, this research,
in addition to surveying of pressurized irrigation projects performed from 1990 to
Institution: 2007 in Golestan province; consider development trend and status of pressurized
Islamic Azad University, irrigation systems in this province. Based on the result, totally 1020 pressurized
Gorgan branch, irrigation projects with 22990.9 hectares area have been performed in Golestan
Gorgan, Iran. province, which have covered only 6.64 percent of irrigated lands area or 3.2 percent
of total agricultural lands area and shows the lack of pressurized irrigation
development. Comparing the pressurized irrigation development trend in Golestan
province with the country and their coordination shows that the most important
factor in the lack of pressurized irrigation development in Golestan province is general
policies on the country scale. The most important reason of very fluctuation in
developing of these systems is continuous changes in policies and instability of
Corresponding author: adopted policies while, technical, social, cultural and executive factors have the least
Kami Kaboosi effects. Results show more than 90 percentage of sprinkler irrigation systems area
allocated to cotton, soybean, wheat and barley. Also, because of petty landowner of
agricultural lands in Golestan province, area average of personal farms that equipped
with pressurized irrigation systems is very lesser than cooperative farms. This paper
present comprehensive conclusion of sprinkler and micro irrigation projects from
various aspects in Golestan province in Iran.
Email: Keywords:
kkaboosi@yahoo.com Development, Pressurized Irrigation, Iran.
Dates:
Received: 03 May 2011 /Accepted: 13 May 2011 /Published: 20 Jul 2011
Ficus Press.
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.
planning require reliable estimates of crop and condition, this province has important role in the
irrigation system combinations, which are important agricultural production of country. Based on
components of water budget. To update records on published statistical information by agriculture
irrigation methods used within each region, survey ministrys head-office of Statistics and Information
should been conducted. Then, survey data are (2005), total agricultural land area of Golestan
analyzed and compared with earlier surveys to province is 717594.6 hectares that is equal to 5.32
study how irrigation methods are changing and to percentage of total agricultural land area of country.
make projections of future changes for long-term
planning. MATERIALS AND METHODS
Iranian government has particularly To update records on pressurized irrigation
attentions to the development of pressurized methods, a survey was conducted. In Iran, farmers
irrigation systems in order to improve water use equip agricultural lands to pressurized irrigation
efficiency and water application efficiency. So, in systems by governmental loans. Therefore, data of
recent decades, specific big share of executive farms that equipped with pressurized irrigation
capabilities, capital and activities of the Iranian systems are available. Those are with Iranian
government have been allocating to the general office on pressurized irrigation. Pressurized
development of pressurized irrigation systems. irrigation office in Golestan province is a branch of
Concluding these programs and specifying the Iranian general office on pressurized irrigation. In
development status of these systems in different the first step, total information about pressurized
locations of country can be regarded as a guideline irrigation projects that performed from 1990 to
to assign future strategies. 2007 in Golestan province gathered from
To update Californias records on crops and pressurized irrigation office in Golestan province.
irrigation methods, the California department of Then, these data (projects) from the viewpoints of
water resources conducts survey every 10 years number and area of projects, variety of user, variety
during recent decades (Orang et al, 2008). of pressurized irrigation systems, crops, etc were
Therefore, Orang et al. (2005 and 2008) conducted analyzed. Finally, on the basis of results, it was
a survey of irrigation methods in California in 2001. analyzed development trend and status of
Kahlown and Kemper (2007) evaluated the pressurized irrigation in Golestan province.
performance of trickle systems installed in
Balochistan, Pakistan during 19822002 by field RESULTS
surveys, physical verifications and interviews with 1- Number and area of pressurized irrigation
farmers. projects
Akhavan Giglo (2006) and Akhavan Giglo Number and area of pressurized irrigation
and Kanooni (2008) presented the situation of projects that performed from 1990 to 2007 in
pressurized irrigation systems in private and Golestan province is present in table 1. As shown,
governmental agricultural lands of Ardebil totally 1020 pressurized irrigation projects with
province, Iran. 22990.9 hectare land usage were performed that are
The purposes of this research is twofold; (1) mostly sprinkler systems and only 6 percentage of
to survey of pressurized irrigation projects that the number of pressurized irrigation projects (equal
performed from 1990 to 2007 in Golestan province, to 12.45 percentage of area) are allocated to micro
Table 1- Number and area (hectare) of pressurized irrigation projects
Sprinkler systems Micro systems Pressurized systems
number % area % number % area % number area
956 93.73 20129.3 87.55 64 6.27 2861.6 12.45 1020 22990.9
224 Journal of Research in Biology (2011) 3: 223-229
Kaboosi.,2011
irrigation systems. Main reason of non-development which can cause irrigated land area increases. As
of micro irrigation in Golestan province is the little mention previously and shown in table 2, in
area of orchards in this province. Golestan province, area of orchards is much lesser
2- Percentage of agricultural lands equipped than farmland area. Therefore, in spite of little area
with pressurized irrigation systems of micro irrigation projects, its percentage is more
Area and percentage of agricultural lands than the percentage of sprinkler irrigation projects.
equipped with pressurized irrigation systems in 3- Comparing development of pressurized
Journal of Research in Biology
Golestan province is present in the table 2. irrigation in Golestan province with the country
According to this table, only 6.64 percentages of Area and number (N.) of pressurized
total irrigated agricultural lands is equipped with irrigation projects that performed in Golestan
pressurized irrigation systems that it is equal to 3.2 province within a period of 18 years are presented
percentages of total agricultural lands. It was clearly in table 3 and figure 1. In addition, area of
shown that in a period of 18 years, pressurized pressurized irrigation projects that performed in
irrigation systems have not developed in this country (Iran) after Zareii and Sadre Ghaen (2005)
province and at the present about 95% of total are shown in this table and figure. As seen,
irrigated agricultural land area is irrigating with development trend of pressurized irrigation systems
traditional surface method. Whereas more than 50 in Golestan province is completely similar to that of
percentages of total agricultural lands in Golestan the country. The most number and area of
province is rainfed, obviously water saving by the pressurized irrigation systems in Golestan province
development of pressurized irrigation systems and country are performed in 1996.
Table 2- agricultural land area (hectare) equipped with pressurized irrigation
area of
Type cultivation area of land % of total %
systems
irrigated 327248 45.6 6.15
farmland 20129.3
total 684766 95.43 2.94
irrigated 19243.4 2.68 14.87
orchards 2861.6
total 32828.6 4.57 8.72
irrigated 346491.4 48.29 6.64
total 22990.86
total 717594.6 100 3.2
Table 3- comparing development trend of pressurized irrigation in Golestan with the country
Sprinkler systems Micro systems Pressurized systems
year Golestan Iran Golestan Iran Golestan Iran
N. area area N. area area N. area area
1990 0 0 2450 0 0 350 0 0 2800
1991 2 21 8594 0 0 2406 2 21 11000
1992 4 65.5 10629 0 0 2098 4 65.6 12727
1993 35 577 10778 1 3 2695 36 580 13473
1994 74 1459.6 23476 2 1 3024 76 1460.3 26500
1995 38 684.6 9875 1 1 1725 39 685.6 11600
1996 262 4454.7 66112 1 1.6 9288 263 4456.3 75400
1997 195 3304 37058 4 43 8442 199 3346.9 45500
1998 87 2031.5 27939 1 5 11061 88 2036.5 39000
1999 57 1735.1 20308 1 48.6 11692 58 1783.7 32000
2000 69 1501.8 19293 2 151 13607 71 1652.8 32900
2001 20 287.1 14467 2 126.2 9710 22 413.3 24177
2002 9 83.2 12499 3 101 15294 12 184.2 27748
2003 23 895.3 28061 4 844.4 27138 27 1739.7 55199
2004 29 526 * 5 598.8 * 34 1123.8 *
2005 16 902.4 * 8 233.9 * 24 1136.3 *
2006 25 1391.5 * 17 559.6 * 42 1951.1 *
2007 11 208.9 * 12 144.5 * 23 353.4 *
total 956 20129.3 * 64 2861.6 * 1020 22990.9 *
* Non available data
Journal of Research in Biology (2011) 3: 223-229 225
Kaboosi.,2011
Journal of Research in Biology
Figure 1- comparing development trend of pressurized irrigation in Golestan with the country
systems area, respectively. However, sum of the soybean, wheat and barley. Others sprinkler
number and area of center pivot, linear move and projects are performed for corn and canola. Based
side roll systems in Golestan province is in 26 on published statistical information by agriculture
projects and 2510.8 hectare, respectively. This is ministrys head-office of Statistics and Information
equal to 2.72 and 12.47 percentage of total number (2005), 76 percentage of total farmlands area in
and area of sprinkler irrigation systems. The most Golestan province (more than 520000 hectare) are
important reasons of little development in center allocated to these four crops (cotton, soybean,
pivot, linear move and side roll systems in Golestan wheat and barley), in that 43 percentage of it is
province are the existence of cheap workers in the irrigated farmland. Therefore, at present only 8
region and petty landowners. Since too much percentage of irrigated land area of these four crops
percentage of agricultural lands in Golestan is equipped with sprinkler irrigation systems.
province have small area; developing of these Staple percentage of micro irrigation systems in
systems is difficult. Golestan province is point source emitter system
Hand move 2 system has the least amount of that allocated to orchards as plum, peach,
area average. Its area average is 11.5 hectare. eucalyptus, kiwi, olive and citrus trees while line
However, center pivot and linear move systems source emitter systems mostly allocated to some of
have the most area average. Their area averages are the farmland crop as cotton, tomato and soybean.
213.5 and 117.3 hectare, respectively. 6- Variety of users of pressurized irrigation
As seen in table 4, staple percentage of systems in Golestan province
micro irrigation systems in Golestan province is In this research, users of pressurized
point source emitter system that allocated to irrigation systems in Golestan province is divided to
orchards while 28.27 percentage of micro irrigation 3 groups: 1- personal or individual farms 2-
systems area is line source emitter system that Governmental farms and 3- cooperative farms.
allocated to some of the farmland crop. Table 5 shows the variety of users of pressurized
irrigation systems in Golestan province. As seen,
staple percentage of pressurized irrigation projects
Agriculture ministrys head-office of Statistics Zareii G and Sadre Ghaen SH. 2005. Perspective
and Information. 2005. Iranian Agriculture of Sprinkler Irrigation Development in Iran on 1400
Ministry, http://www.agri-jahad.org. Horizon. Proceeding of the Technical Workshop of
Sprinkler Irrigation, Iranian National Committee on
Irrigation and Drainage, Tehran, Iran.
Authors: ABSTRACT:
Murugesan S and
Muthusamy M. The Western Ghats is one among the hot spots in 25 biodiversity countries
identified in the world. The Coimbatore city is situated at the foot hills of Western
Ghats (a part of Nilgiri Biosphere Reserve) are recognized as a paradise of butterflies
due to its salubrious climate. Butterflies in the Western Ghats belong to 5 families,
Institution:
Post graduate and Research 166 genera and 330 species, of which 37 species are endemic and they depend on
Department of Zoology, more than 1000 plant species for feeding and breeding. The present study surveyed
Government Arts College 103 individual butterfly species belonging to 5 families namely Nymphalidae (32),
(Autonomous) Pieridae (23), Lycaenidae (19), Hesperiidae (15) and Papilionidae (14), which revealed
Coimbatore 641 018. that Nymphalidae and Pieridae are the rich dominant families, while Hesperiidae and
Tamil Nadu, India. Papilionidae are less dominant. High incidences of butterfly population with wide
distribution were observed during the months of March-April and the monsoon
seasons (September - November) which diminish during December-January. It was
observed that the occurrence and distribution of butterflies are closely associated
Corresponding author: with the availability of its larval and adult host plants. The butterfly population of a
Murugesan S species is gradually decreasing in number due to human interference in the habitat
and the destruction of host plants.
Email:
spmkutty@gmail.com Keywords:
Biodiversity, Bioregion, Butterfly population, Host plant, Western Ghats.
Dates:
Received: 06 Jul 2011 /Accepted: 08 Jul 2011 /Published: 20 Jul 2011
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.
14 sub families out of which 10 sub families were related to the available nutritional values of the host
recorded in Peninsular India. In the present work plant (Slanksy, 1992; Scribes, 1995 and Thomas,
there are 32 species of Nymphalidae butterflies 1995). The growth rate of Lepidopteran depends on
were identified (Table 4). The family Lycaenidae, the host plants consisting of nutritional composition
otherwise known as Blues is the worlds largest and plant secondary compositions (Salnsky, 1992).
family of butterflies, which about 6,000 species. In the Southern Nilgiris and on the lower plateau,
India has approximately 450 species, which is 21% March and September - October is the best seasons
of its total number of butterfly species. The present
for butterflies; May is comparatively poor month
studies viewed around 19 species of lycaenidaes (Wyhter & Blyth, 1957) for butterflies. The
were identified in the foothills of Western Ghats maximum number of species was observed in the
(Table 5). Family Nymphalidae throught out the entire study
area. Many members of this family were
DISCUSSION polyphagous which would help them to live in all
Climatic changes are thought to alter the habitats and in different elevation gradients
distribution and availability of animals and plants (Sreekumar & Balakrishnan, 2001). They also
through out the world (Warren et al., 2001). reported that the May month was comparatively
Feeding and reproduction of butterflies are closely poor month for butterfly population.
Evans WH. 1932. Identification of Indian Ugarte E and Rodricks. 1960. Butterflies of Palani
butterflies Bombay natural History, Bombay (TA). hills: A complementary list. Journal of the Bombay
Natural history society. 57(2):270-277.
Goankar H. 1996. Butterflies of Western Ghats,
India (including Srilanka): A biodiversity Warren M, Shill J, Kthomes JA, Asher J, Jox R,
assessment of a threatened mountain system. Report Huntlay B, Roy DB, Felter MG, Jeffcate S,
submitted to the center for Ecological Sciences, Harding P, Teffcoate G, Willis SG, Greateo Rex,
Bangalore (unpublished). Davies JN, Moss D and Thomas CD. 2001.
Nature 414:65-69.
Kunte K. 1997. Seasonal patterns in butterfly
abundance and species diversity in four tropical Wynter-Blyth. 1957. Butterflies of the Indian
habitats in northern Western Ghats. Journal of region. Bombay Natural History Society. Bombay.
biosciences 22(5):593-603. 523.
Authors: ABSTRACT:
Parthipan M, Binu
Thomas and Rajendran A.
Institution:
The present paper highlights the habitat variability, morphological features,
Department of Botany,
School of life Sciences systematic analysis and multi potentiality of Aloe barbadensis Mill. were collected
Bharathiar University, from Nilgiri district of Southern Western Ghats of Tamil Nadu, India. The epiphytic
Coimbatore 641046 nature of Aloe barbadensis Mill. is quite interesting than other habitats.
Tamil Nadu, India
Corresponding author:
Binu Thomas
Keywords:
Email: Aloe barbadensis, Habitat diversity, Southern Western Ghats, Nilgiri, Tamil
binuthomasct@gmail.com Nadu.
Dates:
Received: 24 Jul 2011 /Accepted: 25 Jul 2011 /Published: 30 Jul 2011
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.
INTRODUCTION they are not found in cold habitat regions and they
Most of the Angiosperm species shows are mostly found in tropical and subtropical regions
varied habitat specificity such as Lithophytes, of the country (Frame, 2003).
Chasmophytes, Psammophytes, Hydrophytes,
Xerophytes, Parasites and Epiphytes (Annaselvam RESULTS:-
and Parthasarthy, 2001; Das et al. 2006). Habitat diversity of Aloe barbadensis Mill
Angiosperms have been so successful in terrestrial The Aloe barbadensis Mill. is a perennial
and aquatic ecosystems that they represent majority plant with fleshy leaves. A native of North Africa.
of the herbs and shrubs and many of the trees as It is planted as hedge in house premises and also
well. Depend up on their mode of nutrion and run wild in the desert conditions and poorest soils
attachment pattern they are of different types and and also extensively cultivated throughout India for
shows variety of growth forms and also adapted to its medicinal value. The habitat variability of the
various habitats for their survival (Rao, 1994). Aloe barbadensis Mill, reveals that it is not only in
The species Aloe barbadensis Mill, comes terrestrial habitat but also in rocky habitats as
under the family liliaceae. Most of the members of chasmophytes in the Southern Western Ghats of
the family liliaceae are herbs, sometimes climbers Coimbatore district of Tamilnadu by Binu Thomas
and rarely as shrubs. The roots are fibrous, tuberous et al. (2009). The present study observed that the
or a creeping rhizome. Leaves various, cauline or species Aloe barbadensis Mill. shows an intresting
radical and sometimes functionally replaced by epiphytic habitat of Aloe barbadensis Mill. in the
cladodes, fleshy, usually parallel veined. Flowers blue mountains (2,240 meters above sea level) of
usually regular and 2-sexual, axillary or terminal Nilgiri District of Southern Western Ghats of Tamil
solitary. Fruit become berry, seeds globose or Nadu shows an intresting epiphytic habitat of Aloe
flattened. The main characteristic feature of the barbadensis Mill. It is well established in tree
species Aloe barbadensis Mill. having thick and trunks of Grevillea robusta Cunn. (Proteaceae).
fleshy leaves. The margins of the leaves become The nature of unusual/uncommon habit of Aloe
spinous and forming rosettes. It is introduced from barbadensis Mill in epiphytic habitat is luxuriant
tropical Africa and naturalized in India growing when comparing with the same species grown as
and run wild, especially in hedge- rows in the drier terrestrial habitat of neighbouring area of Nilgiri
localities up to 2,500 feet (Gamble and Fischer, district. The interactions between climate and
1915-1936). vegetation results in the marvelous vegetation in
Aloe barbadensis Mill. is a semi-tropical Nilgiri district is very important in the contribution
plant which looks more like a cactaceous member of biodiversity of the Western Ghats (Plate-1)
of lily family which usually grows in the African Systematic and Morphological analysis of Aloe
continent. There is an evidence to suggest that Aloe vera (Liliaceae).
barbadensis originated from the warm, dry climates Aloe barbadensis Mill. f.,Fl. Ind. 83.1768;
of Africa (Moon, 1824). Now the plant is much Aloe perfoliata L., Sp. Pl. 320. 1753; Gambles Fl.
more widespread and can be found growing Pres. Madras 1520. 1928; Henry et al., Fl. Tamil
throughout Europe and North America as well as Nadu 37.1989; (Kathalai).
South America, the Middle East, China, India, The dwarf plant with radical rosettes leaves,
Pakistan and Australia (Eldridge, 1978). The habitat ensiform, 40-60 2-8 cm, succulent, spiny. Scapes
of Aloe vera means Woodlands, Mediterranean 1-3; racemes to 40 cm. Flowers bisexual, yellow,
forests and Scrub according to the World Wildlife 2.5-3.5 cm long. Perianth-tube terete, 1-1.5 cm,
Federation ( Balakrishnan,1974). somewhat curved; lobes 6, orange, oblong, 1 0.5
The Aloe barbadensis Mill. habitat needs cm, 3- nerved. Stamens 3 + 3. Ovary 3-celled;
direct rays of the sun and a well drained soil. When ovules , axile; style elongate to 2.5 cm; stigma
these plants are grown outdoors then it needs obscurely lobed. Capsule ellipsoid-oblong to 1.5
warmth sun rays and protection from the winters. 1 cm.
Sometimes the habitat is destroyed due to various The Aloe barbadensis Mill. plant has thick
factors. The reasons are settlement of area by juicy leaves with sharp points, which grow to a
humans and deforestation (Wilfred and Claudia, height of twelve to sixteen inches. The leaves of
2007). It disturbs the natural balance of the area and aloe vera have no stem and are greenish in color.
destroys most of the species of Aloe vera plants. Too much water in the soil makes the leaves pale
The Aloe vera plants consist mostly of 95% water and sunlight again restores the color. Too much
238 Journal of Research in Biology (2011) 3: 237-241
Parthipan et al.,2011
Plate 1. Epiphytic nature of Aloe barbadensis Mill. on tree trunks of Grevillea robusta Cunn.
water is not good for survival of the plant. The leaf is also used for rock gardening or rockery (Binu
tissues having a gel known as aloe gel. The leaves Thomas et al., 2011).
of the plant appear to be sword type and have small Aloe barbadensis Mill. is used in alternative
harmless spines on the entire edge of the plant. The medicines and in home first aid
leaves of Aloe barbadensis Mill. is made up of four A brief account of Aloe barbadensis Mill.
layers such as Rind- the outer part of protective reported to be used in folk medicine in different
layer, Sap- a layer of bitter fluid which helps to parts of India as well as mentioned in the literature.
protect the plant from animals, Mucilage Gel- the
inner part of the leaf that is filleted out to make DISCUSSION AND CONCLUSION
Aloe gel and Inner gel- It contains 8 essential Aloe barbadensis Mill. usually found in
Amino acids (Shen et al., 2001). The root and the terrestrial habitat but the studies on the diversity
leaves both are fibrous as it holds enough water in it and distribution of chasmophytic plants from
and the leaves have dots on the entire surface. The coimbatore district of Tamil Nadu shows that the
plant bears fruit which are triangular in shape and plant like Aloe barbadensis Mill. also lives in rock
has many seeds in it crevices (Binu Thomas et al., 2009). The present
Importance of Aloe barbadensis Mill. in various study on the habitat variability of Aloe barbadensis
aspects Mill. reveals that, it also found on tree trunks. The
Using as Indoor Plant epiphytic nature of the highly medicinal plant like
Aloe barbadensis Mill needs direct sunlight for Aloe barbadensis Mill. was proved through the
growth. Some people keep it as indoor plant but the present observation. This plant also shows a
plant has to be kept in sunlight every alternate day xerophytic nature. The tissues of the plant body
(Kapoor and Sharga, 1993). stores large amount of water, thereby it can
Used for rock gardening withstand at any environmental conditions.
The xerophytic species like Aloe barbadensis Mill.
Frame G. 2003. Generalist flowers, Biodiversity Paulsen E, Korsholm L and Brandrup F. 2005. A
and Florivory Implications for Angiosperms double blind, Placebo-controlled study of a
Origins. Taxon., 52:681-685. commercial Aloe vera gel in the treatment of slight
moderate Psoriasis vuigaris. J. D e r m a t o l . a n d
Gamble JS and Fischer CEC. 1915-1936. The venereol. 19: 326-3Randhawa, G.S.1973.
Flora of Presidency of Madras. Part 1-11(Part 1 - 7
by Gamble and 8- 11 by Fischer) Adlard and Sons Rai MK. 1985, Plants used as medicine by tribals
Ltd., London. (Repr. ed. vols. 1-3.1957). of child wara District ( M . P. ) J .E co n.T ax . Bot . ,
7:385-387.
Grindly D and Reynolds T. 1986. The Aloe
barbadensis Mill.Phenomenon: A review of the Rao RR. 1994. Biodiversity in India (Floristic
properties and modern uses of the leaf parenchyma aspects) Bishen Singh Mahendra Pal Singh, Dehra
gel. J. Ethnopharmacol., 16:117-151. Dun.
Authors: ABSTRACT:
Tambekar DH, Patil RV
and Pawar AL.
Methanotrophic bacteria were isolated and characterized from sediment
from alkaline Lonar Lake. Four bacterial strains were isolated using minimal salt media
to study the methanotrophs of Lonar Lake and selected bacterial strains were further
Institution: characterized, screened on the basis of the temperature, and salt tolerance. Bacterial
Post Graduate Department of isolates were subjected to morphological, biochemical characterization and 16S rRNA
Microbiology, sequencing. Isolates were related to the phylum proteobacteria contains different
S.G.B. Amravati University, genera such as Acinetobacter baumannii, Pseudomonas aeruginosa, Achromobactor
Amravati 444602 (India). xylosoxidans and Ochromobactrum tritici. These results clearly showed that the Lonar
lake ecosystem harbors unexplored methanotrophs which can be used to control
global warming as well methanol remediation.
Corresponding author:
Tambekar DH
Keywords:
Email: Lonar Lake, methanotrophs, Acinetobacter, Pseudomonas, Achromobactor
diliptambekar@rediffmail.com and Ochromobactrum.
Article Citation:
Web Address:
Tambekar DH, Patil RV and Pawar AL.
http://jresearchbiology.com/
Documents/RA0064.pdf. Studies On Methanotrophs from Lonar Lake
Journal of research in Biology (2011) 3: 230-236.
Dates:
Received: 19 Jul 2011 /Accepted: 25 Jul 2011 /Published: 29 Jul 2011
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.
Project release 10.26, in order to identify chimeras. methanotrophs. A preliminary study was carried
Phylogenetic analyses were performed using the out to isolate the bacterial strains from sediment
ARB software package. The 16S rRNA gene using Minimal salt broth. Data of four sediment
phylogenetic analyses were performed by the sample as per the characterization are conformed up
maximum-likelihood method, using 1,285 to 1,392 to generic the level.
nucleotide positions. The functional genes were Four different isolated strains were
translated into amino acid sequences, and these identified by using standard procedures. The
were included in phylogenetic analyses using the experimental outcome of morphological and
neighbor-joining method (Dayhoff PAM model). biochemical characterization proved that all four
stains are Gram negative and identified as
RESULTS AND DISCUSSION: Acinetobacter, Pseudomonas, Achromobactor,
Methanotrophic bacteria are a group of Ochromobactrum which prove to survive in
organisms with the ability to use compounds with extremophilic condition of high salt concentration
no carbon-carbon bonds (C1 compounds) as single and high pH (10.5). Present study reported that 4
sources of carbon and energy, thus playing a role in methanotrophic gram negative, bacterial isolates
global carbon cycling. Traditional microbial from the sediments of the Lonar Lake. Among
techniques such as enrichment and isolation on four isolates Acinetobacter baumannii was oxidase
defined culture media have revealed that negative and other three Pseudomonas aeruginosa,
methanotrophic bacteria occur in a variety of Achromobacter xylosoxidans and Ochromobactrum
environments, such as freshwater, marine, and tritici were positive. Catalase, indol, methyl red,
terrestrial habitats, including habitats characterized voges-proskauer were negative and citrate
by extreme conditions of temperature, salinity, or utilization was positive in all isolates. On the basis
pH. Active members of the bacterial community in of sugar fermentation, Acinetobacter baumannii
the sediment of Lonar Lake with special emphasis fermented dextrose the other isolates can not
on C1 utilizers (Methanotrophs) were identified by ferment the sugars; nitrate reductase test was
employing two complementary culture-dependent positive for Achromobacter xylosoxidans and
and independent approaches: morphological and negative for all other. Urease production was absent
standard biochemical identification and 16S rRNA for all isolates. Among four bacterial strains
analysis probing C1 substrates methanol. The Acinetobacter baumannii showed clear zone of
objectives of this study were to isolate, identify starch hydrolysis indicate amylase production. The
methanol utilizing bacterial from Lonar Lake and present studies on optimization of various growth
study further to evaluate their potential as parameters such as temperature, salt concentration
Table 1: Morphological, cultural, biochemical characteristics and 16S rRNA identification of bacteria isolated
from sediments of Lonar Lake
and the strains showed better growth at 42C and aerugi nosa ( Lonar B) , Achromobact er
6.5% salt concentration (Table 1). All the isolates xylosoxidans (LonarC) and Ochromobactrum
were analyzed by 16S rRNA analysis and found, tritici (LonarD) (Table 2). A related work was
Acinetobacter baumannii (LonarA), Pseudomonas carried out by Jurjen Heyer (2005). It was evidently
Table 2a: 16S rRNA analysis (Blast analysis), Dendrogram showing phylogenic relationship of methanotrophs isolated
from Lonar Lake
Lonar A:
GATGACGCTGGCGGCAGGCTTAACACATGCAAGTCGAGCGGGGGAAAGGTAGCTTGCTACTGGACCTAGCGGCGGACGGGTGAGTAATGCTTAGGAAT
CTGCCTATTAGTGGGGGACAACATCTCGAAAGGGATGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGATCTTCGGACCTTGCGCTAATAGATG
AGCCTAAGTCGGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCTGTAGCGGGTCTGAGAGGATGATCCGCCACACTGGGACTGAGACA
CGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGGAACCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGCCTTATGGTTG
TAAAGCACTTTAAGCGAGGAGGAGGCTACTCTAGTTAATACCTAGGGATAGTGGACGTTACTCGCAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCC
GCGGTAATACAGAGGGTGCGAGCGTTAATCGGATTTACTGGGCGTAAAGCGTGCGTAGGCGGCTTATTAAGTCGGATGTGAAATCCCCGAGCTTAACTT
GGGAATTGCATTCGATACTGGTGAGCTAGAGTATGGGAGAGGATGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGAT
GGCGAAGGCAGCCATCTGGCCTAATACTGACGCTGAGGTACGAAAGCATGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGT
CTACTAGCCGTTGGGGCCTTTGAGGCTTTAGTGGCGCAGCTAACGCGATAAGTAGACCGCCTGGGGAGTACGGTCGCAAGACTAAAACTCAAATGAATT
GACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGCCTTGACATACTAGAAACTTTCCAGAGATGGA
TTGGTGCCTTCGGGAATCTAGATACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTTTCC
TTACTTGCCAGCATTTCGGATGGGAACTTTAAGGATACTGCCAGTGACAAACTGGAGGAAGGCGGGGACGACGTCAAGTCATCATGGCCCTTACGGCCA
GGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCTACACAGCGATGTGATGCTAATCTCAAAAAGCCGATCGTAGTCCGGATTGGAGTCTGCA
ACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGCGGATCAGAATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGG
AGTTTGTTGCACCAGAAGTAGCTAGCCTAACTGCAAAGAGGGCGGTTACCACGGTGTGGCCGATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGG
GAA
Lonar B:
TCAGATTGAACGCTGGCGGCAGGCCTAACACATGCAAGTCGAGCGGATGAAGGGAGCTTGCTCCTGGATTCAGCGGCGGACGGGTGAGTAATGCCTAGG
AATCTGCCTGGTAGTGGGGGATAACGTCCGGGAAACGGGCGCTAATACCGCATACGTCTGAGGGAGAAAGTGGGGGATCTTCGGACCTCACGCTATCAG
ATGAGCCTAGGTCGGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAG
ACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGG
ATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCAGTAAGTTAATACCTTGCTGTTTTGACGTTACCAACAGAATAAGCACCGGCTAACTTCGTGCCAGCAG
CCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTCAGCAAGTTGGATGTGAAATCCCCGGGCTCAAC
CTGGGAACTGCATCCAAAACTACTGAGCTAGAGTACGGTAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAG
TGGCGAAGGCGACCACCTGGACTGATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGAT
GTCGACTAGCCGTTGGGATCCTTGAGATCTTAGTGGCGCAGCTAACGCGATAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAA
TTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCTGGCCTTGACATGCTGAGAACTTTCCAGAGATG
GATTGGTGCCTTCGGGAACTCAGACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGT
CCTTAGTTACCAGCACCTCGGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGC
CAGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTG
CAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGG
GAGTGGGTTGCTCCAGAAGTAGCTAGTCTAACCGCAAGGGGGACGGTTACCACGGAGTGATTCATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGG
GAACCTGC
clone P5D18-571
Pseudomonas sp. DG2b
Uncultured bacterium clone P5D18-561
Uncultured bacterium clone P5D18-526
Uncultured bacterium clone P5D15-458
Uncultured bacterium clone P5D4-567
Uncultured bacterium clone P2D11-424
Uncultured bacterium clone P2D11-425
Uncultured bacterium clone P2D11
Pseudomonas aeruginosa DQ989018
Pseudomonas aeruginosa Lonar-B
Table 2b: 16S rRNA analysis (Blast analysis), Dendrogram showing phylogenic relationship of methanotrophs
isolated from Lonar Lake
Lonar C
TCAGATGAACGCTAGCGGGATGCTTACACATGCAAGTCGAGCGCAGCACGGACTTCGGTCTGGTGGCGAGTGGCGAACGGGTGAGTAATGTATCGGAAC
GTGCCCAGTAGCGGGGGATAACTACGCGAAAGCGTAGCTAATACCGCATACGCCCTACGGGGGAAAGCAGGGGATCGCAAGACCTTGCACTATTGGAG
CGGCCGATATCGGATTAGCTAGTTGGTGGGGTAACGGCTCACCAAGGCGACGATCCGTAGCTGGTTTGAGAGGACGACCAGCCACACTGGGACTGAGAC
ACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGGGAAACCCTGATCCAGCCATCCCGCGTGTGCGATGAAGGCCTTCGGGTT
GTAAAGCACTTTTGGCAGGAAAGAAACGTCGCGGGTTAATACCTCGCGAAACTGACGGTACCTGCAGAATAAGCACCGGCTAACTACGTGCCAGCAGCC
GCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTCGGAAAGAAAGATGTGAAATCCCAGAGCTTAACTT
TGGAACTGCATTTTTAACTACCGGGCTAGAGTGTGTCAGAGGGAGGTGGAATTCCGCGTGTAGCAGTGAAATGCGTAGATATGCGGAGGAACACCGATG
GCGAAGGCAGCCTCCTGGGATAACACTGACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTC
AACTAGCTGTTGGGGCCTTCGGGCCTTGGTAGCGCAGCTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGA
CGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACATGTCTGGAATGCCGAAGAGATTTGGC
AGTGCTCGCAAGAGAACCGGAACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTC
ATTAGTTGCTACGAAAGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTATGGGTAGGGCT
TCACACGTCATACAATGGTCGGGACAGAGGGTCGCCAACCCGCGAGGGGGAGCCAATCCCAGAAACCCGATCGTAGTCCGGATCGCAGTCTGCAACTCG
ACTGCGTGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTGG
GTTTTACCAGAAGTAGTTAGCCTAACCGCAAGGGGGGCGATTACCACGGTAGGATTCATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTATCGGAAGG
TGCGGCTGGATCACCTCCTT
Achromobacter xylosoxidans(2)
Achromobacter xylosoxidans FM999735
Uncultured Achromobacter sp.(7)
Uncultured Achromobacter sp.(6)
Uncultured Achromobacter sp.(5)
Uncultured Achromobacter sp.(4)
Uncultured Achromobacter sp.(3)
Uncultured Achromobacter sp.(2)
Uncultured Achromobacter sp.
Achromobacter xylosoxidans
Achromobacter xylosoxidans Lonar- C
Lonar D:
AACGAACGCTGCGGCAGGCTTAACACATGCAAGTCGAGCGCCCTTTTCGGAGGGAGCGGCAGACGGGTGAGTAACGCGTGGGAACGTACCTTTTGCTAC
GGAATAACTCAGGGAAACTTGTGCTAATACGTATGTGCCCCCCTTTAAAATTCCAAGAAACATAAAATGCCTTGGCATTTTATGGGGGGGAAAGATTTAT
CGGCAAAGGATCGGCCCGCGTTGGATTAGCTAGTTGGTGGGGTAAAGGCTCACCAAGGCGACGATCCATAGCTGGTCTGAGAGGATGATCAGCCACACT
GGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGCAAGCCTGATCCAGCCATGCCGCGTGAGTGATGAA
GGCCCTAGGGTTGTAAAGCTCTTTCACCGGTGAAGATAATGACGGTAACCGGAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAA
GGGGGCTAGCGTTGTTCGGATTTACTGGGCGTAAAGCGCACGTAGGCGGACTTTTAAGTCAGGGGTGAAATCCCGGGGCTCAACCCCGGAACTGCCTTT
GATACTGGAAGTCTTGAGTATGGTAGAGGTGAGTGGAATTCCGAGTGTAGAGGTGAAATTCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTC
ACTGGACCATTACTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATGTTAGCCGTTGG
GGAGTTTACTCTTCGGTGGCGCAGCTAACGCATTAAACATTCCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCAC
AAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGCCCTTGACATCCCGATCGCGGTTAGTGGAGACACTATCCTTCAGTTCGG
CTGGATCGGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCTTAGTTGCCAGC
ATTGAGTTGGGCACTCTAAGGGGACTGCCGGTGATAAGCCGAGAGGAAGGTGGGGATGACGTCAAGTCCTCATGGCCCTTACGGGCTGGGCTACACACG
TGCTACAATGGTGGTGACAGTGGGCAGCGAGCACGCGAGTGTGAGCTAATCTCCAAAAGCCATCTCAGTTCGGATTGCACTCTGCAACTCGAGTGCATG
AAGTTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTTTTACCC
GAAGGCGCTGTGCTAACCGCAAGGAGGCAGGCGACCACGGTAGGGTCAGCGACTGGGGTGAAGTCGTAACAAGGTAGC
found that methanotrophic bacteria grow at 300C different seasons from saline and hyperalkaline
and 8.7% salt concentrations. Surakasi et al, (2010) Lonar Lake using 16S rRNA gene library analysis
also reported the phylogenFetic diversity of and demonstrated the presence of Arthrospira
bacterial communities in microbial mats of two (Cyanobacteria), Fusibacter (LAI-1 and LAI-59)
and Tindallia magadiensis (LAI-27) in post- Heyer J, Berger U, Hardt M and Dunfield PF.
monsoon and Planococcus rifietensis (LAII-67), 2005. Methylohalobius crimeensis gen. nov., sp.
Bordetella hinzii (LAII-2) and Methylobacterium nov., a moderately halophilic, methanotrophic
variabile (LAII-25) in pre-monsoon. They claimed bacterium isolated from hypersaline lakes of
the first time detection of these putative Crimea. Int J syst Evol Microbial., 1817-1826.
methanotrophs in surface mats of Lonar Lake
Methanol was primarily assimilated by Jhingran AG and Rao KV. 1954. Lonar Lake and
Acinetobacter baumannii, Pseudomonas its salinity. Geol Surv India 85:313-334.
aeruginosa, Achromobacter xylosoxidans and Malu RA, Dhabhade DS and Kodarkar MS.
Ochromobactrum tritici species from the family 2002. Diversity of Lonar Lake. J Aquat Bio., 15:16-
Methylophilaceae isolated from Lonar Lake. The 18.
majority of known aerobic methane-oxidizing
bacteria are members of either Molly C, Redmond DL, Valentine and Alex LS.
Gammaproteobacteria (type I) or 2010. Identification of Novel Methane-, Ethane-and
Alphaproteobacteria (type II), though several Propane-Oxidizing Bacteria at Marine Hydrocarbon
strains of highly acidophilic methanotrophic Seeps by Stable Isotope Probing. Appl Environ
Verrucomicrobia have also been recently isolated. Microbiol. 76(19):64126422.
Most methanotrophs are capable of growth only on
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xylosoxidans and Ochromobactrum tritici species, variability over past 11.5 kyr: A multi proxy study
are new species and not previously recorded of Lonar impact Crater Lake core, India.
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warming which may found better choice for further Surakasi VP, Antony CP, Sharma, Spatula MS
studies like methane, methanol or toxic chemical and Shouche YS. 2010. Temporal bacterial
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