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Introduction

Shrimp farming is a major industry in the tropics and subtropics. In traditional


extensive system, there follows frequent exchange of water to maintain the desirable
quality of the culture environment (Wang, 1990, Boyd and Musiri, 1993, Hopkins et al,
1999). However there is a concern on impact of aquaculture effluent on the coastal waters
which might result the leads to its deterioration (Eng et al, 1989). The nutrient rich
effluent discharged from shrimp farms can cause eutrophication of coastal waters and its
impact has been a major environmental concern (Philips et al, 1993). Previous reports on
nutrient budget revealed that in such open shrimp culture system as much as 90% of
nitrogen and phosphorus input is in the form of feed, out of which the major portion is
lost to the system with only less than one sixth being assimilated as the shrimp biomass
(Muthuvan, 1991, Briggs and Funge – Smith, 1994).
Another major issue faced by shrimp industry is the diseases caused by potential
and opportunistic pathogens. Among them the most serious out breaks were of viral in
origin triggered by poor water quality and the situation sharing the same source water
(Burford et al 2003). In this scenario measures to prevent the introduction of pathogen
have become a necessity to protect the crop from onslaught of disease as well as to
prevent the discharge of waste water in to the culture environment as a long term strategy.
It has been pointed out that at present there are no therapeutic measures available for
shrimp viral disease. However several farm management strategies have been developed
to prevent viral entry in to the culture system and subsequent disease out breaks. As a
result of the pressure faced by the shrimp farming community for increased biosecurity,
diseases and effluent control (Bullis and Pruder, 1999), there has been a trend within
many countries towards the development of biosecure closed shrimp production system,
including zero water exchange or recirculating culture system (McIntosh, 1999,
Avnimelech, 2000). The major problem associated with closed system is rapid
eutrophication in ponds and phytoplankton bloom crash common and re blooming
comparatively fast, there is still a period of stress to the shrimp due to the nutrients and
organic matter enrichment over the culture period. The super eutrophic pond water can
decrease the carrying capacity of the pond (Lin, 1995). After 120 days stressfull condition
develops and growth rates are slower compared to open system. Obviously the balance
between waste production and assimilative capacity in pond environment is of paramount
importance for the success of closed system.
Pond bottom conditions are more critical for shrimp than for other aquaculture
species, because shrimp spend most of their time on the bottom soil (Boyd, 1989).The
distribution of penaeid shrimp in the natural environment can be influenced by sediment
characteristics (Williams, 1958).In intensive shrimp culture system the presence of
sediment and other surface areas may have both positive and negative effects on shrimp
production ( Bratwold et al 2001). From a chemical stand point, particularly in closed
system with minimal or no water exchange sediment can be a source or sink for
macronutrients such as Cu,Fe,Mn and Zn than those found in the water column
(Horowitz, 1999). Aquaculture pond bottom soils are recipient of large amount of
nitrogen, phosphorus and organic matter (Boyd, 1990, 1992 ) and these substances tend
to accumulate in bottom soils. High organic matter in bottom soil may result in anaerobic
conditions in the surface layer of soil and soil water interface. When oxygen is depleted,
other electron acceptors can be used to mediate the decomposition of organic matter.
Many anaerobic processes taking place in the pond bottom lead to the production of
reduced and potentially toxic compounds like sulphide and methane (Boyd, 1992). Under
such situations use of probiotics in aquaculture settings has been increasing with demand
for more environment friendly aquaculture practices (gatesoupe, 1999). In present years,
the biological control of disease by environmental friendly methods such as probiotics
has become an important subject of investigation in aquaculture research. (Mathew
castex, 2008, Garriquez and Arevalo 1995, Gomezgil et al 2001, Vine etal 2006) detail
the various developments made in the use of probiotics in aquatic cultured species
including shrimp. The main probiotic bacterium documented in shrimp grow out are
Bacillus species strains (Moriarty 1998, Ziaed nejad 2006) or gram negative bacterial
strains (Alavandi et al , 2004, Vijayan et al 2006) . Many works have evaluates some
special microorganisms biologically improves water quality. Douilett (1998) used a
probiotic addition consisting of a blend of bacteria in a liquid suspension in intensive
production system. The probiotic blend improved water quality in fish and crustaceans
culture by reducing the concentration of organic materials and ammonia. This procedure
was accomplished by a series of enzymatic processes carried out by the organism.
National Centre for Aquatic Animal Health developed a bioaugmentor ‘Detrodigest’
composed of an indigenous bacterium Bacillus MCCB 101 for the management of
detritus generated during the culture period. The organism rapidly digest the detritus into
monomeric substances and then in to CO2 and water. A gut probiotic ‘Enteotrophotic’ is a
combination of two organisms such as Bacillus MCCB 101 and Arthrobacter MCCB
104. The Arthrobacter MCCB 104 is a marine isolate with extra cellular antagonistic
property against several species of Vibrio such as V. alginolyticus, V. parahaemolyticus,
V. vulnificus, V. neris and Aeromonas species.
Using these probiotic preparations a zero exchange shrimp culture practice was
developed by the National centre for Aquatic Animal Health. The study was conducted to
evaluate and validate the systems in terms of environmental quality and productivity.
Materials and Methods.
Bioassay study
This study aims to evaluate the influence of Detrodigest in improving the Eh of
soil. It was carried out in 100 L capacity fiber glass (two control and two test tanks). All
the tanks were provided as uniform 20 cm layer of sediment taken from a shrimp culture
pond. The bioaugmentor of 5 ml aliquots directly added to the test tanks seven days
intervals continuously and the water and soil parameters measured regularly.
Field study
Location and description of the pond
Ten ponds of 1 hector and two 1.5 hector ponds were selected at Pancham
Aquaculture farms Ltd, Mumbai, Maharastra, India. A set of eleven ponds was used as
test and one as control. Pond preparation and management practices followed during the
culture period are given below.

Pond Preparation

Five sampling stations are fixed in a pond, and 100g soil samples taken from each
pond and divided into two parts. One part is separated for Eh and pH and the other
component from all the sampling stations are mixed together, dried, packed separately for
total soil analysis. Eh and pH are measured immediately and other analysis subsequently.
The ponds were eradicated by applying lime (1275 Kg/ha) and ammonium sulphate (at
rate of 5:1(255 kg/ ha) and tea seed powder at the rate of 100 kg/ha after maintaing the
water level to 10 cm. ( Jhingaran, 1988, Pillai, 1995). On completing the eradication the
probiotic ‘Detrodigest’ apply to effect speedy degradation of the dead weed animals.
Water is pumped in to the pond to have maximum 5-10cm at the top most portion of the
pond through 100 micron bag net fitted to the sluice gate. The sluice gate then closed by
mud packing. Recorded salinity and pH of the water taken in.‘Detrodigest’ is brewed in
creek water at a rate of 300ml/ 100 L / hectare and applied on the pond. Periodically
(once in 24 hours) pH and Eh of soil are measured in all the five stations and examined
the impact of the treatment in improving Eh.The process (application of Detrodigest) is
continued till the TOC dropped to 2% and Eh rose to around -100 mV.Fresh water is
added from reservoir to bring salinity of the disinfected water to 15 ppt. Care is taken to
replenish evaporation loss by adding fresh water through out the culture.

Fertilization

Water sample collected from the ponds are subjected for complete set of analysis
to decide the extent of fertilizers required. Inorganic fertilizers are supplemented as per
requirements to initiate bloom. A combination of nutrients such as nutrimix 650 gm
(NCAAH, CUSAT), triple super phosphate 15 Kg/ha (FACT, Kerala) and 100L cow dung
juice, were applied to the ponds during morning hours after regulating pH to 7 – 7.5 by
applying dolomite. Nutrimix per Kg contains Sodium nitrate (588g), Sodium phosphate
(39 g), Sodium silicate (270 gm), Copper Sulphate (25g), and ferric chloride (78g). In
addition another nutrient preparation ‘ micromix’ containing ferric chloride and
magnesium chloride at a ratio of 1:1 was also applied at a rate of 300 ml/ ha. Under the
situation were delay the phytoplankton failed to appear with in three days of application,
an additional dosage of 40 L extract from 40 kg cow dung, 200 g nutrimix, 620 g single
super phosphate, and 200 ml micromix were applied /hector this stage one more
comprehensive analysis of water used to be made to asses the suitability of ponds for
stocking. In the control sets of ponds, no treatment is given, but all analysis made as in
the experimental ponds.
Stocking and management of the ponds

When the water and sediment pH remained between 7- 8.5, sediment Eh between
-100 to -125, alkalinity between 70 to 120 ppm in the experimental ponds both
experimental and control ponds are stocked with Penaeus monodon seed, tested nested
PCR negative and qualified for stocking with regard to all other health parmeters.To
document survival of post larvae subsequent to stocking 100 of them were maintained in
a nylon hapa and counted after 48 hours perusing it to reflect the over all survival of the
pond. In cases alkalinity happens to be less than 70 ppm in the experimental ponds it used
to be elevated by application of dolomite or Ca CO3 in small doses. Hence forth regular
monitoring and management of the system carried out. The probiotic Detrodigest applied
in treated ponds regularly seven days interval .All physical and chemical parameters of
water and soil measured regularly. As the bloom develops readily it used to be not
essential to go for applying inorganic fertilizers. After one month of culture take 100
random animal samples from different part of the pond using cast net and assess the body
weight and examine critical health parameters. Feeding is regulated according the body
weight. At this stage Enteotrophotic application commences by coating on to the diet at a
ratio of 50 ml/ 10,000 animals in all meals. . In both experimental and control ponds
salinity and water level are adjusted by supplementing with fresh water.

Statistical analysis

The mean and standard deviation of the sediment and water quality parameters for each
pond were calculated. Single way analysis of variance (ANOVA) was done to the assess
the variation in different ponds. Productivity of each ponds in terms of yield per hector
was calculated and multiple regression was carried out using SPSS ( Statistical package
for the Social Science ,SPSS Inc,Chicago) for delineating the relation between water and
sediment quality parameters and yield. The average feed conversion ratio (FCR) and
Survival rates were also calculated.
Probiotics.

Detrodigest
The organism used for the preparation of Detrodigest is Bacillus cereus sensu lato MCCB
101 (Genbank Acc. No. EF 062509) isolated from aquaculture fields of Kerala and
subjected to rigorous screening procedures. Extensive field level demonstration and
validation over a period of five years has been made in Kerala and Tamil Nadu to
ascertain its suitability in shrimp culture systems and it has proved to be an appropriate
preparation for prolonged and safe detritus management in any aquaculture systems. An
extensive study on the salinity preference of the organism in Detrodigest revealed its
euryhaline nature by growing and adequately producing hydrolytic enzymes at all
salinities tested ranging from fresh water to seawater (0 - 45 ppt). The organism in
Detrodigest is highly versatile with the potential to produce a variety of enzymes such as
protease, lipase, chitinase etc. Another outstanding property the organism is its capability
to reduce ammonia, which gets liberated as a result of deamination and excretion by
shrimp. This was very much reported during the field level demonstration programme as
the ammonia content in all ponds, which received Detrodigest, was below toxic level
(<0.01ppm NH3). Rapid degradation of detritus as soon as formed by Detrodigest makes
more dissolved oxygen available at the pond bottom. As a result of the digestion of
detritus, mineralization proceeds at faster rate resulting in phytoplankton bloom,
generally three days after the addition of Detrodigest. The bloom may remain for 10 days
and for perpetual blooming repeated addition once in 10 days has been found to be
fruitful.
This is applied to the ponds by brewing (an aliquot of 300 ml preparation/1
hector) for 24-48 hours in 100 L medium composed of filtered (through muslin cloth of
100 mesh size), chlorinated (7%chlorine as sodium hypochlorite) and dechlorinated (after
6 hours by adding 20 g sodium thiosulpahte 100L) pond water from the respective pond
supplemented with cooked 100g each shrimp pond and rice bran. Generation of
fermented smell amd froth is the signs of brewing. For best results the preparation has to
be aerated from an air pump, passed through a bacterial filter of 45 µm or is manually
agitated intermittently to get final cell count of 1X 109 CFU ml. The preparation is
subsequently diluted with pond water and sprayed over the ponds during morning hours.
Enterotrophotic
Enterotrophotic is a scientific blend of Bacillus cereus sensu lato (MCCB 101) (Genbank
accession. no. EF 062509 and Arthrobacter nicotianae (Genbank accession no.
EU402968). (MCCB 104) isolated from marine environment and characterized for
application as gut probiotic. They control Vibrio in shrimp/prawn intestine by way
antagonistic activity mediated by an extracellular anti-vibrio molecule and enhances feed
acceptance and digestion by way of producing hydrolytic enzymes.

Water and sediment quality


Environmental quality of the culture ponds was determined based on few critical
physical, chemical and biological factors. The water samples were collected
between
7.00 to 8.00 a.m. using sterile plastic bottles usually from 10-15 cm
depth from the water surface. For the analysis of dissolved oxygen,
water samples were collected by BOD bottles of 300 ml capacity The
Manganese sulphate and the alkali iodide–azide reagent was added
immediately at the collection site to fix the samples for studying
dissolved oxygen. The measurement was carried out in the laboratory.
The water temperature and pH were measured using digitrel pH meter
at the time of sampling.pH and temperature measured morning and
evening and Dissolved oxygen twice in daily at the end of the culture
it’s measured four times. All other chemical analysis of water
measured weekly and biological analysis ten days interval. In the case
of soil analysis pH and eh measured weekly and all other parameters
once in month. All other analysis done the following procedures. Total
alkalinity ( titration method ), Total Ammonia nitrogen ( Phenate method ),nitrite nitrogen
(Colorimetric method ), Nitrate nitrogen ( Phenate method ), inorganic phosphate
( Ascorbic acid method), chlorophyll (spectroscopic method ), total phosphorus
( persulphate digesion method), silicate ( Molybdo silicate method ) primary productivity
( light and dark method ) of water and soil Eh, ( Eutech Eh meter ) and total organic
carbon ( ), total organic matter

Results

Bioassay study

The influence of Detrodigest on soil Eh improvement presented in fig. 1. There is


significant difference between soil Eh in test and control tanks ( P <.0002).
Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Column 1 15 -2904.5 -193.633 730.6486
-3455.2
Column 2 15 5 -230.35 404.0458

ANOVA
Source of
Variation SS df MS F P-value F crit
Between
Groups 10110.85 1 10110.85 17.82128 0.000231 4.195982
Within Groups 15885.72 28 567.3472

Total 25996.57 29
Field Study

Soil and water quality parameters measured during the culture period are given in
Table -1. On analyzing the data significance difference between the test and control
ponds in certain critical parameters were observed. They were the lower content of
nutrients such as total ammonia nitrogen, nitrite nitrogen, nitrate nitrogen and phosphate
and higher chlorophyll content. All other parameters are more or less uniform between
the two sets. However soil Eh in the control ponds fluctuate widely compared to that of
the test ponds. Details of production obtained under the above culture conditions are
summarized in Table-2. Accordingly significantly higher FCR, survival and biomass
production could be obtained. Mean while the day of culture was comparatively lower
(179 days against 189) in the test ponds even though the growth obtained was the same in
both the systems (0.20g/day).

Discussion

There is no significance differences between pH in control and treated ponds. Shrimp


culture system the optimum range of pH between 7.5 – 9 ( Tsai, 1989 ). The most ideal
pH reported for shrimp farms in the tropics ranged from 7.5 to 8.5 (Chien, 1992)..
Evening pH slightly greater the morning is due to the changes in the rate of
Photosynthesis by phytoplankton and other aquatic plants in response to the daily
photoperiod. These values agree with the studies of Thakur et al 2003 and Cowan
1999).Alkalinity of pond water is no significant difference between control and test ponds
this is because of the addition of dolomite for maintaining alkalinity in culture ponds. The
Dissolved Oxygen level of the treated and control ponds is also no significant difference
throughout the culture period DO maintained above 4 ppm except few days which was
use of paddle wheel aerators. The slightly higher level in treated ponds could be
attributed to the beneficial effect of probiotics through way of more phytoplanktonic
production and increased photosynthetic activity. These results agree with the studies of
Matis et al 2002 and Thakur et al 2003. Hopkins (1994), Hopkins and Sandifer (1996)
have successfully farmed shrimp in ponds with reduced or no water exchange, and
appropriate aeration level.

Nitrogen plays an important role in the limited discharge aquaculture system due
to its dual role, as a nutrient and toxicant (Burford and Lorenzen, 2004). Nitrogen is the
form of ammonia (NH3) and nitrite is highly toxic to shrimp, however the toxicity
depends on various factors including species tolerance, water characteristics. (Eg- pH,
temperature, salinity, dissolved Oxygen) and exposure duration (Hargreaves, 1998,
Barajas et al 2006).Mean of the TAN concentration was significantly lower in treated
ponds compared to control. It fluctuates largely in control ponds and once reached to 0.21
ppm.Burford et al (2003) reported in zero exchange systems of Belize Aquaculture ltd,
USA with plastic lining and high aeration rates perpetual high nitrite, ammonia and
phosphate concentrations. Moreover it was reported earlier that P. monodon grow out
system, even with frequent water exchange ammonia may increase up to 6.5 mg/L. (Chen
and To, 1991).TAN concentration never exceed the safe range during the culture shrimp
biomass very much below the carrying capacity of the system, and the deleterious
nitrogenous waste effectively removed by phytoplankton and microbial activity. . This
value lower than the study of Thakur et al 2003. Prabu et al (1999) and Rao et al (2000)
reported NH3-N above 1.0 ppm in shrimp culture ponds and Abraham et al ( 2004 )
observed that the mean NH4- N increased It is also considered to be a toxic metabolites
for shrimp ( Gonzalox- Felix et al 2004).Nitrite accumulation is a typical cycling of
nitrogenous compounds that occurs as a result of the establishment of nitrifying bacteria
that carryout the oxidation of ammonia ( NH4) to nitrite ( NO2 ) and nitrate ( NO3)
reported previously ( Decamp et al 2003 ).Nitrate, unlike ammonia and nitrite is less toxic
to shrimp however, high concentration ( 100 mg/L) was reported to be lethal to shrimp
(Muier et al 1991, Rijin et al 2006). Wickins (1976) also reported no adverse effect on
growth of Penaeus monodon juveniles (1.5 g) when exposed to nitrate concentration over
200 mg/L for 5 weeks. Mean nitrate concentration in treated ponds lower than control
pond. Mean value of inorganic phosphate in treated ponds significantly lower than the
control pond.This result is contrast with the findings of Boyd et al (1990 ) who mentioned
that dissolved orthophosphate concentration are usually not greater than 5-20 µg/L and
seldom exceed 100 µg/L even in highly eutrophic water. Hopkins et al ( 1993) while
working with out door lined ponds with soil on the bottom and operated with limited
water exchange, documented significantly higher reactive phosphorus levels in these
ponds than those operated with frequent water exchange. In the present study water
nutrient concentration linearly correlated with the cumulative feed input. A closed system
nutrient kept on accumulating with in the system overtime, this might be an advantage of
the closed system as the high nitrogen and phosphorus accumulated with in the system
could support the growth of natural food organism contributing ultimately to the shrimp
growth.( Thakur et al 2003).This result agree with the study of Burford et al (2003) in a
closed system phosphate concentration between 0.07 – 1.17 ppm. Aquaculture ponds
receiving feeds often have total phosphorus concentration above 0.5 ppm and total
nitrogen concentration 2 or 3 mg/L ( Boyd and Tucker et al 1998 ).The value of total
phosphorus concentration agree with the studies of Burford et al 2003 and Matias et al
2002 and it lower than the studies of Takur et al 2003 and Shariff et al 2001.. In an
experimental study on the nutrient budget and water quality in closed system shrimp
ponds by Thakur and Lin ( 2003), the total nitrogen and total phosphorus concentrations
in water and were found increased with the progress of rearing. Silicate is a major
constituent of diatoms.There is no significant difference between treated and
experimental ponds. This value is higher than the study of Prabhu et al 2001.

In the case of primary productivity there is no significant differences between treated and
control ponds. It is a classical pathway to remove mineral nutrients from water and helps
pond digestion processes and lead to reduction of mineral nitrogen pollutants in the
discharge (Hopkins et al 1995 ).This result is lower than the studies of Hepher and Prajes
(1981) and Ali (1986). Chlorophyll content in experimental ponds was significantly
higher than the test ponds. These values is higher than the studies of Cowan et al at lower
intensity ponds and Guereel et al 1999 in dry season studies.

The redox potential values and organic matter content between treated and control ponds
no significant difference but the fluctuation of Eh in control ponds through out the culture
period is more compared to treated ponds.Redox potential as low as -200mV reported in
fish farm sediments. Production of toxic gases like methane and H2S that adversely affect
the benthic organism that also be reported (Pawar et al 2002). It is possible to evaluate the
intensity of the pond anaerobic conditions by measuring the sediment redox potential
(Hutchison 1957). The Eh value of control ponds also good. This is because there is not
much problems in control ponds till the end of the culture. But in the case of survival
and FCR control ponds showed very much difference between treated ones because the
addition of probiotics improved the health condition and environment of the culture
species.

Survival and FCR

The average FCR of treated ponds 1.9 and well with in the country estimate for India
( 2.0) an equal to other countries like Indonesia, lower that of China ( 2.3 ) and Vietnam
( 2.5 ) ( FAO 2002 ). The average survival rate in treated ponds 69.5, substantially higher
compared to 26.5% survival of Chilka lagoon (Balasubramanian et al 2004). Venket et al
(2004) showed that strains of L. acidophilus and L. sporogenes significantly improved
growth of M. rosenbergii post larvae. However survival was not effected in this case. The
same authors observed inhibitory effects of the LAB s tested against the gram negative
flora present in the fresh water of the shrimp gut. The positive effects of probiotics in
FCR have also been already pointed out in P. indicus larvae feed with L.plantarum (Uma
et al 1999). In the study of Mathiew castex (2008) probiotic treatment improved final
biomass. McIntosh (2002) reported that the dietary crude protein level could be decreased
from 31% to 24% with FCR decreasing from 2.2 to 2.0 and body N retention efficiency
increased from 23 % to 37 % in shrimp reared under commercial zero water exchange
culture conditions. The average production for the reported 100,00 shrimp farms in India
with an area of 130,000 ha in production is at the rate of 538 kg ha ( Rosenberry, 1999).
Statistical Analysis
Pond management studies usually limited due to the non availability of culture systems
for experimentation to maintain adequate replicates ( Lemonnier abd Faninoz,2006) . In
such situations regression analysis of data from trials with single or duplicate ponds for
each treatment over a wide range of parameters is a more appropriate means of obtaining
Information on pond management ( Wudistin and Boyd 2005). In our study multiple
regression analysis was performed for all the ponds over the culture period. The fitted
regression equation is Growth = -80.673+- 4.820pH morn +19.290 pH Even – 21.341
NH3 – 23.850 NO2 + 28.573 NO3 +3.530 PO4 - .527 DO. The predicted regression
equation is significant ( P<.001 ) and explain 67.2 % variability in the data.

Table -1. Mean and Standard deviation of water and soil quality parameters between
treatment and control ponds.

Parameters Treated ponds Control pond


Mean SD Mean SD
pH( Morn ) 8.16 0.27 8.52 0.24
pH ( Eve) 8.57 0.28 8.52 0.24
Dissolved Oxygen ( ppm) 5.29 0.73 5.06 0.97
Alkalinity ( ppm) 70.23 9.67 71.37 10.8
TAN (ppm ) 0.01a 0.013 0.049a 0.06
Nitrite ( ppm ) 0.018a 0.007 0.04 a
0.03
Nitrate ( ppm ) 0.075 a 0.031 0.25 a
0.22
Phosphate ( ppm ) 0.099a 0.02 0.161a 0.07
Total Phosphorus (ppm ) 0.2 0.04 0.24 0.09
Silicate ( ppm) 0.08 0.16 0.07 0.02
Primary productivity 1.06 0.38 0.93 0.53
g/C/m3/day
Chlorophyll (µg/L 21.21a 6.49 15.59a 9.35
Soil Eh (mV ) -152.68 16.42 -159.4 60.14
Total organic carbon (%) 0.42a 0.11 0.3a 0.04
Same letters represents significantly different

Table- 2. Production Parameters of Penaeus monodon culture

Treated Ponds Control Pond


Day of culture 179 189
Growth rate ( g/day ) 0.20 0.20
FCR 1.9a 2.7a
Survival 69.5a 54 a
Production Kg/ha 1820.9a 1600 a
Ammonia and Nitrite level of the treated and control ponds through out the culture period

Am m onia Nitrite
0.12
0.25
0.1
0.2
0.08
Tes

NO2
0.15 Test 0.06
NH3

t
0.1 control 0.04 con
0.05 0.02 trol
0 0
1 3 5 7 9 11 13 15 1 3 5 7 9 11 13 15
w eeks w eeks

FCR and Survival of the treated and control ponds

FCR and Survival

90 3
80
2.5
70
60 2
50 Survival
survival

FCR

1.5 FCR
40
30 1
20
0.5
10
0 0
1 2 3 4 5 6 7 8 9 10 11 12
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