1. PROPOSAL FOR M.F.Sc RESEARCH PLAN
TITLE OF PROPOSED RESEARCH
Effect of different stocking density on growth and
physiological function of Ompok bimaculatus (Pabda) reared
in a biofloc system
PRESENTED BY-
SNIGDHA SUCHARITA MAJHI
ADMISSION NO : AQC- 01/19
DEPARTMENT OF AQUACULTURE
COLLEGE OF FISHERIES, CAU(I), LEMBUCHERRA, TRIPURA (W)
2. ADVISORY COMMITTEE
Chairperson
DR. SOIBAM KHOGEN SINGH
Assistant Professor , Dept. of Aquaculture
MEMBERS
PROF. ARUN BHAI PATEL
Professor & Head
Dept. of Aquaculture
DR. GUSHEINZED WAIKHOM
Associate Professor
Dept. of Aquaculture
DR. DIBYENDU KAMILYA
Assistant Professor
Dept. of Aquatic Health and Environment
DR. PRASENJIT PAL
Assistant Professor
Dept. of Extension & Social Science
5. INTRODUCTION
BIOFLOC
FCR found to be
minimum in the
presence of biofloc
(1.2- 1.29) (Khanjani,
2015)
With a nutritional
value of protein-
38%, lipid- 3%, it is
readily accepted
among various fish
species (Azim and
Little, 2008).
Gather certain
storage compounds
like PHB which
protect animals from
bacterial infections
(De schryver, et al.,
2010) Certain bioactive
properties are
imparted by bio-floc
(Khanjani and
Sharifinia, 2019).
Low investment and
high return system
6. SPECIES reported UNDERBIOFLOC system
Sl. No. Species Reference
1. (Tilapia) De Schryver et al. (2008)
2. Litopeneaus vannamei (White leg shrimp) Crab (2010)
Martins et al. (2019)
Porchas et al. (2019)
Panigrahi et al. (2020)
3. Labeo rohita (Rohu) Mahanand et al.(2013)
4. Clarias gariepinus (African catfish) Bakar et al. (2015)
Chen et al. (2019)
5. (Nile tilapia) Nahar et al. (2015)
6. Peneaus indicus (Indian White Shrimp) Panigrahi et al. (2020)
7. L. macrochirus Fischer et al. (2020)
7. PROBLEM STATEMENT AND RESEARCH GAP
BIOFLOC
RESEARCH
Limited species C/N ratio; micro-biome; Carbon sources Stocking density?
Highly complicated system
Species diversification
(local suitability/demand)
Performance vary with
species
-growth and
survivability
-economic
sustainability
-physiological
changes
8. Broad OBJECTIVES ofthe study
- To evaluate the growth, survival and water quality changes in biofloc system
for culture of O. bimaculatus at different stocking densities.
- To examine the health and related physiological attributes of O. bimaculatus
reared in biofloc system at different stocking densities.
9. NULL HYPOTHESIS
• H0: There is no influence of stocking density on growth, survival and
physiological status of O. bimaculatus reared in biofloc system.
11. STATUSIN INDIA
•Sl.no. Topic Reference
1. Influence of different protein levels of feed in L.
vannamei cultured in BFT
Panigrahi et al. (2019)
2. Biofloc with different C/N ratio in inland saline
water
Kumar et al. (2019)
3. Comparative efficiency of biofloc and feed based
culture of common carp
Sarkar et al. (2019)
4. Culture of IMC in BFT Deb et al. (2020)
5. Survival of L. vannamei from AHPND in BFT Kumar et al. (2020)
12. CATFISH experimented UNDER BFT
Sl No. Species Reference
1. I. punctatus Schrader et al.,(2011)
2. R. quelen Poli et al.,(2015)
3. C. gariepinus Ekasari et al., (2016)
4. P. hypothalamus Hoa et al., (2017)
5. C. gariepinus Dauda et al., (2018)
6. Silurus asotus Lee et al., (2019)
7. I. punctatus Green et al., (2020)
13. REVIEW OF LITERATURE
Sl.
No.
Species Stocking
density
Results Reference
1. L. vennamei
1500, 3000,
6000 and
9000/m3
It was found that the mean final weight was higher
in clear water culture (≈ 0.64, 0.41, 0.31, 0.17
g/org at SD of 1500, 3000, 6000, 9000 org/m3
respectively) than in BFT system. Survival range of
85.0- 98.4% was seen at the four stocking
densities.
Leal et al.,
(2015)
2.
Tilapia 166, 333 and
600 org/m3
Tilapia cultured at stocking densities 166 org/m3
and 333 org/m3 showed higher final body weight
and survival. The digestive enzymes, immune
response and antioxidant abilities were all
observed to be depressed for fish reared at 600
org/m3 and control group.
Liu et al.,
(2018)
14. REVIEW OF LITERATURE
Sl.
No.
Species Stocking
density
Results Reference
3.
Common
carp
6 kg/m3 CW, 12
kg/m3 CW,
6kg/m3 BFT, 12
kg/m3 BFT
The final fish density was highest in BFT 12
(25.41 ± 0.48 kg/m3). The total NH3-N
concentration was found significantly lower in
BFT groups.
Adineh et al.,
(2019)
4.
Rhamdia
quelen
10, 20 and 30
g/L
They found D30 group fishes showed
improved growth performance and lower fat
deposition in the fillet. Aspartate
aminotransferase levels and superoxide
dismutase activity were less in liver of R.
quelen reared at higher SD.
Battisti et al.,
(2020)
15. REVIEW OF LITERATURE
Sl.
No.
Species Stocking
density
Results Reference
5.
IMC 4.28, 8.57
and 12.85
fish/m3
IMC under biofloc system stocked at
4.28 fish/m3 showed higher water
quality and floc formation and the
average concentration of NH4-N, NO2-
N, NO3-N of 0.61 mg/L, 0.35mg/L and
1.8 mg/L respectively was recorded
lowest than other stocking densities.
Deb et al.,
(2020)
6.
P.
semisulcatus
10,20,30,40
shrimps
0.24m-2
They found that P. semisulcatus showed
better weight gain and survival with a
feeding rate of 6% Bw/d and 20
shrimps 0.24m-2 SD in BFT.
Kaya et al.,
(2020)
16. REVIEW OF LITERATURE
Sl.
No.
Species Stocking
density
Results Reference
7.
Nile Tilapia 20, 40, 60 and
80 no./m3
Based on daily weight gain and
survival, stocking densities of 20
and 40 no./m3 was found better
than others.
Manduca et al.,
(2020)
8.
Nile Tilapia 18.75, 37.50,
56.25, 75.00
fish/m3
The highest daily weight gain
(2.84 g/d) was obtained from low
stocking density. No significant
differences were found in body
composition. They concluded
that around 33 fish/m3 showed
higher profitability than others.
Manduca et al.,
(2020)
18. EXPERIMENTAL ANIMAL ANDFEEDING
Source:
Department of
Aquaculture/ local
farms
Advanced fry (20
days old)
Acclimatized in
clear water
Round the clock
aeration
Feeding
commercial diet
with 36% protein
19. EXPERIMENTAL DESIGN
CW3 BFT3
BFT6
CW9 BFT9
CW6 Clearwater
controls
Biofloc
treatments
Three different stocking levels of 3, 6 & 9 fishes/L will be evaluated in
triplicates
20. Levels of stocking densities designed were based on our previous experiment
(Debbarma et al., (2020), where a level of 6.4 no/L was used.
• Locally available molasses (50% carbon) will be used as an additional source
of carbon because it mainly consists of sugars.
C/N ratio of 15-20:1 (to be ascertained initially depending on the preference of
biofloc and consumption rate by experimental fish) will be created based on TAN.
• Raw salt will be used to improve flocculation in the system.
21. The experiment will be carried out for 70 days.
• Feeding will be done 5 times daily and level of feeding will be adjusted as the
culture progresses, depending on floc volume generated.
In biofloc based treatments, 5% water exchange will be done daily
whereas for clear water based treatments, it will be 40% once in 3
days.
• Weekly record of sampling data will be maintained.
22. Waterquality parameters
Sl. No. Parameters Methods Frequency
1. Water temperature Pro DO multi-parameter kit Daily
2. pH Digital pH meter Daily
3. Total Dissolved Solids TDS meter, Hanna Instruments Daily
4. Dissolved oxygen Pro DO multi-parameter kit Daily
5. Alkalinity APHA, 1995 Weekly
6. Total ammonia APHA, 1995 Weekly
7. Nitrite N APHA, 1995 Weekly
8. Nitrate N APHA, 1995 Weekly
9. Floc volume Imhoff cone Weekly
10. Chlorophyll-a APHA, 1995 Weekly
23. Zoo- technical Indices
Sl. No. Indices Formula
1. Weight gain Final average gain - Initial average weight
2. Feed Conversion Ratio
(FCR)
Total feed used (dry weight) / total weight of the
harvested fish (wet weight)
3. Protein Efficiency Ratio
(PER)
Net weight gain (g)/Protein applied in the feed (g)
4. Survival rate (Number of fish survived at the end of 70 days) /
Number stocked) × 100
24. Hematological and Biochemical Indices
Sl. No. Indices Method
1. Red Blood Cells (RBC) Ref?
2. White Blood Cells Ref?
3. Hemoglobin (Hb) Ref?
4. Hematocrit (Ht) Ref?
5. Mean Corpuscular Volume (MCV) Ref?
11. Lysozyme Method of Anderson and Siwiki
(1995)
12. Respiratory Burst Activity ?????
25. Stress biomarkers and Digestive enzymes
Sl.
No
.
Indices Method
1. Cortisol
2. Glutathione-S-
transferase (GST)
3. Catalase (CAT) Method of
Takahara et al.,
(1960)
4. Superoxide
dismutase (SOD)
Method of Misra
and Fridovich
(1972)
Sl.
No.
Indices Method
1. Protease activity Casein digestion
method
2. Amylase activity Dinitro-salicyclic-acid
(DNS) method
3. Lipase activity Cherry and Crandall
Method
4. Trypsin activity
26. Proximate analysis
Sl. No. Indices Method
1. Moisture AOAC, 2005
2. Ash AOAC, 2005
3. Crude protein AOAC, 2005
4. Crude lipid AOAC, 2005
27. data ANALYSIS
The observed data will be statistically analyzed using SPSS
software (Version 25.0) and data presented as Mean ± S.D.
28. TIME SCHEDULE
Activity Nov Dec Jan Feb March April May June July Aug Sep Oct
Experiment
al set-up
*
Test and
Observatio
n
* * * *
Result and
discussion
*
Analysis ,
report
preparation
and
submission
* * * * * *
29. EXPECTED OUTCOME
The findings from the proposed experiment will be helpful to
optimize the level of stocking for O. bimaculatus with a emphasis
on growth and physiological status in a biofloc system.
30. REFERENCES
• Adineh, H., Naderi, M., Hamidi, M.K., Harsij, M.(2019). Biofloc technology improves growth, innate immune
responses, oxidative status and resistance to acute stress in common carp under high stocking density. Fish and
shellfish immunology. 440-448.
• Azim, M.E., Little, D.C. (2008). The biofloc technology (BFT) in indoor tanks: water quality, biofloc composition,
and growth and welfare of Nile tilapia (Oreochromis niloticus). Aquaculture. 283, 29–35.
• Battisti, E.K., Rabaioli, A., Uczay, J., Sutili, E.J., Lazzari, R., (2020). Effect of stocking density on growth
,haematological and biochemical parameters and antioxidant status of silver catfish cultured in biofloc system.
Aquaculture.
• Crab, R. (2010). Bioflocs technology: an integrated system for the removal of nutrients
and simultaneous production of feed in aquaculture. PhD thesis, Ghent University. 178 pp.
• Deb, S., Noori, M.T., Rao, P.S.(2020). Application of biofloc technology for IMC culture along with water quality
management. Aquacultural Engineering.
• De Schryver, P., Crab, R., Defoirdt, T., Boon, N., Verstraete, W. (2008). The basics of bioflocs technology: The
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31. • De Schryver, P., Sinha, A.K., Baruah, K., Verstraete, W., Boon, N., De Boeck, G., Bossier, P. (2010). Poly-beta-
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