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International Journal of Recent Scientific Research Vol. 4, Issue, 7, pp.1050 1055, July, 2013 ISSN: 0976-3031
RESEARCH ARTICLE IMPACT OF PROBIOTICS ON CULTURE PONDS OF PENAEID SHRIMPS, AT EAST COAST OF ANDHRA PRADESH, INDIA
* 1Srinivas,
1Department
of Zoology, College of Science and Technology, Andhra University, Visakhapatnam-530 003 2Department of Marine Living Resources, Andhra University ABSTRACT
The current study was aimed to focus on the probiotics effect on shrimp culture ponds. The study evaluates the probiotics effects on three ponds one control pond and two experimental ponds during two consecutive years 2009 to 2010. The TVC count in water and sediments in Pond A and Pond B in summer and winter for the year 2009 was recorded as 2.33 10 0.16, 2.08 10 0.12, 2.68 10 0.15, 2.43 10 0.23, 2.33 10 0.30, 1.75 10 0.16, 2.77 10 0.31, 2.02 10 0.15.and the TVC count in water and sediments in Pond A and Pond B in summer and winter for the year 2010 was recorded as 2.85 10 0.19, 0.35 10 0.53, 3.65 10 0.21, 1.32 10 0.12 ,0.29 10 0.47, 0.27 10 0.44, 2.67 10 0.35, 0.19 10 0.15 at 120 days of culture period. The maximum TVC count was noticed in culture pond was 94 10 0.30, 26.5 10 0.19 during summer 2009 and 2010 respectively. The statistical analysis of the present study revealed that there was a significant difference in Vibrio count was observed in between control and experimental ponds. Copy Right, IJRSR, 2013, Academic Journals. All rights reserved.
ARTICLE INFO
Article History:
Received 16th, June, 2013 Received in revised form 28th, June, 2013 Accepted 17th, July, 2013 Published online 30th July, 2013
INTRODUCTION
In aquaculture environment the bottom gets deteriorated by left over feed due to excess feeding, fecal matter and dead algae. Anaerobic conditions set in due to oxygen depletion and in turn causes increase in the levels of carbon dioxide and decrease in pH levels. The management of the culture pond without causing stress to shrimp is the right approach for successful crop. The major functions of the probiotics are to degrade organic waste in pond bottom, reduced accumulated bottom sludge, and accelerate the removal of hydrogen sulphide, ammonia, nitrites in pond water and suspended solids reduction. Microbial diseases occur at all stages of shrimp aquaculture and are responsible for considerable economic losses (Leightner, 1996; Moriarty, 1998; Sung et al., 2001). Probiotics are scientifically blended concentration of selected, adopted and cultured bacterial formulations plus enzymes and special buffers fermented which cereal and mineral substrate for use in shrimp farming (Ravichandran and Shaick Jalaluddin, 1999). The use of probiotics in aquaculture may produce various beneficial effects has been proven beyond doubt (Balcazar et al., 2006; Kesarcodi-Watson et al., 2008). The research on use of probiotics in aquatic animals is increasing with the demand for environment friendly aquaculture (Gatesoupe, 1999). Improved water quality has especially been associated with probiotics it has been reported that use of Bacillus sp. improved water quality, survival and growth rates and the health status of the juvenile Penaeus monodon and reduced the pathogenic Vibriosis (Dalmin et al.,
2001). Use of commercial probiotics in Penaeus vannamei ponds can reduce concentrations of nitrogen and phosphorus and increase the shrimp yields (Wang et al., 2005). Recently, Nimrat et al., (2011) developed an effectively novel probiotic for rearing of L. vannamei through microencapsulated or freeze-dried forms of microbial probiotics: bacteria, yeast and microalgae. The current study has taken up by application of probiotics in the shrimp culture pond ecosystem in consideration of frequency and dosage of probiotics on pond water and sediments in one control and two experimental ponds both in summer and winter seasons during two consecutive years 2009 to 2010.
* Corresponding author: Srinivas, R Department of Zoology, College of Science and Technology, Andhra University, Visakhapatnam-530 003
International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 1050 - 1055, July, 2013 TCBS (Thiosulphate Citrate Bilesucrose Agar) medium under strict and hygienic microbiological laboratory conditions at M/s. Lotus Ganges Laboratories, Visakhapatnam. The water parameters of salinity, pH, temperature, observed and maintained during the study period of two years in both summer and winter crops of both the seasons from the culture ponds. The water parameters were observed at the study ponds by electronic meters. The salinity readings were taken by ATAGO refractometer, the temperature was recorded by the Thermometer, and the pH readings were taken by Hanna pH Pen - Scan-2. Statistical analysis of the data The statistical package used for interpreting the available data is microcal-origin pro 8.0 Version for evaluation of the total Vibrio count (TVC) by taking the values from the average data drawn during study period. The results were evaluated statistically to observe the decrease of total Vibrio count with application of probiotics in the experimental ponds and without probiotics in the control ponds. Significant reduction of the TVC in the pond water and sediment was observed. of culture. These experimental ponds A & B were harvested normally at 28.0 g & 29.0 g on 125th day respectively. (Table 1, Fig 1).
Fig 2 Total Vibrio Count of culture ponds at Amplam during winter 2009
RESULTS
The present experiment was carried out on the frequency and dose of application of the soil & water probiotics and the effect on the pond water & sediment by recording the results of one control and two experimental ponds with respect to the total Vibrio count (TVC) observed periodically before stocking of post larvae and at every 30days interval in the crop un till the harvest. In the present study at Amplam during the culture period in the summer crop of year 2009, the physicochemical parameters such as salinity, pH, and temperature of the study ponds ranged between 16-22 %, 7.9-8.5, and 3133C respectively. Total Vibrio count (TVC) of the pond water noticed as 2.3310 0.16 cfu ml-1 and 2.0810 0.12 cfu ml-1 in the experimental ponds A & B respectively at 120 days of culture. In the control pond, the TVC is recorded as 1.5710 0.40 cfu ml-1 and 2.4010 0.13 cfu mg-1 in the pond water and sediment respectively at 90 days of culture (DOC).
Fig 3 Total Vibrio Count of culture ponds at Amplam during summer 2010
The physico-chemical parameters in the winter crop during the year 2009 such as salinity, pH, and temperature of the study ponds ranged between 15-20 , 7.5-8.6, and 25-28C respectively. The TVC results recorded as 3.3710 0.80 cfu ml-1, 2.3310 0.30 cfu ml-1 and 1.7510 0.16 cfu ml-1 in water of control, experimental pond A & pond B at 90 days & 120 days of culture respectively .
Fig 1 Total Vibrio Count of culture ponds at Amplam during summer 2009
Fig 4 Total Vibrio Count of culture ponds at Amplam during winter 2010
The shrimp P.monodon in this control pond was harvested at 22.5 g on 107th day with the incidence of white spot disease. At the same time the TVC in the in the sediment of the experimental ponds A & B, were recorded as 2.6810 0.15 cfu mg-1 and 2.4310 0.23 cfu mg-1 respectively at 120 days
Where as in the sediment the total Vibrio count is noticed as 3.6810 0.20 cfu mg-1, 2.7710 0.31 cfu mg-1 and 2.0210 0.15 cfu mg-1 in control, experimental pond A & pond B respectively at 90 & 120 days of culture.
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International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 1050 - 1055, July, 2013 Table 1 Total Vibrio Count of culture ponds at Amplam during summer 2009 Control pond
S.No 1 2 3 4 5 Days of culture Before stocking 30 60 90 120 Salinity ppt 19 20 20 19 pH 7.9 8.3 8.3 8.5 Temp C 31 32 32 32 TVC Water cfu/ml NIL 94 10 0.30 0.23 10 0.32 1.57 10 0.40 TVC Sediment cfu/mg NIL 33.7 10 0.16 0.20 10 0.19 2.40 10 0.13 -
Control pond harvested due to white spot disease at 22.5 g on 107th day
Pond A
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 19 21 20 19 16 pH 7.9 8.2 8.3 8.5 8.4 Temp C 31 32 32 32 31 TVC Water cfu/ml NIL 0.15 10 0.26 0.23 10 0.32 1.57 10 0.40 2.33 10 0.16 TVC Sediment cfu/mg NIL 0.12 10 0.19 0.20 10 0.19 2.40 10 0.13 2.68 10 0.15
Pond B
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 19 21 20 20 17 pH 7.9 8.2 8.3 8.4 8.5 Temp C 31 32 32 31 32 TVC Water cfu/ml NIL NIL 0.18 10 0.13 0.34 10 0.14 2.08 10 0.12 TVC Sediment cfu/mg NIL 0.10 10 0.35 0.15 10 0.24 2.38 10 0.19 2.43 10 0.23
Table 2 Total Vibrio Count of culture ponds at Amplam during winter 2009 Control Pond
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 16 18 19 19 pH 7.5 7.9 8.2 8.6 Temp C 27 26 25 25 TVC Water cfu/ml NIL 0.20 10 0.48 2.35 10 0.30 3.37 10 0.80 TVC Sediment cfu/mg NIL 2.82 10 0.75 2.95 10 0.39 3.68 10 0.20 -
Pond A
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 15 17 18 19 20 pH 7.9 8.2 8.1 8.4 8.3 Temp C 28 27 27 25 25 TVC Water cfu/ml NIL 0.11 10 0.10 0.31 10 0.52 1.78 10 0.38 2.33 10 0.30 TVC Sediment cfu/mg NIL NIL 0.27 10 0.29 2.45 10 0.27 2.77 10 0.31
Pond B
S.N0 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity Ppt 15 17 18 19 20 pH 8.1 7.9 8.2 7.9 8.1 Temp C 28 27 26 26 25 TVC Water cfu/ml NIL 0.10 10 0.13 0.22 10 0.15 1.03 10 0.45 1.75 10 0.16 TVC Sediment cfu/mg NIL NIL 0.19 10 0.22 1.00 10 0.15 2.02 10 0.15
Table 3 Total Vibrio Count of culture ponds at Amplam during summer 2010 Control Pond
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity Ppt 19 23 25 29 31 pH 8.2 8.5 8.7 8.3 8.5 Temp C 28 31 32 32 28 TVC Water cfu/ml NIL 0.28 10 0.41 1.55 10 0.19 17.8 10 0.46 53.3 10 0.77 TVC Sediment cfu/mg NIL 0.39 10 0.43 2.55 10 0.37 2.95 10 0.16 70.8 10 1.11
Pond A
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity Ppt 19 22 26 28 30 pH 8.5 8.5 8.7 8.8 8.7 Temp C 29 30 31 31 29 TVC Water cfu/ml NIL 0.16 10 0.19 NIL 0.29 10 0.61 2.85 10 0.19 TVC Sediment cfu/mg NIL 0.27 10 0.46 NIL 0.30 10 0.36 3.65 10 0.21
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International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 1050 - 1055, July, 2013 Pond B
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 19 22 26 28 30 pH 8.5 8.4 8.6 8.8 8.7 Temp C 28 30 31 32 28 TVC Water cfu/ml NIL NIL NIL NIL 0.35 10 0.53 TVC Sediment cfu/mg NIL 0.16 10 0.19 NIL NIL 1.32 10 0.12
Table 4 Total Vibrio Count of culture ponds at Amplam during winter 2010 Control Pond
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 14 17 18 pH 7.4 8.6 8.9 Temp C 28 27 26 TVC Water cfu/ml NIL 8.97 10 0.38 26.5 10 0.19 TVC Sediment cfu/mg NIL 2.65 10 0.19 5.12 10 0.33 -
Pond A
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 15 15 16 17 17 pH 7.9 8.5 8.4 8.3 8.5 Temp C 28 27 26 26 26 TVC Water cfu/ml NIL 0.14 10 0.22 NIL 0.21 10 0.25 0.29 10 0.47 TVC Sediment cfu/mg NIL NIL NIL 2.17 10 0.50 2.67 10 0.35
Pond B
S.No 1 2 3 4 5 DOC BEFORE 30 DOC 60 DOC 90 DOC 120 DOC Salinity ppt 15 16 16 17 16 pH 8.4 8.2 8.5 8.4 8.6 Temp C 28 27 26 26 25 TVC Water cfu/ml NIL NIL NIL 0.12 10 0.14 0.27 10 0.44 TVC Sediment cfu/mg NIL NIL NIL 0.17 10 0.14 0.19 10 0.15
The shrimp P.monodon cultured in the control pond is harvested at 16.0 g on 91st day with white spot disease and the experimental ponds A & B were harvested normally at 26.0 g & 27.0 g on 124th day & 125th day respectively. (Table 2, Fig 2). The salinity, pH, and temperature of the study ponds were recorded between 19-31 , 8.2-8.8, and 28-32C respectively in the summer crop of year 2010. The total Vibrio colonies in the pond water were noticed as 53.310 0.77 cfu ml-1, 2.8510 0.19 cfu ml-1 and 0.3510 0.53 cfu ml-1 in control, experimental pond A & pond B respectively. In the sediment of the ponds the TVC is recorded as 70.810 1.11 cfu mg-1, 3.6510 0.21 cfu mg-1 and 1.3210 0.12 cfu mg-1 in control, experimental pond A & pond B respectively. All the above TVC readings were recorded at 120 days of culture period. Here in this crop the shrimp P.monodon in the control pond, experimental pond A & pond B were harvested normally at 31.5 g, 35.0 g, and 36.5 g on 137th day, 139th day & 141st day respectively. (Table 3, Fig 3). In the winter crop of the year 2010, The parameters like salinity, pH, and temperature of the study ponds ranged between 14-18 , 7.48.9, and 25-28C respectively. The total Vibrio count in the pond water was recorded as 26.510 0.19 cfu ml-1, 0.2910 0.47 cfu ml-1 and 0.2710 0.44 cfu ml-1 in control, experimental pond A & pond B respectively in the sediment the readings were noticed as 5.1210 0.33 cfu mg-1, 2.6710 0.35 cfu mg-1
and 0.1910 0.15 cfu mg-1 respectively in control, experimental pond A & pond B. The TVC values were observed at 60 days in control pond and at 120 days in experimental pond A & pond B. The control pond is harvested on 64th day at 7.0 g which is harvested due to the cross contamination by white spot disease carried from the neighboring farm. In this crop of shrimp P.monodon the experimental pond A & pond B were harvested normally at 30.5 g & 32.0 g on 129th day & 130th day respectively. (Table 4, Fig 4).
DISCUSSION
In present study great focus was done on the frequency and dosage of application of the soil & water probiotics and the effect on the pond water and sediment. The results of the present study have shown that positive benefits of probiotics to water and bottom soil quality. Boyd et al., (1994) found that the accumulated sediments, in shrimp ponds in Thailand were largely composed of mineral soil eroded from the pond banks. Smith 1996 found that 70-80% of the accumulated sediments in prawn ponds were a mixture of quartz, kaolinite and micaillite mineral 5-10%, amorphous iron and aluminum & silicon oxides, 5-10% organic matter and volatile compounds and elevated levels of trace elements. Smith (1996) did not find a relationship between sediment accumulation and factors such as organic inputs, pond management or productivity. Hopkins et al., (1994) working in plastic lined ponds, suggested that the source of the accumulated sediments in a pond are uneaten food faces, decaying plankton, air borne debris, eroded soil
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International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 1050 - 1055, July, 2013 and microorganisms. Smith (1993) found that the accumulated sediments in prawn ponds contained large amounts of silica and that diatoms were significant part of the sediments. Accumulated sediments in shrimp ponds are highly reduced (Guo, 1986) enriched in organic matter (Boyd and Tucker 1992; Martin et al., 1998). Ram et al., (1982) found that the density of aerobic and anaerobic bacteria in the pond bottom soil is 2 to 4 orders of magnitude higher than the density of these groups in the water column. Moriarty (1996) found that the bacterial count in the sediments of shrimp ponds fertilized with chicken manure was 4x1013 bacteria m-2 which was 2 to 3 orders of magnitude higher than in the water column. Burford et al., (1998) reported a bacteria count of 15.5x109 cellsg-1 in the centre of shrimp pond where sludge accumulated, and 8.1x109 cellsg-1 at the periphery of the pond. The transience of aquatic microbes may legitimate the extension of the probiotic concept to living microbial preparations use to treat aquaculture ponds. Moriarty (1998) proposed to extend the definition of probiotics to microbial water additives. However these extensions would make too vague definition of Tannock (1997). In 1991 Porubcan reported on two attempts at bacterial treatments to improve water quality and production yield of Penaeus monodon. For this Porubcan made floating biofilters, pre-inoculated with nitrifying bacteria decreased the amounts of ammonia and nitrite in the rearing water. This treatment increase shrimp survival. The introduction of Bacillus spp., in proximity to pond aerators reduced chemical oxygen demand (COD) and increased shrimp harvest (Porubcan, 1991). In the present study by application of probiotics the two experimental ponds showed positive results in the shrimp production when compared to the control ponds. The water samples of all the ponds were analyzed periodically for total Vibrio colonies and the findings were revealed and the total Vibrio colonies were increased in control ponds of both seasons in the two years of study period. There is significant reduction in the total Vibrio colonies in the experimental ponds in both pond water and pond sediment. It is evident from the obtained results from the present study that the reduction of the total Vibrio colonies with increased dosage of the pond probiotic and also by increase the frequency of application from 15 days duration to 7 days duration. The positive bacteria in the probiotics reduced the proliferation of the pathogenic bacteria in all the ponds where the application of probiotics is followed. At the same time due to the probiotic application the toxic gases of the pond bottom were reduced and the oxidation of the organic matter is observed in the culture ponds.
References
Balczar, J.L., de Blas, I., Ruiz-Zarzuela, I., Cunningham, D., Vendrell, D., Mzquiz, J.L. 2006. The role of probiotics in aquaculture. Vet. Microbiol., 114: 173186. Boyd, C.E. and Tucker C.S. 1992. Water quality and pond soil analysis for aquaculture. Auburn University, Alabama. 183. pp. Boyd, C.E., Tanner, M., Madkour, E., Masuda, K. 1994: Chemical charecterstics of bottom soils, from fresh water and brakish water aquaculture ponds. J.world aquatic Soc., 25: 517-534. Burford, M.A., Peterson, E.L., Baino, J.C.F., Peterson, N.P. 1998. Bacteria in shrimp pond sediments, their role in mineralizing nutrients and some suggested sampling stratagies. Aquac. Res., 29: 843-849. Dalmin, G., Kathiresan, K., Purushothaman, A. 2001. Effect of probiotics on bacterial population and health status of shrimp in culture pond ecosystem. Indian J. Exp. Biol., 39: 939942. Gatesoupe, F.J., 1999. The use of probiotics in aquaculture. Aquaculture., 180: 147165. Guo, S. 1986. Determination of oxidation reduction potential in penaied shrimp prawn growing up ponds. Mpr.Fish Res., 7: 89-93. Hopkins, J.S., Sandifer, P.A., Browdy, C.L. 1994. Sludge management in intensive pond culture of shrimp: Effect of management regime on water quality, sludge characteristics, nitrogen extinction, and shrimp production. Aqua-cultural Engineering., 13:11-30. Kesarcodi-Watson, A., Kaspar, H., Lategan, M.J., Gibson, L. 2008. Probiotics in aquaculture: The need, principles and mechanisms of action and screening processes. Aquaculture., 274: 114. Lightner, D.V. 1996. A Handbook of Shrimp Pathology and Diagnostic Procedures for Diseases of Cultured Penaeid Shrimp. World Aquaculture Society, Baton Rough, Lour Siena, USA, pp. 1 72. Martin, J.L.M., Vern Y., Guelorget, O., Pham, D. 1998. Shrimp rearing stocking density growth, impact on sediment, water put and their relationships studied through nitrogen budget in rearing ponds. Aquaculture., 164: 135- 149. Moriarty, D.J.W., 1996. Microbial biotechnology: a key ingredient for sustainable aquaculture. INFOFISH International., 4(96):29-33. Moriarty, D.J.W., 1998. Control of luminous Vibrio species in penaeid aquaculture ponds. Aquaculture., 164: 351 358. Nimrat, S., Boonthai, T., Vuthiphandchai, V. 2011. Effects of probiotic forms, compositions of and mode of probiotic administration on rearing of Pacic white shrimp (Litopenaeus vannamei) larvae and postlarvae. Anim. Feed Sci. Technol., 169: 244258. Porubcan, R.S. 1991. Reduction in Chemical oxygen demand and improvement in Penaeus monodon yield in ponds inoculated with aerobic Bacillus bacteria. Program and Abstracts of the 22nd Annual Conference and Exposition, 16-20 June 1991, San Juan, Puerto Rico. World Aquaculture Society. Ram, N., Zur, O., Avenimalech, Y., 1982. Microbial changes occurring at the sediment interface in an intensively stocked and fed fish pond. Aquaculture., 27: 63-72. Ravichandran, R., Shaick, J.R. and Jalaluddin, R. 1999. Bacterial amendment-technology for disease free shrimp culture. Paper presented at the National seminar on development & transfer of fisheries technology, Tuticorin (TN), India.
CONCLUSION
The results of the current study suggested that the decreased TVC count was recorded when the probiotics were used for the current experimentation for two consecutive years of the study both in summer and winter seasons. The significant difference in TVC values were noticed between control and experimental ponds.
Acknowledgements
The author express thank to the Head, Department of Zoology, Andhra University for carried out this research work
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International Journal of Recent Scientific Research, Vol. 4, Issue, 7, pp. 1050 - 1055, July, 2013 Smith, P.T. 1993. Prawn farming in Australia sediment is a major issue. Aust. Fish., 52: 29-32. Smith, P.T. 1996. Physical and chemical characteristics of sediments from prawn farms and mangrove habitats on the Clarence River. Australia Aquaculture., 146: 47-83. Sung, H.H., Hsu, S.F., Chen, C.K., Ting, Y.Y., Chao, W.L. 2001. Relationship between disease outbreaks in cultured tiger shrimp (Penaeus monodon) and the composition of Vibrio communities in pond water and shrimp hepatopancreas during cultivation. Aquaculture., 192: 101 110. Tannock, G.W. 1997. Modification of the normal micro biota by diet, stress, antimicrobial agents, and probiotics. In: Mackie, R. I., With, B.A., Isaacson, R.E. (Eds.), Gastrointestinal Microbiology, Vol. 2, Gastrointestinal and Host Interactions. Chapman and Hall Microbiology Series, International Thomson Publishing, New York, pp. 434 465. Wang, Y.B., Xu, Z.R., Xia, M.S. 2005. The effectiveness of commercial probiotics in Northern White Shrimp (Penaeus vannamei L.) ponds. Fish. Sci., 71: 10341039.
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