Fish Physiology and Biochemistry

, Volume 35, Issue 3, pp 467–478 | Cite as

Effect of a probiotic bacterium Bacillus circulans PB7 in the formulated diets: on growth, nutritional quality and immunity of Catla catla (Ham.)

  • Partha Bandyopadhyay
  • Pradeep K. Das Mohapatra


Bacillus circulans PB7, isolated from the intestine of Catla catla, was evaluated for use as a probiotic supplement in the feeds for the fingerlings of Catla catla. The effect of supplement on the growth performance, feed utilization efficiency, and immune response was evaluated. Catla fingerlings (ave. wt. 6.48 ± 0.43 g) were fed diets supplemented with 2 × 104 (feed C1), 2 × 105 (feed C2), and 2 × 106 (feed C3) B. circulans PB 7 cells per 100 g feed for 60 days at 5% of the body weight per day in two equal instalments in triplicate treatments. The control feed (CC) was not supplemented with the B. circulans. All the feeds were isocaloric and isonitrogenous. Fish fed with feed C2 displayed better growth, significantly (P ≤ 0.05) highest RNA/DNA ratio, a lower feed conversion ratio (FCR), and a higher protein efficiency ratio (PER) than the other experimental diets. Highest carcass protein and lipid was also observed in the fish fed C2 feed compared to the others. Significantly (P ≤ 0.05), highest protease was recorded in fish fed feed C2 (47.9 ± 0.016) and lowest in fish fed feed C3 (32.10 ± 0.009), where α-amylase activity did not differ significantly (P ≤ 0.05) beyond the lowest inclusion level. ALP, ACP, GOT, and GPT in the liver of Catla catla were the highest (P ≤ 0.05) in fish fed C2 feed. The highest TSP, albumin, and globulin was observed in fish treated with C2 feed after 60 days feeding trial, but the lowest glucose level was observed in the same treatment. After the feeding trial, the non-specific immunity levels and disease resistance of fish were also studied. Phagocytic ratio, phagocytic index, and leucocrit value were the highest in fish fed feed C2. After the feeding trial, the fish were challenged for 10 days by bath exposure to Aeromonas hydrophila (AH1) (105c.f.u. ml−1 for 1 h, and, after 7 days, 107c.f.u. ml−1 for 1 h). Highest survival percentage was observed in fish fed with feed C2 compared with only 6.66% in the controls, which indicated the effectiveness of B. circulans PB 7 in reducing disease caused by A. hydrophila.


Bacillus circulans Probiotics Catla catla 



The first author is highly thankful to Indian Council of Agricultural Research, New Delhi, India for financial assistance. The author is also thankful to Head of the Department, Department of Zoology and Hon’ble Vice-Chancellor, Vidyasagar University for providing necessary facilities.


  1. Ajitha S, Sridhar M, Sridhar N, Singh ISB, Varghese V (2004) Probiotic effects of lactic acid bacteria against Vibrio alginolyticuys in Penaeus (Fenneropenaeus) indicus (H. Milne Edwards). Asian Fish Sci 17:71–80Google Scholar
  2. Akinbowale OL, Peng H, Barton MD (2006) Antimicrobial resistance in bacteria isolated from aquaculture sources in Australia. J Appl Microbiol 100:1103–1113. doi: 10.1111/j.1365-2672.2006.02812.x PubMedCrossRefGoogle Scholar
  3. Alcaide E, Blasco MD, Esteve C (2005) Occurrence of drug-resistant bacteria in two European eel farms. Appl Environ Microbiol 71:3348–3350. doi: 10.1128/AEM.71.6.3348-3350.2005 PubMedCrossRefGoogle Scholar
  4. Alfaro J, Lawrence AL, Lewis D (1993) Interaction of bacteria and male reproductive-system blackening disease of captive Penaeus setiferus. Aquaculture 117:1–8. doi: 10.1016/0044-8486(93)90117-H CrossRefGoogle Scholar
  5. AOAC (1990) Official methods of analysis, 15th edn. S Williams (ed) AOAC, Virginia, 1298 ppGoogle Scholar
  6. APHA-AWWA-WPCF (1998) Standard methods for the examination of water and wastewater, 20th edn. Washington DCGoogle Scholar
  7. Austin B, Austin DA (1999) Bacterial fish pathogens: disease of farmed and Wild Fish, 3rd edn. Springer-Praxix, GodalmingGoogle Scholar
  8. Austin B, Stuckey LF, Robertson PAW, Effendi I, Griffith DRW (1995) A probiotic strain of Vibrio algilolyticus effective in reducing diseases caused by Aeromonas salmonicida, Vibrio anguillarum and Vibrio ordalii. J Fish Dis 18:93–96. doi: 10.1111/j.1365-2761.1995.tb01271.x CrossRefGoogle Scholar
  9. Bairagi A, Sarkar GK, Sen SK, Ray AK (2004) Evaluation of the nutritive value of Leucaena leucocephala leaf meal, inoculated with fish intestinal bacteria Bacillus subtilis and Bacillus circulans in formulated diets for rohu, Labeo rohita (Hamilton) fingerlings. Aquat Res 35:436–446. doi: 10.1111/j.1365-2109.2004.01028.x CrossRefGoogle Scholar
  10. Balebona MC, Zorrilla I, Morinigo MNA, Berrego JJ (1998) Survey of bacterial pathologies effect in farmed gilthead sea bream (Sparus aurata L.) in South Western Spain from 1990–1996. Aquaculture 166:19–35. doi: 10.1016/S0044-8486(98)00282-8 CrossRefGoogle Scholar
  11. Bandyopadhyay P, Swain SK, Mishra S (2005) Growth and dietary utilisation in goldfish (Carassius auratus Linn) fed diets formulated with various local agro-produces. Bioresour Technol 96:731–740. doi: 10.1016/j.biortech.2004.06.018 PubMedCrossRefGoogle Scholar
  12. Bazaz MM, Keshavanath P (1993) Effect of feeding levels of sardine oil on growth, muscle composition and digestive enzyme activities of mahseer Tor khudree. Aquaculture 115:111–119. doi: 10.1016/0044-8486(93)90362-3 CrossRefGoogle Scholar
  13. Bernfeld P (1955) Amylase-oc and -ß. In: Colowick SP, Kaplan NO (eds) Methods of enzymology, vol 1. Academic Press, New York, pp 141–158CrossRefGoogle Scholar
  14. Blaxhall PC, Daisley KW (1973) Routine haematological methods for use in fish blood. J Fish Biol 5:771–781. doi: 10.1111/j.1095-8649.1973.tb04510.x CrossRefGoogle Scholar
  15. Bramley TA (1974) Treatment of immature mice with gonadotropins. Effects on some enzymatic activities of unfractioned ovarian homogenates. J Biochem 140:451–460Google Scholar
  16. Brown JH (1989) Antibiotics: their use and abuse in aquaculture. World Aquaculture 20:34–42Google Scholar
  17. Buckley LJ (1980) Changes in ribonucleic acid, deoxyribonucleic acid and protein content during ontogenesis in winter flounder Pseudopleuronectes amouricauses and effect of starvation. US Fish Bull 77:703–708Google Scholar
  18. Chang CI, Liu WY (2002) An evaluation of two probiotic bacterial strains, Enterococcus faecium SF 68 and Bacillus toyoi, for reducing edwardsiellosis in cultured European eel, Anguilla anguilla L. J Fish Dis 25:311–315. doi: 10.1046/j.1365-2761.2002.00365.x CrossRefGoogle Scholar
  19. Das BK, Samal SK, Samantaray BR, Sethi S, Pattnaik P, Misha BK (2006) Antagonistic activity of cellular components of Pseudomonas species against Aeromonas hydrophila. Aquaculture 253:17–24. doi: 10.1016/j.aquaculture.2005.01.028 CrossRefGoogle Scholar
  20. Douillet PA, Langdon CJ (1994) Use of probiotic for the culture of larvae of the Pacific oyster (Crassostrea gigas Thunberg). Aquaculture 119:25–40. doi: 10.1016/0044-8486(94)90441-3 CrossRefGoogle Scholar
  21. Duncan DB (1955) Multiple range and multiple F-tests. Biometrics 11:1–42. doi: 10.2307/3001478 CrossRefGoogle Scholar
  22. Fuller R (1987) A review, probiotics in man and animals. J Appl Bacteriol 66:365–378Google Scholar
  23. Fuller R (1992) History of development of probiotics. In: Fuller R (ed) Probiotics: the scientific basis. Chapman and Hall, New York, pp 1–8Google Scholar
  24. Garriques D, Arevalo G (1995) An evaluation of the production and the use of a live bacterial isolate to manipulate the microbial flora in the commercial production of Penaeus vannamei post larvae in Ecuador. In: Browdy CL, Hopkins JS (eds) Swimming through troubled water. Proceedings of the special session on shrimp farming, Aquaculture, 1995. World Aquaculture Society, Baton Rouge, pp 53–59Google Scholar
  25. Gatesoupe FJ (1991) The effect of three strains of lactic bacteria on the production rate of rotifers, Brachionus plicatilis, and their dietary value for larval turbot, Scophthalmus maximus. Aquaculture 96:335–342. doi: 10.1016/0044-8486(91)90162-Z CrossRefGoogle Scholar
  26. Gatesoupe FJ (1994) Lactic acid bacteria increased the resistance of turbot larvae, Scophthalmus maximus, against pathogenic vibrio. Aquat Living Resour 7:277–282. doi: 10.1051/alr:1994030 CrossRefGoogle Scholar
  27. Ghosh K, Sen SK, Ray AK (2003) Supplementation of an isolated fish gut bacterium, Bacillus circulens, in formulated diets for rohu, Labeo rohita fingerlings. Bamidgeh 55:13–21Google Scholar
  28. Hirata H, Murata O, Yamada S, Ishitani H, Wachi M (1998) Probiotic culture of the rotifer Brachionus plicatilis. Hydrobiologia 387/388:495–498. doi: 10.1023/A:1017004124600 CrossRefGoogle Scholar
  29. Karunasagar I, Pai R, Malathi GR, Karunasagar I (1994) Mass mortality of Penaeus monodon larvae due to antibiotic resistant Vibrio harveyi infection. Aquaculture 128:203–209. doi: 10.1016/0044-8486(94)90309-3 CrossRefGoogle Scholar
  30. Kennedy SB, Tucker JW, Neidig CL, Vermeer GK, Cooper VR, Jarrell JL, Sennett DG (1998) Bacterial management Strategies for stock enhancement of warm water marine fish: a case study with common snook (Centropomus undecimalis). Bull Mar Sci 62:573–588Google Scholar
  31. Keshavanath P, Renuka P (1998) Effect of dietary L-carnitine supplements on growth and body composition of fingerling rohu. Labeo rohita (Ham.). Aquat Nutr 4:83–87. doi: 10.1046/j.1365-2095.1998.00052.x CrossRefGoogle Scholar
  32. Khan MA, Jafri AK (1991) Protein and nucleic acid concentration in the muscle of catfish Clarias batrachus. Asian Fish Sci 4:75–84Google Scholar
  33. Lee KK, Yu SR, Chen FR, Yang TI, Liu PC (1996) Virulence of Vibrio alginolycicus isolated from diseased tiger prawn Penaeus monodon. Curr Microbiol 32:229–231. doi: 10.1007/s002849900041 PubMedCrossRefGoogle Scholar
  34. Mohanty SN, Swain SK, Tripathi SD (1996) Rearing of catla (Catla catla Ham.) spawn on formulated diets. J Aquat Trop 11:253–258Google Scholar
  35. Moriarty DJW (1998) Control of luminous Vibrio species in penaeid aquaculture ponds. Aquaculture 164:351–358. doi: 10.1016/S0044-8486(98)00199-9 CrossRefGoogle Scholar
  36. Munro HN, Fleck A (1969) Analysis of tissue and body fluids for nitrogen constituents in mammalian protein metabolism. In: Munro HN (ed) Mammalian protein metabolism, vol 3. Academic Press, New York, pp 433–525Google Scholar
  37. Olsson JC, Joborn K, Westerdhal A, Blomberg L, Kjelleberg S, Conway PL (1998) Survival persistence and proliferation of Vibrio anguillarum in juvenile turbot Scophthalmus maximus (L.), in intestine and faeces. J Fish Dis 21:1–9. doi: 10.1046/j.1365-2761.1998.00327.x CrossRefGoogle Scholar
  38. Park JH, Jeong HD (1996) Enhanced resistance against Edwardsiella tarda infection in tilapia (Oreochromis niloticus) by administration of protein-bound polysaccharide. Aquaculture 143:135–143. doi: 10.1016/0044-8486(95)01224-9 CrossRefGoogle Scholar
  39. Patra S (2002) Efficacity of some aquatic weeds on the kinetics of food intake, growth promotion, associated physiological and biochemical changes in Indian major carps. PhD Thesis, Vidyasagar University, IndiaGoogle Scholar
  40. Queiroz JF, Boyd CE (1998) Effects of a bacterial inoculum in channel catfish ponds. J World Aquacult Soc 29:67–73. doi: 10.1111/j.1749-7345.1998.tb00300.x CrossRefGoogle Scholar
  41. Ringo E, Gatesoupe FJ (1998) Lactic acid bacteria in fish: a review. Aquaculture 160:177–203. doi: 10.1016/S0044-8486(97)00299-8 CrossRefGoogle Scholar
  42. Robertson PAW, O’Dowd C, Burrells C, Williams P, Austin B (2000) Use of Carnobacterium sp as a probiotic for Atlantic salmon (Salmo salar L) and rainbow trout (Oncorhynchus mykiss Walbaum). Aquaculture 185:235–243. doi: 10.1016/S0044-8486(99)00349-X CrossRefGoogle Scholar
  43. Rosauki SR (1993) Boehringer Mannheim Gmbh Analysis Protocol. Clin Chem 39:648Google Scholar
  44. Sahoo PK, Mukherjee SC (2001) Effect of dietary β-1, 3 glucan on immune responses and disease resistance of healthy and aflatoxin B1 induced immunocompromised rohu (Labeo rohita Hamilton). Fish Shellfish Immunol 11:683–695. doi: 10.1006/fsim.2001.0345 PubMedCrossRefGoogle Scholar
  45. SAS (1991) SAS Institute SAS® system for regression, 2nd edn. SAS Institute, Cary, NCGoogle Scholar
  46. Singh BN (1989) The digestibility of protein and energy from feedstuffs and pellated diets in mrigal, Cirrhinus mrigala (Ham.) and grass carp, Ctenopharyngodon idella (Val.). J Freshwat Biol 1:7–13Google Scholar
  47. Siwicki AK, Anderson DP, Rumsey GL (1994) Dietary intake of immunostimulants by rainbow trout affects non-specific immunity and protection against furunculosis. Vet Immunol Immunopathol 41:125–139. doi: 10.1016/0165-2427(94)90062-0 PubMedCrossRefGoogle Scholar
  48. Smoragiewicz W, Bielecka M, Babuchowski A, Boutard A, Dubau H (1993) Les probiotiques. Can J Microbiol 39:1089–1095CrossRefGoogle Scholar
  49. Snell FD, Snell CT (1971) Colorimetric methods of analysis, vol IV AAA. Van Nostrane Reinhold, New York, pp 7–145 Google Scholar
  50. Sundaryono A, Tsvtnenko E, Evans LH (1996) Digestibility studies on fisheries by products based diets of Penaeus monodon. Aquaculture 143:331–340. doi: 10.1016/0044-8486(96)01288-4 CrossRefGoogle Scholar
  51. Sutton DC, Garrick R (1993) Bacterial disease of cultured giant clam Tridacna gigas larvae. Dis Aquat Organ 16:47–53. doi: 10.3354/dao016047 CrossRefGoogle Scholar
  52. Swain SK, Rangacharyulu PV, Sarkar S, Das KM (1996) Effect of a probiotic supplement on growth, nutrient utilization and carcass composition in mrigal fry. J Aquat 4:29–35Google Scholar
  53. Swick RW, Barnstein PL, Strange JL (1965) The response of the isozymes of alanine aminotransferase to diet and hormones. J Biol Chem 240:3341–3345PubMedGoogle Scholar
  54. Wada H, Marino Y (1964) Comparative studies on glutamic-oxaloacetic transaminases from the mitochondrial and soluble fractions of mammalian tissues. In: Harris RS, Wool IG, Loraine JA (eds) Vitamins and hormones. Academic Press, New York, pp 411–444Google Scholar
  55. Wang Y-B (2007) Effect of probiotics on growth performance and digestive enzyme activity of the shrimp Penaeus vannamei. Aquaculture 269:259–264. doi: 10.1016/j.aquaculture.2007.05.035 CrossRefGoogle Scholar
  56. Wang X-H, Ji W-S, Xu H-S (1999) Application of probiotic in aquaculture. Aiken Murray (Internet)Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Partha Bandyopadhyay
    • 1
    • 2
  • Pradeep K. Das Mohapatra
    • 3
  1. 1.Aquaculture Research Unit, Department of ZoologyVidyasagar UniversityMidnaporeIndia
  2. 2.Biostadt India LimitedMumbaiIndia
  3. 3.Department of MicrobiologyVidyasagar UniversityMidnaporeIndia

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