Journal of Applied & Environmental Microbiology
ISSN (Print): 2373-6747 ISSN (Online): 2373-6712 Website: Editor-in-chief: Sankar Narayan Sinha
Open Access
Journal Browser
Journal of Applied & Environmental Microbiology. 2014, 2(5), 203-207
DOI: 10.12691/jaem-2-5-1
Open AccessArticle

Studies on Bacillus subtilis, as Potential Probiotics, on the Hematological and Biochemical Parameters of Rainbow trout, Oncorhynchus mykiss (Walbaum)

Maryam Kamgar1 and Masood Ghane1,

1Department of Microbiology, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran

Pub. Date: July 02, 2014

Cite this paper:
Maryam Kamgar and Masood Ghane. Studies on Bacillus subtilis, as Potential Probiotics, on the Hematological and Biochemical Parameters of Rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of Applied & Environmental Microbiology. 2014; 2(5):203-207. doi: 10.12691/jaem-2-5-1


The probiotic activity of Bacillus subtilis was evaluated by its effect on the hematological and biochemical factors of rainbow trout. The experience was carried out in 2 groups (Control and Treatment) and 3 replicates. In Control group, probiotic was not applied in diet but in Treatment group, B. subtilis was administered in feed at a concentration of 107 cells/g. In the day of thirty, 5 blood samples were caught from every replicate for biochemical and hematological experiments. Results showed that B. subtilis addition to diet had not effect on erythrocyte count, hemoglobin, hematocrit, mean cell volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) and there was no significant difference among two groups (p>0.05). But the leucocyte count, percent of lymphocyte, serum total protein, serum albumin, IgM and lysozyme of T group was significantly higher than that of C group (p<0.05), whereas percent of neutrophile and monocyte of C group was significantly higher than that of T group (p<0.05). The results suggest that B. subtilis can stimulate immune parameters in rainbow trout.

probiotic Bacillus subtilis rainbow trout Hematological and Biochemical factors

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Soltani, M., Munday, B.L., and Burke, C.M., The relative susceptible of fish to infections by Flexibacter columnaris and Flexibacter maritimus. Aquaculture, 140: 259-264. 1996.
[2]  Ellis, A.E., Tissue residues of chemotherapeutants in fish. Bull Eur Assoc Fish Pathology, 11:22-29. 1991.
[3]  Smith, P., Hiney, M.P., and Samuelson, O.B., Bacterial resistance to antimicrobial agents used in fish farming: a critical evaluation of method and meaning. Ann Rev Fish Dis, 4: 273-313. 1994.
[4]  Chantharasophon, K., Warong, T., Mapatsa, P., and Leelavatcharamas, V., High Potential Probiotic Bacillus Species from Gastro-intestinal Tract of Nile Tilapia (Oreochromis niloticus). Biotechnology, 10: 498-505. 2011.
[5]  WHO (World Health Organization). Report of a joint FAO/OIE/WHO expert consultation on antimicrobial use in aquaculture and antimicrobial resistance. Seoul, Republic of Korea. 2006.
[6]  Deeseenthum, S., Leelavatcharamas, V., and Brooks, J.D., Effect of Feeding Bacillus sp. As Probiotic Bacteria on Growth of Giant Freshwater Prawn (Macrobrachium rosenbergii de Man). Pakistan Journal of Biological Sciences, 10: 1481-1485. 2007.
[7]  Vivas, J., Riano, J., Carracedo, B., Razquin, B.E., Lopez-Fierro, P., and Naharro, G., The auxotrophic aroA mutant of A. hydrophila as a live attenuated vaccine against A. salmonicida infections in rainbow trout. Fish & Shellfish Immunology, 16: 193-206. 2004.
[8]  Bagheri, T., Hedayati, S.A., Yavari, V., Alizade, M., and Farzanfar, A., Growth, Survival and Gut Microbial Load of rainbow trout, Onchorhynchus mykiss (Walbaum) Fry Given Diet Supplemented with Probiotic during the Two Months of First Feeding. Journal of Fisheries and Aquatic Science, 8: 43-48. 2008.
[9]  Hung, A.T.Y., Su, T.M., Liao, C.W., and Lu, J.J. Effect of Probiotic Combination Fermented Soybean Meal on Growth Performance, Lipid Metabolism and Immunological Response of Growing-Finishing Pigs. Asian Journal of Animal and Veterinary Advances, 3: 431-436. 2008.
[10]  Zhou, Q., Li, K., Jun, X., and BO, L., Role and Functions of beneficial microorganisms in sustainable aquaculture. Bioresource Technology, 100: 3780-3786. 2009.
[11]  Son, V.M., Chang, C.C., Wu, M., Guu, Y., Chiu, C.H., and Cheng, W., Dietary administration of probiotic, Lactobacllus plantarum, enhanced the growth, innate immune responses and disease resistance of grouper Epinephelus coioides. Fish & Shellfish immunology, 26: 691-698. 2009
[12]  Agouz, H.M. and W., Effect of Biogen® and Myco-Ad® on the Growth Performance of Common Carp (Cyprinus carpio) Fed a Mycotoxin Contaminated Aquafeed. Journal of Fisheries and Aquatic Science, 6 (3): 334-345. 2011.
[13]  Wang, Y.C., Chang, P.S., and Chen, H.Y., Differential time series expression of immunerelated genes of Pacific white shrimp in response to dietary inclusion of â 1, 3-glucan. Fish & Shellfish Immunology, 24: 113-121. 2008.
[14]  Lakshmanan, R. and ., . Research Journal of Microbiology, 3 (3): 198-203. 2008.
[15]  Cutting, S.M., Bacillus probiotics. Food Microbiology. 28(2):214-220. 2010.
[16]  Farzanfar, A., Lashtooaghaee, Gh., Alizadeh, M., Bayati, M., and Ghorbani, R., Effect of using Bacillus subtilis and Bacillus licheniformis as probiotic on growth parameters, survival and carcass quality in rainbow trout, Oncorhynchus mykiss (Walbaum) fry. In: The first national coldwater fishes conference, Tonekabon-Iran, 12-14 May 2009. pp: 154. 2009.
[17]  NRC., Nutrient Requirement of Fish. National Academic Press, Washington, DC, USA.Pages:114. 1993.
[18]  Newaj-Fyzul, A., A.A., , A., , A., B., Bacillus subtilis AB1 controls Aeromonas infection in rainbow trout, Oncorhynchus mykiss (Walbaum). Applied Microbiology, 103: 1699-1706. 2007.
[19]  MacFarland, J.F., Biochemical testes for identification of medical bacteria. Williams and Wilkins. 912P. 2000.
[20]  Larsen, H.N., Comparison of various methods of hemoglobin detection of channel catfish blood. The Progressive Fish-Culturist, 26: 1-15. 1964.
[21]  Abdelhamid, A.M., Mehrim, A.I., El-Barbary, M.I., Ibrahim, S.M., and Abd El-Wahab, A.I., Evaluation of a New Egyptian Probiotic by African Catfish Fingerlings. Journal of Environmental Science and Technology, 2: 133-145. 2009.
[22]  IMBC,. Basic Techniques in Fish Haematology: A Guide to the Practical Skills in Collecting and Analysing Fish Blood Samples. Aqualex Multimedia Consortium, UK. 1998.
[23]  Benfey, T.J. and Sutterlin, A.M., The haematology of triploid landlocked Atlantic salmon, Salmo salar. Journal of Fish Biology, 24: 333-338. 1984.
[24]  Thrall, M.A., Veterinary Hematology and Clinical Chemistry ED., Lippincott Williams and Wilkins, Maryland, USA. 2004.
[25]  Yildirim, M., Lim, C., Wan, P., and Klesius, P.H., Growth performance and immune response of channel catfish (Ictalurus punctatus) fed diets containing graded levels of gossypol-acetic acid. Aquaculture, 219: 751-768. 2003.
[26]  Ellis, A.E., Lysozyme Assays: In Stolen Aal;SOS. Tech. Fish Immunology, 101-103. 1990.
[27]  Akhlaghi, M., and Keshavarz, M., Occurence of streptococcosis in rainbow trout, farms of Fars province. Iranian journal of veterinary research, 2: 183-189. 2002.
[28]  Trachoo, N., and Journal of Biological Sciences, 6 (1): 202-208. 2006.
[29]  Austin, B., Stuckey, L.F., Robertson, P.A.W., Effendi, J., and Griffith, D.R.W., A probiotic strain of Vibrio alginolyticus effective in reducing diseases caused by Aeromonas salmonicida, Vibrio anguillarum and Vibrio ordalii. Journal of Fish diseases, 18: 93-96. 1995.
[30]  Raida, M.K., Larsen, J.L., Nielsen, M.E., and Buchmann, K., Enhanced resistance of rainbow trout, Oncorhynchus mykiss (Walbaum), against Yersinia ruckeri challenge following oral administration of Bacillus subtilis and Bacillus licheniformis (BioPlus2B). Journal of Fish diseases, 26: 495-498. 2003.
[31]  Brunt, B., and Austin, B., Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of Fish Diseases, 28: 693-701. 2005.
[32]  De Carla Dias, D., de Stefani, M.V., Ferreira, C.M., Franca, F.M., Ranzani-Paiva, M.J.T., and Santos, A.A. Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics. , 41: 1064-1071. 2010.
[33]  Tavakoli, H., and Akhlaghi, M., Study of lysozyme, immunoglobulin, blood cell and hematocrit changes following experimental infection with a pathogenic Aeromonas hydrophila in rainbow trout. Iranian journal of veterinary research, 2: 157-162. 2009.
[34]  li, H.M., Ghazalah, A.A., and Gehad, E.A., Practical Aspects and Immune response of Probiotics Preparations Supplemented to Nile Tilapia (Oreochromis Niloticus) Diets. Nature and Science, 8: 39-45. 2010.
[35]  Sihag, R.C., and Sharma, P., Probiotics: The New Ecofriendly Alternative Measures of Disease Control for Sustainable Aquaculture. Journal of Fisheries and Aquatic Science, 7: 72-103. 2012.
[36]  Aly, S.M., Ahmed, Y.A.G., Ghareeb, A.A.A., and Mohamed, M.F., . Fish & Shellfish Immunology, 25: 128-136. 2008.
[37]  Vojgani, M., Immunology. Jahad Daneshgahi Publication. Iran, Tehran. 4:196. 2001.
[38]  Brunt, J., Newaj-Fyzul, A., and Austin, B., The development of probiotics for the control of multiple bacterial diseases of rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of Fish Diseases, 30: 573-579. 2007.
[39]  Sharifuzzaman, S.M., and Austin, B., Influence of probiotic feeding duration on disease resistance and immune parameters in rainbow trout. Fish & Shellfish Immunology, 27: 440-445. 2009.
[40]  Bandyopadhyay, P., and Das Mohapatra, P.K., Effect of a probiotic bacterium Bacillus circulans PB7 in the formulated diets: on growth, nutritional quality and immunity of Catla catla (Ham.). Fish Physiol Biochem, 35: 467-478. 2009.
[41]  Panigrahi, A., Kiron, V., Puangkaew, J., Kobayashi, T., Satoh, S., and Sugita, H., The viability of probiotic bacteria as a factor influencing the immune response in rainbow trout. Aquaculture, 243: 241-254. 2005.
[42]  Al-Dohail, M.A., Hashim, R., and Aliyu-Paiko, M., Effects of the probiotic, Lactobacillus acidophilus, on the growth performance, haematology parameters and immunoglobulin concentration in African Catfish (Clarias gariepinus, Burchell 1822) fingerling. , 40: 1642-1652. 2009.
[43]  Hoseinifar, S.H. and Pooramini, M., Application of probiotics and prebiotic in aquaculture. Green Wave Publication. Iran, Tehran. 2007.
[44]  Magnado´ ttir, B., Lange, S., Gudmundsdottir, S., Bøgwald, J., and Dalmo, R.A., Ontogeny of humoral immune parameters in fish. Fish & Shellfish Immunology, 19: 429-439. 2005.
[45]  Siwicki, A.K., Studnicka, M., Morand, M., Pozet, F., and Terech-Majewska, E., Comparative immunotoxicology-a new direction. Acta Vet (Brno), 67: 295-301. 1998.