In vitro probiotic properties of Bacillus sp isolated from biofloc reared genetically improved farmed tilapia (Oreochromis niloticus)

M MENAGA; S FELIX; M CHARULATHA

doi:10.56093/ijans.v89i5.90034

Authors

M MENAGA PhD Scholar, Advanced Research Farm Facility Dr MGR Fisheries College and Research Institute, Chennai, Tamil Nadu.
S FELIX Vice-Chancellor, Tamil Nadu Dr J. Jayalalithaa Fisheries University, Nagapattinam, Tamil Nadu 611 012 India
M CHARULATHA Senior Research Fellow, Dr MGR Fisheries College and Research Institute, Ponneri, Tamil Nadu

https://doi.org/10.56093/ijans.v89i5.90034

Keywords:

Antimicrobial activity, Biofloc, GIFT, In vitro properties, Probiotics

Abstract

A study was conducted to investigate the probiotic potential of the bacterial species from biofloc reared genetically improved farmed tilapia (GIFT) using in vitro quantitative assays. Based on the morphological, biochemical and 16S rRNA sequencing analysis, isolated bacterial species from GIFT gut were identified as Bacillus infantis, Exiguobacterium profundum, Bacillus subtilis and Bacillus megaterium. The in vitro probiotic properties such as bile salt hydrolase activity, bile tolerance, growth at different bile concentrations, antibiotic susceptibility test, antimicrobial activity, auto aggregation test, bacterial adhesion to hydrocarbons and resistance to gastric acidity were performed. All the isolates showed higher resistance to bile tolerance test and growth of cultures was observed from 0.5 to 8% bile salt concentrations. The distinct zone of hydrolysis was marked in the tested isolates in bile salt hydrolysis activity. Except B. infantis, all the other three isolates were predominantly resistant to the tested antibiotics. Antimicrobial activity against three pathogens, viz. Vibrio parahaemolyticus, Vibrio harveyi and Aeromonas hydrophila was observed in all the isolates. E. profundum and B. subtilis showed improved auto aggregation. Enhanced resistance to bile salt adhesion to hydrocarbon and in vitro gastric acidity (pH 3) was seen in B. megaterium. It is one of such unique studies confirming the probiotic effect of Bacillus sp. isolated mainly from GIFT biofloc culture. B. subtilis and B. megaterium exhibited remarkable in vitro probiotic properties and thus can be recommended as a successful probiotic strain for fish farming.

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References

Aly S M, Abd-El-Rahman A M, John G and Mohamed M F. 2008. Characterization of some bacteria isolated from Oreochromis niloticus and their potential use as probiotics. Aquaculture. 277: 1–6. DOI: https://doi.org/10.1016/j.aquaculture.2008.02.021

Begley M, Hill C and Gahan C G M. 2006. Bile salt hydrolase activity in probiotics. Applied and Environmental Microbiology 72: 1729–38. DOI: https://doi.org/10.1128/AEM.72.3.1729-1738.2006

Brashears M M, Galyean M L, Loneragan G H and Killinger- Mann K. 2003. Prevalence of Escherichia coli O157, H7 and performance by beef feed lot cattle given Lactobacillus direct- fed microbials. Journal of Food Protection 66: 748–54. DOI: https://doi.org/10.4315/0362-028X-66.5.748

Brown J H. 1939. Bergey’s Manual of Determinative Bacteriology. Canzi E, Guglielmetti S, Mora D, Tamagnini I and Parini C. 2005. Conditions affecting cell surface properties of human intestinal Bifidobacteria. Antonie van Leeuwenhoek 88: 207–19 DOI: https://doi.org/10.1007/s10482-005-6501-3

Chateau N, Deschamps A M and Hadj-Sassi A. 1994. Heterogeneity of bile salts resistance in the Lactobacillus isolates from a probiotic consortium. Letters in Applied Microbiology 18: 42–44. DOI: https://doi.org/10.1111/j.1472-765X.1994.tb00796.x

Chaurasia B, Pandey A, Palni L M S, Trivedi P, Kumar B and Colvin N. 2005. Diffusible and volatile compounds produced by an antagonistic Bacillus subtilis strain cause structural deformations in pathogenic fungi in vitro. Microbiological Research 160(1): 75–81. DOI: https://doi.org/10.1016/j.micres.2004.09.013

Clinical and Laboratory Standards Institute. 2016. Performance standards for antimicrobial susceptibility testing; 21st informational supplement. CLSI document M100-S21. Clinical and Laboratory Standards Institute, Wayne, PA.

Costa-Pierce B A. 2010. Sustainable ecological aquaculture systems: The need for a new social contract for aquaculture development. Marine Technology Society Journal 44: 88–112. DOI: https://doi.org/10.4031/MTSJ.44.3.3

Defoirdt T, Halet D, Vervaeren H, Boon N, Van de Wiele T, Sorgeloos P, Bossier P and Verstraete W. 2007. The bacterial storage compound poly--hydroxybutyrate protects Artemia franciscana from pathogenic Vibrio campbelli. Environmental Microbiology 9(2): 445–52. DOI: https://doi.org/10.1111/j.1462-2920.2006.01161.x

Dinh T N, Wille M, De Schryver P, Defoirdt T, Bossier P and Sorgeloos P. 2010. The effect of poly--hydroxybutyrate on larviculture of the giant freshwater prawn (Macrobrachium rosenbergii). Aquaculture 302: 76–81. DOI: https://doi.org/10.1016/j.aquaculture.2010.02.011

Drablos F, Nicholson D and Ronning M. 1999. EXAFS study of zinc coordination in Bacitracin A. Biochimica et Biophysica Acta 1431: 433–42. DOI: https://doi.org/10.1016/S0167-4838(99)00064-3

EFSA (European Food Safety Authority). 2008. Update of the criteria used in the assessment of bacterial resistance to antibiotics of human or veterinary importance. EFSA Journal 32: 1–15.

FAO. 2016. The State of World Fisheries and Aquaculture (SOFIA): Contributing to food security and nutrition for all. Rome, 2016. 200 p. Food and Agriculture Organization (FAO), Rome.

Felix S, Antony Jesu Prabhu P, Cheryl Antony, Gopalakannan A and Rajaram R. 2015. Studies on nursery rearing of Genetically Improved Farmed Tilapia (GIFT) in biosecured raceway systems in India. Journal of Indian Fisheries Association 42: 1–10.

Gullian M, Thompson F and Rodríguez J. 2004. Selection of probiotic bacteria and study of their immunostimulatory effect in Penaeus vannamei. Aquaculture 233: 1–14. DOI: https://doi.org/10.1016/j.aquaculture.2003.09.013

Halet D, Defoirdt T, Van Damme P, Vervaeren H, Forrez I, Van de Wiele T, Boon N, Sorgeloos P, Bossier P and Verstraete W. 2007. Poly--hydroxybutyrate-accumulating bacteria protect gnobiotic Artemia franciscana from pathogenic Vibrio campbelli. FEMS Microbiology Ecology 60: 363–69. DOI: https://doi.org/10.1111/j.1574-6941.2007.00305.x

Hong A H, Duc H L and Cutting S M. 2004. Intracellular fate and immunogenicity of Bacillus subtilis spores. Vaccine 22: 1873– 85. DOI: https://doi.org/10.1016/j.vaccine.2003.11.021

Hyronimus B, Le Marrec C, Hadj Sassi A and Deschamps A. 2000. Acid and bile tolerance of spore-forming lactic acid bacteria. International Journal of Food Microbiology 61: 193– 97. DOI: https://doi.org/10.1016/S0168-1605(00)00366-4

Iyapparaj P, Maruthiah T, Ramasubburayan R, Prakash S, Kumar C, Immanuel G and Palavesam A. 2013. Optimization of bacteriocin production by Lactobacillus sp. MSU3IR against shrimp bacterial pathogens. Aquatic Biosystems 9(1): 12. DOI: https://doi.org/10.1186/2046-9063-9-12

Johansen R, Needham J R, Colquhoun D J, Poppe T T and Smith A J. 2006. Guidelines for health and welfare monitoring of fish used in research. Laboratory Animals 40: 323–40. DOI: https://doi.org/10.1258/002367706778476451

Liu H, Li Z, Tan B, Lao Y, Duan Z, Sun W and Dong X. 2014. Isolation of a putative probiotic strain S12 and its effect on growth performance, non-specific immunity and disease- resistance of white shrimp, Litopenaeus vannamei. Fish and Shellfish Immunology 41: 300–07. DOI: https://doi.org/10.1016/j.fsi.2014.08.028

Luis-Villaseñor I E, Macías-Rodríguez M E, Gómez-Gil B, Ascencio-Valle F and Campa-Córdova I. 2011. Beneficial effects of four Bacillus strains on the larval cultivation of Litopenaeus vannamei. Aquaculture 321(1–2): 136–44. DOI: https://doi.org/10.1016/j.aquaculture.2011.08.036

Morikawa M, Ito M and Imanaka T. 1992. Isolation of a new surfactin producer Bacillus pumilus A-1, and cloning and nucleotide sequence of the regulator gene, psf-1. Journal of Fermentation and Bioengineering 74: 255–61. DOI: https://doi.org/10.1016/0922-338X(92)90055-Y

Nithya V and Halami P M. 2013. Evaluation of the probiotic characteristics of Bacillus species isolated from different food sources. Annals of Microbiology 63(1): 129–137. DOI: https://doi.org/10.1007/s13213-012-0453-4

Otero M C, Ocana V S and Nader-Macias E M. 2004. Bacterial surface characteristics applied to selection of probiotic microorganisms. Methods in Molecular Biology 268: 435–40.

Perez C, Suarez C and Castro G R. 1993. Antimicrobial activity determined in strains of Bacillus circulans cluster. Folia Microbiologica 38: 25–28. DOI: https://doi.org/10.1007/BF02814544

Pinchuk I V, Bressollier P, Verneuil B, Fenet B, Sorokulova I B, Megraud F and Urdaci M C. 2001. In vitro anti-Helicobacter pylori activity of the probiotic strain Bacillus subtilis is due to secretion of antibiotics. Antimicrobial Agents Chemotherapy 45: 3156–61. DOI: https://doi.org/10.1128/AAC.45.11.3156-3161.2001

Sorokulova I B, Pinchuk I V, Denayrolles M, Osipova I G, Huang J M, Cutting S M and Urdaci M C. 2008. The safety of two Bacillus probiotic strains for human use. Digestive Disease and Sciences 53: 954–63. DOI: https://doi.org/10.1007/s10620-007-9959-1

Sinha A K, Baruah K and Bossier P. 2008. Horizon Scanning: the potential use of biofloc as an anti-infective strategy in aquaculture—An overview. Aquaculture Health International 13: 8–10.

Sieladie D, Zambou N, Kaktcham P, Cresci A and Fontech F. 2011. Probiotic properties of Lactobacilli strains isolated from raw cow milk in the western highlands of Cameroon. Innovative Romanian Food Biotechnology 9: 12–28.

Shruthy V, Gowry P and Ghosh A. 2011. Probiotic potentials among Lactic acid bacteria isolated from curd. International Journal of Research in Ayurveda and Pharmacy 2(2): 602– 09.

Tambekar D and Bhutada S. 2010. An evaluation of probiotic potential of Lactobacillus sp. from milk of domestic animals and commercial available probiotic preparations in prevention of enteric bacterial infections. Recent Research in Science and Technology 2(10): 82–88.

Telli G S, Ranzani-Paiva M J T, Dias D C, Sussel F R, Ishikawa C M and Tachibana L. 2014. Dietary administration of Bacillus subtilis on hematology and non-specific immunity of nile tilapia (Oreochromis niloticus) raised at different stocking densities. Fish and Shellfish Immunology 39: 305–11. DOI: https://doi.org/10.1016/j.fsi.2014.05.025

Verdegem M C J. 2013. Nutrient discharge from aquaculture operations in function of system design and production environment. Reviews in Aquaculture 5: 158–71. DOI: https://doi.org/10.1111/raq.12011

Watanabe W O, Losordo T M, Fitzsimmons K and Hanley F. 2002. Tilapia production systems in the Americas: technological advances, trends, and challenges. Reviews in Fisheries Science 10: 465–98. DOI: https://doi.org/10.1080/20026491051758

Yilmaz M, Soran H and Beyatli Y. 2006. Antimicrobial activities of some Bacillus spp. strains isolated from the soil. Microbiological Research 161: 127–31. DOI: https://doi.org/10.1016/j.micres.2005.07.001