نوع مقاله : مقاله پژوهشی
نویسندگان
1 دانشجوی دکتری، گروه علوم دامی، دانشکده کشاورزی، دانشگاه زابل، زابل، ایران
2 دانشیار ، گروه علوم دامی، دانشکده کشاورزی، دانشگاه زابل، ایران
3 دانشیار ، گروه دام، طیور و آبزیان، پژوهشکده کشاورزی، سازمان پژوهش های علمی و صنعتی ایران
4 استاد، موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، آموزش و ترویج کشاورزی، ایران
5 عضو هیات علمی دانشگاه ملی اتونوس مکزیک
چکیده
کلیدواژه‌ها
عنوان مقاله [English]
نویسندگان [English]
Abstract
Background: The bovine rumen is the suitable growth medium to many of the bacteria responsible for converting food into energy. In recent years, many rumen microbes have been identified and isolated using 16sRNA gene sequencing. Some microbes are suggested as food additives to promote the growth and production of animals. The use of probiotics as feed additive has been induced by stimulating the positive effects on the host by creating a favorable balance of intestinal microflora. Therefore, is study aimed to isolate, identify and select lactic acid bacteria to produce probiotics from the rumen and feces of Sistani cows.
Material and Method: In this experiment, samples were taken from the rumen of 12 and the feces of 4 Sistani cows in the industrial slaughterhouse of Zabol city. The samples were transferred to test tubes containing liquid MRS culture medium and incubated at 39 ° C for 48 hours. After growing from the mentioned medium, the plates containing MRS agar were linearly cultured and the plates were incubated in aerobic conditions at 39 ° C for 48 hours. The emerged colonies were examined for growth characteristics and morphology as well as purity. Gram-positive, spore-free, spherical negative catalase, and rod-shaped bacteria that did not produce beta hemolysis were tested for antimicrobial, acid, bile salt, and antibiotic susceptibility testing.
Results: Of the initial 158 isolates of lactic acid bacteria, 84 isolates were tested for antimicrobial activity against four species of Salmonella and one species of E. coli. Then the isolates obtained from the antimicrobial test were examined for pH resistance test. Thirteen samples selected from this experiment were studied for bile resistance test, of which five isolates were examined for antibiotic susceptibility testing and entered the final stage for genetic and molecular testing.There were only two isolates that were obtained by molecular testing and after sequencing and using NCBI software had a high genetic affinity with two bacteria Streptoccos infantarius (isolate A) and Entrococcos faecium isolate B. These two isolates were observed as potential probiotics in Sistani cows.
Conclusion: The general results of the study showed that isolates A and B have suitable properties for the use of probiotics. . These characteristics are high survival rate at low pH and 3% bile concentration and inhibition of pathogenic bacteria such as E. coli and Salmonella, as well as antibiotic resistance. Therefore, isolates A and B will later be used as probiotics the ruminants feed.
کلیدواژه‌ها [English]
9.Fuller, R. 1989. Probiotic in man and animals. Journal of Applied Bacteriology, 66: 365-378.
10.Gill, H.S., Shu, Q., Lin, H., Rutherfurd, K. J. and Cross, M. L. 2001. Protection against translocating Salmonella typhimurium infection in mice by feeding the immuno-enhancing probiotic Lactobacillus rhamnosus strain HN001. Medical Microbiology Immunology, 190: 97–104.
11.Gilliland, S.E., Staley, T.E. and Bush, L.J. 1984. Importance of bile tolerance of Lactobacillus acidophilus used as a dietary adjunct. Journal of Dairy Science. 67: 3045- 3051.
12.Gilliland, S.E. 1990. Health and nutritional benefits from lactic acid bacteria. Federation of European Microbiological Societies, 7: 175- 186.
13.Gotcheva, V., Histozova, E., Hristozova, T., Guo, M., Roshkova, Z. 2007. Assessment of potential probiotic properties of lactic acid bacteria and yeast strains. Food Biotechnology, 16: 211-225.
14.Goldin, B. and Gorbach, S. 1992. Probiotics for humans. Londron: Chapman and Hall, 355-376.
15.Havenaar, R., Brink, B.T., Huis in,t Veld, J.H.J. 1992. Selection of strains for probiotic use. Chapman and Hall, London, 209–224.
16.Hood, S.K. and Zotolla, E.A. 1998. Effect of low pH on viability of Lactobacillus acidophilus to survive and adhere to human intestinal cells. Journal of Food Science, 53:1514-1516.
17.ISO-15214. 1998. Microbiology of food animal breeding stuffs-Horizontal method for the enumeration of Mesophilic Lactic Acid Bacteria-colony count technique 15214.
18.Jin, L.Z., Ho, Y.W., Abdullah, M.A., Ali, M.A. and Jalaludin, S. 1996. Adhesion of intestinal Lactobacillus isolates to intestinal epithelial cells of chicken. Letters in Applied Microbiology, 22:229-232.
19.Konstantinov, S.R., Favier, C.F., Zhu, W.Y., Williams, B.A., Klu_, J., Souffrant, W.-B., De Vos, W.M., Akkermans, A.D.L. and Smidt, H. 2004. Microbial diversity studies of the porcine gastrointestinal ecosystem during weaning transition. Animal Research, 53: 317–324.
20.Lozupone, C., Faust, K., Raes, J., Faith, J.J., Frank, D.N., Zaneveld, J., Gordon, J.I. and Knight, R. 2012. Identifying genomic and metabolic features that can underlie early successional and opportunistic lifestyles of human gut symbionts. Genome Research, 22: 1974–1984.
21.Madureira, A.R., Pereira, C.I., Truszkowska, K., Gomes, A.M., Pintado, M.E. and Malcata, F.X. 2005. Survival of probiotic bacteria in a whey cheese vector submitted to environmental conditions prevailing in the gastrointestinal tract. International of Dairy Journal, 15: 921-927.
22.Makras, L., De Vuyst, L. 2006. The in vitro inhibition of Gram-negative pathogenic bacteria by bifidobacteria is caused by the production of organic acids. International of Dairy Journal, 16: 1049–1057.
23.Maragkoudakis, P.A., Mountzouris, K.C., Psyrras, D., Cremonese, S., Fischer, J., Canter, M.D. and Tsakalidou, E. 2009. Functional properties of novel protective lactic acid bacteria and application in raw chicken meat against Listeria monocytogenes and Salmonella enteridis. International Journal of Food Microbiology, 3: 219-226.
24.Mathur, S. and Singh, R. 2005. Antibiotic resistance in food lactic acid bacteria. International Journal Food Microbiology, 105: 281-295.
26.Meyer, P.M., Vaz Pires, A., Bagaldo, A.R., Correira de Simas, J.M. and Susin, I. 2001. Addition of probiotic to whole milk or milk replacer and Holstein calves’ performance. Scientia Agricology, 58: 215–221.
27.Musikasang, H. 2008. Screening of lactic acid bacteria as probiotics in chicken and enhancement of bacteria survival by microencapsulation. Master Thesis, Prince of Songkla University, Thailand.
28.Olejnik, A., Lewandowska. M., Obarska, M. and Grajek, W. 2005. Tolerance of Lactobacillus and Bifidobacterium strains to low pH, bile salts and digestive enzymes. Electronic Journal of Polish Agricultural Universities, 8: 25-32.
29.Ouwehand, A.C., Kirjavainen, P.V., Shortt C. and Salminen, S. 1999. Probiotics: mechanisms and established effects. Iranian Journal of Pharmaceutical Research. 9: 43-52.
30.Oyarzabal, A.O. and D.E. Conner. 1995. In vitro fructooligosaccharide utilization and inhibition of Samonella spp. by selected bacteria. Poultry Science, 74: 1418-1425.
31.Ponce, A.G., Moreira, M.R., Valle, C.E. and Roura S.I. 2008. Preliminary characterization of bacteriocinlike substances from lactic acid bacteria isolated from organic leafy vegetables. Food Science and Technology, (41)3: 432-441.
32.Seeley, H.W. Jr. and VanDemark, P.J. 1981. Microbes in action. New Biological books, 39: 429-440.
33.Shehata, M.G., El Sohaimy, S.A., El-Sahn , M. A. and Youssef, M.M. 2016. Screening of isolated potential probiotic lactic acid bacteria for cholesterol lowering property and bile salt hydrolase activity. Food Science and Technology Department, 61: 65-75.
34.Singleton, P. 1999. Bacteria in biology, biotechnology and medicine. 5th ed. John Wiley, UK.
35.Sornplang, P., 2009. Characterization of Lactobacillus from Thai native chicken gut and utilization in the diet to improve growth performance and cholesterol reduction in broilers. Ph.D. Thesis, Khon Kaen University, Thailand.
36.Suskovic,J., Blazenka, K., Jasna B., Andreja, L., Ksenija, H. and Srecko, M. 2010. Antimicrobial Activity of Lactic Acid Bacteria, Food Technology of Biotechnology, 48 (3): 296–307.
37.Tambekar, D.H. and Bhutada, S.A. 2010. Studies on antimicrobial activity and characteristics of Bacteriocins produced by Lactobacillus strains isolated from milk of domestic animals. Internet Journal of Microbial, 8: 1-6.
38.Timmerman, H. M., Mulder, L., Everts, H., van Espen, D.C., van der Wal, E., Klaassen, G., Rouwers, S.M.G., Hartemink, R., Rombouts, F.M. and Beynen, A.C. 2005. Health and growth of veal calves fed milk replacers with or without probiotics. Journal of Dairy Science, 88: 2154–2165
39.Treagan, L. and Pulliam, L. 1982. Medical Microbiology Laboratory Procedures. W.B. Saunders Company, Philadelphia, pp 233-243.
40.Xanthopoulos, V., Litopoulou-Tzanetaki, E. and Tzanetakis, N. 1997. Lactic Acid Bacteria. Caen Presses Universitaires de Canada. Wayne, Pa.
41.Wen Hsin, L., Bi Yu., Sheng Hon, H. and Hau Yang T. 2007. Different probiotic properties for Lactobacillus fermentum strains isolated from swine and poultry. Journal Anaerobe, 107-113.
42.Wheeler, W.E. and Noller, C.H. 1997. Gastrointestinal tract pH and starch in feces of ruminants. Journal of Animal Science, 44: 131-135.
43.Yu, B. and Tsen, H.Y. 1993. Lactobacillus cells in the rabbit digestive tract and the factors affecting their distribution. Journal Applied Bacteriology, 75: 269-275.
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