Isolation, Identification and Selection of Lactic Acid Bacteria from the Digestive System of Sistani Cattle for Probiotic Production

Authors

1 Student

2 Iran Scientific and Industrial Research Organization

3 Razi Serum Institute of Tehran

4 Faculty member of Zabol University

5 Faculty member of Atonos University

Abstract

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.

Keywords

References

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Journal of Ruminant Research
Volume 10, Issue 1
June 2022
Pages 119-133

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