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Mrvaljević I., Veljović K., Terzić-Vidojević A., Dinić M., Popović N., Golić N.
Invetlab Ltd., Adaševci, Serbia; Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Serbia

Exposure of cubes to environmental bacteria in the first days after birth is prerequisite for development of the immune system. Nevertheless, newborns could be infected by pathogens causing the intestinal infections accompanied with diarrhea and weight loss resulting in significant economic loss in the livestock production. Increased interest in use of probiotics in livestock has arisen from European regulations related to the use of antibiotics in livestock production. Since probiotics are considered as an appropriate alternative, number of studies deal with development of probiotics for animals based on use of lactic acid bacteria, Bacillus sp., or yeasts.

The strains Streptococcus thermophilus BGVLJ1-44, Lactobacillus fermentum BGHI14 and Lactobacillus helveticus BGRA43 were determined by nucleotide sequencing of 16S rDNA and pulse-field gel electrophoresis (PFGE). The main probiotic features recommended by EFSA were followed: susceptibility to antibiotics, antimicrobial activity, survival in simulated gastrointestinal tract (GIT) conditions, adhesion of the strains to Caco-2 epithelilal intestinal cells, adhesion of Escherichia coli ATCC25922 and Salmonella 654/7E (veterinary isolate) to Caco-2 cells in the presence and absence of probiotic strains, as well as in the presence of probiotic combination, and the effect of UV-inactivated strains, live strains and probiotic combination on GALT proliferation. The 25 gravid sows and 10 gravid goats treated with probiotic culture for 10 days before farrowing and 25 non-treated sows and 10 goats were followed. The general health of treated and control animals and the total number of LAB, Enterobacteriaceae, and Clostridium perfrigens in faeces was followed.

Tested strains succesfully survived in the simulated GIT conditions. The strains were sensitive to the recommended levels of antibiotics according to the EFSA recommendations. The strain BGRA43 exhibited broad spectrum of antimicrobial activity on Staphylococcus sp. (clinical isolate), S. aureus ATCC25923, Yersinia enterocolitica O3 (clinical isolate), Shigella sonnei (Hamburg IO5), Sh. flexneri (London 9950), Sh. dysenteriae (serotype 5 and 7), Streptococcus pneumoniae ATCC276, St. pneumoniae ATCC496, Pseudomonas sp., E. coli C600, E. coli ATCC25922 while probiotic combination shows a synergistic effect and exhibits the strongest antimicrobial effect on C. sporogenes, C. perfringens (veterinary isolate) and C. difficile (clinical isolate). The strain BGRA43 exhibited the highest level of adhesion to Caco-2 cells and significantly reduced the number of attached E. coli ATCC25922 (p<0.5). The probiotic combination exerts a synergistic effect and a significantly (p <0.1) reduced the number of attached E. coli ATCC25922. The strains BGVLJ1-44 (p<0.1) and BGRA43 (p<0.5) significantly reduce the number of attached Salmonella 654/7E. The live strains significantly increase the GALT lymphocytes proliferation. Besides, results of farm trial revealed reduction of Enterobacteriaceae in neonatal piglets after the probiotic treatment of pregnant sows, as well as reduction of C. perfringens in goats (below the detection threshold) after the probiotic treatment.

A novel improved probiotic culture in treatment of intestinal infections in animals was developed. The novel probiotic culture represents an alternative to use of antibiotics in treatment of intestinal infections, based on wide range of antimicrobial activity, specifically for severe hardly curable infections caused by C. perfrigens and C. difficille.

Keywords: Probiotic, Antimicrobial activity, Intestinal infections, Streptococcus thermophilus, Lactobacillus fermentum, Lactobacillus helveticus

Mrvaljević I., et al. (2016). Development of novel probiotic for prevention and treatment of intestinal infections in animals.Conference Proceedings of IPC2016. Paper presented at the International Scientific Conference on Probiotics and Prebiotics, Budapest (p. 108.). IPC2016

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