Bacteriocin and new bacterial isolates
A bacteriocin and bacteria technology, applied in the fields of new bacteriocin, diseases in poultry, therapeutic compositions, and lactic acid-producing bacterial strains, and can solve the problems of difficult regulation of immune response, etc.
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Embodiment 1
[0082] Two new antagonistic strains producing bacteriocins, Lactobacillus salivarius, designated NRRL B-30514 (strain PVD32) and Lactobacillus acidophilus, designated NRRL B-30510 (strain LWP320) were isolated from the cecum and crop of broiler chickens come out. The cecum and crop were pre-emptied and washed twice with approximately 100 ml of sterile 0.85% w / v saline solution (physiological saline). The cecum and crop were suspended in sterile saline solution at a pH of approximately 7.0. Approximately 0.1 ml of the 10-fold diluted suspension was plated directly onto selective MRS agar. Plates were incubated anaerobically at approximately 37°C for approximately 16-18 hours. At least 10 Lactobacillus-specific colonies were selected from each sample. Lactobacilli were identified using the APO 50CHL micro detection system (bioMerieux, France).
[0083]The strains L. salivarius PVD32 and L. acidophilus LWP320 were grown on MRS agar at about 37° C. for about 24 hours. These t...
Embodiment 2
[0099] Two novel antagonistic bacteriocin-producing species, Enterococcus faecalis 21 (NRRL B-30645), and Enterococcus tenus 26 (NRRL B-30511), were isolated from cecum and crop smears of healthy broilers. The cecum and crop were emptied and washed twice with sterile 0.085% w / v saline solution (physiological saline) at approximately pH 7.0. The cecum and crop were suspended in sterile saline at approximately pH 7.0. About 0.1 ml of the 10-fold diluted suspension was plated directly on MRS selective medium and the plates were incubated at about 37°C for about 24-48 hours. Enterococci were identified using the micro-detection kit EN-COCCUS.
[0100] Campylobacter jejuni ATCC 11168 was used as a test culture to evaluate the antagonistic activity of the isolates, as described in Example 1 above. Thirty-three other species were used as test cultures to evaluate the antagonistic activity of the isolates, as described in Example 1 above. The antagonistic activity of the isolates a...
Embodiment 3
[0103] Crude antibacterial preparations were prepared by ammonium sulfate precipitation only of cultures of antagonistic strains of Lactobacillus grown under aerobic conditions at 37°C for approximately 18 hours on starvation medium:
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[0106] Add distilled water to 1000ml, the pH value is about 7.2
[0107] The culture fluid was centrifuged at approximately 12,000 xg for approximately 10 minutes. 1N NaOH was added to the obtained supernatant to adjust the pH to about 6.2, and about 130 U / ml catalase was added to remove organic acids, hydrogen peroxide, and inhibitory factors. Antagonist peptides were isolated from the supernatant by a combination of ammonium sulfate precipitation, desalting chromatography and gel filtration to obtain a crude preparation (CAP). CAP samples were filtered through a 0.22[mu] filter (Millipore, Bedford, MA).
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