Han Sun-Kyung, Shin Myeong-Su, Park Ho-Eun, Kim So-Young, Lee Wan-Kyu
College of Veterinary Medicine, Chungbuk National University, Cheongju 362-763, Korea.
Functional Food & Nutrition Division, Department of Agrofood Resource, National Academy of Agricultural Science, Rural Development Administration, Jeonju 560-500, Korea.
Korean J Food Sci Anim Resour. 2014;34(5):614-21. doi: 10.5851/kosfa.2014.34.5.614. Epub 2014 Oct 31.
This study was conducted to isolate and characterize bacteriocin-producing bacteria against Clostridium perfringens (C. perfringens) from domestic animals to determine their usefulness as probiotics. Bacteriocin-producing bacteria were isolated from pig feces by the spot-on-lawn method. A total of 1,370 bacterial stains were isolated, and six were tentatively selected after identifying the inhibitory activity against the pathogenic indicator C. perfringens KCTC 3269 and KCTC 5100. The selected strains were identified as Enterococcus faecalis (E. faecalis) by 16s rRNA sequencing. Most of the isolated bacterial strains were resistant to 0.5% bile salts for 48 h and remained viable after 2 h at pH 3.0. Some E. faecalis also showed strong inhibitory activity against Listeria monocytogenes KCTC 3569, KCTC 3586 and KCTC 3710. In the present study, we finally selected E. faecalis AP 216 and AP 45 strain based on probiotic selection criteria such as antimicrobial activity against C. perfringens and tolerance to acid and bile salts. The bacteriocins of E. faecalis AP 216 and AP 45 strains were highly thermostable, showing anticlostridial activities even after incubation at 121℃ for 15 min. These bacteriocinproducing bacteria and/or bacteriocins could be used in feed manufacturing as probiotics as an alternative to antibiotics in the livestock industry.
本研究旨在从家畜中分离并鉴定能产生抗产气荚膜梭菌细菌素的细菌,以确定其作为益生菌的效用。通过点种法从猪粪便中分离出能产生细菌素的细菌。共分离出1370株细菌菌株,在鉴定出对致病性指示菌产气荚膜梭菌KCTC 3269和KCTC 5100具有抑制活性后,初步筛选出6株。通过16s rRNA测序将筛选出的菌株鉴定为粪肠球菌。大多数分离出的细菌菌株在48小时内对0.5%的胆盐具有抗性,在pH 3.0条件下处理2小时后仍能存活。一些粪肠球菌对单核细胞增生李斯特菌KCTC 3569、KCTC 3586和KCTC 3710也表现出较强的抑制活性。在本研究中,我们最终根据对产气荚膜梭菌的抗菌活性以及对酸和胆盐的耐受性等益生菌选择标准,筛选出粪肠球菌AP 216和AP 45菌株。粪肠球菌AP 216和AP 45菌株的细菌素具有很高的热稳定性,即使在121℃下孵育15分钟后仍具有抗梭菌活性。这些产生细菌素的细菌和/或细菌素可作为益生菌用于饲料生产,以替代畜牧业中的抗生素。