Department of Veterinary Medicine, College of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan.
Graduate Institute of Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan.
Arch Microbiol. 2020 Sep;202(7):1849-1860. doi: 10.1007/s00203-020-01908-w. Epub 2020 May 23.
The occurrence of multidrug-resistant pathogenic bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA), multidrug-resistant Acinetobacter baumannii (MDRAB), extended-spectrum β-lactamase (ESBL) Escherichia coli, and Pseudomonas aeruginosa, has become a serious problem in animals and public. The objective of this study was to identify and isolate lactic acid bacterial (LAB) strains from the intestinal tracts of pigs and feces of dogs and then characterize them as potential probiotics with antimicrobial activity against multidrug-resistant pathogenic bacteria. In a preliminary isolation screening, 45 of 1167 isolated LAB strains were found to have anti-S. aureus ATCC 27,735 activity. Using 16S rDNA and 16S-23S rDNA intergenic spacer region (ISR) sequences, five of these isolates were further identified as Lactobacillus animalis 30a-2, Lactobacillus reuteri 4-12E, Weissella cibaria C34, Lactococcus lactis 5-12H, and Lactococcus lactis 6-3H. Antimicrobial substance assays suggest that the L. lactis 5-12H, L. lactis 6-3H, L. animalis 30a-2, L. reuteri 4-12E, and W. cibaria C34 strains might produce bacteriocins and hydrogen peroxide (HO) as antimicrobial substances. The L. animalis 30a-2 and W. cibaria C34 strains were further characterized for probiotic properties and shown to have high acid and bile salt tolerance. Additionally, they have broad antimicrobial spectra, and can significantly repress the growth of all of the tested strains of MRSA isolates, some MDRAB, ESBL E. coli, and P. aeruginosa isolates, along with food-borne pathogenic bacteria such as Bacillus cereus ATCC 11778, Listeria monocytogens ATCC 19111, Salmonella spp., Shigella spp., and Yersinia enterocolitica BCRC 12986. This is the first report of HO-producing L. animalis 30a-2 and W. cibaria C34 isolated from the intestinal tracts of pigs and feces of dogs that have good antimicrobial activity against multidrug-resistant and food-borne pathogenic bacteria and have excellent probiotic properties.
耐多药病原菌(如耐甲氧西林金黄色葡萄球菌(MRSA)、多药耐药鲍曼不动杆菌(MDRAB)、超广谱β-内酰胺酶(ESBL)大肠杆菌和铜绿假单胞菌)的出现已成为动物和公众的严重问题。本研究的目的是从猪肠道和犬粪便中分离和鉴定乳酸菌(LAB)菌株,然后将其鉴定为具有抗多药耐药病原菌活性的潜在益生菌。在初步分离筛选中,从 1167 株分离的 LAB 菌株中发现有 45 株对金黄色葡萄球菌 ATCC 27735 具有活性。通过 16S rDNA 和 16S-23S rDNA 基因间隔区(ISR)序列,进一步鉴定其中 5 株为动物乳杆菌 30a-2、雷氏乳杆菌 4-12E、魏斯氏菌 C34、乳球菌 5-12H 和乳球菌 6-3H。抗菌物质检测表明,L. lactis 5-12H、L. lactis 6-3H、L. animalis 30a-2、L. reuteri 4-12E 和 W. cibaria C34 菌株可能产生细菌素和过氧化氢(HO)作为抗菌物质。进一步对动物乳杆菌 30a-2 和魏斯氏菌 C34 菌株进行了益生菌特性的表征,发现它们具有较高的酸和胆盐耐受性。此外,它们具有广谱的抗菌谱,可以显著抑制所有测试的耐甲氧西林金黄色葡萄球菌分离株、部分多药耐药鲍曼不动杆菌、超广谱β-内酰胺酶大肠杆菌和铜绿假单胞菌分离株以及食源性病原菌如蜡样芽胞杆菌 ATCC 11778、单核细胞增生李斯特菌 ATCC 19111、沙门氏菌、志贺氏菌和肠出血性大肠杆菌 BCRC 12986 的生长。这是首次报道从猪肠道和犬粪便中分离出具有良好的抗多药耐药和食源性病原菌活性的产 HO 动物乳杆菌 30a-2 和魏斯氏菌 C34,并且具有优良的益生菌特性。