毛皮动物源粪肠球菌的分离鉴定

doi:10.16431/j.cnki.1671-7236.2020.09.035

摘要/Abstract

摘要： 为获得毛皮动物源粪肠球菌，并评价其益生特性，本研究从健康成年毛皮动物（水貂、狐狸、貉）粪便中分离粪肠球菌，通过形态学观察、生化试验和16S rRNA序列分析等方法进行种属鉴定。测定分离菌株生长曲线、产酸能力、抗菌药敏感性，并挑取部分菌株测定其对于温度、人工胃液、人工胆盐的耐受能力。结果表明，分离菌株中共5株为革兰氏阳性菌，生化特性与粪肠球菌标准株基本相符，且经16S rRNA序列分析鉴定为粪肠球菌。5株菌均于培养后2 h进入对数期，8~10 h进入稳定期，且具有弱产酸能力。分离菌株对四环素、左氟沙星耐药性强，耐药率为100%，其次为青霉素（80%）、红霉素（80%）、庆大霉素（80%）、氯霉素（40%），对氨苄西林和万古霉素敏感。水貂、狐狸和貉源的粪肠球菌对于60 ℃以下温度处理、pH>3.0的人工胃液、0.3%~0.5%浓度的胆盐耐受能力较强，对70 ℃以上高温和pH<3.0的人工胃液耐受性差。综上所述，本研究共获得5株毛皮动物源（水貂、狐狸、貉）粪肠球菌，分离菌株繁殖较快，适合在毛皮动物肠道中定植并发挥益生作用，且具有较好的益生特性和抗逆性，可作为动物用微生态制剂的候选菌种进一步研究。

关键词: 毛皮动物; 粪肠球菌; 生长曲线; 耐药性; 稳定性

Abstract: In order to obtain Enterococcus faecalis from fur animals and evaluate its prebiotic properties,in this study,Enterococcus faecalis was isolated from the feces of healthy adult fur-bearing animals (mink,fox,raccoon dog),identified by morphological observation,biochemical test and 16S rRNA sequence analysis.The growth curve,acid production capacity and antibiotic sensitivity of the Enterococcus faecalis isolates were measured to evaluate their probiotic properties.Some strains were selected to determine their tolerance to temperature,artificial gastric juice and artificial bile salt.The results showed that five strains were Gram-positive,and their biochemical characteristics were basically consistent with the standard strains of Enterococcus faecalis,and they were identified as Enterococcus faecalis by 16S rRNA sequence analysis.The five strains all entered the logarithmic phase at 2 h after culture,and entered the stable phase at 8-10 h,and had weak acid production capacity.The resistance rate of the isolates to tetracycline and levofloxacin was 100%,followed by penicillin (80%),erythromycin (80%),gentamicin (80%) and chloramphenicol (40%).All the isolates were sensitive to ampicillin and vancomycin.Enterococcus faecalis from mink,fox and raccoon dog had strong tolerance to temperature below 60 ℃,artificial gastric juice with pH>3.0 and 0.3%-0.5% concentration of bile salt,but poor tolerance to temperature above 70 ℃,and artificial gastric juice with pH<3.0.In conclusion,five strains of Enterococcus faecalis from fur animals (mink,fox,raccoon dog) were obtained in this study.The isolated strains propagated rapidly,which were suitable for colonization and played a prebiotic role in fur animals' intestines,and had good prebiotic characteristics and stress resistance.They could be used as candidate strains for animal microbiological agents for further study.

Key words: fur-bearing animals; Enterococcus faecalis; growth curve; drug resistance; stability

中图分类号:

S858.92

慈洋, 史晓洁, 宋鹏, 丁琦, 李文立. 毛皮动物源粪肠球菌的分离鉴定[J]. 中国畜牧兽医, 2020, 47(9): 2997-3005.

CI Yang, SHI Xiaojie, SONG Peng, DING Qi, LI Wenli. Isolation and Identification of Enterococcus faecalis from Fur-bearing Animals[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2997-3005.

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