阿合奇县猎鹰粪源海氏肠球菌的分离鉴定及其生物学特性分析

doi:10.16431/j.cnki.1671-7236.2024.10.039

摘要/Abstract

摘要： 【目的】自健康猎鹰粪便中分离具有益生作用的乳酸菌，为益生菌制剂的开发研究提供候选菌株。【方法】自阿合奇县采集6份猎鹰粪便样本，采用选择性培养基进行细菌分离培养，通过革兰染色、生化鉴定和16S rDNA PCR扩增和序列分析对分离菌进行鉴定，并对其进行生长曲线、产酸曲线、抗逆性、药物敏感性、体外抑菌、自聚集能力和表面疏水性等生物学特性测定和动物安全性试验。【结果】自猎鹰粪便中分离出3株分离菌，经革兰染色镜检为蓝紫色阳性球菌。分离菌生化鉴定结果显示，棉籽糖、葡萄糖和甲基红试验呈阳性，触酶试验、硫化氢试验、吲哚试验和V-P试验结果均为阴性，初步鉴定为肠球菌属。16S rDNA测序结果显示，分离株与NCBI登录的海氏肠球菌相似性均在99%以上，确定3株分离株均为海氏肠球菌，分别命名为EH1、EH2和EH3株。EH1、EH2和EH3株对数生长期分别为8～16、8～19和8～14 h，菌液产酸pH均稳定为4.5，具有较好的产酸性能。3株海氏肠球菌在pH为3.0和0.5%胆盐处理下均能存活，具有较好的抗逆性。3株海氏肠球菌无细胞上清液均对大肠杆菌和金黄色葡萄球菌有较好抑菌效果，对链霉素和头孢噻呋呈中介，对环丙沙星、庆大霉素、红霉素、替米考星、青霉素、氨苄西林和氯霉素敏感。以EH1株的自凝集能力较高，为28.00%，以EH2株的疏水性较高，为52.10%；3株海氏肠球菌对昆明小鼠均无致病性。【结论】本研究分离获得3株猎鹰粪源海氏肠球菌，均具有良好的益生性，可作为益生菌制剂的候选菌株。

关键词: 海氏肠球菌; 分离鉴定; 生物学特性; 益生性; 猎鹰

Abstract: 【Objective】 The objective of this study was to isolate lactic acid bacteria with probiotic effect from healthy falcon feces and to provide candidate strains for the development and research of probiotic preparations.【Method】 Six falcon fecal samples were collected from Akqi county,and bacterial isolation and cultivation were carried out using selective culture medium.The isolated bacteria were identified through Gram staining,biochemical identification,16S rDNA PCR amplification,and sequence analysis.Biological characteristics such as growth curve,acid production curve,stress resistance,drug sensitivity,in vitro bacteriostasis,self-aggregation ability,surface hydrophobicity and animal safety test were measured.【Result】 The fecal samples from the falcon yielded three isolates,which were identified as blue-purple positive cocci through Gram staining microscopy.The biochemical analysis demonstrated positive results for raffinose,glucose and methyl red,while the touch enzyme test,hydrogen sulfide test,indole test and V-P test all exhibited negative outcomes.16S rDNA sequencing revealed that the isolates shared over 99% similarity with Enterococcus hirae identified by NCBI.Consequently,the three isolates were classified as Enterococcus hirae and designated EH1,EH2 and EH3,respectively.EH1,EH2 and EH3 strains had logarithmic growth periods of 8-16,8-19 and 8-14 h,respectively.The acid production pH of the bacterial solution was stable at 4.5,indicating good acid production performance.Three strains of Enterococcus hirae were able to survive under pH 3.0 and 0.5% bile salt treatment,demonstrating good stress resistance.The cell-free supernatants of three strains of Enterococcus hirae all showed good antibacterial effects on Escherichia coli and Staphylococcus aureus.They were all mediated by streptomycin and cefotaxifur,and were susceptible to ciprofloxacin,gentamicin,erythromycin,tilmicosin,penicillin,ampicillin and chloramphenicol.The self-agglutination capacity of EH1 strain was higher (28.00%),and the hydrophobic capacity of EH2 strain was higher (52.10%).All of three strains of Enterococcus hirae were non-pathogenic to Kunming mice.【Conclusion】 Three strains of Enterococcus hirae were isolated from falcon feces,which had good probiotics and could be used as candidate strains for probiotic preparations.

Key words: Enterococcus hirae; isolation and identification; biological characteristics; probiotic; falcon

中图分类号:

S831.5

孟竹, 何江, 李静, 李涛, 吴自豪, 陈荣, 齐萌, 冯昕炜. 阿合奇县猎鹰粪源海氏肠球菌的分离鉴定及其生物学特性分析[J]. 中国畜牧兽医, 2024, 51(10): 4587-4595.

MENG Zhu, HEJiang, LIJing, LI Tao, WU Zihao, CHEN Rong, QI Meng, FENG Xinwei. Isolation and Identification of Enterococcus hirae from Falcon Facal in Akqi County and Its Biological Characteristics[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4587-4595.

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