肉牛呼吸道皮特不动杆菌的生物学特性分析

doi:10.3969/j.issn.1004-1524.2022.02.11

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (2): 300-309.DOI: 10.3969/j.issn.1004-1524.2022.02.11

肉牛呼吸道皮特不动杆菌的生物学特性分析

樊利虹¹(), 王之盛², 左之才¹^,^*(), 才冬杰¹, 易军³, 马晓平¹, 苟丽萍¹, 王巍³

1.四川农业大学动物医学院,四川成都 611130
2.四川农业大学动物营养研究所,四川成都 611130
3.四川省畜牧科学研究院,四川成都 610066

收稿日期:2020-10-21 出版日期:2022-02-25 发布日期:2022-03-02
通讯作者: 左之才
作者简介:左之才,E-mail: zzcjl@126.com
樊利虹(1996—),女,四川成都人,硕士研究生,研究方向为中西兽医与临床。E-mail: 2323702149@qq.com
基金资助:
国家现代农业产业技术体系(肉牛牦牛,CARS-37);国家现代农业产业技术体系四川肉牛创新团队项目(SCCXTD-2020-13)

Analysis of biological characteristics of Acinetobacter pittii from respiratory tract of beef cattle

FAN Lihong¹(), WANG Zhisheng², ZUO Zhicai¹^,^*(), CAI Dongjie¹, YI Jun³, MA Xiaoping¹, GOU Liping¹, WANG Wei³

1. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China
3. Sichuan Academy of Animal Husbandry,Chengdu 610066,China

Received:2020-10-21 Online:2022-02-25 Published:2022-03-02
Contact: ZUO Zhicai

摘要/Abstract

摘要：

为探究分离自四川南充、宜宾和什邡3个地区肉牛呼吸道的皮特不动杆菌之间生物学特性的差异,本实验对5株皮特不动杆菌分别进行了细菌染色、培养特性、生化试验、16S rDNA和gyrB基因序列聚类分析以及MLST分型等研究。结果显示,5株分离自肉牛呼吸道皮特不动杆菌的菌体形态和不同培养基上的菌落形态均相似;5株菌的生化特性并非完全一致,主要表现在ZZCSF1807-9的枸橼酸盐、精氨酸双水解试验为阳性,丙二酸盐为阴性,与其余4株菌相反;16S rDNA系统进化树显示,5株菌均聚集在同一分支,而gyrB基因系统进化树显示,ZZCSF1807-9与其余4株菌未在同一分支,存在进化距离;MLST分型结果显示,ZZCSF1807-9为ST214型,而另4株菌均与ST321型最为接近。上述结果表明,来源于呼吸道疾病肉牛的5株皮特不动杆菌在生化特性表型和gyrB基因,以及MLST型上存在一定差异,发生这些差异是否与各菌株的环境适应或因毒力变化有关尚有待进一步研究。

关键词: 皮特不动杆菌, 生化特性, 同源性分析, 多位点序列分型(MLST)

Abstract:

In order to explore the differences in biological characteristics of Acinetobacter pittii isolated from respiratory tract of beef cattle in Nanchong, Yibin, and Shifang in Sichuan Province, five strains of Acinetobacter pittii were studied by bacterial staining, culture characteristics, biochemical test, 16S rDNA and gyrB gene sequence clustering analysis and MLST typing. The results showed that the mycelial morphology and the colonial morphology of 5 strains of Acinetobacter pittii on different media were similar. And the biochemical characteristics of the five strains were not completely consistent, which was mainly manifested in the citrate and arginine double hydrolysis test of ZZCSF1807-9, while malonate was negative, contrary to the other four strains. The 16S rDNA phylogenetic tree showed that all 5 strains were clustered in the same branch, while the gyrB phylogenetic tree showed that ZZCSF1807-9 was not in the same branch with the other 4 strains, and there was an evolutionary distance. MLST typing results showed that ZZCSF1807-9 was ST214, while the other 4 strains were the closest to ST321.The above results indicated that the five strains of Acinetobacter pittii derived from beef cattle with respiratory disease had certain differences in biochemical characteristic phenotype, gyrB gene and MLST type, and whether the differences were related to the environmental adaptation of each strain or the change in virulence remained to be further studied.

Key words: Acinetobacter pittii, biochemical characteristics, homology analysis, multilocus sequence typing (MLST)

中图分类号:

S858.23

樊利虹, 王之盛, 左之才, 才冬杰, 易军, 马晓平, 苟丽萍, 王巍. 肉牛呼吸道皮特不动杆菌的生物学特性分析[J]. 浙江农业学报, 2022, 34(2): 300-309.

FAN Lihong, WANG Zhisheng, ZUO Zhicai, CAI Dongjie, YI Jun, MA Xiaoping, GOU Liping, WANG Wei. Analysis of biological characteristics of Acinetobacter pittii from respiratory tract of beef cattle[J]. Acta Agriculturae Zhejiangensis, 2022, 34(2): 300-309.

图/表 13

参考文献 27

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基因名称 Gene name	引物序列 Primer sequence (5'-3')	片段长度 Product length/bp
16S rDNA	P1:AGAGTTTGATCCTGGCTCAG P2:TACGGCTACCTTGTTACGACTT	1500
gyrB	P3:ACYTTCTTCTTCCGTCAAATG P4:CCTTTATARCGYTGRATCATC	701

基因名称 Gene name	引物序列 Primer sequence (5'-3')	片段长度 Product length/bp
16S rDNA	P1:AGAGTTTGATCCTGGCTCAG P2:TACGGCTACCTTGTTACGACTT	1500
gyrB	P3:ACYTTCTTCTTCCGTCAAATG P4:CCTTTATARCGYTGRATCATC	701

生化项目Biochemical index	ZZCNC1807-4	ZZCNC1807-6	ZZCNC1807-7	ZZCJL1807-8	ZZCSF1807-9
蔗糖Sucrose	-	+	+	-	-
乳糖Lactose	-	-	+	-	-
甘露醇Mannitol	-	+	+	-	-
枸橼酸盐Citrate	-	-	-	-	+
丙二酸盐Lonate	+	+	+	+	-
精氨酸双水解酶 Arginine dihydrolase	-	-	-	-	+
MR	-	+	-	-	-

生化项目Biochemical index	ZZCNC1807-4	ZZCNC1807-6	ZZCNC1807-7	ZZCJL1807-8	ZZCSF1807-9
蔗糖Sucrose	-	+	+	-	-
乳糖Lactose	-	-	+	-	-
甘露醇Mannitol	-	+	+	-	-
枸橼酸盐Citrate	-	-	-	-	+
丙二酸盐Lonate	+	+	+	+	-
精氨酸双水解酶 Arginine dihydrolase	-	-	-	-	+
MR	-	+	-	-	-

菌株名称 Strain name	GenBank登录号 Accession No.	来源 Isolation source	分离地 Separated area	时间 Separated time
218	KM281497	血液(人)Blood (Homo sapiens)	伊朗Iran	2014
EC3N45	MK348732	太阳鸟泄殖腔液Sunbird cloacal fluid	以色列Israel	2019
SN6-2	LC485224	豇豆Vigna unguiculatavar. sesquipedalis	泰国Thailand	2019
NQ-001	KY921876	泥鳅Misgurnus anguillicaudatus	中国China	2017
NF108343	JQ040019	痰液Sputum	中国China	2012
15c	LC499764	城市污水Municipal wastewater	印度India	2019
CW-17	MG009235	麦田Crop field	中国China	2017
CS-1	KY828845	根际土壤Rhizospheric soil	巴基斯坦Pakistan	2018
W26	KY922994	武昌鱼Megalobrama amblycephala	中国China	2017
CIACST3	MG786615	鲢鱼Hypophthalmichthys molitrix	印度India	2018
DRLL11	KU550179	白纹伊蚊幼虫Larvae midgut	印度India	2016
BUYA-7	MN490071	石油污染土壤Oil-contaminated soil	蒙古Mongolia	2019

肉牛呼吸道皮特不动杆菌的生物学特性分析

Analysis of biological characteristics of Acinetobacter pittii from respiratory tract of beef cattle

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菌株名称 Strain name	GenBank登录号 Accession No.	分离源 Isolation source	分离地 Separated area	时间 Separated time
AP-882	CP014477	伤口拭子Wound swab	马来西亚Malaysia	2014
NQ-003	CP035109	鱼Fish	中国(河南)Henan, China	2013
C54	CP042364	临床样品Clinical sample	澳大利亚Australia	2014
YMC2010/81T346	CP017938	腹膜导管尖端Peritoneal catheter tip	韩国South Korea	2014
DUT-2	CP014651	海洋沉积物Marine sediments	中国China	2015
IEC338SC	CP015145	气管分泌物(人)Tracheal secretion (Homo sapiens)	巴西Brazil	2014
WCHAP005069	CP026089	临床样品(人)Clinical sample (Homo sapiens)	中国(成都)Chengdu, China	2018
AP43	CP043052	尿液(人)Urine	中国(杭州)Hangzhou, China	2018
XJ88	CP018909	痰液(人)Sputum (Homo sapiens)	中国(成都)Chengdu, China	2013
ST220	CP029610	痰液(人)Sputum (Homo sapiens)	中国(合肥)Hefei, China	2017

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