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孟加拉国奶牛中布鲁氏菌流产亚种生物变种3的首次分离、鉴定及基因特征分析

First isolation, identification and genetic characterization of Brucella abortus biovar 3 from dairy cattle in Bangladesh.

作者信息

Islam Md Sadequl, Garofolo Giuliano, Sacchini Lorena, Dainty Amanda C, Khatun Mst Minara, Saha Sukumar, Islam Md Ariful

机构信息

Department of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh, Bangladesh.

Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

出版信息

Vet Med Sci. 2019 Nov;5(4):556-562. doi: 10.1002/vms3.193. Epub 2019 Aug 26.

Abstract

BACKGROUND

Brucellosis is a zoonotic disease caused by bacteria Brucella spp. belonging to the genus Brucella. It is endemic in domesticated animals in Bangladesh. Isolation, identification and genetic characterization of Brucella spp. in dairy cattle are essential to undertake appropriate control and preventive measures. The study was conducted to isolate and characterize the Brucella spp. circulating in dairy cattle.

METHODS

Uterine discharge (n = 45), milk (n = 115), vaginal swab (n = 71), placenta (n = 7) and aborted fetus (n = 2) were collected. Brucella selective agar plates were inoculated with samples and incubated at 37 C for 14 days under 5% CO for isolation of Brucella spp. Brucella suspected colonies were recovered from samples were confirmed by genus and species specific PCR assays. Genetic characterization was performed by Multi Locus Variable number tandem-repeat Analysis-16 (MLVA-16).

RESULTS

The isolates of Brucella recovered from samples were confirmed as B. abortus by AMOS-ERY PCR assay. The classical biotyping method confirmed all 10 B. abortus isolates belonged to the biovar 3. The MLVA-16 assay indicated all B. abortus isolates identical and the same genotype 40, based on panel 1 MLVA-8.

CONCLUSION

Dendrogram analysis revealed all B. abortus isolates of the study were identical to three isolates from Brazil, one isolate of France and closely related to Chinese isolates. This is the first report of isolation and genetic characterization of B. abortus from the dairy cattle in Bangladesh.

摘要

背景

布鲁氏菌病是一种由布鲁氏菌属细菌引起的人畜共患病。在孟加拉国,它在家畜中呈地方性流行。对奶牛中的布鲁氏菌进行分离、鉴定和基因特征分析对于采取适当的控制和预防措施至关重要。本研究旨在分离和鉴定在奶牛中传播的布鲁氏菌。

方法

采集子宫分泌物(n = 45)、牛奶(n = 115)、阴道拭子(n = 71)、胎盘(n = 7)和流产胎儿(n = 2)。将样品接种到布鲁氏菌选择性琼脂平板上,在37℃、5%二氧化碳条件下培养14天以分离布鲁氏菌。从样品中回收的疑似布鲁氏菌菌落通过属和种特异性PCR检测进行确认。通过多位点可变数目串联重复分析-16(MLVA-16)进行基因特征分析。

结果

通过AMOS-ERY PCR检测,从样品中分离出的布鲁氏菌被确认为流产布鲁氏菌。经典生物分型方法证实所有10株流产布鲁氏菌均属于生物变种3。基于MLVA-8的第1组,MLVA-16检测表明所有流产布鲁氏菌分离株相同且具有相同的基因型40。

结论

聚类分析表明,本研究中的所有流产布鲁氏菌分离株与来自巴西的三株分离株、法国的一株分离株相同,并且与中国分离株密切相关。这是孟加拉国首次关于从奶牛中分离出流产布鲁氏菌并进行基因特征分析的报告。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9541/6868452/dc9723854f20/VMS3-5-556-g001.jpg

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