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埃塞俄米亚奥罗米亚中部与犊牛相关的冠状病毒和轮状病毒的分离与鉴定

Isolation and Characterization of Coronavirus and Rotavirus Associated with Calves in Central Part of Oromia, Ethiopia.

作者信息

Seid Umer, Dawo Fufa, Tesfaye Asamino, Ahmednur Munera

机构信息

College of Agriculture, Oda Bultum University, P.O. Box 226, Chiro, Ethiopia.

College of Veterinary Medicine, Addis Ababa University, P.O. Box 34, Bishoftu, Ethiopia.

出版信息

Vet Med Int. 2020 Nov 30;2020:8869970. doi: 10.1155/2020/8869970. eCollection 2020.

Abstract

BACKGROUND

Coronavirus and rotavirus are most commonly associated etiologies for calves' diarrhoea, resulting in loss of productivity and economy of farmers. However, various facets of diarrheal disease caused by coronavirus and rotavirus in calves in Ethiopia are inadequately understood. A cross-sectional study was conducted with the aim of isolation and molecular characterization of coronavirus and rotavirus from calves in the central part of Oromia (Bishoftu, Sebata, Holeta, and Addis Ababa), Ethiopia, from November 2018 to May 2019. The four study areas were purposively selected and faecal samples were collected by simple random sampling for diagnosis of coronavirus and rotavirus infection by using the antigen detection enzyme-linked immunosorbent assay (Ag-ELISA) kit. In addition, this study was carried out to have insight in prevalence and associated risk factors of coronavirus and rotavirus infection in calves.

RESULT

During the study, 83 diarrheic and 162 nondiarrheic faecal samples collected from calves less than 4 weeks of age were screened for coronavirus and rotavirus. Of the 83 diarrheic samples, 1 sample (1.2%) was positive for coronavirus antigen and 6 samples (7.2%) were found to be positive for rotavirus antigen by Ag-ELISA. All the nondiarrheic samples were negative for both coronavirus and rotavirus Ag. The overall prevalence of coronavirus and rotavirus infection in calves was estimated at 0.4% (1/245) and 2.45% (6/245), respectively. All samples (7) of ELISA test positive of both coronavirus and rotavirus were propagated in Madin-Darby bovine kidney (MDBK) cells. After 3 subsequent passages, progressive cytopathic effect (CPE), i.e., rounding, detachment, and the destruction of monolayer cell of five samples (1 sample of coronavirus and 4 samples of rotavirus) (71.4%) were observed. At the molecular stage, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to determine the presence of coronavirus and rotavirus nucleic acid by using specific primers. The 5 samples that were coronavirus and rotavirus antigen positive by ELISA and develop CPE on cell culture were also positive on RT-PCR technique. The prevalence of infection peaked at 1st and 2nd weeks of age in male calves.

CONCLUSION

Diarrheal disease caused by coronavirus and rotavirus has a great health problem in calves that interrupts production benefits with reduced weight gain and increased mortality and its potential for zoonotic spread. So, the present findings show coronavirus and rotavirus infection in calves in Ethiopia that needs to be addressed by practising early colostrum feeding in newborn calves, using vaccine, or improving livestock management.

摘要

背景

冠状病毒和轮状病毒是犊牛腹泻最常见的相关病因,会导致农民生产力下降和经济损失。然而,埃塞俄比亚犊牛由冠状病毒和轮状病毒引起的腹泻疾病的各个方面尚未得到充分了解。2018年11月至2019年5月,在埃塞俄比亚奥罗米亚中部(比绍夫图、塞巴塔、霍莱塔和亚的斯亚贝巴)开展了一项横断面研究,旨在从犊牛中分离冠状病毒和轮状病毒并进行分子特征分析。这四个研究区域是有目的地选择的,通过简单随机抽样收集粪便样本,使用抗原检测酶联免疫吸附测定(Ag-ELISA)试剂盒诊断冠状病毒和轮状病毒感染。此外,开展本研究是为了深入了解犊牛冠状病毒和轮状病毒感染的患病率及相关风险因素。

结果

在研究期间,对从4周龄以下犊牛收集的83份腹泻粪便样本和162份非腹泻粪便样本进行了冠状病毒和轮状病毒筛查。在83份腹泻样本中,通过Ag-ELISA检测,1份样本(1.2%)冠状病毒抗原呈阳性,6份样本(7.2%)轮状病毒抗原呈阳性。所有非腹泻样本的冠状病毒和轮状病毒抗原均为阴性。犊牛冠状病毒和轮状病毒感染的总体患病率分别估计为0.4%(1/245)和2.45%(6/245)。所有冠状病毒和轮状病毒ELISA检测均为阳性的7份样本在麦迪逊-达比牛肾(MDBK)细胞中进行传代培养。经过3次连续传代后,观察到5份样本(1份冠状病毒样本和4份轮状病毒样本)(71.4%)出现进行性细胞病变效应(CPE),即细胞变圆、脱落和单层细胞破坏。在分子阶段,使用逆转录聚合酶链反应(RT-PCR)技术,通过特异性引物确定冠状病毒和轮状病毒核酸的存在。ELISA检测冠状病毒和轮状病毒抗原呈阳性且在细胞培养中出现CPE的5份样本RT-PCR检测也呈阳性。雄性犊牛感染率在1周龄和2周龄时达到峰值。

结论

冠状病毒和轮状病毒引起的腹泻疾病对犊牛健康构成重大问题,但会因体重增加减少、死亡率增加及其人畜共患病传播潜力而影响生产效益。因此,目前的研究结果表明埃塞俄比亚犊牛存在冠状病毒和轮状病毒感染问题,需要通过对新生犊牛尽早饲喂初乳、使用疫苗或改善畜牧管理来解决。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4e43/7723472/df9bc169f9d8/VMI2020-8869970.001.jpg

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