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苏丹东部小反刍动物中流行的口疮病毒(ORFV)和单峰骆驼中伪牛痘病毒(PCPV)的现场调查及系统发育特征分析

Field investigation and phylogenetic characterization of orf virus (ORFV) circulating in small ruminants and Pseudocowpoxvirus (PCPV) in dromedary camels of eastern Sudan.

作者信息

Khalafalla Abdelmalik Ibrahim, Elhag Ahmed Eisa, Ishag Hassan Zackaria Ali

机构信息

Department of Microbiology, Faculty of Veterinary Medicine, University of Khartoum, P. O. Box 32, Khartoum North, Shambat, Sudan.

Veterinary Laboratories Division, Animal Wealth Sector, Abu Dhabi Agriculture and Food Safety Authority P. O. Box 52150, Abu Dhabi, United Arab Emirates.

出版信息

Heliyon. 2020 Mar 21;6(3):e03595. doi: 10.1016/j.heliyon.2020.e03595. eCollection 2020 Mar.

Abstract

In this study, livestock herders in eastern Sudan were interviewed through structured questionnaire involved 14046 animals in 151 herds (87 camel herds, 51 sheep and 13 goats) from June to September of 2016 in Showak area of Gadarif State to get some epidemiological information on contagious ecthyma (CE) infection. 102 suspected cases of CE were investigated (38 sheep, 22 goats and 42 camels) by a second questionnaire focusing on age and sex of affected animals beside number and localization of the lesions. Representative tissue samples of scab lesion scrapings were collected from a total of 36 suspected sheep, goats and camels for DNA extraction to identify PPV by quantitative real-time PCR and gel-based PCR, then a PCR protocol was used to obtain DNA fragment of B2L gene from six DNAs (2 from each animal species) for sequencing. Phylogenetic tree based on nucleotide sequences was constructed and all data were analyzed statistically. Obtained result has shown morbidity rate of 23.8% and a case fatality rate of 4.7 % in overall investigated animals resulting in a significant economic loss. Within individual herd, the morbidity rate varied from 5.6 to 42.8%, while the case fatality rate ranged between 0 and 33.3%. Camels accounted for the highest case fatality rate with 6.5% compared to sheep and goats which their rates were 2.8% and 1.3%, respectively. 93% of the affected animals were young less than one-year-old. The prevalence of CE was high in the rainy season compared to winter and summer. Out of 36 scab materials collected from sheep, goats, and camels, 24 gave positive specific amplification in real-time PCR and 21 in the gel-based PCR. DNA sequencing confirmed the PCR results. All sequences had a high G + C content of 62.6-63.9%. A BLAST search also revealed that the studied sheep PPV (SPPV) isolates shared 99.08% nucleotide sequence intragroup identity, 96.88-97.27% identity with the goat PPV (GPPV) isolates and together they belong to the Orf virus (ORFV) species, while the camel PPV (CPPV) isolates are close to the Pseudocowpoxvirus (PCPV) species of the PPV genus and share 92.51-93.62 % identity with the GPPV isolates. In conclusion the present study demonstrated that the gross lesion produced by PPV in sheep, goats and camels is generally similar, yet the PPVs circulating in eastern Sudan in camels (PCPV) are genetically distinct from those affecting sheep and goats (ORFV). Contagious ecthyma in eastern Sudan causes significant morbidities and mortalities and control measures, guided by the results of this investigation ought to be implemented.

摘要

在本研究中,2016年6月至9月期间,在加达里夫州绍瓦克地区通过结构化问卷对苏丹东部的家畜牧民进行了访谈,这些问卷涉及151个畜群中的14046只动物(87个骆驼群、51个绵羊群和13个山羊群),以获取有关传染性脓疱病(CE)感染的一些流行病学信息。通过第二份问卷对102例CE疑似病例(38只绵羊、22只山羊和42头骆驼)进行了调查,该问卷除了关注病变的数量和位置外,还侧重于受影响动物的年龄和性别。从总共36只疑似绵羊、山羊和骆驼中采集了痂皮病变刮屑的代表性组织样本用于DNA提取,通过定量实时PCR和基于凝胶的PCR鉴定副痘病毒(PPV),然后使用PCR方案从六个DNA(每种动物2个)中获得B2L基因的DNA片段进行测序。构建了基于核苷酸序列的系统发育树,并对所有数据进行了统计分析。获得的结果显示,在所有调查动物中,发病率为23.8%,病死率为4.7%,造成了重大经济损失。在单个畜群中,发病率从5.6%到42.8%不等,而病死率在0到33.3%之间。骆驼的病死率最高,为6.5%,而绵羊和山羊的病死率分别为2.8%和1.3%。93%的受影响动物为不到一岁的幼畜。与冬季和夏季相比,CE在雨季的患病率较高。从绵羊、山羊和骆驼采集的36份痂皮材料中,24份在实时PCR中呈阳性特异性扩增,21份在基于凝胶的PCR中呈阳性。DNA测序证实了PCR结果。所有序列的G + C含量都很高,为62.6 - 63.9%。BLAST搜索还显示,所研究的绵羊PPV(SPPV)分离株在组内核苷酸序列一致性为99.08%,与山羊PPV(GPPV)分离株的一致性为96.88 - 97.27%,它们共同属于口疮病毒(ORFV)种,而骆驼PPV(CPPV)分离株与PPV属的假牛痘病毒(PCPV)种接近,与GPPV分离株的一致性为92.51 - 93.62%。总之,本研究表明,PPV在绵羊、山羊和骆驼中产生的肉眼病变通常相似,但在苏丹东部骆驼中传播的PPV(PCPV)在基因上与影响绵羊和山羊的PPV(ORFV)不同。苏丹东部的传染性脓疱病导致了重大的发病和死亡,应根据本调查结果实施控制措施。

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