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鸭甲型肝炎病毒1型分离株在实验感染的北京鸭和番鸭雏鸭中的致病性比较

Comparative Pathogenicity of Duck Hepatitis A Virus-1 Isolates in Experimentally Infected Pekin and Muscovy Ducklings.

作者信息

Hisham Islam, Ellakany Hany F, Selim Abdullah A, Abdalla Mohammed A M, Zain El-Abideen Mohamed A, Kilany Walid H, Ali Ahmed, Elbestawy Ahmed R

机构信息

Reference Laboratory for Veterinary Quality Control on Poultry Production (RLQP), Animal Health Research Institute, Damanhour, Egypt.

Poultry and Fish Diseases Department, Faculty of Veterinary Medicine, Damanhour University, Damanhour, Egypt.

出版信息

Front Vet Sci. 2020 May 15;7:234. doi: 10.3389/fvets.2020.00234. eCollection 2020.

Abstract

Duck hepatitis virus (DHV) has always been considered one of the threats endangering duck farming in Egypt since the 1960s. In the current study, suspected DHV samples ( = 30) were obtained from commercial Pekin, Mulard (hybrid), and Muscovy duck farms and backyards in Beheira, Alexandria, Gharbia, Kafr El-Sheikh, and Giza provinces between 2012 and 2017. Diseased 3-21-day-old ducklings showed a clinical history of high mortality rates and nervous signs. Samples were screened by RT-PCR targeting the 5'UTR region and VP1 gene. The PCR-confirmed samples ( = 7) were isolated allantoic route inoculation onto 9-day-old specific-pathogen-free embryonated chicken eggs. Embryos showed stunting, subcutaneous hemorrhages, and liver necrotic greenish-yellow foci. Duck hepatitis A virus-1 (DHAV-1) isolates were genetically analyzed in comparison to other field and vaccine strains. Phylogenetic analyses of the full-length VP1 gene sequences revealed that the obtained DHAV-1 field isolates clustered into genetic group 4 alongside other Egyptian strains isolated during the same period (95.9-99.72% similarity). Amino acid substitutions in the carboxyl-terminal of VP1 (I180T, G184E, D193N, and M213I) were identified in two strains. Also, deletion mutation at I189 was detected in three DHAV-1 strains. Additionally, the two amino acid residues E205 and N235 were common among the isolated strains and other virulent DHAV-1 strains. Two DHAV-1 isolates originated from Pekin source were selected for conducting the comparative pathogenicity testing based on detected point mutations at C-terminus of VP1. We evaluated the pathogenicity of these isolates by investigating clinical signs, mortality rates, and gross pathological and microscopic lesions. The study revealed that experimentally infected Pekin and Muscovy ducklings showed similar clinical signs including squatting down, lateral recumbency, and spasmodic kicking. Muscovy showed milder pathological changes in the liver compared to Pekin ducklings. Histopathological findings supported the gross pathological lesions detected in both breeds. In conclusion, these data provide updated information on the genetic diversity and pathotyping of Egyptian DHAV-1 strains. To the best of our knowledge, this is the first report of comparative pathogenicity of recent DHAV-1 strains in Pekin and Muscovy ducklings in Egypt and the Middle East region.

摘要

自20世纪60年代以来,鸭肝炎病毒(DHV)一直被认为是危及埃及养鸭业的威胁之一。在本研究中,2012年至2017年期间,从贝赫拉、亚历山大、加比亚、卡夫尔谢赫和吉萨省的商业北京鸭、骡鸭(杂交)和番鸭养殖场及后院采集了30份疑似DHV样本。患病的3至21日龄雏鸭表现出高死亡率和神经症状的临床病史。通过针对5'UTR区域和VP1基因的RT-PCR对样本进行筛查。将PCR确认的7份样本通过尿囊途径接种到9日龄无特定病原体的鸡胚中进行分离。胚胎表现出发育迟缓、皮下出血和肝脏坏死的黄绿色病灶。与其他田间和疫苗株相比,对鸭甲型肝炎病毒1型(DHAV-1)分离株进行了基因分析。全长VP1基因序列的系统发育分析表明,获得的DHAV-1田间分离株与同期分离的其他埃及菌株聚集在遗传组4中(相似性为95.9-99.72%)。在两个菌株中鉴定出VP1羧基末端的氨基酸替换(I180T、G184E、D193N和M213I)。此外,在三个DHAV-1菌株中检测到I189处的缺失突变。另外,分离菌株和其他强毒DHAV-1菌株中存在两个氨基酸残基E205和N235。根据在VP1 C末端检测到的点突变,选择了两个源自北京鸭的DHAV-1分离株进行比较致病性试验。我们通过调查临床症状、死亡率以及大体病理和微观病变来评估这些分离株的致病性。研究表明,实验感染的北京鸭和番鸭雏鸭表现出相似的临床症状,包括蹲下、侧卧和痉挛性踢腿。与北京鸭雏鸭相比,番鸭肝脏的病理变化较轻。组织病理学结果支持了在两个品种中检测到的大体病理病变。总之,这些数据提供了关于埃及DHAV-1菌株遗传多样性和致病型的最新信息。据我们所知,这是埃及和中东地区近期DHAV-1菌株在北京鸭和番鸭雏鸭中比较致病性的首次报告。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7607/7326108/974bbdcb8e17/fvets-07-00234-g0001.jpg

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