Al-Hasan Baraa Akeel, Alhatami Abdullah O, Abdulwahab Husam Muhsen, Bustani Ghadeer Sabah, Wahab Alkuwaity Eman Abdul
Medical Laboratory Technology Department, College of Medical Technology, The Islamic University, Najaf, Iraq.
Department of Microbiology, Faculty of Veterinary Medicine, University of Kufa, Najaf, Iraq.
Vet World. 2021 Sep;14(9):2346-2355. doi: 10.14202/vetworld.2021.2346-2355. Epub 2021 Sep 7.
The swollen head syndrome (SHS) makes up complex diseases that infect the upper respiratory tract in poultry and causes several economic losses. Furthermore, this syndrome is considered one of the multifactorial etiological agents. Therefore, this study isolated and molecularly detected (ORT) in poultry.
This study was conducted at 67 broiler farms that had birds observed to be infected with the SHS from September 2018 until August 2019. Subsequently, swabs were collected from their trachea, infraorbital sinuses, and lungs, after which obtained samples were treated through two methods: (a) The direct method, by uploading samples on FTA cards, and the indirect method using a transport media. Afterward, reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the directly treated samples; howeverAQ1, the culture method, followed by PCR, was used to analyze the indirectly treated samples. Next, a partial gene was isolated using four positive PCR products, after which the effect of 16 antibiotics was studied on the seven local ORT strains isolated.
The quantity of ORT isolated using the direct method was 28 (41.7%) samples, which were all positive for the strain. Identification was by direct molecular identification (RT-PCR) from samples loaded on FTA cards. Alternatively, 7 (10.4%) ORTs were detected from the indirect method, as obtained using the culture method and biochemical tests. Then, PCR was subsequently used to confirm the results. As observed, 784 bp bands were shown for all seven ORT isolates. Furthermore, results revealed a significant difference in the detection of ORT strains between direct and indirect methods, with p-value (<0.05) and standard deviation of the error±0.038 for the direct, then ±0.061 for the indirect method. For further analysis on the strain types, four 784 bp PCR products were taken, then partial 16S ribosomal sequence typing was conducted. All these four strains were found to be recorded in NCBI for the 1 time as a local Iraqi strain, with accession numbers (MN931657, MN931656, MN931655, and MN931654). Notably, results also showed that all isolated strains were multidrug-resistant.
From the results, ORT is proposed to be implicated as one of the etiological factors that cause SHSs in poultry. Phylogenetic analysis of the current ORT bacterial strains also showed that they are closely related to the Egyptian isolates.
肿头综合征(SHS)是一种感染家禽上呼吸道的复杂疾病,会造成多种经济损失。此外,该综合征被认为是多因素致病因子之一。因此,本研究对家禽中的禽呼肠孤病毒(ORT)进行了分离和分子检测。
本研究在67个肉鸡养殖场开展,这些养殖场的鸡在2018年9月至2019年8月期间被观察到感染了SHS。随后,从鸡的气管、眶下窦和肺中采集拭子,采集的样本通过两种方法进行处理:(a)直接法,将样本加载到FTA卡上;间接法,使用运输培养基。之后,采用逆转录聚合酶链反应(RT-PCR)分析直接处理的样本;然而,对于间接处理的样本,先采用培养法,然后进行PCR分析。接下来,使用四个阳性PCR产物分离出部分基因,之后研究了16种抗生素对分离出的7株本地ORT菌株的作用。
采用直接法分离出ORT的样本数量为28个(41.7%),所有菌株均为阳性。通过对加载在FTA卡上的样本进行直接分子鉴定(RT-PCR)来进行鉴定。另外,采用培养法和生化试验从间接法中检测到7株(10.4%)ORT。随后使用PCR对结果进行确认。如观察到的,所有7株ORT分离株均显示出784 bp的条带。此外,结果显示直接法和间接法在ORT菌株检测上存在显著差异,直接法的p值(<0.05)和误差标准偏差为±0.038,间接法为±0.061。为了进一步分析菌株类型,选取了四个784 bp的PCR产物,然后进行部分16S核糖体序列分型。发现所有这四株菌株在NCBI上首次被记录为伊拉克本地菌株,登录号为(MN931657、MN931656、MN931655和MN931654)。值得注意的是,结果还表明所有分离菌株均具有多重耐药性。
根据结果,ORT被认为是导致家禽SHS的病因之一。对当前ORT细菌菌株的系统发育分析还表明,它们与埃及分离株密切相关。