Nayeri Fasaei B, Zahraei Salehi T, Salari S, Ranjbar M M, Yousefi A R
Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
Department of Pathobiology, Faculty of Veterinary Medicine, University of Zabol, Zabol, Iran.
Arch Razi Inst. 2019 Sep;74(3):313-320. doi: 10.22092/ari.2018.120869.1200. Epub 2019 Oct 1.
Colibacillosis is known as a fatal bacterial disease resulting in a high level of commercial loss worldwide. This study amid to elucidate the sequence, genetic characteristics, and phylogeny of the bor gene in Escherichia coli (E. coli) strain c1378 (O78:K80) isolated from avian colibacillosis in Iran and develop a rapid and optimal polymerase chain reaction (PCR) molecular-based technique with specific primers to detect this gene in E. coli. A virulent avian E. coli (i.e., laboratory designation E. coli strain c1378) isolated from a chicken with systemic colibacillosis from a broiler farm in Tehran, Iran, in 2004 was used as a source of the bor gene. After DNA extraction, PCR method was used to amplify the bor gene. A 658 bp fragment of the bor gene was amplified, sequenced, blasted, and phylogenetically studied. The most similar sequences to the bor gene in E. coli strain c1378 were E. coli APEC O78, Enterobacteria phage HK630, and Escherichia coli BW2952, respectively. There was a high similarity between the bor gene in E. coli bacteria with their phage and plasmid. Moreover, a high similarity was observed between the bor and iss genes (approximately 92%) showing that they were homologous genes. In addition, the similarity analysis of different bacterial species, as well as their plasmid and bacteriophage, to the bor gene indicated that the highest similarity to O78:K80 was related to Paracoccidioides brasiliensis, Bacillus thuringiensis CT43 plasmid pBMB0558, and Salmonella enterica subsp. enterica serovar Kentucky strain CVM29188 plasmid, respectively. Altogether, the results of the present study confirmed the presence of the bor gene in the studied isolates and clarified its sequence, phylogenetic relationship, and similarities of E. coli strain c1378 (O78:K80) isolated from avian colibacillosis.
大肠杆菌病是一种致命的细菌性疾病,在全球范围内造成了巨大的商业损失。本研究旨在阐明从伊朗禽大肠杆菌病中分离出的大肠杆菌(E. coli)菌株c1378(O78:K80)中bor基因的序列、遗传特征和系统发育,并开发一种基于聚合酶链反应(PCR)的快速且优化的分子技术,使用特异性引物检测大肠杆菌中的该基因。2004年从伊朗德黑兰一个肉鸡场患有全身性大肠杆菌病的鸡中分离出的一株强毒禽大肠杆菌(即实验室编号为大肠杆菌菌株c1378)被用作bor基因的来源。DNA提取后,采用PCR方法扩增bor基因。扩增出一段658 bp的bor基因片段,进行测序、比对,并进行系统发育研究。大肠杆菌菌株c1378中与bor基因最相似的序列分别是大肠杆菌APEC O78、肠道杆菌噬菌体HK630和大肠杆菌BW2952。大肠杆菌中的bor基因与其噬菌体和质粒之间存在高度相似性。此外,观察到bor基因和iss基因之间有高度相似性(约92%),表明它们是同源基因。此外,对不同细菌物种及其质粒和噬菌体与bor基因的相似性分析表明,与O78:K80相似度最高的分别是巴西副球孢子菌、苏云金芽孢杆菌CT43质粒pBMB0558和肠炎沙门氏菌亚种肠炎血清型肯塔基菌株CVM29188质粒。总之,本研究结果证实了在所研究的分离株中存在bor基因,并阐明了从禽大肠杆菌病中分离出大肠杆菌菌株c1378(O78:K80)的序列、系统发育关系和相似性。