Molecular Identification and Phylogenetic Analysis of Salmonella‎ species ‎Isolated from Diarrheal Children and Dogs in Baghdad Governorate, ‎Iraq‎

Fudhaa Abdhalla, Nagham M Al-Gburi
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Abstract

This work aimed to use conventional PCR to identify Salmonella‎ spp. that ‎were isolated from diarrheal children and healthy and diarrheic dogs based on four ‎virulence genes, hilA, stn, spvR‎, and marT. Sixteen Salmonella‎ isolates including: 9 ‎isolated from children's diarrhea from three species (S. Typhimurium, S. Enteritidis, S. ‎Typhi) and seven isolated from dogs including (S. Typhimurium, S. Enteritidis, S. ‎Muenchen), were identified primarily by several methods. The PCR products of the 16S ‎rRNA gene were sequenced and examined using BLAST analysis to find differences and ‎similarities between these Iraqi isolates and already-known global strains in order to ‎construct the phylogenetic tree of S. Muenchen which was detected for the first time in ‎dogs in Iraq. The results of the study revealed that all isolates of Salmonella‎ obtained ‎from children possess the hilA and stn genes. The marT gene was detected in 88% of the ‎Salmonella‎ serovars, and the spvR‎ gene was carried in 55% of the isolates. Among dog ‎Salmonella‎ isolates, the hilA gene was detected at 100%, the stn gene was at 85.7%, the ‎marT gene was present at 71.4%, while the spvR‎ gene was found at 57.1%. The result of ‎DNA sequencing and phylogenetic tree indicated that the local Iraqi S. Muenchen was ‎extremely close to the national strain and share the same ‎16S rRNA gene sequence, the ‎isolate was registered at NCBI and became a global reference with the accession number OQ999043.1. In conclusion, the presence of these important virulence genes among ‎Salmonella‎ serovars isolated from children and dogs alerted on the potential risk of ‎contamination of the environment and may lead to a community health crisis‎.
从伊拉克巴格达省腹泻儿童和狗身上分离出的沙门氏菌的分子鉴定和系统发育分析
这项工作旨在利用传统的 PCR 技术,根据 hilA、stn、spvR 和 marT 四个毒力基因来鉴定从腹泻儿童和健康及腹泻犬中分离出来的沙门氏菌属。16 个沙门氏菌分离物包括9 株分离自儿童腹泻的三种沙门氏菌(鼠伤寒沙门氏菌、肠炎沙门氏菌、伤寒沙门氏菌)和 7 株分离自狗的沙门氏菌(鼠伤寒沙门氏菌、肠炎沙门氏菌、门冬沙门氏菌),主要通过几种方法进行鉴定。对 16S rRNA 基因的 PCR 产物进行了测序,并使用 BLAST 分析法对这些伊拉克分离物与全球已知菌株之间的异同进行了检验,从而构建了首次在伊拉克狗体内检测到的 S. Muenchen 的系统发生树。研究结果表明,从儿童身上分离出的所有沙门氏菌都具有 hilA 和 stn 基因。88% 的沙门氏菌血清型中检测到 marT 基因,55% 的分离株携带 spvR 基因。在狗沙门氏菌分离物中,hilA 基因的检出率为 100%,stn 基因的检出率为 85.7%,marT 基因的检出率为 71.4%,而 spvR 基因的检出率为 57.1%。DNA 测序和系统发生树的结果表明,当地的伊拉克 S. Muenchen 与国家菌株极为接近,具有相同的 16S rRNA 基因序列,该分离株已在 NCBI 注册,并成为全球参考株,登录号为 OQ999043.1。总之,从儿童和狗身上分离出的沙门氏菌血清型中存在这些重要的毒力基因,提醒人们注意环境污染的潜在风险,并可能导致社区健康危机。
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