Molecular Detection of Virulence Genes in Escherichia coli and Salmonella enterica Isolated from Minimally Processed Foods Sold within Kaduna Metropolis

A. Haroun, Hafsat A., Kamaluddeen Kabir, D. Denwe
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Abstract

This study was aimed at detecting virulence genes in enteropathogenic bacteria associated with minimally processed foods sold within Kaduna metropolis. Samples of sliced watermelons, peeled sugarcanes, peeled and sliced pineapples and unshelled coconuts (n=140) were collected from Kaduna North, Kaduna South and Igabi LGAs. Upon enrichment, samples were inoculated onto Eosin-Methylene Blue (EMB) and Cefixime-Tellurite-Sorbitol MacConkey (CT-SMAC) agar and incubated for 18 hours at 35oC. Isolates were identified using MicrobactTM 24E Gram Negative Bacteria Identification System for Enterobacteriaceae. Thereafter, E. coli and Salmonella enterica isolates’ DNA was extracted and purified using AccuPrep Genomic DNA Extraction Kit; used as a template for the PCR amplification of the bundle forming pilus (bfpA) and invasion (invA) genes; and then the PCR products were visualised using agarose gel electrophoresis documentation system. The results of the MicrobactTM analyses showed widespread contamination of the samples with E. coli (22 isolates) and Salmonella enterica (3 isolates). There was no statistically significant difference in the bacterial contaminants isolated from the various sampling areas (one-way ANOVA: p = 0.577); similarly, the type of the minimally processed food samples had no influence on the prevalence of E. coli and Salmonella enterica isolates (p = 0.345). PCR result revealed that the invA virulence gene (284bp) was present in one Salmonella enterica isolate. The presence of invA gene in the Salmonella enterica isolate indicated that the isolate is a virulent strain, which can cause food-borne infections. These results indicated the contamination of the minimally processed fruits with enteropathogens, hence, standards of quality control should be enshrined, towards safer foods and enhanced health of the consuming populace. Keywords: E. coli; virulence gene; minimally processed foods; Salmonella enterica.
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卡杜纳市市售微加工食品中大肠杆菌和肠炎沙门氏菌毒力基因的分子检测
本研究旨在检测与卡杜纳大都市内销售的最低限度加工食品相关的肠致病性细菌的毒力基因。从北卡杜纳、南卡杜纳和伊加比地方政府收集了切片西瓜、去皮甘蔗、去皮菠萝和去壳椰子的样本(n=140)。富集后,将样品接种于伊红-亚甲基蓝(EMB)和头孢昔肟- tellite -山梨醇MacConkey (CT-SMAC)琼脂上,在35℃下孵育18小时。采用MicrobactTM 24E革兰氏阴性菌鉴定系统对分离菌株进行鉴定。随后,采用AccuPrep基因组DNA提取试剂盒对大肠杆菌和肠沙门氏菌分离株进行DNA提取和纯化;作为PCR扩增成束菌毛(bfpA)和侵染(invA)基因的模板;然后用琼脂糖凝胶电泳记录系统将PCR产物可视化。MicrobactTM分析结果显示,样品广泛受到大肠杆菌(22株)和肠沙门氏菌(3株)的污染。各采样区分离的细菌污染物无统计学差异(单因素方差分析:p = 0.577);同样,最低限度加工食品样品的类型对大肠杆菌和分离肠沙门氏菌的流行率没有影响(p = 0.345)。PCR结果显示,1株肠沙门氏菌分离株存在284bp的invA毒力基因。肠道沙门氏菌分离物中存在invA基因,表明该分离物是一种可引起食源性感染的毒力菌株。这些结果表明,经过最低限度加工的水果受到肠致病菌的污染,因此,应制定质量控制标准,以实现更安全的食品和提高消费人群的健康。关键词:大肠杆菌;毒力基因;最低限度加工食品;沙门氏菌血清。
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