Molecular Characterisation of Gut Bacteria in Wista Rat After Green Tea Consumption

Joyce Uweh Emmanuel, Agi Vivian Nkemkanma, Aleru Prisca Constancy
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Abstract

Green tea can influence the gut bacteria by either stimulating or hindering the growth of specific species. This experimental study was carried out in Rivers State, Nigeria to molecularly characterize gut bacteria in Wista rats after green tea consumption. Twenty-five (25) rats divided into five groups were used; each group had five (5) rats having an average weight range of 70g – 130g. After acclimatization of 2 weeks, groups (2, 3, 4 & 5) were feed with green tea extract based on standard method using oral gavage with rat feed pellet and water for seven (7) days and fourteen (14) days respectively based on their body weights and group 1 (control) was fed with rat feed pellet and water only for same duration and their weights recorded. Stool samples were collected aseptically by pulling from each group into sterile stool sample bottles and sent to the laboratory for immediate analysis. Conventional microbiological methods of culturing and molecular methods were utilized for identification of isolates, total heterotrophic plate count and antibiotic susceptibility were also done. The data from this study were analysed using SPSS version 23, to survey difference in gut bacterial count of wista rats after fed with green tea, rat weight across the study groups, and the values were expressed as mean ± SD at a 5% alpha level. The study reported a statistical difference in bacterial plate count at (P≤0.05) in Groups 2, 3, 4, and 5 after 14 Days of Acclimatization, after 7 days of Treatment and after further 14 Days of Treatment. Group 1 showed no statistical significance at P=0.212. Furthermore, there was significant difference in weight comparison at (P≤0.05) in rats contained in all groups after 7 days and after further 14 days of treatment, rats in group 1 showed weight gain. There was weight loss in all the treatment groups (2 ,3, 4, and 5). Antibiotic-resistant assay pattern of the isolates showed multiple drugs resistance to all the antibiotics used with Bacillus and Enterobacter being the most resistant isolates. The molecular characterization and identification of the isolates revealed close relatedness to Staphylococcus arlettae, Enterobacter cloacae, Enterobacter cancerogenus, Acinetobacter nosocomialis, Comamonas testosterone, Klebsiella veriicola, Bacillus thuringiensis, Bacillus pumilus and Bacillus cereus, it showed that the molecular technique proved to be more specific and accurate than the convectional technique in the identification and characterization of the isolates. Also, the gut bacterial count of the rats that were treated with the tea were significantly raised indicating that the tea has the potential to influence the abundance and diversity of gut bacteria. As seen in this study, molecular characterisation of gut bacteria identifies bacteria upto their specie level and they are influenced by diet.
饮用绿茶后 Wista 大鼠肠道细菌的分子特征
绿茶可以通过刺激或阻碍特定物种的生长来影响肠道细菌。这项实验研究在尼日利亚河流州进行,目的是对饮用绿茶后 Wista 大鼠肠道细菌的分子特征进行分析。25 只大鼠分为 5 组,每组 5 只,平均体重在 70 克至 130 克之间。适应 2 周后,根据大鼠体重,按照标准方法,用大鼠饲料颗粒和水口服绿茶提取物,分别饲喂 2、3、4 和 5 组七(7)天和十四(14)天。在无菌的情况下,从每组中抽取粪便样本,装入无菌粪便样本瓶中,并立即送往实验室进行分析。利用传统的微生物学培养方法和分子方法对分离物进行鉴定,还进行了异养菌平板总数和抗生素敏感性分析。本研究的数据使用 SPSS 23 版进行分析,以调查 Wista 大鼠在喂食绿茶后肠道细菌数量的差异,以及各研究组大鼠体重的差异,数值以平均值 ± SD 表示,α水平为 5%。研究报告显示,在适应 14 天后、治疗 7 天后和继续治疗 14 天后,第 2、3、4 和 5 组的细菌平板计数在统计学上存在差异(P≤0.05)。第 1 组没有统计学意义,P=0.212。此外,各组大鼠在治疗 7 天后和 14 天后的体重比较有显著差异(P≤0.05),第 1 组大鼠体重增加。所有治疗组(2、3、4 和 5)的大鼠体重都有所下降。分离物的抗生素耐药性检测模式显示出对所有抗生素的多重耐药性,其中耐药性最强的分离物是芽孢杆菌和肠杆菌。分离物的分子特征和鉴定结果表明,这些分离物与 arlettae 葡萄球菌、泄殖腔肠杆菌、癌原肠杆菌、诺索卡米尔氏不动杆菌、睾丸酵母菌、克雷伯氏菌、苏云金芽孢杆菌、枯草芽孢杆菌和蜡样芽孢杆菌亲缘关系密切,这表明在分离物的鉴定和特征描述方面,分子技术比对流技术更具特异性和准确性。此外,用茶叶处理的大鼠的肠道细菌数量明显增加,这表明茶叶有可能影响肠道细菌的数量和多样性。从本研究中可以看出,肠道细菌的分子特性鉴定可以确定细菌的种类水平,而且它们会受到饮食的影响。
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