尼日利亚哈科特港市酸奶分离菌的分子特征及抗生素抗性基因检测

Amadi Lawrence Omunakwe, Okpara Joyce Chimuanya, Ugboma John Chukwuemeka
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摘要

目前,分子鉴定因其准确性和可靠性正在取代传统的方法,而抗菌治疗的有效性由于抗生素耐药性而不断下降。本研究旨在鉴定分离的细菌;对从酸奶样品中分离得到的菌株进行药敏试验后进行表型和分子检测以及抗性基因检测。标准微生物技术和分子分析应用于两种样品(商业和自制酸奶)进行物种验证。共鉴定出44种细菌,表型上分属于3属;从分子分析中发现了芽孢杆菌、葡萄球菌、乳酸杆菌和双歧杆菌属。酸奶样品的微生物负荷差异无统计学意义(p≥0.05)。采用Kirby-Bauer圆盘扩散法对几种标准抗生素进行敏感性试验。结果显示,芽孢杆菌和葡萄球菌对氨苄西林和扩增素耐药(100%),对氧氟沙星和庆大霉素敏感。乳酸菌对氧氟沙星和头孢他啶敏感(100%),对氨苄西林、增敏素和环丙沙星耐药(100%)。6种耐药菌株分别为金黄色葡萄球菌CP019117、表皮葡萄球菌AB68833、巨型芽孢杆菌KC246043、蜡样芽孢杆菌NC004722、干酪乳杆菌NC008526和乳酸双歧杆菌CP003941。携带mecA基因的耐药菌为金黄色葡萄球菌和表皮葡萄球菌,携带ampC基因的耐药菌为乳酸双歧杆菌和干酪乳杆菌。然而,在任何芽孢杆菌物种的基因组中都没有发现这两个基因。然而,数据还显示,自制酸奶样品中的细菌种类对mecA和ampC耐药基因呈阴性,而商品样品中的细菌种类呈阳性。这些基因导致了分离的细菌具有高水平的多药耐药(MDR)。商业酸奶样品中细菌种类中耐药基因的存在仍然是食品安全的一个挑战。因此,在此提倡良好的生产规范,适当的卫生和环境卫生,以避免严重的新出现的食源性疾病。
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Molecular characterization and detection of antibiotic resistant genes of bacteria isolated from yoghurt in Port Harcourt Metropolis, Nigeria
Currently, molecular identification is replacing the conventional method because of its precision and reliability whereas the effectiveness of antibacterial treatments has continuously declined due to antimicrobial resistance (AMR). This study aimed to identify the bacterial isolates; phenotypically and molecularly as well as detect the resistant genes after susceptibility testing of the isolates obtained from yoghurt samples. Standard microbiological techniques and molecular analysis were applied on both samples (commercial and home-made yoghurt) for species validation. Forty-four (44) bacterial species were identified, phenotypically belonging to three (3) genera; Bacillus, Staphylococcus and Lactobacillus and an additional genus Bifidobacterium emerged from molecular analysis. The microbial load of the yoghurt samples was not statistically significant at (p≥0.05). A sensitivity test on the species was carried out using Kirby-Bauer disc diffusion method with some standard antibiotics. The results revealed that Bacillus and Staphylococcus species were resistant to ampicillin and augmentin (100%) but susceptible to ofloxacin and gentamicin respectively. Lactobacillus spp. were susceptible to ofloxacin and ceftazidime (100%), and resistant to ampicillin, augmentin, and ciprofloxacin (100%). The six most resistant species were molecularly identified as S. aureus CP019117, S. epidermidis AB68833, B. megaterium KC246043, B. cereus NC004722, Lactobacillus casei NC008526 and Bifidobacterium lactis CP003941. Resistant bacteria with mecA gene are S. aureus and S. epidermidis and those with ampC gene are Bifidobacterium lactis and Lactobacillus casei. However, neither gene was found in the genome of any Bacillus species. However, the data also revealed that the bacterial species in home-made yoghurt samples were negative for mecA and ampC resistant genes but positive in the commercial samples. These genes contributed to the bacterial isolates' high levels of multidrug resistance (MDR). The presence of resistant genes in bacterial species from commercial yoghurt samples remains a challenge for food safety. Therefore, good manufacturing practice, proper hygiene and sanitation is hereby advocated to avoid serious emerging foodborne illnesses.
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