Lactiker Research Group, Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain; Bioaraba Health Research Institute-Prevention, Promotion and Health Care, 01009 Vitoria-Gasteiz, Spain; Joint Research Laboratory on Environmental Antibiotic Resistance, Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain.
Lactiker Research Group, Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain; Bioaraba Health Research Institute-Prevention, Promotion and Health Care, 01009 Vitoria-Gasteiz, Spain; Joint Research Laboratory on Environmental Antibiotic Resistance, Department of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), Paseo de la Universidad 7, 01006 Vitoria-Gasteiz, Spain.
Food Res Int. 2024 Jul;187:114308. doi: 10.1016/j.foodres.2024.114308. Epub 2024 Apr 18.
Antimicrobial resistance (AMR) is a significant public health threat, with the food production chain, and, specifically, fermented products, as a potential vehicle for dissemination. However, information about dairy products, especially raw ewe milk cheeses, is limited. The present study analysed, for the first time, the occurrence of AMRs related to lactic acid bacteria (LAB) along a raw ewe milk cheese production chain for the most common antimicrobial agents used on farms (dihydrostreptomycin, benzylpenicillin, amoxicillin and polymyxin B). More than 200 LAB isolates were obtained and identified by Sanger sequencing (V1-V3 16S rRNA regions); these isolates included 8 LAB genera and 21 species. Significant differences in LAB composition were observed throughout the production chain (P ≤ 0.001), with Enterococcus (e.g., E. hirae and E. faecalis) and Bacillus (e.g., B. thuringiensis and B. cereus) predominating in ovine faeces and raw ewe milk, respectively, along with Lactococcus (L. lactis) in whey and fresh cheeses, while Lactobacillus and Lacticaseibacillus species (e.g., Lactobacillus sp. and L. paracasei) prevailed in ripened cheeses. Phenotypically, by broth microdilution, Lactococcus, Enterococcus and Bacillus species presented the greatest resistance rates (on average, 78.2 %, 56.8 % and 53.4 %, respectively), specifically against polymyxin B, and were more susceptible to dihydrostreptomycin. Conversely, Lacticaseibacillus and Lactobacillus were more susceptible to all antimicrobials tested (31.4 % and 39.1 %, respectively). Thus, resistance patterns and multidrug resistance were reduced along the production chain (P ≤ 0.05). Genotypically, through HT-qPCR, 31 antimicrobial resistance genes (ARGs) and 6 mobile genetic elements (MGEs) were detected, predominating Str, StrB and aadA-01, related to aminoglycoside resistance, and the transposons tnpA-02 and tnpA-01. In general, a significant reduction in ARGs and MGEs abundances was also observed throughout the production chain (P ≤ 0.001). The current findings indicate that LAB dynamics throughout the raw ewe milk cheese production chain facilitated a reduction in AMRs, which has not been reported to date.
抗生素耐药性（AMR）是一个重大的公共卫生威胁，食品生产链，特别是发酵产品，是传播的潜在载体。然而，关于乳制品的信息，特别是生羊奶奶酪，是有限的。本研究首次分析了与乳酸细菌（LAB）相关的 AMR 沿生羊奶奶酪生产链的发生情况，用于农场最常见的抗生素（二氢链霉素、苄青霉素、阿莫西林和多粘菌素 B）。通过 Sanger 测序（V1-V3 16S rRNA 区）获得并鉴定了 200 多个 LAB 分离株；这些分离株包括 8 个 LAB 属和 21 个种。在整个生产链中观察到 LAB 组成的显著差异（P≤0.001），肠球菌（如屎肠球菌和粪肠球菌）和芽孢杆菌（如苏云金芽孢杆菌和蜡样芽孢杆菌）分别在绵羊粪便和生羊奶中占优势，而乳球菌（如乳球菌）在乳清和新鲜奶酪中占优势，而乳杆菌和乳酸乳球菌（如乳杆菌和副干酪乳杆菌）在成熟奶酪中占优势。表型上，通过肉汤微量稀释法，乳球菌、肠球菌和芽孢杆菌的耐药率最高（平均分别为 78.2%、56.8%和 53.4%），特别是对多粘菌素 B，而对二氢链霉素更敏感。相反，乳酸乳球菌和乳杆菌对所有测试的抗生素更敏感（分别为 31.4%和 39.1%）。因此，耐药模式和多药耐药性沿生产链减少（P≤0.05）。通过 HT-qPCR 检测到 31 种抗生素耐药基因（ARGs）和 6 种移动遗传元件（MGEs），以 Str、StrB 和 aadA-01 为主，与氨基糖苷类耐药性有关，以及转座子 tnpA-02 和 tnpA-01。一般来说，整个生产链中 ARGs 和 MGEs 的丰度也显著降低（P≤0.001）。目前的研究结果表明，生羊奶奶酪生产链中 LAB 动态的变化促进了 AMR 的减少，这在以前的报道中尚未发现。
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