Antimicrobial Resistance Gene Patterns in Traditional Montenegrin Njeguški Cheese Revealed by qPCR

<b>Background/Objectives</b>: This study was aimed to investigate the safety profile of traditional Montenegrin <i>Njeguški</i> cheese by quantifying genes associated with resistance to clinically important antibiotics. <b>Methods</b>: Samples of <i>Njeguški</i> cheese were sourced from three artisan producers in Montenegro, identified as A, B, and C, with three individual batches selected per producer. Quantitative PCR (qPCR) was performed on bacterial DNA extracted directly from samples to detect genes encoding resistance to macrolide-lincosamide-streptogramin B (MLS_B) [<i>erm</i>(A), <i>erm</i>(B), <i>erm</i>(C)], vancomycin (<i>van</i>A, <i>van</i>B), tetracyclines [<i>tet</i>(M), <i>tet</i>(O), <i>tet</i>(S), <i>tet</i>(K), <i>tet</i>(W)], β-lactams (<i>mecA</i>, <i>blaZ</i>), aminoglycosides [<i>aac (6')-Ie aph (2″)-Ia</i>], and carbapenems (<i>bla</i>_KPC, <i>bla</i>_OXA-48, <i>bla</i>_NDM-1, <i>bla</i>_GES, and <i>bla</i>_VIM). <b>Results</b>: Among the MLS_B resistance genes, <i>erm</i>(B) was detected in all samples, <i>erm</i>(C) was present only in those from producer B, while <i>erm</i>(A) was found exclusively in batch 3 from producer C. Tetracycline resistance genes were widely distributed, except for <i>tet</i>(O), which was absent in batch 3 from producers A and B. Regarding β-lactam resistance, both <i>blaZ</i> and <i>mecA</i> were consistently detected across all samples, with statistically significant differences observed between producers. None of the samples tested positive for vancomycin resistance genes or the aminoglycoside resistance gene, regardless of producer. Among the carbapenemase genes analyzed, <i>bla</i>_NDM-1 was the only one detected, found in most samples from producers B and C. <b>Conclusions</b>: This research provides the first risk assessment of artisanal and commercial <i>Njeguški</i> cheese regarding antimicrobial resistance genes. The findings offer valuable insights to enhance the microbiological safety of traditional Montenegrin cheeses, supporting consumer confidence in local and international markets.