Frontiers in microbiology · 2026
Concordance between environmental resistomes and pathogenic phenotypes: a case study of multidrug-resistant Klebsiella pneumoniae in a drinking water source in Guizhou, China.
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Qian Wanning, Han Ao, Al Hatmi Abdullah M S, Wang Yanyan, Rafiq Muhammad, Cui Guzhen, Zhou Shaoqin, Li Shijun, Kang Yingqian
Аннотация
INTRODUCTION: The persistence of antibiotic resistance in aquatic environments poses a public health concern, particularly when drinking water sources act as reservoirs for multidrug-resistant opportunistic pathogens. However, the linkage between environmental resistomes and the resistance phenotypes of cultivable bacteria remains under-characterized. This case study investigated this relationship in a karst drinking water source in Guizhou, China.
METHODS: Surface water samples from seven sites were analyzed for antibiotic residues using LC-MS/MS. Metagenomic sequencing was conducted on selected contamination hotspots to characterize microbial communities and antibiotic resistance genes (ARGs). Cultivable bacteria were isolated, identified via 16S rRNA sequencing, and tested for antimicrobial susceptibility. To validate resistance mechanisms, a multidrug-resistant Klebsiella pneumoniae isolate was analyzed for tetA expression using RT-qPCR.
RESULTS: Antibiotic residues were detected across all sites, with sulfonamides and tetracyclines being the most prevalent. Consistent with this chemical pressure, metagenomic analysis identified corresponding ARGs, including sul1 and tet(Q), which functionally clustered with mobile genetic elements. From the contaminated matrix (sample W2), a multidrug-resistant Klebsiella pneumoniae strain (B8) was recovered. Mechanistic validation revealed a 2.78-fold upregulation of the tetA efflux pump gene in this strain.
DISCUSSION: These findings demonstrate a concordance among chemical selection pressures, environmental resistomes, and active resistance phenotypes. The results indicate that drinking water sources can harbor and maintain clinically relevant resistant bacteria, supporting the implementation of integrated surveillance strategies to evaluate biological risks.
METHODS: Surface water samples from seven sites were analyzed for antibiotic residues using LC-MS/MS. Metagenomic sequencing was conducted on selected contamination hotspots to characterize microbial communities and antibiotic resistance genes (ARGs). Cultivable bacteria were isolated, identified via 16S rRNA sequencing, and tested for antimicrobial susceptibility. To validate resistance mechanisms, a multidrug-resistant Klebsiella pneumoniae isolate was analyzed for tetA expression using RT-qPCR.
RESULTS: Antibiotic residues were detected across all sites, with sulfonamides and tetracyclines being the most prevalent. Consistent with this chemical pressure, metagenomic analysis identified corresponding ARGs, including sul1 and tet(Q), which functionally clustered with mobile genetic elements. From the contaminated matrix (sample W2), a multidrug-resistant Klebsiella pneumoniae strain (B8) was recovered. Mechanistic validation revealed a 2.78-fold upregulation of the tetA efflux pump gene in this strain.
DISCUSSION: These findings demonstrate a concordance among chemical selection pressures, environmental resistomes, and active resistance phenotypes. The results indicate that drinking water sources can harbor and maintain clinically relevant resistant bacteria, supporting the implementation of integrated surveillance strategies to evaluate biological risks.