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Background <p>Environmental wastewater is increasingly recognized as a critical reservoir and dissemination pathway for antimicrobial resistance (AMR). Extended-spectrum β-lactamase (ESBL)-producing Enterobacterales represent a major public health threat owing to their multidrug resistance and ability to disseminate resistance determinants through horizontal gene transfer. This study investigated the molecular epidemiology of multidrug-resistant ESBL-producing Enterobacterales recovered from hospital wastewater (HWW) and environmental wastewater (EWW) in Faisalabad, Pakistan.</p> Methods <p>A total of 100 samples from HWW and EWW sources were collected across Faisalabad. Enterobacterales were isolated using culture media and confirmed phenotypically. Antimicrobial susceptibility testing was performed using Kirby Bauer disc diffusion method, while ESBL production was confirmed by the combined disk diffusion test. Molecular detection of ESBL genes was conducted using multiplex PCR.</p> Results <p>Overall, 165 Enterobacterales isolates were recovered, including 86 (52.1%) from HWW and 79 (47.8%) from EWW. <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> were the predominant species in both wastewater sources. High resistance rates were observed against beta-lactam antibiotics except carbapenems. Overall, <i>bla</i><sub>TEM</sub> was the most prevalent β-lactamase gene, detected in 56/84 (66.7%) isolates, followed by <i>bla</i><sub>CTX-M</sub> in 50/84 (59.5%) and <i>bla</i><sub>SHV</sub> in 14/84 (16.7%) isolates. Among HWW isolates, <i>bla</i><sub>TEM</sub> was detected in 30/39 (76.9%), <i>bla</i><sub>CTX-M</sub> in 22/39 (56.4%), and <i>bla</i><sub>SHV</sub> in 3/39 (7.7%) isolates. In contrast, among EWW isolates, <i>bla</i><sub>CTX-M</sub> was detected in 28/45 (62.2%), <i>bla</i><sub>TEM</sub> in 26/45 (57.8%), and <i>bla</i><sub>SHV</sub> in 11/45 (24.4%) isolates.</p> Conclusions <p>The predominance of <i>bla</i><sub>CTX-M</sub> and <i>bla</i><sub>TEM</sub> underscores the environmental dissemination of clinically important resistance genes and the need for One Health surveillance.</p>

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High prevalence of ESBL encoding genes in enterobacterales isolated from hospital and environmental wastewater in Pakistan- One Health challenge

  • 期刊论文

  • 发表日期：

    2026年7月31日

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  • Muhammad Usman Qamar,
  • Muhammad Rizwan,
  • Fatima Tuz Zahra,
  • Faiqa Arshad,
  • Asma Ejaz,
  • Aatika,
  • Sana Rehman,
  • Zikria Saleem

摘要

Background

Environmental wastewater is increasingly recognized as a critical reservoir and dissemination pathway for antimicrobial resistance (AMR). Extended-spectrum β-lactamase (ESBL)-producing Enterobacterales represent a major public health threat owing to their multidrug resistance and ability to disseminate resistance determinants through horizontal gene transfer. This study investigated the molecular epidemiology of multidrug-resistant ESBL-producing Enterobacterales recovered from hospital wastewater (HWW) and environmental wastewater (EWW) in Faisalabad, Pakistan.

Methods

A total of 100 samples from HWW and EWW sources were collected across Faisalabad. Enterobacterales were isolated using culture media and confirmed phenotypically. Antimicrobial susceptibility testing was performed using Kirby Bauer disc diffusion method, while ESBL production was confirmed by the combined disk diffusion test. Molecular detection of ESBL genes was conducted using multiplex PCR.

Results

Overall, 165 Enterobacterales isolates were recovered, including 86 (52.1%) from HWW and 79 (47.8%) from EWW. Escherichia coli and Klebsiella pneumoniae were the predominant species in both wastewater sources. High resistance rates were observed against beta-lactam antibiotics except carbapenems. Overall, blaTEM was the most prevalent β-lactamase gene, detected in 56/84 (66.7%) isolates, followed by blaCTX-M in 50/84 (59.5%) and blaSHV in 14/84 (16.7%) isolates. Among HWW isolates, blaTEM was detected in 30/39 (76.9%), blaCTX-M in 22/39 (56.4%), and blaSHV in 3/39 (7.7%) isolates. In contrast, among EWW isolates, blaCTX-M was detected in 28/45 (62.2%), blaTEM in 26/45 (57.8%), and blaSHV in 11/45 (24.4%) isolates.

Conclusions

The predominance of blaCTX-M and blaTEM underscores the environmental dissemination of clinically important resistance genes and the need for One Health surveillance.