Bakon Sophia Karen, Mohamad Zuraifah Asrah, Jamilan Mohd Azerulazree, Hashim Hazimah, Kuman Mohamed Yazid, Shaharudin Rafiza, Ahmad Norazah, Muhamad Nor Asiah
Environmental Health Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam, Malaysia.
Nutritional, Metabolic and Cardiovascular Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health Malaysia, Shah Alam, Malaysia.
JMIR Res Protoc. 2023 May 29;12:e39022. doi: 10.2196/39022.
Antimicrobial resistance (AMR) has emerged as a major global public health challenge due to the overuse and misuse of antibiotics for humans and animals. Hospitals are among the major users of antibiotics, thereby having a large contribution to AMR.
The aim of this study is to determine the prevalence of antibiotic-resistant pathogenic bacteria and the level of antibiotic residues in the hospital effluents in Selangor, Malaysia.
A cross-sectional study will be performed in the state of Selangor, Malaysia. Tertiary hospitals will be identified based on the inclusion and exclusion criteria. The methods are divided into three phases: sample collection, microbiological analysis, and chemical analysis. Microbiological analyses will include the isolation of bacteria from hospital effluents by culturing on selective media. Antibiotic sensitivity testing will be performed on the isolated bacteria against ceftriaxone, ciprofloxacin, meropenem, vancomycin, colistin, and piperacillin/tazobactam. The identification of bacteria will be confirmed using 16S RNA polymerase chain reaction (PCR) and multiplex PCR will be performed to detect resistance genes (ermB, mecA, bla, bla, bla, bla, VanA, VanB, VanC1, mcr-1, mcr-2, mcr-3, Intl1, Intl2, and qnrA). Finally, the level of antibiotic residues will be measured using ultrahigh-performance liquid chromatography.
The expected outcomes will be the prevalence of antibiotic-resistant Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter (ESKAPE) bacterial species from the hospital effluents, the occurrence of antibiotic resistance genes (ARGs) from the isolated ESKAPE bacteria, and the level of antibiotic residues that may be detected from the effluent. Sampling has been conducted in three hospitals. Data analysis from one hospital showed that as of July 2022, 80% (8/10) of E. faecium isolates were resistant to vancomycin and 10% (1/10) were resistant to ciprofloxacin. Further analysis will be conducted to determine if the isolates harbor any ARGs and effluent samples are being analyzed to detect antibiotic residues. Sampling activities will be resumed after being suspended due to the COVID-19 pandemic and are scheduled to end by December 2022.
This study will provide the first baseline information to elucidate the current status of AMR of highly pathogenic bacteria present in hospital effluents in Malaysia.
INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/39022.
由于人类和动物对抗生素的过度使用和滥用，抗菌药物耐药性（AMR）已成为全球主要的公共卫生挑战。医院是抗生素的主要使用者之一，因此对AMR有很大的影响。
本研究的目的是确定马来西亚雪兰莪医院污水中抗生素耐药病原菌的流行情况以及抗生素残留水平。
将在马来西亚雪兰莪州进行一项横断面研究。将根据纳入和排除标准确定三级医院。方法分为三个阶段：样本采集、微生物分析和化学分析。微生物分析将包括通过在选择性培养基上培养从医院污水中分离细菌。将对分离出的细菌进行头孢曲松、环丙沙星、美罗培南、万古霉素、黏菌素和哌拉西林/他唑巴坦的药敏试验。将使用16S RNA聚合酶链反应（PCR）确认细菌的鉴定，并进行多重PCR以检测耐药基因（ermB、mecA、bla、bla、bla、bla、VanA、VanB、VanC1、mcr-1、mcr-2、mcr-3、Intl1、Intl2和qnrA）。最后，将使用超高效液相色谱法测量抗生素残留水平。
预期结果将是医院污水中耐抗生素的粪肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌（ESKAPE）细菌种类的流行情况，分离出的ESKAPE细菌中抗生素耐药基因（ARGs）的出现情况，以及可能从污水中检测到的抗生素残留水平。已在三家医院进行了采样。一家医院的数据分析表明，截至2022年7月，80%（8/10）的粪肠球菌分离株对万古霉素耐药，10%（1/10）对环丙沙星耐药。将进行进一步分析以确定分离株是否携带任何ARGs，并正在分析污水样本以检测抗生素残留。由于COVID-19大流行而暂停的采样活动将恢复，并计划于2022年12月结束。
本研究将提供首个基线信息，以阐明马来西亚医院污水中高致病性细菌的AMR现状。
国际注册报告标识符（IRRID）：DERR1-10.2196/39022。
