Department of Medical Microbiology, School of Biomedical Sciences, Makerere University College of Health Sciences, Kampala, Uganda.
Department of Immunology and Molecular Biology, School of Biomedical Sciences, Makerere University College of Health Sciences, Kampala, Uganda.
PLoS One. 2023 Jun 8;18(6):e0286955. doi: 10.1371/journal.pone.0286955. eCollection 2023.
Escherichia coli, Klebsiella pneumoniae and Enterobacter (EKE) are the leading cause of mortality and morbidity in neonates in Africa. The management of EKE infections remains challenging given the global emergence of carbapenem resistance in Gram-negative bacteria. This study aimed to investigate the source of EKE organisms for neonates in the maternity environment of a national referral hospital in Uganda, by examining the phenotypic and molecular characteristics of isolates from mothers, neonates, and maternity ward.
From August 2015 to August 2016, we conducted a cross-sectional study of pregnant women admitted for elective surgical delivery at Mulago hospital in Kampala, Uganda; we sampled (nose, armpit, groin) 137 pregnant women and their newborns (n = 137), as well as health workers (n = 67) and inanimate objects (n = 70 -beds, ventilator tubes, sinks, toilets, door-handles) in the maternity ward. Samples (swabs) were cultured for growth of EKE bacteria and isolates phenotypically/molecularly investigated for antibiotic sensitivity, as well as β-lactamase and carbapenemase activity. To infer relationships among the EKE isolates, spatial cluster analysis of phenotypic and genotypic susceptibility characteristics was done using the Ridom server.
Gram-negative bacteria were isolated from 21 mothers (15%), 15 neonates (11%), 2 health workers (3%), and 13 inanimate objects (19%); a total of 131 Gram-negative isolates were identified of which 104 were EKE bacteria i.e., 23 (22%) E. coli, 50 (48%) K. pneumoniae, and 31 (30%) Enterobacter. Carbapenems were the most effective antibiotics as 89% (93/104) of the isolates were susceptible to meropenem; however, multidrug resistance was prevalent i.e., 61% (63/104). Furthermore, carbapenemase production and carbapenemase gene prevalence were low; 10% (10/104) and 6% (6/104), respectively. Extended spectrum β-lactamase (ESBL) production occurred in 37 (36%) isolates though 61 (59%) carried ESBL-encoding genes, mainly blaCTX-M (93%, 57/61) implying that blaCTX-M is the ideal gene for tracking ESBL-mediated resistance at Mulago. Additionally, spatial cluster analysis revealed isolates from mothers, new-borns, health workers, and environment with similar phenotypic/genotypic characteristics, suggesting transmission of multidrug-resistant EKE to new-borns.
Our study shows evidence of transmission of drug resistant EKE bacteria in the maternity ward of Mulago hospital, and the dynamics in the ward are more likely to be responsible for transmission but not individual mother characteristics. The high prevalence of drug resistance genes highlights the need for more effective infection prevention/control measures and antimicrobial stewardship programs to reduce spread of drug-resistant bacteria in the hospital, and improve patient outcomes.
产酸克雷伯菌、大肠埃希菌和肠杆菌(EKE)是非洲新生儿死亡率和发病率的主要原因。鉴于全球革兰氏阴性菌中碳青霉烯类耐药性的出现,EKE 感染的治疗仍然具有挑战性。本研究旨在通过检查母亲、新生儿和产房的分离株的表型和分子特征,调查乌干达一家国家转诊医院产妇环境中 EKE 生物体的来源。
2015 年 8 月至 2016 年 8 月,我们对乌干达坎帕拉穆拉戈医院接受择期手术分娩的孕妇进行了一项横断面研究;我们对 137 名孕妇及其新生儿(n=137)、67 名卫生工作者(n=67)和 70 个无生命物体(床、呼吸机管、水槽、厕所、门把手)进行了采样(鼻拭子、腋窝、腹股沟)。对样本(拭子)进行了 EKE 细菌生长培养,并对分离株进行了表型/分子抗生素敏感性、β-内酰胺酶和碳青霉烯酶活性检测。为了推断 EKE 分离株之间的关系,我们使用 Ridom 服务器对表型和基因型敏感性特征进行了空间聚类分析。
从 21 名母亲(15%)、15 名新生儿(11%)、2 名卫生工作者(3%)和 13 个无生命物体(19%)中分离出革兰氏阴性菌;共鉴定出 131 株革兰氏阴性菌,其中 104 株为 EKE 细菌,即 23 株(22%)大肠埃希菌、50 株(48%)肺炎克雷伯菌和 31 株(30%)肠杆菌。碳青霉烯类抗生素是最有效的抗生素,因为 89%(93/104)的分离株对美罗培南敏感;然而,多药耐药性很常见,即 61%(63/104)。此外,碳青霉烯酶的产生和碳青霉烯酶基因的流行率较低,分别为 10%(10/104)和 6%(6/104)。37 株(36%)分离株产生了超广谱β-内酰胺酶(ESBL),但 61 株(59%)携带 ESBL 编码基因,主要是 blaCTX-M(93%,57/61),这表明 blaCTX-M 是追踪 Mulago 产 ESBL 介导耐药性的理想基因。此外,空间聚类分析显示,来自母亲、新生儿、卫生工作者和环境的分离株具有相似的表型/基因型特征,表明多药耐药 EKE 向新生儿的传播。
我们的研究表明,穆拉戈医院产房中存在耐药 EKE 细菌的传播证据,病房中的动态变化更有可能导致传播,而不是个体母亲的特征。耐药基因的高流行率突出表明,需要采取更有效的感染预防/控制措施和抗菌药物管理计划,以减少医院内耐药菌的传播,并改善患者的预后。