Microbial associations for pneumonia causative agents and level of their resistance to antimicrobial drugs during a new coronavirus infection pandemic

O. N. Kolotova, L. V. Kataeva, A. A. Vakarina, T. F. Stepanova, K. B. Stepanova
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Abstract

Introduction. Bacterial coinfection and secondary bacterial infection are considered critical risk factors for the severity and mortality of SARS-CoV-2-caused pneumonia. The aim of the study was to analyze a pattern of microbial associations between K. pneumoniae and A. baumannii isolated from the lower respiratory tract discharge and sectional material (lung tissue) of patients diagnosed with pneumonia, and to compare resistance level in monoculture and associations during new coronavirus infection pandemic. Materials and methods. A bacteriological study of 2689 sputum and bronchial washing samples from patients at infectious diseases hospitals, and 1411 lung pathological material samples was carried out. Bacterial isolates were identified by mass spectrometry. Antibiotic sensitivity for isolates was determined by the disk diffusion method. Genetic determinants of resistance to beta-lactam antibiotics were detected by PCR. Statistical data processing was performed using SPSS version 22 software. Results. K. pneumoniae and A. baumannii isolates were predominantly found in two- and three-pathogen associations. It was established that the resistance level of K. pneumoniae isolates in association with A. baumannii is significantly higher compared to that in monoculture for all antimicrobial drugs studied. At the same time, K. pneumoniae in combination with Candida spp. vs monoculture showed significantly lower level of resistance to ciprofloxacin, amikacin, cefotaxime, ceftazidime and amoxicillin/clavulanic acid. K. pneumoniae isolates carried resistance determinants to extended-spectrum beta-lactamases: OXA-48 — (22.5%), OXA-51 — (5.6%), OXA-23 — (4.2%), KPC — 70.9%, NDM — 7%. Of these, 14.1% of strains had the ability to co-produce serine carbapenemases OXA-48 and KPC. Sputum and lung tissue A. baumannii isolates exhibited extremely high multiple resistance regardless of their associations with other microorganisms. Microbiome species similarity in the lower respiratory tract and lung tissue discharge was revealed. The proportion of lung tissue vs sputum resistant strains of K. pneumoniae and A. baumannii was significantly higher. Conclusion. The detection of of multiple drug resistant K. pneumoniae and A. baumannii isolates as well as their associations may indicate aggravated pneumonia severity.
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新冠状病毒感染大流行期间肺炎致病菌的微生物关联及其对抗菌药物的耐药性水平
导言细菌合并感染和继发性细菌感染被认为是导致 SARS-CoV-2 引起的肺炎严重程度和死亡率的关键风险因素。本研究旨在分析从确诊为肺炎患者的下呼吸道分泌物和切片材料(肺组织)中分离出的肺炎克氏菌和鲍曼不动杆菌之间的微生物关联模式,并比较在新冠状病毒感染大流行期间单株培养和关联培养的耐药性水平。材料和方法对传染病医院 2689 份患者痰液和支气管冲洗样本以及 1411 份肺部病理材料样本进行了细菌学研究。细菌分离物通过质谱法进行鉴定。用盘式扩散法测定分离菌的抗生素敏感性。通过 PCR 检测对β-内酰胺类抗生素产生耐药性的基因决定因素。统计数据处理采用 SPSS 22 版软件。结果肺炎克氏菌和鲍曼不动杆菌主要存在于两种病原体和三种病原体的结合体中。研究发现,肺炎克雷伯菌与鲍曼不动杆菌结合后,其对所有抗菌药物的耐药性水平都明显高于单一培养的情况。同时,肺炎克雷伯菌与念珠菌属结合与单一培养相比,对环丙沙星、阿米卡星、头孢他啶、头孢唑肟和阿莫西林/克拉维酸的耐药性水平明显较低。肺炎克雷伯菌分离物对广谱β-内酰胺酶具有耐药性:oxa-48-(22.5%)、oxa-51-(5.6%)、oxa-23-(4.2%)、kpc-70.9%、ndm-7%。其中,14.1%的菌株能够同时产生丝氨酸碳青霉烯酶 OXA-48 和 KPC。痰和肺组织中分离出的鲍曼不动杆菌表现出极高的多重耐药性,无论它们是否与其他微生物有关联。下呼吸道和肺组织分泌物中的微生物组物种具有相似性。肺组织与痰中肺炎克氏菌和鲍曼不动杆菌耐药菌株的比例明显更高。结论检出多重耐药的肺炎克氏菌和鲍曼不动杆菌以及它们之间的关联可能预示着肺炎的严重程度。
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