Microbiology and Biotechnology Research laboratory, Department of Biotechnology, Fatima Jinnah Women University, Rawalpindi, Pakistan.
Department of Orthopaedic Surgery, Fauji Foundation Hospital, Rawalpindi, Pakistan.
PLoS One. 2023 Oct 17;18(10):e0292956. doi: 10.1371/journal.pone.0292956. eCollection 2023.
The exponential increase in the prevalence of multidrug resistant bacteria has resulted in limiting surgical treatment options globally, potentially causing biofilm-related complications, implant failure, and severe consequences. This study aims to isolate and characterize bacteria from post-surgical orthopaedic implant infections and screening for multiple antibiotic resistance. A cross-sectional study was conducted, involving isolation of forty-four dominant pathogenic bacterial isolates from 16 infected implant samples from across Islamabad and Rawalpindi. Out of forty-four, 38% cocci and 61% bacilli were obtained. Approximately 90% of isolates showed multiple antibiotic resistance (MAR) index of more than 0.2. Eleven strains were identified via 16S rRNA gene sequencing as Pseudomonas aeruginosa, Bacillus spp., Planococcus chinensis, Staphylococcus, Escherichia coli and Enterobacter cloacae. The bacterial strain E. coli MB641 showed sensitivity to Polymyxin only, and was resistant to all other antibiotics used. Maximum biofilm forming ability 0.532 ± 0.06, 0.55 ± 0.01 and 0.557 ± 0.07 was observed in Pseudomonas aeruginosa MB663, Pseudomonas aeruginosa MB664 and Bacillus spp. MB647 respectively after 24 hours of incubation. EPS production of bacterial strains was assessed, the polysaccharides and protein content of EPS were found to be in the range of 11-32 μg/ml and 2-10 μg/ml, respectively. Fourier transform infrared spectroscopic analysis of EPS showed the presence of carbohydrates, proteins, alkyl halides, and nucleic acids. X-ray diffraction analysis revealed crystalline structure of EPS extracted from biofilm forming bacteria. These findings suggest a high prevalence of antibiotic-resistant bacteria in orthopaedic implant-associated surgeries, highlighting the urgent need for ongoing monitoring and microorganism testing in infected implants.
耐多药细菌的患病率呈指数级增长,这导致了全球范围内手术治疗选择的受限,可能导致生物膜相关并发症、植入物失败和严重后果。本研究旨在从术后骨科植入物感染中分离和鉴定细菌,并进行多重抗生素耐药性筛查。这是一项横断面研究,涉及从伊斯兰堡和拉瓦尔品第的 16 个感染植入物样本中分离出 44 株优势致病细菌分离株。在这 44 株中,球菌占 38%,杆菌占 61%。大约 90%的分离株显示出超过 0.2 的多重抗生素耐药(MAR)指数。通过 16S rRNA 基因测序,有 11 株被鉴定为铜绿假单胞菌、芽孢杆菌、菜氏栖菜球菌、葡萄球菌、大肠杆菌和阴沟肠杆菌。细菌株 E. coli MB641 仅对多粘菌素敏感,对所有其他使用的抗生素均有耐药性。在 24 小时孵育后,观察到 Pseudomonas aeruginosa MB663、Pseudomonas aeruginosa MB664 和 Bacillus spp. MB647 的最大生物膜形成能力分别为 0.532±0.06、0.55±0.01 和 0.557±0.07。评估了细菌株的 EPS 产生情况,发现 EPS 的多糖和蛋白质含量分别在 11-32μg/ml 和 2-10μg/ml 范围内。EPS 的傅里叶变换红外光谱分析显示存在碳水化合物、蛋白质、烷基卤化物和核酸。从形成生物膜的细菌中提取的 EPS 的 X 射线衍射分析显示出结晶结构。这些发现表明骨科植入物相关手术中存在高比例的抗生素耐药细菌,强调了对感染植入物进行持续监测和微生物检测的迫切需要。