Pallavali Roja Rani, Degati Vijaya Lakshmi, Lomada Dakshayani, Reddy Madhava C, Durbaka Vijaya Raghava Prasad
Department of Microbiology, Yogi Vemana University, Kadapa, AP, India.
Department of Genetic and Genomics, Yogi Vemana University, Kadapa, AP, India.
PLoS One. 2017 Jul 18;12(7):e0179245. doi: 10.1371/journal.pone.0179245. eCollection 2017.
Multi-drug resistance has become a major problem for the treatment of pathogenic bacterial infections. The use of bacteriophages is an attractive approach to overcome the problem of drug resistance in several pathogens that cause fatal diseases. Our study aimed to isolate multi drug resistant bacteria from patients with septic wounds and then isolate and apply bacteriophages in vitro as alternative therapeutic agents. Pus samples were aseptically collected from Rajiv Gandhi Institute of Medical Science (RIMS), Kadapa, A.P., and samples were analyzed by gram staining, evaluating morphological characteristics, and biochemical methods. MDR-bacterial strains were collected using the Kirby-Bauer disk diffusion method against a variety of antibiotics. Bacteriophages were collected and tested in vitro for lytic activity against MDR-bacterial isolates. Analysis of the pus swab samples revealed that the most of the isolates detected had Pseudomonas aeruginosa as the predominant bacterium, followed by Staphylococcus aureus, Klebsiella pneumoniae and Escherichia coli. Our results suggested that gram-negative bacteria were more predominant than gram-positive bacteria in septic wounds; most of these isolates were resistant to ampicillin, amoxicillin, penicillin, vancomycin and tetracycline. All the gram-positive isolates (100%) were multi-drug resistant, whereas 86% of the gram-negative isolates had a drug resistant nature. Further bacteriophages isolated from sewage demonstrated perfect lytic activity against the multi-drug resistant bacteria causing septic wounds. In vitro analysis of the isolated bacteriophages demonstrated perfect lysis against the corresponding MDR-bacteria, and these isolated phages may be promising as a first choice for prophylaxis against wound sepsis, Moreover, phage therapy does not enhance multi-drug resistance in bacteria and could work simultaneously on a wide variety of MDR-bacteria when used in a bacteriophage cocktail. Hence, our results suggest that these bacteriophages could be potential therapeutic options for treating septic wounds caused by P. aeruginosa, S. aureus, K. pneumoniae and E. coli.
多重耐药已成为治疗致病性细菌感染的一个主要问题。使用噬菌体是一种有吸引力的方法，可用于克服几种导致致命疾病的病原体的耐药性问题。我们的研究旨在从脓毒症伤口患者中分离出多重耐药细菌，然后在体外分离并应用噬菌体作为替代治疗剂。从印度安得拉邦卡达帕市的拉吉夫·甘地医学科学研究所（RIMS）无菌采集脓液样本，并通过革兰氏染色、评估形态特征和生化方法对样本进行分析。使用针对多种抗生素的 Kirby-Bauer 纸片扩散法收集多重耐药细菌菌株。收集噬菌体并在体外测试其对多重耐药细菌分离株的裂解活性。对脓液拭子样本的分析表明，检测到的大多数分离株以铜绿假单胞菌为主，其次是金黄色葡萄球菌、肺炎克雷伯菌和大肠杆菌。我们的结果表明，脓毒症伤口中革兰氏阴性菌比革兰氏阳性菌更占优势；这些分离株大多对氨苄青霉素、阿莫西林、青霉素、万古霉素和四环素耐药。所有革兰氏阳性分离株（100%）均为多重耐药，而 86%的革兰氏阴性分离株具有耐药性。从污水中分离出的噬菌体对导致脓毒症伤口的多重耐药细菌表现出完美的裂解活性。对分离出的噬菌体进行体外分析表明，其对相应的多重耐药细菌具有完美的裂解作用，这些分离出的噬菌体有望作为预防伤口脓毒症的首选药物。此外，噬菌体疗法不会增强细菌的多重耐药性，当以噬菌体鸡尾酒形式使用时，可同时作用于多种多重耐药细菌。因此，我们的结果表明，这些噬菌体可能是治疗由铜绿假单胞菌、金黄色葡萄球菌、肺炎克雷伯菌和大肠杆菌引起的脓毒症伤口的潜在治疗选择。
