Sundaramoorthy Niranjana Sri, Thothathri Subramaniam, Bhaskaran Muthumeenakshi, GaneshPrasad ArunKumar, Nagarajan Saisubramanian
Center for Research in Infectious Diseases (CRID), School of Chemical and Biotechnology, SASTRA Deemed University, Tamil Nadu, Thanjavur, 613401, India.
AMB Express. 2021 Feb 15;11(1):27. doi: 10.1186/s13568-021-01181-0.
Bacteriophages are a promising alternative for curtailing infections caused by multi drug resistant (MDR) bacteria. The objective of the present study is to evaluate phage populations from water bodies to inhibit planktonic and biofilm mode of growth of drug resistant Klebsiella pneumoniae in vitro and curtail planktonic growth in vivo in a zebrafish model. Phage specific to K. pneumoniae (MTCC 432) was isolated from Ganges River (designated as KpG). One-step growth curve, in vitro time kill curve study and in vivo infection model were performed to evaluate the ability of phage to curtail planktonic growth. Crystal violet assay and colony biofilm assay were performed to determine the action of phages on biofilms. KpG phages had a greater burst size, better bactericidal potential and enhanced inhibitory effect against biofilms formed at liquid air and solid air interfaces. In vitro time kill assay showed a 3 log decline and a 6 log decline in K. pneumoniae colony counts, when phages were administered individually and in combination with streptomycin, respectively. In vivo injection of KpG phages revealed that it did not pose any toxicity to zebrafish as evidenced by liver/brain enzyme profiles and by histopathological analysis. The muscle tissue of zebrafish, infected with K. pneumoniae and treated with KpG phages alone and in combination with streptomycin showed a significant 77.7% and 97.2% decline in CFU/ml, respectively, relative to untreated control. Our study reveals that KpG phages has the potential to curtail plantonic and biofilm mode of growth in higher animal models.
噬菌体是抑制多重耐药（MDR）细菌引起的感染的一种有前景的替代方法。本研究的目的是评估水体中的噬菌体群体，以在体外抑制耐药肺炎克雷伯菌的浮游生长和生物膜生长模式，并在斑马鱼模型中抑制体内的浮游生长。从恒河分离出肺炎克雷伯菌（MTCC 432）特异性噬菌体（命名为KpG）。进行一步生长曲线、体外时间杀灭曲线研究和体内感染模型，以评估噬菌体抑制浮游生长的能力。进行结晶紫测定和菌落生物膜测定，以确定噬菌体对生物膜的作用。KpG噬菌体具有更大的裂解量、更好的杀菌潜力，并且对在液气和固气界面形成的生物膜具有增强的抑制作用。体外时间杀灭试验表明，单独施用噬菌体和与链霉素联合施用时，肺炎克雷伯菌菌落计数分别下降3个对数和6个对数。体内注射KpG噬菌体表明，根据肝/脑酶谱和组织病理学分析，它对斑马鱼没有任何毒性。感染肺炎克雷伯菌并单独用KpG噬菌体以及与链霉素联合处理的斑马鱼肌肉组织，相对于未处理的对照，CFU/ml分别显著下降77.7%和97.2%。我们的研究表明，KpG噬菌体有潜力在高等动物模型中抑制浮游生长和生物膜生长模式。
