Distribution of mcr-1 Harboring Hypervirulent Klebsiella pneumoniae in Clinical Specimens and Lytic Activity of Bacteriophage KpnM Against Isolates

Bilal Aslam,1 Muhammad Hussnain Siddique,2 Abu Baker Siddique,1 Muhammad Shafique,1 Saima Muzammil,1 Mohsin Khurshid,1 Muhammad Hidayat Rasool,1 Moeed Ahmad,1 Tamoor Hamid Chaudhry,3 Afreenish Amir,3 Muhammad Salman,3 Zulqarnain Baloch,4 Norah A Alturki,5 Ahmad Alzamami6 1Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan; 2Department of Bioinformatics & Biotechnology, Government College University Faisalabad, Faisalabad, Pakistan; 3Public Health Laboratories Division, National Institute of Health, Islamabad, Pakistan; 4Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, People’s Republic of China; 5Clinical Laboratory Science Department, College of Applied Medical Science, King Saud University, Riyadh, Saudi Arabia; 6Clinical Laboratory Science Department, College of Applied Medical Science, Shaqra University, AlQuwayiyah, Saudi ArabiaCorrespondence: Bilal Aslam, Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan, Email drbilalaslam@gcuf.edu.pk Ahmad Alzamami, Clinical Laboratory Science Department, College of Applied Medical Science, Shaqra University, AlQuwayiyah, Saudi Arabia, Email aalzamami@su.edu.saBackground: The World Health Organization (WHO) has declared the multi-drug resistant (MDR) Klebsiella pneumoniae as one of the critical bacterial pathogens. The dearth of new antibiotics and inadequate therapeutic options necessitate finding alternative options. Bacteriophages are known as enemies of bacteria and are well-recognized to fight MDR pathogens.Methods: A total of 150 samples were collected from different clinical specimens through a convenient sampling technique. Isolation, identification, and antibiotic susceptibility testing (AST) of K. pneumoniae were done by standard and validated microbiological procedures. Molecular identification of virulence factors and antibiotic resistance genes (ARGs) was carried out through polymerase chain reaction (PCR) by using specific primers. For bacteriophage isolation, hospital sewage samples were processed for phage enrichment, purification, and further characterization ie, transmission electron microscopy (TEM) and stability testing, etc. followed by evaluation of the lytic potential of the phage.Results: Overall, a total of 41% of isolates of K. pneumoniae were observed as hypervirulent K. pneumoniae (hvKp). Among hvKp, a total of 12 (42%) were detected as MDR hvKp. A total of 37% of all MDR isolates were found resistant to colistin, and 66% of the colistin resistance isolates were recorded as mcr-1 positive. Isolated phage KpnM had shown lytic activity against 53 (79%) K. pneumoniae isolates. Remarkably, all 8 mcr-1 harboring MDR hvKp and non-hvKp isolates were susceptible to KpnM phage.Conclusion: Significant distribution of mcr-1 harboring hypervirulent Klebsiella pneumoniae was observed in clinical specimens, which is worrisome for the health system of the country. Characterized phage KpnM exhibited encouraging results and showed the lytic activity against the mcr-1 harboring hvKp isolates, which may be used as a prospective alternative control strategy to fight this ominous bacterium.Keywords: Klebsiella pneumoniae, bacteriophages, colistin, MDR, virulence