ISOLATION AND CHARACTERIZATION OF A NOVEL BACTERIOPHAGE TO CONTROL MULTIDRUG-RESISTANT ACINETOBACTER BAUMANNII
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Keywords
Clinical, bacteriophage, host range, resistance, applications
Abstract
In order to tackle the urgent problem of multidrug-resistant (MDR) Acinetobacter baumannii infections, this work describes Phage-1, a lytic bacteriophage that has strong anti-bacterial properties. Patients in the intensive care unit with suspicision of septicemia had bacterial samples taken, and antibiotic susceptibility testing was carried out in accordance with CLSI/USA guidelines. Automated VITEK 2 system was used to assess the antibiotic susceptibility of 60 isolates of A. baumannii. Phage-1 was isolated from sewage system of a tertiary care hospital in Faisalabad using conventional screening techniques. Centrifugation, binary layer agar identification, and many rounds of isolation were used in the phage extraction and purification procedure. The Phenol Chloroform technique was used to extract DNA from Phage-1. Using a one-step growth curve analysis, the life cycle of Phage-1 was deciphered. This revealed a latent time of 30 minutes, a lysis period of 60 minutes, and a burst size of 8.6 PFU/mL. Phage-1 was shown to be resilient in thermal and pH stability testing, exhibiting substantial activity at 20-40°C and 6-8 pH as well as vulnerability to higher temperatures and severe pH values. Sixty distinct strains of MDR A. baumannii were subjected to host range studies, which demonstrated Phage-1's capacity to lyse local strains within a given geographic area. Minimum Inhibotory Concentration (MIC) was assessed using VITEK 2 system, which showed that the mostly isolates were resistant to cephalosprins, carbapenems and aminoglycosides. All strains are sensitive to Polymyxin B. This study adds important new information on how Phage-1 may be used as a therapeutic agent to treat MDR A. baumannii infections. The results show its lytic efficiency, stability, and geographic uniqueness, emphasizing the need of taking regional factors into account when developing phage-based therapies to fight bacterial illnesses resistant to antibiotics.
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Asma Yaqoob
Consultant Pathologist, THQ Hospital Shahkot, Pakistan
Aroosh Shabbir
Assistant professor, Institute of Molecular Biology and Biotechnology, University of Lahore, Punjab
Muhammad Aamir Aslam
Assistant professor, Institute of Microbiology, University of Agriculture Faisalabad.
Namra yunus
Assistant professor, Fatima Memorial Medical and Dental College, Lahore.