Isolation and characterization of novel Klebsiella phages from Benin and their antibiofilm activities on multidrug resistant and hypervirulent Klebsiella pneumoniae strains

Hypervirulent Klebsiella pneumoniae represents a growing clinical threat, particularly where multidrug resistance limits treatment options. This study aims to characterize Klebsiella phages isolated in Benin and evaluate their efficacy in reducing bacterial cell growth, biofilm inhibition and biofilm disruption. To achieve this objective, we isolated Klebsiella phages by enriching samples of hospital effluent, pig droppings, and stool, using clinical multidrug resistant and hypervirulent K. pneumoniae strains as hosts. These phages were characterized using transmission electron microscopy, whole-genome sequencing, and biological assays. Then, we evaluated their antibiofilm activity and stability under harsh conditions (pH and temperature) in vitro . So, we report the isolation and characterization of three novel lytic phages from Benin (Kp1Bj_HH11_M23, Kp2Bj_LN294_M23, and Kp10Bj_LN54_14). These Myoviruses, belonging to the genera Marfavirus and Jiaodavirus , display broad host range activity against multidrug resistant and hypervirulent K. pneumoniae strains with various capsular serotypes including KL114, KL53, KL8, KL62, KL64, KL124, KL102, KL57, KL39, KL48, KL17, KL20, KL2, KL30, KL51, KL105, KL146, KL28, KL122, KL38, KL3, and KL163. Genomic analysis confirmed the absence of virulence and antimicrobial resistance genes. The phages exhibit rapid adsorption, short latency periods (5–10 min), and high burst sizes (119–2208 PFU/cell). All three phages have significantly biofilm disruption effect in vitro (Kp1Bj_HH11_M23, biofilm disruption rate of 98.46 %, p -value = 0.0170; Kp2Bj_LN294_M23, biofilm disruption rate of 65.56%, p -value < 0.0001; Kp10Bj_LN54_14, biofilm disruption rate of 99.96%, p -value = 0.003). Their stability across a wide range of temperatures and pH further supports their potential for therapeutic development. Together, these data highlight the value of locally sourced phages as candidates for tackling region-specific antimicrobial resistance challenges.