Isolation of Klebsiella pneumoniae phage from sewage and analyses its biological and genomic characteristics
To explore the potential of phages as a novel therapeutic approach for Klebsiella pneumoniae infec-tions,this study employed the double-layer plate method to isolate and purify three strains of lytic Klebsiella pneumoniae phages from the sewage surrounding the hospital.The morphological structure of the phages was observed using transmission electron microscopy,and further analysis was conducted on their biological char-acteristics,including lytic spectrum,optimal infection multiplicity,growth curve,and acid-base tolerance.Additionally,sequencing technology was utilized to obtain and annotate the complete genomic information of the phages.Experimental results indicate that KP-ZS3 is a Myoviridae phage,KP-ZS4 is a Podoviridae phage,and KP-ZS6 is a Siphoviridae phage.KP-ZS4 has a slightly longer latent period of infection,KP-ZS3 has a stronger acid-base tolerance,and KP-ZS6 has the strongest lytic capability and the broadest lytic spec-trum among the three phages.The capsid protein components of all three phages are within the range of 33 kD to 55 kD.Moreover,genomic analysis revealed that the genomes of KP-ZS3 and KP-ZS6 are both larger than 100 kb,with the genome of phage KP-ZS4 being relatively smaller(40608 bp).Comparative genomic analysis shows that phage KP-ZS3 is highly similar to phiEap-3 and Kp15;KP-ZS6 is highly similar to 0507KN21 and vB_EcoM-KWBSE43-6.KP-ZS4 has a relatively lower similarity to Tyrion and TL-2011b,with sequence identities of 76.77%and 76.89%,respectively.These results suggest that these three phages possess unique biological characteristics and genomic features,demonstrating different potentials and advan-tages in the treatment of Klebsiella pneumoniae infections.In particular,KP-ZS6,with its strong lytic ability and broad lytic spectrum,and KP-ZS3,with its strong acid-base tolerance,may serve as alternative re-sources for future antibacterial treatments.These findings provide a new informational basis for the develop-ment of novel antibacterial strategies.
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