Isolation and characterization of Klebsiella pneumoniae specific bacteriophages from sewage samples

Abstract

Five bacteriophages (Kpn5, Kpn12, Kpn13, Kpn17 and Kpn22), each having specificity against Klebsiella pneumoniae strain B5055, were isolated from sewage samples and characterized in terms of growth characteristics, genetic material, morphology and structural proteins. Adsorption rate as well as single step growth curve experiments showed variation among phages. Restriction enzyme digestion of DNA confirmed the presence of double stranded DNA as well as the heterogeneous nature of genetic material. RAPD-PCR was performed to further distinguish these closely related phages. Their genome fingerprint confirmed their diversity. Transmission electron microscopy, on the other hand, showed their morphological similarity; they were assigned to family Podoviridae, order Caudovirales on the basis of their head and tail morphology. Structural proteins resolved on SDS-PAGE showed the presence of similar major outer membrane proteins. The bacteriophages, belonging to Podoviridae family with short stumpy tails, were found to be nontoxic to mice. They showed maximum count in various organs at 6 h post inoculation, which persisted till 36 h. These phages thus have the potential to be used for phage therapy.

Isolation and characterization of a lytic bacteriophage φKp-lyy15 of Klebsiella pneumoniae

“…This study describes a newly isolated bacteriophage φKp-lyy15 specifi cally infecting K. pneumoniae isolates and its biological features. The Klebsiella bacteriophages described in publications belong to Podoviridae, Myoviridae and Siphoviridae families (Ackermann, 2001;Kumari et al, 2010). Our results showed that phage φKp-lyy15 has an icosahedral head 50 nm in diameter and a filamentous tail 120 nm in length.…”

Isolation and characterisation of KP34—a novel φKMV-like bacteriophage for Klebsiella pneumoniae

“…There is a need to look for novel phages with lytic potential to multidrug resistance K. pneumoniae isolates. Recently isolated Klebsiella phages belonged to Podoviridae family (Kumari et al 2009; 2010a; b; Verma et al 2009) or pseudo-T-even family (Wu et al 2007). During our team’s experiments, five different viruses from the Podoviridae , Siphoviridae and Myoviridae were characterised regarding their biology and genomics (data not shown) and genomes of three phages have already been deposited in GenBank.…”

“…Similar results were also obtained with the T7-like Pseudomonas phage φIBB-PF7A (Sillankorva et al 2008). Other investigators analysing structural proteins of K. pneumoniae T7-like phages by SDS-PAGE technique obtained wide range of band numbers, from 12 (29 to 120 kDa) with K. pneumoniae phage B5055 (Verma et al 2009) to two to five bands representing proteins from 20 to 55 kDa in five phages (Kumari et al 2010b). This would suggest that considerable variation occurs in this group of bacteriophages with respect to protein number and size.…”

“…Data concerning newly isolated Klebsiella phages focus on biological characteristics or biodistribution and therapeutic efficacy in animal models (Bogovazova et al 1991; Kumari et al 2009; 2010a, b; Verma et al 2009). None of the above described Klebsiella phages was tested regarding genome organisation or sequence similarity to already known viruses.…”

The influence of external factors on bacteriophages—review

“…They can be found in all environments where bacteria grow: in the Sahara, hot springs, the North Sea, and polar inland waters (Prigent et al 2005; Lin et al 2010; Breitbart et al 2004; Wichels et al 1998; Säwström et al 2008). Phages are detected in ground and surface water, soil, food (e.g., sauerkraut, wine), sewage, and sludge (Lucena et al 2006; Yoon et al 2002; Davis et al 1985; Kumari et al 2010; Tartera and Jofre 1987). They have also been isolated from humans and animals, for example from feces, urine, saliva, spit, rumen, and serum (Gantzer et al 2002; Caroli et al 1980; Bachrach et al 2003; Nigutová et al 2008; Keller and Traub 1974).…”