Background: The use ofbacterial viruses has become of major interest in treatment of multi drugpathogens such as klebsiella pneumoniae. We studied thesusceptibility of klebsiella pneumoniae to a mixture of two lyticbacteriophages isolated from a hospital waste water treatment plant.Methods: Phage isolation was carried out by takingsamples from the outlets of three hospital wastewater treatment plants.
Thesamples were filtered using 0.45?m filters after centrifugation at 6,000rpm,10min. An overnight culture of Klebsiella pneumoniae ATCC 10031 (volume5ml) was mixed with1ml Volume of the filtered water samples and incubated for 24h at 370Cbefore centrifugation at 6000Rpm andfiltering through a 0.22?m membrane filter. Titration of the lytic phages inthe filtrates was examined by the double layer method using 100µl ofthe diluted virus with 400µl of the ATCC strain (0.
5 McFarland) in 5ml of 0.75%molten soft agar before layering the mixture on nutrient agar plates. Theplates were incubated at 37°C overnight and the resulting plaque forming units(PFU) were then counted. The phage mixture was then centrifuged at 6000rpm andthe pellet was washed using ammonium acetate (0.1 M, pH 7.0). A portion of theresuspended sediment was deposited on formvar carbon coated grid Cu Mesh 300,stained with 2% uranyl acetate and examined in EM10C (Zeiss, Germany) transmissionelectron microscope at 100kV.
Results: Two lytic bacteriophages were isolated whichformed 1 and 2 mm plaques when plated against the klebsiella pneumoniaehost strain. The TEM results showed that the isolated phases resembled thetailed bacteriophages of Siphoviridae and Myoviridae families. Conclusions: The bacteriophages isolated in this researchshowed specificity towards klebsiella pneumoniae ATCC hoststrain. Further research is underway to examine their potential use againstmulti drug resistant klebsiella pneumoniae.