Characterization of E coli phage isolated from sewage

Abstract

Bacteriophage are viruses that infect bacterial cells. as with all viruses, phage are nonliving agents and thus require the use of the host‟s metabolic processes to replicate itself. in this study, the phage of interest are those that infect and lyses E. colt host cells. when phage are released from the ruptured host, distinct zones of clearing (plaques) form. the original E. colt host cells for this experiment came from a sample of raw sewage. in order to obtain the bacteriophage, a procedure of enrichment, isolation, dilution and seeding was followed, the presence of distinct plaques indicated that lytic bacteriophage had been successfully amplified, separated and grown.This study included determination of phage titre, latent period , rise period and the burst size of the phage and effect some of factor on phage titre such as (temperature, ether and chloroform) .for determination ofhage titre used series of dilutions(10-1, 10-2, 10-3, 10-4, 10-4, 10-6, 10-7, 10-8, 10-9) the dilution factor gave the best countable number of plaques is(103). this dilution factor was then used for all other experiments, the latent period , rise period and the burst size of the phage are determined by countable number of plaques and phage titre(titer: plaque-forming unit(p.f.u) during 10,20,30,40,50, and 60 minutes . it was (4.7x105 „ 5.3x105 and 6.0x105)during 1O,20and30minutes respectively in the latent period ,but it was (8.5x105 8.9x10‟ 9.3x105)during 40,50,and 60 minutes respectively in the rise period .then the burst size of the phage is counted by the ratio of the phage titer after rise period to that during the latent period it was(1.67).This study also included effect of temperature on phage titre the statistical analysis was significantly increase P<0.05 in phage titre at the temperature37 C° comparing with phage titre at the temperature 50 C° and phage titre at the temperature 65 C°. effects of ether and chloroform on number of plaques and phage titre during 5,10,15 ,20,25 ,30,35 and 40 minutes it was(0.7x105 , 0.3x105 , 0 , 0 , 0 , 0, 0 and 0) respectively in ether sensitivity, but the phage titre in chloroform sensitivity was completely inactivated by chloroform treatment, the statistical analysis (freedom degree ( 2,21 ) and F value=52.60 was high] significant increase (P<0.05) in phage titre in normal saline comparing with phage titre in ether and chloroform sensitivity