Date Published: June 19, 2018
Publisher: Springer Berlin Heidelberg
Author(s): Renata Urban-Chmiel, Andrzej Wernicki, Jacek Wawrzykowski, Andrzej Puchalski, Anna Nowaczek, Marta Dec, Diana Stęgierska, Mohammed Mijbas Mohammed Alomari.
The aim of study was to isolate, characterize and analyse the protein profiles of Myoviridae-like bacteriophages obtained from M. haemolytica using MALDI TOF mass spectrometry. The material consisted of the M. haemolytica reference strain ATCC® BAA410, reference serotypes A1, A2, A5, A6, A7, A9, and A11, and wild-type isolates of serotype A1. Bacteriophage morphology was examined with a transmission electron microscope. The proteins were separated in SDS-PAGE and two-dimensional electrophoresis and characterized by MALDI-TOF. Among the phages obtained, seven were specific for strains A1, A2, A5, A6, A7 and 25, and PHL-1 was specific for the BAA410 strain. The protein profiles for the phages were very similar to one another, but differed from the reference phage in that they lacked protein fractions with molecular weights of 22.9, 56.3 and 73.1 kDa. 2D electrophoresis revealed significant differences in the size of proteins and their localization in the pH gradient. The most similar profiles were observed in phages specific for strains BAA-410 and A6. In all profiles two main spots were observed in the molecular weight range from 44 to 70 kDa at pH < 4. The results indicate that 2D electrophoresis is a very useful tool for characterization of phage protein profiles. An important objective was to determine the molecular differences between morphologically similar phages belonging to one family and to find similarities to phages specific for other pathogens. The study also assessed the suitability of the methods used to characterize phages.
All types of bacteria exhibit susceptibility to bacteriophages specific to them. Numerous studies have shown that bacteriophages can be isolated from a variety of environments in which bacteria are present. Examples include natural water bodies, wastewater, soil, plants and animals (Kutateladze and Adamia 2010; Leverentz et al. 2004).
Analysis of the phage isolates induced from M. haemolytica showed slight variation in protein profiles in SDS-PAGE electrophoresis and significant differences in protein profiles in 2D electrophoresis. This is interesting in that the bacteriophages obtained were all identified as belonging to the family Myoviridae on the basis of morphological analysis under an electron microscope.