title: Isolation and Characterization of Lytic Properties of Bacteriophages Specific for M. haemolytica Strains.
abstract: OBJECTIVE
  The objective of this study was isolation and morphological characterization of temperate bacteriophages obtained from M. haemolytica strains and evaluation of their lytic properties in vitro against M. haemolytica isolated from the respiratory tract of calves.
  
  
  METHODS
  The material for the study consisted of the reference strain M. haemolytica serotype 1 (ATCC®) BAA-410™, reference serotypes A1, A2, A5, A6, A7, A9 and A11, and wild-type isolates of M. haemolytica. Bacteriophages were induced from an overnight bacterial starter culture of all examined M. haemolytica strains treated with mitomycin C. The lytic properties and host ranges were determined by plaque assays. The morphology of the bacteriophages was examined in negative-stained smears with 5% uranyl acetate solution using a transmission electron microscope. The genetic analysis of the bacteriophages was followed by restriction analysis of bacteriophage DNA. This was followed by analysis of genetic material by polymerase chain reaction (PCR).
  
  
  RESULTS
  Eight bacteriophages were obtained, like typical of the families Myoviridae, Siphoviridae and Podoviridae. Most of the bacteriophages exhibited lytic properties against the M. haemolytica strains. Restriction analysis revealed similarities to the P2-like phage obtained from the strain M. haemolytica BAA-410. The most similar profiles were observed in the case of bacteriophages φA1 and φA5. All of the bacteriophages obtained were characterized by the presence of additional fragments in the restriction profiles with respect to the P2-like reference phage. In the analysis of PCR products for the P2-like reference phage phi-MhaA1-PHL101 (DQ426904) and the phages of the M. haemolytica serotypes, a 734-bp phage PCR product was obtained. The primers were programmed in Primer-Blast software using the structure of the sequence DQ426904 of reference phage PHL101.
  
  
  CONCLUSIONS
  The results obtained indicate the need for further research aimed at isolating and characterizing bacteriophages, including sequence analysis of selected fragments. Moreover, standardization of methods for obtaining them in order to eliminate M. haemolytica bacteria involved in the etiopathogenesis of BRDC is essential.
authors.affiliation: Sub-department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033, Lublin, Poland.
authors.forename: Renata
authors.initials: R
authors.lastname: Urban-Chmiel
authors.affiliation: Sub-department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033, Lublin, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Wernicki
authors.affiliation: Sub-department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033, Lublin, Poland.
authors.forename: Diana
authors.initials: D
authors.lastname: Stęgierska
authors.affiliation: Sub-department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033, Lublin, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Dec
authors.affiliation: Sub-department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033, Lublin, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Dudzic
authors.affiliation: Sub-department of Veterinary Prevention and Avian Diseases, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, 20-033, Lublin, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Puchalski
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D002417:Cattle; D002457:Cell Extracts / Q000302:isolation & purification*; D058507:Host Specificity; D016978:Mannheimia haemolytica / Q000302:isolation & purification* / Q000821:virology*; D012137:Respiratory System / Q000382:microbiology; D000095983:Bacterial Lysates
chemical_list: D002457:Cell Extracts; C030259:Broncho-Vaxom; D000095983:Bacterial Lysates