Pulsed-Field Gel Electrophoresis Is More Efficient than Ribotyping and Random Amplified Polymorphic DNA Analysis in Discrimination ofPasteurella haemolyticaStrains

Abstract

One hundred thirty-three strains of Pasteurella haemolytica of both biotypes (90 and 43 strains of biotypes A and T, respectively) and almost all the serotypes were subjected to ribotyping, random amplified polymorphic DNA (RAPD) analysis, and pulsed-field gel electrophoresis (PFGE) analysis for epidemiological purposes. A total of 15 patterns recorded as ribotypes HA to HO were found for the P. haemolytica biotype A strains, with ribotypes HA, HC, and HD being encountered most often (66 strains [74%]); and 20 ribotypes, designated HA′ to HT′, that were clearly distinct from those observed for biotype A strains were observed for strains of biotype T. RAPD analysis generated a total of 44 (designated Rp1 to Rp44) and 15 (designated Rp1′ to Rp 15′) unique RAPD patterns for biogroup A and biogroup T, respectively. Analysis of the data indicated that a given combined ribotype-RAPD pattern could be observed for biotype A strains of different serotypes, whatever the zoological or geographic origin, whereas this was not the case for biotype T strains. PFGE appeared to be more efficient in strain discrimination since selected strains from various zoological or geographical origins harboring the same ribotype-RAPD group were further separated into unique entities.

Characterization of United Kingdom Isolates ofCorynebacterium pseudotuberculosisUsing Pulsed-Field Gel Electrophoresis

“…The reproducibility of the PFGE method was confirmed by repeated testing of a panel of five isolates. 2,3,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41,42,43,44 Inserts prepared from the same batch of plugs, when run on eight different gels, yielded identical banding patterns. The in vitro stability of the pulsotypes was confirmed in that the banding patterns before and after 20 passages were identical.…”

Serotypes A1 and A2 ofMannheimia haemolyticaare susceptible to genotypic, capsular and phenotypic variations in contrast to T3 and T4 serotypes ofBibersteinia (Pasteurella) trehalosi

“…In some cases, strains belonging to the same serotype have been found in various animal hosts, i.e. serotype A1 in bovine, ovine and caprine hosts (Chaslus-Dancla et al, 1996;Fodor et al, 1999;Kodjo et al, 1999) as well as T3 in ovine, caprine and wild ruminant hosts such as chamois (Rupicapra rupicapra) (Foreyt & Jessup, 1982;Snipes et al, 1992). This suggests a possible zoological interspecies distribution of any given strain.…”

“…To assess the transmission of a Pasteurella strain from one animal host to another, a convenient epidemiological tool able to take into account all possible variations within a given strain is necessary. The authors have previously demonstrated the increased accuracy of pulsed-field gel electrophoresis (PFGE) analysis compared with ribotyping and random amplified polymorphic DNA in M. haemolytica strain discrimination (Kodjo et al, 1999). This study is aimed at refining this molecular tool and simulating in vitro the plasticity of certain strains of M. heamolytica and B. trehalosi through successive subcultures in order to address objective criteria for use in epidemiological investigations.…”

Use of REP‐PCR and 16S rRNA gene sequencing for comparison of Mannheimia haemolytica isolates obtained from fatal cases of bovine respiratory disease in the USA and Australia

“…A variety of techniques have been used to characterise M. haemolytica , including PFGE, REP‐PCR and MLST . These tools have found wide ranges in diversity among M. haemolytica isolates.…”

“…true differences in diversity influenced by anatomical, animal or geographical origins) or artefacts of the tools. Although PFGE is more discriminatory and can identify greater diversity within a given group of isolates, the relevance and utility of this increased discrimination is unclear. Thus, it would appear that no approach has been established as the best epidemiological tool.…”