Characterisation of Histophilus ovis and related organisms by restriction endonuclease analysis

Abstract

The banding profiles generated by Bam H1 restriction endonuclease cleavage of bacterial DNA from clinical and reference isolates of Histophilus ovls, Haemophilus somnus and related bacteria were compared. H. ovis, H. somnus and Haemophilus agni isolates were found to have distinct similarities in banding profiles characterised by 10 common bands between 2.0 and 9.6 kilobases (kb). The close taxonomic relationship of these isolates was reinforced by these findings. The reference isolates examined In this study -Actinobacillus lignieresii, Actinobaclllus semlnis, H. agni, H. somnus, H. ovis, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus parahaemolyficuscould be distinguished from each other on the basis of their characteristic banding profiles.Aciinobacillus sp were obsewed to have more bands between 2 and 23 kb compared with the H. ovis and Haemophilus sp isolates studied.Analysis of isolates from an experimental infection trial illustrated the potential of restriction endonuclease analysis in molecular epidemiological applications. It was possible to demonstrate by this means that the post-challenge isolates had identical banding profiles to the challenge (or infecting) isolate which had a distinctly different banding profile from that of pre-challenge H. ovis isolates. Furthermore, restriction endonuclease analysis of H. ovis isolates obtained from follow-up investigations of a recurrent problem of epididymitis in unmated rams, indicated that the H. ovis isolates implicated in epididymitis, were present as a single strain in a number of sheep over a period of time. This suggested that the mechanism of transmission was by perinatal preputial contamination. Aust Vet J 6 3 389-393 Results Barn HI Profiles of H. ovis and Closely Related BacteriaExamples o f the banding profiles obtained from Bam HI digests o f DNA from H. ovis (Australian) and H . somnus (North American) isolates are shown in Figure 1. The profiles o f the H . somnus isolates (lanes 2 and 3) are different from $ Seakem-LE Agarose, SUMMARY Wet chemical tests have deficiencies when applied to mixtures containing silica, which are common in the uroliths of some domestic animals. Consequently, the applicability of an infrared spectroscopic method was tested on 104 uroliths obtained from cattle, sheep, goats, horses, pigs, dogs, a chicken and a rabbit during diagnostic investigations. The following components were satisfactorily identified: silica, calcium oxalate, calcium carbonate, calcium phosphate, magnesium ammonium phosphate, magnesium phosphate and urates. The Infrared characteristics of these compounds and their mixtures are described. Aust Vet J 63: 393-396

Acquisition of haemoglobin-bound iron by Histophilus somni

“…For example, Stephens et al (1983) showed that H. ovis, H. somnus and H. agni are very c10sely related biochemicaIly and these findings were later supported by DNA hybridization studies (Walker et al, 1985;PiechuUa et al, 1986) and restriction endonuc1ease analyses (McGillivery et al, 1986;Kirkham et al, 1989). Interestingly, H. ovis and H. somnus can aIso be differentiated by restriction enzyme analyses (McGillivery et al, 1986;Kirkham et al, 1989) and by means ofpolymerase chain reaction (peR) ribotyping (Appuhamy et al, 1998). In 2003, Angen and co-workers reported the grouping of aU three species based on the nuc1eotide sequences of 16S rRNA and the genes (ropB) that encode the ~ subunit of RNA polymerase.…”

Characterization of immunodominant surface antigens of Haemophilus somnus

“…This result is consistent with the antigenic (4,32,38) and biochemical similarities of H. somnus and H. agni (2,38) which have been shown by others. More recently, H. somnus, H. agni, and Histophilus ovis were shoWn to be very similar by restriction endonuclease analysis (31). Historically, isolates were called H. agni or Histophilus ovis if obtained from sheep and H. somnus if obtained from cattle (2,38); therefore, the cross-reactivity between H. somnus and H. agni should not lead to confusion in diagnostic assays.…”

Genomic fingerprinting of "Haemophilus somnus" isolates by using a random-amplified polymorphic DNA assay

“…Multilocus enzyme electrophoresis has not been undertaken with these organisms. Plasmid typing and restriction endonuclease fingerprinting have been used to differentiate among a small number of isolates of "H. somnus," "H. agni," and "H. ovis" (16,21).…”