The comparison and characterisation of glycolytic mycoplasmas isolated from the respiratory tract of sheep

Abstract

PLATES 111 AND IV THE original isolations of glycolytic mycoplasmas in Scotland were made by Mackay, Nisbet and Foggie (1963) from cases of sheep pulmonary adenomatosis (SPA). Identical (" Type A ") mycoplasmas were later isolated from pneumonic but not from normal lungs of sheep, and from the upper respiratory tract of apparently healthy sheep (Mackay, cited by Cottew and Leach, 1969). Krauss and Wandera (1 970) compared mycoplasmas recovered from cases of SPA in Kenya with other mycoplasrnas, including the Scottish MS strain originally isolated by Mackay from a case of SPA. The Scottish strain was placed in one of three serogroups.Mycoplasmas recovered from lambs and sheep in Queensland, Australia, by St George et al. (1971) have been characterised by Carmichael et al. (1972), and the name MycopZasma ovipneumoniae proposed. Glycolytic mycoplasmas isolated from the respiratory tract of sheep in Victoria, Australia (Cottew, 1971; Furlong and Cottew, 1973), and in New Zealand (Clarke, Brown and Alley, 1974) were identical to the Queensland strains of M. ovipneumoniae.The purpose of this investigation was to compare the biochemical and serological reactions of glycolytic mycoplasmas isolated from SPA cases, pneumonic sheep and apparently healthy sheep in Scotland with a Queensland strain of M. ovipneumoniae. MATERIALS AND METHODSStrains examined. These are listed in table I. All strains isolated in this laboratory were cloned at least four times by serial transfer of single colonies from agar plates to broth. The possibility that the strains were L-forms was eliminated by five consecutive subcultures on solid medium (OA) from which bacterial inhibitors had been omitted. No reversion to bacterial forms occurred.Isolation media. Broth medium (OB) comprised (v/v) 20 % of Brain-Heart Infusion (Oxoid Ltd., London), 60 % of Medium 199 (Burroughs Wellcome, Beckenham, Kent), 10% of inactivated swine serum and 10% of fresh yeast extract (Marmion, 1967), supplemented with ampicillin, 1 mg per ml (" Penbritin ", Beecham Veterinary Products, Crawley, Sussex) thallium acetate 0.025 % (w/v) and phenol red 0.006 % (w/v). The medium was adjusted to pH 7.6-7-8 with M NaOH. Solid medium (OA) differed in containing 15% (v/v) (w/v). The Agarose was autoclaved with the Brain-Heart Infusion and the other constituents were added when the suspension had cooled to 56°C.Biochemical test media. The basic media, EA and EB, were based on OA and OB respectively, but with Eagle's Minimal Essential Medium (MEM) substituted for Medium 199 and phenol red omitted unless specifically required. All strains were subcultured twice in EB before being tested. Paired broth cultures were incubated aerobically and, by being overlaid with sterile paraffin, anaerobically. Paired agar plates were incubated, one in a candle-jar and the other in an atmosphere of 95% N2 and 5 % COZ, as described by Aluotto et al. (1970).For all biochemical tests, controls included test media inoculated with sterile broth or agar blocks.Rabbit broth (RB). For the production o...

Comparison of antigens of pneumonia-associated mycoplasma species by gel diffusion

“…This present work confirms that this relationship is two way. Similarly, although Jones et al (7) was able to demonstrate a cross-reaction between M. ovipneumoniae antiserum and M. hyorhinis antigen, only the reverse cross was shown in this present work.…”

“…In all of the Mycoplasma species investigated, the species-specific antigens detected by MI or GI tests have been shown to be membrane associated (6,8,13,15). No cross-reactions have been shown by these tests with the six species described here (5,7,11). MI and GI tests carried out with the antisera raised for this study showed only homologous reactions by M. bovoculi and M. hyorhinis antisera to their respective antigens (unpublished data).…”

NOD2/c-Jun NH 2 -Terminal Kinase Triggers Mycoplasma ovipneumoniae-Induced Macrophage Autophagy

“… M. ovipneumoniae Queensland strain Y98 ( M. ovipneumoniae Y98) was purchased from the China Institute of Veterinary Drug Control (Beijing, China). M. ovipneumoniae was cultured in a mycoplasma broth containing mycoplasma broth base CM403, supplementary reagent SR59 (Oxoid, Hampshire, UK), 0.5% glucose, and 0.002% phenol red at 37°C with 5% CO 2 ( 48 ).…”

Mycoplasma ovipneumoniae induces sheep airway epithelial cell apoptosis through an ERK signalling-mediated mitochondria pathway

“…The M. ovipneumoniae Queensland Strain Y98 [ 58 ] was obtained from the China Institute of Veterinary Drug Control (Beijing, China). As previously described [ 58 ], the mycoplasma bacterium was cultured and propagated in a mycoplasma broth containing mycoplasma broth base CM403, supplement-G SR59 (OXOID, Hampshire, UK), 0.5 % glucose, and 0.002 % phenol red at 37 °C in 1 % CO 2 . The titre of M. ovipneumoniae culture was determined based on the metabolic activity of the bacterial cells in the medium, presented as a colour change unit (CCU)/mL [ 59 ].…”