Production of Anti-Mycoplasma (PPLO) Antibodies in Rabbits.

Morton, Harry E.; Roberts, Ruth J.

doi:10.3181/00379727-125-32140

Cited by 61 publications

(19 citation statements)

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“…All strains in Antiserum production. New Zealand rabbits were vaccinated by the method of Morton, and Roberts (1967), 3 ml of antigen in adjuvant being given by injection into the muscle, dermis and footpad, followed 3 weeks later by 2 ml intramuscularly. The growth-inhibiting titres obtained by this technique alone were unsatisfactory however, and two supplementary i.v.…”

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

Jones

Foggie

Mould

et al. 1976

Journal of Medical Microbiology

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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...

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

Jones

Foggie

Mould

et al. 1976

Journal of Medical Microbiology

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“…The use of adjuvants has long been advocated for the production of high-titred mycoplasma antisera (Morton & Roberts, 1967) but some species of mycoplasma consistently fail to produce certain types of anti-mycoplasma antibodies in rabbits (Carroll, Rollins & Jasper, 1976) even when an adjuvant is used. The results presented reveal that for M. bovis the use of an adjuvant is desirable for production of specific precipitins in the GP and GD tests, but it was also shown that a number of anti-medium precipitins were produced which cross-reacted with M. bovigenitalium antigen.…”

Section: Discussionmentioning

confidence: 99%

Studies on the effect of growth medium composition on the antigenicity ofMycoplasma bovis

Thorns

Boughton

1980

J. Hyg.

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SUMMARYSerum proteins adsorbed from the culture medium were detected in Mycoplasma bovis antigens, the number and type of proteins depending on the serum used in the medium. The c-globulins cross-reacted with a-globulins from different types of sera but the y-globulins did not. The removal of non-specific medium antibodies by absorption showed that they affected the gel diffusion and growth precipitation tests, producing cross-reactions between M. bovis and M. bovigenitalium, but that the complement fixation, tube agglutination, and growth inhibition tests were not similarly affected. The presence of serum proteins in the antigens changed their specific reactivity in all the tests. The production of antibodies to serum proteins was increased by the use of an adjuvant, but it appeared that production of specific antibodies to the mycoplasmas was not.

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“…Antisera against Anaeroplasma strains 6-1, 7LA, JR, and 161 were prepared in rabbits. The inoculation procedure was similar to that described by Morton and Roberts (8). The sites of inoculation, amounts of inoculum, and intervals between inoculations are indicated in Table 2.…”

Section: Methodsmentioning

confidence: 99%