Presence of a dialysable fraction in normal bovine whey capable of killing several species of bovine mycoplasmas

International Journal of Systematic Bacteriology

Wyld

Leach³

1978

Self Cite

Eight similar Mycoplasma isolates were obtained from rectal, intestinal, and vaginal specimens from pigs. Two indistinguishable clones of one isolate (Mayfield) were examined in detail. Although closely resembling the cattle organism Mycoplasma alvi in cultural and biochemical properties, the Mayfield clones differed from this species in morphological appearance and deoxyribonucleic acid base composition. The clones were serologically distinct from M. alvi and 58 currently recognized Mycoplasma species and subspecies, including all of those having similar biochemical properties or the same host. The Mayfield strain clone B (NCTC 10170) is designated the type strain of a new species, M. sualvi, which includes five of the seven other Mycoplasma isolates from pigs. The remaining two isolates appear to be serologically dissimilar from the Mayfield strain.Three strains of Mycoplasma were isolated from porcine rectal swabs, and the partial characterization of one of them (the Mayfield strain) suggested that they might represent a new species (1). This paper presents a detailed characterization of two clones of the Mayfield strain and proposes their classification, together with five other similar isolates from pigs, as a new Mycoplasma species. MATERIALS AND METHODSMycoplasma strains studied. The Mayfield strain was isolated from a porcine rectal swab as previously described (I). Before testing, it was "fdtercloned" three times (2), and two separate clones (A and B) were examined for characterization purposes.Strain H770 was isolated from the vagina, strain 2495 was isolated from the colon, strains PF1 and PF3 were isolated from the rectum, and strains 1828,2284, and 2685 were isolated from the small intestine of pigs.The other Mycoplasma species and antisera used in comparative studies are listed in Table 1.Media. Except where indicated, U3/7.0 broth and U3 agar plates (2,3) were used. The main ingredients of these media are fetal calf serum, Hartley digest broth, and yeast extract. Methods. Except where otherwise indicated in thetext, all materials and methods were as described for the isolation and characterization of Mycoplasma alvi (2). RESULTSCultural characteristics and reversion studies. The Mayfield clones had cultural characteristics similar to those of the bovine isolate M. alvi (2). When subcultured in U3/7.0 broth, the Mayfield clones produced both opacity and an acid pH color change (1.5 pH units) after 3 to 4 days of incubation at 37°C. Unlike M. alvi, the porcine strains produced an acid color change (0.8 pH unit) in glucose serum (GS) broth (pH 7.8) (3). However, acidification in the U3/7.0 broth occurred more rapidly than in GS broth. In arginine broth (2), the porcine strains produced an initial acid color change (1.5 pH units) after 3 to 6 days of incubation, followed after 2 to 3 weeks by an alkaline reaction manifested by a return to the original pH of the medium (pH 7.0). The MayField clones grew less rapidly in U3/7.0 broth at 30°C than at 37"C, and no color change was observed after 70 days of...

Section: Resultsmentioning

confidence: 99%

Mycoplasma sualvi, a New Species from the Intestinal and Urogenital Tracts of Pigs

International Journal of Systematic Bacteriology

Wyld

Leach³

1978

Self Cite

“…This may reflect the differing ability of mice to deal with inoculated organisms in the early stages of infection. It is possible that humoral mycoplasmacidal factors, similar to those found in bovine whey (Brownlie, Howard and Gourlay, 1974;Howard et al, 1975) might be involved in host defence at this stage of the infection, particularly as neutrophils were found in the glands of only one of five BSVS mice 24 h after inoculation with U. urealyticum T960.…”

Section: Discussionmentioning

confidence: 99%

Production of Mastitis in Mice With Human and Bovine Ureaplasmas (T-Mycoplasmas)

Howard

Anderson

Journal of Medical Microbiology

et al. 1975

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D. TAYLOR-ROBINSON?PLATE XLV UREAPLASMAS (T-mycoplasmas) (Shepard et al., 1974) of bovine origin cause experimental mastitis in cows (Gourlay, Howard and Brownlie, 1972;Howard, Gourlay and Brownlie, 1973). In contrast, none of six ureaplasma strains of human origin was capable of producing mastitis in cattle although two of these strains produced mastitis in goats . Cattle and goats, however, are not readily obtainable for experimental work and the use of small laboratory animals would greatly facilitate studies on ureaplasma pathogenesis and immunology. Mastitis has been produced in mice by intramammary inoculation of bacteria obtained from clinical cases of bovine mastitis (Chandler, 1970), and the purpose of this investigation was to determine whether it was possible to produce mastitis in mice with human and bovine ureaplasmas. MATERIALS AND METHODSUreaplasma strains. Strains derived from the human genital tract, namely nos. CD408, M126/68 and CD573, have been described previously . Strain CD591 (Johnson) was isolated from the human oropharynx (Taylor-Robinson and Purcell, 1966) and strain T960 is the type strain of Ureaplasma urealyticum (Shepard et al., 1974). Strains A417, Vic9, Bu2 and BlOl were isolated from cattle (Howard et al., 1973). Ureaplasmas were grown at 37°C in either U3 broth and the corresponding solid medium (Howard et al., 1975) or a PPLO medium described by Taylor-Robinson, Addey and Goodwin (1969). Agarcontaining medium was incubated in an atmosphere of 5 % COZ in N2.Assessment of viable ureaplasmas. This was done by determining d o u r change units (c.c.u.) (Taylor-Robinson and Purcell, 1966;Gourlay et al., 1972), or by determining colonyforming units (c.f.u.).Mouse inocula. These were prepared either by centrifuging " overnight " broth cultures at lo00 g for 20 min. and resuspending the deposits in 0 . 1 5~ phosphate-buffered saline (PBS) pH 7.2, or by thawing from -70°C broth cultures containing a known number of c.f.u. and diluting them in PBS just before use to give the required number of organisms.Mice. BSVS mice bred at Compton were used, unless otherwise stated. Mice of C3H, CBA and TO strains were bred at the Clinical Research Centre.Infection and examination of mice. The technique used to inoculate mice by the intramammary route has been described previously (Chandler, 1970); the fourth gland from the front on each side (R4 and L4) was used and the volume of the inoculum was 0.0541 ml.

“…Other in vitro studies on normal bovine milk and whey demonstrated a dialysable factor capable of killing a variety of mycoplasma species, including ureaplasma and M. dispar (Brownlie, Howard & Gourlay, 1974;Howard et at. 1975).…”

Section: Discussionmentioning

confidence: 99%

“…The Ureaplasma sp., strain Vic 9, had previously been shown to induce mastitis in cows (Howard, Gourlay & Brownlie, 1973) and likewise the M. dispar strain Gri 226 (Brownlie et al 1976). The ureaplasma was grown in urea-broth (Howard et at. 1975) and M. dispar in glucose-serum broth (Gourlay & Leach, 1970) containing ampicillin (Andrews et at.…”

Section: Mycopla8masmentioning

confidence: 99%

The effect of an intramammary infusion of endotoxin on experimentally induced mycoplasmal mastitis

Brownlie

Howard

1979

J. Hyg.

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The infusion of 10 microgram of endotoxin lipopolysaccharide from Escherichia coli into the mammary gland of four cows 16 h before inoculation with ureaplasmas did not prevent, or even diminish, the subsequent ureaplasmal mastitis. There was no reduction in the severity or duration of the inflammatory cell response in milk or in the clinical appearance of the resulting mastitis. Also, the excretion of ureaplasmas was not reduced. A similar experiment with Mycoplasma dispar in two cows demonstrated that endotoxin was again ineffective in preventing the mastitis. Furthermore, there was some indication that the proliferation and excretion of this mycoplasma was enhanced in endotoxin-treated quarters.