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A F Angulo,
R Reijgers,
J Brugman,
I Kroesen,
F E Hekkens,
P Carle,
J M Bové,
J G Tully,
A C Hill,
L M Schouls,
C S Schot,
P J Roholl, A A Polak-Vogelzang
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ABSTRACT: Organisms isolated from commercial foetal bovine serum and from cell culture lines containing such serum supplements were found to consist of non-helical, non-motile, pleomorphic coccoid forms. One strain (FC 097-2T) cultivated directly from foetal bovine serum was selected for characterization. In ultrastructural examination, individual round cells lacked cell wall structures and cells varied in size, with a mean diameter of about 700 nm. However, variable numbers of cells were filterable through membranes of 300 nm. Optimum growth occurred between 30 and 37 degrees C. The organism fermented glucose, fructose and mannose, but did not hydrolyse arginine. The strain was insensitive to 500 U penicillin ml(-1) and was capable of growing in the absence of serum or cholesterol. The organism was serologically distinct from all 13 currently described species in the genus Acholeplasma and from other sterol-requiring species in the genus Mycoplasma, using growth inhibition, immunoperoxidase and immunofluorescence tests. Strain FC 097-2T was found to have a DNA G+C composition between 37.6 +/- 1 mol% and 38.3 +/- 1 mol%. The genome size was determined to be 2095 kbp. The 16S rDNA sequence of strain FC 097-2T was compared to 16S rDNA sequences of other mollicutes in nucleotide databases. No deposited sequence was found to be identical; the closest relatives were several members of the genus Acholeplasma. On the basis of these findings and other similarities to acholeplasmas in morphology and growth, the absence of a sterol requirement for growth, and similar genomic characteristics, the organism was assigned to the genus Acholeplasma. Strain FC 097-2T is designated the type strain (ATCC 700667T) of a new species, Acholeplasma vituli.
International journal of systematic and evolutionary microbiology 06/2000; 50 Pt 3:1125-31. · 2.27 Impact Factor
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ABSTRACT: Acholeplasma strains were isolated from the nasopharynx of a horse (strain PN525T [T = type strain]) and the feces of a rabbit (strain B1). One clone of strain PN525T and one clone of strain B1 were examined in detail. These clones were indistinguishable from each other and were serologically distinct from the previously described Acholeplasma and Mycoplasma spp. The strains had the following properties: guanine-plus-cytosine content of 31 mol%; sterol was not required for growth, which occurred under both aerobic and anaerobic conditions; glucose was metabolized; and arginine was hydrolyzed. Strain PN525 (= NCTC 11723) is the type strain of a new species, Acholeplasma multilocale.
International journal of systematic bacteriology 11/1992; 42(4):513-7.
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ABSTRACT: The survival of four strains of Mycoplasma hyorhinis in stock solutions of trypsin was tested at 22, 4 and -15 degrees C. Low (10(4)-10(5) cfu/ml) and high (10(6)-10(7) cfu/ml) initial concentrations of each strain were used, each was tested three times. A regular decrease of low and high concentrations (1 log in 10 and 20 min, respectively) was seen at 22 degrees C. At 4 degrees C the low concentrations showed a reduction of about 1 log/h, while apart from one strain high concentrations hardly decreased during the first 6 h and the survival time ranged from 24 to more than 30 h at the end of which there was a reduction of 4 logs. At -15 degrees C low concentrations survived up to 1 week in only one of the three tests, high concentrations survived for more than 12 weeks (reduction 3 logs). These latter results suggest that mycoplasmas may be present in trypsin as clumps, which deteriorate very slowly. A study was also performed to compare the sensitivity of different cultural procedures for detecting mycoplasmas.
Biologicals 05/1990; 18(2):97-101. · 1.70 Impact Factor
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ABSTRACT: Acholeplasma laidlawii was isolated from the faeces of 23.5% and 24% of groups of 51 conventional and 45 specified-pathogen-free (SPF) rabbits respectively. Isolation of the organism from individual animals could often be repeated, suggesting that infection was not merely transient. Two further acholeplasmas were isolated from two SPF rabbits. One was serologically related to Acholeplasma modicum. The other could not be identified and may be a new species.
Laboratory Animals 08/1987; 21(3):201-4. · 1.21 Impact Factor
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ABSTRACT: Heifers were immunized against Mycoplasma arginini, M. fermentans, M. hyorhinis and M. orale and the antisera were applied for elimination of these species from cell cultures. From fifteen out of nineteen contaminated human and animal cell cultures the mycoplasmas could be eliminated by treating the cells with medium with 10% or 20% antiserum (eight cases) or antiserum combined with one or two antibiotics (six cases). In ten cases two treatments were sufficient, in four cases respectively four, six or eight (2 X) treatments were necessary, in one case antiserum combined with a heat treatment (42 degrees C) was successful. The efficacy of the treatment depended on the antibody titer of the serum, the contaminating mycoplasm species (M. arginini being more difficult to eliminate than the other three species) and the cells involved. The bovine sera were not cytotoxic, except for a slight toxicity for a mouse lymphoma cell line. The application of specific bovine antiserum for elimination of mycoplasmas is an easy and often successful method.
Zentralblatt für Bakteriologie, Mikrobiologie, und Hygiene. Series A, Medical microbiology, infectious diseases, virology, parasitology 05/1987; 264(1-2):84-92.
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ABSTRACT: Two strains of Lewis rat were successfully freed from Mycoplasma pulmonis infection by using a combination of oral treatment with oxytetracycline hydrochloride and obtaining young by hysterectomy. Laminar flow cabinets were used to perform hysterectomies on donor animals and for rearing hysterectomy-derived animals. After thorough microbiological examination the rats were brought to the breeding colony of the Laboratory Animal Centre. Periodic laboratory tests using both cultural and enzyme-linked immunosorbent assay methods showed that the animals have remained free from M. pulmonis for the last 3 years.
Laboratory Animals 04/1987; 21(2):138-42. · 1.21 Impact Factor
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ABSTRACT: A total of 1949 cell cultures was tested for contamination with mollicutes by cultivation on and in mycoplasma media, 25.7% of the cell cultures was positive, 243 strains of Mycoplasma hyorhinis were isolated. Furthermore, mainly M. arginini and M. orale were detected, less often Acholeplasma laidlawii, M. fermentans and M. pneumoniae. Optimal conditions for isolation were discussed. About one third of 217 hybridoma cultures and two third of 57 myeloma cultures proved to be contaminated, all with M. hyorhinis. A DNA fluorochrome staining method (DAPI-test) was compared to cultivation for testing 1039 cell cultures. The efficiency of the DAPI-test could be estimated to be about 96% that of cultivation about 89%, but cultivation is more specific. The highest assurance is obtained when both methods are applied.
Antonie van Leeuwenhoek 02/1987; 53(2):107-18. · 2.09 Impact Factor
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Zeitschrift für Versuchstierkunde 02/1986; 28(3):123-8.
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ABSTRACT: The first isolations of Mycoplasma pulmonis were made from inflamed ovaries of 2 C3H/F1 mice. Investigation of cultures from a further 110 apparently healthy mice revealed 14 cases of M. pulmonis localized in the ovaries and associated with oophoritis.
Laboratory Animals 11/1985; 19(4):275-6. · 1.21 Impact Factor
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ABSTRACT: The efficiency of aerobic incubation was compared with incubation under various oxygen and carbon dioxide conditions for the isolation and subcultivation of three strains of Mycoplasma hyorhinis from VERO-cell cultures and subcultivation of three laboratory strains. Under anaerobic conditions with a low oxidation-reduction potential (at or below -115 mV) as obtained in jars, with catalysts, containing mixtures of 5%-10% CO2 in H2, very poor or no growth of any of the six M. hyorhinis strains was observed. When traces of oxygen were present (that is, under conditions with higher oxidation-reduction potentials, e.g. when omitting the catalyst in the above gas mixtures or in 5% CO2 + 95% N2) isolation from cell cultures was successful in most tests, but subcultivation of these primary isolates was seldom possible under these semi-anaerobic conditions. However, in most cases these primary isolates could be subcultivated aerobically, although aerobic conditions were unsatisfactory for isolation in about half of the experiments. Isolation of M. hyorhinis was optimal in 5% O2 + 95% N2, under which condition the isolates could also always be subcultivated. Isolation failed occasionally when 5% O2 + 5% CO2 + 90% N2 was used, thus indicating that 5% CO2 was slightly inhibitory. 5% CO2 in air and 10% CO2 either in air, H2 or N2 were also inadequate for isolation from cell cultures. In contrast to the findings with these cell culture-adapted M. hyorhinis strains, the laboratory strains could be subcultivated easily under all conditions tested except those with an oxidation-reduction potential at or below -115 mV; 100% CO2 was inhibitory for all 6 strains. Our findings may partly explain why M. hyorhinis is often considered "non-cultivable" on artificial media once adapted to cell cultures. The findings emphasize the need to employ also a micro-aerophilic condition (5% O2 in 95% N2) in the examination of cell cultures for mycoplasma.
Antonie van Leeuwenhoek 05/1983; 49(1):31-40. · 2.09 Impact Factor
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Journal of Biological Standardization 02/1981; 9(3):277-85.
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Journal of Biological Standardization 02/1980; 8(4):243-54.
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A A Polak-Vogelzang
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ABSTRACT: The survival of two strains of Mycoplasma gallispeticum in mains water at room temperature depended on the initial number of organisms and the incorporation of mycoplasma broth in the suspending medium. Seven to 8 log(10) colony forming units (cfu) ml were needed for survival for 1 day in sterile water. In water with 1% (vol/vol) broth at least 6 log(10 )cfu/ml were needed for survival, which was prolonged to 4 days. In water with 10% broth 1 to 4 log(10) cfu/ml could survive for 9 to 10 days, which is similar to the survival time of low concentrations of mycoplasma in pure broth.
Avian Pathology 02/1977; 6(1):93-5. · 1.71 Impact Factor
