The Isolation and Comparative Biological and Physical Characteristics of T-mycoplasmas of Cattle

1991

A mycoplasma isolated from human spermatozoa and a human cervix was shown to be serologically distinct from 98 previously recognized MycopEasma and AchoZepZasma spp. Six mycoplasma colonies were cloned and examined in detail for morphology, growth, and biochemical characteristics; five of these were from sperm samples and one was from a cervix. These strains were closely related and had the following properties: guanine-plus-cytosine content of 32 mol% , requirement for sterol, and anaerobic growth. Glucose was not metabolized, and arginine and urea were not hydrolyzed. Strain AH159 (= NCTC 11720) is the type strain of a new species, Mycoplasma spermatophilum.Twelve named Mycoplasma and Acholeplasma species have been isolated from the respiratory or genital tracts of humans (6). Mycoplasma buccale, Mycoplasma faucium, Mycoplasma lipophilum , Mycoplasma orale, Mycoplasma pneumoniae, and Mycoplasma salivarium are found almost exclusively in respiratory tracts. Mycoplasma fermentans has been found infrequently in urogenital tracts, while Mycoplasma primatum, a species commonly present in nonhuman South American primates (15), has on occasion been isolated from humans. Acholeplasma laidlawii, which is also uncommon in humans, has been isolated from many animal species and plants (26).The more common human genital mycoplasmas are Ureaplasma urealyticum and Mycoplasma hominis, although each of these species has also been found in human respiratory systems. Both of these species have been implicated in certain diseases (e.g., nongonococcal urethritis and pelvic inflammatory disease) (30). The association of ureaplasmas with some other diseases has not been proved as controls are often similarly infected. It is possible that serotyping of ureaplasmas may clarify this position.Recently, a fastidious mycoplasma was isolated from human genital tracts by using SP4 medium (37). This mycoplasma, Mycoplasma genitalium was recovered from patients with nongonococcal urethritis (36), but the respiratory tract is probably the primary host site of colonization (3).Strains 9,47,50,58, AH159T (T = type strain), and Monks were isolated from the urogenital tracts of patients with infertility problems and could not be identified as any of the previously described human species. In this paper I describe the characteristics of these organisms and propose that they are members of a new Mycoplasma species. MATERIALS AND METHODSMycoplasma strains. Mycoplasmas were isolated from the spermatozoa of five patients attending an in vitro fertilization clinic (16) and from the cervix of one woman attending a clinic for infertility (strain Monks). The cervix isolate was obtained from a cervical swab rubbed over the surface of an agar culture medium. Semen and washed sperm were inoculated onto the agar medium and SP4 agar and into glucose, arginine, and urea broth media and SP4 liquid medium (13,32,37). Most of the inoculated liquid media were diluted 1/10; the one exception was urea medium, which was diluted 10-fold to lo6. The amount of inoculum de...

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Section: Methodsmentioning

confidence: 99%

Mycoplasma spermatophilum, a New Species Isolated from Human Spermatozoa and Cervix

1991

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“…Mycoplasmas were isolated from the vagina of 10 apparently healthy guinea pigs in several different colonies during routine microbiological screening. Swabs taken from the vagina were rubbed over the surface of an agar culture medium and inoculated into glucose or arginine broth medium (11,29). The inoculated liquid medium was diluted 1/10.…”

Section: Methodsmentioning

confidence: 99%

Acholeplasma cavigenitalium sp. nov., Isolated from the Vagina of Guinea Pigs

1992

A mollicutes isolated from a guinea pig vagina was shown to be serologically distinct from previously recognized Mycoplasm and Acholeplasma species. Colonies isolated from 10 different guinea pigs were cloned and examined in detail. These strains were closely related and had the following properties: guanine-pluscytosine content of 36 mol%, no requirement for sterol, and aerobic growth. Glucose was not metabolized, and arginine and urea were not hydrolyzed. Strain GP3 (= NCTC 11727) is the type strain of a new species, Acholeplasma cavigenitalium.A mollicutes that could not be identified as one of those previously recovered from guinea pigs, i.e., Mycoplasma caviae, M. cavipharyngis, M. pulmonis, Acholeplasma granularurn, and A. laidlawii, was isolated from the vagina of guinea pigs (12,13,15). Strains were recovered from the vagina of 10 guinea pigs from several different colonies. In this paper, I describe the characteristics of these strains in comparison with those of strains of known genera and species and propose that they are members of a new Acholeplasma species. MATERIALS AND METHODSMycoplasma strains. Mycoplasmas were isolated from the vagina of 10 apparently healthy guinea pigs in several different colonies during routine microbiological screening. Swabs taken from the vagina were rubbed over the surface of an agar culture medium and inoculated into glucose or arginine broth medium (11,29). The inoculated liquid medium was diluted 1/10. One colony isolated from each guinea pig was cloned to produce a pure culture; this was done by filtering a broth culture through a 220-nm-pore-size membrane filter (Millipore Corp., Bedford, Mass.) by use of a syringe with slight pressure, culturing the filtrate on solid medium, transferring a single resulting colony to another plate, and inoculating the subsequent growth into broth. This entire procedure was repeated four times (27) Medium and growth conditions. The culture medium used was described previously (11, 29) and contained thallium acetate and penicillin. The purified agar used in the growth medium was either Ionagar no. 2 (Oxoid Ltd., Basingstoke, England) or Lab M agar (Lab M, London, England). Depending on their biochemical activities, the mycoplasmas were grown in liquid medium containing 1% (wtkol) glucose (pH 7.8) or in liquid medium containing 0.5 to 1% (wtkol) arginine (pH 7.3). The species grown in medium containing arginine were not inhibited by 1% arginine. Agar cultures were incubated at 35 to 37°C either in a humid chamber or under anaerobic conditions in a GasPak system. Liquid cultures were stored at -70°C.Growth requirements and characteristics. The clones were subcultured onto solid medium and incubated under aerobic conditions at 35 to 37°C. The levels of susceptibility to methylene blue were investigated by adding 0.002% (wtkol) methylene blue to the basal solid medium and comparing the growth of inoculated mycoplasmas with the growth of mycoplasmas cultivated on agar medium without methylene blue. Lipolytic activity was tested by inocula...

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“…Mycoplasmas were isolated from the throats of captive lions and a leopard; these animals were kept at the Zoological Society of London in Regents Park and Longleat Safari Park in the United Kingdom. Swabs taken from a throat were rubbed over the surface of an agar culture medium and were inoculated into broth media containing glucose and arginine (13,29). The inoculated liquid media were diluted 1/10.…”

Section: Methodsmentioning

confidence: 99%

Mycoplasma simbae sp. nov., Mycoplasma leopharyngis sp. nov., and Mycoplasma leocaptivus sp. nov., Isolated from Lions

1992