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PubMed1974

NYC medium for simultaneous isolation of Neisseria gonorrhoeae, large-colony mycoplasmas, and T-mycoplasmas.

NYC medium, primarily designed for isolation of pathogenic Neisseria, also readily supports the growth of large-colony mycoplasmas and T-mycoplasmas. When compared with A-3 agar medium and PPLO agar in clinical field trials, NYC medium performed equally as well as these mycoplasma-specific media in providing recovery of Mycoplasma species from female genital specimens. The transparent, highly-selective NYC medium permits direct, microscopic observation and presumptive identification of mycoplasmas, as well as Neisseria gonorrhoeae, without interference from contaminating saprophytes. As a single medium it can therefore be doubly useful in the diagnosis of gonorrhea and in the recognition of active or asymptomatic mycoplasma infections. Used in gonorrhea screening programs, the medium can be valuable in establishing the frequency of association of mycoplasmas with urogenital tract infection.

Bacteriological Techniques↗
PubMed1981

A preliminary study on isolation and enumeration of mycoplasmas in dental plaques: the effect of sonication on viability of oral mycoplasmas.

The effect of sonication on the viability of Mycoplasma salivarium and Mycoplasma orale was examined as a preliminary investigation on the quantitative studies on mycoplasmas in the dental plaques. Mycoplasma cell aggregations suspended in PBS or liquid medium were sonicated and the samples were removed at 5-second intervals for 20 seconds to check the viable counts. Inactivation of the mycoplasmas was almost directly proportional to the sonication time. M. orale was more sensitive to sonication than M. salivarium. Mycoplasmas were damaged more readily in PBS than in the liquid medium. Dental plaques suspended in PBS were also sonicated exactly as mentioned above. The number of mycoplasmas in the dental plaques (1 mg) ranged from 1.09 x 10(2) to 1.75 x 10(5) cfu. The desirable sonication time to prepare the homogeneous suspensions of dental plaques was concluded to be 5 seconds.

Dental Plaque↗
PubMed2005

Swine and poultry pathogens: the complete genome sequences of two strains of Mycoplasma hyopneumoniae and a strain of Mycoplasma synoviae.

This work reports the results of analyses of three complete mycoplasma genomes, a pathogenic (7448) and a nonpathogenic (J) strain of the swine pathogen Mycoplasma hyopneumoniae and a strain of the avian pathogen Mycoplasma synoviae; the genome sizes of the three strains were 920,079 bp, 897,405 bp, and 799,476 bp, respectively. These genomes were compared with other sequenced mycoplasma genomes reported in the literature to examine several aspects of mycoplasma evolution. Strain-specific regions, including integrative and conjugal elements, and genome rearrangements and alterations in adhesin sequences were observed in the M. hyopneumoniae strains, and all of these were potentially related to pathogenicity. Genomic comparisons revealed that reduction in genome size implied loss of redundant metabolic pathways, with maintenance of alternative routes in different species. Horizontal gene transfer was consistently observed between M. synoviae and Mycoplasma gallisepticum. Our analyses indicated a likely transfer event of hemagglutinin-coding DNA sequences from M. gallisepticum to M. synoviae.

Animals↗
PubMed1990

Comparative in vitro activity of azithromycin, clarithromycin, erythromycin and lomefloxacin against Mycoplasma pneumoniae, Mycoplasma hominis and Ureaplasma urealyticum.

The in vitro activity of three macrolides, azithromycin, clarithromycin and erythromycin and a new fluoroquinolone, lomefloxacin, against pathogenic mycoplasma (16 to 18 strains of Mycoplasma pneumoniae, 41 to 77 strains of Mycoplasma hominis, 65 to 104 strains of Ureaplasma urealyticum) was compared. The three macrolides were highly active against Mycoplasma pneumoniae. Clarithromycin was the most active macrolide against Ureaplasma urealyticum whereas azithromycin was somewhat more active than erythromycin against Mycoplasma hominis. Lomefloxacin was moderately active against all three mycoplasma species.

4-Quinolones↗
PubMed1989

Binding of thyrotropin to selected Mycoplasma species: detection of serum antibodies against a specific Mycoplasma membrane antigen in patients with autoimmune thyroid disease.

Radiolabeled human (hTSH) and bovine (bTSH) thyroid stimulating hormone was shown to bind to five species of Mycoplasma, the wall-less prokaryotes. The maximum binding capacity of 125I-bTSH to these five species was about 7.9 x 10(-13) moles-1.4 x 10(-12) moles for 50-100 micrograms protein with dissociation constants of approximately 1.7 to 2.2 x 10(-7)M. Approximately 50% of the 125I-bTSH binding was displaced by excess, unlabeled bTSH or hTSH, but labeled bTSH was not effectively displaced by growth hormone, LH, FSH, prolactin, or the beta subunit of hTSH, FSH and LH. Antisera prepared against Mycoplasma gallisepticum and Mycoplasma pneumoniae bound to human thyroid membranes and guinea pig fat cells, suggesting that receptors on human thyroid tissues and on Mycoplasma cells may have similarities in antigenicity. These findings were substantiated by the occurrence of TSH binding to Mycoplasma antisera. Further, sera from three of six patients with Graves' disease containing antibodies to thyroid tissues also reacted to a 108 Kd polypeptide of Mycoplasma gallisepticum.

Animals↗
PubMed1993

Models of infection due to mycoplasmas, including Mycoplasma fermentans, in the genital tract and other sites in mice.

Progesterone induces susceptibility in murine genital tracts to Mycoplasma pulmonis, Mycoplasma pneumoniae, and Mycoplasma genitalium when each is given intravaginally. In contrast, estradiol, but not progesterone, induced such susceptibility to Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma fermentans, the "incognitus" and PG18 strains of the latter behaving similarly. None of the mice that had been given M. fermentans intravaginally died, whereas others that had been given large numbers of M. fermentans organisms intraperitoneally died within a few days. M. pulmonis spreads from the respiratory tract to other sites more often in T cell-depleted mice than in immunocompetent mice. In addition, the "incognitus" strain of M. fermentans given intravenously killed BALB/c nude mice but did not kill immunocompetent control mice of the same strain or CBA nude mice. The effect is not unique to M. fermentans since M. hominis inoculated intravenously also killed CBA nude mice or made them ill. Such observations raise the possibility that immunosuppression caused by the human immunodeficiency virus could allow mycoplasmas to flourish and exacerbate disease.

Animals↗
PubMed1978

Canine mycoplasmas: pathogenicity of mycoplasmas associated with distemper pneumonia.

Mycoplasmas of the following species were isolated from the lungs of dog with distemper: Mycoplasma cynos, Mycoplasma bovigenitalium, Mycoplasma canis, Mycoplasma spumans, and Mycoplasma gateae. One-week-old dogs were inoculated endobronchially with cloned cultures of each species. M. cynos induced a focal pneumonia that were characterized histologically by severe inflammation of bronchi and adjacent respiratory tract tissue. The inflammatory response was initially neutrophilic and later was histiocytic with interstitial lymphoid hyperplasia. M. cynos was recovered from the lungs of inoculated dogs until three weeks after inoculation. A mild inflammatory response was observed in lung tissue of the dog inoculated with M. bovigenitalium, whereas M. canis, M. spumans, and, M. gateae did not induce any visible lesions.

Animals↗
PubMed2000

The association of Mycoplasma hominis, Ureaplasma urealyticum and Mycoplasma genitalium with bacterial vaginosis: observations on heterosexual women and their male partners.

The prevalence of 3 mycoplasmas (Mycoplasma hominis, Ureaplasma urealyticum and Mycoplasma genitalium) was determined in a cohort of women with or without bacterial vaginosis (BV) and in their respective male partners. Heterosexual women with or without BV and their male partners were recruited and genital sampling for these microorganisms was performed. Seventeen women with BV and 21 women with normal flora, and their respective male partners, were recruited. M. hominis was present in 9 (53%) of 17 women with BV compared with none of 21 women without BV (P=0.0001). Of the 17 male partners of women with BV, 8 (47%) had M. hominis compared to 5 (24%) of 21 male partners of women without BV (not significant [n/s]). U. urealyticum was detected in 11 (65%) of 17 women with BV in comparison with 10 (48%) of 21 women without BV (n/s). U. urealyticum was present in 4 (24%) of 17 male partners of women with BV compared to 6 (29%) of 21 male partners of women without BV (n/s). M. genitalium was not detected in any of 15 women with BV and in only 2 (12%) of 17 women without BV (n/s). M. genitalium was present in 4 (25%) male partners of 16 women with BV in comparison with 3 (16%) male partners of 19 women without BV (n/s). Thus, M. hominis was the only mycoplasma detected significantly more often in women with, rather than in those without, BV. None of the mycoplasmas was found significantly more often in male partners of women with, rather than those without, BV. Overall, M. genitalium behaved somewhat similar to Chlamydia trachomatis. It was the least commonly occurring mycoplasma, a reflection perhaps of the relatively low incidence of partner change in this study population.

Carrier State↗
PubMed1986

Experimental studies on the pathogenicity of Mycoplasma ovipneumoniae and Mycoplasma arginini for the respiratory tract of goats.

Mycoplasma ovipneumoniae and Mycoplasma arginini were the species of Mollicutes most commonly isolated from 175 goats with respiratory disease in Ontario. The pathogenicity of M. ovipneumoniae, strain B321B and M. arginini, strain D53e, was assessed in goats following endobronchial inoculation. One out of three two year old goats developed fever after inoculation with a pure culture of strain B321B, and it had extensive subacute fibrinous pleuritis when necropsied three weeks later. Neither of the remaining goats had lesions in the respiratory tract. Mycoplasma ovipneumoniae was recovered from one of the animals four days after inoculation, but not at necropsy from any of the goats, at which time a marked humoral immune response with growth inhibiting antibodies was detected. In a second experiment three four to five week old goats were inoculated with the same strain and three other goats were given placebo treatment. One experimental goat developed fever and coughing, and it had extensive subacute fibrinous pleuritis in the right side and pneumonia. Another goat had focal pneumonia in the left diaphragmatic lobe. Microscopically there was subacute hyperplastic suppurative bronchiolitis, atelectasis and nonsuppurative alveolitis. The infected animals did not clear the mycoplasma and not all of them produced antibodies. Mycoplasma arginini, strain D53e, did not induce lesions in any of four goat kids within 14 days after inoculation but did cause transient elevations in rectal temperature, circulating monocytes, circulating neutrophils and blood fibrinogen. Mycoplasma arginini was infective and immunogenic for all inoculated animals and showed a particular affinity for the tonsil. Thus, this study provides the first evidence that M. ovipneumoniae is pathogenic for goats causing pneumonia and pleuritis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Genetic and antigenic relatedness between Mycoplasma gallisepticum and Mycoplasma synoviae.

The two avian pathogens Mycoplasma gallisepticum and Mycoplasma synoviae were found, by Southern blot hybridization of their digested DNAs, to share genomic nucleotide sequences additional to those of the highly conserved ribosomal RNA genes. The assumption that some of the shared sequences encode for antigens or epitopes common to both mycoplasmas was supported by Western immunoblot analysis of cell proteins of one mycoplasma with specific antiserum to the other mycoplasma. Interestingly, the band patterns of reactive antigens were different for some of the M. gallisepticum strains, supporting the concept that the species is genotypically variable. The results of the present study may explain the cross-reactivity of the two mycoplasmas noted previously in a variety of routine serological tests.

Animals↗
PubMed1982

Polyamines in mycoplasmas and in mycoplasma-infected tumour cells.

Three out of four different mycoplasma strains analysed for the polyamine contents contained relatively high concentrations of putrescine, cadaverine, spermidine and spermine. In addition to ornithine decarboxylase (EC 4.1.1.17) activity, the mycoplasmas also exhibited comparable or higher lysine decarboxylase (EC 4.1.1.18) activity fully resistant to the action of 2-difluoromethylornithine, an irreversible inhibitor of eukaryotic ornithine decarboxylase. 2-Difluoromethylornithine did not modify the polyamine pattern of actively growing mycoplasmas. Ehrlich ascites carcinoma cells and L1210 mouse leukemia cells infected with any of the four mycoplasma strains contained, in addition to putrescine, spermidine and spermine, and also easily measurable concentrations of cadaverine; the latter diamine was absent in uninfected cultures. When the infected cells were exposed to difluoromethylornithine, the accumulation of cadaverine was markedly enhanced. The modification of cellular polyamine pattern by mycoplasmas, especially in the presence of inhibitors of eukaryotic ornithine decarboxylase, could conceivably be used as an indicator of mycoplasma infection in cultured animal cells.

Animals↗
PubMed1993

Mycoplasma imitans sp. nov. is related to Mycoplasma gallisepticum and found in birds.

A mycoplasma designated strain 4229T (T = type strain) was isolated in 1984 from the turbinate of a duck in France, and similar strains were isolated from geese in France and from a partridge in England. All of these strains were originally identified as Mycoplasma gallisepticum by immunofluorescence and growth inhibition tests, but subsequent serological and molecular studies indicated only a partial relationship to this species and DNA-DNA hybridization studies revealed only approximately 40 to 46% genetic homology with M. gallisepticum PG31T. In this study morphological, cultural, and physical investigations were carried out on strain 4229T and partridge strain B2/85, after we first demonstrated the similarity between these organisms by performing a restriction enzyme analysis of their DNAs. Both strains had phenotypic properties very similar to M. gallisepticum properties, including the presence of an attachment organelle. As a result of these investigations, the organisms were assigned to the class Mollicutes, the order Mycoplasmatales, and the genus Mycoplasma. They fermented glucose, reduced triphenyl tetrazolium chloride aerobically and anaerobically, and exhibited hemadsorption and hemagglutination, but other biochemical tests were negative. Apart from a serological cross-reaction with M. gallisepticum, these organisms exhibited no significant relationship with any previously described Mycoplasma species as determined by growth inhibition or immunofluorescence tests or with a number of additional serovars and unclassified avian strains. This Mycoplasma taxon therefore appears to be a new species, for which we propose the name Mycoplasma imitans. The type strain is strain 4229 (= NCTC 11733 = ATCC 51306).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2004

Spreading factors of Mycoplasma alligatoris, a flesh-eating mycoplasma.

Mycoplasma alligatoris causes lethal invasive disease of alligators and caimans. A homolog of the nagH gene, encoding a hyaluronidase secreted by Clostridium perfringens, and a C. perfringens hyaluronidase nagI or nagK pseudogene were discovered in the M. alligatoris genome. The nagH gene was detected by PCR in the closest relative of M. alligatoris, Mycoplasma crocodyli, but not in 40 other species representing the Mycoplasma hominis, Mycoplasma pneumoniae, and Spiroplasma phylogenetic clusters. The hyaluronidase activity in the cellular fraction of M. alligatoris and M. crocodyli SP4 broth cultures was equivalent to 10(-16) U of Streptomyces hyalurolyticus hyaluronidase CFU(-1). Negligible activity was present in the cell-free supernatant fraction. No chondroitinase activity was detected. There is also a novel homolog of the nanI gene, which encodes a sialidase secreted by C. perfringens, in the M. alligatoris genome. The signature YRIP and SXDXGXTW motifs and catalytic residues of the clostridial sialidase are conserved in the mycoplasmal gene, but the leader sequence necessary for its secretion by C. perfringens is absent. The gene was not detected by PCR in any other mycoplasma. Potent cell-associated sialidase activity was present in M. alligatoris colonies on agar but not in the cell-free supernatants of broth cultures or in M. crocodyli. The presence of hyaluronidase and sialidase in M. alligatoris is consistent with the rapid invasiveness and necrotizing effects of this organism, and the lack of sialidase in M. crocodyli is consistent with its comparatively attenuated virulence. This genetic and biochemical evidence suggests that the spreading factors hyaluronidase and sialidase, a combination unprecedented in mycoplasmas, are the basis of the virulence of M. alligatoris.

Alligators and Crocodiles↗
PubMed1966

Pneumonia due to mycoplasma in gnotobiotic mice. II. Localization of Mycoplasma pulmonis in the lungs of infected gnotobiotic mice by electron microscopy.

Organick, Avrum B. (Marquette University School of Medicine, Milwaukee, Wis.), Kenneth A. Siegesmund, and Irving I. Lutsky. Pneumonia due to mycoplasma in gnotobiotic mice. II. Localization of Mycoplasma pulmonis in the lungs of infected gnotobiotic mice by electron microscopy. J. Bacteriol. 92:1164-1176. 1966.-Lesions in lungs of gnotobiotic mice inoculated intranasally with Mycoplasma pulmonis were examined by electron microscopy after osmic acid fixation. At 1 week after infection, mycoplasma cells were found in large numbers in the bronchi at the surface of bronchial epithelial cells and, in smaller numbers, in the alveoli where active phagocytosis by polymorphonuclear leukocytes (PMN) occurred. Cytopathic changes in underlying bronchial epithelial cells, not apparent by light microscopy, were observed. At 3 weeks after infection, mycoplasma cells were rarely seen in the bronchi, and were no longer seen free in the alveolar spaces or within PMN. Lungs examined after glutaraldehyde fixation 1 week after infection confirmed the presence of mycoplasma cells in the alveolar spaces and within phagocytic vacuoles of PMN, but also revealed numerous ring forms within granular pneumocytes. These forms seemed to represent intracytoplasmic developmental stages of M. pulmonis, in which elementary bodies appeared in large numbers.

Animals↗
PubMed1981

Attachment of mycoplasmas to erythrocytes: a model to study mycoplasma attachment to the epithelium of the host respiratory tract.

Mycoplasma pneumoniae, M. gallisepticum and M. pulmonis are pathogens of the respiratory tract. Their adherence to the host tissue is a prerequisite for manifestation of the disease. The attachment occurs between membrane components of the mycoplasmas and the host cells. In the attachment of M. pneumoniae and M. gallisepticum, binding sites of a protein nature of the mycoplasma membranes interact primarily with sialic acid residues of sialoglycoproteins of the host cell membranes. The latter was clearly indicated in studies on the pathogenic strain of M. pneumoniae and its nonpathogenic nonhemadsorbing mutants, and by the assessment of the direct binding of glycophorin [the major sialoglycoprotein of human red blood cells (RBC)] and other sialoglycoproteins to M. pneumoniae and M. gallisepticum. The components involved in the interaction of M. pulmonis with the host cells are not as well characterized, since the attachment is not affected by proteolytic treatment of the mycoplasmas, nor by removal of sialic acid residues from the host cell membranes. The attachment of the mycoplasmas to the host cells also occurs with nonviable organisms and can, in addition, by reproduced by isolated membranes. The binding sites of M. pneumoniae and M. gallisepticum were partially purified by affinity chromatography, using the high affinity of the binding sites for glycophorin. By this procedure the membranes were solubilized with detergents and chromatographed through glycophorin attached to Sepharose 4B as an affinity matrix. The binding sites retained the high affinity for glycophorin and bound to RBC membranes. Are the binding sites distributed throughout the membrane? Since the binding of the mycoplasma is via a polar structure (the tip or bleb), one may suggest that these structures harbor them. Experimental evidence for their distribution is being sought.

Adhesiveness↗
PubMed1993

Adhesion onto and invasion into mammalian cells by mycoplasma penetrans: a newly isolated mycoplasma from patients with AIDS.

The newly identified mycoplasma, Mycoplasma pentrans shows remarkable pathobiologic properties: it adheres to cell surfaces, deeply penetrates into the cell, strongly hemadsorbs human red blood cells, and cytadsorbs human CD4+ lymphocytes and monocytes. These in vitro biologic activities of mycoplasmas have been previously shown to be associated with pathogenic virulence in vivo. Both adhesion and invasion clearly involve the organism's unique tip-like structure. Invading mycoplasmas often have their tip-like structure deeply buried in the cytoplasm of infected mammalian cells. Extensive invasion of the mycoplasma into the cytoplasm may kill the cells. The same pathobiologic processes of adhesion and invasion using the specialized tip-like structure are found on the epithelium in the patient's urogenital tract infected by M. penetrans. Both in vitro and in vivo findings suggest a possible pathogenic role of this newly discovered human mycoplasma and call for careful evaluation of its role in human diseases.

Acquired Immunodeficiency Syndrome↗
PubMed1993

Use of a triplex polymerase chain reaction for the detection and differentiation of Mycoplasma pneumoniae and Mycoplasma genitalium in the presence of human DNA.

PCR primers corresponding to the adhesin genes of Mycoplasma pneumoniae and Mycoplasma genitalium were shown to detect the corresponding organisms specifically. Absence of cross-reaction with seven other mollicute species and six unrelated bacterial species commonly found in humans was demonstrated. Positive control primers directed against human mitochondrial DNA could be mixed with the Mycoplasma primers without loss of specificity or sensitivity. A detection level of 10 c.f.u. of either Mycoplasma species could be readily obtained, even in the presence of 10(4) human cells. The triplex PCR method developed is very simple and does not require hybridization or the use of radioisotopes and allows detection and differentiation of these mycoplasmas against the background of human DNA found in clinical specimens.

Base Sequence↗
PubMed1989

Interaction of Mycoplasma bovis and Mycoplasma bovigenitalium with preimplantation bovine embryos.

In vitro exposures of bovine embryos to Mycoplasma bovis and Mycoplasma bovigenitalium were conducted to determine if these organisms adhered to the zona pellucida-intact (ZP-I) bovine embryo, and standard procedures for washing and treating embryos were evaluated to determine their effectiveness for removing or killing mycoplasmas. Mycoplasma bovis and M. bovigenitalium were isolated from 19 of 19 and 24 of 24 ZP-I embryos, respectively, after in vitro exposure and subsequent washing, thus demonstrating adherence of the two species of Mycoplasma to the ZP. Additionally, M. bovis was isolated from 20 of 20 and 23 of 23 embryos, while M. bovigenitalium was isolated from 25 of 25 and 22 of 22 embryos after antibiotic and trypsin treatment, respectively. It was concluded that neither of the standard procedures currently used for cleansing embryos should be relied upon for insuring freedom from mycoplasmas.

Journal Article↗