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PubMed1989

Incidence of Mycoplasma hominis and Ureaplasma urealyticum infection in pregnant women and gynaecological patients; the effectivity of doxycycline therapy.

The authors report on the results of Mycoplasma hominis and Ureaplasma urealyticum cultivations performed within two and a half years in 1,421 pregnant women and gynaecological patients. Considering the obstetrical-gynaecological patients Mycoplasma hominis was identified in 7.25% and Ureaplasma urealyticum in 38.11% of cases. No significant difference was found in the incidence of Mycoplasma infection when comparing the results of cultivations of samples obtained from a clinically non-inflamed vagina or cervix with the results of cultivations of samples obtained from the female lower genital tract in the course of an inflammatory disease. In cases of endometritis, salpingitis or cophoritis Mycoplasma hominis could be isolated from the cervix significantly more frequently. The incidence of Mycoplasma hominis infection was also significantly higher in the examined 315 sterile women. The incidence of Mycoplasma hominis and Ureaplasma urealyticum infection did not differ when comparing the 267 symptom-free pregnant women to the other patients. The presence of the pathogenic organisms did not aggravate the unfavourable course of pregnancy. When examining the effectivity of Doxycycline capsule the drug proved to be effective in 78 patients for the treatment of genital Mycoplasma infections.

Doxycycline↗

PubMed1988

Isolation and identification of mycoplasmas from the nasal cavity of sheep.

Mycoplasmas isolated from the nasal cavity of sheep in a ram test station were examined to determine their identity and prevalence. Specimens were obtained for mycoplasmal culture in 1980, 1982, and 1983 from 558 sheep, and mycoplasmas were isolated from 630 specimens from 320 sheep (57.3%). The isolates were characterized and differentiated into groups on the basis of sensitivity to digitonin, fermentation of glucose, and hydrolysis of arginine. Isolates in some groups were further characterized by use of additional diagnostic media, and their identity was confirmed by agglutination or growth inhibition with antiserum prepared from reference mycoplasmas. Of the 320 sheep with mycoplasmas, 293 had Mycoplasma ovipneumoniae, 12 had M arginini, and 1 had M capricolum. Two sheep had Acholeplasma spp, and 3 sheep had unidentified Mycoplasma spp. The remaining 9 sheep had M ovipneumoniae in combination with Acholeplasma spp (n = 3), M arginini (n = 3), M capricolum (n = 2), and an unidentified Mycoplasma spp (n = 1). The biochemical reactions of the M ovipneumoniae from the 293 sheep were similar, but varied in the degree of growth and fermentation in the basal medium containing glucose. The high prevalence of M ovipneumoniae indicated that it may be commensal in the upper respiratory tract of healthy sheep.

Animals↗

PubMed1984

Effect of the caprine variant of Mycoplasma mycoides subsp mycoides on endothelium, monocytes, and complement of guinea pig, calf, sheep, and goat serum.

The caprine variant of Mycoplasma mycoides subsp mycoides causes septicemia with coagulopathy in goats. Pathogenetic mechanisms that might explain the coagulopathy, the ability of the Mycoplasma to persist in the blood, and its specificity for goats were studied. Severe endothelial damage was seen by electron microscopy of goat aorta tissue exposed in vitro to 10(7) colony-forming units of mycoplasmas. The Mycoplasma did not damage 51Cr-labeled adherent cells from peripheral blood of goats. The hemolytic complement titer was reduced by 94%, 50%, 50%, and 25% in guinea pig, calf, sheep, and goat serum, respectively, 30 minutes after treatment with 8 X 10(9) colony-forming units of the Mycoplasma. Freshly prepared serum from these animal species killed the Mycoplasma. Heat-inactivated serum was not mycoplasmacidal. Complement from these 4 animal species was activated by the Mycoplasma through the classical pathway, because ethyleneglycoltetraacetic acid precipitation of serum Ca2+ inhibited activation. Proof that the classical pathway was functional in goats was not conclusive because Ca2+ supplementation of ethyleneglycoltetraacetic acid-treated serum did not restore complement activity. Endothelial damage and complement activation may explain the coagulopathy. The function that complement activation may have in the inflammatory response of this disease is not known. Difference in susceptibility of calves, sheep, and goats to M mycoides septicemia cannot be explained by species variation in complement mycoplasmacidal activity.

Animals↗

PubMed1984

Antibiotic sensitivities and elimination of mycoplasmas from infected cell cultures.

Invaluable or irreplaceable cell lines infected with mycoplasmas can be cured. We have had success with a protocol that utilizes a combination of anti-mycoplasma immune serum and pretested antibiotics. A panel of antibiotics was evaluated by examining inhibition of agar growth of a variety of cell culture mycoplasmas. Resistant strains were found with certain drugs and intraspecies heterogeneity was seen in drug response. For these reasons drug regimens must be tailored to individual strains. When possible, two effective antibiotics were chosen to minimize chances of outgrowth of drug-resistant subpopulations. Cell lines were diluted and cultured in treatment cocktail consisting of growth medium plus antibiotics and antiserum. After a short exposure, cells were subcultured into fresh growth medium. Routine culturing practices were resumed and mycoplasma monitoring was begun. This procedure has been effective in curing eight cell lines, of which five were monolayers and three were suspension cultures. Mycoplasmas from four species comprised the contaminants. Two cures were of mixed infection caused by strains of two mycoplasma species. No failures have been encountered with this procedure thus far. Cultures have remained free of mycoplasmas and have retained properties of interest.

Animals↗

PubMedDate not supplied

Species identification of avian mycoplasmas by polymerase chain reaction and restriction fragment length polymorphism analysis.

Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis were used to detect and differentiate four pathogenic species (Mycoplasma gallisepticum, M. iowae, M. meleagridis, and M. synoviae) and ten nonpathogenic species of avian mycoplasma. A sequence of 1026 base pairs within the gene for 16S ribosomal RNA (16S rRNA) from avian mycoplasmas was successfully amplified by PCR with oligonucleotide primers (M16SPCR5' and M16SPCR3') common to all avian mycoplasmas tested. Restriction endonucleases (REs) with unique restriction sites, selected by computer-assisted analysis of known sequences of the amplified segment of avian mycoplasma, were then used to digest the PCR products. After electrophoresis of the resulting RE fragments, the RFLP patterns were compared. Combinations of up to six REs (HpaI, HhaI, HaeIII, HphI, FokI, and NlaIV) produced unique RFLP patterns by which the 14 species of avian mycoplasmas could be differentiated. The newly classified avian species M. imitans was also investigated by this method; M. imitans and M. gallisepticum gave identical RFLP patterns with the REs used in this study. The results obtained by the PCR and RFLP analysis were in agreement with current methods for species identification of avian mycoplasmas.

Animals↗

PubMed1993

Factors influencing the ability of different mycoplasmas to colonize the genital tract of hormone-treated female mice.

Colonization of the genital tract of female mice, mainly BALB/c, by Mycoplasma genitalium, M. pneumoniae and M. pulmonis was enhanced by pretreatment of the mice with progesterone. M. fermentans, M. hominis and M. salivarium, and three serotypes and two untyped strains of Ureaplasma urealyticum colonized under the influence of oestradiol but not progesterone. Mycoplasmas dependent on progesterone were glucose-metabolizing, with strong haemadsorptive and other attachment properties, and possessed a terminal structure. Mycoplasmas dependent on oestradiol were arginine or arginine/glucose metabolizing. Ureaplasmas also required oestradiol. The oestradiol-requiring group appeared to be less cytadsorptive and devoid of a morphological terminal structure. Mycoplasmas that had had multiple passes in media were less able to colonize. This may be one, but not the only, reason for the failure of seven mycoplasmas to colonize under the influence of either hormone. The observations suggest the existence of a receptor mechanism for colonization, progesterone-requiring mycoplasmas being exposed to, and needing, genital tract cells different from those exposed to oestradiol-requiring mycoplasmas.

Animals↗

PubMedDate not supplied

[Presence of Mycoplasma in laboratory cell cultures from Cordoba, Argentina].

In this paper we determined the prevalence of mycoplasma contamination in 17 cell lines. Eighty per cent of the laboratories that currently use cell culture techniques participated in this study. Hoechst 33258 dye was used to detect mycoplasma contamination. The relationship between culture maintenance conditions and the presence of mycoplasma were analyzed, considering the use of antibiotics in the culture media, fetal calf serum (FCS) quality, culture media processing, use of disponsable labware, type of laminar flow cabinet, quantity of operators, and cell culture system. Thirty-five per cent of the analyzed cell lines showed mycoplasma contamination. Those lines belonged to 2 of the 8 surveyed laboratories. When confronting the working conditions versus mycoplasma contamination, 66% of the laboratories that employ non-certified FCS or reuse their labware, show mycoplasma contamination. Mycoplasma presence was found in 50% of the laboratories that use closed culture system, or more than one operator. Laboratories that process their culture media or that include antibiotic in the growing media, show a 40% contamination. The results obtained help to establish working conditions necessary to avoid introducing or spreading the microorganism.

Argentina↗

PubMed1998

Comparison of polymerase chain reaction assay with culture for detection of genital mycoplasmas in perinatal infections.

The polymerase chain reaction (PCR) technique was compared with culture for the detection of Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium in clinical samples (vaginal secretions, throat and endotracheal secretions, and skin swabs) obtained from 47 high-risk pregnant women peripartum and eight newborn infants. Detection using PCR with homologous primers was highly specific, as a product with the expected length was consistently amplified with homologous but not with heterologous species. The limit of detection of the PCR assay was 10 color-changing units (CCU) of Mycoplasma strains. The PCR technique facilitated detection of Ureaplasma urealyticum DNA in 31 of 55 patients studied, of Mycoplasma hominis in seven samples, and of Mycoplasma genitalium in two samples. Four PCR-positive patients yielded culture-negative results. In one case a culture-positive sample was negative by PCR. The results show that PCR is a valuable tool for rapid detection of genital mycoplasmas in clinical samples. It is fast, sensitive, specific, and easy to perform, requiring minimal preparation of the clinical sample.

Base Sequence↗

PubMed2006

Quantitative detection and phylogeny-based identification of mycoplasmas and ureaplasmas from human immunodeficiency virus type 1-positive patients.

More than 20 years ago, Mycoplasma genitalium was first isolated from men with nongonococcal urethritis (NGU). Some strains of Mycoplasma and Ureaplasma are associated with NGU or other genitourinary infections. It is reasonable to suggest that mycoplasmas and ureaplasmas may increase the development of acquired immunodeficiency syndrome in human immunodeficiency virus type 1 (HIV-1)-infected patients. This study investigated a rapid and reliable diagnostic method, based on a LightCycler SYBER green I real-time PCR assay and phylogenetic analysis, for detecting mycoplasmas and ureaplasmas in urine from HIV-1-infected patients. Primers were designed to amplify a 711-bp fragment of the 16S rRNA gene. Positive PCR-amplified products were further sequenced, and a phylogenetic tree was constructed by neighbor joining of the V4 and V5 sequences from 15 prototypes strains. PCR-positive samples were detected in 17 (9.1%) urine samples, their prototypes being as follows: 2/17 (11.2%) M. genitalium, 1/17 (5.9%) M. hominis, 12/17 (70.6%) Ureaplasma urealyticum, 1/17 (5.9%) mixed M. genitalium and M. hominis, and 1/17 (5.9%) M. hominis and U. urealyticum, respectively. Mycoplasma and Ureaplasma load for each specimen was 3.8 x 10 copies/ml to 7.2 x 10(3) copies/ml. Overall, this study demonstrates that real-time PCR and phylogenetic analysis are the fastest available methods for the detection of mycoplasmas in the urogenital tract.

Adult↗

PubMed2005

Prevalence survey on condom use and infection of urogenital mycoplasmas in female sex workers in China.

GOAL: We evaluated the prevalence of condom use and the effects of condom use on urogenital mycoplasma infection in female sex workers in Jinjiang, China. METHODS: Two-hundred ninety-nine female sex workers from Jinjiang city, Jiangsu Province, were interviewed, and three mycoplasmas of Ureaplasma urealyticum (Uu), Mycoplasma hominis (Mh) and Mycoplasma genitalium (Mg) were detected by nested polymerase chain reaction in genital secretions of 72 female sex workers and 42 female patients seen with symptoms of genital infection (control group). RESULTS: The results showed that 87.29% of the sex workers used condom in their last sexual intercourse, and that 2.68% did not use, 38.80% sometimes used and 58.52% used condoms every time in the last month. Those who were older, married and have a stable partnership used condoms less frequently. The infection rates of Uu, Mh and Mg were 77.78%, 34.72% and 16.67%, respectively, in sex worker subjects compared to those of controls at 59.52%, 9.62% and 21.43%, respectively. The infection rates of Uu and Mg were lower among all subjects (NS) who used condoms every time. In this observational study, genital mycoplasma infection was common and occurred more frequently among sex workers, and infection occurred less often when condoms were used. These results tend to support the efficacy of condom use in reducing urogenital mycoplasma infection among Chinese women.

Adolescent↗

PubMed1965

CYTOPATHOGENIC MYCOPLASMAS ASSOCIATED WITH TWO HUMAN TUMORS. II. MORPHOLOGICAL ASPECTS.

Hummeler, K. (The Children's Hospital of Philadelphia, Philadelphia, Pa.), D. Armstrong, and N. Tomassini. Cytopathogenic mycoplasmas associated with two human tumors. II. Morphological aspects. J. Bacteriol. 90:511-516. 1965.-Cytopathic effects (CPE) produced in HeLa cell cultures by two strains of mycoplasmas (F-11 and F-12) were studied by light and electron microscopy. CPE, which was marked by cytoplasmic vacuolization, did not appear to depend on intracellular mycoplasma infection. The cytopathogenic mycoplasmas appeared to be similar, both morphologically and in their intra- and extracellular distribution, to noncytopathogenic mycoplasmas previously studied by others. The probability that the CPE is related to depletion of essential nutrients is discussed, and the fine structure of the mycoplasmas is described.

Cytoplasm↗

PubMed1996

Search for the presence of six Mycoplasma species in peripheral blood mononuclear cells of subjects seropositive and seronegative for human immunodeficiency virus.

The prevalence of Mycoplasma fermentans, Mycoplasma pirum, Mycoplasma genitalium, Mycoplasma pneumoniae, Mycoplasma hominis, and Mycoplasma penetrans was investigated by using specific PCR assays with peripheral blood mononuclear cells from subjects infected or not infected with the human immunodeficiency virus (HIV). Only M. fermentans was detected in 5.8% of 154 HIV-seropositive and 11.1% of 90 HIV-seronegative subjects.

AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

Detection of mycoplasmas in urethral swabs from HIV-1 infected patients and control individuals using culture techniques and polymerase chain reaction.

The objective of the present study was to determine the prevalence of certain mycoplasma species, i.e., Mycoplasma hominis, Ureaplasma urealyticum and Mycoplasma penetrans, in urethral swabs from HIV-1 infected patients compared to swabs from a control group. Mycoplasmas were detected by routine culture techniques and by the Polymerase Chain Reaction (PCR) technique, using 16SrRNA generic primers of conserved region and Mycoplasma penetrans specific primers. The positivity rates obtained with the two methods were comparable. Nevertheless, PCR was more sensitive, while the culture techniques allowed the quantification of the isolates. The results showed no significant difference (p < 0.05) in positivity rates between the methods used for mycoplasma detection.

Acquired Immunodeficiency Syndrome↗

PubMed1975

The importance of mycoplasmas in respiratory infections.

In contrast to the uncertain role of mycoplasmas in genital disease, there is no doubt that they produce respiratory disease. The first mycoplasma isolated, M. mycoidesis is the aetiological agent of bovine pleuropneumonia, a disease now confined to parts of Africa and Australia. M. capri causes a similar disease in goats which is geographically more widespread. Enzootic pneumonia of pigs, seen in various parts of the world, is due primarily to M. suipneumoniae. Although some strains of M. hyorhinis also seem capable of causing disease. Not all strains of a mycoplasma species are equally pathogenic; only some strains of M. pulmonis produce severe respiratory disease in mice and rats. Further, concomitant infection with other agents may result in more severe disease; this is particularly seen in M. gallisepticum infections of poultry. Of those mycoplasmas which infect man, only M. pneumoniae is known to cause respiratory disease, mainly involving the younger age groups. Mycoplasmas spread by close contact so that disease is seen in family groups or where there is crowding or herding. Factors in resistance to respiratory disease caused by mycoplasmas have been poorly understood but the importance of local immunity is becoming more clear.

Adolescent↗

PubMed2001

[New type pathogenic mycoplasmas isolated from female patients with genital infection in Nanjing, China].

OBJECTIVE: To study if infection of new type pathogenic mycoplasmas, Mycoplasma penetrans (Mpe), Mycoplasma fermentans (Mf), and Mycoplasma pirum (Mpi), exists in the genital tract of gynecologic patients with genital infection in China so as to provide information for clinical diagnosis and therapy. METHODS: Cervical secretions from 172 patients with infectious diaseses of gentital tract and 172 healthy controls were collected and inoculated in improved SP-4 culture medium. Biochemical reaction, coagglutination test, metabolism-inhibition test, polymerase chain reaction (PCR) assay, and DNA sequencing were employed to identify the isolated microorganisms. RESULTS: One strain of Mycoplasma penetrans (0.58%) and two strains of Mycoplasma fermentans (1.16%) were isolated from the cervical secretions of patients, but none was found in healthy controls. CONCLUSION: Mpe infection and Mf infection exist in female patients with genital infection in Nanjing.

Adult↗

PubMed1990

Spontaneous bronchopneumonia in laboratory dogs infected with untyped Mycoplasma spp.

Three Mycoplasma spp. were isolated from five colony bred laboratory dogs (Canis familiaris) obtained from a single vendor. Four of these animals were Beagles and one was a mongrel. Three displayed clinical signs of respiratory disease including dyspnea, chronic coughing and moist rales, while the other two dogs were observed during thoracic surgery to have macroscopic lesions suggestive of pneumonia. All five dogs were submitted for diagnostic necropsy during which they were cultured for bacteria and mycoplasma. Mycoplasma spp. having three distinct colonial forms were isolated from the lungs of each of the animals. These three isolates were sent to the National Cancer Institute Diagnostic Microbiology Laboratory and to the National Institutes of Health, NIAID, Mycoplasmology Laboratory. Neither laboratory could serotype these isolates against antisera to 73 Mycoplasma spp., including the common canine mycoplasmas, and nine Acholeplasma spp. Histologically, the bronchopneumonia was characterized by bronchiectasis, purulent bronchiolitis, bronchial and bronchiolar epithelial hyperplasia, chronic non-suppurative peribronchiolitis and perivasculitis, bronchiolitis obliterans, and acute to subacute purulent pneumonia. The similarity between the pathologic findings in these animals and those observed in respiratory mycoplasmosis of other species, e.g. the rat, suggests a causal relationship between the isolated mycoplasmas and the pulmonary disease observed in these dogs.

Animals↗

PubMed1997

Molecular biology of Mycoplasma.

Mycoplasmas are the smallest free living microorganisms with the smallest genome. The G+C content is in general low (25-33%) and the coding capacity is about 600 proteins. Mycoplasma species are phylogenetically related, they use the genetic codon UGA for tryptophan, and show rapid evolution, with a high rate of divergence. The genomes of Mycoplasma genitalium and Mycoplasma pneumoniae have been fully sequenced. Striking features of the M. genitalium sequencing project are the presence of a high number of membrane proteins with no resemblance to previously sequenced genes and the presence of repeated fragments of the gene encoding the tip-localized 140 kDa adhesin (MgPa). Many Mycoplasma species display a high frequency of antigenic variation, both as phase and size variation of individual antigens. Mycoplasma hominis isolates are known to be antigenic heterogeneous, as reflected in the reactivity with monoclonal antibodies (MAbs). The genetics of the antigenic variation has been studied for three different surface exposed antigens: P120, Lmp, and P50/Vaa. The gene encoding P120 had a hyper-variable region in the N-terminal region. In addition, a second gene with homology to p120 was identified. The gene encoding Lmp, a 135 kDa protein is repeated and both genes are translated and both contain internal repeated sequences. Deletion mutants in the lmp gene were obtained by cultivation of M. hominis PG21 with MAb 552 specific for the repeated part of Lmp. One of the lmp genes had deletions of from four to eight repeats. The other gene was left unaltered. The genes encoding P50/Vaa show a different form of variability where domains of the genes seem to be exchangeable. The genomic maps of five M. hominis strains showed that even though the size of the genomes varied the position of the different genes were in general conserved.

Animals↗

PubMed1966

Genetic differentiation by nucleic Acid homology I. Relationships among Mycoplasma species of man.

Reich, Paul R. (National Institutes of Health, Bethesda, Md.), Norman L. Somerson, Carol J. Hybner, Robert M. Chanock, and Sherman M. Weissman. Genetic differentiation by nucleic acid homology. I. Relationships among Mycoplasma species of man. J. Bacteriol. 92:302-310. 1966.-Genetic relatedness among human mycoplasmas was evaluated by measuring the amount of nucleic acid hybrid retained on a membrane filter. Hybrids were formed from deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) derived from representative strains of seven serologically distinct human Mycoplasma species. The results indicate that serologically distinct human Mycoplasma species can also be distinguished by the homology techniques. Low-level cross-reactivity was observed among nucleic acids derived from the seven species. Genetic heterogeneity was demonstrated among three strains of M. salivarium and between two strains of M. orale type 2. In contrast, comparison of three strains and three passage levels of M. pneumoniae revealed them to be indistinguishable. M. pneumoniae appears to be the most distinct of all human mycoplasmas, as shown by both homology and the high buoyant density value of its DNA. Nucleic acids from mycoplasmas which had identical buoyant densities were in some cases differentiable. Mycoplasmas with different DNA buoyant densities were invariably distinguishable by the homology technique.

Journal Article↗