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1.  Isolation of Ureaplasma diversum and mycoplasmas from genital tracts of beef and dairy cattle in Saskatchewan 
We report herein a survey in which cultures of bovine reproductive tracts for Ureaplasma diversum and mycoplasmas were carried out in order to better understand the role of these organisms in granular vulvitis (GV). Samples cultured were vulvar swabs from clinically normal cows or ones with GV, preputial swabs or raw semen from bulls, and abomasal contents of aborted fetuses.
Ureaplasma diversum was isolated from 104 (43.3%) of 240 dairy cows, 32 (27.1%) of 118 beef cows, 43 (47.2%) of 91 beef heifers, 23 (67.6%) of 34 beef bulls, and three (60%) of five dairy bulls. Mycoplasmas were isolated from 18 (7.5%) dairy cows, two (1.6%) beef cows, three (8.8%) beef bulls, and one dairy bull. No isolation was made from 97 aborted fetuses. For 65 dairy cows and 30 beef heifers with vulvar lesions, the isolation rates for ureaplasmas of 62.5% and 69.7%, respectively, were significantly higher (X2) than those for normal animals (37.5% and 30.3%). On immunofluorescent serotyping of 137 of the 205 isolates, there were 66 in serogroup C (strain T44), 18 in serogroup B (strain D48), eight in serogroup A (strain A417 or strain 2312), 14 cross-reacting, and 31 that were not identified. It was concluded that U. diversum is commonly present in the lower reproductive tract of beef/dairy cattle in Saskatchewan and is associated with granular vulvitis.
PMCID: PMC1481172  PMID: 17423929
2.  Ureaplasma urealyticum, Ureaplasma parvum, Mycoplasma hominis and Mycoplasma genitalium infections and semen quality of infertile men 
Genital ureaplasmas (Ureaplasma urealyticum and Ureaplasma parvum) and mycoplasmas (Mycoplasma genitalium and Mycoplasma hominis) are potentially pathogenic species playing an etiologic role in both genital infections and male infertility. Reports are, however, controversial regarding the effects of these microorganisms infections in the sperm seminological variables. This study aimed at determining the frequency of genital ureplasmas and mycoplasmas in semen specimens collected from infertile men, and at comparing the seminological variables of semen from infected and non-infected men with these microorganisms.
A total of 120 semen samples collected from infertile men were investigated. Semen specimens were examined by in-house PCR-microtiter plate hybridization assay for the presence of genital ureaplasmas and mycoplasmas DNA. Semen analysis was assessed according to the guidelines of the World Health Organization. Standard parametric techniques (t-tests) and nonparametric techniques (Wilcoxon tests) were used for statistical analysis.
The frequency of genital ureaplasmas and mycoplasmas detected in semen samples of infertile men was respectively 19.2% (23/120) and 15.8% (19/120). The frequency of Ureaplasma urealyticum (15%) was higher than that of Mycoplasma hominis (10.8%), Ureaplasma parvum (4.2%) and Mycoplasma genitalium (5%). Mixed species of mycoplasmas and ureaplasmas were detected in 6.7% of semen samples.
Comparison of the parameters of the standard semen analysis between the male partners of the infertile couples with and without genital ureaplasmas and mycoplasmas infection showed that the presence of Mycoplasma hominis DNA in semen samples is associated with low sperm concentration (p = 0.007) and abnormal sperm morphology (p = 0.03) and a negative correlation between sperm concentration and the detection of Mycoplasma genitalium in semen samples of infertile men (p = 0.05). The mean values of seminal volume, pH, vitality, motility and leukocyte count were not significantly related either to the detection of genital mycoplasmas DNA or to the detection of ureaplasmas DNA in semen specimens.
Our results demonstrate that genital mycoplasmas and ureaplasmas seem to be widespread among the male partners of infertile couples in Tunisia. Genital mycoplasmas infections of the male genital tract could negatively influence semen quality. Our results also indicate that PCR-microtiter plate hybridization assay method provides a rapid and effective technique to detect human genital mycoplasmas and ureaplasmas which is useful for etiological and epidemiological studies of these pathogens.
PMCID: PMC2194714  PMID: 17988404
3.  The Distribution of Mycoplasmas and Ureaplasmas in the Genital Tract of Normal Artificial Insemination Bulls 
Bull semen is commonly contaminated with mycoplasmas. To determine the source of contamination, semen and the genital tracts of 45 artificial insemination bulls were cultured for these organisms. The results indicate that mycoplasmas colonize the prepuce and the distal part of the urethra. Only rarely were they found in the ampullae or seminal vesicles. In 92% of the bulls with contaminated semen the same Mycoplasma species or Ureaplasma diversum was isolated from the prepuce and urethral orifice as was found in the semen. This suggests that the prepuce and distal urethra is the source of contamination. Colonization of the genital tracts with Mycoplasmas or U. diversum was not associated with histological changes.
PMCID: PMC1680053  PMID: 17422489
Mycoplasma; ureaplasma; genital tract; semen; artificial insemination; microbial colonization
4.  Campylobacter fetus in artificial insemination unit and slaughterhouse bulls in Ontario. 
Preputial fluid samples were collected from 90 bulls in two Ontario artificial insemination units using a penial glove swab technique previously developed by one of us for use in donor bulls. No Campylobacter fetus organisms were identified from the prepuce or from samples of semen collected at the same time from these bulls. The distal genitalia of 200 bulls were collected at a slaughter house. One isolation of a Campylobacter fetus subspecies venerealis was obtained on a culture from the fornix area of the prepuce of one of these bulls.
PMCID: PMC1236155  PMID: 3893662
5.  The genital Mycoplasma and Ureaplasma flora of healthy and diseased dogs. 
The genital mycoplasma and ureaplasma flora was compared in 136 dogs with varied reproductive histories. Mycoplasmas were recovered from 88% of vulvovaginal swabs, 85% preputial swabs and 72% semen samples. Isolation rates were slightly higher from dogs that were infertile or had evidence of genital disease but the differences from those that were fertile or clinically normal were statistically significant only in the male. Ureaplasmas were recovered from half the females sampled. Higher, but not statistically significant isolation rates (75%) were made from infertile females with purulent vulvar discharge versus those that were clinically normal and fertile (40%). In the male dog there was a significantly higher incidence of ureaplasmas in the prepuce of infertile animals (69%) than those that were fertile (0%) (p less than or equal to 0.05). Semen isolations although not significantly higher in infertile males, were all made from ejaculates, with subnormal motility, low sperm counts and/or a high percentage of midpiece and tail abnormalities (bent or tightly coiled).
PMCID: PMC1320213  PMID: 7340908
6.  Genital Mycoplasmas in Placental Infections 
Objective: The involvement of the genital mycoplasmas Ureaplasma urealyticum and Mycoplasma hominis in complications of pregnancy has remained controversial especially because these microorganisms are frequent colonizers of the lower genital tract. Recovery of bacteria from the placenta appears to be the sole technique to represent a true infection and not vaginal contamination. Therefore, we investigated the presence of genital mycoplasmas, aerobic and anaerobic bacteria, and fungi in human placentas and evaluated their association with morbidity and mortality of pregnancy.
Methods: We cultured placentas from 82 women with complicated pregnancies. One hundred placentas from women with uncomplicated pregnancies were evaluated as controls. When possible, placentas were examined histologically for presence of chorioamnionitis.
Results: Microorganisms were recovered from 52% of the placentas of complicated pregnancies and U. urealyticum was the microorganism isolated most frequently from the placenta. A significant association between positive mycoplasma culture of the placenta and complication of pregnancy was found, and chorioamnionitis was positively related to isolation of mycoplasmas.
Conclusions: These data suggest that genital mycoplasmas are able to infect the human placenta where they can cause chorioamnionitis. This infection of the placenta by genital mycoplasmas is related to preterm birth and fatal outcome of pregnancy.
PMCID: PMC2364351  PMID: 18475351
7.  Comparison of Multiplex PCR Assay with Culture for Detection of Genital Mycoplasmas 
Journal of Clinical Microbiology  2004;42(4):1528-1533.
Ureaplasma, spp. Mycoplasma genitalium, and Mycoplasma hominis are associated with infection of the genitourinary tract, reproductive failure, and neonatal morbidity and mortality. We have developed a multiplex PCR for the detection of these Mycoplasmatales in a single amplification reaction. The analytical sensitivities of this assay were 10.8, 10.8, and 8.8 CFU for each organism, respectively. This multiplex PCR was compared to culture on 26 cervical swabs, 2 vaginal swabs, 4 female urine specimens, 49 semen samples, 2 male urine specimens, and 1 nonspecified sample. A total of 21 specimens were culture positive (25%); 17 of these were PCR positive. An additional 11 specimens were PCR positive but culture negative. Of the 21 culture-positive specimens, 17 (81%) grew Ureaplasma spp. and 4 (19%) grew Mycoplasma spp. Of the 28 PCR-positive specimens, Ureaplasma spp. was detected in 23 (82%), M. hominis was detected in 3 (11%), and both were detected in 2 (7%). In a confirmatory analysis, all samples were tested by amplification of a second target of the ureaplasma genome. True-positive cases were defined as a positive result by culture or by both amplification assays. The multiplex PCR detected organisms in 26 of the 30 true-positive specimens, as well as in 2 other specimens. Based on a 36% prevalence of infection, the sensitivity, specificity, and positive and negative predictive values of multiplex PCR analyses were 87, 96, 94, and 93%, respectively. Multiplex PCR offers a rapid, sensitive, and easy method to detect genital mycoplasmas.
PMCID: PMC387538  PMID: 15070999
8.  Ureaplasma serovars & their antimicrobial susceptibility in patients of infertility & genital tract infections 
Background & objectives:
Ureaplasmas have been implicated in a variety of clinical conditions. However, only certain serovars of ureaplasmas are disease associated. Only a few classes of antimicrobial agents are available for the treatment of mycoplasmal infections in humans. Increase of resistance of genital mycoplasmas to antimicrobials has been reported worldwide. The aim of the present study was to determine the occurrence of Ureaplasma serovars in patients with infertility and genital tract infections with polymerase chain reaction (PCR)–based serotyping. The antimicrobial susceptibilities of Ureaplasma spp. and Mycoplasma hominis were also assessed to determine the most suitable treatment strategy.
Sexually active adults (n=147) with symptoms of genital tract infections and 115 infertile women were enrolled. Endocervical swabs from women and urethral swabs from men were subjected to culture and multiplex PCR for detection of genital mycoplasmas. Serotyping of Ureaplasma was done by PCR and antimicrobial susceptibility to doxycycline, azithromycin, josamycin and ofloxacin was done by microbroth dilution method.
Ureaplasma was detected in 25.8 per cent patients with genital tract infections and 20.8 per cent in infertile women. Serovar 3/14 was the most frequent isolate followed by serovar 1 and serovar 6. The majority of Ureaplasma isolates were susceptible to doxycycline (91%) and josamycin (86%) followed by ofloxacin (77%) and azithromycin (71%). All the isolates of M. hominis were uniformly susceptible to doxycycline, josamycin and ofloxacin.
Interpretation & conclusions:
The predominance of Ureaplasma serovar 3/14 suggests their possible pathogenic role in genital tract infections and infertility. For empirical treatment, doxycycline could be the drug of choice for genital mycoplasmas.
PMCID: PMC3612329  PMID: 23391795
Antimicrobial susceptibility; PCR; ureaplasma serovars
9.  Bovine Genital Mycoplasmosis 
The Canadian Veterinary Journal  1981;22(11):339-343.
Infection, lesions and clinical significance of Acheloplasmas, Mycoplasma bovis and Mycoplasma bovigenitalium in genital disease of cattle are described. A more detailed account is given of ureaplasma infections. Acute and chronic forms of granular vulvitis in both field and experimental disease are described as well as the role of the organism in abortion.
Recovery rates of ureaplasma and mycoplasma from semen and preputial washings in bulls are outlined and their significance in disease is discussed. There are problems in differentiating pathogenic from nonpathogenic isolates. Methods are being developed to treat semen for these organisms.
This paper provides a concise summary of clinical and microbiological aspects of bovine genital mycoplasmosis.
PMCID: PMC1789999  PMID: 7337908
10.  Antimicrobial susceptibility patterns of Ureaplasma species and Mycoplasma hominis in pregnant women 
BMC Infectious Diseases  2014;14:171.
Genital mycoplasmas colonise up to 80% of sexually mature women and may invade the amniotic cavity during pregnancy and cause complications. Tetracyclines and fluoroquinolones are contraindicated in pregnancy and erythromycin is often used to treat patients. However, increasing resistance to common antimicrobial agents is widely reported. The purpose of this study was to investigate antimicrobial susceptibility patterns of genital mycoplasmas in pregnant women.
Self-collected vaginal swabs were obtained from 96 pregnant women attending an antenatal clinic in Gauteng, South Africa. Specimens were screened with the Mycofast Revolution assay for the presence of Ureaplasma species and Mycoplasma hominis. The antimicrobial susceptibility to levofloxacin, moxifloxacin, erythromycin, clindamycin and tetracycline were determined at various breakpoints. A multiplex polymerase chain reaction assay was used to speciate Ureaplasma positive specimens as either U. parvum or U. urealyticum.
Seventy-six percent (73/96) of specimens contained Ureaplasma spp., while 39.7% (29/73) of Ureaplasma positive specimens were also positive for M. hominis. Susceptibilities of Ureaplasma spp. to levofloxacin and moxifloxacin were 59% (26/44) and 98% (43/44) respectively. Mixed isolates (Ureaplasma species and M. hominis) were highly resistant to erythromycin and tetracycline (both 97% resistance). Resistance of Ureaplasma spp. to erythromycin was 80% (35/44) and tetracycline resistance was detected in 73% (32/44) of Ureaplasma spp. Speciation indicated that U. parvum was the predominant Ureaplasma spp. conferring antimicrobial resistance.
Treatment options for genital mycoplasma infections are becoming limited. More elaborative studies are needed to elucidate the diverse antimicrobial susceptibility patterns found in this study when compared to similar studies. To prevent complications in pregnant women, the foetus and the neonate, routine screening for the presence of genital mycoplasmas is recommended. In addition, it is recommended that antimicrobial susceptibility patterns are determined.
PMCID: PMC3976045  PMID: 24679107
Pregnant women; Ureaplasma spp.; Mycoplasma hominis; Antimicrobial susceptibilities
11.  Animal model of Mycoplasma fermentans respiratory infection 
BMC Research Notes  2013;6:9.
Mycoplasma fermentans has been associated with respiratory, genitourinary tract infections and rheumatoid diseases but its role as pathogen is controversial. The purpose of this study was to probe that Mycoplasma fermentans is able to produce respiratory tract infection and migrate to several organs on an experimental infection model in hamsters. One hundred and twenty six hamsters were divided in six groups (A-F) of 21 hamsters each. Animals of groups A, B, C were intratracheally injected with one of the mycoplasma strains: Mycoplasma fermentans P 140 (wild strain), Mycoplasma fermentans PG 18 (type strain) or Mycoplasma pneumoniae Eaton strain. Groups D, E, F were the negative, media, and sham controls. Fragments of trachea, lungs, kidney, heart, brain and spleen were cultured and used for the histopathological study. U frequency test was used to compare recovery of mycoplasmas from organs.
Mycoplasmas were detected by culture and PCR. The three mycoplasma strains induced an interstitial pneumonia; they also migrated to several organs and persisted there for at least 50 days. Mycoplasma fermentans P 140 induced a more severe damage in lungs than Mycoplasma fermentans PG 18. Mycoplasma pneumoniae produced severe damage in lungs and renal damage.
Mycoplasma fermentans induced a respiratory tract infection and persisted in different organs for several weeks in hamsters. This finding may help to explain the ability of Mycoplasma fermentans to induce pneumonia and chronic infectious diseases in humans.
PMCID: PMC3544566  PMID: 23298636
Animal model; Mycoplasma; Mycoplasma fermentans; Respiratory infection
12.  Comparison of the new Mycofast Revolution assay with a molecular assay for the detection of genital mycoplasmas from clinical specimens 
BMC Infectious Diseases  2013;13:453.
Genital mycoplasmas are opportunistic bacteria that are associated with undesirable gynaecologic and reproductive events. Mycoplasmas are fastidious bacteria with increasing resistance to routine antimicrobials and often fail to grow on conventional culture methods. The commercial Mycofast Revolution assay permits the phenotypic detection and identification of genital mycoplasmas. Antimicrobial susceptibility testing against five antimicrobial agents with MICs corresponding to the CLSI guidelines can also be performed. This study aimed to compare the new commercially available Mycofast Revolution assay with a multiplex PCR assay.
Self-collected swabs were obtained from pregnant women attending the antenatal clinic of a tertiary academic hospital in Pretoria, South Africa from October 2012 to November 2012. These swabs were used to seed UMMt and modified Amies transport media. The seeded UMMt transported medium was used to inoculate the Mycofast Revolution assay for the identification, enumeration and antimicrobial susceptibility testing of genital mycoplasmas. Following DNA extraction from the modified Amies transport medium, specimens were subjected to a multiplex PCR assay for the detection of genital mycoplasmas.
The Mycofast Revolution kit had a sensitivity and specificity of 77.3% (95% CI: 62.15% to 88.51%) and 80% (95% CI: 28.81% to 96.70%), respectively, against the PCR assay. The positive and negative predictive values were 97.1% (95% CI: 85.03% to 99.52%) and 28.6% (95% CI: 8.57% to 58.08%). Genital mycoplasmas were detected in 71.4% (35/49) of samples with the Mycofast Revolution assay with 49% (24/49) being Ureaplasma spp. and 22.4% (11/49) mixed strains. The multiplex PCR assay had a positivity rate of 89.8% (44/49) for genital mycoplasmas; mixed strains were present in 51% (25/49) of samples, Ureaplasma spp. in 16.3% (8/49) and M. hominis in 22.4% (11/49) of samples.
There was a fair agreement (κ = 0.319) between the Mycofast Revolution assay and the mPCR assay. With the high prevalence rates of genital mycoplasmas, fast and efficient diagnostic methods are imperative to treat infections and minimise complications. The Mycofast Revolution assay is simple to use, has a short turn-around time and interpretation of results are straightforward. This assay circumvents common problems experienced with conventional culture and molecular methods in diagnostic laboratories where skilled personnel are limited and can be used as an alternative diagnostic assay.
PMCID: PMC3849776  PMID: 24079603
Mycoplasma hominis; Ureaplasma spp; Mycofast; Antimicrobial susceptibilities; Multiplex PCR assay
13.  Evaluation of PPLO, A7B, E, and NYC agar media for the isolation of Ureaplasma urealyticum and Mycoplasma species from the genital tract. 
Four agar media (PPLO, NYC, A7B, and E) which are commonly used for the isolation of urogenital tract Mycoplasma species and Ureaplasma urealyticum were compared by culturing swabs of the endocervix of 334 pregnant women on all four media. To permit growth of both Mycoplasma and U. urealyticum, selective ingredients were omitted from the media tested. A7B and E agar were both satisfactory for the isolation of Mycoplasma species, recovering 92 and 82%, respectively, of all Mycoplasma species isolated. Only A7B agar was satisfactory for U. urealyticum isolation, recovering 96% of all isolates. Several modifications of PPLO, NYC, and E agar failed to significantly improve recovery of U. urealyticum on these media. A7B agar was clearly superior to all other media tested in terms of recovery rate, typical appearance of colonies, and ease of reading. A7B can be used for the isolation of both U. urealyticum and Mycoplasma species from urogenital sites.
PMCID: PMC270982  PMID: 6690468
14.  Presence of Chlamydia, Mycoplasma, Ureaplasma, and Other Bacteria in the Upper and Lower Genital Tracts of Fertile and Infertile Populations 
Objective: The genital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) and Chlamydia trachomatis have been implicated as possible etiologic factors in infertility. Their role in patients with infertility needs to be further defined.
Methods: Seventy-nine infertile patients underwent laparoscopy with cultures obtained for aerobic and anaerobic bacteria, Chlamydia, Mycoplasma, and Ureaplasma from the peritoneal fluid, fallopian tube, endometrium, and endocervix. Cultures for similar organisms were taken from the endocervix of 80 fertile women in their first trimester. Culture results were also compared according to ovulatory status and laparoscopic findings in the infertile group.
Results: There were no differences in the recovery of Ureaplasma (29% vs. 28%) or Chlamydia (4% vs. 0%) positive cervical cultures in the fertile and infertile groups, respectively. However, a significantly higher number of Mycoplasma positive cervical cultures (14% vs. 5%, P = 0.05) were found in the fertile group. Only two upper genital tract cultures were found to be positive (Ureaplasma).
Conclusions: Therefore, if these organisms play a role in infertility, they are present and eradicated prior to infertility work-up and thus do not supports the use of a routine trial of antibiotics prior to laparoscopy.
PMCID: PMC2364301  PMID: 18475323
15.  Ureaplasma serotypes associated with the bovine urogenital tract. 
One hundred and thirty-one ureaplasma isolates were tested using the immunoperoxidase system. Thirty-four were from semen, 34 from preputial washes of normal bulls and 63 were from vaginal swabs from herds experiencing infertility problems and/or vulvovaginitis. The serotypes from semen were T44 (12.1%), Bu2 (11.2%), D48 (2.8%), T315 (0.9%) and T288 (0.9%). Those from preputial washes were T44 (9.3%), Bu2 (8.4%), T288 (7.5%), D48 (0.9%) and T95 (0.9%). From vaginal swabs the serotypes were D48 (22.4%), Bu2 (10.3%), T45 (4.7%), T288 (3.8%) and T315 (1.9%).
PMCID: PMC1235978  PMID: 6365295
16.  Recovery of Mycoplasma spp. from the Reproductive Tract of the Mare during the Estrous Cycle 
The Canadian Veterinary Journal  1987;28(8):519-522.
The sites in the genital tract from which mycoplasmas could be recovered at various stages of the estrous cycle were studied in five Standardbred mares naturally infected with Mycoplasma. Mycoplasma equigenitalium and Mycoplasma subdolum were most frequently isolated from the clitoral fossa as compared to the vagina, cervix, and uterus. The lowest isolation prevalence was observed in the uterus. The recovery of Mycoplasma spp. from the clitoral fossa did not differ at any stage of the estrous cycle; however, recovery from the vagina, cervix, and uterus was variable during the cycle and more organisms were recovered on the day of ovulation than at any other time. From these results it was concluded that the clitoral fossa is the most likely “ecological niche” for Mycoplasma spp. in the mare. Ureaplasmas were not isolated.
PMCID: PMC1680594  PMID: 17422844
Equine Mycoplasmas; mare; estrous cycle; reproductive tract
17.  Characterization and Comparative Genital Tract Pathogenicity of Bovine Mycoplasmas 1 
Infection and Immunity  1970;2(1):101-104.
Sixteen bovine genital mycoplasmal isolates obtained from semen and prepuce of bulls and from aborted fetuses were compared physiologically and serologically with the Donetta strain (tentatively Mycoplasma agalactiae var. bovis), a known pathogen. All isolates were distinct from the Donetta organism. Four appeared to be saprophytes, and the remainder were placed in one group which could not be further separated by the biochemical or serological methods used. Two of the organisms in the latter group have been subsequently identified as M. bovigenitalium. Uterine infusion of broth cultures of four isolates into virgin heifers failed to produce clinical evidence of disease, and significant lesions were not present at necropsy. The mycoplasmas were recovered from cervicovaginal mucus of only three heifers, and never for more than 3 days postinfusion. Since the organisms were not recovered from any organs at necropsy, it appears that the mycoplasmas were incapable of surviving in the clinically normal virgin female reproductive tract.
PMCID: PMC415971  PMID: 16557786
18.  Mycoplasmas and ureaplasmas in patients with hypogammaglobulinaemia and their role in arthritis: microbiological observations over twenty years. 
Annals of the Rheumatic Diseases  1994;53(3):183-187.
OBJECTIVES--To study the occurrence of mycoplasmas and ureaplasmas in patients with hypogammaglobulinaemia and the relationship of these micro-organisms to septic arthritis. METHODS--Over a period of about 20 years, 53 men and 38 women with hypogammaglobulinaemia, most of whom were less than 50 years old, were examined clinically and microbiologically. Mycoplasmas and ureaplasmas were sought in the throat, urogenital tract and joints by standard cultural methods, although not consistently in the three sites of all patients. RESULTS--Arginine-hydrolysing mycoplasmas and ureaplasmas occurred with similar frequency in the sputum/throat of the hypogammaglobulinaemic patients, but no more often than might be expected in immunocompetent patients. Ureaplasmas, however, dominated in the urogenital tracts of both men and women, being found in 75% of vaginal specimens. Arginine-hydrolysing mycoplasmas occurred two to six times more frequently and ureaplasmas two to three times more frequently in urine specimens from hypogammaglobulinaemic patients than they did in such specimens from sex- and age-matched non-venereal disease, hospital patients or healthy subjects; these differences were statistically significant (p < 0.05). Enhanced mucosal colonisation probably increases the chance of spread to distant sites, such as the joints. Of the 91 patients, 21 (23%) had septic arthritis involving one or more joints. Mycoplasmas and/or ureaplasmas, but not bacteria, were isolated from the joints of eight (38%) of these patients. However, dissemination to joints apparently had not occurred in some despite the opportunity by virtue of mycoplasmal or ureaplasmal colonisation at a mucosal site. Sometimes antibiotic therapy failed clinically, and microbiologically and recommendations for management are outlined. CONCLUSIONS--Hypogammaglobulinaemic patients appear to be more susceptible to colonisation of mucous membranes, especially of the urogenital tract, with mycoplasmas and ureaplasmas than are immunocompetent individuals. These micro-organisms are responsible for about two fifths of the septic arthritides occurring in these patients.
PMCID: PMC1005283  PMID: 8154936
19.  A high Nugent score but not a positive culture for genital mycoplasmas is a risk factor for spontaneous preterm birth 
This study was performed to evaluate the relationship among the Nugent score for the diagnosis of bacterial vaginosis (BV), the results of vaginal fluid culture for genital mycoplasma, and the subsequent occurrence of preterm birth.
The Nugent score and culture for genital mycoplasmas were performed in vaginal fluid obtained from 977 pregnant women (gestational age 13–30 weeks). Vaginal samples were obtained with sterile cotton swabs. The relationship among the Nugent score, vaginal fluid culture results and the occurrence of spontaneous preterm birth was examined.
(1) Of the 977 women, 14% (137) had a Nugent score of ≥ 8; (2) The prevalence of a positive vaginal culture for genital mycoplasmas was 30% (288); Ureaplasma urealyticum was isolated in 252 (88%), Mycoplasma hominis in 9 (3%), and both in 27 (9%) women; (3) Cases with a Nugent score of ≥ 8 had a higher rate of a positive vaginal culture for genital mycoplasmas than those with the lower Nugent score (55% vs. 25%; p<0.001); (4) Women with a Nugent score of ≥ 8 had a significantly higher rate of spontaneous preterm birth <37 (10% vs. 4%), <34 (5% vs. 2%), and <32 (4% vs. 1%) weeks of gestation than those with the lower Nugent score (At each gestational age, p<0.05); (5) In contrast, a positive vaginal culture for genital mycoplasmas was not associated with an increased risk for spontaneous preterm birth; (6) Among patients with a positive culture and a Nugent score of ≥ 8, the frequency of spontaneous preterm delivery (<37 weeks) was 10% (7/72); (7) There was no difference in the incidence of spontaneous preterm delivery according to the results of vaginal culture in patients with a Nugent score of ≥ 8, as well as in those with a lower Nugent score.
A high Nugent score (≥ 8) for the detection of BV but not a positive vaginal culture for genital mycoplasmas is a risk factor for spontaneous preterm birth.
PMCID: PMC3540826  PMID: 19330704
Nugent score; bacterial vaginosis; genital mycoplasmas; Ureaplasma urealyticum; Mycoplasma hominis; preterm birth; prematurity
20.  Prolonged eradication of urogenital mycoplasmas after administration of tetracycline to men in the Antarctic. 
Meatal swabs were obtained at intervals over 1 year from 23 men in the Antarctic. A 5-day course of tetracycline was given to twelve of them. In retrospect it was found that the antibiotic had been received by two men who were harbouring ureaplasmas, one of whom also had M. hominis. After treatment, these organisms were not found in any of the swabs taken over the next year, except in a swab from one of the men following sexual contact after this time. One of the twelve men developed N.S.U. just before arriving in the Antarctic. He responded clinically to a shorter course of tetracycline and ureplasmas were not recovered from a meatal swab immediately thereafter. However, without further sexual contact, ureaplasmas and disease recurred about a month later. This time, after a 5-day course of tetracycline, disease was not seen, and ureaplasmas were not isolated, over the next year. In contrast, ureaplasmas were isolated consistently over a year from two men who were not given the antibiotic. The evidence strongly suggests that, under natural conditions, the most likely cause of mycoplasmas, particularly ureaplasmas, recurring in the genital tract after apparently adequate tetracycline therapy, is re-infection as a result of sexual re-exposure.
PMCID: PMC1045296  PMID: 1036463
21.  The Semen Microbiome and Its Relationship with Local Immunology and Viral Load in HIV Infection 
PLoS Pathogens  2014;10(7):e1004262.
Semen is a major vector for HIV transmission, but the semen HIV RNA viral load (VL) only correlates moderately with the blood VL. Viral shedding can be enhanced by genital infections and associated inflammation, but it can also occur in the absence of classical pathogens. Thus, we hypothesized that a dysregulated semen microbiome correlates with local HIV shedding. We analyzed semen samples from 49 men who have sex with men (MSM), including 22 HIV-uninfected and 27 HIV-infected men, at baseline and after starting antiretroviral therapy (ART) using 16S rRNA gene-based pyrosequencing and quantitative PCR. We studied the relationship of semen bacteria with HIV infection, semen cytokine levels, and semen VL by linear regression, non-metric multidimensional scaling, and goodness-of-fit test. Streptococcus, Corynebacterium, and Staphylococcus were common semen bacteria, irrespective of HIV status. While Ureaplasma was the more abundant Mollicutes in HIV-uninfected men, Mycoplasma dominated after HIV infection. HIV infection was associated with decreased semen microbiome diversity and richness, which were restored after six months of ART. In HIV-infected men, semen bacterial load correlated with seven pro-inflammatory semen cytokines, including IL-6 (p = 0.024), TNF-α (p = 0.009), and IL-1b (p = 0.002). IL-1b in particular was associated with semen VL (r2 = 0.18, p = 0.02). Semen bacterial load was also directly linked to the semen HIV VL (r2 = 0.15, p = 0.02). HIV infection reshapes the relationship between semen bacteria and pro-inflammatory cytokines, and both are linked to semen VL, which supports a role of the semen microbiome in HIV sexual transmission.
Author Summary
The classical paradigm of HIV infectivity centers on the blood HIV RNA viral load. However, while other fluid compartments such as semen and cerebrospinal fluid can have distinct viral loads from blood, the causes of localized HIV shedding are not fully understood. Since the semen viral load is an independent predictor of HIV transmission risk, it is critical to understand the local factors that trigger increased semen viral shedding in order to develop novel preventative strategies. Here, we evaluated the semen microbiome, bacterial load, and cytokine levels in 22 HIV-uninfected men who have sex with men (MSM) and in 27 HIV-infected MSM before and after initiation of antiretroviral therapy (ART). We found that HIV infection reduces semen microbiome biodiversity, which is restored with ART and immune reconstitution. We also found that semen bacterial load in untreated, HIV-infected men is associated with the levels of seven semen cytokines, relationships not seen in the uninfected controls. In particular, the cytokine IL-1b was uniquely correlated with both semen bacterial and viral load. Our findings support the interaction between semen microbiome and local immunology, and suggest that IL-1b could be a mechanism for semen microbiome to trigger semen viral shedding.
PMCID: PMC4110035  PMID: 25058515
22.  Mycoplasmosis: Serology of Infections in the Genital Tract of Bulls 
Infection and Immunity  1972;5(1):20-23.
An investigation was carried out to determine the extent to which serology may contribute a means of elucidating the possible etiological significance of the presence of Mycoplasma bovigenitalium in bulls with genital tract disease. Experimentally infected bulls showed a significant serological response with maximum titers of antibody as early as 12 days after inoculation as measured by the indirect hemagglutination test. The tetrazolium reduction inhibition test, even as modified, was less suitable because this method did not reveal antibodies in all inoculated animals. The indirect hemagglutination test revealed high titers of antibody in serum of most bulls from bull stations in Denmark and Luxembourg although young bulls were often serologically negative. It is concluded that indirect hemagglutination is useful in experimental work and also in estimating the incidence of infection with mycoplasmas in bulls from artificial breeding stations. For diagnostic purposes, use of the indirect hemagglutination test is largely restricted to young bulls and on condition that the first blood sample is drawn very early in the course of the disease.
PMCID: PMC422313  PMID: 4656354
23.  Mycoplasmas isolated from the respiratory tract of horses. 
The Journal of Hygiene  1975;74(3):385-407.
Ten mycoplasmas were isolated from 130 nasopharyngeal swabs from thoroughbred horses with acute respiratory disease and three from 198 apparently normal horses. Two mycoplasmas were isolated from 21 tracheal swabs taken at necropsy. These mycoplasmas, together with six isolated from the equine respiratory tract by other workers, were subjected to biochemical and serological tests. Other properties examined in certain representative strains were appearance under the electron microscope, ability to adsorb or agglutinate the erythrocytes of various animal species and the electrophoretic pattern of the cell proteins. On the basis of these test, mycoplasmas from the equine respiratory tract were divided into seven species. Three species belonged to the genus Acholeplasma, members of which do not require sterol for growth, and were identified as A. laidlawii, A. oculi (formerly A. oculusi) originally isolated from the eyes of goats, and a recently named species A. equifoetale, previously isolated from aborted equine fetuses. Of the four sterol-dependent Mycoplasma species, one was indentified as M. pulmonis, a common rodent pathogen. Another cross-reacted serologically with M. felis and should probably be classified as that species. The other two species probably represent new species peculiar to the horse. One of these, represented by the strains N3 and N11, ferments glucose and is serologically distinct from 19 recognized species of glucose-utilizing mycoplasmas and from two species which do not metabolize either glucose or arginine. The other species, represented by four strains, hydrolyses arginine and, because it is serologically distinct from all the named arginine-hydrolysing Mycoplasma species, the name M. equirhinis sp.nov. is proposed for it. Of the seven species, only M. pulmonis and the glucose-utilizing species represented by N3 and N11 were found exclusively in horses with acute respiratory disease. A. oculi was isolated from an apparently normal horse. The other four species were found in normal horses as well as those with respiratory disease, although three out of the four strains of M. equirhinis were from sick horses.
PMCID: PMC2130589  PMID: 807616
24.  Occurrence of Ureaplasma parvum and Ureaplasma urealyticum in Women with Cervical Dysplasia in Katowice, Poland 
Journal of Korean Medical Science  2009;24(6):1177-1181.
The aim of this study was to evaluate the occurrence of genital mycoplasmas, especially Ureaplasma parvum and Ureaplasma urealyticum, in women with atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesions (LSIL) and high grade squamous intraepithelial lesions (HSIL), compared to women with normal cytology living in Katowice, Poland. Two sterile swabs were used to obtain material from the posterior vaginal fornix of 143 women with squamous intraepithelial lesions and 39 healthy women: first for general bacteriology, second for detection of urogenital mycoplasmas using Mycoplasma IST2 kit. From each positive Mycoplasma IST2 culture DNA was isolated and PCR was performed for identification of U. parvum and U. urealyticum. Mycoplasma IST was positive in 34.1% cases. Urogenital mycoplasmas were demonstrated in women with HSIL significantly more often compared to women with LSIL, ASCUS, and with normal cytology. DNA of U. parvum was demonstrated in majority of Mycoplasma IST2-positive cases, U. urealyticum DNA-only in 9 (4.9%). Predominance of 3/14 serovars of U. parvum was demonstrated. U. urealyticum biovar 2 was present more often in women with squamous intraepithelial lesions.
PMCID: PMC2775870  PMID: 19949678
Mycoplasma hominis; Ureaplasma urealyticum; ASCUS; LSIL; HSIL
25.  Mycoplasma species recovered from the reproductive tracts of western Canadian cows. 
Samples of cervico-vaginal mucus from 633 animals from 110 herds were cultured and yielded the following mycoplasmas: T-strain--88: Mycoplasma bovigenitalium--79, Mycoplasma spp. (Leach Group 7)--7, Acholeplasma laidlawii--4, Mycoplasma bovirhinis--2 and one not typable. Uterine exudates and endometrial scrapings from 80 infertile cows in two herds were examined. Four animals were positive, M. bovigenitalium was isolated three times, A. laidlawii and Mycoplasma arginini once each. Sixty-five normal uterine contents from pregnant cows were examined, one yielded M. bovigenigalium and the same organism was recovered from the fetal kidney. T-strain mycoplasma, M. bovigenitalium and other Mycoplasma spp. appear to be a part of the normal flora of the cervico-vaginal region of clinically normal one and two year old bred heifers in Alberta and Saskatchewan. Although M. arginini was not recovered from the cervico-vaginal region, a single recovery was made from the uterus of an infertile cow.
PMCID: PMC1277433  PMID: 1125831

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