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Background

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.

Methods

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.

Results

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.

Conclusion

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.

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%).

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.

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.

Cultures for mycoplasmatales, viruses and bacteria were made from bovine vulvar swabs to determine whether ureaplasma was associated with a clinical granular vulvitis observed in 16 Ontario dairy herds. Ureaplasma was isolated from 23.5% of 34 clinically normal cows, 74% of 27 cows with mild to moderate vulvar hyperemia but no discharge and 100% of 20 cows with acute vulvar hyperemia accompanied by purulent discharge. There were statistically significant differences in rates of isolation among clinical groups. Mycoplasma bovigenitalium was isolated from 7.7% and 20% of cows with moderate or acute vulvitis respectively but not from normal cows. Haemophilus somnus was isolated from 25% of cows with acute vulvitis. There were no significant differences in isolations of Escherichia coli, Corynebacterium pyogenes and alpha-hemolytic streptococcus between normal and clinically affected animals. Cultures of 135 repeat samples from 33 cows revealed that ureaplasma persisted in some animals for at least three months. No viruses were isolated from any of the animals in this study.

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.

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.

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.

One hundred and thirty-two penial-preputial swabbings, 140 raw and 42 processed semen samples were cultured for mycoplasmas. Mycoplasma or acholeplasma were recovered from 87, 32 and one respectively, while ureaplasmas were recovered from 46, 34 and six respectively.

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.

Urethral specimens from 726 patients with nongonococcal urethritis (NGU) were examined for Chlamydia trachomatis, Ureaplasma urealyticum, and Mycoplasma hominis. Chlamydiae were isolated from 35.9% of ureaplasma-positive patients and from 36.5% of ureaplasma-negative patients. Ureaplasmas were isolated from 52.5% of chlamydia-positive patients and from 53.1% of chlamydia-negative patients, an observation which contrasts with that of some workers who have suggested that ureaplasmas are significantly associated with chlamydia-negative NGU. Furthermore, the numbers of ureaplasmas isolated from patients who did or did not harbour chlamydiae were not significantly different nor was there a particular association of ureaplasmas with chlamydia-negative NGU in patients experiencing their first episode of disease. In addition, M. hominis was not isolated more frequently from those from whom chlamydiae were or were not isolated. The only significant associations were the isolation of M. hominis from patients who were ureaplasma-positive and of ureaplasmas from those who were M. hominis-positive. These findings do not necessarily mitigate against ureaplasmas being responsible for some cases of chlamydia-negative NGU.

Granular vulvitis was reproduced in ten virgin heifers following vulvar inoculation with strains of ureaplasma previously isolated from natural cases. The disease appeared one to three days postinoculation and was characterized by vulvar swabs but not from the upper mucopurulent discharge. At necropsy 13 to 41 days later, ureaplasmas were recovered consistently from vulvar swabs but not from the upper reproductive tract. It was concluded that some strains of ureaplasma are pathogenic and should be viewed as a cause of bovine granular vulvitis.

Two field isolates of Ureaplasma diversum spp. were used to infect heifers at the time of insemination in a preliminary study to observe the effect of infection on early pregnancy. M84-14c-1 was a field isolate from a bull's prepuce typed by immunofluorescence to be similar to U. diversum strain T-44 (Group C). M84-477c-4 was a field isolate from bovine semen typed by immunofluorescence to be similar to U. diversum strain T-288 (Group A). All three heifers infected with M84-477c-4 had a mild granular vulvitis at some time during the trial. None was pregnant when slaughtered 27 days after infection. The result of infection with M84-14c-1, a preputial isolate, was not consistent. One heifer had no infection and a normal pregnancy, one heifer was infected with an abnormal pregnancy, and one heifer was open but ureaplasmas were not detected until day 17 of the trial.

Objective

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.

Methods

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.

Results

(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.

Conclusion

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.

Ureaplasma urealyticum organisms (ureaplasmas) and Mycoplasma hominis organisms (mycoplasmas) were sought in mid-stream urines collected from 200 men and 200 women attending hospital with conditions of a non-venereal nature. In addition, the urines from 100 male and 100 female healthy volunteers were examined. Overall, ureaplasmas were isolated four times more often than mycoplasmas. In individuals less than 50 years of age, the organisms were found in about 20% of men and about 40% of women. In individuals 50 years or older, they were found about one-third to one-half as frequently. Centrifugation of urine and examination of the resuspended deposit did not increase the isolation rates. In men, the numbers of organisms in the urine were usually small (less than or equal to 10(3) c.c.u./ml) with less than tenfold more in the urine of women. The occurrence of 51- greater than 1000 leucocytes per mm3 in some of the urines was not associated with either the presence or an increased number of ureaplasmas/mycoplasmas, whereas they were associated with the presence of 10(5) or more bacteria/ml. The significance of these findings in the context of defining the role of ureaplasmas/mycoplasmas in genital-tract disease is discussed.

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.

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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.

The criteria that need to be fulfilled before regarding a mycoplasma as a cause of non-gonococcal urethritis (NGU) are outlined. Of the seven mycoplasmas that have been isolated from the human genitourinary tract, most cannot be considered as contenders for causing NGU. Although there is no evidence to support an etiological role for Mycoplasma hominis, it may be unwise to ignore this mycoplasma in view of its known pathogenicity in other situations. The cumulative weight of evidence indicates that strains of Ureaplasma urealyticum (ureaplasmas) cause NGU in some patients. The reason for their occurrence in the urethra of some men without disease needs to be established. Ureaplasmas do not seem to cause post-gonococcal urethritis. The role in NGU of M. genitalium, newly discovered in the male urethra, is unknown, but its biological features, morphological appearance, and ability to cause genital disease in marmosets suggest that it may be pathogenic for man.

The cervicovaginal and endometrial isolation rates of Ureaplasma urealyticum and Mycoplasma hominis and relevant demographic data were obtained at the time of laparoscopy in 193 women from infertile marriage. For comparative purposes, fertile women undergoing laparoscopy for tubal ligation (n = 56) or other purposes (n = 64) were also cultured. Blacks were more likely than caucasians to be infected with either organism in all population types (p less than or equal to .05); however, no differences were noted in cervicovaginal carriage rates for blacks in different patient populations. M. hominis was isolated more frequently from tubal reanastomosis patients and less often from infertile patients, p less than or equal to .001. No differences were noted among the infertile subpopulations. Although the isolation rate of U. urealyticum from the different patient populations was similar, one subpopulation within the infertile population (male factor) was identified in which the prevalence of ureaplasmal infection of the female's lower genital tract was over twice as high (p less than or equal to .005) as in other infertile women. Yet there were no statistically significant differences in the demographic data of this subpopulation as compared to the population of infertile women as a whole. No other clinical subpopulation with single or multiple diagnoses not including male factor had an increased prevalence of infection. Eighty percent of infected, infertile couples had no clinical evidence of male factor infertility, indicating that only certain individuals are affected.This possibly explains why previous studies involving small numbers of patients without regard to clinical subpopulations have failed to show significant differences between infected and uninfected couples.(ABSTRACT TRUNCATED AT 250 WORDS)

Duplicate vaginal swabs were collected from 100 women, and comparisons were made between an in-house broth-agar culture system and a commercially available kit, the Mycoplasma IST kit (bioMérieux), for the detection of Mycoplasma hominis and Ureaplasma urealyticum. There was good agreement between the two systems for detection of the genital mycoplasmas in terms of sensitivity, with values of > 92% being obtained. In terms of specificity, the mutual comparisons were less favorable, though specificity values of > 72% were obtained. Statistically there was no significant difference in the performance of the two tests (P < 0.1 for both M. hominis and U. urealyticum). While the broth-agar culture system was considerably less expensive than the kit, the Mycoplasma IST kit provided additional information on antibiotic susceptibilities and had the advantages of a shelf life of up to 12 months and not requiring the preparation of culture media. The prevalences of colonization obtained for M. hominis and U. urealyticum were extremely high in this randomly selected group of women from periurban and rural settlements in the Eastern Highlands of Papua New Guinea, being > or = 70% for M. hominis and > or = 78% for U. urealyticum. colonization with both genital mycoplasmas simultaneously was also very common, with > or = 60% of women being colonized by both M. hominis and U. urealyticum.

Vaginal specimens were collected two to three times a week for 1 month from seven nurses. A total of 65 specimens were collected. Each sample consisted of three swabs and a saline wash. Semiquantitation of anaerobic and aerobic bacteria, mycoplasma and ureaplasma, and yeast was performed. Numerous species were recovered in each specimen; at least 37 species were isolated. Lactobacilli, Corynebacterium, Ureaplasma, Mycoplasma, Bacteroides melaninogenicus, and Candida albicans, when present, tended to remain throughout the entire month. Other organisms were present on a more sporadic basis. The number of organisms varied greatly during the sampling for each individual, whereas the types of organisms isolated from a particular subject remained relatively constant.

The results of a double-blind therapeutic trial on 217 men with nongonococcal urethritis (NGU) show that minocycline was more effective than rifampicin. Before treatment Chlamydia trachomatis was isolated from 43% of men, Ureaplasma urealyticum from 59%, and Mycoplasma hominis from 22%. Chlamydiae and ureaplasmas were isolated less frequently from men with a recent history of NGU. Minocycline was given to 94 patients, and after treatment chlamydiae were isolated from only one of 40 initially chlamydia-positive patients and ureaplasmas from only five of 57 initially ureaplasma-positive patients. Although most patients responded clinically, failure and partial recovery rather than complete recovery were observed more often among those who were infected with ureaplasmas. Rifampicin was given to 123 patients, after which chlamydiae were isolated from only one of 53 initially chlamydia-positive men whereas ureaplasmas, insensitive to the antibiotic in vitro, were isolated from 55 of 68 men who had initially positive results. Patients infected with ureaplasmas failed to respond to rifampicin treatment significantly more often than those who were not infected. This was also observed when only patients who had never had NGU or who had not had a recent episode were considered. Furthermore, 24 (44%) of the 55 men whose ureaplasmas persisted failed to recover whereas only one (7·7%) of 13 men whose ureaplasmas disappeared did not respond to treatment. These results suggested that ureaplasmas were a cause of urethritis in some of the men (an estimated 10% at least). In addition, Reiter's disease developed in two men treated with rifampicin from whom only ureaplasmas had been isolated initially. M. hominis did not seem to have an important pathogenic role in NGU and there was evidence that ureaplasmas were an unlikely cause of urethritis in some men since the organisms persisted despite complete clinical recovery.

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.

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.

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.