Evaluation of commercial, customized microdilution plates for Ureaplasma parvum, Ureaplasma urealyticum, and Mycoplasma hominis antimicrobial susceptibility testing and determination of antimicrobial resistance prevalence in France.

Antimicrobial susceptibility testing (AST) of human mycoplasmas using microdilution is time-consuming. In this study, we compared the performance of MICRONAUT-S plates (Biocentric-Bruker) designed for AST of Ureaplasma parvum, Ureaplasma urealyticum, and Mycoplasma hominis with the results using the Clinical & Laboratory Standards Institute (CLSI) reference method. Then, we investigated the prevalence and mechanisms of resistance to tetracyclines, fluoroquinolones, and macrolides in France in 2020 and 2021. The two methods were compared using 60 strains. For the resistance prevalence study, U. parvum-, U. urealyticum-, and M. hominis-positive clinical specimens were collected for 1 month each year in 22 French diagnostic laboratories. MICs were determined using the MICRONAUT-S plates. The tet(M) gene was screened using PCR, and fluoroquinolone resistance-associated mutations were screened using PCR and Sanger sequencing. Comparing the methods, 99.5% (679/680) MICs obtained using the MICRONAUT-S plates concurred with those obtained using the CLSI reference method. For 90 M. hominis isolates, the tetracycline, levofloxacin, and moxifloxacin resistance rates were 11.1%, 2.2%, and 2.2%, respectively, with no clindamycin resistance. For 248 U. parvum isolates, the levofloxacin and moxifloxacin resistance rates were 5.2% and 0.8%, respectively; they were 2.9% and 1.5% in 68 U. urealyticum isolates. Tetracycline resistance in U. urealyticum (11.8%) was significantly (P < 0.001) higher than in U. parvum (1.2%). No macrolide resistance was observed. Overall, the customized MICRONAUT-S plates are a reliable, convenient tool for AST of human mycoplasmas. Tetracycline and fluoroquinolone resistance remain limited in France. However, the prevalence of levofloxacin and moxifloxacin resistance has increased significantly in Ureaplasma spp. from 2010 to 2015 and requires monitoring. IMPORTANCE: Antimicrobial susceptibility testing of human urogenital mycoplasmas using the CLSI reference broth microdilution method is time-consuming and requires the laborious preparation of antimicrobial stock solutions. Here, we validated the use of reliable, convenient plates designed for antimicrobial susceptibility testing that allows the simultaneous determination of the MICs of eight antibiotics of interest. We then investigated the prevalence and mechanisms of resistance of each of these bacteria to tetracyclines, fluoroquinolones, and macrolides in France in 2020 and 2021. We showed that the prevalence of levofloxacin and moxifloxacin resistance has increased significantly in Ureaplasma spp. from 2010 to 2015 and requires ongoing monitoring.

Membrane inflammasome activation by choriodecidual Ureaplasma parvum infection without intra-amniotic infection in a Non-Human Primate model†.

Intrauterine infection is a significant cause of neonatal morbidity and mortality. Ureaplasma parvum is a microorganism commonly isolated from cases of preterm birth and preterm premature rupture of membranes (pPROM). However, the mechanisms of early stage ascending reproductive tract infection remain poorly understood. To examine inflammation in fetal (chorioamnionic) membranes we utilized a non-human primate (NHP) model of choriodecidual U. parvum infection. Eight chronically catheterized pregnant rhesus macaques underwent maternal-fetal catheterization surgery at ~105-112 days gestation and choriodecidual inoculation with U. parvum (105 CFU/mL, n =4) or sterile media (controls; n = 4) starting at 115-119 days, repeated at 5-day intervals until C-section at 136-140 days (term=167 days). The average inoculation to delivery interval was 21 days, and Ureaplasma infection of the amniotic fluid (AF) was undetectable in all animals. Choriodecidual Ureaplasma infection resulted in increased fetal membrane expression of MMP-9 and PTGS2, but did not result in preterm labor or increased concentrations of AF pro-inflammatory cytokines. However, membrane expression of inflammasome sensors, NLRP3, NLRC4, AIM2, and NOD2, and adaptor ASC (PYCARD) gene expression were significantly increased. Gene expression of IL-1ß, IL-18, IL-18R1  , CASPASE-1, and pro-CASPASE-1 protein increased with Ureaplasma infection. Downstream inflammatory genes MYD88 and NFκB (Nuclear factor kappa-light-chain-enhancer of activated B cells) were also significantly upregulated. These results demonstrate that choriodecidual Ureaplasma infection, can cause activation of inflammasome complexes and pathways associated with pPROM and preterm labor prior to microbes being detectable in the AF.

Analysis of gastric fluid in preterm newborns supports the view that the amniotic cavity is sterile before the onset of parturition: a retrospective cohort study.

OBJECTIVES: To compare the frequency of Ureaplasma-positive gastric fluid (GF) cultures based on the cause and mode of delivery in preterm newborns. METHODS: This retrospective cohort study included women with a singleton pregnancy who delivered prematurely (between 23+0 and 32+0 weeks of gestation, n=464) at a single university hospital in South Korea. The newborns' GF was obtained on the day of birth via nasogastric intubation. The frequency of Ureaplasma spp. in GF cultures was measured and compared according to the cause and mode of delivery. RESULTS: Ureaplasma spp. was detected in 20.3 % of the GF samples. The presence of Ureaplasma spp. was significantly higher in the spontaneous preterm birth group than in the indicated preterm birth group (30.2 vs. 3.0 %; p<0.001). Additionally, Ureaplasma spp. was more frequently found in the vaginal delivery group than in the cesarean delivery group, irrespective of the cause of preterm delivery [indicated preterm birth group (22.2 vs. 1.9 %, p=0.023); spontaneous preterm birth group (37.7 vs. 24.2 %, p=0.015)]. CONCLUSIONS: Ureaplasma spp. were found in 20.3 % of the GFs. However, only 1.9 % of newborns in the indicated preterm birth group with cesarean delivery had a Ureaplasma-positive GF culture.

The Relationship Between Ureaplasma Species and Male Infertility: Pathogenicity, Biology, Diagnosis, and Treatment.

Objective: Ureaplasma spp. comprise a group of mycoplasmas containing two human-associated species, namely, Ureaplasma urealyticum (UUR) and Ureaplasma parvum (UPA). The characterization of Ureaplasma species as pathogens contributing to male infertility remains a subject of considerable controversy. While numerous authors have proposed a relationship between UUR and changes in fertility, there is limited evidence supporting the involvement of UPA in this context. There has been an increased focus on Ureaplasma spp. and its potential role in the development of male infertility, especially over the past few years. The review aims to clarify the relationship between Ureaplasma species and male infertility. Methods: Firstly, we introduce a background of the appropriate biology including growth characteristics, the divided biovars, and the transmission pathways. Secondly, we examine the studies that support a causal role for Ureaplasma spp. in the development of infertility in the last 30 years. Finally, the diagnosed method, antimicrobial susceptibility, and potential therapeutic considerations are evaluated. Results: UPA and UUR can impair semen motility. The species of Ureaplasma spp., the sexual history of the patient, the number of sexual partners, the load of Ureaplasma, and antimicrobial resistance are expected to constitute key risk factors in the development of male infertility. In terms of treatment, Doxycycline remains the drug of first choice for ureaplasmal infections. Conclusion: Ureaplasma spp. are not simply "innocent bystanders" in infertility and may indeed be an "underestimated enemy of human reproduction". Ureaplasma spp. can be considered an etiological agent in unexplained infertility and a useful marker.

Does vaginal bacterial colonization contribute to preterm birth in women with asymptomatic shortened cervix?

PURPOSE: The aim of this study is to describe the typical microbial spectrum and the influence of distinct vaginal infections on preterm birth in pregnancies affected by cervical incompetence. METHODS: 327 patients were admitted because of asymptomatic shortening of the cervix in the second and third trimester of pregnancy. Clinical data such as age, cervical length, gestational age at admission and at delivery and vaginal microbiologic findings were collected and analyzed. RESULTS: The spectrum of germs in the vagina revealed seven different distinct species; the most common bacteria were Ureaplasma spp. and E. coli. In 327 included patients, 217 revealed a bacterial colonization, 110 did not. Most common bacteria in women with preterm birth before 34 weeks were Ureaplasma spp., while E. coli was most common in women undergoing preterm birth after 34 weeks. Nevertheless, the rates of occurrence of these bacterial taxa were not significantly different between who underwent preterm birth to those who did not. CONCLUSIONS: This study gives an overview over the vaginal bacterial colonization in pregnant women with cervical incompetence. The clinical relevance of vaginal bacterial colonization remains unclear.

Antimicrobial Activity of N,N-Diethyldithiocarbamate against Ureaplasma parvum and Ureaplasma urealyticum.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts, although their overgrowth can lead to infection in the urogenital tract and at distal sites. Furthermore, ureaplasmas lack a cell wall and do not synthesize folic acid, which causes all ß-lactam and glycopeptide antibiotics, and sulfonamides and diaminopyrimidines, to be of no value. The antibiotics used in therapy belong to the fluoroquinolone, tetracycline, chloramphenicol and macrolide classes. However, the growing incidence of antibiotic-resistant Ureaplasma spp. in the population becomes a problem. Thus, there is a need to search for new drugs effective against these bacteria. Since 1951, the FDA-approved, well-tolerated, inexpensive, orally administered drug disulfiram (DSF) has been used in the treatment of chronic alcoholism, but recently, its antimicrobial effects have been demonstrated. The main biological metabolite of DSF, i.e., N,N-diethyldithiocarbamate (DDC), is generally believed to be responsible for most of the observed pharmacological effects of DSF. In the presented studies, the effect of DDC at concentrations of 2 µg/mL, 20 µg/mL and 200 µg/mL on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested for the first time. The results indicated that all the used DDC concentrations showed both bacteriostatic and bactericidal activity against both tested strains.

Effects of Ureaplasma parvum infection in the exosome biogenesis-related proteins in ectocervical epithelial cells.

Ureaplasma parvum is a mycoplasma commonly associated with female reproductive pathologies, such as preterm birth and infertility. It can survive intracellularly and utilize exosomes to propagate infection and its virulence factors. This study explored the differential protein composition of exosomes derived from normal and U. parvum-infected cells. We also investigated the impact of U. parvum on exosome biogenesis in ectocervical epithelial cells. Ectocervical epithelial (ECTO) cells were infected with U. parvum, and immunocytochemical staining was performed using U. parvum-specific marker multiple banded antigen (mba) and exosome marker CD9. NanoLC-MS/MS analysis was conducted to identify differentially expressed proteins in exosomes. Ingenuity Pathway Analysis (IPA) was performed to identify affected canonical pathways and biological functions associated with the protein cargo of exosomes. Western blot analysis of ECTO cells validated the proteomic findings in ECTO cells. U. parvum exhibited colonization of ECTO cells and colocalization with CD9-positive intraluminal vesicles. Proteomic analysis revealed decreased protein abundance and distinct protein profiles in exosomes derived from U. parvum-infected ECTO cells. Differentially expressed proteins were associated with clathrin-mediated endocytosis and various signaling pathways indicative of infection, inflammation, and cell death processes. Additionally, U. parvum infection altered proteins involved in exosome biogenesis. In ECTO cells, U. parvum infection significantly decreased clathrin, ALIX, CD9, and CD63 and significantly increased TSG101, Rab5, Rab35, and UGCG. These findings contribute to our understanding of the infection mechanism and shed light on the importance of exosome-mediated communication in the pathophysiology of diseases affecting the cervix, such as cervicitis and preterm birth.

The Role of Ureaplasma and Mycoplasma Species in Recurrent Lower Urinary Tract Infection in Females: A Scoping Review.

It is unknown if recurrent urinary tract infection in the gynecologic population is associated with Mycoplasma and Ureaplasma genitourinary infections. The purpose of this scoping review is to highlight the literature surrounding Mycoplasma and Ureaplasma infections in the setting of recurrent urinary tract infections in the gynecologic population. MEDLINE ALL and Embase were searched to retrieve articles published in or after 1950 through 2024. Studies included were those with adults over age 18, non-pregnant, diagnosed with recurrent urinary tract infection and concurrent genitourinary infection with Ureaplasma or Mycoplasma published in English. Study designs eligible were quantitative, qualitative, and mixed methods studies. Publication types were also extended to conference abstracts and unpublished data. 2 independent investigators systematically performed title/abstract screening and full-text review using standardized inclusion criteria. For disagreements in either title and abstracts or full-text articles, consensus was reached through discussion by the 2 screeners and/or a 3rd final adjudicator. Screening and data extraction were performed on Covidence, a web-based platform for systematic review management. There were 1170 studies identified before title and abstract screening. 26 full-text articles were reviewed for eligibility. Of these, 23 full-text studies were excluded. 3 studies met full inclusion criteria and data extraction was performed on these 3 studies. There were 2 additional studies included after identification via other methods. There is a need for more recent and robust studies examining the role of Ureaplasma and Mycoplasma genitourinary infections amongst gynecologic patients with recurrent urinary tract infections.

Mycoplasma hominis increases the risk for Ureaplasma parvum infection in Human immunodeficiency virus infected pregnant women.

INTRODUCTION: Mycoplasma hominis and Ureaplasma parvum have been recently linked to sexually transmitted diseases and other conditions. There are a limited number of studies conducted on South African pregnant women that have assessed the prevalence and risk factors for genital mycoplasmas. METHODOLOGY: This study included 264 HIV infected pregnant women attending the King Edward VIII antenatal clinic in eThekwini, South Africa. DNA was extracted using the PureLink Microbiome kit and pathogens were detected using the TaqMan Real-time PCR assays. The statistical data analysis was conducted in a freely available Statistical Computing Environment, R software, version 3.6.3 using the RStudio platform. RESULTS: The prevalence of M. hominis and U. parvum, was 215/264 (81.4%), and 203/264 (76.9%), respectively. In the M. hominis positive group, a significantly (p = 0.004) higher proportion, 80.5% tested positive for U. parvum infection when compared to 61.2% among the M. hominis negative. Of the U. parvum positive women, a significantly (p = 0.004) higher proportion of women (85.2%) tested positive for M. hominis when compared to 68.9% among the U. parvum negative. In the unadjusted and adjusted analysis, being M. hominis positive increased the risk for U. parvum by approximately 3 times more (p = 0.014) and 4-fold (p = 0.008), respectively. CONCLUSIONS: This study showed a significant link between M. hominis and U. parvum infection. To date, there are a limited number of studies that have investigated M. hominisbeing a risk factor for U. parvum infection. Therefore, the data presented in the current study now fills in this gap in the literature.

Estimates of the incidence, prevalence, and factors associated with common sexually transmitted infections among Lebanese women.

BACKGROUND: We analyzed the prevalence of active infection with common curable sexually transmitted infections (STIs) including N. gonorrhea, C. trachomatis, T. vaginalis, and T. pallidum, as well as active infection with HPV, herpes simplex virus types I (HSV-1) and II (HSV-2), M. hominis, M. genitalium, C. albicans, and Ureaplasma in 351 Lebanese women. METHODS: A cross-sectional study, involving 351 sexually active women, 40 years or younger, who were recruited from outpatient Obstetrics and Gynecology clinic attendees between September 2016 and November 2017. RESULTS: The prevalence of active infection was low at 0.3% for N. gonorrhea, 0.6% for HSV-2, 2.8% for C. trachomatis, and 2.9% for any curable STIs. Prevalence of active HPV infection was high assessed at 15.7% for high-risk and 12.2% for low-risk genotypes. Furthermore, the prevalence was 2.0% for M. genitalium, 6.8% for ureaplasma, 13.7% for Candida albicans, and 20.5% for M. hominis. No active infections with T. vaginalis, T. pallidum, or HSV-1 were observed. Significant age differences were noted in the prevalence of high-risk and low-risk HPV genotypes, but no such differences were noted in the prevalence of other infections. No appreciable variations were identified in the prevalence of key STIs based on smoking, marital status, or the number of sexual partners. CONCLUSIONS: The study documented active infection with substantial prevalence for multiple STIs among women attending outpatient gynecology and obstetrics clinics in Lebanon. These findings underscore the importance of strengthening STI surveillance, linkage to care, and prevention interventions in reducing STI incidence among women.

Clinical and uterine cervix characteristics of women with Mycoplasma and Ureaplasma in genital discharge.

OBJECTIVE: The objective of this study was to assess the clinical and uterine cervix characteristics of patients displaying vaginal discharge with positive results for Mycoplasma sp. and/or Ureaplasma spp. METHODS: An analytical cross-sectional study involving women aged 18-45 years was conducted. Microbiological assessments included Ureaplasma and Mycoplasma cultures, as well as human papillomavirus hybrid capture using ecto and endocervix swabs. All tests were two-tailed, and significance was set at p<0.05. RESULTS: Among 324 women, Ureaplasma prevalence was 17.9%, and Mycoplasma prevalence was 3.1%. The Ureaplasma-positive group exhibited a higher frequency of urinary tract infections (39.1 vs. 19%, p=0.002) and human papillomavirus (39.7 vs. 12.8%, p≤0.001) compared with controls. The Mycoplasma-positive group showed a higher frequency of non-contraceptive use compared with controls (66.2 vs. 30.0%, p=0.036). Abnormal colposcopic findings were more prevalent in the Mycoplasma/Ureaplasma-positive group than in controls (positive: 65% vs. control: 35%, p=0.001). Pap smear findings did not differ between the groups. CONCLUSION: Ureaplasma spp. was associated with urinary tract infections and human papillomavirus, while the presence of Mycoplasma sp. was linked to reduced contraceptive use. When analyzing both pathogens together, a higher frequency of abnormal colposcopic findings was observed, with no difference in cytological findings in the positive group.

Prenatal exposure to intra-amniotic infection with Ureaplasma species increases the prevalence of bronchopulmonary dysplasia.

OBJECTIVES: The present study investigated the relationship between bronchopulmonary dysplasia (BPD) and intra-amniotic infection with Ureaplasma species. METHODS: This was a single-center, retrospective cohort study. Patients with singleton pregnancies who underwent inpatient management at our department for preterm premature rupture of membranes (PPROM), preterm labor, cervical insufficiency, and asymptomatic cervical shortening at 22-33 gestational weeks were included. Amniocentesis was indicated for patients with PPROM or an elevated maternal C-reactive protein level (≥0.58 mg/dL). Patients with an amniotic fluid IL-6 concentration ≥3.0 ng/mL were diagnosed with intra-amniotic inflammation, while those with positive aerobic, anaerobic, M. hominis, and Ureaplasma spp. cultures were diagnosed with microbial invasion of the amniotic cavity (MIAC). Patients who tested positive for both intra-amniotic inflammation and MIAC were considered to have intra-amniotic infection. An umbilical vein blood IL-6 concentration >11.0 pg/mL indicated fetal inflammatory response syndrome (FIRS). The maternal inflammatory response (MIR) and fetal inflammatory response (FIR) were staged using the Amsterdam Placental Workshop Group Consensus Statement. RESULTS: Intra-amniotic infection with Ureaplasma spp. was diagnosed in 37 patients, intra-amniotic infection without Ureaplasma spp. in 28, intra-amniotic inflammation without MIAC in 58, and preterm birth without MIR/FIR and FIRS in 86 as controls. Following an adjustment for gestational age at birth, the risk of BPD was increased in patients with intra-amniotic infection with Ureaplasma spp. (adjusted odds ratio: 10.5; 95% confidence interval: 1.55-71.2), but not in those with intra-amniotic infection without Ureaplasma spp. or intra-amniotic inflammation without MIAC. CONCLUSION: BPD was only associated with intra-amniotic infection with Ureaplasma species.

Immunocompromised teenager with meningitis caused by Ureaplasma parvum.

Infection in the immunocompromised patient is often challenging on multiple levels. It can be difficult to distinguish between manifestations of the underlying disease, infection or malignancy. Symptoms may be vague or even absent, deviations in the common inflammatory parameters discrete, imaging findings scarce and the causative microbe may be a true pathogen as well as opportunistic. Here, we report an immunosuppressed female in her late teens with a purulent meningitis due to Ureaplasma parvum-a very rare cause of infection in the central nervous system of adults. We wish to highlight the relevance of intracellular pathogens and the need to actively search for these microbes, especially when response to broad-spectrum antibiotic treatment is absent. Furthermore, we emphasise the need for adequate molecular microbial diagnostics in search of microbes that are difficult to identify by culture and where serology and antigen tests may be absent or unreliable due to immune suppression.

Occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens

Mycoplasmaspp. and Ureaplasmaspp. belong tohumans'genitourinary microbiota and sometimesare associated with infections of the genitourinarytract. The aim of this study was to evaluate the occurrence of Mycoplasmaspp. and Ureaplasmaspp. in genital specimens from patients of the 15thRegional de Saúde of ParanáState, Brazil, and to correlate the results with clinical and laboratory data.A retrospective cross-sectional study was conducted,based on the analysis of results of vaginal, endocervical, urine andurethral culture for mycoplasmas from patients attended in areference laboratory, from January 2009 to December 2016. We evaluated 2,475 results of culture for mycoplasmas. A total of 50.8% patients were positive for mycoplasmas. Of these, 76.8%had positive culture exclusively for Ureaplasmaspp. and 4.7% for Mycoplasmahominis. Both microorganisms were isolated in the microbiology culture of 18.5% of patients. Among the positive culture, 81.4% had significant concentrations.Bacterialvaginosis was the most common alteration observed in association with mycoplasmas.Thehigh positivity of cultures for mycoplasmas, especially Ureaplasmaspp. found in our study, highlightthe presence of these microorganisms in many of the genital tract disorders that can be sexually transmitted and, consequently, should not be neglected.

Micoplasma hominis y Ureaplasma spp en mujeres que consultan por infertilidad / Mycoplasma Hominis and Ureaplasma Spp. in women attending to infertility consultations

RESUMEN Introducción: Las infecciones por micoplasmas y ureaplasmas pueden producir fallos en la reproducción y vincularse con problemas de infertilidad femenina. Objetivo: Determinar la frecuencia de infecciones por Micoplasma hominis y Ureaplasma spp en mujeres que consultan por infertilidad e identificar si existe asociación entre las infecciones detectadas y los antecedentes de infecciones de transmisión sexual y enfermedad inflamatoria pélvica, procederes ginecológicos y síntomas de infecciones. Métodos: Se realizó un estudio descriptivo transversal, para evaluar muestras de exudados endocervicales de 175 mujeres, con edades entre 20 y 45 años, provenientes de la consulta de infertilidad del Instituto Nacional de Endocrinología, entre junio de 2016 y enero de 2017. Para la detección de micoplasmas urogenitales se utilizó el juego de reactivos Myco Well D-One. Se tuvieron en cuenta los aspectos éticos y se utilizó la prueba Chi Cuadrado para evaluar la significación estadística de las posibles asociaciones. Resultados: De las 175 muestras evaluadas, 102 (58,1 por ciento) mostraron la presencia de infecciones, de ellas 65 correspondieron a Ureaplasma spp (37,1 por ciento), 11 a Micoplasma hominis (6,2 por ciento), y 26 a asociaciones de Micoplasma hominis y Ureaplasma spp (14,8 por ciento). Se identificó asociación entre las infecciones detectadas y la presencia de antecedentes de infecciones de transmisión sexual y enfermedad inflamatoria pélvica, no así con relación a los procederes ginecológicos y síntomas de infecciones. Conclusiones: La frecuencia total de infecciones fue relativamente alta y la especie más frecuente el Ureaplasma spp. Las infecciones detectadas estuvieron asociadas a algunos de los factores estudiados(AU)

ABSTRACT Introduction: Infections caused by Mycoplasmas and Ureaplasmas may result in faults in the reproduction process and can be linked to female infertility. Objective: To determine the frequency of infection by Mycoplasma hominis and Ureaplasma spp. in women who attend to infertility consultations and if these are associated with a history of sexually transmitted infections and pelvic inflammatory disease, gynaecological procedures and symptoms of infections. Methods: A descriptive cross-sectional study was conducted to evaluate samples of endocervical swabs of 175 women between the ages of 20 to 45 years, from the Infertility consultation of the National Institute of Endocrinology, during June 2016 to January 2017. For the detection of urogenital mycoplasmas it was used the reagents kit Myco Well D-One. There were taken into account the ethical aspects and it was used the chi-square test to assess the statistical significance of the possible associations. Results: Of the 175 evaluated samples, 102 (58.1 percent) showed the presence of infections, 65 of them corresponded to Ureaplasma spp (37.1 percent), 11 to Mycoplasma hominis (6.2 percent), and 26 associations of Mycoplasma hominis and Ureaplasma spp (14.8 percent). It was identified association between the detected infections and the presence of a history of sexually transmitted infections and pelvic inflammatory disease, but not with the gynaecological procedures and the symptoms of infections. Conclusions: The total frequency of infection was relatively high and the most prevalent specie was the Ureaplasma spp. The detected infections were associated with some of the factors studied(AU)

Identificación de especies de Mycoplasma y de Ureaplasma diversum en rodeos lecheros de Argentina / Identification of species of Mycoplasma and Ureaplasma diversum from Argentinian dairy herds

Varias especies de Mycoplasma y Ureaplasma diversum pueden causar enfermedades en el ganado bovino lechero, asociadas o no a manifestaciones clínicas. En nuestro país, ha sido detectada la presencia de solo tres especies de este grupo hasta el momento: Mycoplasma bovis, Mycoplasma californicum y Mycoplasma canadense. El objetivo del presente trabajo fue identificar otras especies de la familia Mycoplasmataceae. Se estudiaron treinta y cinco aislamientos compatibles con Mycoplasma spp. obtenidos a partir de diferentes muestras de bovinos, con o sin sintomatología clínica, provenientes de ocho rodeos ubicados en las provincias de Santa Fe, Córdoba, Buenos Aires y San Luis. Mediante el uso de reacciones en cadena de la polimerasa (PCR) específicas de especie se identificaron Mycoplasma bovigenitalum, Mycoplasma alkalescens, Mycoplasma bovirhinis y U. diversum, y mediante la amplificación y posterior secuenciación del espacio intergénico 16-23S ARNr se identificaron Mycoplasma arginini y M. californicum. La identificación de estas especies por primera vez en nuestro país es un hecho de Argentina relevancia, que representa un importante avance en el conocimiento para incluir estos patógenos en el diagnóstico diferencial de determinadas entidades clínico-patológicas de los bovinos de Argentina.

Several species of Mycoplasma and Ureaplasma diversum can cause diseases in dairy cattle, which can be associated or not with clinical manifestations. In our country, the presence of Mycoplasma bovis, Mycoplasma californicum and Mycoplasma canadense has been detected, being the only mycoplasma species identified so far. The objective of this study was to identify other species of the Mycoplasmataceae family. Thirty-five Mycoplasma spp.-like isolates obtained from different samples from cattle, with or without clinical symptoms, from eight herds located in the provinces of Santa Fe, Cordoba, Buenos Aires and San Luis were utilized in the present study. Through the use of species-specific polymerase chain reactions (PCR) Mycoplasma bovigenitalium, Mycoplasma alkalescens, Mycoplasma bovirhinis and U. diversum were identified and through amplification and further sequencing of the 16-23S rRNA intergenic spacer regions, Mycoplasma arginine and M. californicum were identified. The identification of these species represents an important advance in knowledge in order to include these pathogens in the differential diagnosis of certain clinical and pathological entities of cattle from Argentina.

Molecular characterization of ureaplasmas isolated from reproductive tract of goats and sheep from Brazil

Abstract Ovine/caprine ureaplasmas have not yet been assigned a species designation, but they have been classified into nine serotypes. Herein ureaplasmas were searched for in 120 samples of vulvo vaginal mucous from sheep and 98 samples from goats at 17 farms. In addition, semen samples were collected from 11 sheep and 23 goats. The recovered ureaplasma were from sheep and goats from animals without any reproductive disorder symptoms, but not all animals presented positive cultures. In sheep, 17 (68%) cultures of vulvovaginal mucous were positive for ureaplasma and 11 (27%) samples of semen presented positive cultures in animals with clinical signs of orchitis, balanoposthitis or low sperm motility. In goats four ureaplasma isolates were obtained from vulvovaginal mucus, but the semen samples were all negative. The isolates were submitted to Pulsed-field gel electrophoresis methodology and their 16S rRNA genes were sequenced. Fifty percent of ureaplasma recovered from sheep allowed for PFGE typing. Eleven isolates showed eight profiles genetically close to the bovine ureaplasmas. The 16S rRNA gene sequencing showed differences or similarities of isolates from sheep and goats, and the reference strains of bovine and human ureaplasma. Four clinical isolates from sheep were grouped separately. The studied ureaplasma isolates showed to be a diverse group of mollicutes.

Assessment of cow and farm level risk factors associated with Ureaplasma diversum in pasture-based dairy systems - A field study

ABSTRACT Potential risk factors for Ureaplasma diversum in the vaginal mucus of 1,238 dairy cows were included in a multivariate logistic regression model, based on the cow level (i.e., granular vulvovaginitis [+GVV], yearly milk production [4500 kg or more], pregnancy, predominance of Bos taurus [+Bos Taurus], score of corporal condition [at least 2.5], concomitant positivity for Escherichia coli [+E.coli]), and farm level i.e., milking room hygiene (-Milking room), dunghill location, and replacement female). Ureaplasma diversum was present in 41.1% of the samples. Independent risk factors for U. diversum were +GVV (odds ratio [OR], 1.31); +Mycoplasma spp (OR, 5.67); yearly milk production (4500 kg or more) (OR, 1.99); +Bos taurus (OR, 1.68); +E. coli (OR, 4.96); -milking room (OR, 2.31); and replacement females (OR, 1.89). Ureaplasma diversum vaginal colonization was strongly associated with Mycoplasma spp., E. coli, and number of pregnant cows.

Utilidad de la reacción de polimerasa en cadena convencional para la detección de Mycoplasma hominis, Ureaplasma spp. y Trichomonas vaginalis en muestras genitales de mujeres en consulta ambulatoria / Usefulness of conventional polymerase chain reaction for the detection of Mycoplasma hominis, Ureaplasma spp. and Trichomonas vaginalis in female outpatient's genital samples

Introduction: Trichomonas vaginalis, Mycoplasma hominis and Ureaplasma spp. are microorganisms responsible for genitourinary and pregnancy pathologies. Nucleic acid amplification methods have shown several advantages, but have not been widely studied for the detection of these microorganisms. Aim: To implement a conventional polymerase chain reaction (PCR) for the detection of the microorganisms and to compare its results versus the methods currently used at our laboratory. Material and Methods: 91 available samples were processed by PCR, culture (M. hominis y Ureaplasma spp.) and wet mount (T vaginalis). Results were compared and statistically analyzed by kappa agreement test. Results: 85, 80 and 87 samples resulted in agreement for the detection of M. hominis, Ureaplasma spp. y T. vaginalis, respectively. For M. hominis and Ureaplasma spp., agreement was substantial, whereas for T. vaginalis it was moderate, however, for the latter, PCR detected more cases than wet mount. Conclusion: We recommend the implementation of PCR for detection of T. vaginalis whereas culture kit is still a useful method for the other microorganisms.

Introducción: Trichomonas vaginalis, Mycoplasma hominis y Ureaplasma spp. son microorganismos causantes de patología genito-urinaria y durante el embarazo. Los métodos de amplificación de ácidos nucleicos han demostrado numerosas ventajas, pero no han sido ampliamente estudiados para la detección de estos microorganismos. Objetivo: Implementar una reacción de polimerasa en cadena convencional (RPC) para su detección y comparar sus resultados con los métodos actuales de nuestro laboratorio. Material y Métodos: Se procesaron 91 muestras mediante RPC, cultivo (M. hominis y Ureaplasma spp.) y observación microscópica al fresco (T. vaginalis). Los resultados fueron comparados y analizados estadísticamente mediante el test de concordancia kappa. Resultados: 85, 80 y 87 muestras tuvieron resultados concordantes para la detección de M. hominis, Ureaplasma spp. y T. vaginalis, respectivamente. Para M. hominis y Ureaplasma spp. el nivel de concordancia fue considerable mientras que para T. vaginalis fue moderado; sin embargo, para esta última, la RPC detectó más casos que la microscopia al fresco. Conclusión: Se recomienda la implementación de la RPC para la detección de T. vaginalis. Para M. hominis y Ureaplasma spp. el kit de cultivo continúa siendo un buen método.

Detección molecular de agentes infecciosos de transmisión sexual en un grupo de hombres sintomáticos y su relación con la conducta sexual / Molecular detection of sexually transmitted agents in a symptomatic group of men and its relationship with sexual behavior

Background: Sexually transmitted infections (STIs) affect sexual and reproductive health of millions of men. Pathogens such as human papillomavirus (HPV), herpes simplex virus type 1 and 2 (HSV-1 y HSV-2), Chlamydia trachomatis,Mycoplasmagenitalium,Mycoplasma hominis and Ureaplasma urealyticum are associated with STIs. Aim: To detect pathogens associated with STIs in symptomatic men and its relationship with sexual behavior. Methodology: DNA was obtained from exfoliated cells of penis from 20 symptomatic men. Pathogens were detected using qPCR or PCR followed by reverse line blot. Sexual behavior was evaluated through a survey. Results: Two or more infectious agents were detected in 50% of samples. U. urealyticum was found in 25%, meanwhile C. trachomatis and M. hominis were detected in 15%. VHS-1, VHS-2 andM. genitalium were detected only in 5%. HPV was found in all samples. The most frequent HPV genotypes were VPH 16, 11, 70. There were no statistical link found between sexual behavior and the studied microorganisms Conclusion: Infectious agents associated with STIs were detected in symptomatic men. HPV was the most frequent pathogen and it was detected in multiple genotypes. It is necessary to increase the sample size to associate significantly the sexual behavior with the results.

Introducción: Las infecciones de transmisión sexual (ITS) afectan la salud sexual y reproductiva de millones de hombres. Patógenos como virus papiloma humano (VPH), virus herpes simplex (VHS-1 y VHS-2), Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis y Ureaplasma urealyticum están asociados a ITS. Objetivo: Detectar patógenos asociados a ITS en hombres sintomáticos y relacionarlos con su conducta sexual. Metodología: Se obtuvo ADN de exfoliado celular del pene de 20 hombres sintomáticos de ITS. Los patógenos fueron detectados por RPC cuantitativa o RPC seguida de reverse line blot. La conducta sexual se evaluó mediante una encuesta. Resultados: En 50% de las muestras se detectaron dos o más agentes infecciosos; U. urealyticum fue detectado en 25% de los casos, mientras que C. trachomatis y M. hominis en 15%. VHS-1, VHS-2 y M. genitalium sólo en 5%. VPH se encontró en todas las muestras y los genotipos más frecuentes fueron VPH 16, 11, 70. No se encontró relación estadística entre los microorganismos estudiados y la conducta sexual de los encuestados. Conclusión: Se detectaron agentes infecciosos asociados a ITS en hombres sintomáticos, siendo VPH el más frecuente y encontrándose en múltiples genotipos. Es necesario aumentar el tamaño de muestra para asociar significativamente la conducta sexual a los resultados.