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.

Respuesta humoral y consecuencias reproductivas en ovejas desafiadas con Brucella ovis al final de la gestación / Immune response and reproductive consequences in experimentally infected ewes with Brucella ovis during late pregnancy

La brucelosis ovina por Brucella ovis es una enfermedad de prevalencia alta en Argentina. Para evaluar la patogenicidad de B. ovis y la respuesta serológica durante el último mes de gestación, 6 ovejas se distribuyeron en dos grupos: G1, ovejas preñadas, n = 4 y G2, ovejas no preñadas, n = 2. Tres ovejas del G1 (15 días preparto) y una del G2 fueron inoculadas con B. ovis. Se analizaron muestras de suero mediante diferentes pruebas serológicas. Se realizó aislamiento y PCR a partir de mucus cérvico-vaginal (mcv), placenta y leche. En las muestras de placenta se realizó histopatología. Las hembras del G1 parieron corderos vivos; se detectaron anticuerpos en las ovejas desafiadas del G1 a partir de los 5 días posinoculación. El mcv de las ovejas desafiadas resultó negativo al aislamiento en ambos grupos. Las muestras de leche del G1 fueron positivas por cultivo y PCR a B. ovis. La técnica de PCR resultó positiva en las placentas de las ovejas desafiadas del G1. La histopatología reveló una placentitis necrótica supurativa en una de las ovejas desafiadas. El desafío con B. ovis preparto resultó en la invasión de la placenta y de la glándula mamaria, con la consecuente excreción de la bacteria por leche. La infección con B. ovis indujo una respuesta humoral temprana en las ovejas. La colonización de la placenta por B. ovis y la excreción de la bacteria por la leche sugieren un potencial riesgo de infección activa para los corderos y la posibilidad de que estos se comporten como portadores latentes de la infección.

Ovine brucellosis by Brucella ovis is a highly prevalent disease in Argentina. This study aimed to evaluate the pathogenicity of B. ovis and the serological response in ewes during late pregnancy and in their offspring. Six adult ewes were distributed in two groupsGI (pregnant females, n = 4) and G2 (nonpregnant females, n = 2). Three pregnant ewes at 15 days prepartum and one nonpregnant eve were inoculated with B. ovis. Sera of sheep and their offspring were analyzed by different serological tests. Samples of cervicovaginal mucus, placenta and milk were studied by bacteriology. A Brucella genus-specific PCR assay was carried out in placenta and milk samples. Placenta samples were hystopathologically processed. G1 females gave birth to live lambs, but one died hours postpartum. Serological techniques employed detected antibodies in serum of inoculated pregnant animal 5 days postchallenge. Sera of female controls G1 and G2 remained negative throughout the study. Cervicovaginal mucus of infected ewes in G1 and G2 yielded negative results to bacteriology, but B. ovis was isolated from milk. The PCR assay was positive for the placenta and milk from inoculated pregnant ewes. Histopathology revealed necrotic suppurative placentitis in one placenta. However, although results demonstrated that B. ovis can invade the placenta and mammary gland, this bacterium did not cause abortion when it was inoculated intravenously at 15 days prepartum. B. ovis infection induced an early humoral response in pregnant ewes, but their lambs remained seronegative, indicating that there was no transfer of antibodies in infancy. Placenta colonization and milk excretion of B. ovis involves a potential source of infection for lambs, which could play a role as latent carriers of infection.

Accuracy of Nugent's score and each Amsel's criteria in the diagnosis of bacterial vaginosis.

OBJECTIVE: To determine the diagnostic accuracy of Nugent's score and each Amsel's criterion in the diagnosis of bacteria vaginosis (BV), considering Amsel's criteria as the gold standard. DESIGN: Cross-sectional, descriptive study (diagnostic test) Setting: Family planning clinic, Siriraj Hospital, Mahidol University. SUBJECTS: A total of 217 women who attended the Family Planning Clinic at Siriraj Hospital between August and December 2003. METHOD: Pelvic examination was performed on each participant. Samples of vaginal discharge was tested for BV infection using both Amsel's criteria and Nugent's score. Interpretation was made blinded without knowledge of each test result. Using Amsel's criteria as a gold standard, sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of Nugent's score and each of Amsel's criteria were estimated. RESULTS: Considering Amsel's criteria as the gold standard, Nugent's score showed a sensitivity of 65.6% (95%CI 46.8%, 80.8%), specificity of 97.3% (95%CI 93.5%, 99.0%), positive predictive value (PPV) of 80.8% (95%CI 60.0%, 92.7%), negative predictive value (NPV) of 94.2% (95%CI 89.7%, 96.9%) and accuracy of 92.6% (95%CI 88.1%, 95.6%). Both vaginal pH and whiff test demonstrated 100% sensitivity. However, vaginal pH showed lower specificity than the whiff test (58.9% and 97.3% respectively). CONCLUSION: Nugent's score might not be suitable to use as a screening test for diagnosis of BV due to its low sensitivity. The whiff test is the best clinical criteria of Amsel's criterion in the diagnosis of BV due to its high sensitivity and specitivity.

Asymptomatic shedding of genital herpes simplex virus [HSV-2] during pregnancy and neonatal outcome

The objectives of this study were detection of the asymptomatic shedding of genital herpes simplex virus type 2 [HSV-2] in the cervical secretion among the pregnant women late in pregnancy, through viral isolation by tissue culture [TC] and correlating it to the other methods of diagnosis or HSV-2, and also correlating it to the neonatal outcome. This study was conducted on 90 pregnant women, during the 3rd trimester who were coming for antenatal care, with no history, and no clinical findings of genital herpes. Asymptomatic HSV-2 shedding was determined by TC and polymerase chain reaction [PCR] of the cervical secretion, and the serologic status was determined by detecting HSV-2 IgM antibody. All of them were followed till delivery. An oropharyngeal swab of the neonates was taken for viral isolation by TC, and cord blood sample for HSV-IgM antibody. We found that: the percent of positivity of HSV-2 isolation by TC increased by increasing age, parity and number of previous abortions. [2] Asymptomatic shedding was detected by TC in 44/90 [48.9%] of the cervical swabs, HSV-2 DNA was detected in 43/44 [97.7%] of the +ve cervical swab, while serum HSV-2 IgM antibody was detected in 16/89 [18%] serum samples, and these represented 16/44 [36.4%] and 16/43 [37.2%] of the women with HSV-2 +ve TC and HSV-2 DNA detected by PCR respectively. [3] Only 3/90 [3.3%] oropharyngeal swabs showed HSV-2 +ve TC, their mothers were also HSV-2 +ve TC, while HSV-2 DNA, and HSV-2 IgM antibody could not be detected. [4] There was a significant correlation between HSV-2 isolation by TC and the perinatal outcome e.g. intrauterine fetal death [IUFD] still birth [SB], Apgar score, neonatal weight and number of neonates transferred to the neonatal intensive care unit [NICU], [5] There was a highly significant correlation between HSV-2 isolation by TC and HSV-2 DNA detection by PCR; while measurement of HSV-2 IgM antibody is of limited value in relation to both of them. HSV-2 isolation by TC is accurate, sensitive technique of low cost but of long turnaround time for diagnosis of asymptomatic shedding of HSV-2 during pregnancy. HSV-2 DNA detection by PCR can replace TC as it is more rapid but more expensive, and the cost benefit should be evaluated. So, TC is recommended to be used for screening HSV-2 infection, while PCR used for rapid diagnosis when subsequent intervention is essential e.g. antiviral treatment We could not conclusively relate these neonatal outcome to the asymptomatically shedded HSV-2

Aislamiento de la bacteria Ureaplasma sp. en el tracto reproductivo de vacas lecheras de Costa Rica / Isolation of the bacteria Ureaplasma sp. in the reproductive tract of milking cows in Costa Rica

This is the first report of Ureaplasma sp. from the reproductive tract of Costa Rican cows. Among 204 animals sampled from 11 dairy farms in the country's Central Plateau, the infection rate was 0-71 per cent. Isolation was more frequent in vulvo-vestibular (38.7 per cent) than in cervical swabs (23 per cent). Ureaplasma was correlated with clinical granular vulvitis symptoms

Chlamydia trachomatis: importância da colheita endocervical na diagnóstico pelo método de Papanicolaou / Chlamydia trachomatis: importance of the endocervical sampling in the diagnosis by the Papapicolaou method

Foram examinados citologicamente, pelo método de Papanicolaou e pela imunofluorescência direta (IF) para Chlamydia trachomatis (Ct), 161 esfregaços cervicovaginais de pacientes atendidas pelo Instituto Adolfo Lutz, Laboratório I de Taubaté (SP). Trinta e três casos (20,5%) tiveram teste positivo para Ct pel IF. Pelo método de Papanicolaou, foram considerados sugestivos de infecçäo por Ct os casos que apresentavam fina vacuolizaçäo nas células endocervicais metaplásicas, segundo critério recentes. O método de Papanicolaou, comparado a IF, mostrou alta sensibilidade (91,6%) no grupo de esfregaços que continham material endocervical. A IF é método de alta sensibilidade e especificidade, mas de custo näo acessível a populaçäo em geral. Dada a alta incidência de infecçöes por Ct no trato genital feminino, concluímos que o método de Papanicolaou em esfregaços contendo material endocervical continua sendo a primeira opçäo de exame para detecçäo deste agente

Mycoplasma hominis e ureaplasma urealyticum: pesquisa no material cervical de gestantes / Mycoplasma hominis and Ureaplasma urealyticum: research in pregnants cervical mucus specimens

Foram feitas pesquisas de Mycoplasma hominis e de Ureaplasma urealyticum em material cervical de gestantes. A identificaçäo de Ureaplasma urealyticum baseou-se nas provas bioquímicas das atividades da urease e da fosfatase e na observaçäo da morfologia colonial característica. Mycoplasma hominis foi caracterizado pela observaçäo da morfologia colonial de "ovo-frito", pelo comportamento bioquímico frente às seguintes provas: utilizaçäo da arginina, determinaçäo da atividade da fosfatase, reduçäo do tetrazólio em aerobiose e anaerobiose, utilizaçäo da glicose, produçäo de filmes e "spots", e foi identificado sorologicamente através das reaçöes de inibiçäo metabólica e de imunodifusäo dupla. Todos os micoplasmas isolados foram inicialmente submetidos à prova de sensibilidade à digitonina, 1,5% em soluçäo alcoólica, antes de serem submetido às provas de identificaçäo. Dois grupos de gestantes foram estudados: grupo de risco, constituído de 37 gestantes que possuíam histórico anterior de alteraçöes perigestacionais tais como: abortos, natimortos, prematuros, etc... e grupo controle constituído por 37 gestantes com gestaçöes anteriores normais. As taxas de colonizaçäo encontradas foram:73,0% de Ureaplasma urealyticum e 21,6% de Mycoplasma hominis no grupo de risco; no grupo controle os resultados foram: 65,0% de Ureaplasma urealyticum e 16,2% de Mycoplasma hominis. Estatisticamente as diferenças entre os dois grupos näo foram significantes (delta = 5%)

Modificaçäo e padronizaçäo de meios de transporte e cultivo de micoplasmas genitais: Mycoplasma hominis e Ureaplasma urealyticum / Modification and standardization of transport and culture media for genital mycoplasmas: Mycoplasma hominis and Ureaplasma urealyticum

Foram padronizados meios de transporte e de cultivo de micoplasmas genitais objetivando principalmente o cultivo do Mycoplasma hominis e do Ureaplasma urealyticum. Tais meios basearam-se principalmente nos preconizados por Shepard e também pelo Laboratory of Diagnosis of Mycoplasma Infections, (LDMI) aos quais foram introduzidas algumas modificaçöes com a finalidade de possibilitar a sua utilizaçäo na rotina laboratorial em nosso meio. A pesquisa desses microrganismos foi feita em material cervical de 74 gestantes. As amostras clínicas foram transportadas em meio líquido (MLTr) e cultivadas em meio líquido para ureaplasma (MLU) e meio líquido de arginina (MLA) para micoplasmas genitais que se utilizam da arginina como fonte energéntica, dentre eles o Mycoplamsa hominis. Os meios utilizados nas condiçöes aqui padronizados mostram-se bastante eficientes para a finalidade proposta, uma vez que nas gestantes as taxas de colonizaçäo encontradas para micoplasmas foram comparáveis com as citadas na literatura internacional

Vaginite por Gardnerella vaginalis: prevalência e avaliaçäo de metodologia para seu diagnóstico / Vaginitis by Gardnerella vaginalis: prevalence and evaluation of the methodology for its treatment

Gardnerella vaginalis foi isolada de 48,4% de 97 pacientes estudadas das quais 93,6% apresentavam um quadro da vaginite e/ou cervicite. Para seu isolamento foram utilizados os meios de Agar Sangue e Agar Vaginalis, ambos com sangue humano, incubados a 35§C por 72 horas, em lata com vela. A identificaçäo laboratorial foi baseada nos aspectos morfológicos macro e microscópicos, nas características tintoriais e nos testes de fermentaçäo de glicose, maltose e amido usando-se Cystine Trypticase Agar (CTA) como base. O material de 79 pacientes foi submetido a três técnicas diferentes de exame direto: a fresco, Gram e Papanicolau, encontrando-se uma correlaçäo com os achados de cultivo microbiológico em 36 (81,8%) das 44 culturas positivas. Células do fenômeno Gardner-Dukes, indicativos da presença de G. vaginalis foram encontradas em 32 (72,7%) dos exames positivos

Valor diagnóstico de hallazgos citomorfológicos de Clamydia tracomatis en extendidos cervicales

Cuatro mil seiscientos ochenta extendidos cervicales de rutina fueron estudiados prospectivamente durante un año y medio. Ciento veinte (2,6%) tuvieron cambios citológicos compatibles con infección por Clamydia tracomatis. Estos cambios fueron correlacionados con la edad, síntomas génito-urinarios, métodos anticonceptivos y enfermedades venéreas. Los extendidos fueron teñidos con el método de Papanicolau y luego teñidos con PAS. Todas las fases del ciclo de Clamydia tracomatis fueron teñidas. Del total de casos estudiados, 115 (95,8%) tenían células de metaplasia, 10 (8,3%) estaban asociadas con CIN y 10 (8,3%) con atipia coilocitótica. De estos últimos, 2 estaban asociados con CIN I, 2 con CIN II y 6 tenían atipia coilocitótica solamente

Prueba poscoital: correlación de la misma con los resultados del cultivo del moco cervical, del espermocultivo, del cultivo del líquido prostático y del espermograma / Postcoital test: its correlation with results from the culture of cervical mucus, spermoculture, culture of prostatic fluid and spermogram

Se hace un recuento histórico donde se pone de manifiesto el origen de la prueba poscoital o de Sims-Hünner, así como la importancia que el mismo ha tenido y aún tiene en el estudio de la pareja infértil. Se realiza un estudio de los resultados de los test poscoitales realizados a 50 parejas. Se correlacionan dichos resultados con los obtenidos del cultivo del moco cervical, del espermocultivo, del cultivo del líquido prostático y del espermograma

Transporte de la Escherichia coli por el espermatozoide humano a través del moco cervical mesocíclico / Escherichia coli transport by the human spermatozoa through the mesocyclic cervical mucous

Se sugiere que los Espermios humanos podrían actuar como vehículos de transporte de microorganismos: en nuestro trabajo hemos observado la unión específica de E. Coli a los mismos. La penetración de los espermios depende del estado del moco cervical durante el ciclo menstrual: Es máxima en el moco de tipo E (Estrogénico) y ausente en el moco de tipo G (Progestativo). Estudiamos la migración de los espermios a través del moco cervical Mesocíclico "In Vitro". La Microfotografía electrónica de barrido nos permitió observar una estrecha relación entre bacterias y espermios humanos. Nuestros resultados permiten apoyar la hipótesis de que este mecanismo mediado por espermios podría explicar el origen de procesos inflamatorios pelvianos