Academic literature on the topic 'Ureaplasmas'

Abstract Genital tract ureaplasma infections are associated with numerous complications, ranging from inflammation, through infertility, to problematic pregnancy. In the course of ureaplasma infection, the risk of human papillomavirus infection increases. Diagnostic tests for urea-plasma infections are not always carried out, especially in women with the normal Nugent test results. The study attempts to check whether it is possible to find a prognostic indicator that could suggest a high abundance of ureaplasmas (≥ 104 CFU/ml) at the stage of the initial examination of vaginal discharge. Such a prognostic factor could qualify women for further tests to detect infections with these atypical bacteria. Six hundred twenty-seven white women with a score of 0–3 on the Nugent scale were tested, including 322 patients with a high abundance of ureaplasmas (≥ 104 CFU/ml) and 305 who tested negative for these bacteria. Ureaplasma infections were detected statistically significant in women who had few or no epithelial cells in the genital swab specimens compared to the results obtained for women with numerous or very numerous epithelial cells (p < 0.001). The risk of the high density of ureaplasmas was 38.7% higher with fewer or no epithelial cells than with high numbers. In patients aged 18–40 years with few or no epithelial cells, a high density of ureaplasmas (≥ 104 CFU/ml) was observed significantly more frequently (p = 0.003). Determining the number of epithelial cells in Gram-stained slides may be the prognostic indicator of ureaplasma infection. Testing for genital ureaplasma infection should be considered, especially in women of childbearing age (18–40 years), even if the Nugent test value is normal and pH ≤ 4.6.

Bronchopulmonary dysplasia is associated with chorioamnionitis and fetal lung inflammation. Ureaplasma species are the bacteria most frequently isolated from chorioamnionitis. Very chronic ureaplasma colonization of amniotic fluid causes low-grade lung inflammation and functional lung maturation in fetal sheep. Less is known about shorter exposures of the fetal lung. Therefore, we hypothesized that ureaplasmas would cause an acute inflammatory response that would alter lung development. Singleton ovine fetuses received intra-amniotic Ureaplasma parvum serovar 3 or control media at 110, 117, or 121 days and were delivered at 124 days gestational age (term = 150 days). Inflammation was assessed by 1) cell counts in bronchoalveolar lavage fluid (BALF), and 2) cytokine mRNA measurements, immunohistochemistry, and flow cytometry for inflammatory cells and elastin and α-smooth muscle actin (α-SMA) staining in lung tissue. Neutrophils were increased in BALF 3 days after exposure to ureaplasmas ( P = 0.01). Myeloperoxidase-positive cells increased after 3 days ( P = 0.03), and major histocompatibility complex (MHC) class II-positive cells increased after 14 days of ureaplasma exposure ( P = 0.001). PU.1 (macrophage marker)- or CD3 (T lymphocyte marker)-positive cells were not induced by ureaplasmas. CD3-positive cells in the posterior mediastinal lymph node increased in ureaplasma-exposed animals at 3, 7, and 14 days ( P = 0.002). Focal elastin depositions decreased in alveolar septa at 14 days ( P = 0.002), whereas α-SMA increased in arteries and bronchioli. U. parvum induced a mild acute inflammatory response and changed elastin and α-SMA deposition in the lung, which may affect lung structure and subsequent development.

The clinical significance of Ureaplasmas in urogenital pathology is reviewed. Ureaplasmas belong to the class Mollicutes. Asymptomatic carriage of these bacteria is common, and most individuals do not develop disease. Ureaplasma urealyticum and Ureaplasma parvum are sexually transmitted bacteria among humans implicated in a variety of disease states including but not limited to: nongonococcal urethritis, adverse pregnancy outcomes, chorioamnionitis, and bronchopulmonary dysplasia in neonates. U. urealyticum has been associated with urethritis in men and is revealed in a high concentration that confirms its etiological role in the disease. Men with a high U. urealyticum load are considered for treatment, however, the data on the therapy efficiency have been insufficient so far. In symptomatic women, bacterial vaginosis should always be tested for, and the corresponding therapy should be prescribed in case of positive results.

SUMMARYAs an estimate of their virulence, the ability of ovine, bovine, canine, feline and simian ureaplasma strains to cause mastitis in the ovine mammary gland was investigated. Five ovine ureaplasmas produced a clinical mastitis. Broth cultures of seven bovine ureaplasmas were unable to infect the ovine gland, but two of these strains plus one other were able to do so following passage through the bovine udder. One of two canine strains and a feline strain both caused mastitis, but the simian strain persisted at low titre for only 5 days post-inoculation in one of the two ewes tested.

SUMMARY The genital mycoplasmas represent a complex and unique group of microorganisms that have been associated with a wide array of infectious diseases in adults and infants. The lack of conclusive knowledge regarding the pathogenic potential of Mycoplasma and Ureaplasma spp. in many conditions is due to a general unfamiliarity of physicians and microbiology laboratories with their fastidious growth requirements, leading to difficulty in their detection; their high prevalence in healthy persons; the poor design of research studies attempting to base association with disease on the mere presence of the organisms in the lower urogenital tract; the failure to consider multifactorial aspects of diseases; and considering these genital mycoplasmas only as a last resort. The situation is now changing because of a greater appreciation of the genital mycoplasmas as perinatal pathogens and improvements in laboratory detection, particularly with regard to the development of powerful molecular nucleic acid amplification tests. This review summarizes the epidemiology of genital mycoplasmas as causes of neonatal infections and premature birth; evidence linking ureaplasmas with bronchopulmonary dysplasia; recent changes in the taxonomy of the genus Ureaplasma; the neonatal host response to mycoplasma and ureaplasma infections; advances in laboratory detection, including molecular methods; and therapeutic considerations for treatment of systemic diseases.

ABSTRACT The in vitro susceptibilities of 151 unique clinical isolates of Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma genitalium, and Ureaplasma species to DC-159a, an investigational fluoroquinolone, in comparison with those to other agents were determined. Macrolides were the most active agents against M. pneumoniae and M. genitalium, whereas clindamycin was most active against M. hominis. DC-159a MICs were ≤0.5 μg/ml for all Mycoplasma species and ≤4 μg/ml for ureaplasmas. DC-159a was the most active fluoroquinolone tested against M. pneumoniae and M. fermentans, and it was second to moxifloxacin against the other species. It was bactericidal against 10 M. pneumoniae isolates and demonstrated killing of ≥99.9% of the inoculum at 24 h for 2 isolates. The excellent in vitro activity of DC-159a demonstrates its potential for use in the treatment of infections due to mycoplasmas and ureaplasmas.

We investigated the influence of symptoms and signs on the detection of Chlamydia trachomatis, Mycoplasma genitalium and Ureaplasma urealyticum organisms (ureaplasmas) in men with non-gonococcal urethritis (NGU). Two hundred and forty-two men attending the Jefferiss Wing at St Mary's Hospital for a sexual health assessment were evaluated, of whom 169 had NGU. Urethral inflammation was diagnosed if there were either ≥5 polymorphonuclear leucocytes (PMNLs) per high-power field (HPF) in five or more microscope fields of a Gram-stained urethral smear, or ≥10 PMNLs per HPF in five or more fields of a Gram-stained thread from 15-20 mL of a first-passed urine (FPU) specimen. C. trachomatis was diagnosed by direct immunofluoresence, M. genitalium by a polymerase chain reaction assay and ureaplasmas by culture. On multivariate analysis, to control for potential confounding by age, ethnicity, sexual lifestyle and co-infection, an urethral discharge remained significantly associated with the detection of C. trachomatis and M. genitalium in men with acute urethritis [OR 12.3, 95% CI (2.39-63.5) and OR 35.2, 95% CI (3.9-319.6), respectively], but dysuria or penile irritation did not. The detection of ureaplasmas was not associated with any clinical feature. In addition, on multivariate analysis men with NGU who were either symptomatic or had an observable discharge were more likely to have C. trachomatis or M. genitalium detected [(OR 6.92, 95% CI 1.41-33.9) and (OR 5.18, 95% CI 0.99-27.1), respectively], but not ureaplasmas (OR 1.19, 95% CI 0.33-4.35). The findings suggest that in men with acute NGU, symptoms or signs, and in particular a urethral discharge, are associated with the detection of C. trachomatis and M. genitalium, but not ureaplasmas. Currently, there is no precise answer to the question of whether all men attending a GUM clinic need to be screened for NGU, but if clinically asymptomatic NGU is found not to be associated with a sexually transmitted pathogen, the UK clinical guidelines requiring the preparation of a urethral smear from such men would need to be revised.

ABSTRACT The susceptibilities of Mycoplasma hominis,Mycoplasma pneumoniae, and Ureaplasma urealyticum to eight new antimicrobial agents were determined by agar dilution.M. pneumoniae was susceptible to the new glycylcycline GAR-936 at 0.12 μg/ml and evernimicin at 4 μg/ml, but it was resistant to linezolid. It was most susceptible to dirithromycin, quinupristin-dalfopristin, telithromycin, reference macrolides, and josamycin. M. hominis was susceptible to linezolid, evernimicin, and GAR-936. It was resistant to macrolides and the ketolide telithromycin but susceptible to quinupristin-dalfopristin and josamycin. U. urealyticum was susceptible to evernimicin (8 to 16 μg/ml) and resistant to linezolid. It was less susceptible to GAR-936 (4.0 μg/ml) than to tetracycline (0.5 μg/ml). Telithromycin and quinupristin-dalfopristin were the most active agents against ureaplasmas (0.06 μg/ml). The new quinolone gatifloxacin was active against M. pneumoniae and M. hominis at 0.12 to 0.25 μg/ml and active against ureaplasmas at 1.0 μg/ml. The MICs of macrolides were markedly affected by pH, with an 8- to 32-fold increase in the susceptibility of M. pneumoniae as the pH increased from 6.9 to 7.8. A similar increase in susceptibility with increasing pH was also observed with ureaplasmas. Tetracyclines showed a fourfold increase of activity as the pH decreased 1 U, whereas GAR-936 showed a fourfold decrease in activity with a decrease in pH.

Foram utilizadas 112 amostras de muco vulvovaginal, coletados de vacas com distúrbios reprodutivos, para pesquisa de Mycoplasma e Ureaplasma. Para isolamentos, foram usados meios específicos para micoplasmas (SP-4) e para ureaplasmas. PCR genérica, PCR específica para Mycoplasma bovis e nested-PCR em tubo único para Ureaplasma diversum foram realizados com os DNAs extraídos das amostras. Mycoplasma spp. e U. diversum foram detectados em 12,5 e 25,0%, respectivamente. A PCR genérica resultou em reações positivas em 63,4% das amostras transportadas em SP-4 e em 69,6% das transportadas em meio de ureaplasma. M. bovis foi detectado, na PCR específica, em 9,8% das amostras e U. diversum, na nested-PCR, em 37,5%. Houve maior sensibilidade na metodologia da PCR quando comparada à técnica de cultivo para Mycoplasma e Ureaplasma.

Dissertação para obtenção do grau de Mestre em Biologia Molecular em Saúde
Neste estudo foram analisados os dados referentes às bactérias Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum e Ureaplasma parvum, duma amostra constituída por 16 020 utentes de um laboratório de análises clínicas. Constatou-se que o maior número de pedidos de análises recaía sobre a Chlamydia trachomatis, dado esta ser uma das doenças sexualmente transmissíveis com maior prevalência atualmente, e que a faixa etária com maior pedido de análises se situa entre os 27 e os 34 anos, correspondendo a adultos sexualmente ativos e em idade reprodutiva. De salientar também a elevada incidência de infertilidade em Portugal e a ligação entre este facto e as doenças sexualmente transmissíveis. Verificou-se que do total das análises requeridas apenas 4,6% foram amostras do género masculino. No entanto, são os homens que apresentam globalmente uma maior taxa de positivos, estando este facto associado à existência de sintomatologia aquando do diagnóstico. Da análise da amostra em estudo conclui-se que as colheitas de urina e exsudado uretral são os produtos que apresentam maior positividade para ambos os sexos, no entanto, a colheita com maior número de pedidos é a de exsudado vaginal. Foi possível ainda verificar que os métodos moleculares que permitem a distinção entre ureaplasmas contribuem para o uso racional de antibióticos.

Group B streptococcus (GBS, S. agalactiae) and human ureaplasmas (U. parvumand U. urealyticum) are two clinically and phylogenetically related, potential perinatal pathogens. Their relationships between genotypes and pathogenesis of GBS and ureaplasma infection were still not well understood, one of the reason is that both of them are still short of a very practical genotyping system. In the study, to solve the above problem we developed genotyping systems for the organisms (the second section). For human ureaplasmas, based on four genes/gene clusters (rRNAgene clusters, the elongation factor Tu genes, urease gene complexes and multiplebanded antigen genes), we designed many primer pairs suitable for developing species identification assays for the two newly established human ureaplasma species (U. parvum and U. urealyticum). Further, based on the heterogeneity of ureaplasma multiple banded antigen gene (which contains species- and serovar-specific regions), we developed genotyping methods for each ureaplasma species.For GBS, based on three sets of molecular markers (capsular polysaccharidesynthesis gene clusters, surface protein antigen genes and mobile genetic elements),we developed a genotyping system. The primary evaluation of the genotyping systems showed that the genotyping systems were practical alternative assays for the conventional serotyping and they will be useful to further explore the relationships between genotypes and pathogenesis of GBS and ureaplasma infection. In the study, we introduced novel data and tools into GBS and ureaplasma studies especially from genomic- and bioinformatics-based molecular microbiology(the third section). For two newly established human ureaplasma species, based on the U. parvum serovar-3 genome, and using the above four important genes/geneclusters, we exposed some interesting problems in the understanding of newureaplasma taxonomy especially in the post genomic era. For GBS, we studied the two published full genomes and exposed some new problems or possible future new research fields. In particular we found the two finished and one ongoing GBS genomes were all non-typical and suggest that future genomic project had better have genetic population structure viewpoint. Finally, we suggested that integrated studies of the two potential or conditional perinatal pathogens, from the viewpoint of evolution, would provide a new understanding angle of the pathogenesis of the two organisms. Studies suggested that during coevolution, human ureaplasmas(especially U. parvum) became friendlier than their ancestors to their human host (by losing most of its virulence genes); however, GBS tried to increase its invasive abilities (by getting more virulence genes) to fight against the human host attack.

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CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico
Mycoplasmas are microorganisms lacking cell walls with reduced ability Biosintética, which makes them extremely fastidious growth. The strength of these microorganisms to antimicrobials used in his therapy as tetracyclines, macrolides and quinolones have been reported with increased frequency. The determination of antimicrobial susceptibility is particularly difficult because not shown in broth turbidity which complicates the standardization of the inoculum and are extremely susceptible pH conditions. The method most commonly used for this purpose is based on metabolic inhibition using specific substrates. This study evaluated the susceptibility of isolates of Mycoplasma hominis and Ureaplasma sp. stored in the period from 2011 to 2012. The methodology used for the isolation comprised culture techniques, for storage and freezing of the strains we used selective broth enriched PPLO and BHI, Thawing of isolates was succeeded subculture in broth enriched selective and differential agar A7 for phenotypic characterization. Quantification, identification and antimicrobial susceptibility testing was performed using the the triage system Mycofast Evolution ® Screening (Elitech Group), whose identification is based on bacterial susceptibility by Identibiotiqué system. The susceptibility of the isolates was determined against antimicrobial agents Doxiciclina 8 μg/ml, Roxitromicina 4 μg/ml and Ofloxacina 4 μg/ml, widely used in the empirical treatment of patients with urogenital tract infections. Our study showed high level of resistance of the isolates to doxicycline and ofloxacin. This is the first study involving isolation and susceptibility profile of these agents undertaken in the Amazonia.
Micoplasmas são microrganismos desprovidos de parede celular, com reduzida capacidade Biossintética, o que os torna extremamente fastidiosos ao crescimento. A resistência destes microrganismos aos agentes antimicrobianos utilizados na sua terapia como as tetraciclinas, macrolídeos e quinolonas tem sido relatadas com frequência cada vez maior. A determinação da susceptibilidade a antimicrobianos é particularmente difícil porque não mostram turbidez em caldo o que dificulta a padronização do inóculo e são extremamente susceptíveis as condições de pH. A metodologia utilizada para este fim baseia-se na inibição metabólica utilizando substratos específicos. O presente trabalho avaliou a susceptibilidade de isolados de Mycoplasma hominis e Ureaplasma sp. arrmazenados no período de 2011 a 2012. A metodologia utilizada englobou técnicas de cultura; para a estocagem e congelamento das cepas empregou-se caldo seletivo enriquecido PPLO e BHI, O descongelamento dos isolados foi sucedido de subcultivos em caldos enriquecidos seletivos diferenciais e Agar A7 para caracterização fenotípica. A quantificação, a identificação e o teste de susceptibilidade a antimicrobianos foi realizada através do sistema de triagem Mycofast® Screening Evolution (Elitech Group), cuja identificação se baseia na susceptibilidade bacteriana pelo sistema Identibiotiqué. A susceptibilidade dos isolados foi determinada frente aos antimicrobianos doxiciclina 8 μg/ml, roxitromicina 4 μg/ml e ofloxacina 4 μg/ml, amplamente utilizados no tratamento empírico de pacientes com infecções do trato urogenital. Nosso estudo detectou alto nível de resistência dos isolados armazenados para doxiciclina e ofloxacina. Este é o primeiro estudo envolvendo isolamento e o perfil de susceptibilidade destes agentes realizado na região norte.

Bibliography
Thesis (PhD (Pathology. Medical Microbiology))--University of Stellenbosch, 2010.
ENGLISH ABSTRACT: Overview: Mycoplasmas and ureaplasmas are not routinely diagnosed and are under researched in South Africa. Prevalence, population shifts especially concerning genital flora and implications in infection or other conditions are unknown. Information pertaining to Mycoplasma pneumoniae in respiratory disease is similarly lacking. There is little information on antimicrobial susceptibilities and resistance development against Sexually Transmitted Infections (STI) syndromic management approaches. Aims: a) Elucidate mycoplasmal and ureaplasmal prevalence and contributing factors concerning cervical colonisation or preterm delivery in conjunction with HIV and Chlamydia trachomatis b) Investigate prevalence of M. pneumoniae in respiratory infections in conjunction with HIV, Mycobacterium tuberculosis and Pneumocystis jiroveci. c) Determine antimicrobial susceptibilities of mycoplasmas and ureaplasmas and analyse resistance genes. d) Assess the inter-generic transfer potential of resistance gene (tetM) between Ureaplasma spp. and Neisseria gonorrhea. Genital setting: The prevalence of genital mycoplasmas, ureaplasmas and Chlamydia on women attending their first prenatal visit, in conjunction with preterm labour or HIV status was investigated. For preterm labour (2003), 199 women were monitored for preterm delivery (<37 weeks); for colonisation and HIV (2005), 219 women were screened. Microbial detection was performed on DNA extracted from endocervical swabs employing PCR techniques. Colonisation was seen to be highest in the 14-20 year group from 2003. In women aged ±21 years, co-colonisation was 13% although there was a shift from co-colonisation with Mycoplasma hominis and Ureaplasma spp. in 2003 to other dual/triple combinations in 2005. Overall major trends from both collection periods were that the prevalence of Ureaplasma spp. tended to be higher in women ±26 years, whilst prevalence of C. trachomatis and M. hominis were lower. No association was evident between colonisation with M. hominis, U. urealyticum, Ureaplasma parvum and labour outcome. HIV status had no effect on the prevalence/co-colonisation of M. hominis, Ureaplasma spp. or C. trachomatis. Respiratory setting: Studies were conducted to determine the prevalence of community acquired atypical pneumonias in adults (M. pneumoniae and P. jiroveci) and neonates (mycoplasmas, ureaplasmas and Chlamydia trachomatis) in order to improve treatment management programmes in the Port Elizabeth region. Sputum specimens from 102 adult patients presenting with pneumonia/symptoms of pneumonia admitted to hospitals were assessed by PCR. Details of patient’s gender, age, HIV and Mycobacterium tuberculosis status were provided by the hospitals. Women were seen to be at high risk for community-acquired P. jiroveci colonisation. Overall, prevalence of P. jiroveci was 52.9% (54/102 patients). P. jiroveci was mainly associated with HIV (25/74) (P. jiroveci and HIV positive patients in patient sample for which clinical data and HIV status was available) and co-infection with M. tuberculosis was observed in 12 HIV cases and one HIV negative patient. No DHPS (20) or DHFR (17) resistance associated mutations were found in P. jiroveci. M. pneumoniae was detected in one patient. For prevalence studies (2007-2008) on atypical pneumonia in neonates, 69 endotracheal aspirates were obtained. PCR detection of M. hominis, U. urealyticum and C. trachomatis was performed and U. parvum detected in two specimens. Antibiotic susceptibilities and resistance genes: The following investigations on clinical isolates of U. parvum and U. urealyticum were conducted (i) antibiotic susceptibility profiles, (ii) detection of drug target gene mutations, or gene acquisitions and (iii) inter-generic resistance gene transfer potential to Neisseria gonorrhoeae. Culture techniques applied to 132 endocervical specimens provided 66 Ureaplasma cultures (35 U. parvum, 9 U. urealyticum, 22 U. parvum + U. urealyticum). MIC determinations to ofloxacin, erythromycin, tetracycline, doxycycline, azithromycin and josamycin were performed. Thirty-seven ureaplasma cultures were fully susceptible to all antibiotics tested; 21 showed intermediate resistance to erythromycin, azithromycin and ofloxacin; while seven were resistant to tetracycline, three of which were also resistant to doxycycline and one also resistant to azithromycin. Concerning ofloxacin resistance directed at quinolone resistance determining regions, a substitution of Ser83Leu in ParC was demonstrated in one intermediately-resistant Ureaplasma (MIC 4 µg/ml) while a triple substitution of Asp112Glu in GyrA along with Ala125Thr and Ala136Thr in ParC was found in six further intermediately-resistant strains. No mutations were found in strains with MICs 1 µg/ml. No mutations were detected in 23S rRNA operons, L4 or L22 proteins. TetM and int-Tn genes were found in seven tetracycline-resistant strains. On screening 59 tetracycline-susceptible and -intermediate strains, eleven whilst possessing an int-Tn gene lacked a large region of tetM and 48 only contained small regions of tetM. The tetM genes of the seven tetracycline-resistant strains were sequenced and comparisons performed against GenBank sequences of Neisseria gonorrhoeae, Streptococcus pneumoniae and U. urealyticum. For five strains tetM was seen to be highly mosaic in structure containing regions that were similar to those of the GenBank strains and others that were unique. In the tetM leader region, four hot spot recombination sites were identified that could certainly influence the formation of the mosaic structures, upstream insertion sequences/open reading frames and transposon regions that regulate expression. On characterising the int-Tn genes of the seven tetracycline-resistant strains, three types were present indicating transposons from different origins had integrated into ureaplasma genomes. Reciprocal tetracycline resistance gene transfer between ureaplasmas and N. gonorrhoeae were unsuccessful. However, low-level tetracycline resistance (MICs 4-8 µg/ml) was transferred to a U. parvum recipient from one U. urealyticum and three U. parvum donors that carried tetM with MICs 16-64 µg/ml. On tetM PCR analysis, tetM was not detected in the transformants. Conclusions: The importance of genital mycoplasmas, ureaplasmas and C. trachomatis in long term aetiologies requires further investigations, certainly in relation with syndromic management regimens that fail to reduce colonisation rates. The high prevalence of P. jiroveci, the presence of M. pneumoniae in cases of pneumonia and detection of U. parvum in two cases of neonatal pneumonia investigated emphasises that in the absence of definitive diagnoses, it is crucial to monitor treatment responses carefully, especially when first line antibiotic preferences are ß-lactams, in order to ensure adequate and informed delivery of medical care. The finding of transposon and/or tetM regions in all ureaplasmas investigated with or without full expression of tetracycline resistance, in conjunction with tetM gene diversity, certainly places ureaplasmas strongly in the picture for intra- and inter-generic exchange of antibiotic resistance genes.
AFRIKAANSE OPSOMMING: Oorsig: Mikoplasma en ureaplasma word nie roetineweg gediagnoseer nie en in Suid Afrika is nog min navorsing daaroor gedoen. Prevalensie, populasie verskuiwings, veral in genital flora, en die impliksies van infeksie en ander toestande is onbekend. Inligting rakende Mycoplasma pneumoniae in respiratoriese siekte is ook gebrekkig. Daar is min inligting beskikbaar rakende die antimikrobiale vatbaarheid en die ontwikkeling van weerstandigheid gesien teen die benadering tot sindromiese hantering van seksueel oordraagbare siektes. Doelwitte: a) Om inligting te verskaf oor die prevalensie van mikoplasma en ureaplasma en bydraende faktore betreffende voortydige kraam tesame met MIV en Chlamydia trachomatis. b) Ondersoek van die prevalensie van M. pneumoniae in respiratoriese infeksies tesame met MIV, Mycobacterium tuberculosis en Pneumocystis jiroveci. c) Bepaling van die antimikrobiale vatbaarheid van mikoplasma en ureaplasma en analisevan weerstandigheids gene. d) Bereken die inter-genetiese oordrag potensiaal van weerstandigheids gene (tetM) tussen Ureaplasma spp. en Naisseria gonorrhoeae. Genitale omgewing: Die prevalensie van genitale mikoplasma, ureaplasma en Chlamydia in vroue tydens hul eerste prenatale besoek, tesame met vroegtydige kraam en MIV status is ondersoek. In voortydige kraam (2003), is 199 vroue gemonitor vir voortydige kraam (<37 weke); vir kolonisasie en MIV (2005), is 219 vroue getoets. Mikrobiale toetsing is gedoen deur DNS te win vanaf endoservikale deppers met PKR tegnieke. Kolonisasie was die hoogste in die ouderdomsgroep 14.20 jaar, in 2003. In vroue van ±21 jaar was medekolonisasie 13% alhoewel daar en verskuiwing was van mede-kolonisasie met Mycoplasma hominis en Ureaplasma spp. in 2003 tot ander dubbel/trippel kombinasies in 2005. Die oorkoepelende tendens in altwee die tydperke van waarneming was dat die prevalensie van Ureoplasma spp. geneig was om hoër te wees in vroue ±26 jaar, terwyl prevalensie van C. trachomatis en M. hominis laer was. Geen assosiasie kon getoon word tussen koloniesasie met M. hominis, U. urealyticum, Ureaplasma parvum en uitkoms van kraam nie. MIV status het geen effek gehad op die prevalensie/mede-kolonisasie van M. hominis, Ureaplasma spp. of C. Trachomatis nie. Respiratories: Studies is gedoen om die prevalensie van gemeenskaps verworwe atipiese pneumonie in volwassenes (M. pneumoniae en P. jiroveci) en neonate (mikoplasma, ureaplasma en Chlamydia trachomatis) te bepaal om behandeling en hantering programme in die Port Elizabeth area te verbeter. Sputum monsters van 102 volwasse pasiënte wat presenteer het met pneumonie of simptome van pneumonie en wat tot hospitale toegelaat was, is ontleed. Besonderhede van die pasiënte se geslag, ouderdom, MIV en Mycobacterium tuberculosis status is deur die hospitale verskaf. PKR is gedoen met inleiers gerig teen die volgende gene: P. jiroveci vir die aantoning van mitokondriale groot subeenheid RNS en vir die analise van mutasies vir ko-trimoksasool weerstandigheid dihydropteroaat sintetase (DHPS) en dihydrofolaat reduktase (DHFR); M. pneumoniae vir die aantoning van P1 adhesien en 16S rRNS. Vroue het ‘n hoë risiko vir gemeenskapsverworwe P. jiroveci kolonisasie gehad. In die algemeen was die prevalensie van P. jiroveci 52.9% (54/102 pasiënte). P. jiroveci was hoofsaaklik geassosieerd met MIV (25/74) (P. jiroveci en MIV positiewe pasiënte in die pasiënt monster waarvoor daar kliniese data en MIV status bekend was) en mede-infeksie met M. tuberculosis is gesien in 12 MIV gevalle en een MIV negatiewe pasiënt. Geen DHPS (20) of DHFR (17) weerstandigheids geassosieerde mutasies is gevind in P. Jiroveci nie. M. pneumoniae was aangetoon in een pasiënt. Vir prevalensie studies (2007-2008) op atipiese pneumonie in neonate is 69 endotrageale aspirate verkry. PKR toetsing vir M. hominis, U. urealyticum en C. trachomatis is gedoen met ‘primers’ soos voorheen gepubliseer. Ureaplasma parvum is aangetoon in twee neonate met PKR met negatiewe kultuur resultate. Antibiotika sensitiwiteite en weerstandigheids gene: Die volgende toetse is gedoen op kliniese isolate van U. parvum en U. urealyticum (i) antibiotika sensitiwiteits profiele, (ii) aantoning van teiken geen mutasies, of geen aanwinste en (iii) potensiaal vir inter-generiese weerstandigheids geen oordrag na Neisseria gonorrhoeae. Kultuur tegnieke toegepas op 132 endoservikale monsters het 66 Ureaplasma kulture gelewer (35 U. parvum, 9 U. urealyticum, 22 U. parvum + U. urealyticum). MIK bepaling vir ofloksasien, eritromisien, tetrasiklien, doksisiklien, azitromisien en josamisien is gedoen. Sewe-en-dertig kulture was ten volle sensitief vir alle antibiotika wat getoets is; een-en twintig het intermediere weerstandigheid teenoor eritromisien, azitromisien en ofloksasien getoon, terwyl sewe weerstandig was vir tetrasiklien, drie daarvan was ook weerstandig vir doksisiklien. Wat betref ofloksasien weerstandigheid gemik teen kwinoloon weerstandigheids bepalende gebiede, is vervanging van Ser83Leu in ParC gedemonstreer in een intermedier weerstandige Ureaplasma (MIK 4 µml) terwyl en trippel vervanging van Asp112Glu in GyrA saam met Ala125Thr en Ala136Thr in ParC gevind is in ses ander intermedier weerstandige stamme. Geen mutasies is gevind in stamme met MIKs van MICs 1 µg/ml nie. Geeneen van die ureaplasma was weerstandig vir eritromisien/azitromisien nie en geen mutasies is gevind in 23S rRNA operons , L4 of L22 proteine nie. TetM en int- Tn gene is gevind in sewe tetrasiklien weerstandige stamme. 58 Tetrasiklien sensitiewe en .intermediere stamme is getoets, waarvan elf en int-Tn geen gekort het sowel as en groot deel van tetM, terwyl 48 slegs klein dele van TetM bevat het. Die tetM gene van die sewe tetrasiklein-werstandige stamme se geenvolgorde is bepaal en vergelykings is getref teenoor die GenBank volgordes van Neisseria gonorrhoeae, Streptococcus pneumoniae en U. urealyticum. In vyf stamme is gevind dat die tetM geen hoogs mosaiek in struktuur was met areas wat ooreenstem met die in GenBank stamme, en ander areas wat uniek is. In die tetM leier area, is vier ehot spot f herkombinasie areas geidentifiseer wat sekerlik die vorming van die mosaiiek strukture kon beinvloed, asook transposon areas wat geenuitdrukking bepaal. Met karakterisering van die int-Tn gene van die sewe tetrasikleinweerstandlige stamme, was drie tipes teenwoordig waarin transposons vanaf verskillende oorsprong aangedui was, geintegreerd met die ureaplama genome. Resiprokale tetrasiklien weerstandigheids geen oordrag tussen ureaplasma en n. gonorrhoea was nie suksesvol nie. Lae-vlak tetrasiklien weerstandigheid (MIK fs van 4 . 8 µg/ml) is wel suksesvol oorgedra na en U. parvum ontvanger vanaf een U. urealyticum en drie U. parvum ontvangers wat tetM gedra het met MIKs van 16-64 µg/ml. Met die analise van tetM met PKR, kon tetM nie aangetoon word in die transformante nie. Gevolgtrekkings: Die belang van genitale mykoplasma, ureaplasma en C. trachomatis in langtermyn etologie benodig verdere ondersoek, veral in die lig van die sindromiese behandeling regimes wat nie kolonisasie verminder nie. Die hoe prevalensie van P. jiroveci, die teenwoordigheid van M. pneumoniae in gevalle van pneumonie en die aantoning van U. parvum in twee gevalle van neonatale pneumonie benadruk dat, in die afwesigheid van en definitiewe diagnose, dit noodsaaklik is om respons tot behandeling sorgvuldig te moniteer, veral indien die eerste lyn antibiotika keuse ß-laktam antimikrobiale middels of kefalosporiene is, sodat behoorlike en ingeligde gesondheidsorg gelewer kan word. Die bevinding van transposon en/of tetM gebiede in alle ureaplasma wat ondersoek is met of sonder volle uitdrukking van tetrasiklien weerstandigheid, in samehang met tetM diversiteit, plaas verseker ureaplasma sterk in die prentjie vir intra- en inter-generiese uitruiling van antibiotika weerstandigheids gene.
Nelson Mandela Metropolitan University
National Research Foundation (NRF Thuthuka)
Medical Research Council

Os Mollicutes causam doenças em várias espécies animais de importância econômica, inclusive em bovinos. Neste estudo, foi avaliada por concentração inibitória mínima (CIM) e citometria de fluxo, a atividade de oito agentes antibacterianos (enrofloxacina, ciprofloxacina, gentamicina, claritromicina, cloranfenicol, oxitetraclina, tiamulina e tilosina) contra Ureaplasma diversum. Foram analisadas 24 amostras de isolados de campo oriundas da mucosa genital de fêmeas bovinas. As amostras foram confirmadas por crescimento em caldo, placa e por PCR. Os inóculos foram submetidos à analise de suscetibilidade aos antibióticos pelo método da microdiluição em microplaca e posteriormente analisados pelo citômetro de fluxo a fim de avaliar a atividade antimicrobiana nas células. A claritromicina apresentou os maiores índices de inibição in vitro, sendo a gentamicina considerada o antibiótico de menor espectro de ação nesse estudo. De acordo com as análises do citômetro, a gentamicina apresentou o menor número de células viáveis enquanto a tiamulina apresentou o maior número. Embora haja resultados destoantes entre as técnicas utilizadas, o citômetro de fluxo pode ser utilizado como uma boa ferramenta para auxiliar a avaliação da suscetibilidade desses microrganismos a antibióticos.
The Mollicutes cause disease in several economically important species, including cattle. In this study, was evaluated by minimum inhibitory concentration (MIC) and flow cytometry, the activity of eight antibacterial agents (enrofloxacin, ciprofloxacin, gentamicin, clarithromycin, chloramphenicol, oxitetraclina, tiamulin and tylosin) against Ureaplasma diversum. We analyzed 24 samples of field isolates originating from the genital mucosa of cows. The samples were confirmed by growth in broth, plate, and PCR. The inoculations were subjected to analysis of susceptibility to antibiotics by the method of micro-dilution plate and then analyzed by flow cytometry to assess the antimicrobial activity in cells. Clarithromycin showed the highest levels of inhibition in vitro, the antibiotic gentamicin considered lower spectrum of action in this study. According to the analysis of the flow cytometer, gentamicin showed the lowest number of viable cells as tiamulin showed the greatest number. Although there are divergent results between the techniques used, flow cytometry can be used as a good tool even help assess the susceptibility of microorganisms to antibiotics.

O desenvolvimento do câncer cervical depende da exposição ao HPV, fator necessário, mas não suficiente. Outras bactérias, tais como ureaplasmas, têm sido associadas como cofatores. O objetivo deste estudo foi avaliar a presença de ureaplasmas em mulheres com lesão cervical, e observar alterações em PHK causadas pela infecção por ureaplasmas. 140 swabs vaginais foram coletados. O material foi submetido a PCR para a detecção de HPV, Mollicutes, U. urealyticum, U. parvum e seus sorotipos, e outras bactérias de importância ginecológica; e qPCR para U. urealyticum e U. parvum. Também foi realizada a infecção de ureaplasmas em PHK transformados com HPV. As células foram contadas e realizou-se a dosagem das citocinas IL1-β, IL-6 e TNF-α. HPV, Mollicutes, U. parvum, sorotipos 1 e 6 de U. parvum, T. vaginalis e G. vaginalis, além de alguns fatores socioeconômicos, foram associados com lesão cervical. Verificou-se maior carga de U. parvum entre mulheres com lesão. Houve diminuição do número de células e maior liberação de IL-6 e TNF-α nos grupos infectados. Com os resultados obtidos neste estudo, foi possível verificar uma associação entre os ureaplasmas e HPV no início das lesões cervicais, contudo mais estudos precisam ser realizados para aprimorar essa hipótese.
The development of cervical cancer depends on the exposure to HPV, necessary factor, but not enough. Other bacteria, such as ureaplasmas, have been associated as cofactors. The aim of this study was to evaluate the presence of ureaplasmas in women with cervical injury, and observe changes in PHK infected by ureaplasmas. 140 vaginal swabs were collected. The material was subjected to PCR for detection of HPV, Mollicutes, Ureaplasma urealyticum, U. parvum (and serotypes) and other bacteria gynecological importance; qPCR for U. urealyticum and U. parvum was made. PHK transformed by HPV was infected by ureaplasma. Cells were counted and it was done titration of IL1-β, IL-6 and TNF-α. HPV, Mollicutes, U. parvum, serotypes 1 and 6 U. parvum, T. vaginalis and G. vaginalis, and some socioeconomic factors were associated with cervical injury. Besides this, it was detected higher load U. parvum among women with injury. There was decrease in cell number and increased release of IL-6 and TNF-α in infected groups. With the results of this study, we found an association among HPV and ureaplasmas at the beginning of cervical lesions, but more studies are needed to enhance this hypothesis.

Monoclonal antibodies (Mabs) raised against Ureaplasma urealyticum (serotype 8) revealed the presence of three membrane antigens. One major surface antigen of apparent molecular mass 96 kDa, shown to express four distinct epitopes, was found to be serotype-8 specific. Thus, Mabs raised against this polypeptide will unequivocally differentiate serotype 8 from the other serotypes of human origin. The binding of antibodies to this polypeptide partially suppressed the growth of the organisms. Membrane expressed antigenic polypeptides of apparent molecular masses 16 kDa and 17 kDa were expressed by those serotypes belonging to the large serocluster (A), whereas the 17 kDa polypeptide only was expressed in the smaller serocluster (B). Using this Mab probe serotype 13 was placed in the larger serocluster. Thus Mabs, which recognise one or both of these polypeptides, will unequivocally differentiate the two seroclusters of this organism. The cytosolic urease from U. urealyticum, serotype 8, was purified by immuno-affinity chromatography. Two active forms of the enzyme were demonstrated by non-denaturing electrophoretic analysis and a single peak with urease activity of apparent molecular mass 190 KDa was shown by FPLC. Freezing and thawing of the purified enzyme caused a partial breakdown to inactive sub-units whereas total inactivation of the enzyme and denaturation, achieved by boiling for two minutes in the absence of any added denaturing agents, revealed three subunit polypeptides of apparent molecular masses 72, 14 and 11 KDa. Densitometry suggested that the active enzyme contains equimolar ratios of the three subunits and hence is a hexamer. The active enzyme displayed two pH optima of 6.9 and 6.15. Mabs raised against purified urease bound to both the active enzyme and to the inactive 72 kDa subunit. No evidence of antigenicity was found for the 14 and 11 KDa sub-units. These Mabs cross-reacted with ureases from all the other human serotypes. Competition assays revealed a minimum of four and possibly five distinct epitopes on the enzyme, all distinct from its active site. Ureaplasmas from 5 animal hosts were studied using the various Mabs. The 96 kDa antigen was not found in any of the non-human strains. Variations in the available epitopes on the ureases and the presence or absence of the 16/17 KDa antigens in the non-human strains allowed a putative identification of the source of the non-human ureaplasmas. Such investigations also showed that with the exception of the 96 KDa serotype 8-specific antigen, chimpanzee isolates could not be differentiated from the human ureaplasma serotypes belonging to the large serocluster. These Mabs were also used to develop fluorescent probes and other diagnostic assays which included a slide agglutination system and a sensitive urease catch assay which was also converted into a 'dip-stick' assay.

Com a crescente demanda por produtos ovinos, tem crescido o número de empresários dispostos a investir nessas atividades. Estudos sobre a importância das micoplasmoses e suas diferentes espécies já conhecidas ainda devem ser realizadas no Brasil. Foram estudadas 60 fêmeas da espécie ovina. O material clínico selecionado para analisar foi muco vulvovaginal obtido através de colheita por zaragatoa. Foi obtido o isolamento através de cultivo e a detecção pela técnica de reação em cadeia da polimerase de Mycoplasma spp., Ureaplasma spp. e Acholeplasma laidlawii em fêmeas de ovinos pela primeira vez no Brasil na região vulvovaginal. A porcentagem de micoplasma genérico detectada nas amostras foi de 75%, de Ureaplasma spp. foi de 66,7% e de Acholeplasma laidwaii foi de 1,7%. Houve isolamento em placa de 11,7% de Ureaplasma spp. do total de amostras processadas e de Micoplasma spp. em 8,3%. Não foi possível a identificação da espécie envolvida. Não houve associação da presença de micoplasmas com alterações no trato reprodutivo de fêmeas ovinas. Foi comprovada a infecção de uma fêmea ovina através de monta natural por um macho infectado, provando que uma das principais fontes de infecção dos animais é através do coito.
Sheep industry is growing rapidly in Brazil. However many diseases are the principal bottleneck to the increasing these activities. Among the reproductive diseases the vulvo-vaginites caused by Mycoplasma play an important role. Although no studies has been carried out in brazilian sheep herd so far. For that 60 ewes were ramdomly from different flocks selected at the county of Piedade state of São Paulo, Brazil. The mucus from vulvovagina was obtained by swab for isolation and detectetion of Mycoplasma spp., Ureaplasma spp. and Acholeplasma laidlawii. Generic Mycoplasma was detected in 75% of the samples, Ureaplasma spp. in 66,7% and Acholeplasma laidwaii in 1,7%. Mycoplasma spp. was isolated 8,3% and Ureaplasma spp. in 11,7%. It was not possible the identification of the species found. There was no association of mycoplasmas in the muco on the presence of reproductive problems in the herds. At least in one case, ram with presence of Mycoplasma in the semen infected a negative ewe, suggesting the importance of the ram in spreading the bacteria in the heard.

The Mycoplasma species Ureaplasma parvum and Ureaplasma urealyticum colonize the human adult urogenital tract and are not typically associated with disease. Perinatal transmission, however, has been implicated in the pathogenesis of preterm birth, chorioamnionitis, and other complications of extreme prematurity, including neonatal pneumonitis, bronchopulmonary dysplasia (BPD), meningitis, and necrotizing enterocolitis (NEC). This chapter reviews the biology of these organisms. Epidemiologic and experimental evidence supporting a role for ureaplasmas in the pathogenesis of neonatal disease, clinical manifestations of infection in the infant, current microbiologic diagnostic methods, and the present status of treatment options are reviewed. Macrolide antibiotic therapy is controversial for infected infants, and current concepts regarding candidates for treatment are discussed. Key unanswered questions that need to be addressed in future research studies are also suggested.

Introdução: A microbiota vaginal é um complexo sistema com diversidade de microrganismos. A disbiose parece aumentar o risco de infecções, principalmente as sexualmente transmissíveis, entre as quais por papilomavírus humano, agente associado a lesões cervicais. Objetivo: Avaliar os diferentes tipos de microbiota cervical e as suas características no esfregaço de material residual de citologia em meio líquido, associando com o papilomavírus humano e com Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Trichomonas vaginalis. Métodos: O estudo analisou 179 casos que tinham material residual de citologia em meio líquido. Alíquota do material foi colocado em lâmina adequada, fixado a seco e corado por método de Gram para leitura em microscópio óptico. Outra alíquota foi utilizada para estudo em reação em cadeia da polimerase - transcriptase reversa e multiplex para pesquisas dos microorganismos associados a infecções sexualmente transmissíveis. O teste exato de Fisher com intervalo de confiança foi utilizado para significância estatística. O projeto foi aprovado em comitê de ética sob número 24071519.9.0000.5049 (UniChristus). Resultados: Os casos foram divididos conforme o escore de Nugent aplicado a esfregaços corados pelo método de Gram. Em microbiota cervical normal (escores de 0 a 3), 100 casos (55,86%); em microbiota intermediária (escore de 4 a 6), 51 casos (28,5%); em sugestivo de disbiose (escore de 7 a 10), 28 casos (15,64%). Nos casos de disbiose, foram observados: Chlamydia trachomatis (1[3,57%]), Mycoplasma hominis (7[25%]), Ureaplasma urealyticum (1[3,57%]), papilomavírus humano 16/45 (1[3,57%]), papilomavírus humano de alto risco (AR) (3[10,71%]) e AR e 16/45 (1[3,57%]). No grupo normal, foi a seguinte distribuição: Ureaplasma urealyticum (1[1%]), papilomavírus humano 16 (2[2%]), papilomavírus humano 18/45 (3[3%]), AR (13[13%]). No grupo intermediário, a distribuição foi: Ureaplasma urealyticum (2[3,9%]), papilomavírus humano AR (5[9,8%]) e papilomavírus humano AR, 16 (1[3,9%]). A única diferença significativa foi de Mycoplasma hominis na disbiose (p<0,0001). Conclusão: O estudo não evidenciou uma associação maior no grupo de disbiose com a maioria das infecções sexualmente transmissíveis, no entanto, com Mycoplasma hominis, foi significativo.