RESUMO
BACKGROUND: Ureaplasma diversum is a pathogen found in the genital tract of cattle and associated with genital disorders such as infertility, placentitis, abortion, birth of weak calves, low sperm motility, seminal vesiculitis and epididymitis. There are few studies evaluating the genetic diversity of U. diversum strains and their influence on the immune response in cattle. Therefore, to better understand genetic relationships of the pathogenicity of U. diversum, a multilocus sequence typing (MLST) scheme was performed to characterize the ATCC 49782 strain and another 40 isolates recovered from different Brazilian states. RESULTS: Primers were designed for housekeeping genes ftsH, polC, rpL22, rpoB, valS and ureA and for virulence genes, phospholipase D (pld), triacylglycerol lipase (tgl), hemolysin (hlyA), MIB-MIP system (mib,mip), MBA (mba), VsA (VsA) and ribose transporter (tABC). PCRs were performed and the targeted gene products were purified and sequenced. Sequence types (STs), and clonal complexes (CCs) were assigned and the phylogenetic relationship was also evaluated. Thus, a total of 19 STs and 4 CCs were studied. Following the molecular analysis, six isolates of U. diversum were selected, inoculated into bovine monocyte/macrophage culture and evaluated for gene expression of the cytokines TNF-α, IL-1, IL-6, IL-10 and IL-17. Differences were detected in the induction of cytokines, especially between isolates 198 and BA78, promoted inflammatory and anti-inflammatory profiles, respectively, and they also differed in virulence factors. CONCLUSION: It was observed that intra-species variability between isolates of U. diversum can induce variations of virulent determinants and, consequently, modulate the expression of the triggered immune response.
Assuntos
Infecções por Ureaplasma/veterinária , Ureaplasma/genética , Ureaplasma/imunologia , Animais , Brasil , Bovinos , Doenças dos Bovinos/microbiologia , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , Feminino , Masculino , Tipagem de Sequências Multilocus/veterinária , Filogenia , Ureaplasma/classificação , Ureaplasma/patogenicidade , Infecções por Ureaplasma/imunologia , Virulência/genéticaAssuntos
Coinfecção/epidemiologia , Infecções por Mycoplasma/epidemiologia , Mycoplasma/isolamento & purificação , Papillomaviridae/isolamento & purificação , Lesões Pré-Cancerosas/epidemiologia , Infecções por Ureaplasma/epidemiologia , Ureaplasma/isolamento & purificação , Adolescente , Adulto , Idoso , Estudos Transversais , Feminino , Genótipo , Humanos , México/epidemiologia , Pessoa de Meia-Idade , Mycoplasma/classificação , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/microbiologia , Papillomaviridae/genética , Reação em Cadeia da Polimerase , Prevalência , Ureaplasma/classificação , Infecções por Ureaplasma/diagnóstico , Infecções por Ureaplasma/microbiologiaRESUMO
Ureaplasma diversum is a member of the Mollicutes class responsible for urogenital tract infection in cattle and small ruminants. Studies indicate that the process of horizontal gene transfer, the exchange of genetic material among different species, has a crucial role in mollicute evolution, affecting the group's characteristic genomic reduction process and simplification of metabolic pathways. Using bioinformatics tools and the STRING database of known and predicted protein interactions, we constructed the protein-protein interaction network of U. diversum and compared it with the networks of other members of the Mollicutes class. We also investigated horizontal gene transfer events in subnetworks of interest involved in purine and pyrimidine metabolism and urease function, chosen because of their intrinsic importance for host colonization and virulence. We identified horizontal gene transfer events among Mollicutes and from Ureaplasma to Staphylococcus aureus and Corynebacterium, bacterial groups that colonize the urogenital niche. The overall tendency of genome reduction and simplification in the Mollicutes is echoed in their protein interaction networks, which tend to be more generalized and less selective. Our data suggest that the process was permitted (or enabled) by an increase in host dependence and the available gene repertoire in the urogenital tract shared via horizontal gene transfer.
Assuntos
Proteínas de Bactérias/metabolismo , Transferência Genética Horizontal , Genoma Bacteriano , Mapas de Interação de Proteínas , Tenericutes/genética , Ureaplasma/genética , Animais , Proteínas de Bactérias/genética , Bovinos , Corynebacterium/genética , Evolução Molecular , Tamanho do Genoma , Genômica , Redes e Vias Metabólicas , Purinas/metabolismo , Pirimidinas/metabolismo , Staphylococcus aureus/genética , Tenericutes/classificação , Tenericutes/metabolismo , Ureaplasma/classificação , Ureaplasma/metabolismo , VirulênciaRESUMO
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.
Assuntos
Muco do Colo Uterino/microbiologia , Infecções por Ureaplasma/veterinária , Ureaplasma/isolamento & purificação , Vagina/microbiologia , Criação de Animais Domésticos , Animais , Bovinos , Fazendas , Feminino , Poaceae/microbiologia , Reação em Cadeia da Polimerase , Análise de Regressão , Fatores de Risco , Ureaplasma/classificação , Infecções por Ureaplasma/microbiologiaRESUMO
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.
Assuntos
Animais , Feminino , Ureaplasma/isolamento & purificação , Vagina/microbiologia , Muco do Colo Uterino/microbiologia , Infecções por Ureaplasma/veterinária , Ureaplasma/classificação , Bovinos , Reação em Cadeia da Polimerase , Análise de Regressão , Fatores de Risco , Infecções por Ureaplasma/microbiologia , Fazendas , Poaceae/microbiologia , Criação de Animais DomésticosRESUMO
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.
Assuntos
Animais , Masculino , Feminino , Sêmen/microbiologia , Doenças dos Ovinos/microbiologia , Ureaplasma/isolamento & purificação , Vagina/microbiologia , Doenças das Cabras/microbiologia , Infecções por Ureaplasma/veterinária , Ureaplasma/classificação , Ureaplasma/genética , Brasil , Cabras , Ovinos , Infecções por Ureaplasma/microbiologiaRESUMO
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.
Assuntos
Doenças das Cabras/microbiologia , Sêmen/microbiologia , Doenças dos Ovinos/microbiologia , Infecções por Ureaplasma/veterinária , Ureaplasma/isolamento & purificação , Vagina/microbiologia , Animais , Brasil , Feminino , Cabras , Masculino , Ovinos , Ureaplasma/classificação , Ureaplasma/genética , Infecções por Ureaplasma/microbiologiaAssuntos
Pneumonia/microbiologia , Pneumonia/veterinária , Doenças dos Suínos/microbiologia , Infecções por Ureaplasma/veterinária , Ureaplasma/isolamento & purificação , Animais , Cuba , DNA Bacteriano , Dados de Sequência Molecular , Filogenia , Suínos , Ureaplasma/classificação , Infecções por Ureaplasma/microbiologiaRESUMO
Ureaplasma diversum has been associated with reproductive disorders in cattle and in the present study genotypic variations among U. diversum isolates obtained from the vaginal mucus of healthy cattle and sick animals were analyzed by enzymatic digestion and pulsed-field gel electrophoresis (PFGE). The influence of time and broth volume was important in obtaining sufficient cell sediment and DNA for PFGE. The method presented a high discriminatory power and satisfactory reproducibility for the analysis of detected variations among U. diversum isolates and strains. Different band profiles and wide genotypic heterogeneity were detected but no association between DNA polymorphism and sick or healthy animals could be established.
Assuntos
Doenças dos Bovinos/microbiologia , DNA Bacteriano/análise , Infecções por Ureaplasma/veterinária , Ureaplasma/genética , Animais , Bovinos , Eletroforese em Gel de Campo Pulsado/veterinária , Feminino , Genótipo , Polimorfismo Genético , Ureaplasma/classificação , Infecções por Ureaplasma/microbiologiaRESUMO
Ureaplasma parvum and Ureaplasma urealyticum, also known as biovar parvum and biovar T960, respectively, could be associated with several disorders in men, women, and mainly, in newborn children with under weight. Several methods have been developed in order to identify the species or biovars of ureaplasmas. We developed a Multiplex-PCR method using the UPS-UPSA and UUS2-UUA2 primers, specific for U. parvum and U. urealyticum, respectively. This Multiplex-PCR method was used to identify cultures of clinical positive samples to Ureaplasma spp. by the "MYCOFAST Evolution-2" Kit. Of 56 positive cultures to Ureaplasma spp. from newborn children, 70% were U. parvum and 30% U. urealyticum; in 76 positive samples in women, 83% corresponded to U. parvum and 17% to U. urealyticum, while in 63 positive samples of men, 76% identified U. parvum and 24% U. urealyticum. The PCR-multiplex method showed specificity for the identification of the biovars or species of ureaplasmas of clinical interest.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Reação em Cadeia da Polimerase/métodos , Ureaplasma urealyticum/classificação , Ureaplasma/classificação , Adulto , Primers do DNA , DNA Bacteriano/isolamento & purificação , Feminino , Humanos , Recém-Nascido , Masculino , Especificidade da Espécie , Ureaplasma/genética , Ureaplasma/isolamento & purificação , Infecções por Ureaplasma/microbiologia , Ureaplasma urealyticum/genética , Ureaplasma urealyticum/isolamento & purificaçãoRESUMO
Los micoplasmas revisten gran importancia clínica, por haberse reconocido su papel patógeno en entidades clínicas como neumonía atípica primaria, uretritis no gonocócica e infecciones cervicovaginales de la mujer en etapa sexual activa. En Europa durante el siglo XVIII, se presentó en el ganado una enfermedad pulmonar altamente contagiosa, a la cual se denominó pleuroneumonía. Sin embargo, fue hasta 1898 cuando Nocard y Roux lograron aislar el agente causal de esta enfermedad sin saber que se trataba de Mycoplasma mycoides. Las colonias poducidas en el medio de cultivo, se apreciaban muy pequeñas, pleomórficas y se teñían débilmente con los colorantes habituales. Con el paso del tiempo se siguieron observando algunos gérmenes, cuya morfología y propiedades de cultivo eran semejantes. Las fuentes de estos aislamientos fueron muy diversas. Al principio y de manera abreviada se les denominó PPLO (Pleuropneumoniae like organisms) por su similitud con agentes productores de pleuroneumonía bovina