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Jorge Timenetsky - One of the best experts on this subject based on the ideXlab platform.
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A Review of Ureaplasma diversum: A Representative of the Mollicute Class Associated With Reproductive and Respiratory Disorders in Cattle
Frontiers in Veterinary Science, 2021Co-Authors: Manoel Neres Santos Junior, Jorge Timenetsky, Guilherme Barreto Campos, Nayara Silva De Macêdo Neres, Bruno Lopes Bastos, Lucas Miranda MarquesAbstract:The Mollicutes class encompasses wall-less microbes with a reduced genome. They may infect plants, insects, humans, and animals including those on farms and in livestock. Ureaplasma diversum is a mollicute associated with decreased reproduction mainly in the conception rate in cattle, as well as weight loss and decreased quality in milk production. Therefore, U. diversum infection contributes to important economic losses, mainly in large cattle-producing countries such as the United States, China, Brazil, and India. The characteristics of Mollicutes, virulence, and pathogenic variations make it difficult to control their infections. Genomic analysis, prevalence studies, and immunomodulation assays help better understand the pathogenesis of bovine Ureaplasma. Here we present the main features of transmission, virulence, immune response, and pathogenesis of U. diversum in bovines.
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Multilocus sequence typing characterizes diversity of Ureaplasma diversum strains, and intra-species variability induces different immune response profiles
BMC Veterinary Research, 2020Co-Authors: Yasmin M. F. S. Andrade, Izadora S. Rezende, Guilherme B. Campos, Aline T. Amorim, Manoel N. Santos-junior, Maysa S. Barbosa, Ícaro B. S. Silva, Ellunny C. Queiroz, Bruno L. Bastos, Jorge TimenetskyAbstract: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.
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Ureaplasma diversum protein interaction networks: evidence of horizontal gene transfer and evolution of reduced genomes among Mollicutes
Canadian journal of microbiology, 2019Co-Authors: Joana Kästle Silva, Lucas M. Marques, Jorge Timenetsky, Sávio Torres De FariasAbstract: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.
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Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice
Anais da Academia Brasileira de Ciências, 2016Co-Authors: Jamile R. Silva, Lucas M. Marques, Lício F.a.a. Ferreira, Percíllia V.s. Oliveira, Ivanéia V. Nunes, Ítalo S. Pereira, Jorge Timenetsky, Tiana B. Figueiredo, Robson A.a. SilvaAbstract:Ureaplasma diversum is an opportunistic pathogen associated with uterine inflammation, impaired embryo implantation, infertility, abortions, premature birth of calves and neonatal pneumonia in cattle. It has been suggested that the intra-uterine infection by Ureaplasma diversum can cause vascular changes that hinder the success of pregnancy. Thus, the aim of this study was to evaluate the changes of intrauterine site of A/J mice in estrus or proestrus phase inoculated with Ureaplasma diversum. The infection was monitored at 24, 48 and 72 hours by the PCR methodology to detect the Ureaplasma in the inoculation site and the profile of circulating blood cells. Morphological changes, intensity of inflammation and the production of cytokines were compared. The infected mice showed local inflammation through the production of IFN-γ and TNF-α. Ureaplasma diversum infections in the reproductive tract of studied mice seemed to be associated with the production of pro-inflammatory cytokines in uterine parenchyma. The levels of TNF-α of infected mice were dependent on the bacterial load of inoculated Ureaplasma. Uterine experimental infections by Ureaplasma diversum have not been mentioned yet and herein we presented the first report of an intrauterine infection model in mice.
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A quantitative TaqMan PCR assay for the detection of Ureaplasma diversum.
Veterinary microbiology, 2013Co-Authors: Lucas M. Marques, Maysa S. Barbosa, Hellen B. Martins, Guilherme B. Campos, Aline T. Amorim, Izadora Souza Rezende, Tassia Neves Lobao, Jorge TimenetskyAbstract:Ureaplasma diversum in veterinary studies is an undesirable microbe, which may cause infection in bulls and may result in seminal vesiculitis, balanopostitis, and alterations in spermatozoids, whereas in cows, it may cause placentitis, fetal alveolitis, abortion, and birth of weak calves. U. diversum is released through organic secretions, especially semen, preputial and vaginal mucus, conjunctival secretion, and milk. The aim of the present study was to develop a TaqMan probe, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of U. diversum from genital swabs of bovines. Primers and probes specific to U. diversum 16S rRNA gene were designed. The specificity, detection limit, intra- and inter-assay variability of qPCR to detect this Ureaplasma was compared with the results of the conventional PCR assay (cPCR). Swabs of vaginal mucus from 169 cows were tested. The qPCR assay detected as few as 10 copies of U. diversum and was 100-fold more sensitive than the cPCR. No cross-reactivity with other Mollicutes or eubacteria was observed. U. diversum was detected in 79 swabs (46.42%) by qPCR, while using cPCR it was detected in 42 (25%) samples. The difference in cPCR and qPCR Ureaplasma detection between healthy and sick animals was not statistically significant. But the U. diversum load in samples from animals with genital disorders was higher than in healthy animals. The qPCR assay developed herein is highly sensitive and specific for the detection and quantification of U. diversum in vaginal bovine samples.
Melissa Buzinhani - One of the best experts on this subject based on the ideXlab platform.
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SHORT REPORT Open Access Invasion of Ureaplasma diversum in bovine spermatozoids
2013Co-Authors: Melissa Buzinhani, Mauricio Yamaguti, Rosangela C Oliveira, Beatriz Araujo Cortez, Mayra Eo Assumpcao, Gláucia M Machado-santelli, Lucas Mir, A Marques, Jorge TimenetskyAbstract:Background: Ureaplasma diversum has been associated with infertility in cows. In bulls, this mollicute colonizes the prepuce and distal portion of the urethra and may infect sperm cells. The aim of this study is to analyze in vitro interaction of U. diversum isolates and ATCC strains with bovine spermatozoids. The interactions were observed by confocal microscopy and the gentamycin internalization assay. Findings: U. diversum were able to adhere to and invade spermatozoids after 30 min of infection. The gentamicin resistance assay confirmed the intracellularity and survival of U. diversum in bovine spermatozoids. Conclusions: The intracellular nature of bovine Ureaplasma identifies a new difficulty to control the reproductive of these animals
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invasion of Ureaplasma diversum in bovine spermatozoids
BMC Research Notes, 2011Co-Authors: Melissa Buzinhani, Mauricio Yamaguti, Rosangela C Oliveira, Beatriz Araujo Cortez, Lucas Miranda Marques, Glaucia Maria Machadosantelli, Mayra Eo Assumpcao, Jorge TimenetskyAbstract:Background Ureaplasma diversum has been associated with infertility in cows. In bulls, this mollicute colonizes the prepuce and distal portion of the urethra and may infect sperm cells. The aim of this study is to analyze in vitro interaction of U. diversum isolates and ATCC strains with bovine spermatozoids. The interactions were observed by confocal microscopy and the gentamycin internalization assay.
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Invasion of Ureaplasma diversum in bovine spermatozoids
BMC Research Notes, 2011Co-Authors: Melissa Buzinhani, Mauricio Yamaguti, Rosangela C Oliveira, Beatriz Araujo Cortez, Lucas Miranda Marques, Mayra Eo Assumpcao, Gláucia M Machado-santelli, Jorge TimenetskyAbstract:Background Ureaplasma diversum has been associated with infertility in cows. In bulls, this mollicute colonizes the prepuce and distal portion of the urethra and may infect sperm cells. The aim of this study is to analyze in vitro interaction of U. diversum isolates and ATCC strains with bovine spermatozoids. The interactions were observed by confocal microscopy and the gentamycin internalization assay. Findings U. diversum were able to adhere to and invade spermatozoids after 30 min of infection. The gentamicin resistance assay confirmed the intracellularity and survival of U. diversum in bovine spermatozoids. Conclusions The intracellular nature of bovine Ureaplasma identifies a new difficulty to control the reproductive of these animals.
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Intraspecific sequence variation in 16S rRNA gene of Ureaplasma diversum isolates
Veterinary Microbiology, 2011Co-Authors: Lucas M. Marques, Ana M. S. Guimarães, Melissa Buzinhani, Mauricio Yamaguti, Rosangela C Oliveira, R L Neto, R.c.p. Marques, Sávio Torres De Farias, Jorge TimenetskyAbstract:Ureaplasma diversum infection in bulls may result in seminal vesiculitis, balanoposthitis and alterations in spermatozoids. In cows, it can cause placentitis, fetal alveolitis, abortion and the birth of weak calves. U. diversum ATCC 49782 (serogroups A), ATCC 49783 (serogroup C) and 34 field isolates were used for this study. These microorganisms were submitted to Polymerase Chain Reaction for 16S gene sequence determination using Taq High Fidelity and the products were purified and bi-directionally sequenced. Using the sequence obtained, a fragment containing four hypervariable regions was selected and nucleotide polymorphisms were identified based on their position within the 16S rRNA gene. Forty-four single nucleotide polymorphisms (SNP) were detected. The genotypic variability of the 16S rRNA gene of U. diversum isolates shows that the taxonomy classification of these organisms is likely much more complex than previously described and that 16S rRNA gene sequencing may be used to suggest an epidemiologic pattern of different origin strains.
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Invasion of Ureaplasma diversum in Hep-2 cells
BMC Microbiology, 2010Co-Authors: Lucas Miranda Marques, Melissa Buzinhani, Mauricio Yamaguti, Rosangela C Oliveira, Beatriz Araujo Cortez, Priscilla M Ueno, Renata L Neto, Ana Márcia S Guimarães, Telma A Monezi, Antonio Carlos R BragaAbstract:Background Understanding mollicutes is challenging due to their variety and relationship with host cells. Invasion has explained issues related to their opportunistic role. Few studies have been done on the Ureaplasma diversum mollicute, which is detected in healthy or diseased bovine. The invasion in Hep-2 cells of four clinical isolates and two reference strains of their Ureaplasma was studied by Confocal Laser Scanning Microscopy and gentamicin invasion assay. Results The isolates and strains used were detected inside the cells after infection of one minute without difference in the arrangement for adhesion and invasion. The adhesion was scattered throughout the cells, and after three hours, the invasion of the Ureaplasmas surrounded the nuclear region but were not observed inside the nuclei. The gentamicin invasion assay detected that 1% of the ATCC strains were inside the infected Hep-2 cells in contrast to 10% to the clinical isolates. A high level of phospholipase C activity was also detected in all studied Ureaplasma. Conclusions The results presented herein will help better understand U. diversum infections, aswell as cellular attachment and virulence.
Lucas M. Marques - One of the best experts on this subject based on the ideXlab platform.
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Ureaplasma diversum protein interaction networks: evidence of horizontal gene transfer and evolution of reduced genomes among Mollicutes
Canadian journal of microbiology, 2019Co-Authors: Joana Kästle Silva, Lucas M. Marques, Jorge Timenetsky, Sávio Torres De FariasAbstract: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.
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Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice
Anais da Academia Brasileira de Ciências, 2016Co-Authors: Jamile R. Silva, Lucas M. Marques, Lício F.a.a. Ferreira, Percíllia V.s. Oliveira, Ivanéia V. Nunes, Ítalo S. Pereira, Jorge Timenetsky, Tiana B. Figueiredo, Robson A.a. SilvaAbstract:Ureaplasma diversum is an opportunistic pathogen associated with uterine inflammation, impaired embryo implantation, infertility, abortions, premature birth of calves and neonatal pneumonia in cattle. It has been suggested that the intra-uterine infection by Ureaplasma diversum can cause vascular changes that hinder the success of pregnancy. Thus, the aim of this study was to evaluate the changes of intrauterine site of A/J mice in estrus or proestrus phase inoculated with Ureaplasma diversum. The infection was monitored at 24, 48 and 72 hours by the PCR methodology to detect the Ureaplasma in the inoculation site and the profile of circulating blood cells. Morphological changes, intensity of inflammation and the production of cytokines were compared. The infected mice showed local inflammation through the production of IFN-γ and TNF-α. Ureaplasma diversum infections in the reproductive tract of studied mice seemed to be associated with the production of pro-inflammatory cytokines in uterine parenchyma. The levels of TNF-α of infected mice were dependent on the bacterial load of inoculated Ureaplasma. Uterine experimental infections by Ureaplasma diversum have not been mentioned yet and herein we presented the first report of an intrauterine infection model in mice.
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General features of the genome of Ureaplasma diversum ATCC 49782 compared to human Ureaplasmas and other members of Mycoplasma, Acholeplasma and Phytoplasma species.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Verena M. SantosAbstract:General features of the genome of Ureaplasma diversum ATCC 49782 compared to human Ureaplasmas and other members of Mycoplasma, Acholeplasma and Phytoplasma species.
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Coding DNA sequences (CDSs) of Ureaplasma diversum ATCC 49782 genome classified by TIGR role category.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Verena M. SantosAbstract:Coding DNA sequences (CDSs) of Ureaplasma diversum ATCC 49782 genome classified by TIGR role category.
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Virulence and pathogenicity mechanisms.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Verena M. SantosAbstract:(A) Virulence map of U. diversum ATCC 49782. (B) Schematic representation of the urease gene cluster from U. diversum ATCC 49782. Structural subunits: ureA (gudiv_255), ureB (gudiv_254), and ureC (gudiv_253). Accessory proteins ureE (gudiv_252), ureF (gudiv_251), ureG (gudiv_250), and ureD (gudiv_249) (C) Diagram of Ureaplasma diversum Multiple-Banded Antigen-like protein (MBA-like—gudiv_653) and locus and similarity of MBA-like with the human Ureaplasmal Multiple-Banded Antigen (MBA) (Accession number: AF055358.2).
Aline T. Amorim - One of the best experts on this subject based on the ideXlab platform.
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Multilocus sequence typing characterizes diversity of Ureaplasma diversum strains, and intra-species variability induces different immune response profiles
BMC Veterinary Research, 2020Co-Authors: Yasmin M. F. S. Andrade, Izadora S. Rezende, Guilherme B. Campos, Aline T. Amorim, Manoel N. Santos-junior, Maysa S. Barbosa, Ícaro B. S. Silva, Ellunny C. Queiroz, Bruno L. Bastos, Jorge TimenetskyAbstract: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.
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Ureaplasma diversum Genome Provides New Insights about the Interaction of the Surface Molecules of This Bacterium with the Host
PloS one, 2016Co-Authors: Lucas Miranda Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Aline T. Amorim, Naíla C. Do Nascimento, Andrea Pires Dos Santos, Verena M. SantosAbstract:Whole genome sequencing and analyses of Ureaplasma diversum ATCC 49782 was undertaken as a step towards understanding U. diversum biology and pathogenicity. The complete genome showed 973,501 bp in a single circular chromosome, with 28.2% of G+C content. A total of 782 coding DNA sequences (CDSs), and 6 rRNA and 32 tRNA genes were predicted and annotated. The metabolic pathways are identical to other human Ureaplasmas, including the production of ATP via hydrolysis of the urea. Genes related to pathogenicity, such as urease, phospholipase, hemolysin, and a Mycoplasma Ig binding protein (MIB)-Mycoplasma Ig protease (MIP) system were identified. More interestingly, a large number of genes (n = 40) encoding surface molecules were annotated in the genome (lipoproteins, multiple-banded antigen like protein, membrane nuclease lipoprotein and variable surface antigens lipoprotein). In addition, a gene encoding glycosyltransferase was also found. This enzyme has been associated with the production of capsule in mycoplasmas and Ureaplasma. We then sought to detect the presence of a capsule in this organism. A polysaccharide capsule from 11 to 17 nm of U. diversum was observed trough electron microscopy and using specific dyes. This structure contained arabinose, xylose, mannose, galactose and glucose. In order to understand the inflammatory response against these surface molecules, we evaluated the response of murine macrophages J774 against viable and non-viable U. diversum. As with viable bacteria, non-viable bacteria were capable of promoting a significant inflammatory response by activation of Toll like receptor 2 (TLR2), indicating that surface molecules are important for the activation of inflammatory response. Furthermore, a cascade of genes related to the inflammasome pathway of macrophages was also up-regulated during infection with viable organisms when compared to non-infected cells. In conclusion, U. diversum has a typical Ureaplasma genome and metabolism, and its surface molecules, including the identified capsular material, represent major components of the organism immunopathogenesis.
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General features of the genome of Ureaplasma diversum ATCC 49782 compared to human Ureaplasmas and other members of Mycoplasma, Acholeplasma and Phytoplasma species.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Verena M. SantosAbstract:General features of the genome of Ureaplasma diversum ATCC 49782 compared to human Ureaplasmas and other members of Mycoplasma, Acholeplasma and Phytoplasma species.
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Coding DNA sequences (CDSs) of Ureaplasma diversum ATCC 49782 genome classified by TIGR role category.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Verena M. SantosAbstract:Coding DNA sequences (CDSs) of Ureaplasma diversum ATCC 49782 genome classified by TIGR role category.
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Virulence and pathogenicity mechanisms.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Verena M. SantosAbstract:(A) Virulence map of U. diversum ATCC 49782. (B) Schematic representation of the urease gene cluster from U. diversum ATCC 49782. Structural subunits: ureA (gudiv_255), ureB (gudiv_254), and ureC (gudiv_253). Accessory proteins ureE (gudiv_252), ureF (gudiv_251), ureG (gudiv_250), and ureD (gudiv_249) (C) Diagram of Ureaplasma diversum Multiple-Banded Antigen-like protein (MBA-like—gudiv_653) and locus and similarity of MBA-like with the human Ureaplasmal Multiple-Banded Antigen (MBA) (Accession number: AF055358.2).
Maysa S. Barbosa - One of the best experts on this subject based on the ideXlab platform.
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Immunoinformatics and analysis of antigen distribution of Ureaplasma diversum strains isolated from different Brazilian states
BMC Veterinary Research, 2020Co-Authors: Manoel Neres Santos Junior, Maysa S. Barbosa, Izadora Souza Rezende, Ronaldo Silva Santos, Wanderson Souza Neves, Janaina Marinho Fernandes, Bruna Carolina Brito Guimarães, Lucas Santana Coelho Silva, Camila Pacheco Gomes, Caline Novaes Teixeira OliveiraAbstract:Background Ureaplasma diversum has numerous virulence factors that contribute to pathogenesis in cattle, including Lipid-associated membrane proteins (LAMPs). Therefore, the objectives of this study were to evaluate in silico important characteristics for immunobiological applications and for heterologous expression of 36 LAMPs of U. diversum (UdLAMPs) and, also, to verify by conventional PCR the distribution of these antigens in strains of Brazilian states (Bahia, Minas Gerais, São Paulo, and Mato Grosso do Sul). The Manatee database was used to obtain the gene and peptide sequences of the antigens. Similarity and identity studies were performed using BLASTp and direct antigenicity was evaluated by the VaxiJen v2.0 server. Epitope prediction for B lymphocytes was performed on the BepiPred v2.0 and CBTOPE v1.0 servers. NetBoLApan v1.0 was used to predict CD8^+ T lymphocyte epitopes. Subcellular location and presence of transmembrane regions were verified by the software PSORTb v3.0.2 and TMHMM v2.2 respectively. SignalP v5.0, SecretomeP v2.0, and DOLOP servers were used to predict the extracellular excretion signal. Physico-chemical properties were evaluated by the web-software ProtParam, Solpro, and Protein-sol . Results In silico analysis revealed that many UdLAMPs have desirable properties for immunobiological applications and heterologous expression. The proteins gudiv_61, gudiv_103, gudiv_517, and gudiv_681 were most promising. Strains from the 4 states were PCR positive for antigens predicted with immunogenic and/or with good characteristics for expression in a heterologous system. Conclusion These works contribute to a better understanding of the immunobiological properties of the UdLAMPs and provide a profile of the distribution of these antigens in different Brazilian states.
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Ureaplasma diversum Genome Provides New Insights about the Interaction of the Surface Molecules of This Bacterium with the Host
PloS one, 2016Co-Authors: Lucas Miranda Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Aline T. Amorim, Naíla C. Do Nascimento, Andrea Pires Dos Santos, Verena M. SantosAbstract:Whole genome sequencing and analyses of Ureaplasma diversum ATCC 49782 was undertaken as a step towards understanding U. diversum biology and pathogenicity. The complete genome showed 973,501 bp in a single circular chromosome, with 28.2% of G+C content. A total of 782 coding DNA sequences (CDSs), and 6 rRNA and 32 tRNA genes were predicted and annotated. The metabolic pathways are identical to other human Ureaplasmas, including the production of ATP via hydrolysis of the urea. Genes related to pathogenicity, such as urease, phospholipase, hemolysin, and a Mycoplasma Ig binding protein (MIB)-Mycoplasma Ig protease (MIP) system were identified. More interestingly, a large number of genes (n = 40) encoding surface molecules were annotated in the genome (lipoproteins, multiple-banded antigen like protein, membrane nuclease lipoprotein and variable surface antigens lipoprotein). In addition, a gene encoding glycosyltransferase was also found. This enzyme has been associated with the production of capsule in mycoplasmas and Ureaplasma. We then sought to detect the presence of a capsule in this organism. A polysaccharide capsule from 11 to 17 nm of U. diversum was observed trough electron microscopy and using specific dyes. This structure contained arabinose, xylose, mannose, galactose and glucose. In order to understand the inflammatory response against these surface molecules, we evaluated the response of murine macrophages J774 against viable and non-viable U. diversum. As with viable bacteria, non-viable bacteria were capable of promoting a significant inflammatory response by activation of Toll like receptor 2 (TLR2), indicating that surface molecules are important for the activation of inflammatory response. Furthermore, a cascade of genes related to the inflammasome pathway of macrophages was also up-regulated during infection with viable organisms when compared to non-infected cells. In conclusion, U. diversum has a typical Ureaplasma genome and metabolism, and its surface molecules, including the identified capsular material, represent major components of the organism immunopathogenesis.
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General features of the genome of Ureaplasma diversum ATCC 49782 compared to human Ureaplasmas and other members of Mycoplasma, Acholeplasma and Phytoplasma species.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Verena M. SantosAbstract:General features of the genome of Ureaplasma diversum ATCC 49782 compared to human Ureaplasmas and other members of Mycoplasma, Acholeplasma and Phytoplasma species.
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Coding DNA sequences (CDSs) of Ureaplasma diversum ATCC 49782 genome classified by TIGR role category.
2016Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Naíla Do C. Nascimento, Andrea Dos P. Santos, Aline T. Amorim, Verena M. SantosAbstract:Coding DNA sequences (CDSs) of Ureaplasma diversum ATCC 49782 genome classified by TIGR role category.
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Genome Sequence of Ureaplasma diversum Strain ATCC 49782
Genome announcements, 2015Co-Authors: Lucas M. Marques, Izadora S. Rezende, Maysa S. Barbosa, Ana M. S. Guimarães, Hellen B. Martins, Guilherme B. Campos, Aline T. Amorim, Naíla C. Do Nascimento, Andrea Pires Dos Santos, Verena M. SantosAbstract:Here, we report the complete genome sequence of Ureaplasma diversum strain ATCC 49782. This species is of bovine origin, having an association with reproductive disorders in cattle, including placentitis, fetal alveolitis, abortion, and birth of weak calves. It has a small circular chromosome of 975,425 bp.
