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1.
J Infect Dis ; 220(12): 1999-2008, 2019 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-31420650

RESUMO

Mycoplasma lipoproteins play a relevant role in pathogenicity and directly interact with the host immune system. Among human mycoplasmas, Mycoplasma hominis is described as a commensal bacterium that can be associated with a number of genital and extragenital conditions. Mechanisms of M. hominis pathogenicity are still largely obscure, and only a limited number of proteins have been associated with virulence. The current study focused on investigating the role of MHO_0730 as a virulence factor and demonstrated that MHO_0730 is a surface lipoprotein, potentially expressed in vivo during natural infection, acting both as a nuclease with its amino acidic portion and as a potent inducer of Neutrophil extracellular trapsosis with its N-terminal lipid moiety. Evidence for M. hominis neutrophil extracellular trap escape is also presented. Results highlight the relevance of MHO_0730 in promoting infection and modulation and evasion of innate immunity and provide additional knowledge on M. hominis virulence and survival in the host.


Assuntos
Proteínas de Bactérias/metabolismo , Armadilhas Extracelulares/imunologia , Armadilhas Extracelulares/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Infecções por Mycoplasma/imunologia , Infecções por Mycoplasma/microbiologia , Mycoplasma hominis/fisiologia , Humanos , Lipoproteínas/metabolismo , Infecções por Mycoplasma/metabolismo , Mycoplasma hominis/enzimologia , Neutrófilos/imunologia , Neutrófilos/metabolismo , Transporte Proteico , Proteínas Recombinantes , Virulência
2.
Parasitology ; 146(9): 1150-1155, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30616707

RESUMO

Trichomonas vaginalis is an anaerobic protist, responsible for the most prevalent non-viral sexually transmitted infection in humans. One of the most intriguing aspects of T. vaginalis pathobiology is the complex relationship with intracellular microbial symbionts: a group of dsRNA viruses belonging to family of Totiviridae (T. vaginalis virus), and eubacteria belonging to the Mycoplasma genus, in particular Mycoplasma hominis. Both microorganisms seem to strongly influence the lifestyle of T. vaginalis, suggesting a role of the symbiosis in the high variability of clinical presentation and sequelae during trichomoniasis. In the last few years many aspects of this unique symbiotic relationship have been investigated: M. hominis resides and replicates in the protozoan cell, and T. vaginalis is able to pass the bacterial infection to both mycoplasma-free protozoan isolates and human epithelial cells; M. hominis synergistically upregulates the proinflammatory response of human monocytes to T. vaginalis. Furthermore, the influence of M. hominis over T. vaginalis metabolism and physiology has been characterized. The identification of a novel species belonging to the class of Mollicutes (Candidatus Mycoplasma girerdii) exclusively associated to T. vaginalis opens new perspectives in the research of the complex series of events taking place in the multifaceted world of the vaginal microbiota, both under normal and pathological conditions.


Assuntos
Infecções por Mycoplasma/microbiologia , Mycoplasma hominis/fisiologia , Simbiose , Vaginite por Trichomonas/parasitologia , Trichomonas vaginalis/patogenicidade , Feminino , Humanos , Inflamação , Microbiota , Mycoplasma hominis/imunologia , Infecções Sexualmente Transmissíveis/imunologia , Infecções Sexualmente Transmissíveis/parasitologia , Totiviridae/metabolismo , Trichomonas vaginalis/imunologia , Vagina/microbiologia , Vagina/parasitologia
3.
Parasitol Res ; 118(12): 3565-3570, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31701295

RESUMO

The flagellated protozoon Trichomonas vaginalis, responsible for trichomoniasis, can establish a symbiotic relationship with the bacterium Mycoplasma hominis and can harbor double-stranded RNA Trichomonasvirus (TVV). In this study, we investigated by real-time PCR the prevalence of the four TVVs and of M. hominis among 48 T. vaginalis strains isolated in Italy, and we evaluated a possible association with metronidazole resistance. Fifty percent of the analyzed trichomonad strains tested positive for at least one TVV T. vaginalis, with TVV2 being the most prevalent, followed by TVV1 and TVV3. Two T. vaginalis strains were infected by TVV4, detected in Europe for the first time. Interestingly, we found more than one TVV species in 75% of positive trichomonad strains. M. hominis was present in 81.25% of T. vaginalis isolates tested, and no statistically significant association was observed with the infection by any TVV. Metronidazole sensitivity of T. vaginalis isolates was evaluated in vitro, and no correlation was observed between minimal lethal concentration and the presence of TVVs. This is the first report on TVV infection of T. vaginalis in Italy. Even if no association of TVV positive isolates with the presence of the symbiont M. hominis or with metronidazole resistance was observed, further studies are needed to shed light on the effective role of infecting microorganisms on the pathophysiology of T. vaginalis.


Assuntos
Mycoplasma hominis/isolamento & purificação , Vírus de RNA/isolamento & purificação , Trichomonas vaginalis/microbiologia , Trichomonas vaginalis/virologia , Antiprotozoários/farmacologia , Resistência a Medicamentos , Humanos , Itália , Metronidazol/farmacologia , Mycoplasma hominis/classificação , Mycoplasma hominis/genética , Mycoplasma hominis/fisiologia , Prevalência , Vírus de RNA/classificação , Vírus de RNA/genética , Vírus de RNA/fisiologia , RNA de Cadeia Dupla/genética , RNA Viral/genética , Simbiose , Tricomoníase/parasitologia , Trichomonas vaginalis/efeitos dos fármacos , Trichomonas vaginalis/fisiologia
4.
Proc Natl Acad Sci U S A ; 111(22): 8179-84, 2014 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-24843155

RESUMO

The human-infective parasite Trichomonas vaginalis causes the most prevalent nonviral sexually transmitted infection worldwide. Infections in men may result in colonization of the prostate and are correlated with increased risk of aggressive prostate cancer. We have found that T. vaginalis secretes a protein, T. vaginalis macrophage migration inhibitory factor (TvMIF), that is 47% similar to human macrophage migration inhibitory factor (HuMIF), a proinflammatory cytokine. Because HuMIF is reported to be elevated in prostate cancer and inflammation plays an important role in the initiation and progression of cancers, we have explored a role for TvMIF in prostate cancer. Here, we show that TvMIF has tautomerase activity, inhibits macrophage migration, and is proinflammatory. We also demonstrate that TvMIF binds the human CD74 MIF receptor with high affinity, comparable to that of HuMIF, which triggers activation of ERK, Akt, and Bcl-2-associated death promoter phosphorylation at a physiologically relevant concentration (1 ng/mL, 80 pM). TvMIF increases the in vitro growth and invasion through Matrigel of benign and prostate cancer cells. Sera from patients infected with T. vaginalis are reactive to TvMIF, especially in males. The presence of anti-TvMIF antibodies indicates that TvMIF is released by the parasite and elicits host immune responses during infection. Together, these data indicate that chronic T. vaginalis infections may result in TvMIF-driven inflammation and cell proliferation, thus triggering pathways that contribute to the promotion and progression of prostate cancer.


Assuntos
Macrófagos/imunologia , Neoplasias da Próstata/imunologia , Neoplasias da Próstata/parasitologia , Proteínas de Protozoários/imunologia , Tricomoníase/imunologia , Trichomonas vaginalis/imunologia , Sequência de Aminoácidos , Linhagem Celular Tumoral , Células Cultivadas , Sequência Conservada , Humanos , Oxirredutases Intramoleculares/genética , Oxirredutases Intramoleculares/imunologia , Sistema de Sinalização das MAP Quinases/imunologia , Fatores Inibidores da Migração de Macrófagos/genética , Fatores Inibidores da Migração de Macrófagos/imunologia , Macrófagos/citologia , Macrófagos/parasitologia , Masculino , Dados de Sequência Molecular , Próstata/imunologia , Próstata/parasitologia , Próstata/patologia , Neoplasias da Próstata/patologia , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Homologia de Sequência , Tricomoníase/complicações , Tricomoníase/parasitologia , Trichomonas vaginalis/genética , Trichomonas vaginalis/metabolismo
5.
Sex Transm Infect ; 91(8): 561-3, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25887282

RESUMO

OBJECTIVES: Persistence of antibodies against pathogens after antimicrobial treatment is a marker of therapy failure or evolution to a chronic infection. The kinetics of antibody production decrease following antigen elimination is highly variable, and predicting the duration of soluble immunity in infectious diseases is often impossible. This hampers the development and use of immunoassays for diagnostic and seroepidemiological purposes. In the case of Trichomonas vaginalis infection, the kinetics of antibody levels decrease following therapy has never been studied. We thus investigated the clearance of circulating anti-T. vaginalis IgGs after pharmacological treatment in patients affected by trichomoniasis. METHODS: 18 female patients affected by acute trichomoniasis were enrolled in this study. After metronidazole therapy administration, subjects were followed up monthly up to 5 months, and serum levels of anti-T. vaginalis IgGs were measured by ELISA. RESULTS: We showed that a successful therapy is characterised by a relatively fast decline of specific antibodies, until turning into negative by ELISA in 1-3 months. In a few patients we observed that the persistence of anti-T. vaginalis antibodies was associated with an evolution to chronic infection, which may be due to treatment failure or to reinfection by untreated sexual partners. CONCLUSIONS: Our results describe the direct correlation between the decline of a specific humoral anti-T. vaginalis response and an effective antimicrobial therapy. These findings may facilitate the follow-up approach to circumvent limitations in developing new diagnostic tools and techniques routinely used in microbiology laboratories to assess the presence of T. vaginalis in clinical samples.


Assuntos
Anticorpos Antiprotozoários/sangue , Antiprotozoários/uso terapêutico , Metronidazol/uso terapêutico , Tricomoníase/diagnóstico , Trichomonas vaginalis/imunologia , Vagina/parasitologia , Adulto , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina G/sangue , Cinética , Resultado do Tratamento , Tricomoníase/tratamento farmacológico , Tricomoníase/imunologia , Trichomonas vaginalis/patogenicidade
6.
Diagn Microbiol Infect Dis ; 110(2): 116479, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39116653

RESUMO

We report the use of a new multiplex Real-Time PCR platform to simultaneously identify 24 pathogens and 3 antimicrobial-resistance genes directly from respiratory samples of COVID-19 patients. Results were compared to culture-based diagnosis. Secondary infections were detected in 60% of COVID-19 patients by molecular analysis and 73% by microbiological assays, with no significant differences in accuracy, indicating Gram-negative bacteria as the predominant species. Among fungal superinfections, Aspergillus spp. were detected by both methods in more than 7% of COVID-19 patients. Oxacillin-resistant S. aureus and carbapenem-resistant K. pneumoniae were highlighted by both methods. Secondary microbial infections in SARS-CoV-2 patients are associated with poor outcomes and an increased risk of death. Since PCR-based tests significantly reduce the turnaround time to 4 hours and 30 minutes (compared to 48 hours for microbial culture), we strongly support the routine use of molecular techniques, in conjunction with microbiological analysis, to identify co/secondary infections.


Assuntos
COVID-19 , Coinfecção , SARS-CoV-2 , Humanos , COVID-19/diagnóstico , COVID-19/microbiologia , SARS-CoV-2/genética , Coinfecção/diagnóstico , Coinfecção/microbiologia , Coinfecção/virologia , Masculino , Pessoa de Meia-Idade , Feminino , Técnicas de Diagnóstico Molecular/métodos , Idoso , Infecções Respiratórias/microbiologia , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Multiplex/métodos , Adulto , Farmacorresistência Bacteriana/genética , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Bactérias/genética , Bactérias/efeitos dos fármacos , Bactérias/isolamento & purificação , Bactérias/classificação , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/microbiologia
7.
Sex Transm Infect ; 89(6): 449-54, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23633668

RESUMO

OBJECTIVES: Trichomonas vaginalis is the causative agent of trichomoniasis, one of the most common sexually transmitted diseases worldwide. In recent years we have described the symbiotic relationship between T vaginalis and Mycoplasma hominis. How this biological association might affect the pathogenicity of one or both the microorganisms is still unknown. Since local inflammation is thought to play a central role in T vaginalis infection, we investigated the in vitro response of human macrophages to naturally mycoplasma-free T vaginalis, as compared to a mycoplasma-infected trichomonad isolate. METHODS: THP-1 cells were stimulated with two isogenic T vaginalis isolates, one naturally mycoplasma-free and one stably associated with M hominis, and secreted cytokines measured by ELISA. Nuclear factor κB (NFκB) involvement in THP-1 response to T vaginalis and M hominis was evaluated by means of a reporter system based on detection of alkaline phosphatase activity. RESULTS: We found that the presence of M hominis upregulates the expression of a panel of proinflammatory cytokines in a synergistic fashion. We also found that the upregulation of the proinflammatory response by THP-1 cells involves the transcription factor NFκB. CONCLUSIONS: These findings suggest that the presence of M hominis in T vaginalis isolates might play a key role in inflammation during trichomoniasis, thus affecting the severity of the disease. The synergistic upregulation of the macrophage proinflammatory response might also affect some important clinical conditions associated with T vaginalis infection, such as the increased risk of acquiring cervical cancer or HIV, which are thought to be affected by the inflammatory milieu during trichomoniasis.


Assuntos
Citocinas/metabolismo , Inflamação/imunologia , Inflamação/patologia , Monócitos/imunologia , Mycoplasma hominis/imunologia , Trichomonas vaginalis/imunologia , Trichomonas vaginalis/microbiologia , Fosfatase Alcalina/análise , Linhagem Celular , Técnicas de Cocultura , Meios de Cultura/química , Ensaio de Imunoadsorção Enzimática , Genes Reporter , Humanos , Monócitos/microbiologia , Monócitos/parasitologia , Mycoplasma hominis/patogenicidade , Mycoplasma hominis/fisiologia , NF-kappa B/metabolismo , Simbiose , Trichomonas vaginalis/patogenicidade , Trichomonas vaginalis/fisiologia
8.
mBio ; 13(3): e0091822, 2022 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-35608298

RESUMO

Trichomonas vaginalis can host the endosymbiont Mycoplasma hominis, an opportunistic pathogenic bacterium capable of modulating T. vaginalis pathobiology. Recently, a new noncultivable mycoplasma, "Candidatus Mycoplasma girerdii," has been shown to be closely associated with women affected by trichomoniasis, suggesting a biological association. Although several features of "Ca. M. girerdii" have been investigated through genomic analysis, the nature of the potential T. vaginalis-"Ca. M. girerdii" consortium and its impact on the biology and pathogenesis of both microorganisms have not yet been explored. Here, we investigate the association between "Ca. M. girerdii" and T. vaginalis isolated from patients affected by trichomoniasis, demonstrating their intracellular localization. By using an in vitro model system based on single- and double-Mycoplasma infection of Mycoplasma-free isogenic T. vaginalis, we investigated the ability of the protist to establish a relationship with the bacteria and impact T. vaginalis growth. Our data indicate likely competition between M. hominis and "Ca. M. girerdii" while infecting trichomonad cells. Comparative dual-transcriptomics data showed major shifts in parasite gene expression in response to the presence of Mycoplasma, including genes associated with energy metabolism and pathogenesis. Consistent with the transcriptomics data, both parasite-mediated hemolysis and binding to host epithelial cells were significantly upregulated in the presence of either Mycoplasma species. Taken together, these results support a model in which this microbial association could modulate the virulence of T. vaginalis. IMPORTANCE T. vaginalis and M. hominis form a unique case of endosymbiosis that modulates the parasite's pathobiology. Recently, a new nonculturable mycoplasma species ("Candidatus Mycoplasma girerdii") has been described as closely associated with the protozoon. Here, we report the characterization of this endosymbiotic relationship. Clinical isolates of the parasite demonstrate that mycoplasmas are common among trichomoniasis patients. The relationships are studied by devising an in vitro system of single and/or double infections in isogenic protozoan recipients. Comparative growth experiments and transcriptomics data demonstrate that the composition of different microbial consortia influences the growth of the parasite and significantly modulates its transcriptomic profile, including metabolic enzymes and virulence genes such as adhesins and pore-forming proteins. The data on modulation from RNA sequencing (RNA-Seq) correlated closely with those of the cytopathic effect and adhesion to human target cells. We propose the hypothesis that the presence and the quantitative ratios of endosymbionts may contribute to modulating protozoan virulence. Our data highlight the importance of considering pathogenic entities as microbial ecosystems, reinforcing the importance of the development of integrated diagnostic and therapeutic strategies.


Assuntos
Mycoplasma , Tricomoníase , Trichomonas vaginalis , Ecossistema , Feminino , Humanos , Mycoplasma/genética , Mycoplasma hominis/genética , Tricomoníase/microbiologia , Trichomonas vaginalis/genética
9.
Antibiotics (Basel) ; 11(6)2022 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-35740218

RESUMO

Trichomoniasis, the most common non-viral sexually transmitted infection worldwide, is caused by the protozoon Trichomonas vaginalis. The 5- nitroimidazole drugs, of which metronidazole is the most prescribed, are the only effective drugs to treat trichomoniasis. Resistance against metronidazole is increasingly reported among T. vaginalis isolates. T. vaginalis can establish an endosymbiosis with two Mycoplasma species, Mycoplasma hominis and Candidatus Mycoplasma girerdii, whose presence has been demonstrated to influence several aspects of the protozoan pathobiology. The role of M. hominis in T. vaginalis resistance to metronidazole is controversial, while the influence of Ca. M. girerdii has never been investigated. In this work, we investigate the possible correlation between the presence of Ca. M. girerdii and/or M. hominis and the in vitro drug susceptibility in a large group of T. vaginalis isolated in Italy and in Vietnam. We also evaluated, via RNA-seq analysis, the expression of protozoan genes involved in metronidazole resistance in a set of syngenic T. vaginalis strains, differing only for the presence/absence of the two Mycoplasmas. Our results show that the presence of M. hominis significantly increases the sensitivity to metronidazole in T. vaginalis and affects gene expression. On the contrary, the symbiosis with Candidatus Mycoplasma girerdii seems to have no effect on metronidazole resistance in T. vaginalis.

10.
Microbiology (Reading) ; 156(Pt 12): 3734-3743, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20656780

RESUMO

Both Mycoplasma hominis and Trichomonas vaginalis utilize arginine as an energy source via the arginine dihydrolase (ADH) pathway. It has been previously demonstrated that M. hominis forms a stable intracellular relationship with T. vaginalis; hence, in this study we examined the interaction of two localized ADH pathways by comparing T. vaginalis strain SS22 with the laboratory-generated T. vaginalis strain SS22-MOZ2 infected with M. hominis MOZ2. The presence of M. hominis resulted in an approximately 16-fold increase in intracellular ornithine and a threefold increase in putrescine, compared with control T. vaginalis cultures. No change in the activity of enzymes of the ADH pathway could be demonstrated in SS22-MOZ2 compared with the parent SS22, and the increased production of ornithine could be attributed to the presence of M. hominis. Using metabolic flow analysis it was determined that the elasticity of enzymes of the ADH pathway in SS22-MOZ2 was unchanged compared with the parent SS22; however, the elasticity of ornithine decarboxylase (ODC) in SS22 was small, and it was doubled in SS22-MOZ2 cells. The potential benefit of this relationship to both T. vaginalis and M. hominis is discussed.


Assuntos
Arginina/metabolismo , Mycoplasma hominis/metabolismo , Trichomonas vaginalis/metabolismo , Trichomonas vaginalis/microbiologia , Sequência de Aminoácidos , Hidrolases/química , Hidrolases/genética , Hidrolases/metabolismo , Dados de Sequência Molecular , Ornitina Descarboxilase/química , Ornitina Descarboxilase/genética , Ornitina Descarboxilase/metabolismo , Proteínas de Protozoários/química , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Alinhamento de Sequência , Trichomonas vaginalis/enzimologia , Trichomonas vaginalis/genética
11.
Front Microbiol ; 11: 616213, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33488560

RESUMO

Ever since the publication of the seminal paper by Lynn Margulis in 1967 proposing the theory of the endosymbiotic origin of organelles, the study of the symbiotic relationships between unicellular eukaryotes and prokaryotes has received ever-growing attention by microbiologists and evolutionists alike. While the evolutionary significance of the endosymbiotic associations within protists has emerged and is intensively studied, the impact of these relationships on human health has been seldom taken into account. Microbial endosymbioses involving human eukaryotic pathogens are not common, and the sexually transmitted obligate parasite Trichomonas vaginalis and the free-living opportunistic pathogen Acanthamoeba represent two unique cases in this regard, to date. The reasons of this peculiarity for T. vaginalis and Acanthamoeba may be due to their lifestyles, characterized by bacteria-rich environments. However, this characteristic does not fully explain the reason why no bacterial endosymbiont has yet been detected in unicellular eukaryotic human pathogens other than in T. vaginalis and Acanthamoeba, albeit sparse and poorly investigated examples of morphological identification of bacteria-like microorganisms associated with Giardia and Entamoeba were reported in the past. In this review article we will present the body of experimental evidences revealing the profound effects of these examples of protist/bacteria symbiosis on the pathogenesis of the microbial species involved, and ultimately their impact on human health.

12.
Front Cell Infect Microbiol ; 10: 581066, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33117734

RESUMO

Pore-forming proteins (PFPs) are a group of functionally versatile molecules distributed in all domains of life, and several microbial pathogens notably use members of this class of proteins as cytotoxic effectors. Among pathogenic protists, Entamoeba histolytica, and Naegleria fowleri display a range of pore-forming toxins belonging to the Saposin-Like Proteins (Saplip) family: Amoebapores and Naegleriapores. Following the genome sequencing of Trichomonas vaginalis, we identified a gene family of 12 predicted saposin-like proteins (TvSaplips): this work focuses on investigating the potential role of TvSaplips as cytopathogenetic effectors. We provide evidence that TvSaplip12 gene expression is potently upregulated upon T. vaginalis contact with target cells. We cloned and expressed recombinant TvSaplip12 in planta and we demonstrate haemolytic, cytotoxic, and bactericidal activities of rTvSaplip12 in vitro. Also, evidence for TvSaplip subcellular discrete distribution in cytoplasmic granules is presented. Altogether, our results highlight the importance of TvSaplip in T. vaginalis pathogenesis, depicting its involvement in the cytolytic and bactericidal activities during the infection process, leading to predation on host cells and resident vaginal microbiota for essential nutrients acquisition. This hence suggests a potential key role for TvSaplip12 in T. vaginalis pathogenesis as a candidate Trichopore.


Assuntos
Entamoeba histolytica , Trichomonas vaginalis , Entamoeba histolytica/genética , Feminino , Humanos , Porinas , Nicotiana , Trichomonas vaginalis/genética , Vagina
14.
J Pregnancy ; 2018: 5037181, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30174955

RESUMO

Mycoplasma hominis is considered an opportunistic pathogen able to colonize the lower urogenital tract; in females the infection is associated with severe pregnancy and postpartum complications, including abortion, endometritis, preterm delivery, and low birth weight. Molecular mechanisms of pathogenicity and virulence effectors remain poorly characterized. A number of studies in the last decade have demonstrated that M. hominis can establish an endosymbiotic relationship with Trichomonas vaginalis, a urogenital parasitic protozoon, also associated with adverse pregnancy outcomes. Recently, two bacterial genes (alr and goiB) associated with amniotic cavity invasion and a single gene (goiC) associated with intra-amniotic infections and high risk of preterm delivery have been identified in M. hominis isolated from a group of pregnant patients. In this work we demonstrate that a high number of M. hominis intracellularly associated with T. vaginalis have goiC gene, in association with alr and goiB. In addition, we demonstrate that metronidazole treatment of M. hominis-infected T. vaginalis allows delivering viable intracellular goiC positive M. hominis from antibiotic-killed protozoa and that free M. hominis can infect human cell cultures. Results suggest that molecular diagnostic strategies to identify both pathogens and their virulence genes should be adopted to prevent severe complications during pregnancy.


Assuntos
Antibacterianos/farmacologia , Metronidazol/farmacologia , Infecções por Mycoplasma/transmissão , Complicações Infecciosas na Gravidez/microbiologia , Trichomonas vaginalis/efeitos dos fármacos , Líquido Amniótico/microbiologia , Membranas Extraembrionárias/microbiologia , Feminino , Humanos , Mycoplasma hominis/isolamento & purificação , Trabalho de Parto Prematuro/microbiologia , Gravidez , Trichomonas vaginalis/imunologia , Trichomonas vaginalis/isolamento & purificação , Vagina/microbiologia
16.
Front Biosci ; 11: 2028-34, 2006 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-16720288

RESUMO

Mollicutes are the smallest and simplest self-replicating microorganisms. Despite the minimal genome and apparent lack of complexity, mycoplasmas show a high degree of adaptation to the most diverse environments. Mycoplasma hominis is a human sexually transmitted mycoplasma which is able to establish a biological association with Trichomonas vaginalis, a pathogenic flagellated protist. M. hominis and T. vaginalis share the same specific natural niche, the human genitourinary tract. Symbiotic relationships between unicellular eukaryotes and bacteria are well known and have been extensively studied, providing interesting insights into the biology of one or both the symbionts. The relationship between T. vaginalis and M. hominis is unique in that it was the first described association of two obligated human parasites. Several aspects of this relationship have been investigated, showing how the trichomonad may be viewed not only as a new niche for M. hominis, but also as a "Trojan horse" for the transmission of the bacterial infection to the human host.


Assuntos
Mycoplasma hominis/fisiologia , Trichomonas vaginalis/microbiologia , Animais , Feminino , Humanos , Infecções por Mycoplasma/complicações , Infecções por Mycoplasma/transmissão , Mycoplasma hominis/patogenicidade , Simbiose , Vaginite por Trichomonas/complicações , Vaginite por Trichomonas/microbiologia , Vaginite por Trichomonas/transmissão , Trichomonas vaginalis/fisiologia
17.
Front Microbiol ; 7: 953, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27379081

RESUMO

The symbiosis between the parasitic protist Trichomonas vaginalis and the opportunistic bacterium Mycoplasma hominis is the only one currently described involving two obligate human mucosal symbionts with pathogenic capabilities that can cause independent diseases in the same anatomical site: the lower urogenital tract. Although several aspects of this intriguing microbial partnership have been investigated, many questions on the influence of this symbiosis on the parasite pathobiology still remain unanswered. Here, we examined with in vitro cultures how M. hominis could influence the pathobiology of T. vaginalis by investigating the influence of M. hominis on parasite replication rate, haemolytic activity and ATP production. By comparing isogenic mycoplasma-free T. vaginalis and parasites stably associated with M. hominis we could demonstrate that the latter show a higher replication rate, increased haemolytic activity and are able to produce larger amounts of ATP. In addition, we demonstrated in a T. vaginalis-macrophage co-culture system that M. hominis could modulate an aspect of the innate immuno-response to T. vaginalis infections by influencing the production of nitric oxide (NO) by human macrophages, with the parasite-bacteria symbiosis outcompeting the human cells for the key substrate arginine. These results support a model in which the symbiosis between T. vaginalis and M. hominis influences host-microbes interactions to the benefit of both microbial partners during infections and to the detriment of their host.

18.
J Infect Dev Ctries ; 9(6): 655-60, 2015 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-26142677

RESUMO

INTRODUCTION: The diffusion of trichomoniasis in Vietnam has been scarcely studied. The aim of this study was to investigate the prevalence of trichomoniasis in a group of symptomatic and asymptomatic women in Central Vietnam. Relationships between education, socioeconomical and marital status, and sexual behavior with infection have also been investigated. METHODOLOGY: 249 symptomatic and 534 asymptomatic women from Hue City, Vietnam, were enrolled in this study. All women were interviewed about socioeconomical and behavioral status. They underwent clinical examination, and vaginal swabs were taken to assess T. vaginalis infection by wet mount microscopy examination. In addition, an ELISA test to detect antibodies to T. vaginalis in patients' sera was used. RESULTS: The overall prevalence of trichomoniasis assessed by microscopic examination was 6.6%. A significant difference between symptomatic and asymptomatic groups was observed, resulting in 19.3% and 0.7%, respectively. Anti- T. vaginalis antibodies were detected in 31.3% of symptomatic and in 13.3% of asymptomatic women. High-risk sexual behaviour, residence in urban areas, and low level of education were positively associated with infection. CONCLUSION: This is the first report on the diffusion of trichomoniasis in Central Vietnam on symptomatic and asymptomatic subjects. Data demonstrated that T. vaginalis is a common cause of vaginal infection in the Hue province. The prevalence detected by microscopic examination was high in symptomatic subjects, while serological ELISA test detected infection also in asymptomatic patients, who tested negative by microscopy. The ELISA test may be useful to detect infection, especially in asymptomatic population.


Assuntos
Anticorpos Antiprotozoários/sangue , Portador Sadio/epidemiologia , Vaginite por Trichomonas/epidemiologia , Trichomonas vaginalis/imunologia , Trichomonas vaginalis/isolamento & purificação , Adulto , Portador Sadio/parasitologia , Estudos Transversais , Escolaridade , Feminino , Humanos , Microscopia , Pessoa de Meia-Idade , Fatores de Risco , Estudos Soroepidemiológicos , Comportamento Sexual , Fatores Socioeconômicos , Vaginite por Trichomonas/parasitologia , Vietnã/epidemiologia , Adulto Jovem
19.
Vector Borne Zoonotic Dis ; 14(10): 716-23, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25325315

RESUMO

The recent characterization of the 18S ribosomal RNA (rRNA) of a pathogenic Babesia species in a domestic sow paved the way for establishing diagnostic and epidemiological tools for porcine babesiosis. Here, we developed the first specific Babesia sp. Suis PCR, and we applied this test to a panel of samples collected from animals living in a typical Mediterranean environment (Sardinia, Italy), including domestic pigs, wild boars, and ticks. In domestic pigs, PCR coupled with sequencing revealed an estimated Babesia infection frequency of 26.2% and the presence of distinct 18S sequence types. The different distribution of sequence types in symptomatic and asymptomatic subjects might suggest the existence of phylogenetically closely related strains with variable pathogenicity in pigs. Moreover, molecular identification of tick species indicated Rhipicephalus sanguineus and Rhipicephalus bursa as candidate vectors potentially involved in the transmission of this pathogen. Collectively, the data reveal the suitability of 18S rRNA PCR/sequencing for molecular diagnosis of porcine babesiosis and for large-scale investigations on the presence and geographical distribution of Babesia sp. Suis genetic variants.


Assuntos
Vetores Aracnídeos/parasitologia , Babesia/isolamento & purificação , Babesiose/diagnóstico , Doenças do Cão/diagnóstico , Rhipicephalus sanguineus/parasitologia , Doenças dos Suínos/diagnóstico , Animais , Babesia/classificação , Babesia/genética , Babesiose/parasitologia , Babesiose/transmissão , Sequência de Bases , DNA de Protozoário/química , DNA de Protozoário/genética , Doenças do Cão/parasitologia , Doenças do Cão/transmissão , Cães , Feminino , Itália/epidemiologia , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária , RNA Ribossômico 18S/genética , Sensibilidade e Especificidade , Análise de Sequência de DNA/veterinária , Especificidade da Espécie , Suínos , Doenças dos Suínos/parasitologia , Doenças dos Suínos/transmissão , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologia , Infestações por Carrapato/veterinária
20.
J Immunol Methods ; 382(1-2): 216-9, 2012 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-22683542

RESUMO

Synthetic peptides are widely used in indirect ELISA to detect and characterize specific antibodies in biological samples. Small peptides are not efficiently immobilized on plastic surfaces by simple adsorption, and the conjugation to carrier proteins with different binding techniques is the method of choice. Common techniques to conjugate peptide antigens to carrier proteins and to subsequently purify such complexes are time consuming, expensive, and occasionally abrogate immunogenicity of peptides. In this report we describe a simple, fast and inexpensive alternative protocol to immobilize synthetic peptides to plastic surfaces for standard ELISA. The technique is based on use of maleimide-activated bovine serum albumin or keyhole limpet hemocyanin as a protein anchor adsorbed on the polystyrene surface of the microtiter plate. Following adsorption of the carrier protein, sulfhydryl-containing peptides are cross-linked with an in-well reaction, allowing their correct orientation and availability to antibody binding, avoiding the time consuming steps needed to purify the hapten-carrier complexes. The immunoreactivity of peptides was tested by using both monoclonal and polyclonal antibodies in standard ELISA assays, and compared with established coating methods.


Assuntos
Ensaio de Imunoadsorção Enzimática/economia , Ensaio de Imunoadsorção Enzimática/métodos , Peptídeos/química , Peptídeos/imunologia , Poliestirenos/química , Anticorpos/imunologia , Proteínas de Transporte/química , Cisteína/química , Maleimidas/química , Propriedades de Superfície
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