Epidemiological and molecular characteristics of Chlamydia psittaci from 8 human cases of psittacosis and 4 related birds in Argentina.

In Argentina, the epidemiological and molecular characteristics of Chlamydia psittaci infections are still not sufficiently known. A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for C. psittaci from January 2010 to March 2015. Four samples of birds related to these patients were also studied. Forty-eight samples were positive for C. psittaci by a nested PCR. The molecular characterization of twelve C. psittaci PCR-positive samples received in the National Reference Laboratory INEI-ANLIS "Dr. Carlos G. Malbrán", Buenos Aires, Argentina was performed. Eight positive samples from humans and four from birds were genotyped by ompA gene sequencing. C. psittaci genotype A was found in all human samples and in the related birds. This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of C. psittaci to conduct effective surveillance of its zoonotic infections.

Genetic diversity of Chlamydia among captive birds from central Argentina.

To study the occurrence of Chlamydia spp. and their genetic diversity, we analysed 793 cloacal swabs from 12 avian orders, including 76 genera, obtained from 80 species of asymptomatic wild and captive birds that were examined with conventional nested polymerase chain reaction and quantitative polymerase chain reaction. Chlamydia spp. were not detected in wild birds; however, four species (Chlamydia psittaci, Chlamydia pecorum, Chlamydia pneumoniae and Chlamydia gallinacea) were identified among captive birds (Passeriformes, n = 20; Psittaciformes, n = 15; Rheiformes, n = 8; Falconiformes n = 2; Piciformes n = 2; Anseriformes n = 1; Galliformes n = 1; Strigiformes n = 1). Two pathogens (C. pneumoniae and C. pecorum) were identified simultaneously in samples obtained from captive birds. Based on nucleotide-sequence variations of the ompA gene, three C. psittaci-positive samples detected were grouped into a cluster with the genotype WC derived from mammalian hosts. A single positive sample was phylogenetically related to a new strain of C. gallinacea. This report contributes to our increasing understanding of the abundance of Chlamydia in the animal kingdom.

Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina.

In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89%) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina.

Expression of HPV-16 E6 and E7 oncoproteins alters Chlamydia trachomatis developmental cycle and induces increased levels of immune regulatory molecules.

Introduction: Infection with Human Papillomavirus (HPV) is a recognized risk factor for Chlamydia trachomatis (CT) infection and vice versa. Coinfection of HPV and CT in women is a very common and usually asymptomatic finding that has been linked to increased risk of cervical cancer. It has been demonstrated that CT facilitates the entry of multiple high risk HPV genotypes, leading to damage of the mucosal barrier and interfering with immune responses and viral clearance, which ultimately favours viral persistence and malignant transformation. Although the facilitating effects elicited by CT infection on viral persistence have been reported, little is known about the consequences of HPV infection on CT development. Methods: Herein, we took advantage of a genetically modified human cervical cell line co-expressing HPV-16 major oncogenic proteins E6 and E7, as an experimental model allowing to investigate the possible effects that HPV infection would have on CT development. Results and discussion: Our results show that CT infection of HPV-16 E6E7 expressing cells induced an upregulation of the expression of E6E7 oncoproteins and host cell inhibitory molecules PD-L1, HVEM and CD160. Additionally, smaller chlamydial inclusions and reduced infectious progeny generation was observed in E6E7 cells. Ultrastructural analysis showed that expression of E6 and E7 did not alter total bacterial counts within inclusions but resulted in increased numbers of reticulate bodies (RB) and decreased production of infectious elementary bodies (EB). Our results indicate that during CT and HPV coinfection, E6 and E7 oncoproteins impair RB to EB transition and infectious progeny generation. On the other hand, higher expression of immune inhibitory molecules and HPV-16 E6E7 are cooperatively enhanced in CT-infected cells, which would favour both oncogenesis and immunosuppression. Our findings pose important implications for clinical management of patients with HPV and CT coinfection, suggesting that screening for the mutual infection could represent an opportunity to intervene and prevent severe reproductive health outcomes, such as cervical cancer and infertility.

Association between Human Papillomavirus and Chlamydia trachomatis genital infections in male partners of infertile couples.

The prevalence of HPV infection and its relationship with other sexually transmitted infections was analyzed in a cohort of 117 male partners of infertile couples from Cordoba, Argentina. Semen samples and urethral swabs were obtained and the infection with HPV, Chlamydia trachomatis, HSV1, HSV2, Mycoplasma hominis and Ureaplasma urealyticum was analyzed. A prevalence of HPV infection of 27.4% was found. Interestingly, infections by exclusively low risk HPV genotypes or high/intermediate risk HPV genotypes were present in 64.5% and 22.6% of cases, respectively. Low risk-HPV6 was the most frequently detected genotype. Remarkably, HPV and C. trachomatis infections were significantly associated to each other (OR: 11.55, 95% CI 1.14-117.06). No significant differences in sperm quality were found between HPV-positive and HPV-negative patients indicating that HPV male urogenital infection does not impair sperm quality. Our results show a high prevalence of HPV urogenital infection among male partners of infertile couples, and that HPV and C. trachomatis infections are reciprocal risk factors of their co-infection. Moreover, our results suggest that men constitute a reservoir for continued transmission of C. trachomatis and HPV to women highlighting the need for routine screening for these two pathogens in male partners of infertile couples.

Sexually transmitted infections in oral cavity lesions: Human papillomavirus, Chlamydia trachomatis, and Herpes simplex virus.

Objective: Provide evidence of HPV, C. trachomatis, and HSV infection in the oral cavity from patients with different types of stomatological lesions. Materials and Methods: Oral swabs samples were collected from a total of 318 patients. The infectious agents were analyzed using the PCR technique. HPV genotyping and HSV type were studied using the RFLP method. Results: We studied 137 benign lesions (B), 96 potentially malignant disorders (PMD) and 85 oral squamous cell carcinomas (OSCC). The prevalence of HPV was 34%. The most frequently genotypes detected were 6 low risk and 16 high risk. The prevalence of C. trachomatis was 16% and HSV 3%. Co-infections were detected mostly in benign lesions as following: HPV-C. trachomatis in 4%, C. trachomatis- HSV in 1.8% and HPV-HSV in 0.3%. Conclusion: This report is the first contribution to the identification and genotype characterization of HPV in a scenario little studied in our area, and it also contributes to improving our understanding on sexually transmitted infectious agents and their associations with the oral cavity. Besides, we detect the presence of C. trachomatis and HSV and co-infection with HPV in the oral cavity, which they should be taken into account for diagnostic and treatment purposes.

Molecular characterization of Chlamydia pneumoniae in animals and humans from Argentina: Genetic characterization of Chlamydia pneumoniae.

In this study, genetic diversity of Chlamydia pneumoniae was investigated and the relationships between sequences amplified of different sources, clinical conditions and geographical regions of central Argentina were established. Samples amplified were similar to human C. pneumoniae patterns and show the high clonality of the population.

Mutation detection of E6 and LCR genes from HPV 16 associated with carcinogenesis.

Human papillomavirus (HPV) is responsible for one of the most frequent sexually transmitted infections. The first phylogenetic analysis was based on a LCR region fragment. Nowadays, 4 variants are known: African (Af-1, Af-2), Asian-American (AA) and European (E). However the existence of sub-lineages of the European variant havs been proposed, specific mutations in the E6 and LCR sequences being possibly related to persistent viral infections. The aim of this study was a phylogenetic study of HPV16 sequences of endocervical samples from Cordoba, in order to detect the circulating lineages and analyze the presence of mutations that could be correlated with malignant disease. The phylogenetic analysis determined that 86% of the samples belonged to the E variant, 7% to AF-1 and the remaining 7% to AF-2. The most frequent mutation in LCR sequences was G7521A, in 80% of the analyzed samples; it affects the binding site of a transcription factor that could contribute to carcinogenesis. In the E6 sequences, the most common mutation was T350G (L83V), detected in 67% of the samples, associated with increased risk of persistent infection. The high detection rate of the European lineage correlated with patterns of human migration. This study emphasizes the importance of recognizing circulating lineages, as well as the detection of mutations associated with high-grade neoplastic lesions that could be correlated to the development of carcinogenic lesions.

Epidemiological and molecular characteristics of Chlamydia psittaci from 8 human cases of psittacosis and 4 related birds in Argentina / Características epidemiológicas y moleculares de Chlamydia psittaci provenientes de 8 casos humanos de psitacosis y de 4 aves relacionadas en la Argentina

In Argentina, the epidemiological and molecular characteristics of Chlamydia psittaci infections are still not sufficiently known. A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for C. psittaci from January 2010 to March 2015. Four samples of birds related to these patients were also studied. Forty-eight samples were positive for C. psittaci by a nested PCR. The molecular characterization of twelve C. psittaci PCR-positive samples received in the National Reference Laboratory INEI-ANLIS "Dr. Carlos G. Malbrán", Buenos Aires, Argentina was performed. Eight positive samples from humans and four from birds were genotyped by ompA gene sequencing. C. psittaci genotype A was found in all human samples and in the related birds. This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of C. psittaci to conduct effective surveillance of its zoonotic infections.

En la Argentina, aún no se conocen suficientemente las características epidemiológicas y moleculares de las infecciones por Chlamydia psittaci. Entre enero del 2010 y marzo del 2015 se estudiaron 846 muestras respiratorias y 10 oculares de pacientes con sospecha de psitacosis para la búsqueda de C. psittaci. También se estudiaron 4 muestras de aves relacionadas con estos pacientes. De ese total, 48 muestras fueron positivas para C. psittaci mediante una reacción en cadena de la polimerasa (PCR) anidada. Posteriormente, se realizó en el INEI-ANLIS «Dr. Carlos G. Malbrán¼ la caracterización molecular de 12 muestras positivas para C. psittaci, 8 de humanos y 4 de aves, que fueron genotipificadas por secuenciación del gen ompA. C. psittaci genotipo A se encontró en todas esas muestras. Este informe contribuye a mejorar nuestro conocimiento de las características epidemiológicas y moleculares de C. psittaci para lograr una vigilancia efectiva de la zoonosis que produce.

Evidencia molecular de la infección por Chlamydophila pneumoniae en reptiles de Argentina / Molecular evidence of Chlamydophila pneumoniae infection in reptiles in Argentina

En la región central de Argentina, las características epidemiológicas y moleculares de las infecciones por Chlamydophila pneumoniae en reptiles son desconocidas. Para detectar C. pneumoniae, se usó la reacción en cadena de la polimerasa anidada que amplifica el gen rpoB en muestras de hisopado cloacal de 19 reptiles. Once (57,89 %) reptiles resultaron positivos. La secuenciación y el análisis filogenético corroboraron la presencia de esta bacteria. No se detectó ADN de C. pneumoniae en la faringe ni IgM anti-C. pneumoniae en el suero de los cuidadores; sin embargo, ellos presentaron títulos muy elevados de IgG anti-C. pneumoniae. La detección de ADN de C. pneumoniae en los reptiles demostró la circulación de este agente en el centro recreativo donde se realizó este estudio, lo que podría explicar la exacerbada respuesta inmunitaria en los cuidadores; este hallazgo sugiere la presencia de un potencial ciclo zoonótico. Se reporta aquí por primera vez la detección de C. pneumoniae en reptiles en Argentina

In the central area of Argentina, the epidemiological and molecular characteristics of Chlamydophila pneumoniae infections in reptiles are still unknown. A nested polymerase chain reaction of the rpoB gene was used to detect C. pneumoniae in cloacal swab samples from 19 reptiles at a recreational area. Eleven (57.89%) reptiles were positive; the sequencing and phylogenetic analysis confirmed the presence of this bacterium. Neither C. pneumoniae DNA in the caregivers'pharynges nor IgM antibodies anti-C. pneumoniae in their serum samples were detected; however, caregivers presented very high titers of IgG anti-C. pneumoniae. The detection of C. pneumoniae DNA in reptiles demonstrated the circulation of this agent in the recreational area and could be responsible for the exacerbated immune response of the personnel handling the reptiles, which suggests a potential zoonotic cycle. This is the first report of the detection of C. pneumoniae in reptiles in Argentina

Aislamiento de Chlamydia Trachomatis y respuesta inmune en diferentes poblaciones / Isolation of Chlamydia trachomatis and immune response in different populations

Se estudió la presencia de IgG específica para C. trachomatis en diferentes grupos de pacientes adultos y la presencia de IgM en los neonatos y se relacionaron con el aislamiento de la bacteria en cultivos celulares. Se empleó un método de IFI sobre células McCoy, infectadas con el serotipo L(2)434/Bu de C. trachomatis. El método demostró ser específico, sensible y reproducible, utilizando las placas de plástico de 24 pozos para hacer crecer las células. Encontramos IgG específica para C. trachomatis en el 27 por ciento de las mujeres con síntomas clínicos, en el 40 por ciento de las que concurrieron al control ginecológico, en el 60 por ciento de las que consultaron por esterilidad y en el 10 por ciento de las embarazadas. Con un test de hipótesis para comparar proporciones se demostró que los valores e la presencia de IgG específicas es altamente significativa (p < 0,0001) para las estériles con respecto a las emabarazadas. De la relación del aislamiento de la bacteria con la presencia de IgG específica se formaron 4 grupos de pacientes, 7 de 10 tuvieron aislamiento e IFI positivo, 5 de 8, aislamiento positivo e IFI negativo; 25 de 28 (grupo de embarazadas), aislamiento negativo e IFI positivo y 63 de 76 dieron negativo en las 2 pruebas. El test de McNemar indica que la presencia de IgG tiene un alto grado de significación en las mujeres estériles con respecto a los otros grupos con una p < 0,001. En los neonatos se detectó C. trachomatis por aislamiento en un 20 por ciento, y presencia de IgM específica en un 10 por ciento. En los neonatos, habría un mejor diagnóstico combinando las 2 técnicas. Nuestros resultados sugieren que la detección de la IgG específica no es un criterio suficiente para el diagnóstico, debiendo acompañarse con el aislamiento de la bacteria. Sólo habría presencia de IgG con valores diagnósticos en mujeres estériles con antecedentes de C. trachomatis. Estos resultados señalan la importancia de la detección de C. trachomatis en jóvenes para evitar la infertilidad, y en embarazadas para prevenir la infección neonatal y la posibilidad de nacimientos prematuros y bajo peso al nacer de los niños.

Importancia de la infección por la bacteria chlamydophila pneumoniae en las enfermedades cardíacas de origen aterosclerótico / Importance of the infection by the bacterium chlamyophila pneumoniae in the diseases cardiac of aterosclerotico origin

Considerar la aterosclerosis como una enfermedad inflamatoria, ha renovado el interes en el rol que agentes infecciosos como la Chlamydophila pneumoniae (C pneumoniae) pueden jugar en el desarrollo del ateroma. El objeto de esta tesis fue investigar la asociación entre la C pneumoniae y la enfermedad cardiovascular aterosclerótica (ECA). Este estudio demostro que existían con más frecuencia anticuerpos tipo IgA contraC pnumoniae en pacientes con ECA (IgA OR=1,856), indicando que el predominio de infección persistente con C pneumoniae era más alto en pacientes con ECA porque la presencia de anticuerpos tipo IgA refleja una infcción activa o persistente, pero los anticuerpos tipo IgG reflejan sólo una infección anterior con C pneumoniae. La infección por C pneumoniae fue más frecuente en fumadores que en aquellas personas que nunca tuvieron el hábito de fumar, sugiriendo que el tabaquismo predispone al desarrollo de una infección crónica de C pneumoniae