ompA Sequencing and Multilocus Sequence Typing of Lymphogranuloma Venereum Cases in Buenos Aires Reveal New Chlamydia trachomatis Genotypes.

Since the Lymphogranuloma venereum (LGV) outbreak was first described in Buenos Aires in 2017, the detected strains presented peculiar characteristics. Our goal was to increase the understanding of the strains involved in the LGV outbreak in Argentina. We characterized the ompA gene sequences, using Sanger sequencing, of 88 LGV strains from 239 symptomatic patients in Buenos Aires enrolled between 2017 and 2019, and selected 20 C. trachomatis strains for further characterization using Multilocus Sequence Typing (MLST). Following the ompA gene analysis of the 88 LGV strains, we detected 43% L2b, 31% L1-like, and 26% L2. Among the 38 L2b samples analyzed, there were 7 distinct sequences, 3 of them not previously reported (L2bv12, L2bv13, and L2bv14). Additionally, we detected a strain with a new mutation (AM884176.1:g.59122A>T) found in the position defining L2 or L2b, proposed as L2i. Using MLST, five different sequence types (STs) were detected, including the ST2 (corresponding to the L1-like strains) and a new one (ST60). ST58 was associated with the concomitant presence of another STI and HIV. A high genetic diversity in C. trachomatis LGV strains in Argentina was observed in a short period of time, with a relatively low number of samples from a limited geographical area.

Impact of a public health intervention for active surveillance and mitigation of SARS-CoV-2 in a district from Buenos Aires province, Argentina: a descriptive epidemiological study.

BACKGROUND: The first case of SARS-CoV-2 was reported in Argentina on 3 March 2020. Measures to limit the spread of the virus were implemented, including complete lockdown (26 March). Nonetheless, the virus spread throughout the country, with a first peak of almost a million cases in October. On 30 November, the government's recommendation switched from social, preventive and compulsory isolation, to social, preventive and compulsory distancing. OBJECTIVES: To describe a tailored public health strategy to mitigate the spread of SARS-CoV-2 and determine its behaviour in San Antonio de Areco district from Buenos Aires province (Argentina) through a private-public association. DESIGN, SETTING AND PARTICIPANTS: Surveillance of the virus was performed with the local healthcare system, through early identification of cases and the systematic study of each infected individual and contact, regardless of symptomatology, using telemedicine and a COVID-19-specific outpatient clinic. Real-time PCR was used for detection using both individual and pooled samples, with a 12-hour turnaround time. RESULTS: Up to 30 November, a total of 2426 suspected cases were analysed and 578 were confirmed. Surveillance of health personnel and at-risk populations proved effective, mitigating viral spread. Pooling samples allowed reduction of operator time, helped reduce costs, and allowed detection of both symptomatic and asymptomatic cases. CONCLUSION: After 8 months of protocol implementation, the strategy to intensively survey groups at higher epidemiological risk and the systematic search for asymptomatic cases with the incorporation of pooled PCR for diagnosis, in combination with individual testing, is an efficient and viable option in populations with similar characteristics, in the frame of social isolation.

In vitro inhibitory effect of Hydrocotyle bonariensis Lam. extracts over Chlamydia trachomatis and Chlamydia pneumoniae on different stages of the chlamydial life cycle.

Chlamydial infections in humans are widely distributed and are responsible for a variety of acute and chronic diseases. Both Chlamydia trachomatis and Chlamydia pneumoniae can lead to chronic conditions that have been linked to complications and sequelae. This study aimed to develop a culture method in order to detect in vitro antichlamydial activity of different extracts obtained from native Argentinian plants used as antimicrobials in local ethnomedicine and to evaluate their inhibitory activity over Chlamydia trachomatis and Chlamydia pneumoniae growth. The inhibitory activity over different stages of the chlamydial life cycle on cell culture was assessed: the entry, the inclusion developing after entry, and the exponential growth stage. Also, the capability of rendering the cell refractory to chlamydial infection by pre-incubation with the extracts was assayed. Inhibitory activity of water-based and organic-based extracts obtained from Hydrocotyle bonariensis Lam. (Araliaceae), Lithraea molleoides (Vell.) Engl. (Anacardiaceae) and Hybanthus parviflorus (Mutis ex L.f.) Baill. (Violaceae) were tested against five strains of Chlamydia trachomatis (L2/434/BU and four clinical isolates form both neonatal conjunctivitis and adult genital infections, genotypes D, E, and K) and against Chlamydia pneumoniae AR39. The Hydrocotyle bonariensis dichloromethane extract showed a broad inhibitory activity over the exponential growth stage of Chlamydia trachomatis and Chlamydia pneumoniae independently from the chlamydial strain and the cell line. These results suggest a high inhibitory potential on both Chlamydiae species. In order to characterize the Hydrocotyle bonariensis dichloromethane active extract, an 1H-NMR was performed. The 1H-NMR characterization showed a spectrum with characteristic signals of the fatty acid moiety of lipids or cerebrosides, volatile phenolics, phytosterols, methyl triterpenes signals, and glucose moiety of the cerebrosides.

Proctitis infecciosa por Chlamydia Trachomatis / Infectious proctitis by Chlamydia Trachomatis

Introducción: Las infecciones transmisibles sexualmente (ITS) son motivo de consulta frecuente, encontrándose Chlamydia trachomatis (CT) entre las prevalentes. Este germen provoca proctitis de diversa gravedad según el biovar involucrado. Los casos más floridos suelen ser ocasionados por el biovar LGV, responsable de la entidad linfogranuloma venéreo. Se desconocen la prevalencia de CT como causa de proctitis en Argentina y los biovares implicados. Con el objetivo de estudiar estas variables, se diseñó un protocolo para detectar y genotipificar CT en pacientes con proctitis infecciosa. Pacientes y métodos: Se incluyeron pacientes mayores de 18 años con cuadro de proctitis infecciosa atendidos en un centro público y otro privado. Se excluyeron pacientes con enfermedad inflamatoria intestinal y radioterapia pelviana. El estudio fue aprobado por un Comité de Ética y los pacientes firmaron un consentimiento informado. En las muestras de hisopado anal se realizó detección y tipificación molecular de CT. Resultados: Entre 31de agosto de 2017 y 31 de mayo de 2018, se incluyeron 56 pacientes (1 mujer, 53 hombres, 2 mujeres trans), 79% HIV+. En 29 casos (52%) se detectó CT. Todos eran hombres que tienen sexo con hombres (HSH) y refirieron practicar sexo anal u oral receptivo no protegido. La mediana de edad de este subgrupo fue de 31 años; 83% HIV+ en tratamiento antirretroviral y mediana de CD4 637 cel/mm3. La coinfección con otras ITS fue del 41% (siendo las más frecuentes HPV, gonococia y sífilis). Los motivos de consulta más frecuentes fueron proctorragia, pujo y tenesmo, proctalgia y secreción. Las manifestaciones clínicas fueron variadas: proctitis, úlcera perianal, tumor endoanal/rectal y absceso/fístula. El 86% de las proctitis correspondió al biovar LGV, siendo 62% moderadas a graves. La mediana de tiempo de evolución hasta el diagnóstico fue 21 días. Los casos más prolongados correspondieron a cuadros clínicos y endoscópicos más graves. La duración del tratamiento se adecuó al biovar involucrado. Todos los pacientes respondieron favorablemente; sin embargo, las dos fístulas perianales requirieron resolución quirúrgica. Conclusiones: Proctitis, úlceras y fístulas son manifestaciones inespecíficas; el hallazgo clínico y endoscópico per se no son suficientes para definir la etiología; sólo una anamnesis minuciosa permite presumir una ITS como agente causal. La tipificación logra definir el biovar, dato fundamental para adecuar el tratamiento, cortar la cadena de transmisión y contar con datos epidemiológicos a nivel local. Como resultado de esta investigación, el Ministerio de Salud de Nación proyectó la emisión de una alerta sobre la presencia de LGV en nuestro medio. Tipo de estudio: Observacional, transversal, analítico, multicéntrico.

Introduction: Sexually transmitted infections (STI) are a frequent reason for consultation, being Chlamydia trachomatis (CT) among the most prevalent ones. It causes proctitis of varying severity depending on the biovar involved. The most severe cases are usually caused by the LGV biovar, responsible for the entity called lymphogranuloma venereum. The prevalence of CT as a cause of proctitis in Argentina and the biovars involved are unknown. In order to study these variables, a protocol was designed to detect and genotype CT in patients with infectious proctitis. Patients and methods: Patients over 18 years old with infectious proctitis were attended in a public and private center. Patients with inflammatory bowel disease and pelvic radiation therapy were excluded. The study was approved by an Ethics Committee and the patients signed an informed consent. The detection and molecular typing of CT was performed in anal swab samples. Results: Between 31-08-2017 and 31-05-2018, 56 patients were included (1 woman, 53 men, 2 trans women), 79% HIV +. In 29 cases (52%) CT was detected. All were MSM and reported to practice unprotected receptive oral or anal sex. The median age of this subgroup was 31 years; 83% HIV + on antiretroviral treatment and median CD4 637 cel / mm3. The coinfection with other STIs was present 41% (the most frequent were HPV, gonococcal and syphilis). The most frequent symptoms were bleeding, tenesmus, proctalgia and secretion. The clinical manifestations were varied: proctitis, perianal ulcer, endoanal / rectal tumor and abscess / anal fistula. 86% of the proctitis corresponded to the LGV biovar, being 62% moderate to severe. The median time of evolution until the diagnosis was 21 days. The most prolonged cases corresponded to more severe clinical and endoscopic symptoms. The duration of the treatment was adapted to the biovar involved. All patients responded favorably; however, the two perianal fistulas required surgical resolution. Conclusions: Proctitis, ulcers and fistulas are nonspecific manifestations; the clinical and endoscopic findings per se are not sufficient to define the etiology; only a meticulous anamnesis allows us to presume an STI as a causative agent. The typification allows to define the biovar, a fundamental data to adapt the treatment, stop chain of transmission and provides local epidemiological data. As a result of this investigation, the Ministry of Health of the Argentina issued an alert about the presence of LGV in our country. Type of study: Observational, cross-sectional, analytical, multicenter study.

Rectal Lymphogranuloma Venereum, Buenos Aires, Argentina.

Among 34 men with proctitis in Buenos Aires, Argentina, 16 (47%) had Chlamydia trachomatis infection, 11 (68.8%) of which were biovar lymphogranuloma venereum. The outbreak was probably local, as in Europe. In Argentina, lymphogranuloma venereum should be a suspected cause of proctitis in HIV-infected men who have had unprotected anal sex with men.

Optimization of Trichomonas vaginalis Diagnosis during Pregnancy at a University Hospital, Argentina.

The aim of this study was to evaluate different methods for Trichomonas vaginalis diagnosis during pregnancy in order to prevent maternal and perinatal complications. A total of 386 vaginal exudates from pregnant women were analyzed. T. vaginalis was investigated by 3 types of microscopic examinations direct wet mount with physiologic saline solution, prolonged May-Grunwald Giemsa (MGG) staining, and wet mount with sodium-acetate-formalin (SAF)/methylene blue method. PCR for 18S rRNA gene as well as culture in liquid medium were performed. The sensitivity and specificity of the microscopic examinations were evaluated considering the culture media positivity or the PCR techniques as gold standard. The frequency of T. vaginalis infection was 6.2% by culture and/or PCR, 5.2% by PCR, 4.7% by culture, 3.1% by SAF/methylene blue method and 2.8% by direct wet smear and prolonged MGG staining. The sensitivities were 83.3%, 75.0%, 50.0%, and 45.8% for PCR, culture, SAF/methylene blue method, and direct wet smear-prolonged MGG staining, respectively. The specificity was 100% for all the assessed methods. Microscopic examinations showed low sensitivity, mainly in asymptomatic pregnant patients. It is necessary to improve the detection of T. vaginalis using combined methods providing higher sensitivity, such as culture and PCR, mainly in asymptomatic pregnant patients, in order to prevent maternal and perinatal complications.

Prevalencia de Chlamydia trachomatis en la población que se atiende en un hospital universitario / Prevalence of Chlamydia trachomatis in a population that attend an university hospital

Chlamydia trachomatis es uml.MLn importante agente de transmisión sexual. La prevalencia varía de acuerdo a la población en estudio. El objetivo de éste trabajo es conocer la prevalencia de infección por C. trachomatis en los pacientes atendidos en el Hospital de Clínicas José de San Martín. Desde el 1§ de enero de 1999 al 20 de octubre de 2000 se analizaron retrospectivamente los especímenes de pacientes para investigación de C. trachomatis en el Laboratorio de Bacteriología Clínica, Depto. de Bioquímica Clínica del Hospital de Clínicas. La búsqueda de C. trachomatis en mujeres se realizó a partir de hisopado endocervical (332) y en hombres, en orina 1§ chorro (220) e hisopados uretrales (33). Para el diagnóstico, se utilizó inmunoensayo óptico Chlamyfast OIA (International, Microbio, Signes, Francia) en todas las muestras y en hisopados endocervicales y uretrales, además se realizaron cultivo en línea celular LLCMK2 y Nested PCR del gen omp-1. La edad promedio de las mujeres fue 33 años (15-72 años) y de los hombres 35 años (19-70 años). En el 50 por ciento de las mujeres el motivo de consulta fue infertilidad, mientras que en el hombre ésta consulta representó el 30 por ciento. La prevalencia de C. trachomatis fue 2,4 por ciento (6/253) en hombres y 0,9 por ciento (3/332) en mujeres. En las 3 muestras endocervicales se detectó C. trachomatis por Nested PCR, cultivo en 1 de 3 y no se detectó en ninguna de las muestras por inmunoensayo óptico. En las muestras provenientes de pacientes masculinos el diagnóstico se realizó por inmunoensayo óptico en todas las muestras y en hisopados uretrales: Nested PCR 2, cultivo celular 1. La prevalencia de C. trachomatis en la población estudiada es baja. Estos resultados permiten suponer una baja circulación de C. trachomatis en la población estudiada

Determinación de la concentración inhibitoria mínima de azitromicina para Chlamydia trachomatis en distintas líneas celulares / Determination of the minimun inhibitory concentration of azythromicin for chlamydia trachomatis in different cells cultures

Las bacterias del orden Chlamydiales son parásitos intracelulares obligados responsables de diferentes patologías tanto en humanos como en animales. Esta característica biológica hace que la determinación de la Concentración Inhibitoria Mínima (CIM) presente dificultades en su realización e interpretación. Se carece de una metodología normatizada, lo que dificulta el análisis y homologación de los pocos resultados publicados. Entre las múltiples variables a tener en cuenta, es importante considerar la línea celular a utilizar. En éste trabajo, se ha determinado la CIM de azitromicina para la cepa de Chlamydia trachomatis L2/434/BU utilizando las líneas celulares LLCMK2, McCoy y Vero, detectándose diferencias destacables en cuánto a los valores obtenidos. Se sugiere como el sustrato celular de mayor reproducibilidad a la línea LLCMK2 y se propone la normatización de los otros parámetros necesarios para determinar la CIM de las bacterias de éste orden