Molecular Study of Pneumocystis jirovecii in Respiratory Samples of HIV Patients in Chile.

Pneumocystis is an opportunistic fungus that causes potentially fatal pneumonia (PCP) in immunocompromised patients. The objective of this study was to determine the prevalence of P. jirovecii in HIV patients through phenotypic and molecular study, to investigate the genetic polymorphisms of P. jirovecii at the mitochondrial gene mtLSU and at the nuclear dihydropteroate synthase gene (DHPS), and by analysis of molecular docking to study the effect of DHPS mutations on the enzymatic affinity for sulfamethoxazole. A PCP prevalence of 28.3% was detected, with mtLSU rRNA genotypes 3 (33.3%) and 2 (26.6%) being the most common. A prevalence of 6.7% (1/15) mutations in the DHPS gene was detected, specifically at codon 55 of the amino acid sequence of dihydropteroate synthase. Molecular docking analysis showed that the combination of mutations at 55 and 98 codons is required to significantly reduce the affinity of the enzyme for sulfamethoxazole. We observed a low rate of mutations in the DHPS gene, and molecular docking analysis showed that at least two mutations in the DHPS gene are required to significantly reduce the affinity of dihydropteroate synthase for sulfamethoxazole.

Protein Modelling and Molecular Docking Analysis of Fasciola hepatica ß-Tubulin's Interaction Sites, with Triclabendazole, Triclabendazole Sulphoxide and Triclabendazole Sulphone.

PURPOSE: Fasciola hepatica is a globally distributed trematode that causes significant economic losses. Triclabendazole is the primary pharmacological treatment for this parasite. However, the increasing resistance to triclabendazole limits its efficacy. Previous pharmacodynamics studies suggested that triclabendazole acts by interacting mainly with the ß monomer of tubulin. METHODS: We used a high-quality method to model the six isotypes of F. hepatica ß-tubulin in the absence of three-dimensional structures. Molecular dockings were conducted to evaluate the destabilization regions in the molecule against the ligands triclabendazole, triclabendazole sulphoxide and triclabendazole sulphone. RESULTS: The nucleotide binding site demonstrates higher affinity than the binding sites of colchicine, albendazole, the T7 loop and pßVII (p < 0.05). We suggest that the binding of the ligands to the polymerization site of ß-tubulin can lead a microtubule disruption. Furthermore, we found that triclabendazole sulphone exhibited significantly higher binding affinity than other ligands (p < 0.05) across all isotypes of ß-tubulin. CONCLUSIONS: Our investigation has yielded new insight on the mechanism of action of triclabendazole and its sulphometabolites on F. hepatica ß-tubulin through computational tools. These findings have significant implications for ongoing scientific research ongoing towards the discovery of novel therapeutics to treat F. hepatica infections.

High Presence of NETotic Cells and Neutrophil Extracellular Traps in Vaginal Discharges of Women with Vaginitis: An Exploratory Study.

Infectious vaginitis is a microbiological syndrome of great importance in public health that affects millions of women worldwide. However, no studies have explored the phenomenon of the production of the neutrophil extracellular traps (NETs) that are released into the female reproductive tract in these pathologies. This study aimed to determine the presence of NETosis in vaginal discharges of women with bacterial vaginosis, candidiasis, and trichomoniasis by characterizing NETs. Extracellular DNA with neutrophil elastase and citrullinated histones was identified to confirm the NET components (n = 10). The concentration, phenotypes of NETs, and number of NETotic cells were determined. The results showed an increase in NETotic cells in women with Candida albicans (CA) and Trichomonas vaginalis (TV) and an increase in NETs in TV-induced vaginitis. Samples of CA- and TV-infected women showed different NET phenotypes (diffNETs, sprNETs, and aggNETs); diffNETs were found in high concentrations in samples with CA and were increased in three types of NETs in TV infections. Samples with intermediate microbiota and bacterial vaginosis showed increased NETotic cells while the intermediate microbiota presented a higher concentration of NETs. Therefore, alterations in the microbiota and the presence of fungal and parasitic infections are important stimuli for the activation and induction of NETosis, and their cytotoxic effects could enhance tissue damage.

Penicillium digitatum, First Clinical Report in Chile: Fungal Co-Infection in COVID-19 Patient.

Penicillium digitatum is one of the most important phytopathogens. It causes deterioration and rotting of citrus fruits, generating significant economic losses worldwide. As a human pathogen, it is extremely rare. We present a case of pulmonary co-infection in a patient diagnosed with pneumonia due to SARS-CoV-2. A 20-year-old female patient, primigravid, 36 weeks of gestation, without comorbidities, and diagnosed with severe pneumonia due to the SARS-CoV-2, showed rapid lung deterioration for which their pregnancy was interrupted by surgery. The patient was hospitalized in the Intensive Care Unit (ICU), connected to mechanical ventilation and receiving corticosteroids and antibiotics. The diagnosis of pulmonary fungal infection was made through bronchoalveolar lavage (BAL) culture, and the species identification was performed by sequencing of ß-tubulin. Phylogenetic analysis with related species was performed for the confirmation of species identification. Antifungal susceptibility tests were performed for itraconazole (4 µg/mL), voriconazole (2 µg/mL), and amphotericin B (2 µg/mL). The patient was successfully treated with itraconazole. This is the second worldwide report of pulmonary infection by P. digitatum and the first in Chile. Although it is a fungus that rarely infects humans, it could represent an emerging opportunistic fungal pathogen, with associated risk factors that should be considered in the differential diagnosis of Penicillium species isolated from infections in humans.

Concordance Between Clinical and Laboratory Diagnosis of Abnormal Vaginal Discharge in Chilean Women.

OBJECTIVE: To determine the concordance between the clinical diagnosis of women with abnormal vaginal discharge (AVD) and laboratory results using molecular detection and observation of the vaginal microbiota. METHODS: Cross-sectional study conducted in 2018 in Temuco, Chile. A total of 25 midwives from 12 health centers participated. A total of 125 women > 18 years old, volunteers, were recruited. The sample of the posterior vaginal fornix was obtained by speculoscopy. Characteristics of the discharge and of the external and internal genitalia were observed. Gram staining was used to observe vaginal microbiota, blastoconidia and pseudohyphae, and polymerase chain reaction was used for the detection of Trichomonas vaginalis and Candida albicans. The Cohen kappa coefficient was used in the concordance analysis. RESULTS: Out of a total of 125 women with AVD, 85.6% consulted spontaneously and 14.4% were diagnosed clinically during a routine check-up. Absolute concordance was significant (p = 0.0012), with an agreement of 13.6%. The relative concordance was significant, but fair for bacterial vaginosis (Kappa = 0.21; p = 0.003) and candidiasis (Kappa = 0.22; p = 0.001), and slight for trichomoniasis (Kappa = 0.14; p = 0.009). The percentage of coincidence of the diagnoses (single or mixed) by laboratory and midwives was: bacterial vaginosis 63.2% (12/19), candidiasis 36.5% (27/74), and trichomoniasis 12.5% (4/32). There was 20% coinfection. A total of 36% of the clinical diagnoses of AVD had negative laboratory tests. CONCLUSION: The vulvovaginitis conditions candidiasis and trichomoniasis appear to be overdiagnosed, and bacterial vaginosis appears to be underdiagnosed by the clinical diagnosis when compared with the laboratory diagnosis. The low concordance obtained shows the importance of complementing the clinical diagnosis with a laboratory study of AVD, particularly in women with failed treatments and/or coinfections with unspecific and varying signs and symptoms.

OBJETIVO: Determinar a concordância entre o diagnóstico clínico de mulheres com corrimento vaginal anormal (AVD) e os resultados laboratoriais por meio da detecção molecular e observação da microbiota vaginal. MéTODOS: Estudo transversal realizado em 2018 em Temuco, Chile. Participaram 25 parteiras de 12 centros de saúde. Um total de 125 mulheres > 18 anos, voluntárias, foram recrutadas. A amostra do fórnice vaginal posterior foi obtida por especuloscopia. Foram observadas características da secreção e da genitália externa e interna. A coloração de Gram foi usada para observar a microbiota vaginal, blastoconídios e pseudo-hifas, e a reação em cadeia da polimerase foi usada para a detecção de Trichomonas vaginalis e Candida albicans. O coeficiente kappa de Cohen foi usado na análise de concordância. RESULTADOS: De um total de 125 mulheres com AVD, 85,6% consultaram espontaneamente e 14,4% foram diagnosticados clinicamente durante um check-up de rotina. A concordância absoluta foi significativa (p = 0,0012), com concordância de 13,6%. A concordância relativa foi significativa, mas razoável para vaginose bacteriana (Kappa = 0,21; p = 0,003) e candidíase (Kappa = 0,22; p = 0,001), e leve para tricomoníase (Kappa = 0,14; p = 0,009). O percentual de coincidência dos diagnósticos (solteiros ou mistos) por laboratório e parteiras foi: vaginose bacteriana 63,2% (12/19), candidíase 36,5% (27/74) e tricomoníase 12,5% (4/32). Houve 20% de coinfecção. Um total de 36% dos diagnósticos clínicos de AVD tiveram exames laboratoriais negativos. CONCLUSãO: As condições de vulvovaginite candidíase e tricomoníase parecem ser sobrediagnosticadas, e a vaginose bacteriana parece ser subdiagnosticada pelo diagnóstico clínico quando comparado com o diagnóstico laboratorial. A baixa concordância obtida mostra a importância de complementar o diagnóstico clínico com estudo laboratorial de AVD, principalmente em mulheres com falha de tratamento e / ou coinfecções com sinais e sintomas inespecíficos e variáveis.

[Factors associated with Chlamydia trachomatis in women with vaginitis attending in primary health care]. / Factores asociados a Chlamydia trachomatis en mujeres con vaginitis atendidas en atención primaria en salud.

BACKGROUND: Chlamydia trachomatis is recognized as the causative agent of one of the most common and curable sexually transmitted infections (STIs). In women, it can be a trigger for pelvic inflammatory disease, a risk factor for infertility or acquisition of other STIs. AIM: To determine CT in women with vaginitis and associate it with sociodemographic, sexual, clinical, and vaginal microbiota factors. METHODS: 121 female volunteers > 18 years of age, with a clinical diagnosis of vaginitis and abnormal vaginal discharge, attending in primary health care services (PHC) in Temuco-Chile were recluted. A vaginal discharge sample was taken, determining C. trachomatis by polymerase chain reaction and vaginal microbiota by Gram stain. Clinical, sociodemographic and sexual data were collected by survey. RESULTS: Age range was 18-61 years. The frequency of C. trachomatis was 14.9%. C. trachomatis was associated with a higher number of sexual partners in the last 5 years (p = 0.001), oral plus anal sex (p = 0.055) and sex toys (p = 0.027); there was a non-significant association with bacterial vaginosis 31.2% (p = 0.091) and initiation of sexual activity < 18 years 20.6% (p = 0.07). CONCLUSIONS: It would be beneficial to incorporate in the gynecological semiology of the women attended in PHC services, the factors found to favor C. trachomatis infection, as this could help the management of this STI.

Humanized Care from the Perception of Oncology Patients from Southern Chile.

OBJECTIVES: This work sought to determine the perception of behaviors of humanized nursing care and its relation with sociodemographic and clinical variables in patients hospitalized in a Hemato-Oncology Department. METHODS: Analytic cross-sectional study conducted with 51 patients hospitalized in the Hemato-Oncology Unit at Hospital Base Valdivia, Chile. A survey containing sociodemographic and clinical information was applied together with the questionnaire on Perception of Behaviors of Humanized Nursing Care 3rd version" -PBHNC 3v (32 items distributed in the categories: Qualities of nursing work, Openness to nurse-patient communication, and Willingness to care). RESULTS: Of the participants, 51% were women, with mean age of 46.5±16.6 years; 54.9% were diagnosed with Lymphoma and 78.4% were in the treatment induction stage. In 30 of the 32 items of the instrument, > 90% of the participants evaluated compliance with the behavior of caring at level of "always". By categories, it was observed that for "Willingness to care" there was significantly lower score among patients from 18 to 49 years of age (p=0.0455). For the category "Openness to nurse-patient communication" lower median score existed in patients with Myeloma (p=0.0043) and in patients in the Remission-Consolidation stage (p=0.0084). Days of hospitalization were associated significantly with the category "Willingness to care", being lower with 16 days and more (p=0.0242). CONCLUSIONS: High frequency was observed of humanized-care behaviors and small differences in their assessment that were associated with demographic factors like age, and clinical factors, like diagnosis, treatment stage, and days of hospitalization.

Factores asociados a Chlamydia trachomatis en mujeres con vaginitis atendidas en atención primaria en salud / Factors associated with Chlamydia trachomatis in women with vaginitis attending in primary health care

INTRODUCCIÓN: Chlamydia trachomatis es reconocida como el agente causal de una de las infecciones de transmisión sexual (ITS) más común y curable. En mujeres, puede ser gatillante de enfermedad inflamatoria pélvica, un factor de riesgo para infertilidad o adquisición de otras ITS. OBJETIVO: Determinar C. trachomatis en mujeres con diagnóstico clínico de vaginitis asociando factores sociodemográficos, sexuales, clínicos y microbiota vaginal. PACIENTES Y MÉTODOS: Participaron 121 mujeres voluntarias > 18 años, con diagnóstico clínico de vaginitis y descarga vaginal anormal, atendidas en servicios de atención primaria en salud (APS) en Temuco-Chile. Se tomó muestra de flujo vaginal, determinándose C. trachomatis por una reacción de polimerasa en cadena convencional y microbiota vaginal por tinción de Gram. Se colectaron datos clínicos, sociodemográficos y sexuales mediante encuesta. RESULTADOS: Rango de edad 18-61 años. La frecuencia de C. trachomatis fue 14,9%. Se asoció C. trachomatis a mayor número de parejas sexuales en los últimos 5 años (p = 0,001), relaciones sexuales oral más anal (p = 0,055) y juguetes sexuales (p = 0,027). Asociación no significativa hubo con vaginosis bacteriana 31,2% (p = 0,091) e inicio de la actividad sexual < 18 años 20,6% (p = 0,07). CONCLUSIONES: Sería beneficioso incorporar en la semiología ginecológica de las mujeres atendidas en servicios de APS, los factores encontrados como favorecedores de infección por C. trachomatis, pues ello podría ayudar al manejo de esta ITS.

BACKGROUND: Chlamydia trachomatis is recognized as the causative agent of one of the most common and curable sexually transmitted infections (STIs). In women, it can be a trigger for pelvic inflammatory disease, a risk factor for infertility or acquisition of other STIs. AIM: To determine CT in women with vaginitis and associate it with sociodemographic, sexual, clinical, and vaginal microbiota factors. Methods: 121 female volunteers > 18 years of age, with a clinical diagnosis of vaginitis and abnormal vaginal discharge, attending in primary health care services (PHC) in Temuco-Chile were recluted. A vaginal discharge sample was taken, determining C. trachomatis by polymerase chain reaction and vaginal microbiota by Gram stain. Clinical, sociodemographic and sexual data were collected by survey. RESULTS: Age range was 18-61 years. The frequency of C. trachomatis was 14.9%. C. trachomatis was associated with a higher number of sexual partners in the last 5 years (p = 0.001), oral plus anal sex (p = 0.055) and sex toys (p = 0.027); there was a non-significant association with bacterial vaginosis 31.2% (p = 0.091) and initiation of sexual activity < 18 years 20.6% (p = 0.07). CONCLUSIONS: It would be beneficial to incorporate in the gynecological semiology of the women attended in PHC services, the factors found to favor C. trachomatis infection, as this could help the management of this STI.

Humanized Care from the Perception of Oncology Patients from Southern Chile / Cuidado humanizado desde la percepción de pacientes oncológicos del sur de Chile / Cuidado humanizado a partir da percepção de pacientes com câncer no sul do Chile

Objective. This work sought to determine the perception of behaviors of humanized nursing care and its relation with sociodemographic and clinical variables in patients hospitalized in a Hemato-Oncology Department. Methods. Analytic cross-sectional study conducted with 51 patients hospitalized in the Hemato-Oncology Unit at Hospital Base Valdivia, Chile. A survey containing sociodemographic and clinical information was applied together with the questionnaire on Perception of Behaviors of Humanized Nursing Care 3rd version" -PBHNC 3v (32 items distributed in the categories: Qualities of nursing work, Openness to nurse-patient communication, and Willingness to care). Results. Of the participants, 51% were women, with mean age of 46.5±16.6 years; 54.9% were diagnosed with Lymphoma and 78.4% were in the treatment induction stage. In 30 of the 32 items of the instrument, > 90% of the participants evaluated compliance with the behavior of caring at level of "always". By categories, it was observed that for "Willingness to care" there was significantly lower score among patients from 18 to 49 years of age (p=0.0455). For the category "Openness to nurse-patient communication" lower median score existed in patients with Myeloma (p=0.0043) and in patients in the Remission-Consolidation stage (p=0.0084). Days of hospitalization were associated significantly with the category "Willingness to care", being lower with 16 days and more (p=0.0242). Conclusion. High frequency was observed of humanized-care behaviors and small differences in their assessment that were associated with demographic factors like age, and clinical factors, like diagnosis, treatment stage, and days of hospitalization.

Objetivo. Determinar la percepción de comportamientos de cuidados humanizados de enfermería y su relación con variables sociodemográficas y clínicas en pacientes hospitalizados en una Unidad de Hemato-Oncología. Métodos. Estudio transversal analítico realizado con la participación de 51 pacientes hospitalizados en la Unidad de Hemato-Oncología del Hospital Base Valdivia, Chile. Se aplicó un formulario que contenía información sociodemográfica, clínica y el instrumento Percepción de Comportamientos de Cuidados Humanizados de Enfermería 3ª versión" -PCHE 3v (32 ítems distribuidos en las categorías: Cualidades del hacer de enfermería, Apertura a la comunicación enfermera(o)-paciente y Disposición para la atención). Resultados. De los participantes, 51% fueron mujeres, con una edad media de 46.5±16.6 años. Un 54.9% presentó diagnóstico de Linfoma y 78.4% estaba en etapa de Inducción del tratamiento. En 30 de los 32 ítems del instrumento, sobre 90% de los participantes evaluaron el cumplimiento del comportamiento de cuidado en nivel "siempre". Por categorías se observó que para "Disposición para la atención" hubo significativa menor puntuación entre los pacientes de 18 a 49 años (p=0.0455). Para la categoría "Apertura a la comunicación enfermera(o)-paciente" existió menor puntaje mediano en pacientes con Mieloma (p=0.0043) y en los pacientes en etapa de Remisión-Consolidación (p=0.0084). Los días de hospitalización se asociaron significativamente con la categoría "Disposición para la atención", siendo menor con 16 días y más (p=0.0242). Conclusión. Hubo alta frecuencia de comportamientos de cuidados humanizados y pequeñas diferencias en su valoración que se asociaron con factores demográficos como la edad y a factores clínicos como el diagnóstico, la etapa de tratamiento y los días de hospitalización.

Objetivo. Verificar a percepção dos comportamentos de cuidado humanizado de enfermagem e sua relação com variáveis sociodemográficas e clínicas em pacientes internados em uma Unidade de Hemato-Oncologia. Métodos. Estudo transversal analítico realizado com a participação de 51 pacientes internados na Unidade de Hemato-Oncologia da Base Hospitalar de Valdivia, Chile. Foi aplicado um formulário que continha informações sociodemográficas e clínicas e o instrumento Percepção de Comportamentos da Assistência Humanizada de Enfermagem 3ª versão -PCHE 3v (32 itens distribuídos nas categorias: Qualidades do trabalho de enfermagem, Abertura à comunicação do enfermeiro (a) - Paciente e Disponibilidade para o cuidado). Resultados. Dos participantes, 51% eram mulheres, com média de idade de 46.5±16.6 anos. 54.9% apresentavam diagnóstico de Linfoma e 78.4% encontravam-se na fase de indução do tratamento. Em 30 dos 32 itens do instrumento, mais de 90% dos participantes avaliaram a adesão ao comportamento de cuidado no nível "sempre". Por categorias, observou-se que para "Disponibilidade para o cuidado" houve significativamente menor pontuação entre os pacientes de 18 a 49 anos (p=0.0455). Na categoria "Abertura à comunicação enfermeiro-paciente" houve menor pontuação mediana nos pacientes com Mieloma (p=0.0043) e nos pacientes em fase de Remissão-Consolidação (p=0.0084). Os dias de internação estiveram significativamente associados à categoria "Disponibilidade para o atendimento", sendo menos com 16 dias e mais (p=0.0242). Conclusão. Observou-se alta frequência de comportamentos de cuidado humanizado e pequenas diferenças na avaliação que estiveram associadas a fatores demográficos como idade e a fatores clínicos como diagnóstico, fase do tratamento e dias de internação.

Concordance Between Clinical and Laboratory Diagnosis of Abnormal Vaginal Discharge in Chilean Women / Concordância entre o diagnóstico clínico e laboratorial de corrimento vaginal anormal em mulheres chilenas

Abstract Objective To determine the concordance between the clinical diagnosis of women with abnormal vaginal discharge (AVD) and laboratory results using molecular detection and observation of the vaginal microbiota. Methods Cross-sectional study conducted in 2018 in Temuco, Chile. A total of 25 midwives from 12 health centers participated. A total of 125 women>18 years old, volunteers, were recruited. The sample of the posterior vaginal fornix was obtained by speculoscopy. Characteristics of the discharge and of the external and internal genitalia were observed. Gram staining was used to observe vaginal microbiota, blastoconidia and pseudohyphae, and polymerase chain reaction was used for the detection of Trichomonas vaginalis and Candida albicans. The Cohen kappa coefficient was used in the concordance analysis. Results Out of a total of 125 women with AVD, 85.6% consulted spontaneously and 14.4% were diagnosed clinically during a routine check-up. Absolute concordance was significant (p=0.0012), with an agreement of 13.6%. The relative concordance was significant, but fair for bacterial vaginosis (Kappa=0.21; p=0.003) and candidiasis (Kappa=0.22; p=0.001), and slight for trichomoniasis (Kappa=0.14; p=0.009). The percentage of coincidence of the diagnoses (single or mixed) by laboratory and midwives was: bacterial vaginosis 63.2% (12/19), candidiasis 36.5% (27/74), and trichomoniasis 12.5% (4/32). There was 20% coinfection. A total of 36% of the clinical diagnoses of AVD had negative laboratory tests. Conclusion The vulvovaginitis conditions candidiasis and trichomoniasis appear to be overdiagnosed, and bacterial vaginosis appears to be underdiagnosed by the clinical diagnosis when compared with the laboratory diagnosis. The low concordance obtained shows the importance of complementing the clinical diagnosis with a laboratory study of AVD, particularly in women with failed treatments and/or coinfections with unspecific and varying signs and symptoms.

Resumo Objetivo Determinar a concordância entre o diagnóstico clínico de mulheres com corrimento vaginal anormal (AVD) e os resultados laboratoriais por meio da detecção molecular e observação da microbiota vaginal. Métodos Estudo transversal realizado em 2018 em Temuco, Chile. Participaram 25 parteiras de 12 centros de saúde. Um total de 125 mulheres>18 anos, voluntárias, foramrecrutadas. A amostra do fórnice vaginal posterior foi obtida por especuloscopia. Foram observadas características da secreção e da genitália externa e interna. A coloração de Gram foi usada para observar a microbiota vaginal, blastoconídios e pseudo-hifas, e a reação em cadeia da polimerase foi usada para a detecção de Trichomonas vaginalis e Candida albicans. O coeficiente kappa de Cohen foi usado na análise de concordância. Resultados De um total de 125 mulheres com AVD, 85,6% consultaram espontaneamente e 14,4% foram diagnosticados clinicamente durante um check-up de rotina. A concordância absoluta foi significativa (p=0,0012), com concordância de 13,6%. A concordância relativa foi significativa, mas razoável para vaginose bacteriana (Kappa =0,21; p=0,003) e candidíase (Kappa=0,22; p=0,001), e leve para tricomoníase (Kappa=0,14; p=0,009). O percentual de coincidência dos diagnósticos (solteiros ou mistos) por laboratório e parteiras foi: vaginose bacteriana 63,2% (12/19), candidíase 36,5% (27/74) e tricomoníase 12,5% (4/32). Houve 20% de coinfecção. Umtotal de 36% dos diagnósticos clínicos de AVD tiveram exames laboratoriais negativos. Conclusão As condições de vulvovaginite candidíase e tricomoníase parecem ser sobrediagnosticadas, e a vaginose bacteriana parece ser subdiagnosticada pelo diagnóstico clínico quando comparado com o diagnóstico laboratorial. A baixa concordância obtida mostra a importância de complementar o diagnóstico clínico comestudo laboratorial de AVD, principalmente emmulheres com falha de tratamento e / ou coinfecções com sinais e sintomas inespecíficos e variáveis.

First Molecular Characterization of Trichostrongylus colubriformis Infection in Rural Patients from Chile.

PURPOSE: The aim was to characterize the infection by Trichostrongylus spp. in patients from Chile using a combination of molecular detection techniques and phylogenetic analysis relating the findings to clinical and epidemiological reports of the patients METHODS: Strongylid eggs were detected in seven patients by coproparasitological techniques. From each sample a fragment of the ITS-2 ribosomal gene was amplified by PCR, sequenced and analyzed by the Neighbor-Joining method. RESULTS: All the sequences and phylogenetic clusters corresponded to T. colubriformis. Two samples presented a single nucleotide polymorphism showing two possible haplotypes. Six patients presented gastrointestinal symptoms. All of them lived on farms and used sheep manure as fertilizer. CONCLUSION: T. colubriformis was the strongylid involved in the infections of these Chilean patients associated with the presence of livestock and agricultural practices that favor infection by this type of nematode.

[Frequency of cervical cytological alterations and human papilloma virus in a sample of university students in Temuco, Chile]. / Frecuencia de alteraciones citológicas cervicales y virus papiloma humano en una muestra de estudiantes universitarias en Temuco, Chile.

BACKGROUND: The early onset of sexual activity can promote the development of cervical alterations and sexually transmitted infections, especially the human papillomavirus (HPV) very common in adolescents and young people. AIM: The condition of the cervix, HPV and sexual behavior in young women under 25 years of age were analyzed. METHODS: 182 university students, healthy, sexually active, 18-24 years old, without vaccine for HPV participated. Papanicolaou (Pap) test and classification of high and low risk HPV (HR and LR) were performed by real time polymerase chain reaction. The sexual behaviors were consulted in private. RESULTS: The 46.9% of Pap presented cytological alterations, non-specific inflammation/hemorrhagic (29.4%) and atypical smear (10.2%) being de most frequent. The overall frequency of HPV-HR was 24.3%, of these 67.4% presented an altered Pap. There was an association between cytological alterations and HPV (p < 0.0001) and years of sexual activity and atypical smear or cervical intraepithelial neoplasia grade I (CIN I) (p = 0.009). 11.9% of young women (21/177) presented atypical smear or CIN I, with 66.7% of cases HPV-HR. CONCLUSIONS: These findings alert the vulnerability of these young women who would have a potential risk of viral persistence, CIN and eventually cancer. It is important to emphasize counseling and prevention prior to the regular age of admission to the screening program for cervical cancer. This study was financed by the Universidad de La Frontera through Projects DI15-0047 and DI17-0123.

Frecuencia de alteraciones citológicas cervicales y virus papiloma humano en una muestra de estudiantes universitarias en Temuco, Chile / Frequency of cervical cytological alterations and human papilloma virus in a sample of university students in Temuco, Chile

Resumen Introducción: El inicio precoz de actividad sexual puede favorecer el desarrollo de alteraciones cervicales y de infecciones de transmisión sexual, en especial del virus papiloma humano (VPH) muy frecuente en adolescentes y jóvenes. Objetivo: Analizar el estado del cuello uterino, presencia del VPH y conductas sexuales en mujeres menores de 25 años. Material y Métodos: Participaron 182 estudiantes universitarias de 18-24 años, sanas, sexualmente activas y no vacunadas para VPH. Se realizó Papanicolaou (Pap) y clasificación del VPH en alto y bajo riesgo (AR y BR) mediante reacción de polimerasa en cadena en tiempo real. Las conductas sexuales fueron consultadas privadamente. Resultados: El 46,9% de los Pap presentaron alteraciones citológicas (inflamación inespecífica/hemorrágico: 29,4% y frotis atípicos (FA):10,2%). La frecuencia de los VPH-AR fue 24,3%; de éstos, 67,4% presentó un Pap alterado. Hubo asociación entre alteraciones citológicas y presencia de VPH (p < 0,0001) y años de actividad sexual y FA o neoplasia intraepitelial grado I (NIE I) (p = 0,009). El 11,9% de las jóvenes estudiadas (21/177) presentó FA o NIE I, con 66,7% de casos VPH-AR. Conclusiones: Estos hallazgos alertan la vulnerabilidad de estas jóvenes que tendrían un riesgo potencial de persistencia viral, NIE y eventualmente cáncer. Es importante enfatizar consejería y prevención previo a la edad normada de ingreso al programa de cribado para cáncer cérvico uterino en Chile.

Background: The early onset of sexual activity can promote the development of cervical alterations and sexually transmitted infections, especially the human papillomavirus (HPV) very common in adolescents and young people. Aim: The condition of the cervix, HPV and sexual behavior in young women under 25 years of age were analyzed. Methods: 182 university students, healthy, sexually active, 18-24 years old, without vaccine for HPV participated. Papanicolaou (Pap) test and classification of high and low risk HPV (HR and LR) were performed by real time polymerase chain reaction. The sexual behaviors were consulted in private. Results: The 46.9% of Pap presented cytological alterations, non-specific inflammation/hemorrhagic (29.4%) and atypical smear (10.2%) being de most frequent. The overall frequency of HPV-HR was 24.3%, of these 67.4% presented an altered Pap. There was an association between cytological alterations and HPV (p < 0.0001) and years of sexual activity and atypical smear or cervical intraepithelial neoplasia grade I (CIN I) (p = 0.009). 11.9% of young women (21/177) presented atypical smear or CIN I, with 66.7% of cases HPV-HR. Conclusions: These findings alert the vulnerability of these young women who would have a potential risk of viral persistence, CIN and eventually cancer. It is important to emphasize counseling and prevention prior to the regular age of admission to the screening program for cervical cancer. This study was financed by the Universidad de La Frontera through Projects DI15-0047 and DI17-0123.

A PCR-RFLP assay for discrimination of Echinococcus granulosus sensu stricto and Taenia spp. in dogs stool.

In order to develop a method of identification and discrimination of Echinococcus granulosus sensu stricto from faecal samples of dogs infected with taeniid eggs (Echinococcus spp., Taenia spp.), a combined strategy of Polymerase Chain Reaction (PCR) and Restriction Fragments of Lenght Polymorphisms (RFLP) was proposed. Initially, a pair of primers was designed to amplify a fragment of the 12 Subunit of ribosomal RNA gene (12SrRNA) from mitochondrial DNA. The amplified product was digested by SspI restriction enzyme, which in E. granulosus kept the intact fragment of 160 basis pairs (bp), while in Taenia spp. produced two fragments (62 bp and another of 98 bp). The method was tested using positive controls of DNA, in faecal samples experimentally contaminated with eggs of E. granulosus and Taenia spp. and in dogs naturally infected. In all of them, reproducible results were obtained and the primers were specific to amplify only Taeniidae DNA. The sensitivity of the technique was tested, achieving amplification of DNA extractions with a single egg. In conclusion, the technique developed was optimal and easy to identify patent infections by E. granulosus s.s., constituting a possible alternative for epidemiological studies in dogs, especially in endemic areas where this infection occurs.

Trichinella spiralis Infecting Wild Boars in Southern Chile: Evidence of an Underrated Risk.

Trichinellosis in Chilean wild animals has scarcely been documented. The introduction of wild boars into the wild environment represents a viable new host with a potential risk of infection for human health. The aim of this study was to determine the presence and prevalence of Trichinella in wild boars. Two hundred seventy eight wild boars from of the Southern Chile were examined by compression and artificial digestion techniques. The larvae in the positive samples were collected for taxonomic analysis through polymerase chain reaction-inter-simple sequence repeats and to calculate the parasitic burden. A prevalence of 1.8% (5/278) of infected animals and an average parasitic burden of 6.8 ± 2.1 larvae per gram were estimated. The only species identified by molecular techniques was Trichinella spiralis. Prevalence of T. spiralis in wild boars was similar to those described around the world. T. spiralis infection rate and parasite burden detected in Chilean wild boars represent a certain food-borne risk for human population.

DNA extraction in Echinococcus granulosus and Taenia spp. eggs in dogs stool samples applying thermal shock.

The extraction of DNA in taeniid eggs shows complications attached to the composition of stool samples and the high resistance of eggs to degradation. The objective of this study was to test a method of DNA extraction in taeniid eggs by applying a thermal shock to facilitate the chemical-enzymatic degradation of these elements. A group of six tubes containing 1 ml of dog stool sample was spiked with eggs of Echinococcus granulosus and another group of six with Taenia pisiformis. Samples were floated with supersaturated sugar solution and centrifuged. The upper portion of each tube (500 µl) was aspirated and deposited in 1.5 ml tubes. Three tubes from each group were incubated at -20 °C and then at 90 °C, the remaining three from each group, incubated at room temperature. Proteinase K and lysis buffer were added to each tube and incubated for 12 h at 58 °C. The lysis effect was evaluated by microscopy at 3, 6 and 12 h and integrity by electrophoresis in 1% agarose gels. With the same experimental scheme, the thermal shock effect was evaluated in extractions of 1, 2, 3 and 4 eggs of each species and the DNA was quantified. Additionally, the protocol was applied in samples of 4 dogs diagnosed with natural infection by Taeniidae worms. Finally, all the extractions were tested by PCR amplification. Both E. granulosus and T. pisiformis eggs showed a similar response in the tests. In samples without treatment, the lysis effect was poor and showed no differences over time, but in those subjected to thermal shock, eggs degradation increased with time. In both treatments, there was no DNA loss integrity. The protocol applied to limited amounts of eggs yielded PCR products in 100% of the samples exposed to thermal shock, allowing PCR amplifications up to 1 egg. In non-exposed samples, the results were not replicable. However, DNA quantification showed low values in both treatments. In turn, DNA extractions with thermal shock in infected dog samples finally yielded PCR amplifications in 100%. It was concluded that thermal shock facilitates the DNA extraction for molecular analysis in taeniid eggs. The technique is effective extracting DNA even from a single egg and also to analyze natural infections samples with a relatively simple implementation.

[Human papilloma virus and Chlamydia trachomatis by number of sexual partners and time of sexual activity on university students in the Region of La Araucanía, Chile]. / Virus papiloma humano y Chlamydia trachomatis según número de parejas sexuales y tiempo de actividad sexual en estudiantes universitarias en la Región de La Araucanía, Chile.

BACKGROUND: Human papilloma virus (HPV) and Chlamydia trachomatis are the most prevalent sexually transmitted infections (STIs), among teenagers and young people, with risk factors: active sex life and multiple partners. Chlamydia trachomatis infection may favor HPV infection and this, the development of cervical cancer. Both infections can lead to consequences on sexual and reproductive health. OBJECTIVE: To determine frequency of HPV and C. trachomatis in asymptomatic university women less than 25 years, associating them with number of sexual partners (n°SxP) and time of sexual activity (TSxA). Material andMethods: 151 cervical samples for HPV and C. trachomatis, were processed by conventional and in real time reaction polymerase chain. RESULTS: HPV 21, 8%, C. trachomatis 11, 2% and co-infection (HPV/C.trachomatis), 4.6%. Aimong HPV +, 80, 6% showed high risk HPV. The n°SxP was strongly associated with HPV. Aimong young coinfected HPV/C. trachomatis, 71.4% had 3 or more PSx. Chlamydia trachomatis was more frequent (64,7%) that HPV within range of 3-5 years according to the TSxA, Discussion: A high prevalence of HPV and C. trachomatis was observed. Young women with coinfection HPV/C. trachomatis could be a high-risk group need to monitor their infections. It suggests the implementation of university programs in education, counseling and prevention in sexual health.

Virus papiloma humano y Chlamydia trachomatis según número de parejas sexuales y tiempo de actividad sexual en estudiantes universitarias en la Región de La Araucanía, Chile / Human papilloma virus and Chlamydia trachomatis by number of sexual partners and time of sexual activity on university students in the Region of La Araucanía, Chile

Introducción: El virus papiloma humano (VPH) y Chlamydia trachomatis son las infecciones de transmisión sexual (ITS) más frecuentes, en adolescentes y jóvenes, con factores de riesgo: vida sexual activa y múltiples parejas. Chlamydia trachomatis puede favorecer el ingreso de VPH y éste el desarrollo del cáncer cérvico uterino. Ambas infecciones pueden dejar secuelas en la salud sexual y reproductiva. Objetivo: Determinar frecuencias VPH y C trachomatis en estudiantes universitarias asinto-máticas bajo 25 años, asociándolas con número de parejas sexuales (n°PSx) y tiempo de actividad sexual (TASx). Material y Método: Se procesaron 151 muestras/exo y endo cervicales para VPH y C. trachomatis, mediante reacción de polimerasa en cadena convencional y en tiempo real. Resultados: La frecuencia fue: VPH 21,8%, C. trachomatis 11,2% y co-infección 4,6%. De las jóvenes con infección por VPH, 80,6% presentó VPH alto riesgo oncogénico. El n°PSx se asoció fuertemente a VPH. Entre las jóvenes con co-infección VPH/C. trachomatis, 71,4% tenían tres o más PSx. Según TASx, C. trachomatis fue más frecuente (64,7%) entre 3-5 años que VPH. Conclusión: Se observó alta prevalencia de VPH y C trachomatis. Mujeres jóvenes con co-infección VPH/C. trachomatis podrían ser un grupo de alto riesgo con necesidad de mo-nitorear sus infecciones. Es sugerida la implementación de programas universitarios en educación, orientación y prevención en ITS.

Background: Human papilloma virus (HPV) and Chlamydia trachomatis are the most prevalent sexually transmitted infections (STIs), among teenagers and young people, with risk factors: active sex life and multiple partners. Chlamydia trachomatis infection may favor HPV infection and this, the development of cervical cancer. Both infections can lead to consequences on sexual and reproductive health. Objective: To determine frequency of HPV and C. trachomatis in asymptomatic university women less than 25 years, associating them with number of sexual partners (n°SxP) and time of sexual activity (TSxA). Material andMethods: 151 cervical samples for HPV and C. trachomatis, were processed by conventional and in real time reaction polymerase chain. Results: HPV 21, 8%, C. trachomatis 11, 2% and co-infection (HPV/C.trachomatis), 4.6%. Aimong HPV +, 80, 6% showed high risk HPV. The n°SxP was strongly associated with HPV. Aimong young coinfected HPV/C. trachomatis, 71.4% had 3 or more PSx. Chlamydia trachomatis was more frequent (64,7%) that HPV within range of 3-5 years according to the TSxA, Discussion: A high prevalence of HPV and C. trachomatis was observed. Young women with coinfection HPV/C. trachomatis could be a high-risk group need to monitor their infections. It suggests the implementation of university programs in education, counseling and prevention in sexual health.

Oestrus ovis infection of grazing sheep during summer in southern Chile

Oestrus ovis is a botfly whose larvae cause nasal myiasis, an environmental-dependent disease in small ruminants, generating acute and chronic injuries in the cranial cavities of sheep. Chile is a country of the southernmost worldwide distribution of this parasite, and there is few information about. Whence, the objective of this study was to approximate the epidemiological situation of O. ovis infection in Chilean sheep. From December 2009 to March 2010, a total of 87 samples were obtained by necropsy for skull inspection. The larvae were collected, and microscopically identified. The prevalence in the sampled sheep was 60.9%. From those that were infected, 85.7% (18/21) of sheep were 1 to 3 years old, constituting the stratum with the highest prevalence. The difference of infection in females and males was not significant. The high risk of infection seems to be dependent upon the environmental conditions of this southern region, especially during summer when the first larval stage (L1) could be found as the evidence. Therefore, the disease should be considered as a significant problem for this kind of livestock production.

Genetic heterogeneity in internal transcribed spacer genes of Balantidium coli (Litostomatea, Ciliophora).

The species Balantidium coli is the only ciliate that parasitizes humans. It has been described in other primates, and it has been proposed that the species B. suis from pigs and B. struthionis from ostriches are synonyms of B. coli. Previous genetic analysis of pig and ostrich Balantidium isolates found a genetic polymorphism in the ITS region but its taxonomic relevance was not established. We have extended the genetic analysis to Balantidium isolates of pig, gorilla, human and ostrich origin. We have PCR-amplified and sequenced the ITS region of individual Balantidium cells. The predicted ITS secondary structures of the sequences obtained were transferred by homology modelling to the sequences of other Trichostomatia ciliates (Isotricha, Troglodytella, Lacrymaria and Spathidium) and compared to determine the importance of the differences in the primary sequences. The results show that the ITS2 secondary structure of the species considered follows the general pattern of other ciliates, although with some deviations. There are at least two main types of ITS sequence variants in B. coli which could be present in the same cell and they are common to the mammal and avian hosts studied. These data do not support B. suis and B. struthionis as distinct species.