Multilocus Sequence Typing helps understand the genetic diversity of Cryptosporidium hominis and Cryptosporidium parvum isolated from Colombian patients.

Multilocus Sequence Typing has become a useful tool for the study of the genetic diversity and population structure of different organisms. In this study, a MLST approach with seven loci (CP47, MS5, MS9, MSC6-7, TP14, and gp60) was used to analyze the genetic diversity of Cryptosporidium hominis and Cryptosporidium parvum isolated from 28 Colombian patients. Five Cryptosporidium species were identified: C. hominis, C. parvum, Cryptosporidium felis, Cryptosporidium meleagridis, and Cryptosporidium suis. Unilocus gp60 analysis identified four allelic families for C. hominis (Ia, Ib, Id, and Ie) and two for C. parvum (IIa and IIc). There was polymorphic behavior of all markers evaluated for both C. hominis and C. parvum, particularly with the CP47, MS5, and gp60 markers. Phylogenetic analysis with consensus sequences (CS) of the markers showed a taxonomic agreement with the results obtained with the 18S rRNA and gp60 gene. Additionally, two monophyletic clades that clustered the species C. hominis and C. parvum were detected, with a higher number of subclades within the monophyletic groups compared to those with the gp60 gene. Thirteen MLG were identified for C. hominis and eight for C. parvum. Haplotypic and nucleotide diversity were detected, but only the latter was affected by the gp60 exclusion from the CS analysis. The gene fixation index showed an evolutionary closeness between the C. hominis samples and a less evolutionary closeness and greater sequence divergence in the C. parvum samples. Data obtained in this work support the implementation of MLST analysis in the study of the genetic diversity of Cryptosporidium, considering the more detailed information that it provides, which may explain some genetic events that with an unilocus approach could not be established. This is the first multilocus analysis of the intra-specific variability of Cryptosporidium from humans in South America.

Prevalence of intestinal parasites in a cohort of HIVinfected patients from Antioquia, Colombia. / Prevalencia de parásitos intestinales en una cohorte de pacientes positivos para HIV en Antioquia, Colombia

INTRODUCTION: HIV infection is still a public health problem worldwide and co-infections with other infectious agents including intestinal parasites are of particular concern, mainly in developing countries like Colombia. OBJECTIVE: To conduct a cross-sectional study in patients attending an HIV care program in Antioquia given that there have been few intestinal parasites prevalence studies among the HIV population in the country. MATERIAL AND METHODS: We evaluated stool samples from 192 patients by direct wet mount and concentration, modified Ziehl Neelsen staining, and agar plate culture. Univariate and correlation analyses were done to explore the association between socio-demographic and clinical characteristics and parasitological data. RESULTS: The overall prevalence of intestinal parasites in HIV-positive subjects was 29.2% (56/192; 95% CI: 22.8% - 35.6%). Entamoeba histolytica/dispar/moshkosvkii with 13.0% (25/192; 95% CI: 8.2% - 17.8%) and Blastocystis with 12.0% (23/192; 95% CI: 7.4% - 16.6%) were the most frequent. Opportunistic parasites like Cryptosporidium spp. and Cystoisospora belli were less prevalent, each one with 0.5% of positive samples (1/192; 95% CI: 0.1% - 1.5%). Commensal protozoa were also detected with a prevalence of 18.8% (36/192; 95% CI: 13.3% - 24.3%). Most of the individuals in the study had a controlled viral load and an LTCD4 count greater than 200 cel/µL. A small percentage (9.3%) had diarrhea. Bivariate analysis and multivariate logistic regression showed that only age and having pets had a significant association with intestinal parasites in this cohort. CONCLUSIONS: Our results confirmed that the evaluated population is at high risk of intestinal parasite infection, which highlights the need for routine screening of gastrointestinal parasites to provide prompt treatment and reduce possible complications.

Introducción. La infección por HIV y las coinfecciones con otros agentes infecciosos, incluidos los parásitos intestinales, son motivo de especial preocupación en países en desarrollo como Colombia. Objetivo. Hacer un estudio transversal en pacientes que asisten a un programa de atención de HIV en el departamento de Antioquia, dado que los estudios de prevalencia de parásitos intestinales en la población con HIV son escasos en el país. Materiales y métodos. Se evaluaron 192 muestras de materia fecal mediante examen coprológico directo y por concentración, tinción de Ziehl-Neelsen modificada, y aislamiento en agar. Se hicieron análisis univariados y de correlación, para explorar la asociación entre las características sociodemográficas y clínicas, y los datos parasitológicos. Resultados. La prevalencia global de parásitos intestinales en pacientes positivos para VIH fue del 29.2 % (56/192; IC95% 22.8-35.6 %), siendo Entamoeba histolytica/dispar/moshkosvkii, con 13.0 % (25/192; IC95% 8.2-17.8 %), y Blastocystis, con 12.0 % (23/192; IC95% 7.4-16.6 %), los mas frecuentes. Los parásitos oportunistas Cryptosporidium spp. y Cystoisospora belli fueron menos prevalentes, cada uno con 0.5 % (1/192; IC95% 0.1-1.5 %) de muestras positivas. También, se detectaron protozoos comensales, con una prevalencia del 18,8 % (36/192; IC95% 13,3-24,3 %). La mayoría de los individuos tenía una carga viral controlada y un recuento de linfocitos T CD4 superior a 200 células/µl. Un pequeño porcentaje (9,3 %) presentó diarrea. La edad y el tener mascotas mostraron una asociación significativa con la presencia de parásitos intestinales. Conclusión. Se confirmó que la población evaluada tiene un alto riesgo de infección por parásitos intestinales, lo que resalta la necesidad de un protocolo de diagnóstico para el cribado de dichos agentes, con el fin de brindar un tratamiento rápido y reducir las posibles complicaciones.

Prevalence of intestinal parasites in a cohort of HIV- infected patients from Antioquia, Colombia / Prevalencia de parásitos intestinales en una cohorte de pacientes positivos para HIV en Antioquia, Colombia

Abstract | Introduction: HIV infection is still a public health problem worldwide and co-infections with other infectious agents including intestinal parasites are of particular concern, mainly in developing countries like Colombia. Objective: To conduct a cross-sectional study in patients attending an HIV care program in Antioquia given that there have been few intestinal parasites prevalence studies among the HIV population in the country. Material and methods: We evaluated stool samples from 192 patients by direct wet mount and concentration, modified Ziehl Neelsen staining, and agar plate culture. Univariate and correlation analyses were done to explore the association between socio-demographic and clinical characteristics and parasitological data. Results: The overall prevalence of intestinal parasites in HIV-positive subjects was 29.2% (56/192; 95% CI: 22.8% - 35.6%). Entamoeba histolytica/dispar/moshkosvkii with 13.0% (25/192; 95% CI: 8.2% - 17.8%) and Blastocystis with 12.0% (23/192; 95% CI: 7.4% -16.6%) were the most frequent. Opportunistic parasites like Cryptosporidium spp. and Cystoisospora belli were less prevalent, each one with 0.5% of positive samples (1/192; 95% CI: 0.1% - 1.5%). Commensal protozoa were also detected with a prevalence of 18.8% (36/192; 95% CI: 13.3% - 24.3%). Most of the individuals in the study had a controlled viral load and an LTCD4 count greater than 200 cel/µL. A small percentage (9.3%) had diarrhea. Bivariate analysis and multivariate logistic regression showed that only age and having pets had a significant association with intestinal parasites in this cohort. Conclusions: Our results confirmed that the evaluated population is at high risk of intestinal parasite infection, which highlights the need for routine screening of gastrointestinal parasites to provide prompt treatment and reduce possible complications.

Resumen | Introducción. La infección por HIV y las coinfecciones con otros agentes infecciosos, incluidos los parásitos intestinales, son motivo de especial preocupación en países en desarrollo como Colombia. Objetivo. Hacer un estudio transversal en pacientes que asisten a un programa de atención de HIV en el departamento de Antioquia, dado que los estudios de prevalencia de parásitos intestinales en la población con HIV son escasos en el país. Materiales y métodos. Se evaluaron 192 muestras de materia fecal mediante examen coprológico directo y por concentración, tinción de Ziehl-Neelsen modificada, y aislamiento en agar. Se hicieron análisis univariados y de correlación, para explorar la asociación entre las características sociodemográficas y clínicas, y los datos parasitológicos. Resultados. La prevalencia global de parásitos intestinales en pacientes positivos para VIH fue del 29.2 % (56/192; IC95% 22.8-35.6 %), siendo Entamoeba histolytica/dispar/moshkosvkii, con 13.0 % (25/192; IC95% 8.2-17.8 %), y Blastocystis, con 12.0 % (23/192; IC95% 7.4-16.6 %), los mas frecuentes. Los parásitos oportunistas Cryptosporidium spp. Y Cystoisospora belli fueron menos prevalentes, cada uno con 0.5 % (1/192; IC95% 0.1-1.5 %) de muestras positivas. También, se detectaron protozoos comensales, con una prevalencia del 18,8 % (36/192; IC95% 13,3-24,3 %). La mayoría de los individuos tenía una carga viral controlada y un recuento de linfocitos T CD4 superior a 200 células/μl. Un pequeño porcentaje (9,3 %) presentó diarrea. La edad y el tener mascotas mostraron una asociación significativa con la presencia de parásitos intestinales. Conclusión. Se confirmó que la población evaluada tiene un alto riesgo de infección por parásitos intestinales, lo que resalta la necesidad de un protocolo de diagnóstico para el cribado de dichos agentes, con el fin de brindar un tratamiento rápido y reducir las posibles complicaciones.

Common occurrence of Cryptosporidium hominis in children attending day-care centers in Medellin, Colombia.

Cryptosporidium spp. are apicomplexan protozoa associated with chronic diarrhea in AIDS and other immunocompromised patients, and one of the commonest causes of childhood diarrhea and malnutrition, particularly in low-income settings. In Colombia, there are few molecular epidemiological studies on Cryptosporidium spp.; thereby, the transmission dynamics of this parasite in the country is poorly known. This study evaluated the diversity of Cryptosporidium at species, subtype family, and subtype level in children attending various day-care centers in Medellin, Colombia. Two hundred and ninety stool samples from children < 5 years of age were collected from April to November of 2015. All samples were processed by PCR and sequence analysis of the ssu RNA gene and the gp60 gene. An infection rate of 2.4% was observed, with only two Cryptosporidium species identified: C. hominis (6/7) and C. meleagridis (1/7). Cryptosporidium hominis isolates belonged to the subtypes IbA10G2, IaA13R6 and IaA13R7; IIIbA26G1R1 C. meleagridis subtype was also detected. There is a C. hominis predominance in the children evaluated, suggesting an important role of the anthroponotic transmission cycle in the day-care centers analyzed. Further investigation is required to determine infection sources and susceptible hosts in order to define appropriate management of cryptosporidiosis.