Molecular and descriptive epidemiology of intestinal protozoan parasites of children and their pets in Cauca, Colombia: a cross-sectional study.

BACKGROUND: Parasitic infections, particularly those caused by protozoa, represent a considerable public health problem in developing countries. Blastocystis, Giardia duodenalis, Cryptosporidium spp. and the Entamoeba complex (Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii) are the most common etiological causes of intestinal parasitic infections. METHODS: We carried out a descriptive cross-sectional study in school-age children attending a daycare institution in commune eight of Popayán, Cauca (Southwest Colombia). A total of 266 fecal samples were collected (258 from children and eight from pets). Blastocystis, G. duodenalis, Cryptosporidium spp. and the Entamoeba complex were identified by microscopy, quantitative real-time PCR (qPCR) and conventional PCR. The concordance of qPCR and microscopy was assessed using the Kappa index. Molecular characterization was conducted to identify Blastocystis subtypes (18S), G. duodenalis assemblages (tpi and gdh) and Cryptosporidium species/subtypes (18S and GP60). Potential associations between intestinal parasitism and sociodemographic factors were examined using bivariate analyses. RESULTS: A total of 258 fecal samples from children were analyzed by microscopy and 255 samples were analyzed by qPCR. The prevalence of Blastocystis was between 25.19% (microscopy) and 39.22% (qPCR), that of G. duodenalis was between 8.14% (microscopy) and 10.59% (qPCR), that of Cryptosporidium spp. was estimated at 9.8% (qPCR), and that of the Entamoeba complex was between 0.39% (conventional PCR) and 0.78% (microscopy). The concordance between microscopy and qPCR was very low. Blastocystis ST1 (alleles 4, 8, and 80), ST2 (alleles 11, 12, and 15), ST3 (alleles 31, 34, 36, 38,57, and 151), and ST4 (alleles 42 and 91), G. duodenalis assemblages AII, BIII, BIV and D, C. parvum subtype IIa and C. hominis subtype IbA9G3R2 were identified. The only identified member of the Entamoeba complex corresponded to E. histolytica. No statistically significant association was identified between parasitic infection and any sociodemographic variable. CONCLUSION: This study revealed the usefulness of molecular methods to depict the transmission dynamics of parasitic protozoa in southwest Colombia. The presence of some of these protozoa in domestic animals may be involved in their transmission.

First isolation and molecular characterization of Cryptococcus neoformans var. grubii in excreta of birds in the urban perimeter of the Municipality of Popayán, Colombia.

BACKGROUND: Cryptococcosis is a systemic opportunistic mycosis, caused by Cryptococcus neoformans and Cryptococcus gattii, which affects mainly the central nervous system of immunocompromised patients; no reports have been made on the isolation of the fungus from the environment of Popayán, Colombia. AIMS: The main objective of this investigation was to determinate the incidence of C. neoformans in the urban perimeter in the City of Popayán, Colombia. METHODS: A total of 303 samples from droppings of Columba livia and Bubulcus ibis were collected between September 2012 and June 2013. The samples were processed by conventional techniques; identification of colonies was performed by biochemical tests, and molecular patterns were determined by PCR fingerprinting with the primer (GTG)5 and restriction fragment length polymorphism (RFLP) of the gene URA5. RESULTS: A total of 118 (38.94%) samples were positive for Cryptococcus in excreta of C. livia, and 361 strains belonging to Cryptococcus neoformans var. grubii were isolated. From the latter, 99.2% corresponded to the molecular pattern VNI and 0.8% to VNII, with an increased occurrence (24.4%) at a temperature of 22.5°C and a humidity of 60.8%. The excreta of B. ibis did not show the presence of the fungus. CONCLUSIONS: C. livia excreta is a key environmental niche for C. neoformans var. grubii, type VNI, supporting growth and reproduction, and serving as a major source of infection for susceptible populations in Popayán. This represents the first report on the isolation of the agent of cryptococcosis from the environment in this region, with a significant prevalence in bird excreta.

First isolation and molecular characterization of Cryptococcus neoformans var. grubii in excreta of birds in the urban perimeter of the Municipality of Popayán, Colombia / Primer aislamiento y caracterización molecular de Cryptococcus neoformans var. grubii a partir de excrementos de aves en el perímetro urbano del municipio de Popayán, Colombia

Background: Cryptococcosis is a systemic opportunistic mycosis, caused by Cryptococcus neoformans and Cryptococcus gattii, which affects mainly the central nervous system of immunocompromised patients; no reports have been made on the isolation of the fungus from the environment of Popayán, Colombia. Aims: The main objective of this investigation was to determinate the incidence of C. neoformans in the urban perimeter in the City of Popayán, Colombia. Methods: A total of 303 samples from droppings of Columba livia and Bubulcus ibis were collected between September 2012 and June 2013. The samples were processed by conventional techniques; identification of colonies was performed by biochemical tests, and molecular patterns were determined by PCR fingerprinting with the primer (GTG)5 and restriction fragment length polymorphism (RFLP) of the gene URA5. Results: A total of 118 (38.94%) samples were positive for Cryptococcus in excreta of C. livia, and 361 strains belonging to Cryptococcus neoformans var. grubii were isolated. From the latter, 99.2% corresponded to the molecular pattern VNI and 0.8% to VNII, with an increased occurrence (24.4%) at a temperature of 22.5°C and a humidity of 60.8%. The excreta of B. ibis did not show the presence of the fungus. Conclusions: C. livia excreta is a key environmental niche for C. neoformans var. grubii, type VNI, supporting growth and reproduction, and serving as a major source of infection for susceptible populations in Popayán. This represents the first report on the isolation of the agent of cryptococcosis from the environment in this region, with a significant prevalence in bird excreta

Antecedentes: La criptococosis es una micosis sistémica oportunista provocada por Cryptococcus neoformans y Cryptococcus gattii; afecta principalmente al sistema nervioso central de pacientes inmunodeficientes. No existen publicaciones sobre el aislamiento del hongo en la región de Popayán, Colombia. Objetivos: El objetivo principal de este estudio fue establecer la incidencia de C. neoformans en el perímetro urbano de la ciudad de Popayán, Colombia. Métodos: Se recogió un total de 303 muestras de materia fecal de las aves Columba livia y Bubulcus ibis entre septiembre de 2012 y junio de 2013. Las muestras se procesaron con técnicas convencionales de cultivo; la identificación de las colonias se realizó mediante pruebas bioquímicas, y los patrones moleculares se establecieron mediante PCR con el cebador (GTG)5 y la técnica de polimorfismos de longitud de los fragmentos de restricción (RFLP, por su abreviatura en inglés) del gen URA5. Resultados: Un total de 118 (38,94%) muestras de excrementos de C. livia fueron positivas para Cryptococcus y se recuperaron 361 aislamientos pertenecientes a Cryptococcus neoformans var. grubii, de los cuales el 99,2% correspondían al patrón molecular VNI y el 0,8% al VNII, con un aumento en la incidencia (24,4%) a una temperatura de 22,5°C y una humedad del 60,8%. No hubo crecimiento de este hongo en los excrementos de B. ibis. Conclusiones: Los excrementos de C. livia son un nicho ambiental fundamental para C. neoformans var. grubii, tipo VNI; ello permite su crecimiento y reproducción, y que sean una potencial fuente de infección para la población susceptible de Popayán. Esta es la primera publicación sobre el aislamiento del agente etiológico de la criptococosis en el entorno de esta región, con una prevalencia considerable en los excrementos de aves