Molecular detection and genotyping of intestinal protozoa from different biogeographical regions of Colombia

Abstract

Background: Intestinal parasitic protozoa represent a serious problem of public health particularly in developing countries. Protozoa such as Blastocystis, Giardia intestinalis, Entamoeba histolytica and Cryptosporidium spp. are associated with diarrheal symptoms. In Colombia, there is little region-specific data on the frequency and circulating genotypes/species of these microorganisms. Therefore, the main objective of our study was to employ molecular detection and genotyping of G. intestinalis and Blastocystis, Cryptosporidium and Entamoeba spp. in samples from different biogeographical regions of Colombia. Methods: We collected 649 human fecal samples from five biogeographical regions of Colombia: the Amazon, Andean, Caribbean, Orinoco and Pacific regions. Blastocystis, G. intestinalis, Cryptosporidium spp. and Entamoeba complex were detected by microscopy and conventional PCR. Molecular genotyping was conducted to identify Blastocystis subtypes (STs) (18s), G. intestinalis assemblages (triose phosphate isomerase and glutamate dehydrogenase) and Cryptosporidium species (18s). Genetic diversity indices were determined using dnasp.5. Results: We detected G. intestinalis in 45.4% (n = 280) of samples, Blastocystis in 54.5% (n = 336) of samples, Cryptosporidium spp. in 7.3% (n = 45) of samples, Entamoeba dispar in 1.5% (n = 9) of samples, and Entamoeba moshkovskii in 0.32% (n = 2) of samples. Blastocystis STs 1-4, 8 and 9 and G. intestinalis assemblages AII, BIII, BIV, D and G were identified. The following Cryptosporidium species were identified: C. hominis, C. parvum, C. bovis, C. andersoni, C. muris, C. ubiquitum and C. felis. The Caribbean region had the highest frequency for each of the microorganisms evaluated (91.9% for G. duodenalis, 97.3% for Blastocystis, 10.8% for Cryptosporidium spp., 13.5% for E. dispar and 2.7% for E. moshkovskii). The Orinoco region had a high frequency of Blastocystis (97.2%) and the Andean region had a high frequency of G. intestinalis (69.4%). High and active transmission was apparent in several regions of the country, implying that mechanisms for prevention and control of intestinal parasitosis in different parts of the country must be improved. . 2014. Blastocystis sp. frequency and sources among children from 0 to 5 years of age attending public day care centers in Calarca. A novel, multi-parallel, real-time polymerase chain reaction approach for eight gastrointestinal parasites provides improved diagnostic capabilities to resource-limited at-risk populations.

Urban versus rural prevalence of intestinal parasites using multi-parallel qPCR in Colombia

“…16 Moreover, polyparasitism involving both STH and protozoa serves as an important indicator of inadequate sanitary conditions and continual reinfection. 9 Findings from this study suggest that children from urban slums of Medellín and rural areas of Unguía in Colombia live in highly contaminated environments with continuous exposure to intestinal parasites in these different communities. The peripheral neighborhoods of the urban poor in cities such as Medellín, Colombia, typically have informal housing development with poor housing standards, overcrowding, and inadequate infrastructure for sanitation and access to clean water.…”

“…6 In Colombia, studies reporting molecular epidemiologic data on intestinal parasites among children from both urban and rural settings in the same study population are sparse. 3,[7][8][9] Limited molecular epidemiologic surveillance data on childhood intestinal parasitism in areas with concentrations of poverty undermines positive child health outcomes. 3,9 Our study contributes to data comparing the prevalence of intestinal parasites detected by qPCR among urban slum and rural children from the study sampling areas in Colombia.…”

“…3,[7][8][9] Limited molecular epidemiologic surveillance data on childhood intestinal parasitism in areas with concentrations of poverty undermines positive child health outcomes. 3,9 Our study contributes to data comparing the prevalence of intestinal parasites detected by qPCR among urban slum and rural children from the study sampling areas in Colombia. Because the interplay between socioeconomic and environmental factors, hygiene, and transmission dynamics contribute to increased child vulnerability to intestinal parasite exposure.…”

Development of a Multilocus Sequence Typing Scheme for Giardia intestinalis

“…We randomly chose 24 samples positive for G. intestinalis , collected in different regions of Colombia, from Amazonas [ 53 ], Casanare, Bolívar, and Córdoba, as reported elsewhere [ 54 ].…”

“…Convenience sampling of human faeces samples was performed in the departments of Amazonas, Bolívar, Casanare, and Córdoba in Colombia. The collection, extraction and typing of samples positive for G. intestinalis was performed as described in Sánchez et al [ 53 ] for samples from the Amazon and in Higuera et al [ 54 ] for samples from Córdoba, Bolívar and Casanare. A small set of 24 DNAs from G. intestinalis positive samples were taken at random to evaluate markers assessed in silico.…”