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

Adriana Higuera; Ximena Villamizar; Giovanny Herrera; Julio Cesar Giraldo; Luis Reinel Vasquez-A; Plutarco Urbano; Oswaldo Villalobos; Catalina Tovar; Juan David Ramírez

doi:10.7717/peerj.8554

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

PeerJ. 2020 Mar 9:8:e8554. doi: 10.7717/peerj.8554. eCollection 2020.

Authors

Adriana Higuera¹, Ximena Villamizar¹, Giovanny Herrera¹, Julio Cesar Giraldo², Luis Reinel Vasquez-A³, Plutarco Urbano⁴, Oswaldo Villalobos⁵, Catalina Tovar⁶, Juan David Ramírez¹

Affiliations

¹ Grupo de Investigaciones Microbiológicas-UR (GIMUR), Departamento de Biología, Facultad de Ciencias Naturales, Universidad del Rosario, Bogota, Colombia.
² Programa de Biología, Universidad INCCA de Colombia, Bogota, Colombia.
³ Centro de Estudios en Microbiología y Parasitología (CEMPA), Departamento de Medicina Interna, Facultad de Ciencias de la Salud, Universidad del Cauca, Popayan, Colombia.
⁴ Grupo de Investigaciones Biológicas de la Orinoquia, Unitrópico, Yopal, Colombia.
⁵ Hospital Local Santa María de Mompox, Programas Especiales (Lepra y TB), Mompox, Bolivar, Colombia.
⁶ Grupo de Enfermedades Tropicales y Resistencia Bacteriana, Universidad del Sinú, Monteria, 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.

Keywords: Blastocystis; Cryptosporidium; Entamoeba; Giardia intestinalis; Molecular genotyping.

Grants and funding

Adriana Higuera was financed by the Departamento Administrativo de Ciencia, Tecnología e Innovación (Colciencias) within the framework of the National Program for Promoting Research Training (sponsorship call 757). This work was funded by the Departamento Administrativo de Ciencia, Tecnología e Innovación (Colciencias) through project “Desarrollo de una estrategia y plataforma de Multilocus Sequence Typing (MLST) para la caracterización molecular de siete parásitos intestinales,” code 122271250521 and contract number 130–2016. Funding was also received from the Vicerrectoria de Investigaciones of the Universidad del Cauca (code 4204). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.