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      Detection and genotypes of Toxoplasma gondii DNA in feces of domestic cats in Colombia Translated title: Détection d’ADN et génotypes de Toxoplasma gondii dans les fèces de chats domestiques en Colombie

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          Abstract

          The high prevalence of Toxoplasma gondii in the human population in Colombia has been linked to the existence of a high density of urban stray cats, exposing the whole population to a high density of oocysts. The goal of this study was to determine the DNA prevalence of T. gondii by conventional PCR and to phylogenetically analyze ROP18 sequences from positive samples in domestic cat ( Felis catus) fecal samples in the city of Armenia, Quindío. Fecal samples from 140 cats were collected from 10 districts around the city. Samples were concentrated using Ritchie’s method and analyzed through optical microscopy. Concentrates were used for DNA extraction followed by nested PCR amplification for T. gondii gene B1. PCR for ROP18 was performed on all B1 positive samples; the ROP18 sequences obtained were related to the Archetype I Brazilian and Chinese strains. No oocysts were detected by optical microscopy; however, 17.8% (25/140) B1 and 24% (6/25) ROP18 PCR-positive samples were detected. Phylogenetic analyses showed that isolates clustered into a single group. We assessed whether associations existed between T. gondii positive fecal samples and survey variables such as cat healthcare and socioeconomic characteristics of owners, but no statistically significant associations were found. The presence of T. gondii in cat feces is an important factor contributing to the high prevalence in the human population of this city.

          Translated abstract

          La forte prévalence de Toxoplasma gondii dans la population humaine en Colombie a été liée à l’existence d’une forte densité de chats errants urbains, exposant l’ensemble de la population à une forte densité d’oocystes. Le but de ce travail était de déterminer la prévalence de l’ADN de T. gondii par PCR conventionnelle et d’analyser phylogénétiquement les séquences ROP18 d’échantillons positifs dans des échantillons fécaux de chat domestique ( Felis catus) dans la ville d’Armenia, Quindío. Des échantillons fécaux de 140 chats ont été collectés dans 10 districts de la ville. Les échantillons ont été concentrés en utilisant la méthode de Ritchie et analysés par microscopie optique. Des concentrés ont été utilisés pour l’extraction d’ADN suivie d’une amplification par PCR nichée pour le gène B1 de T. gondii. La PCR pour ROP18 a été réalisée sur tous les échantillons positifs pour B1 ; les séquences ROP18 obtenues étaient apparentées aux souches Archétype I brésiliennes et chinoises. Aucun oocyste n’a été détecté par microscopie optique mais les échantillons étaient positifs par PCR pour 17,8 % (25/140) pour B1 et 24 % (6/25) pour ROP18. Les analyses phylogénétiques ont montré que les isolats formaient un seul groupe. Nous avons évalué s’il existait des associations entre des échantillons fécaux positifs à T. gondii et des variables d’enquête telles que les soins de santé des chats et les caractéristiques socioéconomiques des propriétaires, mais aucune association statistiquement significative n’a été trouvée. La présence de T. gondii dans les excréments de chats est un facteur important contribuant à la forte prévalence dans la population humaine de cette ville.

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          Most cited references 37

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          An ether sedimentation technique for routine stool examinations.

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            History of the discovery of the life cycle of Toxoplasma gondii.

             Vinod Dubey (2009)
            It has been 100 years since the discovery of Toxoplasma gondii in 1908. Its full life cycle was not discovered until 1970 when it was found that it is a coccidian parasite of cats with all non-feline warm blooded animals (including humans) as intermediate hosts. The discovery of the environmentally resistant stage of the parasite, the oocyst, made it possible to explain its worldwide prevalence. In the present paper, events associated with the discovery of its life cycle are recalled.
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              Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction.

              We applied the polymerase chain reaction to detection of the pathogenic protozoan Toxoplasma gondii based on our identification of a 35-fold-repetitive gene (the B1 gene) as a target. Using this procedure, we were able to amplify and detect the DNA of a single organism directly from a crude cell lysate. This level of sensitivity also allowed us to detect the B1 gene from purified DNA samples containing as few as 10 parasites in the presence of 100,000 human leukocytes. This is representative of the maximal cellular infiltration (10(5)/ml) in 1 ml of cerebrospinal fluid obtained from patients with toxoplasmic encephalitis. The B1 gene is present and conserved in all six T. gondii strains tested to date, including two isolates from patients with acquired immunodeficiency syndrome. No signal was detected by using this assay and DNAs from a variety of other organisms, including several which might be found in the central nervous system of an immunocompromised host. This combination of sensitivity and specificity should make detection of the B1 gene based on polymerase chain reaction amplification a very useful method for diagnosis of toxoplasmosis both in immunocompromised hosts and in congenitally infected fetuses.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2020
                17 April 2020
                : 27
                : ( publisher-idID: parasite/2020/01 )
                Affiliations
                [1 ] Grupo de Estudio en Parasitología y Micología Molecular (GEPAMOL), Centro de Investigaciones Biomédicas, Facultad Ciencias de la Salud, Universidad del Quindío 630004 Armenia Colombia
                [2 ] Grupo de Biodiversidad y Conservación Genética, Instituto de Genética, Universidad Nacional de Colombia 111321 Bogotá Colombia
                Author notes
                [* ]Corresponding author: oazamorav@ 123456uqvirtual.edu.co
                Article
                parasite190097 10.1051/parasite/2020023
                10.1051/parasite/2020023
                7164795
                32302287
                © A. Zamora-Vélez et al., published by EDP Sciences, 2020

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 3, Tables: 3, Equations: 0, References: 45, Pages: 7
                Categories
                Research Article

                rop18, toxoplasma gondii, cats, pcr, prevalence

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