Knowledge and practices about taeniasis/cysticercosis complex within the framework of the national plan for its elimination in Colombia, 2019.

The taeniasis/cysticercosis complex (TCC) belongs to the group of neglected infectious diseases with a multifactorial transmission that includes hosts such as humans and pigs of the parasitic tapeworm (Taenia solium) and environmental factors. This study represents the first phase of the National and Cross-sectoral Plan for TCC Elimination. OBJECTIVES: To obtain data about knowledge and practices in relation to the TCC from knowledge, attitudes, and practices (KAP) surveys applied in two pilot localities for building a baseline of the potential risk factors related to identification, prevention, and control of TCC in Colombia. METHODS: A descriptive cross-sectional survey from October to November in 2019 was carried out using a structured questionnaire and random sampling by single-stage conglomerates in the municipalities of Mahates, Bolívar (n = 152) (Atlantic Region) and Mercaderes, Cauca (n = 152) (Andean Region). RESULTS: Respondents in Mahates showed moderate knowledge about TCC compared to those in Mercaderes, however, risky practices were identified in both places (consumption of pork with cysts and defecation in places other than the bathroom or latrine). Deficiency in infrastructure and failure in basic services were observed as potential risk factors for TCC transmission in both municipalities. CONCLUSION: Based on the framework of the National and Cross-sectoral Plan for TCC Elimination, adequate knowledge on identification, prevention, and control of the disease must be reinforced; cultural and ecological differences should be considered when designing communication and knowledge transmission tools. We consider that major investment should be made in improving basic services and creating sustainable modernized pig farming in Mahates and Mercaderes.

Prevalence of intestinal parasitism in preschool and school children in Colombia: Systematic review and meta-analysis.

OBJECTIVE: Intestinal parasitic infections (IPIs) are a public health challenge in developing countries such as Colombia, causing anaemia and delayed growth and development in children. We aimed to estimate the geographical and prevalence trend of IPIs in the last 30 years in school and preschool children in Colombia. METHODS: We conducted a systematic review and meta-analysis. We identified potential manuscripts through PubMed, EMBASE, Web of Science, LILACS, Scielo and Google Scholar on the IPIs prevalence in school and preschool children in Colombia. Articles included in the qualitative analysis were published between 1990 and 2020 in English or Spanish and met the inclusion criteria. Subsequently, a random-effects meta-analysis, a meta-regression and a trend analysis were performed. RESULTS: We identified 2292 articles; 109 were included in the qualitative review, and 79 articles were included in the meta-analysis. The estimated IPI prevalence was 55% (95% CI: 48-63). By age group, the prevalence in preschool children was 37% (95% CI: 26-49) and 66% (95% CI: 52-78) in schoolchildren. The prevalence by region was heterogeneous, with the Amazon being the highest (69%) and the Santanderes the lowest (28%). In the last 20 years, the prevalence of helminthiasis has decreased (from 64.66% in 1990-1995 to 22.09% in 2016-2020). CONCLUSION: The prevalence of IPIs is high (>30%) in three of the seven regions in Colombia. Biannual administration of mass deworming in schoolchildren is recommended in the Amazon region. Public policies aiming to control IPIs should be reinforced. Further prevalence studies should include Cesar, Guaviare, Vichada and Vaupés, where the epidemiology of IPIs is unknown. SUSTAINABLE DEVELOPMENT GOALS: Good health and wellbeing, clean water and sanitation, sustainable cities and communities.

Succinate dehydrogenase gene as a marker for studying Blastocystis genetic diversity.

Blastocystis has been reported as the most common eukaryotic microorganism residing in the intestines of both humans and animals, with a prevalence of up to 100% in some populations. Since this is a cryptic species, sequence polymorphism are the single strategy to analyses its genetic diversity, being traditionally used the analysis of ssu rRNA gene sequence to determine alleles and subtypes (STs) for this species. This multicopy gene has shown high diversity among different STs, making necessary to explore other genes to assess intraspecific diversity. This study evaluated the use of a novel genetic marker, succinate dehydrogenase (SDHA), for the typing and evaluation of the genetic diversity and genetic population structure of Blastocystis. In total, 375 human fecal samples were collected and subjected to PCR, subtyped using the ssu rRNA marker, and then the SDHA gene was amplified via PCR for 117 samples. We found some incongruences between tree topologies for both molecular markers. However, the clustering by ST previously established for Blastocystis was congruent in the concatenated sequence. SDHA showed lower reticulation (The origination of a lineage through the partial merging of two ancestor lineages) signals and better intra ST clustering ability. Clusters with geographical associations were observed intra ST. The genetic diversity was lower in the marker evaluated compared to that of the ssu rRNA gene (nucleotide diversity = 0.03344 and 0.16986, respectively) and the sequences analyzed showed population expansion with genetic differentiation principally among STs. The ssu rRNA gene was useful to explore interspecific diversity but together with the SDHA gene the resolution power to evaluate intra ST diversity was higher. These results showed the potential of the SDHA marker for studying the intra ST genetic diversity of Blastocystis related with geographical location and the inter ST diversity using the concatenated sequences.

Development of a Multilocus Sequence Typing Scheme for Giardia intestinalis.

Giardia intestinalis is an intestinal protozoan most commonly found in humans. It has been grouped into 8 assemblages (A-H). Markers such as the glutamate dehydrogenase gene, triose phosphate isomerase and beta-giardin (ß-giardin) have been widely used for genotyping. In addition, different genetic targets have been proposed as a valuable alternative to assess diversity and genetics of this microorganism. Thus, our objective was to evaluate new markers for the study of the diversity and intra-taxa genetic structure of G. intestinalis in silico and in DNA obtained from stool samples. We analysed nine constitutive genes in 80 complete genome sequences and in a group of 24 stool samples from Colombia. Allelic diversity was evaluated by locus and for the concatenated sequence of nine loci that could discriminate up to 53 alleles. Phylogenetic reconstructions allowed us to identify AI, AII and B assemblages. We found evidence of intra- and inter-assemblage recombination events. Population structure analysis showed genetic differentiation among the assemblages analysed.

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.

Molecular detection and genotyping of pathogenic protozoan parasites in raw and treated water samples from southwest Colombia.

BACKGROUND: Protozoan parasites such as Giardia duodenalis, Cryptosporidium spp., Cyclospora cayetanensis, Toxoplasma gondii and Entamoeba histolytica represent a great challenge to the systems producing water for human consumption because their cystic forms are persistent in the environment and resist to the disinfection methods conventionally used for their control. In this study, we investigated the presence of these protozoan pathogens in both raw and treated water samples used for the production of drinking water in Nariño Department, southwest Colombia. We collected 110 water samples (10 lof each sample) and analyzed them with real-time PCR (qPCR). qPCR-positive samples were genotyped with PCR and DNA sequencing. RESULTS: Giardia duodenalis was detected in 35/110 (31.8%) of the samples and Cryptosporidium spp. in 9/110 (8.2%) of the samples; no sample was positive for T. gondii, E. histolytica or C. cayetanensis. Giardia duodenalis was detected in samples of both raw water (Drinking Water Treatment Plants (DWTP): 47.83%;Drinking Water Rural Plants (DWRP): 18.42%) and water collected either after conventional physicochemical treatment (26.09%) or after disinfection by chlorine (50%), whereas Cryptosporidium spp. were only detected in raw waters (DWTP: 17.39%; DWRP: 13.16%). The two pathogens were detected in both types of treatment plants supplying water to urban areas and to rural zones. Analysis of gdh and tpi markers identified assemblages AI, AII and H of G. duodenalis, while analysis of the small subunit rRNA and gp60 markers of Cryptosporidium-positive samples identified C. parvum (Subtype IIcA5G3c), C. galli, C. molnari, Cryptosporidium sp. genotype II of bats and Cryptosporidium sp. genotype VIII of birds. CONCLUSIONS: The results obtained demonstrate the presence of protozoan parasites in the water of the study region, and the need to improve the surveillance systems for these pathogens and identify the corresponding sources of contamination.

Elimination of onchocerciasis from Colombia: first proof of concept of river blindness elimination in the world.

BACKGROUND: Onchocerciasis is a chronic parasitic infection originally endemic in 13 discrete regional foci distributed among six countries of Latin America (Brazil, Colombia, Ecuador, Guatemala, Mexico and Venezuela). In Colombia, this disease was discovered in 1965 in the Pacific Coast of the country. The National Onchocerciasis Elimination Program was established in 1993 with the aim of eliminating disease morbidity and infection transmission. In 2013, the World Health Organization (WHO) verified Colombia as free of onchocerciasis, becoming the first country in the world to reach such a goal. This report provides the empirical evidence of the elimination of Onchocerca volvulus transmission by Simulium exiguum (s.l.) after 12 years of 6-monthly mass drug administration of Mectizan® (ivermectin) to all the eligible residents living in this endemic area. METHODS: From 1996 onwards, a biannual community-based mass ivermectin administration programme was implemented, complemented by health education and community participation. In-depth parasitological, serological and entomological surveys were conducted periodically between 1998 and 2007 to evaluate the impact of ivermectin treatment according to the 2001 WHO guidelines. When the interruption of parasite transmission was demonstrated, the drug distribution ceased and a three-year post-treatment surveillance (PTS) period (2008-2010) was initiated. RESULTS: After 23 rounds of treatment, parasitological and ophthalmological assessments showed absence of microfilariae in skin and anterior chamber of the eyes. Serological tests proved lack of antibodies against O. volvulus in children under 10 years-old. A total of 10,500 S. exiguum flies tested by PCR had no L3 infection (infectivity rate = 0.0095%; 95% CI: 0.0029-0.049) during 2004, indicating interruption of parasite transmission. However, biannual ivermectin treatments continued until 2007 followed by a 3-year PTS period at the end of which 13,481 flies were analyzed and no infective flies were found (infectivity rate = 0%; 95% CI: 0.0-0.014). CONCLUSIONS: These results fulfilled the WHO criteria for onchocerciasis elimination. Consequently, in 2013 Colombia was verified as free of onchocerciasis, demonstrating that elimination of this neglected tropical disease is an achievable goal and paving the way for an elimination agenda to be followed by other endemic countries in Latin America and Africa.

[Profiles of intestinal polyparasitism in a community of the Colombian Amazon region].

INTRODUCTION: Intestinal parasite infections and polyparasitism are highly prevalent in the most vulnerable populations of developing countries due to environmental, biological and social determinants common in the transmission of parasites. Children between 1 and 15 years of age are the most affected population. OBJECTIVE: To describe the prevalence and profiles of intestinal polyparasitism in 1 to 15 year-old children from native communities in the Colombian Amazon region. MATERIALS AND METHODS: We used a non-probability sampling of 300 children between 1 and 15 years of age from several rural settlements and the main urban area of Puerto Nariño, Amazonas, Colombia, who participated voluntarily in the study. We obtained fecal samples that were analyzed by the direct method (0.85% saline solution-lugol) and the Kato-Katz technique. The most prevalent polyparasitism profiles were identified using Cohen's kappa coefficient with a 95% confidence interval. RESULTS: The prevalence of polyparasitism, defined as the presence of at least two intestinal pathogenic parasites, was 84% (95 % CI: 79.35-87.96). Polyparasitism by two or three of the following parasites: Ascaris lumbricoides, Trichuris trichiura, Blastocystis sp., Ancylostoma duodenale and Necator americanus from the Ancylostomatidae family, and Entamoeba histolytica, E. dispar and E. moshkovskii of the Entamoeba complex, had the highest prevalence. CONCLUSION: Several intestinal polyparasitism profiles were found, although in most cases fewer than six parasites were involved. Better prevalence estimations and identification of determinant factors will allow to priorize and direct resources to control these infections.

Perfiles de poliparasitismo intestinal en una comunidad de la Amazonia colombiana / Profiles of intestinal polyparasitism in a community of the Colombian Amazon region

Resumen Introducción. Las infecciones parasitarias y el poliparasitismo intestinal son muy prevalentes en las poblaciones más vulnerables de los países en desarrollo debido a la existencia de factores ambientales, biológicos y sociales determinantes en la transmisión de parásitos. La población entre uno y 15 años de edad es la más afectada por dicha situación. Objetivo. Describir las prevalencias y los perfiles del poliparasitismo intestinal en la población de uno a 15 años de edad de comunidades indígenas de la Amazonia colombiana. Materiales y métodos. Se hizo un muestreo no probabilístico de 300 niños entre uno y 15 años de edad, procedentes de varios asentamientos rurales y de la cabecera urbana de Puerto Nariño, Amazonas, quienes participaron voluntariamente en el estudio. Las muestras de materia fecal se analizaron mediante el método directo (solución salina al 0,85 % y lugol) y la técnica de Kato-Katz. Se determinaron los perfiles más prevalentes de poliparasitismo intestinal con el coeficiente kappa de Cohen y un intervalo de confianza de 95 %. Resultados. La prevalencia de poliparasitismo, definida como la presencia de, por lo menos, dos parásitos intestinales patógenos, fue de 84 % (IC95 %: 79,35-87,96). Los casos con presencia de dos o tres agentes incluyeron los parásitos Ascaris lumbricoides, Trichuris trichiura, Blastocystis sp., los ancilostomídeos Ancylostoma duodenale yNecator americanus, así como Entamoeba histolytica,E. dispar y E. moshkovskii del complejo Entamoeba, y fueron los de mayor prevalencia. Conclusión. Se encontraron varios perfiles de poliparasitismo intestinal, aunque la mayoría de los casos involucraron menos de seis especies. Una mejor estimación de la prevalencia y los factores determinantes del poliparasitismo intestinal permitirían priorizar y orientar los recursos para su control.

Abstract Introduction: Intestinal parasite infections and polyparasitism are highly prevalent in the most vulnerable populations of developing countries due to environmental, biological and social determinants common in the transmission of parasites. Children between 1 and 15 years of age are the most affected population. Objective: To describe the prevalence and profiles of intestinal polyparasitism in 1 to 15 year-old children from native communities in the Colombian Amazon region. Materials and methods: We used a non-probability sampling of 300 children between 1 and 15 years of age from several rural settlements and the main urban area of Puerto Nariño, Amazonas, Colombia, who participated voluntarily in the study. We obtained fecal samples that were analyzed by the direct method (0.85% saline solution-lugol) and the Kato-Katz technique. The most prevalent polyparasitism profiles were identified using Cohen's kappa coefficient with a 95% confidence interval. Results: The prevalence of polyparasitism, defined as the presence of at least two intestinal pathogenic parasites, was 84% (95 % CI: 79.35-87.96). Polyparasitism by two or three of the following parasites: Ascaris lumbricoides,Trichuris trichiura, Blastocystis sp., Ancylostoma duodenale and Necator americanus from the Ancylostomatidae family, and Entamoeba histolytica,E. dispar and E. moshkovskii of the Entamoeba complex, had the highest prevalence. Conclusion: Several intestinal polyparasitism profiles were found, although in most cases fewer than six parasites were involved. Better prevalence estimations and identification of determinant factors will allow to priorize and direct resources to control these infections.

Combination of five diagnostic tests to estimate the prevalence of hookworm infection among school-aged children from a rural area of colombia.

BACKGROUND: Public health programs for the control of soil-transmitted helminthiases require valid diagnostic tests for surveillance and parasitic control evaluation. However, there is currently no agreement about what test should be used as a gold standard for the diagnosis of hookworm infection. Still, in presence of concurrent data for multiple tests it is possible to use statistical models to estimate measures of test performance and prevalence. The aim of this study was to estimate the diagnostic accuracy of five parallel tests (direct microscopic examination, Kato-Katz, Harada-Mori, modified Ritchie-Frick, and culture in agar plate) to detect hookworm infections in a sample of school-aged children from a rural area in Colombia. METHODS AND RESULTS: We used both, a frequentist approach, and Bayesian latent class models to estimate the sensitivity and specificity of five tests for hookworm detection, and to estimate the prevalence of hookworm infection in absence of a Gold Standard. The Kato-Katz and agar plate methods had an overall agreement of 95% and kappa coefficient of 0.76. Different models estimated a sensitivity between 76% and 92% for the agar plate technique, and 52% to 87% for the Kato-Katz technique. The other tests had lower sensitivity. All tests had specificity between 95% and 98%. The prevalence estimated by the Kato-Katz and Agar plate methods for different subpopulations varied between 10% and 14%, and was consistent with the prevalence estimated from the combination of all tests. The Harada-Mori, Ritchie-Frick and direct examination techniques resulted in lower and disparate prevalence estimates. Bayesian approaches assuming imperfect specificity resulted in lower prevalence estimates than the frequentist approach.

Preservación de huevos de Necator americanus con acetato de sodio-ácido acético-formalina (SAF). Estudio de caso / Preservation of Necator americanus eggs with sodium acetate-acetic acid-formalin(SAF). Case study

Resumen Introducción. La técnica de Kato Katz es el método recomendado para realizar el recuento de huevos en materia fecal y determinar la intensidad parasitaria por helmintos. La calidad de los resultados se ve afectada por la tardanza en el procesamiento de la muestra, por lo que se requiere de preservantes que faciliten la lectura posterior a las 24 horas de recolección de la muestra. Objetivo. Demostrar validez de la solución con acetato de sodio-ácido acético-formalina (SAF) como preservante de huevos de Necator americanus por medio del estudio de un caso clinico. Materiales y métodos. Descripción de caso clínico con diagnóstico de parásitos intestinales por coprológico, intensidad de la infección por la técnica de Kato Katz, evaluación de la presencia de huevos N. americanus por la técnica de Kato Katz durante 10 meses en muestra preservada en SAF y determinación de especie de Uncinaria sp. por qPCR. Resultados. Los huevos de N. americanus se conservan de manera adecuada en morfología durante los primeros cinco meses. Conclusiones. A fin de ejecutar la técnica de Kato Katz, la fijación con SAF demostró ser adecuada para el mantenimiento de la intensidad parasitaria y morfología de los huevos de N. americanus en un período de hasta cinco meses.

Abstract Introduction: The Kato Katz technique is the recommended method for fecal egg counts and determining the intensity of parasitic helminths. The quality of the results is affected by the delay in the analysis of the sample. Therefore, fixatives are necessary to make the reading easier 24 hours after sample collection. Objective: To prove the usefulness of sodium acetate-acetic acid-formalin (SAF) solution as a fixative for Necator americanus eggs by means of a clinical case study. Materials and methods: Clinical case description and intestinal parasites diagnosis by direct stool microscopic analysis and parasite infection intensity by means of the Kato Katz technique. N. americanus eggs were detected by Kato Katz method for 10 months on a SAF-fixed stool sample. Identification of Uncinaria sp. by qPCR. Results: N. americanus eggs are adequately preserved in SAF during the first five months. Conclusions: SAF fixative is suitable for Kato Katz method and proved to be capable of maintaining parasite intensity and N. americanus egg morphology over a period of up to five months.

Geographic distribution of human Blastocystis subtypes in South America.

Blastocystis is a cosmopolitan enteric protist colonizing probably more than 1 billion people. This protozoan exhibits genetic diversity and is subdivided into subtypes (STs). The aim of this study was to determine the distribution of Blastocystis STs in symptomatic and asymptomatic human samples from different countries of South America. A total of 346 fecal samples were genotyped by SSU rDNA showing ST1 (28.3%), ST2 (22.2%), ST3 (36.7%), ST4 (2%), ST5 (2.3%), ST6 (2%), ST7 (2.3%), ST8 (0.6%), ST12 (0.9%) and a novel ST (2.7%). These findings update the epidemiology of Blastocystis in South America and expand our knowledge of the phylogeographic differences exhibited by this stramenopile.

Identification of Entamoeba moshkovskii in Treated Waste Water Used for Agriculture.

We conducted an observational study to determine the prevalence of Entamoeba spp., in samples collected in a waste water treatment plant that provides water for agricultural irrigation. Samples were collected weekly over a period of 10 weeks at representative contamination stages from within the treatment plant. Protozoan identification was performed via light microscopy and culture. PCR amplification of small subunit rRNA gene sequences of E. histolytica/dispar/moshkovskii was performed in culture positive samples. Light microscopy revealed the presence of Entamoeba spp., in 70% (14/20) of the raw waste water samples and in 80% (8/10) of the treated water samples. PCR amplification after culture at both 24 and 37°C revealed that 100% (29/29) of the raw waste water samples and 78.6% (11/14) of the treated waste water were positive for E. moshkovskii. We report the first isolation of E. moshkovskii in Colombia, confirmed by PCR. Recent reports of E. moshkovskii pathogenic potential suggest this finding could constitute a public health risk for people exposed to this water.

Molecular Epidemiology of Entamoeba: First Description of Entamoeba moshkovskii in a Rural Area from Central Colombia.

BACKGROUND: Entamoeba histolytica, E. dispar and E. moshkovskii are the most frequent species described in human infection where E. histolytica is the only true pathogen. The epidemiology of this infection is complex due to the absence of a routine exam that allows a correct discrimination of the Entamoeba species complex. Therefore, molecular methods appear as the unique epidemiological tool to accomplish the species discrimination. Herein, we conducted a cross-sectional study to determine the frequency of Entamoeba species infections in a group of asymptomatic individuals from a rural area in central Colombia. METHODOLOGY/PRINCIPAL FINDINGS: A total of 181 fecal samples from asymptomatic children under 16 years old from the hamlet La Vírgen, Cundinamarca (Colombia) that voluntarily accepted to participate in the study were collected. The fecal samples were examined by light microscopy and DNA-extracted, subsequently submitted to molecular discrimination of E. dispar/E. histolytica/E. moshkovskii infection based on a multiplex PCR assay targeting the 18S rRNA fragment. To confirm the species description, twenty samples were randomly submitted to DNA sequencing of the aforementioned fragment. By direct microscopic examination, frequency of the complex E. histolytica/E. dispar/E. moshkovskii was 18.8% (34/181). PCR showed a frequency of 49.1% (89/181), discriminated as 23.2% (42/181) that were positive for E. dispar, 25.4% (46/181) for E. moshkovskii and 0.55% (1/ 181) for E. histolytica. Also, mixed infections were detected between E. dispar and E. moshkovskii at 4.42% (8/181) of the samples. Molecular barcoding confirmed the diagnosis depicted by the multiplex PCR assay. CONCLUSIONS/SIGNIFICANCE: This is the first description of E. moshkovskii in Colombia and the second report in South-America to our knowledge. Our results suggest the need to unravel the true epidemiology of Entamoeba infections around the world, including the real pathogenic role that E. moshkovskii may have.

Molecular diagnosis and genotype analysis of Giardia duodenalis in asymptomatic children from a rural area in central Colombia.

Giardiasis is a parasitic infection that affects around 200 million people worldwide. This parasite presents a remarkable genetic variability observed in 8 genetic clusters named as 'assemblages' (A-H). These assemblages are host restricted and could be zoonotic where A and B infect humans and animals around the globe. The knowledge of the molecular epidemiology of human giardiasis in South-America is scarce and also the usefulness of PCR to detect this pathogen in fecal samples remains controversial. The aim of this study was to conduct a cross-sectional study to compare the molecular targets employed for the molecular diagnosis of Giardia DNA and to discriminate the parasite assemblages circulating in the studied population. We analyzed 181 fecal samples from Children at La Virgen, Cundinamarca, Colombia that were DNA-extracted and analyzed by SSU rDNA, tpi and gdh loci. We observed positivity by microscopy of 13% and by PCR around 76-80% depending on the molecular marker. Additionally, a lack of statistical concordance between microscopy and PCR was detected. Regarding the genetic assemblages, we detected assemblage A (3%), assemblage B (90%) and mixed infections assemblages A+B (7%). Hence, the sub-assemblages were typed as AI, AII, BIII and BIV across the population. This study represents a reliable attempt to understand the molecular epidemiology of giardiasis in Colombia and the use of PCR to detect cryptic infections. The epidemiological implications are herein discussed.

Comparison of hemocytes of V-instar nymphs of Rhodniusprolixus (Stål) and Rhodnius robustus (Larousse 1927), before and after molting / Comparación de los hemocitos de las ninfas de V instar de Rhodnius prolixus (Stål) and Rhodnius robustus, antes y después de la muda (Larousse 1927)

Background. The immune response of insects involves humoral factors and cellular elements known as hemocytes. There are different reports that explore the response of hemocytes to infections, but the effect that molting has on this response has not been explored so far. We hypothesized that there would be a change in the percentage of hemocytes as a response to the molting process. Objective. The aim of this work was to compare the hemogram (CBC), the formula, and the differential count of hemocytes in IV instar before molting and in V instar nymphs 24 hours after molting in two species: Rhodnius prolixus and Rhodnius robustus. Materials and methods. We assayed different staining methodologies including Giemsa, Alcian Blue pH 2.0, Alcian Blue pH 2.6, Gomori substrate, PAS (Schiff), Sudan Black and Papanicolau with positive controls for each one. In the Gomori staining, we observed lysosomes in the granulocytes and plasmatocytes, but the differentiation was better detected using Giemsa staining. Results. There were no statistically significant differences between the two species studied in plasmatocytes (p=0,053) or even in granulocytes (p=0,5). However, differences were significant in the prohemocytes (p=0,001) during the molting process in both Rhodnius prolixus and Rhodnius robustus. Conclusions. Significant differences in prohemocytes between nymphs of IV and V instar were detected. No significant differences in the amount of cells were observed between the two species and the two stages. These findings may be explained due to their role as precursor cell of prohemocytes.

Antecedentes. La respuesta inmune de los insectos involucra factores humorales y elementos celulares llamados hemocitos. Existen varios trabajos explorando la respuesta de los hemocitos frente a infecciones, pero no como respuesta al proceso de muda. Objetivo. Comparar el hemograma: la fórmula y el recuento diferencial de hemocitos de Rhodnius prolixus y Rhodnius robustus en ninfas de IV estadio antes de la muda y ninfas de V estadio 24 horas después de la muda. Materiales y métodos. Se ensayaron las coloraciones Giemsa, Alcian Blue pH 2.0, Alcian Blue pH 2.6, sustrato Gomori, PAS (Schiff), Sudán Negro, Papanicolau, con controles positivos para cada una. Con la coloración de Gomori se observaron lisosomas en los granulocitos y en plasmatocitos, pero la diferenciación se logró mejor con la coloración de Giemsa. Resultados. Los plasmatocitos fueron más abundantes en las ninfas de IV estadio de Rhodnius robustus y en las de V estadio de las dos especies, pero en las ninfas de V estadio de R. prolixus, el recuento de prohemocitos fue mayor; para los oenocitoides, esferulocitos y adipohemocitos el recuento no alcanzó al 1 %. En las dos especies no se encontraron diferencias estadísticamente significativas en plasmatocitos (p=0,05319), ni en granulocitos (p=0,5), entre las ninfas de IV y V estadio, pero fue significativa en prohemocitos (p=0,001). Conclusiones. Se detectaron diferencias significativas en prohemocitos entre las ninfas de IV y V estadio. No hubo un aumento significativo en el conteo de células en los diferentes estadios.

[Evaluation (assessment) of three tests for diagnosis of geohelmints in Colombia]. / Evaluación de tres pruebas para el diagnóstico de geohelmintos en Colombia.

INTRODUCTION: Soil-transmitted helminth infections are considered a public health problem in developing countries. The diagnostic tests, both for individual parient diagnosis as for population studies should be evaluated in terms of validity and reliability. OBJECTIVE: To compare the direct examination, the modified Ritchie-Frick method, a Kato-Katz designed by a Brazilian group and one designed by the WHO, for the diagnosis of soil-transmitted helminthes. MATERIALS AND METHODS: A diagnostic test reliability study was performed. The same stool sample was analyzed by the same observer using four diagnostic tests. 204 samples were obtained, 194 of those fulfilled the inclusion criteria and were analyzed. The observers did not know the participants' identity neither the other tests results. For the analysis the Kato-Katz (WHO) was considered as the gold standard. For the reliability assessment percent agreement, positive percent agreement, Kappa statistic, and intraclass correlation were performed. RESULTS: The Brazilian Kato-Katz showed a good performance with high sensitivity and specificity for T. trichiura and Hookworm with values of 0.97 and 0.96 respectively, and a high specificity with mild sensitivity for A. lumbricoides (0.95 and 0.79) meanwhile the direct examination and the Ritche-Frick method showed a performance between mild and poor. The differences were higher for hookworm and Trichiuris trichiura than for Ascaris lumbricoides. CONCLUSION: The Brazilian Kato Katz test could be implemented, but further studies are needed to correlate its operative capacity with its feasibility, availability and cost.

Entamoeba moshkovskii perspectives of a new agent to be considered in the diagnosis of amebiasis.

During the last decade Entamoeba moshkovskii has become relevant given its capacity to infect humans, especially when considering that it is morphologically indistinguishable from E. histolytica. For a long time, E. moshkovskii was considered as a free living amoeba, but in the last decade it has been demonstrated that E. moshkovskii can infect humans and can be found more frequently in regions where amebiasis shows high prevalence values, becoming a challenge to differentiate it from the E. histolytica/E. dispar complex. Recently there have been studies that raise the possibility that E. moshkovskii could be a pathogenic species, as there are reports in different countries that associated this infection with gastrointestinal symptoms even though others have described it as a non pathogenic species. For this reasons, both clinical and epidemiological studies are required.

Correlación entre la incidencia de malaria y la prevalencia de las geohelmintiasis en Colombia: enfoque ecológico / Correlation between malaria incidence and prevalence of soil-transmitted helminths in Colombia: An ecologic evaluation

Introducción. Los estudios recientes han sugerido una asociación entre las geohelmintiasis y la incidencia de malaria, sin embargo, la evidencia publicada es escasa y divergente. Desde 1977 no se han realizado nuevos estudios ecológicos para explorar esta asociación. Los estudios ecológicos podrían explorar dicha correlación en la población, midiendo su impacto potencial en salud pública. Objetivo. Explorar la asociación entre la prevalencia de geohelmintiasis y la incidencia de malaria desde el punto de vista ecológico, en Colombia. Materiales y métodos. Usando datos provenientes de la Encuesta Nacional de Salud, llevada a cabo en 1980, se estimaron coeficientes de correlación de Spearman a nivel departamental, entre las prevalencias de Ascaris lumbricoides, Trichuris trichiura y Uncinaria sp. con la incidencia de malaria para el mismo año suministrada por el Servicio de Erradicación de Malaria. A todos los datos, se les aplicó un método sólido de regresión con mínimos cuadrados ajustados.Resultados. La incidencia de malaria por Plasmodium falciparum y la prevalencia de A. lumbricoides tuvieron una correlación baja (R2=0,086). No obstante, dicha correlación se hizo más fuerte cuando se incluyó solamente el grupo de poblaciones con prevalencias de A. lumbricoides mayores de 30% (R2=0,916). Conclusión. Este trabajo evidenció una correlación ecológica en Colombia entre la incidencia de malaria y la prevalencia de geohelmintiasis. Esto podría justificar la existencia de una asociación entre estos dos grupos de parásitos o explicarse por la presencia de factores determinantes estructurales comunes a ambas enfermedades.

Introduction. Recent studies have suggested an association between the soil-transmitted helminth infections and malaria incidence. However, published evidence is still insufficient and diverging. Since 1977, new ecologic studies have not been carried out to explore this association. Ecologic studies could explore this correlation on a population level, assessing its potential importance on public health.Objectives. The aim of this evaluation is to explore the association between soil-transmitted helminths prevalence and malaria incidence, at an ecologic level in Colombia. Materials and methods. Using data from the National Health Survey, which was carried out in 1980 in Colombia, we calculated Spearman correlation coefficients between the prevalence of: Ascaris lumbricoides, Trichuris trichiura and hookworm, with the 1980 malaria incidence data of the same year provided from the Colombian Malaria National Eradication Service. A robust regression analysis with least trimmed squares was performed.Results. Falciparum malaria incidence and Ascaris lumbricoides prevalence had a low correlation (R2= 0.086) but this correlation was stronger into the clusters of towns with prevalence of Ascaris lumbricoides infection above 30% were only included (R2= 0.916). Conclusion. This work showed an ecologic correlation in Colombia between malaria incidence and soil-transmitted helminths prevalence. This could suggest that either there is an association between these two groups of parasites, or could be explained by the presence of common structural determinants for both diseases.

Correlation between malaria incidence and prevalence of soil-transmitted helminths in Colombia: an ecologic evaluation.

INTRODUCTION: Recent studies have suggested an association between the soil-transmitted helminth infections and malaria incidence. However, published evidence is still insufficient and diverging. Since 1977, new ecologic studies have not been carried out to explore this association. Ecologic studies could explore this correlation on a population level, assessing its potential importance on public health. OBJECTIVES: The aim of this evaluation is to explore the association between soil-transmitted helminths prevalence and malaria incidence, at an ecologic level in Colombia. MATERIALS AND METHODS: Using data from the National Health Survey, which was carried out in 1980 in Colombia, we calculated Spearman correlation coefficients between the prevalence of: Ascaris lumbricoides, Trichuris trichiura and hookworm, with the 1980 malaria incidence data of the same year provided from the Colombian Malaria National Eradication Service. A robust regression analysis with least trimmed squares was performed. RESULTS: Falciparum malaria incidence and Ascaris lumbricoides prevalence had a low correlation (R²= 0.086) but this correlation was stronger into the clusters of towns with prevalence of Ascaris lumbricoides infection above 30% were only included (R²= 0.916). CONCLUSION: This work showed an ecologic correlation in Colombia between malaria incidence and soil-transmitted helminths prevalence. This could suggest that either there is an association between these two groups of parasites, or could be explained by the presence of common structural determinants for both diseases.