Deteção de enteroparasitas e caracterização genética de Cryptosporidium spp. e Giardia duodenalis em crianças até aos 14 anos, com diarreia, em Moçambique

As infeções parasitárias causadas por helmintas intestinais, estão entre as mais prevalentes infeções lentas em humanos no mundo, principalmente nos países em desenvolvimento, incluindo Moçambique e, com as infeções causadas por protozoários intestinais, contribuem significativamente para grande carga de doenças gastrointestinais nestes países, sobretudo nas crianças. Giardia duodenalis e Cryptosporidium spp. são dois organismos protozoários que infetam os humanos e diversas espécies animais e são conhecidos por apresentar um elevado potencial em causar diarreia. Existem em Moçambique alguns estudos sobre a epidemiologia das infeções provocadas por parasitas intestinais, contudo, esta informação é muito fragmentada e escassa sobretudo no que diz respeito à epidemiologia molecular de G. duodenalis e Cryptosporidium spp. Este estudo teve como objetivo geral, determinar a frequência e os fatores relacionados com a infeção por parasitas intestinais (IPI) e caracterizar geneticamente G. duodenalis e Cryptosporidium spp. circulantes em crianças até aos 14 anos de idade com doença diarreica, nas regiões sul, centro e norte de Moçambique. Para o estudo foram analisadas 1424 amostras de fezes diarreicas (do projeto ViNaDia, Moçambique) recrutadas em crianças até 14 anos em seis hospitais públicos das regiões sul, centro e norte de Moçambique. Uma única amostra de fezes foi colhida por criança e examinada por Microscopia Óptica (M.O.) usando a concentração de formol-éter e a técnica de coloração de Ziehl-Neelsen modificada para identificar parasitas intestinais, nomeadamente coccídeos e Cryptosporidium. As características sociodemográficas foram obtidas por meio de questionários. Para caracterização molecular foram utilisadas amostras positivas por M.O. assim como por imunoensaio enzimático (ELISA) (de outro subestudo do projeto ViNaDia). A caracterização genética foi realizada por nested-PCR com alvos nos genes β-giardina de G. duodenlais e ssu rRNA e gp60 de Cryptosporidium spp. Análises uni e bivariadas e regressão logística binária foram realizadas para descrever a população e determinar os fatores de risco. Níveis de significância de 5% foram considerados significativos. Pelo menos uma IPI foi detetada em 19,2% (273/1424) das crianças. Cryptosporidium spp. foi o parasita mais comum (8,1%; 115/1424). Poliparasitismo foi observado em 26,0% (71/273) das crianças estudadas, sendo a coinfeção Ascaris lumbricoides e Trichuris trichiura (26,8%; 19/71) a mais comum. A idade e província foram relacionadas com a presença de IPI (p-valor < 0,05). A maior ocorrência de IPI foi observada na estação das chuvas (outubro a março) com 23,1% (191/828) em relação aos períodos secos (abril a setembro) com 13,6% (80/588) (p-valor < 0,001 ). O subtipo IaA9G3 de C. hominis foi o dominante nas crianças infetadas por Cryptosporidium e o subgenótipo AII de G. duodenalis foi o mais observado (56,6%; 13/23) na população pediátrica moçambicana. Cryptosporidium spp. e a combinação A. lumbricoides/T. trichiura foram os principais parasitas intestinais observados em crianças hospitalizadas com diarreia em Moçambique. Os dados de genotipagem sugerem que as principais vias de transmissão de G. duodenalis e Cryptosporidium em Moçambique são antroponóticas.Parasitic infections caused by intestinal helminths are among the most prevalent sluggish infections in humans in the world, mainly in developing countries, including Mozambique, and with infections caused by intestinal protozoa, they contribute significantly to large burdens of gastrointestinal diseases in these countries, especially in children. Giardia duodenalis and Cryptosporidium spp. are two protozoan organisms that infect humans and several animal species and are known to have a high potential to cause diarrhea. There are some studies in Mozambique on the epidemiology of diseases caused by intestinal parasites; however, this information is very fragmented and scarce regarding the molecular epidemiology of G. duodenalis and Cryptosporidium spp. This study aimed to determine the frequency and factors related to intestinal parasites infection and to characterize genetically G. duodenalis and Cryptosporidium spp. circulating in children up to 14 years of age with diarrheal disease in southern, central and northern Mozambique. For this purpose, 1424 diarrheal stool samples (included in the ViNaDia project, Mozambique) were collected from children up to 14 years old in six public hospitals in the southern, central and northern regions of Mozambique. A single stool sample was collected per child and examined by light microscopy using formalin-ether concentration and modified Ziehl-Neelsen techniques to identify intestinal parasites, namely coccidia and Cryptosporidium. Sociodemographic characteristics were obtained through questionnaires. For molecular characterization positive samples from light microscopy as well as by ELISA (from another ViNaDia sub study) were used. Genetic characterization was performed by nested-PCR targeting β-giardin gene for G. duodenlais and SSUrRNA and gp60 genes for Cryptosporidium spp. Univariate and bivariate analysis and binary logistic regression were performed to describe the population and determine risk factors. Significance levels of 5% were considered significant. A single IPI was detected in 19.2% (273/1424) of children. Cryptosporidium spp. was the most common parasite (8.1%; 115/1424). Polyparasitism was observed in 26.0% (71/273) of the children, with Ascaris lumbricoides and Trichuris trichiura (26.8%; 19/71) being the most common co-infection. Age and province were related to IPI (p-value < 0.05). The highest occurrence of IPI was observed in the wet season (October to March) with 23.1% (191/828) compared to the dry season (April to September) with 13.6% (80/588) (p-value < 0.001). The IbA9G3 subtype of C. hominis was the most dominant in children infected with Cryptosporidium and sub assemblage AII 56.6% (13/23) more frequently observed in children infected with G. duodenalis in the Mozambican pediatric population. Cryptosporidium spp. and the combination A. lumbricoides/T. trichiura were the main intestinal parasites observed in children hospitalized with diarrhea in Mozambique. Genotyping data suggest that the main routes of transmission of G. duodenalis and Cryptosporidium in Mozambique are anthroponotic

Intestinal parasites in children up to 14 years old hospitalized with diarrhea in Mozambique, 2014–2019

© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).Diarrhea remains a public health problem in Mozambique, even with control strategies being implemented. This analysis aimed to determine the proportion and factors associated with intestinal parasitic infection (IPI) in children up to 14 years old with diarrheal disease, in the southern, central and northern regions of Mozambique. A single diarrheal sample of 1424 children was collected in hospitals and examined using the formol-ether concentration and modified Ziehl-Neelsen techniques to identify intestinal parasites using optical microscopy. Sociodemographic characteristics were obtained by questionnaires. Descriptive statistics and cross-tabulation were performed, and p-values <0.05 were considered statistically significant. A single IPI was detected in 19.2% (273/1424) of the children. Cryptosporidium spp. was the most common parasite (8.1%; 115/1424). Polyparasitism was seen in 26.0% (71/273), with the co-infection of Ascaris lumbricoides and Trichuris trichiura (26.8%; 19/71) being the most common. Age and province were related to IPI (p-value < 0.05). The highest occurrence of IPI was observed in the wet period (October to March), with 21.9% (140/640), compared to the dry period (April to September), with 16.9% (131/776) (p-value = 0.017). Cryptosporidium spp. and the combination of A. lumbricoides/T. trichiura were the main intestinal parasites observed in children hospitalized with diarrhea in Mozambique.This work was supported by funds from the European Foundation Initiative for African Research into Neglected Tropical Diseases (EFINTD, grant number 98539), the World Health Organization, Deutsche Forschungsgemeinschaft (DFG, grant number JO369/5-1) and The Global Vaccine Alliance Initiative through Health System Strengthening. O.N., PhD, is supported by Camões—Instituto da Cooperação e da Língua.info:eu-repo/semantics/publishedVersio