A molecular and morphological study of Ascaris suum in a human-pig contact scenario in northeastern Brazil.

The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban communities in the state of Piauí, Brazil. Our cross-sectional survey included 100 fecal samples obtained from swine and 682 samples from humans. Fifteen pigs were necropsied. Human and porcine fecal samples were examined to identify Ascaris eggs. Parasites obtained in the swine necropsies were studied using scanning electron microscopy (SEM), and the mitochondrial gene encoding the cytochrome oxidase 1 (cox1) enzyme was partially amplified and sequenced for molecular taxonomy and phylogenetic analyses. The overall prevalence of Ascaris eggs in the swine fecal samples was 16/100 (16%). No Ascaris eggs were identified in the human fecal samples. SEM of six worms recovered from pigs demonstrated morphological characteristics of A. suum. Cox1 sequences were compatible with A. suum reference sequences. Original and reference (GenBank) nucleotide sequences were organized into clusters that did not segregate the parasites by host species or and region. The largest haplogroups were dominated by haplotypes H01, H02 and H31. In the communities studied, there was no epidemiological evidence of the zoonotic transmission of ascariasis at the human-swine interface.

A molecular and morphological study of Ascaris suum in a human-pig contact scenario in northeastern Brazil / Estudo molecular e morfológico de Ascaris suum em um cenário de contato entre suínos e humanos no nordeste brasileiro

The aim of the present study was to assess morphologic and genetic data on ascariasis in swine (Sus scrofa domesticus) and humans in low-resource rural and periurban communities in the state of Piauí, Brazil. Our cross-sectional survey included 100 fecal samples obtained from swine and 682 samples from humans. Fifteen pigs were necropsied. Human and porcine fecal samples were examined to identify Ascaris eggs. Parasites obtained in the swine necropsies were studied using scanning electron microscopy (SEM), and the mitochondrial gene encoding the cytochrome oxidase 1 (cox1) enzyme was partially amplified and sequenced for molecular taxonomy and phylogenetic analyses. The overall prevalence of Ascaris eggs in the swine fecal samples was 16/100 (16%). No Ascaris eggs were identified in the human fecal samples. SEM of six worms recovered from pigs demonstrated morphological characteristics of A. suum. Cox1 sequences were compatible with A. suum reference sequences. Original and reference (GenBank) nucleotide sequences were organized into clusters that did not segregate the parasites by host species or and region. The largest haplogroups were dominated by haplotypes H01, H02 and H31. In the communities studied, there was no epidemiological evidence of the zoonotic transmission of ascariasis at the human-swine interface.(AU)

O presente estudo teve como objetivo acessar dados morfológicos e genéticos sobre a ascaridíase em suínos (Sus scrofa domesticus) e humanos, em comunidades rurais e periurbanas no estado do Piauí. O estudo transversal incluiu 100 amostras fecais de suínos e 682 amostras obtidas de humanos. Quinze suínos foram necropsiados. Amostras fecais suínas e humanas foram examinadas para detecção de ovos de Ascaris. Os parasitas adultos, obtidos nas necropsias, foram estudados através de microscopia eletrônica de varredura (MEV), e o gene mitocondrial codificante da enzima citocromo oxidase 1 (cox1) foi parcialmente amplificado e sequenciado para análises filogenéticas e de taxonomia molecular. A prevalência de Ascaris em amostras fecais de suínos foi 16/100 (16%), não sendo identificado nenhum caso de infecção por este parasita em humanos. A análise por MEV de parasitas recuperados de suínos demonstrou características morfológicas de Ascaris suum. As sequências nucleotídicas de cox1 foram compatíveis com A. suum. As sequências originais e de referência (obtidas no GeneBank) foram organizadas em clusters que não segregaram os parasitas por hospedeiro ou região geográfica. Os maiores haplogrupos foram dominados pelos haplótipos H01, H02 e H31. Nas comunidades estudadas, não foi evidenciada transmissão zoonótica de A. suum na interface suíno-humana.(AU)

Cytochrome c oxidase subunit 1 gene reveals species composition and phylogenetic relationships of Oesophagostomum spp. infecting pigs in northeastern Brazil.

Helminths of the genus Oesophagostomum cause enteric diseases and affect domestic animals such as pigs. The aim of this study was to explore the species composition and genetic diversity of Oesophagostomum spp. infecting pigs in close contact with humans in the state of Piauí, Brazil. Eighty-seven fecal samples were collected for parasitological tests and molecular analysis. Through microscopy, the overall positivity rate for strongyliform eggs was 81.6% among the pigs studied. Forty-two strongyliform egg samples were subjected to PCR and six cox1 sequences (637 bp) were identified for the genus Oesophagostomum. The sequences were identified as Oesophagostomum dentatum, O. quadrispinulatum and O. columbianum. In the phylogenetic tree and haplotype network, 89 sequences were separated into seven clusters, which also included reference sequences from GenBank. Oesophagostomum dentatum and O. quadrispinulatum were seen to be closely related species and formed a monophyletic group related to O. aculeatum. Oesophagostomum columbianum showed similarity with sequences from parasites infecting small ruminants and the clade was positioned closer to O. bifurcum. High interspecific diversity was found and intraspecific diversity varied according to the species. This was the first study to characterize Oesophagostomum DNA sequences obtained from pigs in Brazil.

Epidemiology, species composition and genetic diversity of tetra- and octonucleated Entamoeba spp. in different Brazilian biomes.

BACKGROUND: Entamoeba species harbored by humans have different degrees of pathogenicity. The present study explores the intra- and interspecific diversity, phylogenetic relationships, prevalence and distribution of tetra- and octonucleated cyst-producing Entamoeba in different Brazilian regions. METHODS: Cross-sectional studies were performed to collect fecal samples (n = 1728) and sociodemographic data in communities located in four Brazilian biomes: Atlantic Forest, Caatinga, Cerrado, and Amazon. Fecal samples were subjected to molecular analysis by partial small subunit ribosomal DNA sequencing (SSU rDNA) and phylogenetic analysis. RESULTS: Light microscopy analysis revealed that tetranucleated cysts were found in all the studied biomes. The highest positivity rates were observed in the age group 6-10 years (23.21%). For octonucleated cysts, positivity rates ranged from 1 to 55.1%. Sixty SSU rDNA Entamoeba sequences were obtained, and four different species were identified: the octonucleated E. coli, and the tetranucleated E. histolytica, E. dispar, and E. hartmanni. Novel haplotypes (n = 32) were characterized; however, new ribosomal lineages were not identified. The Entamoeba coli ST1 subtype predominated in Atlantic Forest and Caatinga, and the ST2 subtype was predominant in the Amazon biome. E. histolytica was detected only in the Amazon biome. In phylogenetic trees, sequences were grouped in two groups, the first containing uni- and tetranucleated and the second containing uni- and octonucleated cyst-producing Entamoeba species. Molecular diversity indexes revealed a high interspecific diversity for tetra- and octonucleated Entamoeba spp. (H ± SD = 0.9625 ± 0.0126). The intraspecific diversity varied according to species or subtype: E. dispar and E. histolytica showed lower diversity than E. coli subtypes ST1 and ST2 and E. hartmanni. CONCLUSIONS: Tetra- and octonucleated cyst-producing Entamoeba are endemic in the studied communities; E. histolytica was found in a low proportion and only in the Amazon biome. With regard to E. coli, subtype ST2 was predominant in the Amazon biome. The molecular epidemiology of Entamoeba spp. is a field to be further explored and provides information with important implications for public health.

Socioenvironmental Factors Influencing Distribution and Intensity of Soil-Transmitted Helminthiasis in the Brazilian Amazon: Challenges for the 2030 Agenda.

Soil-transmitted helminthiasis (STHs) are poverty-related diseases with high prevalence rates in developing countries. The present study aims to describe the epidemiological scenario of STHs in an urban population in the Brazilian Amazon. A cross-sectional survey (n = 349 children aged 1-15 years) was carried out to obtain faecal samples and sociodemographic and sanitation data. Among the children, 143 (41%) were positive for at least one STH. Prevalence rates of infections by A. lumbricoides, T. trichiura, and hookworms were 24.4%, 42.6%, and 9%, respectively. A logistic regression multivariate model showed that infection with A. lumbricoides is significantly more frequent in children aged 11-15 years (odds ratio [OR] = 2.38; 95% confidence interval [CI] = 1.15-4.94; p=0.018) and the presence of latrines inside houses is a protection factor against ascariasis (OR = 0.38; 95% CI = 0.17-0.85; p=0.019). Positivity for T. trichiura is higher in the 5-10 (OR = 3.31; 95% IC = 1.85-5.89; p=0.001) and 11-15 age groups (OR = 3.16; 95% IC = 1.66-6.00; p=0.001), in children living in poor families (OR = 1.78; 95% IC = 1.01-3.14; p=0.045) and practicing open evacuation (OR = 2.07; 95% IC = 1.07-3.99; p=0.029). Hookworm infection is more frequent in children aged 11-15 years (OR = 6.70; 95% IC = 1.91-23.43; p=0.002), males (OR = 6.35; 95% IC = 2.00-20.14; p=0.002), and those living in stilt houses (OR = 3.52; 95% IC = 1.22-10.12; p=0.019). The use of albendazole in the last six months was a protection factor against hookworm infection (OR = 0.31; 95% IC = 0.10-0.96; p=0.042). The proportion of mild, moderate, and severe infections was 55.2%, 37.8%, and 7%, respectively, for A. lumbricoides, 72.4%, 24.3%, and 3.3% for T. trichiura, and 93.8%, 3.1%, and 3.1% for hookworms. Significantly higher worm burdens in T. trichiura and hookworm infections were associated with practicing open defecation and living in stilt houses. The data points to the need to improve sanitation infrastructure in Amazonian cities with similar sociodemographic and environmental characteristics.

Mitochondrial DNA reveals species composition and phylogenetic relationships of hookworms in northeastern Brazil.

Hookworm infection persists focally in rural communities in Brazil. In this study, we analyze the mitochondrial nucleotide sequences obtained from hookworms infecting humans in order to characterize species composition and assess their genetic diversity and phylogenetic relationships. Field expeditions and cross-sectional surveys were carried out in three Brazilian municipalities from 2013 to 2017: Nossa Senhora de Nazaré (n = 605) and Teresina (n = 297), in the state of Piauí, and Russas (n = 213) in the State of Ceará. Parasitological methods were used to evaluate fecal samples. Hookworm-positive samples had a partial mtDNA cox1 amplified and sequenced. Maximum-likelihood and Bayesian analysis demonstrated two strongly-supported clades, including Group A, corresponding to Necator americanus, and Groups B and C, corresponding to Necator sp. Group A was divided into three main clusters: A1 grouped with Asian sequences, A2 grouped with African sequences, and A3 had only Asian sequences. Group B was closely related to Necator sp., showing a sequence similarity of 98%-99% with African samples circulating zoonotically among humans and non-human primates. Twenty three N. americanus haplotypes were identified. N. americanus Median-Joining network revealed three distinct groups, designated again as A1, A2, and A3. Group A1 presented a star-like shape, with one dominant haplotype. The molecular dating suggested that the two clades dividing N. americanus and Necator sp. began to diverge during the middle Pleistocene. The most recent common ancestor among N. americanus groups was dated to the late Pleistocene. Hookworms circulating in the studied communities are structured in well-defined subpopulations presenting both Asian and African genetic backgrounds. This reveals a double origin for hookworms in northeastern Brazil and opens up new possibilities in phylogeographic, evolutionary, and molecular epidemiological studies in regions where hookworms persists focally, despite control efforts. The presence of potentially zoonotic species and the specific identification of Necator sp. should be further investigated.

Focal persistence of soil-transmitted helminthiases in impoverished areas in the State of Piaui, Northeastern Brazil.

This study aims to describe the prevalence, distribution, and factors associated with soil-transmitted helminthiases (STHs) in rural localities in Piaui, Brazil. Two cross-sectional surveys (n=605 subjects; 172 families) were carried out in order to obtain socio-demographic, anthropometric, spatial and parasitological data. Parasites were evaluated using Kato-Katz and centrifugal sedimentation techniques. Eggs were measured to assess infection with zoonotic Strongylida parasites. Kernel maps were constructed with Q-GIS. The prevalence of hookworm infection was 12.4% (75/605). Other helminthes found were Trichuris trichiura (n=1; 0.2%) and Hymenolepis nana (n=1; 0.2%). The hookworm positivity rate was significantly lower among subjects who had used albendazole when compared with individuals who had not used anthelmintics or had used antiprotozoal drugs in the last 6 months (8/134 [6.0%] vs. 59/415 [14.2%]; p=0.009). A total of 39/172 (22.7%) families had at least one infected member. The association between the number of dwellers and hookworm positivity in the family was present in a logistic regression multivariate model. Assessment of worm burdens showed 92.2% light, 6.2% moderate, and 1.6% heavy infections. Hookworm eggs (n=34) measured 57.2 - 75.4 µm in length and 36.4 - 44.2 µm in width (mean ± SD = 65.86 ± 4.66 µm L and 40.05 ± 1.99 µm W), commensurate with human hookworms. Hotspots suggest that transmission has a focal pattern. STHs persist in impoverished rural areas in Northeastern Brazil where currently available control strategies (mass drug administration) apparently do not allow the elimination of the infection.

Genetic diversity of Giardia duodenalis circulating in three Brazilian biomes.

Giardia duodenalis has a wide genetic variety, and its characterization helps in the understanding of its transmission dynamics and in the development control strategies. This study aimed to assess the genetic diversity of G. duodenalis obtained in different Brazilian biomes and estimate their phylogenetic relationships. Three surveys including 944 participants were carried out in the municipalities of Russas (RSS, Caatinga semiarid biome), Santa Isabel do Rio Negro (SIRN, Amazon rainforest biome) and Nossa Senhora de Nazaré (NSN, Cerrado-Caatinga transition biome). G. duodenalis-positive fecal samples were submitted to amplification of gene fragments encoding ß-giardin (ßG, N = 71), glutamate dehydrogenase (GDH, N = 42), and triosephosphate isomerase (TPI, N = 27). Overall detection rates of assemblage A in G. duodenalis-positive samples through ßG, GDH and TPI were 22/71 (31%), 13/42 (31%), and 13/27 (48.1%), respectively. Concerning assemblage B, rates with distinct genetic markers were 49/71 (69%), 29/42 (69%), and 14/27 (51.9%), respectively. In the Amazon, assemblage B was more prevalent (77.8%, 71.8% and 65% through ßG, GDH and TPI, respectively), while in the Cerrado biome assemblage A predominated (50%, 66.6%, and 85.7%, through ßG, GDH and TPI, respectively). In Caatinga biome assemblage A also predominated (71.4%, through ßG). Thirty new sub-assemblages are described for assemblage B (24 ßG and six TPI), as well as three new sub-assemblages are described for assemblage A (one GDH and 2 TPI). Higher genetic diversity of assemblage B in the Amazon may be related to demographic concentration leading to a more complex transmission network within a poorer sanitation background. The high genetic divergence between assemblages A and B (5.5-6.3%) support the proposal of taxon separation in distinct species.

Giardia intestinalis infection associated with malnutrition in children living in northeastern Brazil.

INTRODUCTION: The present study aimed to determine the prevalence and factors associated with Giardia intestinalis infection, verifying its impact on the nutritional status of children in northeastern Brazil. METHODOLOGY: A cross-sectional study was conducted to obtain parasitological, sociodemographic, and anthropometric data in two municipalities in the states of Piauí and Ceará, northeastern Brazil. RESULTS: Prevalence of giardiasis was 55/511 (10.8%). G. intestinalis was more frequent in people living in poverty (30/209 [14.4%], p = 0.041), performing open evacuation (26/173 [15%], p = 0.034), and drinking rainwater stored in cisterns (9/56 [16.1%], p = 0.005). The proportion of stunting and being underweight in children infected with G. intestinalis was significantly higher than that in uninfected children (5/23 [21.7%] vs. 10/179 [5.6%], p = 0.017, OR = 4.69, 95% confidence interval [CI] = 1.44-15.25 and 5/23 [21.7%] vs. 13/179 [7.3%], p = 0.038, OR = 3.54, 95% CI = 1.13-11.09, respectively). Infection with G. intestinalis remained significantly associated with stunting and being underweight after adjustment for poverty, municipality, sex, and age in a logistic regression multivariate model. CONCLUSIONS: In rural areas in northeastern Brazil, giardiasis has acquired great public health importance in the soil-transmitted helminths control era, impacting the nutritional status of children and requiring new approaches to diagnosis and treatment and translational research that could generate applicable solutions at the community level.

Frequency and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, and Entamoeba hartmanni in the context of water scarcity in northeastern Brazil.

This study aimed to estimate the frequency, associated factors, and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, andEntamoeba hartmanni infections. We performed a survey (n = 213 subjects) to obtain parasitological, sanitation, and sociodemographic data. Faecal samples were processed through flotation and centrifugation methods.E. histolytica, E. dispar, E. moshkovskii, and E. hartmanni were identified by nested-polymerase chain reaction (PCR). The overall prevalence of infection was 22/213 (10.3%). The infection rate among subjects who drink rainwater collected from roofs in tanks was higher than the rate in subjects who drink desalinated water pumped from wells; similarly, the infection rate among subjects who practice open defecation was significantly higher than that of subjects with latrines. Out of the 22 samples positive for morphologically indistinguishableEntamoeba species, the differentiation by PCR was successful for 21. The species distribution was as follows: 57.1% to E. dispar, 23.8% to E. histolytica, 14.3% toE. histolytica and E. dispar, and 4.8% E. dispar and E. hartmanni. These data suggest a high prevalence of asymptomatic infection by the group of morphologically indistinguishable Entamoeba histolytica/dispar/moshkovskiicomplex and E. hartmanni species. In this context of water scarcity, the sanitary and socioenvironmental characteristics of the region appear to favour transmission.

Frequency and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, and Entamoeba hartmanni in the context of water scarcity in northeastern Brazil

This study aimed to estimate the frequency, associated factors, and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, andEntamoeba hartmanni infections. We performed a survey (n = 213 subjects) to obtain parasitological, sanitation, and sociodemographic data. Faecal samples were processed through flotation and centrifugation methods.E. histolytica, E. dispar, E. moshkovskii, and E. hartmanni were identified by nested-polymerase chain reaction (PCR). The overall prevalence of infection was 22/213 (10.3%). The infection rate among subjects who drink rainwater collected from roofs in tanks was higher than the rate in subjects who drink desalinated water pumped from wells; similarly, the infection rate among subjects who practice open defecation was significantly higher than that of subjects with latrines. Out of the 22 samples positive for morphologically indistinguishableEntamoeba species, the differentiation by PCR was successful for 21. The species distribution was as follows: 57.1% to E. dispar, 23.8% to E. histolytica, 14.3% toE. histolytica and E. dispar, and 4.8% E. dispar and E. hartmanni. These data suggest a high prevalence of asymptomatic infection by the group of morphologically indistinguishable Entamoeba histolytica/dispar/moshkovskiicomplex and E. hartmanni species. In this context of water scarcity, the sanitary and socioenvironmental characteristics of the region appear to favour transmission.

Parasitismo intestinal e fatores associados em quatro localidades rurais no semiárido nordestino brasileiro / Parasitism intestinal and associated factors in four locations in rural northeastern semiarid

As parasitoses intestinais são altamente prevalentes nas populações humanas, sendo endêmicas em países em desenvolvimento e representam importante problema de Saúde Pública. Diversos são os fatores associados àtransmissão destes parasitas, entre eles o saneamento precário ou ausente. Os objetivos deste estudo foram: i) estimar a prevalência e fatores associados às diferentes parasitoses intestinais em localidades rurais do estado do Ceará, ii) avaliar o estado nutricional das crianças da região, correlacionando-o com a infecção pelos diferentes parasitas intestinais e iii) realizar a caracterização molecular de amebas do complexo Entamoeba histolytica/ E.dispar/ E. moshkovskii e E. hartmanni identificadas nas amostras fecais. Trata-se de um estudo transversal, incluindo 326 moradores de quatro localidades rurais do município de Russas, Estado do Ceará. Os exames de fezes foram realizados por meio das técnicas de Lutz, Ritchie modificado por Young et al., Kato-Katz e BaermannMoraes.Dados sócio-demográficos e antropométricos foram obtidos em visitas domiciliares. A prevalência dos parasitas foi: 0,3 por cento para Strongyloides stercoralis, 1,4 por cento para Rodentolepis nana, 3,7 por cento para ancilostomídeos, 10,2 por cento para amebas do complexo Entamoeba histolytica/ E.dispar/ E. moshkovskii e E. hartmanni e 14 por cento para Giardia duodenalis; 39 por cento dos indivíduos estavam poliparasitados. Na ausência de programas de transferência de renda, 12,2 por cento das pessoas incluídas neste estudo apresentaram renda per capita familiar mensal inferior a R$ 70,00, podendo ser classificadas no extrato de pobreza extrema. As bolsas do governo beneficiavam 83,1 por cento dos participantes, sendo 46 por cento pelo programa Bolsa Família...

Observou-se que 65,3 por cento dos indivíduos faziam uso deágua de poço dessalinizada; 24,8 por cento utilizavam água armazenada da chuva coletada dos telhados via calhas ecisternas para o consumo e 7,3 por cento utilizavam água fornecida por caminhão-pipa; 21,7 por cento das pessoas evacuavam acéu aberto. A desnutrição crônica foi observada em 5/79 crianças (6,3 por cento) e o baixo peso em 4/76 crianças (5,3 por cento). Não houve associação entre a infecção por ancilostomídeos e G. duodenalis e a fonte de água para beber. Háuma maior taxa de positividade de infecção para Entamoeba histolytica/ E. dispar/ E. moshkovskii/ E. hartmannientre os indivíduos que defecam a céu aberto do que entre aqueles que defecam em banheiro (p = 0,032). Com relação à amebíase, a taxa de positividade foi maior entre os indivíduos que defecam a céu aberto (19,1 por cento) em comparação com os que defecam no banheiro (7,8 por cento), p=0,032. As taxas de infecção por G. duodenalis, Entamoeba histolytica/E.dispar/E. moshkovskii e E. hartmanni e ancilostomídeos tendem a ser mais baixas no estrato superior de renda. Não houve correlação entre a renda per capita familiar mensal e o estado nutricional das crianças para o parâmetro altura-idade (R = 0,148 e p = 0,194) e o parâmetro peso-idade (R = 0,080 e p = 0,495). Não houve diferença entre as médias do escore-Z do parâmetro antropométrico altura-idade das crianças infectadas por G. duodenalis e as não infectadas. Conclui-se que a ancilostomíase, a amebíase e a giardíase são as infecções intestinais mais prevalentes na região, havendo associação entre o parasitismo e o local de defecação...