Toxoplasma gondii and Neospora caninum antibody seroprevalence and risk factors among dogs treated at Public Veterinary Hospitals in São Paulo, Brazil / Soroprevalência de anticorpos anti-Toxoplasma gondii e Neospora caninum em cães atendidos em Hospitais Públicos Veterinários em São Paulo, Brasil

Abstract Dogs can be infected by Toxoplasma gondii and Neospora caninum, for which they function, respectively, as intermediate, and definitive hosts. In the present study seroprevalence against T. gondii and N. caninum antibodies, were determined by indirect fluorescent antibody test (cut off of 16 and 50, respectively), in dogs that were treated at public veterinary hospitals in the metropolitan region of São Paulo and risk factors were identified. Out of the 1,194 samples 125 (10.5%; 95% CI: 8.8-12.3%) were positive for T. gondii and 9 (0.75%, 95% CI: 0.34-1.4%) for N. caninum. For T. gondii, statistical differences were observed between the proportions of positive dogs and different zones of the municipality (p = 0.025), and age (p = 0.02), higher among older dogs. The keepers were invited to answer an epidemiological questionnaire to analyze risk factors, and 471 (39.4%) agreed to be interviewed, and among their dogs 65 (13.8%) were T. gondii seropositive. Age group above 8 years (OR = 3.63; 95% CI: 1.08-12.23) was a risk factor and having a defined breed (OR = 0.49; 95% CI: 0.25-0.96) was a protective factor for T. gondii infection. Because of the low number of dogs positive for N. caninum, risk factors for this coccidium were not determined.

Resumo Cães podem ser infectados por Toxoplasma gondii e Neospora caninum, os quais funcionam, respectivamente, como hospedeiros intermediários e definitivos. Neste estudo, a soroprevalência contra anticorpos anti-T. gondii e N. caninum foi determinada pelo teste de imunofluorescência indireta (ponto de corte de 16 e 50, respectivamente), em cães atendidos em hospitais públicos veterinários da região metropolitana de São Paulo e fatores de risco foram identificados. Das 1.194 amostras, 125 (10,5%; IC 95%: 8,8-12,3%) foram positivas para T. gondii e 9 (0,75%, IC 95%: 0,34-1,4%) para N. caninum. Para T. gondii foram observadas diferenças entre as proporções de cães positivos e diferentes zonas do município (p = 0,025) e idade (p = 0,02), prevalência maior entre os mais velhos. Os tutores foram convidados a responder um questionário epidemiológico para análise de fatores de risco, e 471 (39,4%) concordaram em ser entrevistados, destes 65 cães (13,8%) eram soropositivos para T. gondii. Faixa etária acima de 8 anos (OR = 3,63; IC 95%: 1,08-12,23) foi fator de risco e raça definida (OR = 0,49; IC 95%: 0,25-0,96) foi fator de proteção para a infecção por T. gondii. Devido ao baixo número de positivos para anticorpos anti-N. caninum, fatores de risco para este coccídio não foram determinados.

Occurrence of gastrointestinal parasites in dogs in a rural area of Santa Catarina, Brazil / Ocorrência de parasitas gastrintestinais em cães de área rural de Santa Catarina, Brasil

Abstract We investigated the occurrence of gastrointestinal parasites in dogs in rural areas of the municipality of Painel, Santa Catarina, Brazil. For this, 91 canine feces samples were collected from 82 farms between August, 2017 and January, 2018. These fecal materials were processed using the techniques of spontaneous sedimentation, centrifugation-floatation in zinc sulfate and Ziehl-Neelsen staining. A questionnaire in the form of an interview was administered to the dogs' owner and information about the farm and the main care provided for the dogs was obtained. Among 91 sampled dogs, 27 (29.7%) were positive for at least one parasite species. Ancylostoma was the most prevalent genus (16.5%), followed by Giardia duodenalis (14.3%), Trichuris vulpis (6.6%), Toxocara canis (5.5%), Entamoeba spp. (4.8%), Cryptosporidium spp. (3.3%) and Taeniidae (1.1%). Most dogs do not receive veterinarian care and rarely received antiparasitic treatment. They were free to roam and had free access to animal remains and garbage, which was reflected in the significant associations with the occurrence of parasites that were found. We conclude that rural dogs harbor gastrointestinal parasites, but that their owners are unaware of the risks that these parasites can bring to human health.

Resumo Foi investigada a ocorrência de parasitos gastrintestinais com potencial zoonótico, em cães de área rural do munícipio de Painel, Santa Catarina, Brasil. Para isso, foram colhidas amostras de fezes de 91 cães de 82 propriedades rurais, no período de agosto de 2017 a janeiro de 2018. Este material fecal foi processado pelas técnicas de sedimentação espontânea, centrífugo-flutuação em sulfato de zinco e coloração pelo Ziehl-Neelsen. Um questionário em forma de entrevista foi aplicado aos tutores dos cães e foram obtidas informações sobre a propriedade rural e principais cuidados com os cães. Entre as 91 amostras de fezes, em 27 (29,7%) foi detectada a presença de parasitos gastrintestinais, sendo Ancylostoma o gênero mais prevalente (15 / 16,5%), seguido por Giardia duodenalis (13 / 14,3%), Trichuris vulpis (6 / 6,6%), Toxocara canis (5 / 5,5%), Entamoeba spp. (4 / 4,8%), Cryptosporidium spp. (3 / 3,3%) e Taeniidae (1 / 1,1%). A maioria dos cães não recebia cuidados veterinários ou antiparasitários. Concluiu-se que os cães de área rural albergam parasitos gastrintestinais, porém seus tutores desconhecem os riscos que esses parasitos podem trazer para a saúde humana.

Aislamiento de Acanthamoeba spp. en salas cerradas de un hospital de la provincia de Buenos Aires, Argentina / Isolation of Acanthamoeba spp. in closed rooms of a hospital in Buenos Aires province, Argentina

Resumen Las amebas de vida libre (AVL) son protozoos ubicuos con cuatro géneros patógenos para el ser humano: Acanthamoeba, Naegleria, Balamuthia, y Sappinia. Acanthamoeba puede actuar como reservorio de microorganismos (endosimbiontes), por lo cual, en medio hospitalario, implicaría un riesgo para la transmisión de bacterias, virus y hongos intranosocomiales. Se investigó la presencia de AVL, con énfasis en Acanthamoeba spp., en un hospital pediátrico de la provincia de Buenos Aires, Argentina. Se colectaron 22 muestras de lavamanos e incubadoras en salas de Neonatología y Terapia Intensiva, las que fueron cultivadas a 37 y 42 °C. Los aislados fueron identificados molecularmente. El 63,64% de las muestras presentaron Acanthamoeba spp. Esta investigación representa el primer estudio realizado en la Argentina sobre la detección de Acanthamoeba spp. en salas cerradas de un hospital. Su presencia es una señal de alarma y resulta un blanco útil para investigar posibles reservorios de microorganismos patógenos en ambientes hospitalarios.

Abstract Free-living amoebae (FLA) are ubiquitous protozoa with four pathogenic genera for humans: Acanthamoeba, Naegleria, Balamuthia, and Sappinia. Acanthamoeba can act as a reservoir of microorganisms (endosymbionts), for which reason, in a hospital environment, it would imply a risk for transmission of nosocomial bacteria, viruses and fungi. The presence of AVL, with emphasis on Acanthamoeba spp., was investigated in a pediatric hospital. Twenty-two samples were collected from sinks and incubators in Neonatology and Intensive Care rooms, which were cultured at 37 and 42 °C. The isolates found were molecularly identified. A total of 63.64% of the samples presented Acanthamoeba spp. This research represents the first study in Argentina on the detection of Acanthamoeba spp. in closed rooms of a hospital. Its presence is an alarm signal, and it is a useful target to investigate possible reservoirs of pathogenic microorganisms in hospital environments.

Resumo As amebas de vida livre (AVL) são protozoários ubíquos com quatro gêneros patogênicos para o ser humano: Acanthamoeba, Naegleria, Balamuthia, e Sappinia. Acanthamoeba pode atuar como um reservatório de microrganismos (endossimbiontes), e portanto, em um ambiente hospitalar, representaria um risco de transmissão de bactérias, vírus e fungos intra-nosocomiais. A presença de AVL, com em Acanthamoeba spp. em um hospital pediátrico da província de Buenos Aires, Argentina, foi investigada. Vinte e duas amostras foram coletadas em lavatórios e incubadoras em Salas de Neonatologia e Cuidados Intensivos, cultivadas a 37 e 42 °C. Os isolados foram identificadas molecularmente. Foram encontradas Acanthamoeba spp. em 63,64% das amostras. Esta investigação representa o primeiro estudo realizado na Argentina sobre a detecção de Acanthamoeba spp. em salas fechadas de um hospital. A sua presença é um sinal de alarme e um alvo para investigar possíveis reservatórios de microrganismos patogênicos em ambientes hospitalares.

Factores de riesgo y parasitosis intestinal en niños menores de 10 años de la olla común / Risk factors and intestinal parasitosis in children under 10 years of age from the common pot

Diversos factores de riesgos pueden condicionar las prevalencias de parasitosis intestinales en niños. Como objetivo se propuso determinar los factores de riesgo y parasitosis intestinal en niños menores de 10 años de la Olla Común ubicada en Villa Mara del Triunfo-Perú durate el año 2022. La investigación fue descriptiva-experimental y correlativa con una muestra de 160 niños <10 años de ambos sexos. Como instrumento de recolección se aplicó un cuestionario con preguntas dicotómicas para conocer las prácticas de higiene de los niños y se efectuó la recolección y procesamiento de muestras con técnicas de Solución Salina S.S.F al 85%, Lugol, Método de Kato y Método de concentración-Flotación de Faust. Para analizar los datos se utilizó Microsoft Excel y el software SPSS, estadística descriptica y hallar tablas de frecuencias y porcentajes y Prueba de correlación entre los elementos de riesgo y la frecuencia de signos de los parásitos. Como resultado, el cálculo estadístico con el método de Spearman mostró una concordancia positiva con el coeficiente de correlación 0,725 y con un valor de 0,001<0,05; donde los niveles de factores de riesgo en los infantes mostraron que el 31,3% se encontraban en riesgo bajo con un nivel bajo de síntomas y sin evidencia de parásitos intestinales, el 18,8% en riesgo medio y el 12,5% en riesgo alto y un nivel alto de síntomas y parasitosis intestinal confirmado; la mayoría de los niños se encuentran en un entorno de bajo riesgo. Se hace necesario e indispensable continuar con invesigaciones en zonas adyacentes(AU)

Various risk factors can condition the prevalence of intestinal parasitosis in children. The objective was to determine the risk factors and intestinal parasitosis in children under 10 years of age from the Common Pot located in Villa Mara del Triunfo-Peru during the year 2022. The research was descriptive-experimental and correlative with a sample of 160 children <10 years of both sexes. As a collection instrument, a questionnaire with dichotomous questions was applied to know the hygiene practices of the children and the collection and processing of samples was carried out with techniques of Saline Solution S.S.F at 85%, Lugol's, Kato's Method and Concentration-Flotation Method. of Faust. To analyze the data, Microsoft Excel and the SPSS software were used, descriptive statistics and finding tables of frequencies and percentages and a correlation test between the risk elements and the frequency of signs of the parasites. As a result, the statistical calculation with the Spearman method showed a positive agreement with the correlation coefficient 0.725 and with a p value of 0.001<0.05; where the levels of risk factors in infants showed that 31.3% were at low risk with a low level of symptoms and no evidence of intestinal parasites, 18.8% at medium risk and 12.5% at high risk and a high level of symptoms and confirmed intestinal parasitism; most children are in a low-risk environment. It is necessary and essential to continue with investigations in adjacent areas(AU)

Evaluación y caracterización de la sirtuina 2.3 de Giardia duodenalis (CdSir2.3)

La giardiasis es la enfermedad gastrointestinal de mayor incidencia mundial, causada por el protozoario Giardia duodenalis, para la cual no se cuenta con una vacuna o tratamiento eficiente. En aras de buscar nuevos blancos farmacológicos contra este parásito, se han estudiado las enzimas del metabolismo energético, como las sirtuinas, deacetilasas dependientes del dinucleótido de adenina y nicotinamida (NAD). Previamente se identificó a GdSir2.1 y GdSir2.2 como deacetilasas dependientes de NAD, con localizaciones subcelulares diferentes. En este trabajo se estudió otro candidato a sirtuina (GdSir2.3) mediante herramientas bioinformáticas para la identificación de características típicas de la familia sirtuina en la secuencia del candidato, y experimentales como la obtención de la proteína recombinante 6xHis-GdSir2.3 que demostró actividad deacetilasa dependiente de NAD y que sirvió como antígeno en la producción de los IgY - α -6xHis-GdSir2.3 para la localización subcelular de la proteína endógena en G. duodenalis. Lo anterior concuerda con otros estudios donde se señala a GdSir2.3 como un importante regulador de la enquistación, debido a su aumento de expresión durante esta etapa del ciclo de vida, constituyéndola como un blanco farmacológico promisorio para el control de esta parasitemia.

Giardiasis is the gastrointestinal disease with the highest incidence worldwide, caused by the protozoan Giardia duodenalis, for which there is no vaccine or efficient treatment. In order to find new pharmacological targets against this parasite, energy metabolism enzymes such as sirtuins, deacetylases dependent on the nicotinamide adenine dinucleotide (NAD), have been studied. GdSir2.1 and GdSir2.2 were previously identified as NAD-dependent deacetylases, with different subcellular locations. In this work, another candidate for sirtuin (GdSir2.3) was studied using bioinformatic tools for the identification of typical characteristics of the sirtuin family in the sequence of the candidate; and experimental ones such as obtaining the recombinant protein 6xHis-GdSir2.3 that demonstrated NAD-dependent deacetylase activity; and that it served as an antigen in the production of IgY - α - 6xHis-GdSir2.3 for the subcellular localization of the endogenous protein in G. duodenalis. The foregoing is consistent with other studies where GdSir2.3 is indicated as an important regulator of encyst due to its increased expression during this stage of the life cycle, constituting it as a promising drug target for the control of this parasitaemia.

A giardíase é a doença gastrointestinal de maior incidência no mundo, causada pelo protozoário Giardia duodenalis, para a qual não existe vacina ou tratamento eficaz. Com o objetivo de encontrar novos alvos farmacológicos contra esse parasita, têm sido estudadas enzimas do metabolismo energético, como as sirtuínas, desacetilases dependentes do dinucleotídeo adenina nicotinamida (NAD). GdSir2.1 e GdSir2.2 foram previamente identificados como desacetilases dependentes de NAD, com diferentes localizações subcelulares. Neste trabalho, outro candidato a sirtuin (GdSir2.3) foi estudado usando ferramentas de bioinformática para a identificação de características típicas da família sirtuin na sequência do candidato; e experimentais, como a obtenção da proteína recombinante 6xHis-GdSir2.3 que demonstrou atividade desacetilase dependente de NAD; e que serviu como antígeno na produção de IgY - α - 6xHis-GdSir2.3 para a localização subcelular da proteína endógena em G. duodenalis. O exposto é consistente com outros estudos em que o GdSir2.3 é apontado como um importante regulador de encisto devido à sua expressão aumentada durante esta fase do ciclo de vida, constituindo-se como um alvo promissor para o controle dessa parasitemia.

Frecuencia y distribución de enteroparásitos en área rural del departamento de La Paz / Frequency and distribution of enteroparasites in rural area in La Paz

INTRODUCCIÓN: las enfermedades parasitarias representan un problema de salud pública debido a su alta prevalencia por todo el mundo sobre todo en países en desarrollo, especialmente en áreas rurales y Bolivia no es la excepción. OBJETIVO: el presente estudio tuvo el objetivo de determinar la frecuencia y distribución de enteroparásitos en 8 municipios rurales del departamento de La Paz durante el periodo de agosto a septiembre de 2014. MATERIALES Y MÉTODOS: el trabajo es un estudio tipo transversal-descriptivo con un universo de trabajo de 1238 muestras de heces fecales conservadas con formol, las cuales fueron enviadas por el equipo médico de SUYANA (organización no gubernamental sin fines de lucro) a los laboratorios del Instituto SELADIS (Servicios de Laboratorio de Diagnóstico e Investigación en Salud). RESULTADOS: se realizaron estudios coproparasitológicos directos de cada una de las muestras y la observación microscópica dio los siguientes resultados. Se evidencio la presencia de enteroparásitos en 89,5% de la población estudiada, de los cuales 97,2% representan protozoarios tales como B. hominis, E. coli (protoozoos comensales) y G. lamblia (protozoo intestinal patógeno) los cuales estarían como los de mayor distribución, por otro lado el 2.8% de la población total corresponden a helmintos donde H. nana tiene una distribución de 1,8%, A. lumbricoides 0,7% y T. trichiura, S. stercoralis, Uncinarias estarían en el 0,1% de la población. También se pudo evidenciar que la mayor distribución de enteroparásitos estaría entre 1-10 años de edad (37,4%). Finalmente se evidencio que de las 8 poblaciones de estudio Charazani, Calacoto, Comanche presentarían mayor distribución de enteroparásitos (10-11 parásitos). CONCLUSIÓN: se evidencio que casi el 90% de la población en estudio, presentarían parásitos intestinales, con un claro predominio de los protozoarios sobre los helmintos. Esta información epidemiológica servirá de apoyo para mejorar los programas de salud en estas poblaciones.

INTRODUCTION: parasitic diseases represent a public health problem because of its high prevalence throughout the world, especially in developing countries, particularly in rural areas and Bolivia is no exception. OBJETIVE: therefore, the present study had the objective of determining the frequency and distribution of enteroparasites in eight municipalities of La Paz between August and September in 2014. MATERIALS AND METHODS: this was a cross-sectional-descriptive study with a universe of 1238 formalin-preserved stool samples, which were sent for analysis by the medical team of SUYANA (a non-profi t organization) to the Bolivian Institute of Health Diagnostic and Research Laboratory Services (SELADIS, acronym in Spanish). RESULTS: Copro-parasitological studies of each sample and microscopic analysis were detected. As a result, prevalence of infection by any given enteroparasites were detected, 89.5% of protozoans (B. hominis, E. coli (commensal protozoa) and 97.2%, G. lamblia (pathogenic intestinal protozoa) which represent the highest distribution. Besides, we identified, 2.8% of helmintos, among them, 1.8% of H. nana, 1.7% of A. lumbricoides and 0.1% of others (T. trichiura, S. stercoralis and Uncinarias). According to age group, 37.4% were people between 1 and 10 years old (enteroparasites infection). From eight municipalities, Charazani, Calacoto, and Comanche had the highest distribution of enteparasites (10-11 parasites). CONCLUSIONS: the present study showed that 90% of the population had intestinal parasites, where protozoans were higher than helminths. This epidemiological information could be reliable to improve health care programs for these populations.

Mortalidad por parasitosis endémicas e importadas en Chile. 1997-2020 / Mortality due to endemic and imported parasitoses in Chile. 1997-2020

INTRODUCCIÓN: Las parasitosis son un problema de salud pública mundial por su alta prevalencia en países en vías de desarrollo, su persistencia en países desarrollados debido principalmente por la migración de personas, y por su morbi-mortalidad asociada. OBJETIVO: Determinar las defunciones causadas por parasitosis endémicas e importadas en Chile, según región, sexo, grupo etario, procedencia urbana/rural y nivel educacional, y estimar tasas de mortalidad. MÉTODO: Estudio de base poblacional y serie de tiempo de mortalidad (1997 a 2020). Los análisis estadísticos fueron realizados mediante programas Excel y R Studio, cálculo de χ2, valor p < 0,05. RESULTADOS: 2.413 muertes fueron causadas por parásitos, 65,1% por protozoos, 33% por helmintos y 1,9% por artrópodos. La tasa promedio de mortalidad nacional fue de 0,6 por cada 100 mil habitantes. Las principales causas de fallecimiento fueron: enfermedad de Chagas (63,6%), hidatidosis (24,3%) y cisticercosis (8,1%). La mayoría de los fallecidos pertenecía a la Región de Coquimbo. La edad promedio de defunción fue de 74, 62 y 67 años para muertes por protozoos, helmintos y artrópodos, respectivamente. Las defunciones ocurrieron mayormente en hombres. La mayoría provenía de zona urbana y poseían un nivel educacional bajo. CONCLUSIONES: La enfermedad de Chagas es la principal causa de muerte por parasitosis en Chile.

BACKGROUND: Parasites are a public health problem due to their high prevalence in developing countries, their persistence in developed countries mainly due to the migration of people, and their associated morbidity and mortality. AIM: To determine the deaths in Chile caused by endemic and imported parasites; according to region, sex, age group, urban-rural origin and educational level, and mortality rates. METHOD: Population-based study, times series of mortality (1997 to 2020). Statistical analysis was performed using Excel and R Studio programs, calculation of χ2, p value < 0.05. RESULTS: 2,413 deaths occurred due to parasites, 65.1% by protozoa, 33% by helminths and 1.9% by arthropods. The average national mortality rate was 0.6 per 100.000 inhabitants. The main causes of death were Chagas disease (63.6%), hydatidosis (24.3%) and cysticercosis (8.1%). Most of the deceased belonged to the Coquimbo Region. The mean age of death was 74, 62 and 67 years old for deaths from protozoa, helminths and arthropods, respectively. Deaths occurred mostly in men. Most came from urban areas and had a low educational level. CONCLUSIONS: Chagas disease is the main cause of death from parasites in Chile.

Detection of enteroparasites in foliar vegetables commercialized in street- and supermarkets in Aparecida de Goiânia, Goiás, Brazil / Detecção de enteroparasitas em hortaliças comercializadas em feiras e supermercados em Aparecida de Goiânia, Goiás, Brasil

Foliar vegetables contaminated with fecal residues are an important route of transmission of intestinal parasites to humans. The aim of this study was to evaluate the presence of parasitic forms of protozoa and helminths on lettuces (Lactuca sativa) and collard greens (Brassica oleracea) sold in street- and supermarkets in the city of Aparecida de Goiânia, Goiás, Brazil. A total of 30 samples of each vegetable (15 samples from each supermarkets and street markets) was analyzed. All samples were processed by spontaneous sedimentation method and centrifugal flotation. In 45% of the samples, immature forms of intestinal parasites were identified with 66.7% helminths eggs and 33.3% protozoan cysts or oocysts. Significantly more lettuce samples were contaminated with eggs, cysts or oocyst of at least one parasite than collard green samples (U=216; Z=-3.45; P <0.001). The parasitic forms were identified morphologically up to the family level with eggs of Ancylostomatidae, Strongyloididae, Ascarididae and Taeniidae, or oocysts of Eimeriidae, to the genus with Cystoisospora sp. and Toxocara sp., and to the species level with Cystoisospora canis, Dipylidium caninum and Hymenolepis nana. The presence of these infective agents in lettuce and collard green from both street- and supermarkets highlights the high risk of spreading parasites by eating raw vegetables sold in Aparecida de Goiânia.

Hortaliças contaminadas com resíduos fecais são importantes vias de transmissão de parasitas intestinais ao homem. O objetivo deste estudo foi avaliar a presença de formas parasitárias de protozoários e helmintos em alface (Lactuca sativa) e couve (Brassica oleracea) vendidas em feiras e supermercados na cidade de Aparecida de Goiânia, Goiás, Brasil. Um total de 30 amostras de cada vegetal (15 amostras de supermercados e 15 de feiras livres) foi analisado. Todas as amostras foram processadas pelos métodos de sedimentação espontânea e centrífugo-flutuação. Em 45% das amostras foram identificadas formas imaturas de parasitas intestinais sendo 66,7% ovos de helmintos e 33,3% de cistos ou oocistos de protozoários. Significativamente, mais amostras de alface estavam contaminadas com ovos, cistos ou oocistos de pelo menos um parasita do que as amostras de couve (U = 216; Z = -3,45; P <0,001). As formas parasitárias foram identificadas morfologicamente ao nível de família com ovos de Ancylostomatidae, Strongyloididae, Ascarididae e Taeniidae ou oocistos de Eimeriidae, dos gêneros Cystoisospora sp. e Toxocara sp., e ao nível de espécies com Cystoisospora canis, Dipylidium caninum e Hymenolepis nana. A presença desses agentes infecciosos em alface e couve, provenientes tanto de feiras quanto de supermercados, ressalta o alto risco de veiculação de parasitas pela ingestão de hortaliças cruas comercializadas em Aparecida de Goiânia.

Standardization of molecular techniques for the detection and characterization of intestinal protozoa and other pathogens in humans

Background: The intrinsic sensitivity limitations of basic parasitological methods, along with the particular biological characteristics of parasites, make these methods ineffective to differentiate morphologically indistinguishable species. Molecular detection and characterization techniques could be used to overcome these problems. The purpose of this work was to standardize molecular polymerase chain reaction (PCR) techniques, described in the literature, for the detection and molecular characterization of intestinal protozoa and other pathogens in humans. Methods: DNA was extracted from human or animal feces, previously washed or cultured in Boeck Drbohlav's Modified Medium. DNA extraction was performed with Machery-Nagel extraction kits. The standardization of the PCR, nested-PCR or RFLP techniques was carried out according to the literature. For each molecular technique performed, the sensitivity of the test was determined based on the minimun quantity required of DNA (sensitivity A) and the minimum quantity of life forms that the test detected (sensitivity B). Results: Sensitivity A was 10 fg for G. duodenalis, 12.5 pg for Entamoeba histolytica or Entamoeba dispar, 50 fg for Cryptosporidium spp., 225 pg for Cyclospora spp. and 800 fg or 8 fg for Blastocystis spp. after performing a 1780 bp PCR or 310 bp nested PCR, respectively. The sensitivity B was 100 cysts for G. duodenalis, 500 cysts for E. histolytica or E. dispar, 1000 oocysts for Cyclospora spp. and 3600 or four vegetatives forms for PCR or nested PCR of Blastocystis spp., respectively. Conclusions: The molecular detection of protozoa and chromist was achieved and the molecular characterization allowed the genotyping of some of the parasites such as Giardia duodenalis, Cryptosporidium spp., and Blastocystis spp. This study summarizes the molecular techniques for epidemiological studies in humans and animals, and helps in the investigation of their transmission sources in countries where intestinal parasites are a public health problem.(AU)

Detection of enteroparasites in foliar vegetables commercialized in street- and supermarkets in Aparecida de Goiânia, Goiás, Brazil / Detecção de enteroparasitas em hortaliças comercializadas em feiras e supermercados em Aparecida de Goiânia, Goiás, Brasil

Foliar vegetables contaminated with fecal residues are an important route of transmission of intestinal parasites to humans. The aim of this study was to evaluate the presence of parasitic forms of protozoa and helminths on lettuces (Lactuca sativa) and collard greens (Brassica oleracea) sold in street- and supermarkets in the city of Aparecida de Goiânia, Goiás, Brazil. A total of 30 samples of each vegetable (15 samples from each supermarkets and street markets) was analyzed. All samples were processed by spontaneous sedimentation method and centrifugal flotation. In 45% of the samples, immature forms of intestinal parasites were identified with 66.7% helminths eggs and 33.3% protozoan cysts or oocysts. Significantly more lettuce samples were contaminated with eggs, cysts or oocyst of at least one parasite than collard green samples (U=216; Z=-3.45; P <0.001). The parasitic forms were identified morphologically up to the family level with eggs of Ancylostomatidae, Strongyloididae, Ascarididae and Taeniidae, or oocysts of Eimeriidae, to the genus with Cystoisospora sp. and Toxocara sp., and to the species level with Cystoisospora canis, Dipylidium caninum and Hymenolepis nana. The presence of these infective agents in lettuce and collard green from both street- and supermarkets highlights the high risk of spreading parasites by eating raw vegetables sold in Aparecida de Goiânia.

Hortaliças contaminadas com resíduos fecais são importantes vias de transmissão de parasitas intestinais ao homem. O objetivo deste estudo foi avaliar a presença de formas parasitárias de protozoários e helmintos em alface (Lactuca sativa) e couve (Brassica oleracea) vendidas em feiras e supermercados na cidade de Aparecida de Goiânia, Goiás, Brasil. Um total de 30 amostras de cada vegetal (15 amostras de supermercados e 15 de feiras livres) foi analisado. Todas as amostras foram processadas pelos métodos de sedimentação espontânea e centrífugo-flutuação. Em 45% das amostras foram identificadas formas imaturas de parasitas intestinais sendo 66,7% ovos de helmintos e 33,3% de cistos ou oocistos de protozoários. Significativamente, mais amostras de alface estavam contaminadas com ovos, cistos ou oocistos de pelo menos um parasita do que as amostras de couve (U = 216; Z = -3,45; P <0,001). As formas parasitárias foram identificadas morfologicamente ao nível de família com ovos de Ancylostomatidae, Strongyloididae, Ascarididae e Taeniidae ou oocistos de Eimeriidae, dos gêneros Cystoisospora sp. e Toxocara sp., e ao nível de espécies com Cystoisospora canis, Dipylidium caninum e Hymenolepis nana. A presença desses agentes infecciosos em alface e couve, provenientes tanto de feiras quanto de supermercados, ressalta o alto risco de veiculação de parasitas pela ingestão de hortaliças cruas comercializadas em Aparecida de Goiânia.

Detection of enteroparasites in foliar vegetables commercialized in street- and supermarkets in Aparecida de Goiânia, Goiás, Brazil

Abstract Foliar vegetables contaminated with fecal residues are an important route of transmission of intestinal parasites to humans. The aim of this study was to evaluate the presence of parasitic forms of protozoa and helminths on lettuces (Lactuca sativa) and collard greens (Brassica oleracea) sold in street- and supermarkets in the city of Aparecida de Goiânia, Goiás, Brazil. A total of 30 samples of each vegetable (15 samples from each supermarkets and street markets) was analyzed. All samples were processed by spontaneous sedimentation method and centrifugal flotation. In 45% of the samples, immature forms of intestinal parasites were identified with 66.7% helminths eggs and 33.3% protozoan cysts or oocysts. Significantly more lettuce samples were contaminated with eggs, cysts or oocyst of at least one parasite than collard green samples (U=216; Z=-3.45; P 0.001). The parasitic forms were identified morphologically up to the family level with eggs of Ancylostomatidae, Strongyloididae, Ascarididae and Taeniidae, or oocysts of Eimeriidae, to the genus with Cystoisospora sp. and Toxocara sp., and to the species level with Cystoisospora canis, Dipylidium caninum and Hymenolepis nana. The presence of these infective agents in lettuce and collard green from both street- and supermarkets highlights the high risk of spreading parasites by eating raw vegetables sold in Aparecida de Goiânia.

Resumo Hortaliças contaminadas com resíduos fecais são importantes vias de transmissão de parasitas intestinais ao homem. O objetivo deste estudo foi avaliar a presença de formas parasitárias de protozoários e helmintos em alface (Lactuca sativa) e couve (Brassica oleracea) vendidas em feiras e supermercados na cidade de Aparecida de Goiânia, Goiás, Brasil. Um total de 30 amostras de cada vegetal (15 amostras de supermercados e 15 de feiras livres) foi analisado. Todas as amostras foram processadas pelos métodos de sedimentação espontânea e centrífugo-flutuação. Em 45% das amostras foram identificadas formas imaturas de parasitas intestinais sendo 66,7% ovos de helmintos e 33,3% de cistos ou oocistos de protozoários. Significativamente, mais amostras de alface estavam contaminadas com ovos, cistos ou oocistos de pelo menos um parasita do que as amostras de couve (U = 216; Z = -3,45; P 0,001). As formas parasitárias foram identificadas morfologicamente ao nível de família com ovos de Ancylostomatidae, Strongyloididae, Ascarididae e Taeniidae ou oocistos de Eimeriidae, dos gêneros Cystoisospora sp. e Toxocara sp., e ao nível de espécies com Cystoisospora canis, Dipylidium caninum e Hymenolepis nana. A presença desses agentes infecciosos em alface e couve, provenientes tanto de feiras quanto de supermercados, ressalta o alto risco de veiculação de parasitas pela ingestão de hortaliças cruas comercializadas em Aparecida de Goiânia.

Taxonomic rank of human parasites / 中国血吸虫病防治杂志

Biological category is effective to indicate the evolution of organism populations between past and present. Conventional taxonomy of human parasites mainly depends on important morphological features, which suffers from a problem of categorizing related-genera species with similar morphological characteristics. With recent advances in molecular biological technologies, the effective applications of mitochondrial and ribosomal biomarkers and sequencing greatly improve the development of the taxonomic rank of human parasites. Worldwide, the classification of human parasites have been continuously revised and improved. Hereby, we re-categorize parasitic Protozoa, Trematoda, Cestoda and Nematoda, so as to provide insights into the researches on molecular systematics and genetic evolution of human parasites.

Balantidium coli apendicular como hallazgo incidental. Reporte de un caso / Apendicular balantidium coli as incidental finding during surgery. Report of a case

Resumen Balantidium coli es el único miembro de la familia Balantidiidae capaz de producir infección en seres humanos. Presentamos un caso en un hombre de 43 años que ingresa para corrección quirúrgica de hernia ventral durante la cual se realizó apendicetomía profiláctica. En el estudio histopatológico se observó apéndice cecal con arquitectura conservada, sin la presencia de apendicitis ni periapendicitis. En la luz se reconocieron estructuras grandes (aproximado de 50 μm) redondas con citoplasma amplio con vacuolas grandes, cilias periféricas y núcleos densos, los cuales correspondieron a trofozoitos de Balantidium coli.

Abstract Balantidium coli is the only member of the Balantidiiae family capable of infecting human beings. We present one in a 43 years-old male admitted for a surgical co rrection of an incisional hernia with prophylactic appendicectomy. Histopathological findings reported the cecal appendix within normal architecture, appendicitis and peri-appendicitis free. At the lumen big, rounded shape structures (aprox. 50 mm) were visible with broad cytoplasm, big vacuoles, peripheral cilia and dense nucleus, corresponding to Balantidium coli trophozoites.

Eimeria bateri Natural Infection: Oocysts Reductions in Grey Quail (Coturnix coturnix) Treated with Bacillus thuringensis var. israelensis

Abstract Coccidiosis, a disease caused by the parasitic Eimeria spp., affects birds of all ages, particularly young birds more intensely. Infected poultry presents significant economic losses. Bacillus thuringiensis var israelensis (Bti) is a Gram-positive, spore-forming bacterium that produces proteins with high specific parasiticidal activity against various orders of parasites. Thus, the aim of the present study was to evaluate the parasiticidal potential of Bti in quails that were naturally infected with Eimeria bateri. Twenty 12-week-old male quails (Coturnix coturnix coturnix), naturally infected with Eimeria bateri, were randomly divided into two groups of 10 birds: Bti treated and control. The treated group was supplemented with Bti (1×108 spores∙g-1) in the feed, while; the control group received the same feed without Bti. To evaluate the occurrence of oocysts, samples of feces were collected every week for four weeks. Significant (P < 0.05) oocysts reductions of 56.64% and 94.51% were noted in the Bti treated group at 2nd and 4th week of study, respectively. The Bti supplementation may contribute to the reduction of oocysts in quails and environmental contamination. Bacillus thuringiensis var israelensis appeared to be a promising complementary alternative in E. bateri control.

Endoparasites of capybaras (Hydrochoerus hydrochaeris) from anthropized and natural areas of Brazil / Endoparasitas de capivaras (Hydrochoerus hydrochaeris) provenientes de áreas antropizadas e de áreas naturais do Brasil

Abstract This study aimed to evaluate the occurrence of endoparasites in fecal samples of capybaras from anthropized areas in seven cities in the state of São Paulo, southeastern Brazil, and natural areas in two locations in the Pantanal biome. For the parasitological evaluation of samples, feces were subjected to centrifugal sedimentation in water-ether and centrifugal flotation in sucrose solution. Out of the 113 samples examined, 97.3% (110/113) were positive for the presence of helminth eggs and/or protozoan oocysts, with 96.7% (89/92) and 100% (21/21) in capybaras from anthropized and natural areas, respectively. Helminth eggs belonging to the Class Cestoda (Monoecocestus spp.), Class Digenea, and Class Nematoda (Trichostrongyloidea, Strongyloides chapini, Protozoophaga obesa, and Capillaria hydrochoeri) were detected. Coccidia oocysts of Eimeria spp. and Cryptosporidium spp. were detected. The difference found was in the occurrence of S. chapini between anthropized and natural areas, as S. chapini was not detected in natural areas. Capybaras from anthropized areas showed a greater species richness of endoparasites, including species such as Fasciola hepatica and Cryptosporidium spp., that might have zoonotic potential.

Resumo O presente estudo apresentou como objetivo avaliar a ocorrência de endoparasitas nas amostras de fezes de capivaras de áreas antropizadas de sete cidades do Estado de São Paulo, Sudeste do Brasil, e áreas naturais de duas localidades do bioma Pantanal. Para a avaliação parasitológica das amostras de fezes foram empregadas as técnicas de centrífugo-sedimentação em água-éter e centrífugo-flutuação em solução de sacarose. De um total de 113 amostras examinadas, 97,3% (110/113) apresentaram resultados positivos para a presença de ovos de helmintos e/ou oocistos de protozoários. No geral, a positividade foi de 96,7% (89/92), e 100% (21/21) em capivaras de áreas antropizadas e naturais, respectivamente. Foram detectados ovos de helmintos pertencentes às Classes Cestoda (Monoecocestus spp.), Digenea e Nematoda (Trichostrongyloidea, Strongyloides chapini, Protozoophaga obesa e Capillaria hydrochoeri). Em relação aos protozoários, foram detectados oocistos dos coccídios Eimeria spp. e Cryptosporidium spp. A diferença encontrada foi na comparação da ocorrência de S. chapini em áreas antropizadas e naturais, sem detecção para esta última área. As capivaras provenientes das áreas antropizadas apresentaram uma maior riqueza de espécies de endoparasitas, incluindo as espécies Fasciola hepatica e Cryptosporidium spp., que podem apresentar potencial zoonótico.

Bioinformatics of thymidine metabolism in Trypanosoma evansi: exploring nucleoside deoxyribosyltransferase (NDRT) as a drug target

@#Trypanosoma evansi, the causative agent of surra or camel trypanosomiasis, is characterized by the widest geographic distribution and host range among the known trypanosomes. Its zoonotic importance and increasing evidence of drug resistance necessitate the discovery of new drug targets. The drug discovery process entails finding an exploitable difference between the host and the parasite. In this study, the thymidine metabolic pathways in camel and T. evansi were compared by analyzing their metabolic maps, protein sequences, domain and motif contents, phylogenetic relationships, and 3D structure models. The two organisms were revealed to recycle thymidine differently: performed by thymidine phosphorylase in camels (Camelus genus), this role in T. evansi was associated with nucleoside deoxyribosyltransferase (NDRT), a unique trypanosomal enzyme absent in camels. Thymidine in T. evansi seems to be governed by thymine through NDRT, whereas in camels, thymidine can be produced from thymidylate via 5'-nucleotidase. As a result, NDRT may be a promising drug target against T. evansi.

Occurrence and molecular characterization of Giardia duodenalis from naturally infected dogs in the municipality of Santa Maria, Rio Grande do Sul, Brazil / Ocorrência e caracterização molecular de Giardia duodenalis em cães naturalmente infectados no município de Santa Maria, Rio Grande do Sul, Brasil

Giardiasis is an important and prevalent zoonosis in dogs and humans caused by Giardia spp. The close relationship between pets and humans has physical, emotional and social benefits. The dogs have an important role in Giardia duodenalis cycle and transmission. This study aimed to verify the occurrence of the parasite in dogs from Central Region, in Santa Maria, Rio Grande do Sul State, Brazil, from April to October 2018. Dog feces (230) were submitted to Faust coproparasitological and molecular analyses. The positive samples in the nested-PCR (β-giardin gene) were sent for DNA sequencing and phylogenetic analyses (Neighbor-Joining). The occurrence of G. duodenalis, was 5.6% (13/230) and 4.3% (10/230) detected by coproparasitological technique and nested-PCR, respectively. There was no difference in the sensitivity of the tests used. From the faecal samples analyzed, there were no differences among the variables: diagnostic techniques, local, sex, and age of the animals (p>0.05). Only in the stool examination methodology a difference was observed between the ages (p<0.05). G. duodenalis assemblages were C and D, frequently reported in dogs. The close relationship between dogs and people may allow co-infections of circulating parasites in the population, including Giardia spp. and increasing the risk of transmission of zoonotic agents.(AU)

A giardíase é uma zoonose importante e prevalente em cães e humanos, sendo causada por Giardia spp. A estreita relação entre animais de estimação e seres humanos traz benefícios físicos, emocionais e sociais. Os cães têm um papel importante no ciclo e transmissão de Giardia duodenalis. Este estudo teve como objetivo verificar a ocorrência do parasita em cães da Região Central, em Santa Maria, RS, Brasil, de abril a outubro de 2018. As fezes de cães (230) foram submetidas a técnica coproparasitológica de Faust e análises moleculares. As amostras positivas no nested-PCR (gene β-giardin) foram enviadas para sequenciamento de DNA e posterior análise filogenética (Neighbor-Joining). A ocorrência de G. duodenalis foi de 5,6% (13/230) e 4,3% (10/230) detectados pela técnica coproparasitológica e nested-PCR, respectivamente. Não houve diferença na sensibilidade dos testes utilizados. Das amostras fecais analisadas, não houve diferenças entre as variáveis: técnicas de diagnóstico, local, sexo e idade dos animais (p>0,05). Somente na metodologia de exame de fezes observou-se diferença entre as idades (p<0,05). As assemblages de G. duodenalis encontradas foram C e D, frequentemente relatadas em cães. A estreita relação entre cães e pessoas pode permitir co-infecções de parasitas circulantes na população, incluindo Giardia spp. e aumentando o risco de transmissão de agentes zoonóticos.(AU)

Cloning and expression of Neospora caninum rhoptry 2 and its potential as an immunobiological for neosporosis control / Clonagem e expressão de rhoptry 2 Neospora caninum e seu potencial como um imunobiológico para o controle de neosporose

The protozoan Neospora caninum is known worldwide as one of the main causes of abortion in cattle. During infection, rhoptry proteins present in the apical complex of the parasite play important roles in adhesion and parasitophorous vacuole formation. The use of N. caninum ROP2 in experimental vaccines has shown promising protective results. In our study we performed cloning and expression in Escherichia coli of an antigenic portion of N. caninum ROP2. The recombinant protein (rROP2) was obtained in insoluble form, and the purified protein showed a size of approximately 18kDa. Even being a small truncate NcROP2 region, it was possible to conserve the antigenic epitopes which were recognized by bovine serum naturally infected with N. caninum. Vaccination with rROP2 on aluminum hydroxide adjuvant induced high levels of rROP2-specific IgG antibodies capable of recognizing native protein in tachyzoite lysates. In conclusion, our approaches were effective in obtaining the rROP2 protein, which induced specific mouse immune response and was also recognized by sera from N. caninum naturally infected cattle. These results suggest that it is a promising antigen for the development of neosporosis subunit vaccines as well as a suitable antigen for use in immunodiagnosis.(AU)

O protozoário Neospora caninum é conhecido mundialmente como uma das principais causas de aborto em bovinos. Durante a infecção, as proteínas rhoptry presentes no complexo apical do parasita desempenham papel importante na adesão e formação de vacúolos parasitóforos. O uso de ROP2 de N. caninum em vacinas experimentais tem mostrado resultados de proteção promissores. Em nosso estudo, realizamos a clonagem e expressão em Escherichia coli de uma porção antigênica de N. caninum ROP2. A proteína recombinante (rROP2) foi obtida na forma insolúvel, e a proteína purificada apresentou tamanho aproximado de 18kDa. Mesmo sendo uma pequena região truncada de NcROP2, foi possível conservar os epítopos antigênicos que foram reconhecidos pelo soro de bovinos naturalmente infectados com N. caninum. A vacinação com rROP2 adsorvida no adjuvante de hidróxido de alumínio induziu altos níveis de anticorpos IgG anti-rROP2, capazes de reconhecer a proteína nativa em lisados de taquizoítos. Em conclusão, nossas abordagens foram eficazes na obtenção da proteína rROP2, que induziu resposta imune específica em camundongos e também foi reconhecida por soros de bovinos naturalmente infectados com N. caninum. Estes resultados sugerem que rROP2 é um antígeno promissor para o desenvolvimento de vacinas de subunidades de neosporose, bem como um antígeno adequado para uso em imunodiagnóstico.(AU)

Molecular detection of Apicomplexa protozoa in tissues from Alouatta guariba clamitans / Detecção molecular de protozoários Apicomplexa em tecidos de Alouatta guariba clamitans

The brown howler monkey (Alouatta guariba clamitans) is a primate species widely distributed in South America. Infections by protozoa are common in primates. However, studies on protozoa in primates in Brazil are scarce, so the goal of this study was to investigate DNA from the apicomplexan protozoa Neospora caninum, Sarcocystis spp. and Toxoplasma gondii in tissues of A. guariba clamitans. DNA extraction was performed on tissue samples from the heart, brain, liver, spleen, lung and intestine of six A. guariba clamitans from Santa Maria, Central Region of Rio Grande do Sul, Brazil. Conventional PCR was performed using 18S rRNA gene general primers for Apicomplexa and also specific primers to amplify Neosporaspp. and Toxoplasma gondii DNA. All animals were positive in the 18S PCR and the genetic sequencing confirmed the presence of Sarcocystis spp. DNA in the tissues of four animals belonging to at least two species (S. neurona and S. gigantea) and T. gondii DNA in the other two animals. One positive sample for T. gondii was genotypically characterized as atypical by the restriction fragment length polymorphism technique. N. caninum DNA was not detected in the tested samples. The presence of Apicomplexa protozoan DNA in the tissues of the six animals tested in this study highlights the importance of howler monkeys as maintainers of these pathogens in nature.(AU)

O bugio ruivo (Alouatta guariba clamitans) é uma espécie de primata amplamente distribuída na América do Sul. As infecções por protozoários são comuns em primatas. Entretanto, estudos sobre protozoários em primatas no Brasil são escassos, portanto o objetivo deste estudo foi pesquisar DNA dos protozoários Apicomplexa Neospora caninum, Sarcocystisspp. e Toxoplasma gondii em tecidos de A. guariba clamitans. A extração de DNA foi realizada em amostras de tecido do coração, cérebro, fígado, baço, pulmão e intestino de seis A. guariba clamitans oriundos de Santa Maria, Região Central do Rio Grande do Sul, Brasil. Foi realizada PCR convencional utilizando primers geral do gene 18S rRNA para Apicomplexa e também primers específicos para amplificação de DNA de Neospora spp.e Toxoplasma gondii. Todos os animais foram positivos no PCR geral para Apicomplexa e no sequenciamento genético confirmou-se a presença de DNA de Sarcocystis nos tecidos de quatro animais pertencentes a pelo menos duas espécies (S. neurona e S. gigantea), e DNA de T. gondii foi detectado nos outros dois animais. Uma amostra positiva para T. gondii foi caracterizada genotipicamente como atípico pela técnica de polimorfismo do comprimento do fragmento de restrição. Não foi detectado DNA de N. caninum nas amostras testadas. A presença de DNA de protozoários apicomplexa nos tecidos dos seis animais testados neste estudo destaca a importância dos bugios ruivos como mantenedores desses patógenos na natureza.(AU)

Epidemiological study of human intestinal parasites in Sarawak, East Malaysia: A review

@#Intestinal parasitic infections are endemic in rural settings and may account for asymptomatic infections to various health complications. These infections are a cause of concern for communities of lower economic status, especially in developing countries. In Sarawak, indigenous populations residing in geographically inaccessible areas are socially and economically disadvantaged. Through close association with nature, these populations are prone to intestinal parasitism. Currently, scattered information has led to a continual state of neglect at each level of parasitic infection control. This urges for a review of their distribution and transmission based on previous reports to understand the pattern of the diseases in the state which can further address the improvement of mass controlling programs. A literature search was conducted to collect previous reports on human intestinal parasites in Sarawak, East Malaysia from PubMed (Medline), SCOPUS, ScienceDirect and Web of Science from January 2019 to March 2021. Extrapolating the current data in Sarawak which is still considered limited, further interdisciplinary strategies are demanded to give insights in the epidemiology and true prevalence of intestinal parasites in Sarawak. This review addresses for redirection of attitude towards intestinal parasitic infections where it should be given with ample attention by rural populations. In tandem to that, improvement of rural livelihood such as standard of living and sanitation in Sarawak should be accredited as part of the efforts to reduce the number of intestinal parasitic infections in the state. As a control measure, mass deworming should be reconsidered especially to the rural populations.