Dogs naturally infected by Rangelia vitalii, Babesia canis vogeli, and Ehrlichia canis in São Paulo, Brazil / Cães naturalmente infectados por Rangelia vitalii, Babesia canis vogeli, e Ehrlichia canis em São Paulo, Brasil

Several agents can cause hemoparasitic diseases in dogs, and blood-sucking arthropods transmit these diseases. These agents can cause several clinical manifestations and, in some cases, can kill the host. Because these agents are essential in animal health, this study aims to detect the frequency of Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, and Rangelia vitalii by real-time PCR and Babesia vogeli in dogs in the southern region of the city of São Paulo, São Paulo. Of the 98 dog samples, 18 (18.4%) tested positive with real-time polymerase chain reaction for at least one studied agent. Of these 18 samples, 17 tested positive for a single agent (11.2% for B. canis vogeli, 1.02% for R. vitalii, and 5.1% for E. canis), and one showed co-infection with B. canis vogeli and R. vitalii. The results demonstrate the presence of hemoparasites in the studied animals, which can influence the quality and life expectancy of these animals. The Rangeliadetection warns small animal clinicians to include it as a differential diagnosis for hemoparasitosis.(AU)

As hemoparasitoses em cães podem ser causadas por diversos agentes, sendo essas doenças transmitidas por artrópodes hematófagos. Esses agentes podem causar diversas manifestações clínicas e, em alguns casos, podem matar o hospedeiro. Este estudo teve como objetivo detectar por PCR em tempo real a frequência de Ehrlichia canis, Rickettsia rickettsii, Anaplasma platys, Rangelia vitalii e Babesia canis vogeli em amostras de cães da zona sul da cidade de São Paulo, Brasil. Das 98 amostras de cães, 18 (18,4%) testaram positivo com reação em cadeia da polimerase em tempo real para pelo menos um agente estudado. Destas 18 amostras, 17 testaram positivo para um único agente (11,2% para B. canis vogeli, 1,02% para R. vitalii e 5,1% para E. canis), e uma apresentou coinfecção com B. canis vogeli e R. vitalii. Os resultados demonstram a presença de hemoparasitas nos animais estudados, o que pode influenciar a qualidade e a expectativa de vida desses animais. Além disso, é o primeiro relato da detecção de R. vitalli na zona sul de São Paulo e serve de alerta para os clínicos de pequenos animais incluírem esse agente como diagnóstico diferencial para as hemoparasitoses.(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)

Urbanorum spp novo parasita no Brasil / Urbanorum spp new parasite in Brazil / Urbanorum spp nuevo parásito en Brasil

Introdução: As enteroparasitoses são foco de investigações científicas no mundo todo. Urbanorumspp. foi reconhecido como parasita em 1994 no Peru, expandindo-se pela América do Sul. Relatado pela primeira vez no Brasil em 2018, Maranhão. Este relato apresenta o segundo caso no estado do Paraná. Relato de caso: Paciente masculino, 56 anos, 75kg, diabético, habitante de São José dos Pinhais, área urbana. Procura atenção primária por dor ao evacuar, tenesmo e cólica abdominal. Nega diarréia, febre, sangue nas fezes e viagem recente. Exame físico abdominal, hemograma e parcial de urina sem alterações. Parasitológico de fezes: Urbanorum spp. Prescrito Nitazoxanida 500mg 12/12h por 3 dias. Paciente retorna com melhora da sintomatologia e parasitológico de controle negativo. Conclusão: Atualmente a escassez de estudos primários prospectivos dificultam o delineamento clínico-epidemiológico e tratamento da parasitose. A disseminação do parasita entre extremos do país em curto intervalo de tempo, aliada à carência de saneamento básico criam um alerta para seu grande potencial epidêmico. Por isso, as políticas de saúde pública devem priorizar ações informativas e preventivas a fim de evitar surtos e complicações. A atenção primária à saúde é fundamental nesse contexto, justamente pela longitudinalidade e abrangência do cuidado.

Background: Enteroparasitosis are the focus of scientific research worldwide. Urbanorum spp. was recognized as a parasite in Peru in 1994, expanding throughout South America. Reported for the first time in Brazil, state of Maranhão, in 2018. This report presents the second case in the state of Paraná. Case report: Male patient, 56 years old, 75kg, diabetic, inhabitant of São José dos Pinhais, urban area, seeks primary care for pain on bowel movement, tenesmus and abdominal cramps. Denies diarrhea, fever, bloody stools, recent trip. Abdominal examination, blood count and partial urine without changes. Stool parasitology: urbanorum spp. Prescribed Nitazoxanide 500mg 12/12h for 3 days. Patient returns with improvement of symptomatology and parasitological negative control. Conclusion:Currently, the scarcity of prospective studies and meta-analyzes make clinical-epidemiological design and treatment of parasitosis difficult. The spread of the parasite between extremes of the country in a short period of time, coupled with the lack of basic sanitation create a warning for its great epidemic potential. Therefore, public health policies should prioritize informative and preventive actions in order to avoid outbreaks and complications. Primary health care is fundamental in this context, precisely because of the longitudinally and comprehensiveness of care.

Introducción: Las enteroparasitosis el punto de enfoque de investigaciones científicas en todo el mundo. Urbanorum spp fue reconocido cómo parásito en 1994 en el Peru, expandiéndose en América do Sul. Relatado por primera vez en Brasil, Maranhão, 2018. Este informe se encuentra en segundo lugar en el estado de Paraná. Relato del caso: Paciente masculino, 56 años, 75 kg, diabético, habitante de São José dos Pinhais, área urbana. Búsqueda atención primaria por dolor al defecar, tenesmo, y dolor abdominal. Nega diarrea, fiebre, sangre en heces o viaje reciente. Examen físico abdominal, hemograma e tests de orina sin modificaciones. Análisis parasitología: urbanorum spp. Prescripto Nitazoxanide 500mg 12/12h durante 3 días. Paciente volvió con alivio sintomático e materia fecal negativo. Conclusión: En la actualidad la escasez de estudios prospectivos y metanálisis dificultan la delineación clínico-epidemiológica y el tratamiento de la parasitosis. La diseminación del parásito entre los extremos del país en un corto período de tiempo, junto con la falta de saneamiento básico, crea una alerta por su gran potencial epidémico. Por lo tanto, las políticas de salud pública deben priorizar las acciones informativas y preventivas para evitar brotes y complicaciones. La atención primaria de salud es fundamental en este contexto, precisamente por la longitudinalidad y la amplitud de la atención.

Use of Mini-FLOTAC and Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals / Utilização do Mini-FLOTAC e Fill-FLOTAC no diagnóstico rápido de infecções parasitárias de mamíferos em zoológicos

Abstract Animals reared in restricted environments are highly susceptible to gastrointestinal infection by helminths and protozoa and therefore zoos are characterized as being parasite-rich environments. Successful implementation of control programs of these parasites in zoo environment depends upon precise and rapid diagnosing of gastrointestinal infections. The aim of this study was to demonstrate the role of the Mini-FLOTAC technique in combination with Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals. Fecal samples were collected from 70 animals in four different zoos located in central and southern Italy. All the samples were analyzed using Mini-FLOTAC in combination with Fill-FLOTAC. Out of the 70 pooled samples examined, 80% (24/30) were positive for at least one parasite. Among the gastrointestinal nematodes, Strongyles were the most frequent (40%), followed by Trichuris spp. (23.3%), Parascaris spp. (13.3%) and Capillaria spp. (3.3%). Among the protozoa, Blastocystis spp., Giardia spp. and Eimeria spp. were detected in 6.6%, 3.3% and 3.3%, respectively. These results show that Mini-FLOTAC in combination with Fill-FLOTAC can be used, not only for rapidly diagnosing parasitic infections in zoo mammals, but also for monitoring control programs in which large numbers of fecal samples need to be examined rapidly and reliably.

Resumo Animais criados em ambiente restritos são altamente suscetíveis a infecção gastrointestinal por helmintos e protozoários, constituindo os zoológicos em ambientes com alta contaminação por parasitos. O sucesso da implementação de programas de controle contra estes parasitos em zoológicos depende do rápido diagnóstico das infecções por parasitas gastrointestinais. O objetivo deste estudo foi demonstrar o papel da técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC no diagnóstico rápido das infecções parasitárias em mamíferos em zoológicos. Amostras de fezes foram coletadas de 70 animais de quatro diferentes zoológicos no centro e sudoeste da Itália. Todas as amostras foram analisadas pela técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC. Do total de 70 pools de fezes examinadas, 80% (24/30) foram positivas para pelo menos um parasito. Entre os nematoides gastrointestinais a maior frequência foi observada para estrongilídeos (40%), seguida por Trichuris spp. (23,3%), Parascaris spp. (13,3%) e Capillaria spp. (3,3%). Entre os protozoários Blastocystis spp., Giardia spp. e Eimeria spp. foram detectados em 6,6%, 3,3% e 3,3%, respectivamente. Estes resultados demonstram que a técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC pode ser utilizada não somente para o diagnóstico rápido das infecções parasitárias em mamíferos em zoológicos, mas também no monitoramento de programas de controle onde grande número de amostras fecais devem ser examinadas de forma rápida e confiável.

Infección pulmonar por Lophomonas sp en una paciente con leucemia mieloide aguda / Lung infection by lophomonas spp. in a female patient with acute myeloid leukemia

RESUMEN Las infecciones por protozoos son prevalentes a nivel mundial, en particular en pacientes inmunosuprimidos. Comunicamos el caso de una paciente procedente de la ciudad de Viña del Mar, Chile, portadora de leucemia mieloide aguda en quiense confirmó una infección por Lophomonas sp. en lavado bronquioalveolar. Se manejó con antibióticos, pero falleció decomplicaciones de su enfermedad de base. Existe poca literatura disponible respecto a este microorganismo. Concluimos que debe considerarse a Lophomonas sp. como posibilidad diagnóstica si se encuentran protozoos en lavados bronquioalveolares de pacientes inmunosuprimidos.

ABSTRACT Protozoic infections are prevalent worldwide, particularly in immunosuppressed patients. We reported the case of a patient from the city of Viña del Mar, Chile, a carrier of acute myeloid leukemia in whom an infection by Lophomonas sp. was confirmed by bronchoalveolar lavage. She was treated with antibiotics but died of complications of the underlying disease. There is little literature available on this microorganism. We conclude that Lophomonas sp. should be considered as a diagnostic possibility if protozoa are found in bronchoalveolar lavage of immunosuppressed patients.

Detection of natural occurrence of Tritrichomonas foetus in cats in Araçatuba, São Paulo, Brazil / Detecção da ocorrência natural de Tritrichomonas foetus em gatos em Araçatuba, São Paulo

The aim of this study was to investigate the occurrence of Tritrichomonas foetus in cats in the area surrounding the city of Araçatuba municipality, State of São Paulo, Brazil. Fecal samples from 129 cats were collected by rectal flush technique. It was compared two diagnosis methods, direct examination of feces and PCR. The presence of T. foetus DNA was verified using PCR by amplification of 347-bp fragment from the primers TFR3 and TFR4 and amplicons of positive cases were sequenced. Statistical analyses were performed investigating the associations between T. foetus infection with gender, age, breed, presence of diarrhea and/or history of diarrhea, previous treatment, lifestyle, origin, environment, and co-infection. T. foetus was observed in one sample (n=129) by direct microscopic examination of feces while PCR was positive in five samples (3.9%). Giardia species and Cryptosporidium species co-infection was also observed. Statistical analyses showed no significant associations between T. foetus infection and all listed factors, although most positive cats were asymptomatic and lived in multi-cat household. The isolates of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world. So, the occurrence of T. foetus was confirmed in cats in Araçatuba city (Brazil). This parasite must be considered as a differential diagnosis in cats with diarrhea and also in asymptomatic carriers as source of infection in multi-cat environments.(AU)

O objetivo deste estudo foi investigar a ocorrência de Tritrichomonas foetus em gatos na região do município de Araçatuba, SP, Brasil. Foram coletadas amostras fecais de 129 gatos através da técnica de lavado retal. Dois métodos diagnósticos foram comparados, o exame direto das fezes e a PCR. A presença de DNA de T. foetus foi verificada por meio da PCR através da amplificação de 347 pares de bases a partir dos iniciadores específicos TFR3 e TFR4. Posteriormente, os resultados amplificados das amostras positivas foram sequenciadas. Também foi feita análise estatística a fim de investigar a correlação entre infecção por T. foetus e sexo, idade, raça, presença e/ou histórico de diarreia, tratamento prévio, coinfecção, estilo de vida, origem e tipo de ambiente. O protozoário pôde ser observado em uma amostra através do exame direto das fezes e à PCR foram detectadas cinco amostras positivas (3.9%). Foram detectadas coinfecções por Giardia spp. e Cryptosporidium spp. Não foram observadas correlações entre infecção por T. foetus e todos os fatores listados anteriormente, embora a maioria dos felinos positivos fossem assintomáticos e vivessem em ambientes multigatos. O resultado do sequenciamento genético dos isolados das amostras positivas mostrou 100% de similaridade com outros isolados de felinos no mundo. Assim, a ocorrência de T. foetus foi confirmada em gatos em Araçatuba, São Paulo, Brasil. Sendo assim, o parasito deve considerado como diagnóstico diferencial em gatos com diarreia assim como em portadores assintomáticos como fontes de infecção em ambientes multigatos.(AU)

Concordance between the zinc sulphate flotation and centrifugal sedimentation methods for the diagnosis of intestinal parasites / Concordancia entre los métodos de flotación con sulfato de zinc y sedimentación centrífuga para el diagnóstico de parásitos intestinales

Resumen Introduction: The diagnosis of intestinal parasitic infections depends on the parasite load, the specific gravity density of the parasite eggs, oocysts or cysts, and the density and viscosity of flotation or sedimentation medium where faeces are processed. Objective: To evaluate the concordance between zinc sulphate flotation and centrifugal sedimentation in the recovery of parasites in faecal samples of children. Materials and methods: Faecal samples of 330 children from day care centers were evaluated by zinc sulphate flotation and centrifugal sedimentation techniques. The frequencies of detection of parasites by each method were determined and the agreement between the diagnostic techniques was evaluated using the kappa index, with 95% confidence intervals. Results: The faecal flotation in zinc sulphate diagnosed significantly more cases of Trichuris trichiura infection when compared to centrifugal sedimentation (39/330; 11.8% vs. 13/330; 3.9%, p<0.001), with low diagnostic concordance between methods (kappa=0.264; 95% CI: 0.102-0.427). Moreover, all positive samples for Enterobius vermicularis eggs (n=5) and Strongyloides stercoralis larvae (n=3) were diagnosed only by zinc sulphate. No statistical differences were observed between methods for protozoa identification. Conclusions: The results showed that centrifugal flotation in zinc sulphate solution was significantly more likely to detect light helminths eggs such as those of T. trichiura and E. vermicularis in faeces than the centrifugal sedimentation process.

Abstract Introducción. El diagnóstico de infecciones parasitarias intestinales depende de la carga de parásitos, la densidad de la gravedad específica de los huevos, ooquistes o quistes de parásitos, y de la densidad y viscosidad de los reactivos de flotación o sedimentación usados para procesar las heces. Objetivo. Evaluar la concordancia entre el método de flotación de sulfato de zinc y la sedimentación por centrifugación en la recuperación de parásitos en muestras fecales de niños. Materiales y métodos. Se evaluaron las muestras fecales de 330 niños de guarderías mediante las técnicas de flotación con sulfato de zinc y de sedimentación por centrifugación. Se determinó la frecuencia de detección de parásitos con cada método y se evaluó la concordancia entre las técnicas de diagnóstico mediante el índice kappa, con intervalos de confianza del 95 %. Resultados. Mediante la flotación fecal con sulfato de zinc, se diagnosticó un número significativamente mayor de casos de infección por Trichuris trichiura que con la sedimentación por centrifugación (39/330; 11,8 % Vs. 13/330; 3,9 %) (p<0,001), con poco acuerdo entre los métodos (kappa=0,264; IC95% 0,102-0,427). Además, todas las muestras positivas para huevos de Enterobius vermicularis (n=5) y larvas de Strongyloides stercoralis (n=3) se diagnosticaron solamente por sulfato de zinc. No se observaron diferencias estadísticamente significativas entre los métodos para la identificación de protozoos. Conclusiones. La flotación centrífuga en una solución de sulfato de zinc presentó una probabilidad significativamente mayor de detectar los huevos livianos de helmintos como T. trichiura y E. vermicularis en heces, que el proceso de sedimentación por centrifugación.

Perkinsus beihaiensis infecting the oyster Crassostrea rhizophorae under cultivation and in natural stock in Camamu Bay, Bahia, Brazil / Perkinsus beihaiensis infectando a ostra Crassostrea rhizophorae em cultivo e em estoque natural na Baía de Camamu, Bahia, Brasil

This study investigated the infection of Perkinsus beihaiensis (Perkinsozoa) in the oyster Crassostrea rhizophorae, both from a long-line cultivation system and from a nearby intertidal zone of mangrove, both in the state of Bahia, northeastern Brazil. The collections were performed in October and November 2012, and in January 2013. The oysters (n = 300) were measured, examined macroscopically for signs of infection and then submitted to the following laboratory techniques: histology, Ray"s fluid thioglycollate medium assay (RFTM), polymerase chain reaction (PCR) and sequencing, which confirmed the identification of the pathogen. Histological and RFTM analyses showed, respectively, a mean prevalence of 93.3% and of 69%. The infection was usually mild or very mild. There was no significant difference (p > 0.05) between the environments in terms of infection prevalence or severity. This is the first record of P. beihaiensis in the state of Bahia and the second in oysters from Brazil and South America...

Foi investigada a infecção de Perkinsus beihaiensis (Perkinsozoa) na ostra Crassostrea rhizophorae em um sistema de cultivo do tipo espinhel e em um estoque natural de ostras no manguezal adjacente ambos localizados no estado da Bahia, Nordeste do Brasil. As colheitas foram realizadas em outubro e novembro de 2012 e em janeiro de 2013. As ostras (n = 300) foram medidas, examinadas macroscopicamente quanto a sinais da infecção e submetidas às técnicas laboratoriais: histologia, ensaio em meio de cultivo de tioglicolato de Ray (RFTM), reação em cadeia da polimerase (PCR) e sequenciamento, que confirmou a identificação do patógeno. As análises histológicas e o RFTM mostraram, respectivamente, prevalência média de 93,3% e de 69%. A infecção foi geralmente leve ou muito leve. Não houve diferença significativa (p > 0,05) entre os ambientes em termos de prevalência ou severidade da infecção. Este é o primeiro registro de P. beihaiensis no estado da Bahia e o segundo em ostras do Brasil e América do Sul...

Molecular identification of Hepatozoon canis in dogs from Campo Grande, Mato Grosso do Sul, Brazil / Identificação molecular de Hepatozoon canis em cães de Campo Grande, Mato Grosso do Sul, Brasil

The aim of this study was to investigate the occurrence of Hepatozoon species infecting dogs in the municipality of Campo Grande, Mato Grosso do Sul (MS), Brazil, using blood samples (n = 165) drawn from dogs. The species Hepatozoon caniswas identified in 3.63% of the tested animals using molecular tools. Further studies are needed to determine the clinical relevance of this infection and the main arthropod vectors involved in its transmission.

O objetivo deste estudo foi identificar a frequência e espécies de Hepatozoon infectando cães no município de Campo Grande, Mato Grosso do Sul, Brasil. Uma amostragem de 165 animais foi utilizada e, por meio do uso de ferramentas moleculares, a espécie Hepatozoon canis foi identificada em 3,63% dos animais. Mais estudos são necessários para identificar a relevância clínica e os principais vetores envolvidos na transmissão desse protozoário na região.

Prevalence of ittestinal parasites among food handlers in western Iann / La prevalencia de parasitos intestinales entre los manipuladores de alimentos en el oeste de Iran

Parasitic infection is one of the problems that affect human health, especially in developing countries. In this study, all of the fast food shops, restaurants, and roast meat outlets of Khorramabad (Western Iran) and all the staff employed by them, some 210 people, were selected through a census and their stools were examined for the presence of parasites. The parasitological tests of direct wet-mount, Lugol's iodine staining, formaldehyde-ether sedimentation and Trichrome staining techniques were performed on the samples. The data was analyzed with a chi-square test and logistic regression was selected as the analytical model. The results showed 19 (9%) stool specimens were positive for different intestinal parasites. These intestinal parasites included Giardia lamblia 2.9%, Entamoeba coli 4.3%, Blastocystis sp. 1.4%, and Hymenolepis nana 0.5%. There was a significant difference between the presence of a valid health card, awareness of transmission of intestinal parasites, participation in training courses in environmental health with intestinal parasites (p < 0.05). No statistically significant difference was found between the rate of literacy and gender among patients infected with intestinal parasites (p > 0.05). To control parasitic infection in food handlers, several strategies are recommended such as stool examinations every three months, public education, application of health regulations, controlling the validity of health cards and training on parasitic infection transmission. In this regard, the findings of the present study can be used as a basis to develop preventive programs targeting food handlers because the spread of disease via them is a common problem worldwide.

Infección parasitaria es uno de los problemas de salud humana, especialmente en los países en desarrollo. En este estudio, todas las tiendas de comida rápida, restaurantes, y tiendas de carne asada en Khorramabad (oeste de Irán) y todo el personal que trabaja en ellos, incluyendo 210 personas fueron seleccionadas a través de los censos y las heces fueron examinadas para detectar la presencia de parásitos. La prueba parasitológica directa de mojado de montaje, la tinción de Lugol's yodo, sedimentación formaldehído-éter y tricrómicas técnicas de tinción se realizaron sobre las muestras. Los datos fueron analizados con la prueba de chi-cuadrado y regresión logística seleccionado como el modelo analítico. Los resultados mostraron 19 (9%) muestras de heces fueron positivos para diferentes parásitos intestinales. Los parásitos intestinales incluyen Giardia lamblia 2.9%, Entamoeba coli 4.3%, Blastocystis sp. 1.4%, Hymenolepis nana 0.5%. Hubo diferencia significativa entre la presencia de la tarjeta sanitaria válida, el conocimiento de la transmisión de los parásitos intestinales, la participación en cursos de formación en materia de salud ambiental con parásitos intestinales (p < 0.05). No hubo diferencia estadísticamente significativa entre la tasa de alfabetización y el sexo entre los pacientes infectados con parásitos intestinales (p > 0.05). Para controlar la infección parasitaria en los manipuladores de alimentos se recomiendan varias estrategias, tales como examen de heces cada tres meses, la educación pública, se aplican las normas sanitarias, el control de validez de la tarjeta sanitaria y la transmisión formación infección parasitaria. En este sentido, los resultados del presente estudio se pueden utilizar como una base para desarrollar programas de prevención dirigidos a los manipuladores de alimentos debido a la propagación de enfermedades a través de ellos es un problema común en todo el mundo.

Evaluation of a Novel Array-Based Toxoplasma, Rubella, Cytomegalovirus, and Herpes Simplex Virus IgG Enzyme Linked Immunosorbent Assay and Its Comparison with Virion/Serion Enzyme Linked Immunosorbent Assays

BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by kappa-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with kappa-coefficients showing near-perfect agreement for the HSV (kappa=0.87), rubella (kappa=0.92) and CMV (kappa=0.93) and substantial agreement for the toxoplasma (kappa=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.

DNA Extraction from Protozoan Oocysts/Cysts in Feces for Diagnostic PCR

PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. QIAamp(R) DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume (50-100 microl) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, approximately 2 oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.

Prevalence and risk factors for intestinal protozoa infection in elderly residents at Long Term Residency Institutions in Southeastern Brazil / Prevalência e fatores de risco para infecção por protozoários intestinais em idosos residentes nas Instituições de Longa Permanência no Sudeste do Brasil

This study determined the prevalence of intestinal protozoa in Long Term Residency Institutions for the Elderly (ILPI) in elders, nurses and food handlers, identifying the risk factors associated with the infections. Stool samples taken from the elderly (n = 293), nurses (63) and food handlers (19) were studied. Questionnaires were used with questions related to sociodemographic variables, health, behavior and health characteristics. Stool samples were examined using the techniques of Faust and Ziehl Neelsen, and the prevalence of G. duodenalis, Cryptosporidium spp., E. histolytica/dispar in the elderly was 4.0%, 1.0% and 0.3% respectively. Nurses and food handlers showed 4.8% and 5.2% positivity only for G. duodenalis, respectively. The origin of the individuals and contact with domestic animals has been associated with infection by G. duodenalis in the elderly, and contact with domestic animals was considered a risk factor for infection. The last stool examinations were related to Cryptosporidium spp.. None of the variables were associated with E. histolytica/dispar. The frequency of hand washing was significantly associated with G. duodenalis among nurses. The frequency of positive samples of G. duodenalis, Cryptosporidium spp., E. histolytica/dispar showed that ILPIs environments are conducive to this occurring due to contact between the elderly, nurses and food handlers, which are often poorly trained in hygiene procedures and food handling.

Este estudo determinou a prevalência de protozoários intestinais em Instituições de Longa Permanência para Idosos (ILPI), enfermeiros, manipuladores de alimentos identificando fatores de risco associados às infecções. Amostras de fezes de idosos (n = 293), enfermeiros (63), manipuladores de alimentos (19) foram estudadas. Foram aplicados questionários relacionados a variáveis sociodemográficas, de higiene, comportamento e características de saúde. As amostras de fezes foram examinadas pelas técnicas de Faust e Ziehl Neelsen e a prevalência de G. duodenalis, Cryptosporidium spp., E.histolytica/dispar nos idosos foi de 4,0%, 1,0% e 0,3%, respectivamente. Enfermeiros e manipuladores de alimentos apresentaram 4,8% e 5,2% de positividade apenas para G. duodenalis, respectivamente. A origem dos indivíduos, o contato com animais domésticos foram associados à infecção por G. duodenalis nos idosos, sendo que o contato com animais domésticos foi considerado um fator de risco para a infecção, o tempo do último exame coproparasitológico foi relacionado a Cryptosporidium spp.. Nenhuma das variáveis apresentaram associação com E. histolytica/dispar. A frequência da lavagem das mãos foi associada significativamente com G. duodenalis entre enfermeiros. As frequências de amostras positivas para G. duodenalis, Cryptosporidium spp., E. histolytica/dispar demonstram que ILPIs são ambientes propícios para esta ocorrência devido ao contato entre idosos, enfermeiros, manipuladores de alimentos, que muitas vezes são mal treinados em procedimentos de higiene e manipulação de alimentos.

Protozoarios intestinales en escolares adscritos a instituciones públicas y privadas del municipio Maracaibo, estado Zulia / Intestinal protozoa in school children at public and private institutions in the Maracaibo municipality, state of Zulia

Objetivo: Determinar la prevalencia de protozoarios intestinales en escolares adscritos a instituciones públicas y privadas del municipio Maracaibo, Estado Zulia. Materiales y Métodos: Se analizaron 133 muestras fecales de escolares de uno y otro sexo, provenientes de escuelas públicas y unidades educativas privadas con edades comprendidas entre 6 y 14 años. A cada escolar se le analizó una muestra fecal mediante un examen coproparasitológico directo con solución salina fisiológica al 0,85%, coloración temporal de lugol y el método de concentración de Formol-Éter. Resultados: Se observó un predominio de niños parasitados en las instituciones públicas de (39,8%), mientras que en las instituciones privadas fue de (17,3%). El Poliparasitismo en los escolares que asisten a las escuelas públicas fue de (41,3%), superando al resultado obtenido en las instituciones privadas el cual fue de (22,7%). Las especies de protozoarios encontradas fueron: Blastocystis sp que predominó tanto en la población escolar perteneciente a las escuela públicas como en las privadas (50% y 59% respectivamente), Giardia intestinalis (41% en las instituciones privadas y 13% en las públicas). Otros protozoarios observados fueron: Endolimax nana (19%), Entamoeba coli (9%) y Complejo Entamoeba histolytica/Entamoeba dispar (9%), estas últimas especies solo se observaron en las escuelas públicas. Al relacionar las variables parasitosis y grupos etarios estos resultaron estadísticamente significativos, siendo las edades más afectadas los niños de 10 a 14 años. Conclusiones: Los resultados obtenidos, son similares a los encontrados en investigaciones previas en escolares. La alta prevalencia de protozoarios, sugiere un patrón de transmisión directa entre la población estudiada o a través del consumo de agua y/o alimentos contaminados fecalmente. Se plantea la necesidad de evaluar el comportamiento en la higiene alimentaria y abastecimiento de agua.

Objetive: To determine the prevalence of intestinal protozoa in school children at public and private institutions in the Maracaibo Municipality, State of Zulia. Materials and Methods: One-hundred thirty-three (133) fecal samples were analyzed from school children of both sexes, 6 to 14 years old, attending public schools and private educational establishments. For each school child, a stool sample was analyzed by direct coproparasitologic examination using saline 0.85%, temporary lugol coloration and the formol-ether concentration method. Results: A predominance of parasitized children was observed in public institutions (39.8)%, while private institutions evidenced 17.3%. Polyparasitism in children attending public schools was 41.3%, exceeding the result obtained for private institutions, which was 22.7%. Protozoan species found were: Blastocystis sp in both public and private school populations (50% and 59%, respectively); Giardia intestinalis (41% in private institutions and 13% in government schools). Other protozoa observed were: Endolimax nana (19%), Entamoeba coli (9%) and complex Entamoeba histolytica / Entamoeba dispar (9%); the latter species were observed only in public schools. Relating the variables parasitosis and age groups produced statistically significant results; the most affected group of children was from 10 to 14 years old. Conclusions: The results are similar to those obtained from previous research in schools. The high prevalence of protozoa suggests a direct transmission pattern among the studied population either through water consumption and/or fecally contaminated food. This raises the need to evaluate behavior in food hygiene and water supply.

Breast-Feeding Protects Infantile Diarrhea Caused by Intestinal Protozoan Infections

This study investigated the effect of breast-feeding in protection against protozoan infection in infants with persistent diarrhea. Infants were classified into 2 groups; 161 breast-fed infants and the same number of non-breast-fed infants. Microscopic examinations of stool were done for detection of parasites and measuring the intensity of infection. Moreover, serum levels of IgE and TNF-alpha were measured by ELISA. Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Blastocystis sp. were demonstrated in infants with persistent diarrhea. The percentage of protozoan infections was significantly lower in breast-fed infants than that in the non-breast-fed infants. The levels of IgE and TNF-alpha were significantly lower in the breast-fed group than in the non-breast-fed group. There were significant positive associations between the serum levels of IgE and TNF-alpha and the intensity of parasite infection in the breast-fed group. It is suggested that breast-feeding has an attenuating effect on the rate and intensity of parasite infection.

Parasitological stool sample exam by spontaneous sedimentation method using conical tubes: effectiveness, practice, and biosafety / Exame parasitológico de fezes pelo método de sedimentação espontânea utilizando tubos cônicos: eficácia, praticidade e biossegurança

INTRODUCTION: Spontaneous sedimentation is an important procedure for stool examination. A modification of this technique using conical tubes was performed and evaluated. METHODS: Fifty fecal samples were processed in sedimentation glass and in polypropylene conical tubes. Another 50 samples were used for quantitative evaluation of protozoan cysts. RESULTS: Although no significant differences occurred in the frequency of protozoa and helminths detected, significant differences in protozoan cyst counts did occur. CONCLUSIONS: The use of tube predicts a shorter path in the sedimentation of the sample, increases concentration of parasites for microscopy analysis, minimizes the risks of contamination, reduces the odor, and optimizes the workspace.

INTRODUÇÃO: A sedimentação espontânea é um procedimento importante para o exame parasitológico de fezes. Uma modificação desta técnica utilizando tubos cônicos foi realizada e avaliada. MÉTODOS: Cinquenta amostras de fezes foram processadas em cálices de sedimentação de vidro e em tubos cônicos de polipropileno. Outras 50 amostras foram usadas para avaliação quantitativa de cistos de protozoários. RESULTADOS: Embora não tenham sido encontradas diferenças significativas na frequência de helmintos e protozoários identificados, houve diferença significativa na contagem de cistos de protozoários. CONCLUSÕES: O uso do tubo prevê um caminho mais curto na sedimentação da amostra, aumenta a concentração de parasitas para a análise microscópica, minimiza os riscos de contaminação, reduz o odor e otimiza o espaço de trabalho.

A comparative study of the TF-Test®, Kato-Katz, Hoffman-Pons-Janer, Willis and Baermann-Moraes coprologic methods for the detection of human parasitosis

This study compares the diagnostic accuracy of the TF-Test® (TFT) for human parasitosis with results obtained using the traditional Kato-Katz (KK), Hoffman-Pons-Janer (HPJ), Willis and Baermann-Moraes (BM) techniques. Overall, four stool samples were taken from each individual; three alternate-day TFT stool samples and another sample that was collected in a universal container. Stool samples were taken from 331 inhabitants of the community of Quilombola Santa Cruz. The gold standard (GS) for protozoa detection was defined as the combined results for TFT, HPJ and Willis coproscopic techniques; for helminth detection, GS was defined as the combined results for all five coproscopic techniques (TFT, KK, HPJ, Willis and BM). The positivity rate of each method was compared using the McNemar test. While the TFT exhibited similar positivity rates to the GS for Entamoeba histolytica/dispar (82.4 percent) and Giardia duodenalis (90 percent), HPJ and Willis techniques exhibited significantly lower positivity rates for these protozoa. All tests exhibited significantly lower positivity rates compared with GS for the diagnosis of helminths. The KK technique had the highest positivity rate for diagnosing Schistosoma mansoni (74.6 percent), while the TFT had the highest positivity rates for Ascaris lumbricoides (58.1 percent) and hookworm (75 percent); HPJ technique had the highest positivity rate for Strongyloides stercoralis (50 percent). Although a combination of tests is the most accurate method for the diagnosis of enteral parasites, the TFT reliably estimates the prevalence of protozoa and selected helminths, such as A. lumbricoides and hookworm. Further studies are needed to evaluate the detection accuracy of the TFT in samples with varying numbers of parasites.