Prevalence of infection by Cryptosporidium spp. in calves and associated risk factors in Northeastern Brazil / [Prevalência da infecção por Cryptosporidium spp. e fatores de risco associados em bezerros do Nordeste do Brasil]

Cryptosporidium spp. are zoonotic protozoa, frequently associated with diarrhea in calves, which are responsible for important economic losses. The aim of this study was to assess the prevalence of infection by Cryptosporidium spp. and its associated risk factors among calves raised in a milk production region of Northeastern Brazil. Fecal samples (n = 385) were obtained from young animals (up to ten months old) and evaluated by means of centrifugal fecal sedimentation in formalin-ether followed by the modified Ziehl-Neelsen staining technique. In addition, Odds Ratio (OR) was calculated to evaluate associations between variables and infection by these protozoa. Out of all samples analyzed, 25.7% (99/385) scored positive for the presence of Cryptosporidium spp. Contact with other species (goat and sheep) (OR = 3.33; p = 0.000), use of a semi-intensive rearing system (OR = 1.70; p = 0.024) and absence of hygienic conditions (fecal contamination of food and water) (OR = 1.64; p = 0.029) were considered to be risk factors. Data herein reported shows that the implementation of hygienic-sanitary measures on the farms studied, it is imperative to reduce Cryptosporidium spp. infection and consequently the economic impact caused by this pathogen.(AU)

Cryptosporidium spp. são protozoários zoonóticos frequentemente associados à diarreia em bezerros e responsáveis por importantes perdas econômicas. O objetivo deste estudo foi avaliar a prevalência e os fatores de risco associados à infecção por Cryptosporidium spp. em bezerros de propriedades leiteiras no Nordeste do Brasil. Amostras fecais (n = 385) foram obtidas de animais jovens (até 10 meses de idade) e avaliadas por centrífugo-sedimentação em formol éter, seguida da técnica de coloração de Ziehl-Neelsen modificada. A Odds Ratio (OR) foi calculada para avaliar a associação entre variáveis e infecção pelos protozoários. De todas as amostras analisadas, 25,7% (99/385) apresentaram oocistos de Cryptosporidium spp. Contato com outras espécies (caprino e ovino) (OR = 3,33; p = 0,000), sistema semi-intensivo de criação (OR = 1,70; p = 0,024) e ausência de condições higiênicas (contaminação fecal do alimento e da água) (OR = 1,64; p = 0,029) foram considerados fatores de risco. Com base nos resultados, é imprescindível a adoção de medidas higiênico-sanitárias nas fazendas estudadas, a fim de reduzir infecção por Cryptosporidium spp. e o impacto econômico causado por esse patógeno.(AU)

Intestinal coccidian: an overview epidemiologic worldwide and Colombia / Coccidias Intestinales: Panorama epidemiológico mundial y en Colombia

Intestinal coccidia have been classified as protozoa of the Apicomplex phylum, with the presence of an intracellular behavior and adaptation to the habit of the intestinal mucosa, related to several parasites that can cause enteric infections in humans, generating especially complications in immunocompetent patients and opportunistic infections in immunosuppressed patients. Alterations such as HIV/AIDS, cancer and immunosuppression. Cryptosporidium spp., Cyclospora cayetanensis and Cystoisospora belli are frequently found in the species. Multiple cases have been reported in which their parasitic organisms are associated with varying degrees of infections in the host, generally characterized by gastrointestinal clinical manifestations that can be observed with diarrhea, vomiting, abdominal cramps, malaise and severe dehydration. Therefore, in this review a specific study of epidemiology has been conducted in relation to its distribution throughout the world and in Colombia, especially, global and national reports about the association of coccidia informed with HIV/AIDS. Proposed revision considering the needs of a consolidated study in parasitology, establishing clarifications from the transmission mechanisms, global and national epidemiological situation, impact at a clinical level related to immunocompetent and immunocompromised individuals, as well as a focus on public health in institutional government policies and scientific information based on the characterization of coccidia in the tropical region and Colombia.

Los coccidios intestinales se han clasificado como protozoos del Apicomplexa phylum, con presencia de un comportamiento intracelular y adaptación al hábito de la mucosa intestinal, relacionado con varios parásitos que pueden causar infecciones entéricas en los humanos, generando especialmente complicaciones en pacientes inmunocompetentes e infecciones oportunistas en pacientes inmunodeprimidos. Alteraciones como el VIH/SIDA, cáncer e inmunosupresión con tratamientos farmacológicos. En las especies que se encuentran frecuentemente se encuentran Cryptosporidium spp., Cyclospora cayetanensis y Cystoisospora belli. Se han reportado múltiples casos en los que sus organismos parásitos se asocian a diversos grados de infecciones en el huésped, generalmente caracterizadas por manifestaciones clínicas gastrointestinales que pueden observarse con diarrea, vómitos, calambres abdominales, malestar general y deshidratación severa. Por lo tanto, en esta revisión se ha realizado un estudio específico de epidemiología con relación a su distribución en todo el mundo y en Colombia, especialmente, reportes a nivel global y nacional a cerca de la asociación de coccidios informados con el VIH/SIDA. Revisión propuesta con el objetivo de considerar las necesidades de un estudio consolidado a nivel del campo de la parasitología, evidenciando literatura actualizada, estableciendo información de los mecanismos de transmisión, situación epidemiológica global y nacional, impacto a nivel clínico relacionadas con individuos inmunocompetentes e inmunocomprometidos, así como un enfoque en salud pública en políticas gubernamentales institucionales y la información científica basada en la caracterización de coccidias en la región tropical y principalmente en Colombia.

Cryptosporidium spp. e Giardia spp. em carnívoros silvestres do Estado do Pará ­ Brasil / Cryptosporidium spp. and Giardia spp. in wild carnivorous animals the State of Pará ­ Brazil

O presente estudo investigou a ocorrência de Cryptosporidium spp. e Giardia spp. em diferentes espécimes silvestres da ordem Carnívora de vida livre e de cativeiro procedentes de municípios do Estado do Pará. Coletou-se amostras fecais de 37 animais distintos (quatro de vida livre e 33 de cativeiro). Para pesquisa de Cryptosporidium spp. e Giardia spp. foram utilizados métodos microscópicos (direto e Kinyoun) e imunológico (RIDA®QUICK Cryptosporidium/Giardia/Entamoeba Combi - N1722). Do total de amostras, 24,32% (9/37) foram positivas, correspondendo a 5,4% (2/37) para Cryptosporidium spp. e 18,91% (7/37) para Giardia spp., respectivamente. Nenhum animal apresentou infecção concomitante para os agentes. Cryptosporidium spp. e Giardia spp., são protozoários zoonóticos que representam um emergente problema de saúde pública. Esses parasitos podem apresentar elevada frequência em regiões em que as condições de saneamento básico são precárias, promovendo surtos de diarreia em animais domésticos, silvestres e no homen. Mamíferos silvestres, como os carnívoros, são susceptíveis à contaminação por enteroparasitas presentes tanto no habitat natural como em cativeiro. Portanto, a pesquisa comprova a presença desses protozoários em carnívoros silvestres, tanto mantidos em criatórios como nos de vida livre no Estado do Pará, considerando-se que esses animais podem atuar como fontes de infecção para o homem, para outros animais e para o meio ambiente.

The presente survey has had the purpose to investigate the occurrance of Cryptosporidium spp. and Giardia spp. in free and under captivity carnivorous wild animals, from several counties in the State of Pará. Samples of feces from 37 distinct animals (four in their natural habitat and 33 raised in captivity). For the research of Cryptosporidium spp. and Giardia spp. microscopic immunological, direct and Kinyoun methods were used (RIDA®QUICK Cryptosporidium/Giardia/Entamoeba Combi - N1722). The samples gathered from wild animals have resulted in 24,32% of positive infecction on the rate of (9/37), being. 5,4% (2/37) positive to Cryptosporidiumspp. and 18,91% (7/37) positive to Giardia spp., what shows that no amimals had both infections at the same time. Cryptosporidium spp. and Giardia spp., are zoonotic enteroparasites that have been taking place as an emmerging problem to public health. Theese species of protozoa may reach high levels of frequency in regions where the basic sanitation conditions are precarious, promoting outbraks of diarrhea to men, wild and domestic animals. Wild mammals, as the carnivorous, are susceptible to contamination by enteroparasites, being present at their natural habitat or captivity. So, the reserach strenghtens the real presence of these protozoas in wild carnivorous in both conditions of life, free or under captivity, in the State of Pará, making us consider the possibility that the cited animals may be natural reservoirs for infections, not only to men but to other animals and also to environment.

Cryptosporidium spp. e Giardia spp. em carnívoros silvestres do Estado do Pará Brasil / Cryptosporidium spp. and Giardia spp. in wild carnivorous animalsthe State of Pará – Brazil

O presente estudo investigou a ocorrência de Cryptosporidium spp. e Giardia spp. em diferentes espécimes silvestres da ordem Carnívora de vida livre e de cativeiro procedentes de municípios do Estado do Pará. Coletou-se amostras fecais de 37 animais distintos (quatro de vida livre e 33 de cativeiro). Para pesquisa de Cryptosporidium spp. e Giardia spp. foram utilizados métodos microscópicos (direto e Kinyoun) e imunológico (RIDA®QUICK Cryptosporidium/Giardia/Entamoeba Combi - N1722). Do total de amostras, 24,32% (9/37) foram positivas, correspondendo a 5,4% (2/37) para Cryptosporidium spp. e 18,91% (7/37) para Giardia spp., respectivamente. Nenhum animal apresentou infecção concomitante para os agentes. Cryptosporidium spp. e Giardia spp., são protozoários zoonóticos que representam um emergente problema de saúde pública. Esses parasitos podem apresentar elevada frequência em regiões em que as condições de saneamento básico são precárias, promovendo surtos de diarreia em animais domésticos, silvestres e no homen. Mamíferos silvestres, como os carnívoros, são susceptíveis à contaminação por enteroparasitas presentes tanto no habitat natural como em cativeiro. Portanto, a pesquisa comprova a presença desses protozoários em carnívoros silvestres, tanto mantidos em criatórios como nos de vida livre no Estado do Pará, considerando-se que esses animais podem atuar como fontes de infecção para o homem, para outros animais e para o meio ambiente.

The present survey has had the purpose to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in free and under captivity carnivorous wild animals, from several counties in the State of Pará. Samples of feces from 37 distinct animals (four in their natural habitat and 33 raised in captivity). For the research of Cryptosporidium spp. and Giardia spp. microscopic immunological, direct and Kinyoun methods were used (RIDA®QUICK Cryptosporidium/Giardia/Entamoeba Combi - N1722). The samples gathered from wild animals have resulted in 24,32% of positive infection on the rate of (9/37), being. 5,4% (2/37) positive to Cryptosporidium spp. and 18,91% (7/37) positive to Giardia spp., what shows that no animals had both infections at the same time. Cryptosporidium spp. and Giardia spp., are zoonotic enteroparasites that have been taking place as an emerging problem to public health. These species of protozoa may reach high levels of frequency in regions where the basic sanitation conditions are precarious, promoting outbreaks of diarrhea to men, wild and domestic animals. Wild mammals, as the carnivorous, are susceptible to contamination by enteroparasites, being present at their natural habitat or captivity. So, the research strengthens the real presence of these protozoas in wild carnivorous in both conditions of life, free or under captivity, in the State of Pará, making us consider the possibility that the cited animals may be natural reservoirs for infections, not only to men but to other animals and also to environment.

Molecular Prevalence of Cryptosporidium spp. in Breeding Kennel Dogs

Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99–100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.

The effect of water source and soil supplementation on parasite contamination in organic vegetable gardens / O efeito da fonte de água e suplementação de solo na contaminação parasitária em hortas orgânicas

Abstract The objective of this study was to determine factors associated with vegetable contamination with zoonotic protozoan. Samples of water, soil and vegetables were collected from July/2014 to May/2016, totaling 83 samples, 21 properties of Londrina region, Paraná, Brazil. DNA amplification of Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis in the samples was conducted using polymerase chain reaction (PCR). The PCR results were positive for T. gondii in 12.9% (8/62), Cryptosporidium spp. in 11.3% (7/62) and G. intestinalis in 25.8% (16/62) of the samples. DNA sequencing identified C. parvum in five samples and G. intestinalis Assemblage E in three. The statistical associations demonstrated greater probability of positive samples for T. gondii and for at least one of the three protozoa when the source of irrigation water was the river; a greater chance of positive samples for Cryptosporidium spp. when deer were present on the property; and a smaller chance of positive samples for at least one of the three etiologic agents when soil was supplemented with limestone. The results expose some critical contamination points, providing support for training farmers on good management practices during the production process.

Resumo O trabalho teve como objetivo determinar os fatores associados à contaminação de vegetais por protozoários zoonóticos. Amostras de água, solo e vegetais foram coletadas de julho/2014 a maio/2016, totalizando 83 amostras de 21 propriedades da região de Londrina, Paraná, Brasil. A amplificação de fragmentos de DNA de T. gondii, Cryptosporidium spp. e Giardia intestinalis foi realizada por meio da reação em cadeia da polimerase (PCR). Os resultados da PCR foram positivos para T. gondii em 12,9% (8/62), Cryptosporidium spp. em 11,3% (7/62) e G. intestinalis. em 25,8% (16/62) das amostras. O sequenciamento de DNA identificou C. parvum em cinco amostras e G. intestinalis, Assemblage E em três amostras. As associações estatísticas evidenciaram maior probabilidade de amostras serem positivas para T. gondii ou para pelo menos um dos três protozoários quando a fonte de água de irrigação era o rio; uma maior chance de amostras positivas para Cryptosporidium spp. quando havia cervos na propriedade; e uma menor chance das amostras serem positivas para pelo menos um dos três agentes etiológicos quando o solo era suplementado com calcário. Os resultados expõem alguns pontos críticos de contaminação, fornecendo suporte para capacitar os agricultores em boas práticas de gestão durante o processo de produção.

Sensitivity, specificity and comparison of three commercially available immunological tests in the diagnosis of Cryptosporidium species in animals

ABSTRACT The study was conducted to compare the specificity of immunological diagnostic methods used for the diagnosis of Cryptosporidium species capable of causing life-threatening infection in both immunosuppressed and immunocompetent patients. For the detection of Cryptosporidium species in 79 animals with diarrhoea, we used three Copro-antigen tests: RIDASCREEN® Cryptosporidium test, Cryptosporidium 2nd Generation (ELISA) and RIDA®QUICK Cryptosporidium. For immunoassays we used positive and negative samples detected by means of polymerase chain reaction and validated by sequencing and nested polymerase chain reaction to confirm the presence six different species of Cryptosporidium species. Prevalence of cryptosporidiosis in the entire group determined by enzyme immunoassay, enzyme linked immunosorbent assay, immuno-chromatographic test and polymerase chain reaction was 34.17%, 27.84%, 6.33% and 27.84%, respectively. Sensitivity of animal samples with enzyme immunoassay, enzyme linked immunosorbent assay, and immuno-chromatographic test was 63.6%, 40.9% and 22.7%, resp., when questionable samples were considered positive, whereas specificity of enzyme immunoassay, enzyme linked immunosorbent assay and immuno-chromatographic test was 75.9%, 78.9% and 100%, respectively. Positive predictive values and negative predictive values were different for all the tests. These differences results are controversial and therefore reliability and reproducibility of immunoassays as the only diagnostic method is questionable. The use of various Cryptosporidium species in diagnosis based on immunological testing and different results obtained by individual tests indicate potential differences in Copro-antigens produced by individual Cryptosporidium species.

Gastrointestinal protozoa in dairy calves: identification of risk factors for infection / Protozoos gastrointestinales en terneros lecheros: identificación de factores de riesgo para la infección

ABSTRACT Objective. This study aimed to evaluate the occurrence of gastrointestinal protozoa in dairy calves and to identify potential risk factors for this type of infection. Materials and methods. For this purpose, 243 fecal samples were collected from calves up to 60 days of age in 43 dairy farms located in the West region of Santa Catarina state, Brazil. Samples were examined by centrifugal-flotation technique. Results. As a result, Giardia was present in 26.75% (65/243) of all samples, Eimeria in 21.81% (53/243), and Cryptosporidium in 20.99% (51/243). Additionally, 46.50% (113/243) of the samples were negative for any protozoa, while 39.10% (95/243) and 14.40% (35/208) showed single and mixed infections, respectively. There was a higher association between Cryptosporidium and Giardia (6.99%) in cases of mixed infections. However, the triple protozoa association had the lowest prevalence in mixed infections (2.06%). Epidemiologically, a questionnaire was applied to determine risk factors for these parasitic infections. Based on the statistical model applied, some risk factors for Cryptosporidium infections were identified, highlighting feeding management, period of time that calves stayed with their mothers (cows), and contact with dogs; the risk of contracting Giardia increased according to the milk source, while the floor type bedding, and age were appointed as risks factors for Eimeria. Conclusions. Therefore, it is possible to confirm that Giardia, Cryptosporidium and Eimeria may infect dairy calves, and the knowledge of some risk factors associated to their infection in calves.

RESUMEN Objetivo. El objetivo evaluar la ocurrencia de protozoos gastrointestinales en terneros lecheros y de identificar posibles factores de riesgo para infección. Materiales y métodos. Se recogieron 243 muestras de heces de terneros de hasta 60 días de edad en 43 granjas lecheras ubicadas en la región del oeste del estado de Santa Catarina, Brasil. Las muestras fueron examinadas por la técnica de centrifugación-flotación. Resultados. Como resultado, Giardia estaba presente en 26.75% (65/243) de todas las muestras, Eimeria en 21.81% (53/243), y Cryptosporidium en 20.99% (51/243). Además, 46.50% (113/243) de las muestras fueron negativas para cualquier protozoos, mientras que 39.10% (95/243) y 14.40% (35/208) mostraron infecciones simples y mixtas, respectivamente. Hubo una mayor asociación entre Cryptosporidium y Giardia (6.99%) en los casos de infecciones mixtas. Sin embargo, la asociación de triple protozoos tenía la menor prevalencia de infecciones mixtas (2.06%). Epidemiológicamente, se aplicó un cuestionario para determinar los factores de riesgo para estas infecciones parasitarias. Se han identificado algunos factores de riesgo de infecciones por Cryptosporidium, destacando manejo de la alimentación, periodo de tiempo que los terneros permanecieron con sus madres (vacas), y el contacto con los perros; el riesgo de contraer Giardia incrementado en función de la fuente de la leche, mientras que el tipo de suelo de las camas, y la edad fueron nombrados como los riesgos de factores de Eimeria. Conclusiones. Es posible confirmar que Giardia, Cryptosporidium y Eimeria pueden infectar a terneros lecheros, y el conocimiento de algunos factores de riesgo asociados a la infección en terneros.

Pesquisa de bioagentes na água do Rio Pardo, Brasil, e estimativa de risco de infecção e de doença por Cryptosporidium spp. e Giardia spp / Research on bioagents in the Pardo river water, Brazil, and estimated risk of infection and disease by Cryptosporidium spp. and Giardia spp

O lançamento de esgotos domésticos in natura, efluentes das estações de tratamento de esgoto e escoamento superficial, são relatados como importantes causas de poluição das águas superficiais. Sabe-se que a alteração da qualidade das águas dos rios restringe seus múltiplos usos e contribui para o aumento de doenças de veiculação hídrica, em decorrência da exposição oral a bioagentes patogênicos. Neste contexto, o objetivo do presente estudo foi identificar e quantificar bioagentes presentes na água do rio Pardo, Brasil, e estimar o risco de infecção e de doença por Cryptosporidium spp. e Giardia spp. para a população, devido ao uso do rio como fonte de abastecimento público e recreação de contato primário, por meio da abordagem da Avaliação Quantitativa de Risco Microbiológico (AQRM). Durante os anos de 2015 e 2016, foram realizadas seis coletas de amostras da água do rio Pardo (período chuvoso e período seco) em seis pontos, totalizando 36 amostras. Foram realizadas análises de identificação e quantificação de E. coli, Salmonella Não Tifóide, Cryptosporidium spp. e Giardia spp. Para estimativa de risco de infecção e de doença por Cryptosporidium spp. e Giardia spp. (AQRM), foram considerados diferentes populações (crianças e adultos), volumes de água ingerido, concentração de (oo)cistos e duração e frequência da exposição, de acordo com o cenário estabelecido. Os valores médios para E. coli variaram de 6,57 x 101 UFC/100 mL a 6,07 x 103 UFC/100 mL, apresentando diferenças estatisticamente significantes (p < 0,05) entre os períodos chuvoso e seco. As densidades de Salmonella Não Tifóide foram baixas (<0,6473 a 1,55 NMP/100 mL), com frequência de 13,9% das amostras positivas, evidenciando a circulação desse patógeno no ambiente. A concentração de (oo)cistos de Cryptosporidium spp. e Giardia spp. variou de <0,1 a 0,4 oocistos/L e <0,1 a 4,4 cistos/L, respectivamente. Para abordagem da AQRM devido a ingestão da água do rio Pardo usada para abastecimento público, a probabilidade anual de infecção por Cryptosporidium spp. e Giardia spp. foi maior para adultos do que para crianças, sendo que na maioria dos pontos apresentou resultados superiores ao risco anual tolerável pela USEPA (1 x 10-4). No que diz respeito ao uso da água do rio Pardo para recreação de contato primário, a probabilidade diária e anual de infecção, bem como a probabilidade de doenças, foi maior para crianças, seguida de adultos/homens e adultos/mulheres. A probabilidade de criptosporidiose e giardíase esteve abaixo do limite tolerável pela USEPA (3,6 x 10-2), exceto no Ponto 4, em que a estimativa de risco de doença por Giardia spp. para crianças esteve acima deste valor. A presença de bioagentes em amostras de água do rio Pardo pode estar relacionada à poluição das águas por fontes pontuais e difusas. Esses achados refletem a importância de priorizar os recursos para implantação e complementação das Estações de Tratamento de Esgoto na UGRHI 4, a fim de prevenir as doenças de veiculação hídrica em populações que utilizam a água do rio Pardo para abastecimento público e recreação de contato primário

The discharge of domestic sewage, effluents of wastewater treatment plants and surface runoff, are reported as important causes of surface water pollution. It is known that the alteration of river water quality restricts its multiple uses and contributes to the increase of waterborne diseases, due to oral exposure to pathogenic bioagents. In this context, the aim of the present study was to identify and quantify bioagents present in Pardo river water, Brazil, and to estimate the risk of infection and disease by Cryptosporidium spp. and Giardia spp. for the population, due to the use of the river as source of public supply and primary contact recreation, through the approach of Quantitative Microbial Risk Assessment (QMRA). During the years of 2015 and 2016, six samples of water from the Pardo river (rainy and dry season) were collected at six points, totaling 36 samples. Identification and quantification analyzes of E. coli, Non-typhoid Salmonella, Cryptosporidium spp. and Giardia spp. To estimate the risk of infection and disease by Cryptosporidium spp. and Giardia spp. (QMRA), different populations (children and adults), volumes of ingested water, concentration of (oo) cysts, duration and frequency of exposure were considered according to the established scenario. Mean values for E. coli varied from 6.57 x 101 CFU / 100 mL to 6.07 x 103 CFU / 100 mL, showing statistically significant differences (p <0.05) between the rainy and dry season. Non-typhoid Salmonella densities were low (<0.6473 at 1.55 MPN / 100 mL), with a frequency of 13.9% of the positive samples, evidencing the circulation of this pathogen in the environment. Cryptosporidium spp. and Giardia spp. concentration ranged from <0.1 to 0.4 oocysts / L and <0.1 to 4.4 cysts / L, respectively. In order to approach the QMRA due to the ingestion of Pardo river water used for public supply, the probability of annual infection by Cryptosporidium spp. and Giardia spp. was higher for adults than for children, and in most points presented results higher than the risk tolerable by USEPA (1 x 10-4). Regarding the use of Pardo river water for primary contact recreation, the daily and annual probability of infection, as well as the probability of illness, was higher for children, followed by adults / men and adults / women. The probability of cryptosporidiosis and giardiasis was below the limit tolerable by USEPA (3.6 x 10-2), except in Point 4, where the estimated risk of disease by Giardia spp. for children was above this value. The presence of bioagents in Pardo river water may be related to water pollution by point and diffuse sources. These findings reflect the importance of prioritizing the resources for implementation and complementation of wastewater treatment plants at UGRHI 4, in order to prevent waterborne diseases in populations that use Pardo river water for public supply and primary contact recreation

Secondary transmission of cryptosporidiosis associated with well water consumption: two case studies

Abstract: Cryptosporidiosis is a very prominent disease in the field of public health, and usually causes diarrhea. We describe two immunocompetent patients who presented with chronic diarrhea that was ultimately found to be caused by continuous exposure to well water contaminated with the microbial cysts (oocysts) of the Cryptosporidium spp parasite. We describe the patients' histories and possible explanations for their prolonged symptoms.

Cryptosporidium infections in birds - a review / Infecção por Cryptosporidium em aves - uma revisão

Cryptosporidiosis is one of the main protozoan infections in birds. It manifests as either a respiratory or a digestive illness, and it affects a very large number of avian species across several continents. The aim of this review is to report on the main results of studies on cryptosporidiosis among birds and the importance of these results to veterinary medicine and public health.

A criptosporidiose constitui-se em uma das principais infecções por protozoários em aves, manifestando-se como enfermidade respiratória ou digestiva, em dezenas de espécies aviárias, em vários continentes. O objetivo desse trabalho foi relatar, por meio de revisão de literatura, os principais resultados de estudos sobre criptosporidiose em aves e sua importância para a medicina veterinária e saúde pública.

Cryptosporidium spp. in bursa of Fabricius of broiler chickens from Uruguay / Cryptosporidium spp. em bursa de Fabrícius de frangos de corte no Uruguai

The presence of Cryptosporidium spp. was detected in broiler chickens of 32-40 days of age, colonizing the bursa of Fabricius. The histologic study with haematoxylin and eosin staining was done on specimens of trachea, complete intestinal tract and bursa. Samples of intestinal content were also studied using Kinyoun staining technique. All birds with signs of illness were positive to the presence of parasite, showing a hypertrophic bursa of Fabricius, filled with caseous content. The bursal epithelial cells were full of different developmental stages of Cryptosporidium spp. This note describing the presence of Cryptosporidium spp. in industrial flocks of broiler chickens, is the first report of this parasite in Uruguay.

A presença de Cryptosporidium spp. foi detectada em frangos de corte de 32-40 dias de idade, colonizando a bursa de Fabrícius. O estudo histológico com hematoxilina e eosina foi feito em amostras de traqueia, trato intestinal completo e bursa. As amostras de conteúdo intestinal também foram estudadas utilizando a técnica de coloração de Kinyoun. Todas as aves com sinais de doença foram positivas para a presença do parasita, mostrando uma bursa de Fabrícius hipertrófica com grande quantidade de conteúdo caseoso. As células epiteliais bursais estavam repletas de diferentes estágios de desenvolvimento de Cryptosporidium spp. Esta nota, que descreve a presença de Cryptosporidium spp. em lotes industriais de frangos de corte, é o primeiro relato deste parasita no Uruguai.

Cryptosporidiosis among children with diarrhoea in three Asian countries:A review

This review focuses on studies concerning cryptosporidiosis in three Asian countries. Cryptosporidium spp. infection was investigated in children<12 years old afflicted with diarrhoea and admitted to the paediatric hospitals in Iraq, Jordan and Malaysia. Most of the patients complained of abdominal pain, watery diarrhoea and mild-to-severe dehydration. Stool samples were collected from children and five methods were used to detect oocysts of Cryptosporidium spp. including:direct wet mount, Sheather’s sugar flotation, formalin-ether sedimentation, modified Ziehl-Neelsen and direct fluorescent antibody (DFA). The infection rate was 8.56, 37.3 and 4.6 in Iraq, Jordan and Malaysia respectively. A combination of formalin ether sedimentation and acid fast stain was used to detect Cryptosporidium oocysts in Iraq. The DFA test showed the highest sensitivity for samples of children in Jordan. In Malaysia, direct wet mount, formalin-ether sedimentation, modified Ziehl-Neelsen and DFA gave the same results (4.62%) while Sheather’s sugar flotation was 3.85%. Source of drinking water appeared to be an important risk factor in transmission of infection. In Jordan, the high rate of infection was recorded in rainy season (January–May).

Cryptosporidiosis among children with diarrhoea in three Asian countries: A review

This review focuses on studies concerning cryptosporidiosis in three Asian countries. Cryptosporidium spp. infection was investigated in children < 12 years old afflicted with diarrhoea and admitted to the paediatric hospitals in Iraq, Jordan and Malaysia. Most of the patients complained of abdominal pain, watery diarrhoea and mild-to-severe dehydration. Stool samples were collected from children and five methods were used to detect oocysts of Cryptosporidium spp. including: direct wet mount, Sheather's sugar flotation, formalin-ether sedimentation, modified Ziehl-Neelsen and direct fluorescent antibody (DFA). The infection rate was 8.56, 37.3 and 4.6 in Iraq, Jordan and Malaysia, respectively. A combination of formalin ether sedimentation and acid fast stain was used to detect Cryptosporidium oocysts in Iraq. The DFA test showed the highest sensitivity for samples of children in Jordan. In Malaysia, direct wet mount, formalin-ether sedimentation, modified Ziehl-Neelsen and DFA gave the same results (4.62%) while Sheather's sugar flotation was 3.85%. Source of drinking water appeared to be an important risk factor in transmission of infection. In Jordan, the high rate of infection was recorded in rainy season (January-May).

Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi in Captive Non-Human Primates in Qinling Mountains

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.

Ocorrência da infecção por Cryptosporidium spp. em cabritos (Capra hircus) / Occurrence of infection by Cryptosporidium spp. in goat kids (Capra hircus)

O presente estudo teve como objetivo determinar a ocorrência da infecção por Cryptosporidium spp. em cabritos de Quixadá, Ceará, Brasil. Participaram do estudo 400 cabritos, com idade entre três e 360 dias, de ambos os sexos, com e sem padrão racial definido, procedentes de 25 estabelecimentos rurais distribuídos em três circuitos. As fezes foram cadastradas de acordo com o aspecto e cor, distribuídas em tubos tipo "eppendorf®" e congeladas in natura a -20°C, até o momento das extrações de DNA genômico do parasito com auxílio de kit comercial. Para amplificação de fragmentos da subunidade 18S do RNA ribossômico (rRNA) foi utilizada a "Nested"-PCR. A ocorrência de Cryptosporidium spp em cabritos de Quixadá foi de 7,50% (30/400). A frequência no período seco e no chuvoso foi de 9,55% (19/199) e 5,47% (11/201), respectivamente (χ²=2,39 e P>0,05). Amostras positivas foram identificadas em 64,00% (16/25) das propriedades estudadas e dessas amostras 50,00% (15/30) e 70,00% (21/30) tinham as fezes com aspecto e cor normais, respectivamente, sugerindo que cabritos assintomáticos estão eliminando oocistos. Não foi observada positividade para Cryptosporidium spp. em animais com 301 a 360 dias, demonstrando que animais mais velhos apresentam menos possibilidade de se infectarem com o parasito...

The present study aimed to determine the occurrence of infection by Cryptosporidium spp. in goat kids from Quixadá, Ceará, Brazil. The study included 400 goat kids of both sexes, 3 to 360 days old, with or without defined breed, originating from 25 farms distributed in three circuits. Feces were registered in accordance with the appearance and color, distributed into tubes Eppendorf tubes and frozen in natura at-20°C until the moment of extraction of genomic DNA from the parasite with the aid of a commercial kit. For amplification of fragments of the 18S subunit of ribosomal RNA (rRNA) was used to Nested PCR. The occurrence of Cryptosporidium spp. in goats kids of the Quixadá was 7.50% (30/400). The frequency in the dry period and rainy was 9.55% (19/199) and 5.47% (11/201) respectively (χ²=2.39 and P>0.05). Positive samples were identified in 64.00% (16/25) of the studied farms, and from these samples 50.00% (15/30) and 70.00% (21/30) had feces with normal appearance and color respectively, suggesting that the asymptomatic goats were eliminating oocysts. No positivity for Cryptosporidium spp. was observed in 301 to 360-day-old goats, demonstrating that older animals have less chance to become infected with the parasite...

Surto de criptosporidiose em bezerros no Sul do Rio Grande do Sul / Outbreak of cryptosporidiosis in calves in Southern Brazil

Descrevem-se os aspectos epidemiológicos, sinais clínicos e a patologia de um surto de criptosporidiose em bezerros na região Sul do Rio Grande do Sul. De um lote de 400 bezerros de 30-45 dias de idade, 35 adoeceram e 16 morreram. Os bezerros nasciam fracos e logo após o nascimento apresentavam diarreia amarela, emagrecimento progressivo, desidratação, depressão e morte entre 10 e 15 dias após o início dos sinais clínicos. Na necropsia havia congestão dos vasos sanguíneos intestinais e mesentéricos. Havia distensão intestinal por gás e dilatação de vasos linfáticos. Microscopicamente havia achatamento das vilosidades intestinais, com necrose e atrofia. Aderidas à superfície das células epiteliais das vilosidades, havia estruturas puntiformes basofílicas de 2-5µm de diâmetro compatíveis com Cryptosporidium spp. A microscopia eletrônica revelou a presença de diferentes estágios do agente aderidos às microvilosidades de enterócitos. Alerta-se para a importância da criptosporidiose como agente primário de diarreia em bezerros. São necessárias medidas preventivas no que se refere ao manejo para diminuir as perdas econômicas e a contaminação ambiental, e, ainda, diminuir o risco para a saúde pública...

This paper describes the epidemiology, clinical signs and pathology of an outbreak of cryptosporidiosis in calves in Southern Brazil. Thirty-five out of 400 calves with 30-45 days of age were affected and 16 died. The calves were born weak and just after birth they had yellow diarrhea, weight loss, dehydration, depression, and death between 10 and 15 days after onset of clinical signs. Congestion of the bowel and mesenteric blood vessels were observed at necropsy. Intestinal distension by gas and dilation of lymphatic vessels were also observed. Microscopically, the intestine showed flattening of the villi with necrosis and atrophy. Adhered to the surface of the villus epithelial cells there were round basophilic structures of 2- to 5-μm-diameter compatible with Cryptosporidium spp. Electron microscopy revealed the presence of different stages of the agent adhered to the microvilli of enterocytes. We alert the importance of cryptosporidiosis as a primary agent of diarrhea in calves. Preventive measures to reduce economic losses, environmental contamination, and also decrease of risk for public health are necessary...

Cryptosporidium spp., Giardia lamblia y encephalitozoon intestinalis, oportunistas emergentes / Cryptosporidium spp., Giardia lamblia y encephalitozoon intestinalis emergent opportunists

Es un hecho conocido que las infecciones oportunistas por protozoos y hongos han aumentado en los últimos años, debido especialmente al aumento de las infecciones por VIH. Cryptosporidium spp., Giardia lamblia y Encephalitozoon intestinalis son protozoos y hongo, respectivamente, mundialmente reconocidos como agentes oportunistas emergentes, responsables de brotes epidémicos provocados por la ingestión de agua potable contaminada, incluso después de una correcta desinfección. La ingestión de estos protozoos puede provocar diferentes grados de enfermedad, entre aguda o leve (población sana) hasta situaciones más graves y agresivas, hasta a veces mortales (pacientes inmunocomprometidos y/o inmunodeprimidos). A pesar de ser responsables de muchos brotes epidémicos, su diagnóstico de laboratorio permanece arduo y trabajoso, incluso utilizando las nuevas técnicas desarrolladas en los últimos años. En esta revisión se resumen las consideraciones generales de estos oportunistas emergentes, así como los métodos de diagnóstico más usuales, incluso los más recientes y específicos.

Epidemiological data, regarding parasitic and fungi opportunist infections, have changed in the last years, especially due to HIV infection. Cryptosporidium spp., Giardia lamblia and Encephalitozoon intestinalis are protozoan and fungi, respectively, worldwide known as opportunistic emergent agents, being responsible by epidemic outbreaks after ingestion of contaminated water, even following a correct disinfection treatment. Its ingestion can cause different effects on individuals’ health, from light or acute among the healthy population, to serious, aggressive or even deadly among the immunodepressed or immunocompromised patients. Contaminated water ingestion can result in outbreaks but protozoa laboratory diagnosis still remains very laborious, even after the development of more sensitive and specific techniques in the last years. In this paper, a revision of these emergent opportunists, their main characteristics and diagnostic tools are described, including the most recent and specific techniques.

Prevalence and genetic characterization of Cryptosporidium spp. and Cystoisospora belli in HIV-infected patients / Prevalência e caracterização genética de Cryptosporidium spp. e Cystoisospora belli em pacientes infectados pelo HIV

Cryptosporidium spp. and Cystoisospora belli are monoxenic protozoa that have been recognized as the causative agents of chronic diarrhea in immunocompromised individuals, especially HIV-infected subjects. The objective of this study was to evaluate the frequency of these intestinal protozoa in HIV-positive patients in the Triângulo Mineiro region of Brazil and to correlate the presence of these infections with clinical, epidemiological and laboratory data of the patients. Oocysts were detected in stool samples of 10 (16.9%) of the 59 patients studied, while Cryptosporidium spp. were present in 10.1% (6/59) and C. belli in 6.7% (4/59). The frequency of these parasites was higher among patients with diarrheic syndrome and CD4+ T lymphocyte counts < 200 cells/mm 3 , demonstrating the opportunistic characteristic of these infections. A significant association was observed between the lack of adherence to antiretroviral therapy and the presence of Cryptosporidium spp. and/or C. belli. Parasitism with Cryptosporidium spp. was more frequent in February and April, the months following the period of high rainfall. The same was not observed for C. belli. Genetic characterization of two isolates led to the identification of Cryptosporidium parvum, one of the main species associated with the zoonotic transmission of cryptosporidiosis.

Cryptosporidium spp. e Cystoisospora belli são protozoários monoxenos reconhecidos como agentes causadores de diarréia crônica em indivíduos imunocomprometidos, especialmente aqueles infectados pelo HIV. Os objetivos deste estudo foram o de avaliar a frequência destes protozoários em pacientes HIV - positivos na região do Triângulo Mineiro, Brasil, e correlacionar a presença destas infecções com dados clínicos, epidemiológicos e laboratoriais dos pacientes. Oocistos foram detectados em amostras fecais de 10 (16,9%) dos 59 pacientes estudados, sendo 10.1% (6/59) das amostras positivas para Cryptosporidium spp. e 6,7% (4/59) das amostras positivas para C. belli. A frequência destes parasitos foi maior entre pacientes com síndrome diarreica e contagem de linfócitos T CD4+ < 200 cells/mm 3 , o que demonstra o caráter oportunista destas infecções. Foi observada uma associação significativa entre a falta de aderência à terapia antiretroviral e a presença de Cryptosporidium spp. e/ou C. belli. Parasitismo por Cryptosporidium spp. foi mais frequente em fevereiro e abril, meses subsequentes ao período chuvoso. O mesmo não foi observado para C. belli. A caracterização genética de dois isolados levou à identificação de Cryptosporidium parvum, uma das principais espécies associadas com a transmissão zoonótica da criptosporidiose.

Identification of Cryptosporidium species and genotypes in dairy cattle in Brazil / Identificação de espécies e genótipos de Cryptosporidium em bovinos leiteiros no Brasil

In this study, we identified Cryptosporidium species and genotypes present in dairy cattle in the central region of São Paulo state, Brazil. Fecal specimens were collected from 200 animals (100 calves and 100 cows) in ten dairy farms. Fecal samples were examined using microscopic examination (ME), enzyme immunoassay (EIA) and polymerase chain reaction (PCR). Cryptosporidium species and genotypes were determined by restriction fragment length polymorphism (RFLP) or DNA sequencing analysis of the SSU-rRNA and GP60 genes. The occurrence of Cryptosporidium spp. infection was 14% (28/200). The occurrence in calves (26%) was significantly higher than in cows (2%). Of the 27 Cryptosporidium-positive specimens submitted to genotyping, C. andersoni was identified in 23 (85.1%), C. bovis in three (11.1%), and the zoonotic C. parvum subtype IIaA15G2R1 in one (3.7%). The study demonstrates that Cryptosporidium spp. infection was common and widespread in dairy cattle in this region and that calves have a high prevalence of C. andersoni. Furthermore, the presence of C. parvum subtype IIaA15G2R1 indicates that dairy calves from this region should be considered a potential source of zoonotic Cryptosporidium oocysts.

No presente estudo foram identificadas espécies e genótipos de Cryptosporidium originadas de bovinos leiteiros na região central do estado de São Paulo, Brasil. Amostras fecais foram coletadas de 200 animais (100 bezerros e 100 vacas) em 10 propriedades leiteiras. As amostras foram examinadas utilizando os métodos de microscopia óptica (MO), ensaio imunoenzimático (EI) e reação em cadeia da polimerase (PCR). As espécies e genótipos de Cryptosporidium foram determinados pelo método de polimorfismo no tamanho dos fragmentos de restrição (RFLP) ou sequenciamento dos genes SSU-rRNA e GP60. A infecção por Cryptosporidium spp. teve ocorrência de 14% (28/200). A ocorrência em bezerros (26%) foi significativamente maior do que em vacas (2%). Do total de 27 amostras positivas submetidas à caracterização genética, C. andersoni foi identificado em 23 (85.1%), C. bovis em três (11.1%) e C. parvum subtipo IIaA15G2R1 em uma (3.7%). O presente estudo demonstrou que a infecção por Cyptosporidium é comum e difundida em bovinos leiteiros nessa região e que bezerros possuem uma alta prevalência de C. andersoni. A presença de C. parvum subtipo IIaA15G2R1 indica que bezerros leiteiros dessa região devem ser considerados uma fonte de oocistos de Cryptosporidium com potencial zoonótico.