Comparison of immunological and molecular methods for laboratory diagnosis of ocular toxoplasmosis in blood, serum and tears in Brazil.

Ocular toxoplasmosis (OT) is caused by protozoan T. gondii. Ophthalmological examination is considered the gold standard for OT diagnosis, and laboratory tests are used for diagnostic confirmation. However, these tests can present different results, which change depending on their basis, on sample type and on patients' clinical alteration. Thus, the aim of the present study is to assess immunodiagnostic and molecular techniques applied in blood, serum and tear fluid to diagnose T. gondii infection in patients seen at an Ophthalmology Clinic. In total, 160 patients were included in the study, 40 of them had OT with active lesions (G1); 40 had OT with healed lesions (G2), 40 had non-toxoplasmic uveitis (G3) and 40 had no ocular alterations (G4). Serum samples were subjected to Immunoenzymatic Assay (ELISA) and to Indirect Immunofluorescence Reaction (IFAT) to search for anti-T. gondii IgM and IgG. Tear fluid samples were analyzed through ELISA for IgA research. All blood and tear fluid samples were subjected to conventional polymerase chain reaction (cPCR) and in a Nested PCR model for T. gondii DNA amplification with targets B1, GRA7 and REP 529. IgG and IgM anti-T. gondii was detected in serum samples from 106 and 15 patients, respectively, when combining ELISA and IFAT results. Anti-T.gondii IgA antibodies were detected in 9.2% of the tear material. Nested PCR with GRA7 target showed higher positivity in blood samples (24.4%); Nested PCR with B1 target showed a higher frequency of positivity in tears (15%). Biological samples of patients with active lesions showed the highest positivity frequencies in all immunodiagnostic assays, as well as in most PCR models. The present results highlighted the need of associating techniques with different fundamentals to confirm OT diagnosis. Furthermore, further tear fluid analyses should be performed to validate this biological material as lesser invasive alternative for the more accurate OT diagnosis.

The Phylogenetic Characterization of Balantioides coli Isolated in the Pavlova Culture Medium Supplemented with Coconut Water and Animal Serum.

Balantioides coli is a ciliated protist that can cause dysentery in humans, pigs and nonhuman primates and may have the potential for zoonotic transmission. Its diagnosis is routinely performed through conventional parasitological techniques, and few studies have used culturing techniques to isolate it, applying molecular tools for the characterization of this protozoan. Thus, the objective of this study was to confirm B. coli diagnosis using molecular tools and to characterize the genetic variants of this parasite isolated from pigs kept on family farms in Brazil using three different culture media that differed in the serum added. Fecal samples from pigs were inoculated in Pavlova medium plus coconut water (PC), fetal bovine serum (PB) and horse serum (PH). Of the 127 samples positive for forms compatible with the phylum Ciliophora, 31 were selected for isolation. The most successful medium for isolation was PB 19/31 (61.3%), followed by PH 18/31 (58.1%) and PC 11/31 (35.5%). Of the nucleotide sequences generated, 20 were classified as genetic variant type B0, two as A1 and 15 as A0. The results indicated that PC, despite having allowed the isolation of B. coli for a short period, was not an adequate medium for the maintenance of this parasite in vitro, therefore requiring improvement.

Morphological and molecular characterization of parabasilids isolated from ex situ nonhuman primates and their keepers at different institutions in Brazil.

Intestinal protozoa, which can be asymptomatic or cause diarrhea, dysentery and even death, are among the main agents that affect nonhuman primates (NHPs) kept under human care. Nevertheless, information on the molecular and morphometric profiles of parabasilids in the Neotropics is still scarce. In this context, the objective of this study was to isolate the Parabasalia protozoa detected in the feces of NHPs and their keepers in Pavlova and TYSGM9 media and to characterize the isolates by molecular biology and morphometry. Fecal samples from NHPs from five Brazilian institutions were analyzed. Direct examination was performed immediately after obtaining the samples. A total of 511 fecal samples from NHPs were collected, and 10.6% contained parabasilids. Regarding the handlers, of the 74 samples analyzed, three were positive. In vitro-generated parabasilid isolates were successfully obtained from all positive samples, as identified via microscopy. Isolates of the parasite were obtained both from New World NHPs, including the genera Leontopithecus, Saguinus, Leontocebus, Aotus, Saimiri, Sapajus, and Alouatta, and from the Old World primate Pan troglodytes. Forty-nine NHP isolates were molecularly identified: Pentatrichomonas hominis (16), Trichomitus batrachorum (14), Tetratrichomonas brumpti (13) and Hypotrichomonas hampli (6). The human isolates were identified as Tetratrichomonas sp. (2) and T. batrachorum (1). Visualization and morphometric analysis revealed trophozoites with piriform or rounded shapes that presented variable measurements. The isolates previously characterized as P. hominis had up to five free flagella, while T. batrachorum and Tetratrichomonas sp. had up to four free flagella, and H. hampli had a maximum of three free flagella. These morphometric characteristics corroborated the molecular identification. In general, a variety of parabasilids were observed to infect NHPs, and T. batrachorum was isolated from biological samples from both NHPs and their keepers, a finding that reinforces the susceptibility of these hosts to infections by parabasilids in Brazil.

Occurrence of Toxoplasma gondii and other gastrointestinal parasites in free-roaming cats from the Rio de Janeiro zoo.

This communication aimed to determine the frequency of infection by T. gondii and other gastrointestinal parasites in free-living cats captured in BioParque do Rio (Fundação RioZoo), Brazil. To this end, 58 blood and 51 fecal samples were collected from 68 cats from July 2019 to September 2020. The serum obtained was analyzed by indirect fluorescent antibody test for anti-T. gondii IgG. Fecal samples were examined by microscopic parasitological techniques. Of the total, 10.3% of the animals presented anti-T. gondii IgG. Parasitic structures were observed in 39.2% of the fecal samples. Hookworms were the most detected parasites (19.6%) followed by Cystoisospora sp. (11.7%), Dipylidium caninum (7.8%) and Toxocara cati (3.9%). These results indicated the exposure of this population of stray cats to potentially zoonotic parasites, which in addition to causing possible damage to domestic and wild animal health, also pose risks to public health.

Esta comunicação teve como objetivo determinar a frequência de infecção por T. gondii e outros parasitas gastrointestinais em gatos de vida livre capturados no BioParque do Rio (Fundação RioZoo), Brasil. Para isso, foram coletadas 58 amostras de sangue e 51 amostras fecais de 68 gatos capturados entre julho de 2019 e setembro de 2020. O soro obtido foi submetido à reação de imunofluorescência indireta para IgG anti-T. gondii. As amostras fecais foram submetidas a técnicas parasitológicas microscópicas. Do total, 10,3% dos animais apresentaram IgG anti-T. gondii. Estruturas parasitárias foram observadas em 39,2% das amostras fecais. Os ancilostomídeos foram os parasitas mais detectados (19,6%), seguidos por Cystoisospora sp. (11,7%), Dipylidium caninum (7,8%) e Toxocara cati (3,9%). Esses resultados indicaram a exposição dessa população de gatos de rua a parasitas potencialmente zoonóticos, que, além de causar possíveis danos à saúde dos animais domésticos e selvagens, também representam riscos à saúde pública.

Toxoplasma gondii in the faeces of wild felids from the Atlantic Forest, Brazil

BACKGROUND Toxoplasma gondii is a apicomplexan parasite of virtually all warm-blooded species. All true cats (Felidae) can act as definitive hosts for this parasite by shedding resistant oocysts into the environment. However, the patterns of oocysts shedding are only partially understood in domestic cats and largely unknown in wild felids. OBJECTIVES We carried out molecular analysis of 82 faecal samples from wild felids collected in the Serra dos Órgãos National Park (Parnaso), Rio de Janeiro, Brazil. METHODS We screened samples for T. gondii DNA using a quantitative polymerase chain reaction (qPCR) targeting the 529bp DNA fragment. Polymerase chain reaction (PCR)-positive samples were genotyped using 15 microsatellite markers. RESULTS Only one faecal sample from a Puma yagouaroundi was PCR-positive [cycle threshold (Ct) = 26.88]. This sample was contaminated by a T. gondii strain of BrIII lineage, a common lineage in domestic animals from Brazil. MAIN CONCLUSIONS This first report of T. gondii in faeces of wild South American felids in their natural environment indicates infrequent oocyst shedding and suggests a role of acquired immunity in limiting re-excretion as in domestic cats. The presence of a domestic strain of T. gondii in a faecal sample from a wild felid at very low concentrations (not detected by microscopy) is consistent with the hypothesis of host-parasite co-adaptations limiting the circulation of T. gondii strains between domestic and wild environments.

Toxoplasma gondii among captive wild mammals in zoos in Brazil and Cuba: seroprevalence and associated risk factors / Toxoplasma gondii entre mamíferos silvestres cativos em zoológicos no Brasil e em Cuba: soroprevalência e fatores de risco associados

Abstract Toxoplasma gondii is a zoonotic parasite of worldwide distribution that can infect several species of homeothermic animals. Few studies have evaluated the exposure of captive wild animals to T. gondii. This study involved a serological survey of anti-T. gondii antibodies in mammals kept in Cuba's National Zoo (PZN) and in the Rio de Janeiro Zoo (RIOZOO) in Brazil. The study consisted of a total of 231 serum samples from mammals, 108 from PZN and 123 from RIOZOO. All the samples were subjected to IgG anti-T. gondii testing by means of the inhibition ELISA method and the modified agglutination test, respectively. T. gondii antibodies were detected in 85.2% samples from PZN and 32.5% samples from RIOZOO. At the PZN, Perissodactyla (92.3%) was the order with the highest serological prevalence rate, whereas at the RIOZOO, the order Primates (46.7%) stood out (p<0.05). In addition to this association, the origin of the PZN animals was also associated with T. gondii infection. This finding demonstrates the need for constant veterinary monitoring of captive wild mammals in order to link the serological diagnosis with clinical alterations indicative of toxoplasmosis.

Resumo Toxoplasma gondii é um parasito zoonótico de distribuição mundial que pode infectar várias espécies de animais homeotérmicos. Poucos estudos avaliaram a exposição de animais silvestres em cativeiro ao T. gondii. Este estudo envolveu uma pesquisa sorológica de anticorpos anti-T. gondii em mamíferos mantidos no Zoológico Nacional de Cuba (PZN) e no Zoológico do Rio de Janeiro (RIOZOO) no Brasil. O estudo consistiu em um total de 231 amostras de soro de mamíferos, sendo 108 do PZN e 123 do RIOZOO. Todas as amostras foram submetidas à pesquisa de IgG anti-T. gondii pelos métodos de ELISA de inibição (PZN) e teste de aglutinação modificado (RIOZOO). Anticorpos de T. gondii foram detectados em 85,2% das amostras do PZN e 32,5% das amostras do RIOZOO. No PZN, Perissodactyla (92,3%) foi a ordem com maior taxa de prevalência sorológica, enquanto no RIOZOO a ordem Primatas (46,7%) se destacou (p <0,05). Além dessa associação, a origem dos animais PZN também foi associada à infecção por T. gondii. Esse achado demonstra a necessidade de monitoramento veterinário constante de mamíferos silvestres em cativeiro, a fim de vincular o diagnóstico sorológico a alterações clínicas indicativas de toxoplasmose.

Toxoplasma gondii and other parasites studied in feline feces in Zoos in Cuba and Brazil

Under certain circumstances, wild animals kept in zoos may be more exposed to infectious parasitic diseases. The puprpose of this study was to determine the frequency of gastrointestinal parasites in captive wild felids in the National Zoological Park (PZN) in Cuba (Havana) and in RioZoo in Brazil (Rio de Janeiro). A total of 52 fecal samples were collected from 52 felids, as follows: 19 Panthera leo, two Leopardus tigrinus, two Leopardus pardalis, one Panthera tigris altaica, four Panthera tigris tigris, six Panthera onca, seven Puma concolor, one Herpailurus yagouaroundi, three Acinonyx jubatus, two Caracal caracal and five Panthera pardus. The fecal samples were processed and examined microscopically. The frequency of parasite positive animals was 17.5% (7/40) in PZN and 25% (3/12) in RioZoo. Panthera pardus (40%) and Panthera onca (20%) were most frequently infected in PZN and Panthera leo (100%) and Leopardus pardalis (50%) in RioZoo. Hookworm (12.5%) was detected in PZN as well as Toxascaris leonina (10%) and in RioZoo nematode larvae (9.1%) , hookworm eggs (9.1%), Toxascaris leonina (2%) and the cestode eggs from the Diphyllobothriidae family (9.1%) were found. Toxoplasma gondii-like oocysts were not detected in feline feces. Although the positivity of gastrointestinal parasites detected in feline fecal samples was not very high in these zoos, both institutions need to implement and maintain sanitary measures, including routine diagnosis of parasitosis followed by specific treatment according to the infections detected.

Toxoplasma gondii: infection among shelter and stray cats in Rio de Janeiro, Brazil / Toxoplasma gondii: infecção entre os gatos de abrigo e de rua no Rio de Janeiro, Brasil

Abstract The aim of this study was to identify possible infection of Toxoplasma gondii among cats in a shelter and a set of condominiums in the city of Rio de Janeiro, through changes to the cats' serological status between two different times in 2014 and 2015. One group was made up of captive cats at the municipal shelter and the other comprised stray cats that circulated in condominiums in the city. On the first occasion, cats were caught and tagged through application of microchips; in this manner, blood samples were obtained from 261 captive cats and 172 stray cats. On the second occasion, blood samples were obtained from 94 captive cats and 56 recaptured stray cats. The serological diagnosis was made by means of the indirect hemagglutination assay (IHA) and indirect immunofluorescence reaction (IFAT) (cutoff ≥ 64). The frequency of T. gondii infection among the captive cats was 24.5% and among the stray cats, 18%. With the second analysis, it was possible to verify modifications to the serological status of anti-T. gondii antibodies, in 18% of both populations of animals. The presence of seroconversion shows that infection was possibly occurring in the region at the time of the study.

Resumo O objetivo deste estudo foi identificar uma possível infecção por Toxoplasma gondii entre gatos de abrigo e de um conjunto de condomínios na cidade do Rio de Janeiro, por meio de mudanças no status sorológico dos gatos em dois momentos diferentes em 2014 e 2015. O grupo foi formado por gatos, denominados cativos, de um abrigo municipal, e o outro por gatos de rua que circulavam em condomínios da cidade. Na primeira ocasião, os gatos foram capturados, microchipados e coletadas amostras de sangue de 261 gatos cativos e de 172 gatos de rua. Na segunda ocasião, as amostras de sangue foram obtidas de 94 gatos cativos e 56 de gatos de rua recapturados. O diagnóstico sorológico foi realizado por meio do ensaio de hemaglutinação indireta (HAI) e pela reação de imunofluorescência indireta (RIFI) (ponto de corte ≥ 64). A frequência de infecção por T. gondii entre os gatos cativos foi de 24,5% e entre os gatos de rua 18%. Com a segunda análise, foi possível verificar modificações no status sorológico de anticorpos anti-T. gondii, em 18% em ambas populações de animais. A presença de soroconversão mostra que a infecção possivelmente ocorreu na região no momento do estudo.

Comparative study of three xenic media culture for cultivation of Balantidium coli strains / Estudo comparativo entre três meios de cultivo xênicos para o cultivo de cepas de Balantidium coli

Abstract The aim of the present study was to evaluate the growth rate of Balantidium coli in three xenic media cultures. Between 2013 and 2015, 10 B. coli isolates obtained from feces of Cynomolgus macaques, and 30 isolates from feces of pigs were studied. An inoculum of 500 trophozoites was transferred to tubes containing LES, TYSGM-9 and Pavlova media. These cultures were evaluated at incubation times of 24, 48, 72 and 96 hours. In most of strains analyzed wasn't showed significant difference in the growth rate comparing TYSGM-9 and Pavlova media (Wilcoxon p>0.016). In Pavlova medium, the trophozoites showed a maximum growth at 72 hours with significant difference when compared with the times of 24 h and 96 h (Wilcoxon <0.008). In LES, viable trophozoites were observed until 24 hours, with a significant difference (Friedman p<0.05, Wilcoxon p<0.016) in the number of parasite cells compared with Pavlova and TYSGM-9 media cultures. Thus, LES medium seemed to be less adequate than the other media for maintenance of B. coli. Despite the satisfactory results in TYSGM-9, Pavlova medium was considered ideal for the maintenance of this protozoan strain, guaranteeing the viability of the parasite with subculture every three days, presenting lower costs.

Resumo O objetivo do presente estudo foi avaliar a taxa de crescimento de Balantidium coli em três meios de cultura xênicos. Entre 2013 e 2015 foram estudados 10 isolados de B. coli obtidos de Cynomolgus macaques e 30 isolados de suínos. Um inóculo contendo 500 trofozoítos foi transferido para tubos contendo os meios LES, TYSGM-9 e Pavlova. Os cultivos foram avaliados com tempos de incubação de 24, 48, 72 e 96 horas. Na maioria das cepas analisadas não foi observado diferença significativa na taxa de crescimento comparando TYSGM-9 e Pavlova (Wilcoxon p>0,016). Em Pavlova, os trofozoítos apresentaram máximo de crescimento a 72 h com diferença significativa quando se comparou com os tempos de 24 h e 96 h (Wilcoxon <0,008). Em LES observou-se trofozoítos viáveis até 24 horas com diferença significativa (Friedman p<0,05 e Wilcoxon p<0,016), na quantidade de células parasitárias, quando comparado com Pavlova e TYSGM-9. Dessa forma, o meio LES mostrou-se ser menos adequado do que os outros, para a manutenção de B. coli. Apesar do resultado satisfatório em TYSGM-9, Pavlova foi considerado ideal para manutenção do protozoário, por garantir a viabilidade do parasito com subcultivos a cada três dias, além de apresentar menor custo.

Gastrointestinal parasites among felids inhabiting the Serra dos Órgãos National Park, Rio de Janeiro, Brazil / Parasitos gastrintestinais em felídeos que habitam o Parque Nacional da Serra dos Órgãos, Rio de Janeiro, Brasil

Abstract This study aimed to investigate the species of felids that inhabit the Serra dos Órgãos National Park (Parnaso) and gastrointestinal parasites at various stages of their life cycles in the feces of these animals. Between 2013 and 2015, felid feces were collected from trails in Parnaso. The sampling points were georeferenced. A total of 82 fecal samples were processed, of which 79 were collected on the ground, two from captured felids and one from a necropsied animal. All samples underwent coproparasitological techniques. Samples collected from the environment underwent additional trichological analysis. Fur patterns corresponding to Leopardus guttulus, Leopardus pardalis, Leopardus wiedii and Puma yagouaroundi were observed in 32 of the samples collected on the soil. High frequency of potentially parasitic evolving forms (88.6%) was observed in felid feces, particularly eggs of the family Diphyllobothriidae (68.6%). Besides, were also detected, eggs of superfamily Ascaridoidea (42.9%), nematode larvae (28.6%), eggs of order Strongylida (28.6%), Capillaria sp. (8.6%), Trichuris sp. (8.6%), eggs of order Spirurida (2.9%), unsporulated coccidian oocysts (8.6%) and Eimeria sp. (2.9%). Felid feces presented higher frequency of polyparasitism (60%) than monoparasitism (28.6%).

Resumo Este estudo teve como objetivo pesquisar as espécies de felídeos que circulam no Parque Nacional da Serra dos Órgãos (Parnaso), bem como as formas evolutivas de parasitos gastrintestinais nas fezes desses animais. Entre 2013 e 2015, fezes de felídeos foram coletadas nas trilhas do Parnaso, sendo os pontos de coleta georeferenciados. Foram coletadas 82 amostras fecais, sendo 79 do ambiente, duas de felídeos capturados e uma de um animal necropsiado. Todas as amostras foram submetidas a técnicas coproparasitológicas, sendo as coletadas do ambiente também analisadas pela tricologia. Em 32 amostras coletadas do solo foram identificados padrões de pelos de Leopardus guttulus , Leopardus pardalis, Leopardus wiedii e Puma yagouaroundi. Elevada frequência de formas evolutivas potencialmente parasitárias (88,6%) foi evidenciada nas fezes dos felídeos, destacando os ovos da Família Diphyllobotrhiidae (68,6%). Além destes, também foram detectados, ovos da superfamília Ascaridoidea (42,9%), larvas de nematoides (28,6%), ovos da ordem Strongylida (28,6%), Capillaria sp. (8,6%), Trichuris sp. (8,6%), ovos da família Spirurida (2,9%), oocistos de coccídios não esporulados (8,6%) e Eimeria sp. (2,9%). As fezes dos felídeos apresentaram maior frequência de poliparasitismo (60%) que de monoparasitismo (28,6%).

Gastrointestinal parasites in stray and shelter cats in the municipality of Rio de Janeiro, Brazil / Parasitos gastrintestinais de gatos de rua e de abrigo da cidade do Rio de Janeiro, Brasil

Abstract The increasingly urban nature of the population has led many people to choose independent pets, such as cats. This situation has also made it possible for these animals to be abandoned, thus increasing the numbers of cats on the streets and in shelters. These animals can act as a source of infection for other hosts. Between 2014 and 2015, the frequency of gastrointestinal parasites in captive and stray cats in the municipality of Rio de Janeiro was analyzed. Ninety-one fecal samples were collected from captive cats and 172 from stray cats. Centrifugal sedimentation and flotation techniques were used. The frequency of parasites among the stray cats was 77.3%, and this was significantly higher than the frequency observed in captive cats (49.5%). Helminths were detected more frequently, and hookworms were the parasites most detected. Toxocara cati, Cystoisospora sp. and Dipylidium caninum were also detected. No statistical difference in the frequency of parasites was observed between the sexes among the captive cats. However, among the stray cats, males (85.5%) presented higher positivity than females (71.8%). The high frequency of hookworms, which are the agent for "cutaneous larva migrans" in humans, shows the need to control parasitic infections among the cats studied.

Resumo O aumento da população urbana contribuiu para que muitas pessoas escolhessem um animal de estimação independente, como o gato. Esta situação possibilita o abandono desses animais, aumentando o número de gatos nas ruas e nos abrigos. Esses animais, então, podem servir como fonte de infecção para outros hospedeiros. Entre 2014 a 2015 foi analisada a frequência de parasitos gastrintestinais em gatos cativos e de rua na cidade do Rio de Janeiro. Foram coletadas 91 amostras fecais de gatos cativos e 172 de gatos de rua, sendo essas processadas pelas técnicas de centrífugo sedimentação e flutuação. A frequência de parasitos nos gatos de rua foi 77,3%, sendo esta significativamente maior do que a evidenciada nos cativos, 49,5%. Os helmintos foram mais detectados, sendo ancilostomídeos os parasitos mais evidenciados. Também foram detectados, Toxocara cati, Cystoisospora sp. e Dipylidium caninum. Não foi observada diferença estatística na frequência de parasitos entre o sexo nos gatos cativos, porém nos de rua, os machos (85,5%) apresentaram positividade maior do que as fêmeas (71,8%). A elevada frequência de ancilostomídeos, agente da "Larva Migrans Cutânea" no homem, demonstra a necessidade de controle das infecções parasitárias nos gatos estudados.

Intestinal parasites in children with neurological disorders treated at a rehabilitation institution in Niterói, Rio de Janeiro, Brazil

Enteroparasitosis presents high prevalence in developing countries, and is frequent among children, and those with disabilities present even higher susceptibility. The present study evaluated 156 children with neurological disorders to whose parents or guardians an epidemiological questionnaire was applied with a view to identifying risk factors. Stool samples were processed using Ritchie modified by Young, Faust et al., Lutz and Baermann-Moraes techniques. Thirteen samples (8.3%) were positive, being Blastocystis sp. the most frequent parasite (46.2%). Higher positivity was observed for protozoa, especially non-pathogenic. A statistically significant association was only found between the educational level of the mother and positivity for enteroparasites. Among positive children, the habit of placing their hands and objects in their mouths and the consumption of raw vegetables was more frequent. The results show the existence of care and sanitation minimizing the acquisition of intestinal parasite diseases. It is important to alert parents and caregivers about personal hygiene, water and food quality, as well as the environment, considering that transmission of these parasites occurs via the fecal-oral route, in association with multiple factors.

Avaliação da frequência de Balantidium coli em suínos, tratadores de suínos e primatas não humanos no estado do Rio de Janeiro / Balantidium coli: frequency evaluation in pig, pig handlers and nonhuman primate in the state of Rio de Janeiro, Brazil

Este trabalho relata a frequência de Balantidium coli nos tratadores de suínos e primatas não humanos que trabalham em criatórios localizados em distintos municípios do estado do Rio de Janeiro. No período de 2012 a 2014, foram coletadas 104 amostras fecais de 52 tratadores, 790 amostras fecais de suínos e 1.115 de primatas não humanos. Todas as amostras foram coletadas sem conservante químico, sendo submetidas às técnicas de Faust et al., Sheather modificada, Ritchie modificada, Lutz e exame direto. Formas evolutivas de B. coli não foram detectadas nas amostras fecais dos tratadores, porém foram evidenciadas em 49,2% dos suínos e em 41,5% do material fecal dos símios. A ausência da balantidiose em humanos, embora estivessem em contato próximo com animais parasitados, como foi demonstrado neste estudo, pode ter ocorrido em virtude da adoção de adequadas medidas higiênico-sanitárias na manipulação desses animais ou graças à resistência humana à infecção por esse protozoário.

Comparison of five parasitological techniques for laboratory diagnosis of Balantidium coli cysts / Comparação de cinco técnicas parasitológicas para diagnóstico laboratorial de cistos de Balantidium coli

Abstract Balantidium coli is a protozoon that can cause dysentery in humans, pigs and nonhuman primates, with zoonotic potential. In the literature, there is still little information on the effectiveness of different laboratory techniques for diagnosing this disease. This study compared and evaluated the performance of the Lutz, modified Ritchie, Faust, modified Sheather and direct examination techniques for detecting cysts of this protozoon. Between 2012 and 2014, 1905 fecal samples were collected from captive animals in the state of Rio de Janeiro. Of these, 790 were obtained from the rectum of pigs and 1115 from enclosures occupied by nonhuman primates. B. coli cysts were most evident through direct examination (22.4% of the samples) and the Lutz technique (21%). Fair agreement (Kappa = 0.41; p < 0.05) was observed only between direct examination and Lutz. The flotation techniques (Faust and modified Sheather) did not show good recovery of cysts. A statistically significant difference (p < 0.05) in the frequency of cysts between pigs and nonhuman primates could only be observed through direct examination and the Lutz technique. The most efficient method for diagnosing this parasitosis was seen to an association between direct examination and the spontaneous sedimentation technique.

Resumo Balantidium coli é um protozoário que pode determinar disenteria em humanos, suínos e primatas não humanos apresentando potencial zoonótico. Na literatura ainda são escassas as informações sobre a eficiência das diferentes técnicas laboratoriais para o diagnóstico dessa parasitose. Este estudo comparou e avaliou o desempenho das técnicas de Lutz, Ritchie modificada, Faust, Sheather modificada e do exame direto para a detecção de cistos desse protozoário. Entre 2012 e 2014, foram coletadas 1905 amostras fecais de animais cativos no Estado do Rio de Janeiro. Dessas, 790 foram obtidas da ampola retal de suínos e 1115 dos recintos de primatas não humanos. Cistos de B. coli foram 22,4 % mais evidenciados pelo exame direto; e pela técnica de Lutz, 21% das amostras. Concordância regular (Kappa = 0,41; p < 0,05) foi observada somente entre exame direto e Lutz. As técnicas de flutuação, Faust et al. e Sheather modificada não apresentaram boa recuperação dos cistos. Diferença estatística significativa (p < 0,05) na frequência de cistos entre suínos e primatas não humanos pode ser observada somente no exame direto e na técnica de Lutz. A metodologia mais eficiente para diagnóstico dessa parasitose foi observada pela associação do exame direto e da técnica de sedimentação espontânea.

Conhecimentos de escolares e funcionários da Rede Pública de Ensino sobre as parasitoses intestinais / Knowledge of students and employees of Public School System on the intestinal parasites

O desconhecimento ou nível insatisfatório de informações sobre as enteroparasitoses e sua prevenção, aliados ao baixo nível socioeconômico, educacional e insuficiência de saneamento básico, têm sido apontados como fatores de risco para a aquisição dessas infecções. Neste estudo foram averiguados os conhecimentos de estudantes do 3º ao 7º ano do Ensino Fundamental e dos funcionários de sete escolas Municipais de Niterói, RJ, Brasil. Foram aplicados questionários, e as respostas foram avaliadas como adequadas, inadequadas, incompletas, não soube responder ou não respondeu. Entre os 335 estudantes, observou-se conhecimento mediano sobre habitat (47,5 %), prevenção (48,4 %) e sintomatologia (45,1 %). Desses, 43,3 % demonstraram conhecimento inadequado sobre transmissão. E 51,6 % dos estudantes fizeram associação de verminoses com vermes, e somente 2,7 % apresentaram conhecimento sobre protozoários. Em 62 funcionários, observou-se conhecimento adequado sobre definição, exemplos, habitat, transmissão, sintomatologia e prevenção. Tais resultados evidenciaram a ocorrência de informações fragmentadas sobre a infecção por enteroparasitos entre os escolares e mais corretas entre os funcionários. Torna-se importante efetuar melhor abordagem do tema, referente aos conteúdos de ciências, e ampliar este arcabouço teórico para interferir na difusão da informação e na formação de cidadãos críticos.

Lack of knowledge or unsatisfactory information about the enteroparasites and preventive measures, combined with low socioeconomic, educational and poor sanitation status, have been appointed as a risk factor for acquiring these parasites. This study aimed at identifying the knowledge of students of 3rd to 7th grade of elementary school and of employees of seven municipal schools of Niterói, RJ, Brazil. Questionnaires were applied, and the answers were rated as adequate, inadequate, incomplete, could not answer or did not answer. Among 335 students, the median knowledge on habitat (47.5 %), prevention (48.4 %) and symptoms (45.1 %) were detected. Of these, 43.3 % showed an inadequate knowledge on transmission; 51.6 % associated the verminosis with worms, and 2.7 % only had knowledge about protozoa. The adequate knowledge on definition, examples, habitat, transmission, symptoms and prevention were observed among 62 employees. Occurrence of fragmented information on the enteroparasites among students and the correct knowledge among employees. Thus, it is crucial to perform the better approach, referring to science content, aiming to promote the expansion of this theoretical framework for interfering with the information dissemination and in the development of critical citizens.

Conhecimentos de escolares e funcionários da rede pública de Ensino sobre as parasitoses intestinais / Knowledge of students and employees of public school system on the intestinal parasites

O desconhecimento ou nível insatisfatório de informações sobre as enteroparasitoses e sua prevenção, aliados ao baixo nível socioeconômico, educacional e insuficiência de saneamento básico, têm sido apontados como fatores de risco para a aquisição dessas infecções. Neste estudo foram averiguados os conhecimentos de estudantes do 3º ao 7° ano do Ensino Fundamental e dos funcionários de sete escolas Municipais de Niterói, RJ, Brasil. Foram aplicados questionários, e as respostas foram avaliadas como adequadas, inadequadas, incompletas, não soube responder ou não respondeu. Entre os 335 estudantes, observou-se conhecimento mediano sobre habitat (47,5 %), prevenção (48,4 %) e sintomatologia (45,1 %). Desses, 43,3 % demonstraram conhecimento inadequado sobre transmissão. E 51,6 % dos estudantes fizeram associação de verminoses com vermes, e somente 2,7 % apresentaram conhecimento sobre protozoários. Em 62 funcionários, observou-se conhecimento adequado sobre definição, exemplos, habitat, transmissão, sintomatologia e prevenção. Tais resultados evidenciaram a ocorrência de informações fragmentadas sobre a infecção por enteroparasitos entre os escolares e mais corretas entre os funcionários. Torna-se importante efetuar melhor abordagem do tema, referente aos conteúdos de ciências, e ampliar este arcabouço teórico para interferir na difusão da informação e na formação de cidadãos críticos.

Lack of knowledge or unsatisfactory information about the enteroparasites and preventivemeasures, combined with low socioeconomic, educational and poor sanitation status, havebeen appointed as a risk factor for acquiring these parasites. This study aimed at identifying the knowledge of students of 3rd to 7th grade of elementary school and of employees of seven municipal schools of Niterói, RJ, Brazil. Questionnaires were applied, and the answers were ratedas adequate, inadequate, incomplete, could not answer or did not answer. Among 335 students,the median knowledge on habitat (47.5 %), prevention (48.4 %) and symptoms (45.1 %) were detected. Of these, 43.3 % showed an inadequate knowledge on transmission; 51.6 % associatedthe verminosis with worms, and 2.7 % only had knowledge about protozoa. The adequate knowledge on definition, examples, habitat, transmission, symptoms and prevention were observed among62 employees. Occurrence of fragmented information on the enteroparasites among studentsand the correct knowledge among employees. Thus, it is crucial to perform the better approach, referring to science content, aiming to promote the expansion of this theoretical frameworkfor interfering with the information dissemination and in the development of critical citizens.

Parasitos intestinais em alfaces (lactuca sativa, L) das variedades crespa e lisa comercializadas em feiras livres em Niterói-RJ / Intestinal parasites in curly and smooth lettuce (lactuca sativa, L. ) commercialized in free markets from Niterói-RJ

Foram analisadas 210 amostras de alface comercializadas em oito feiras livres de Niterói-RJ, sendo 105 da variedade crespa e 105 da lisa. Foi detectada contaminação por parasitos em 12/210 (5,7 porcento) das amostras, sendo 4,8 porcento em amostras crespas e 6,7 porcento em lisas. Em 42,9 porcento das amostras crespas e 38,1 porcento das lisas, foram encontrados ovos de nematoides da superfamília Rhabditoidea e larvas de nematoides em vida livre. Artrópodes e adultos de nematoides estavam presentes em 41,9 porcento (88/210) das amostras, sendo 53,3 porcento em amostras crespas e 30,5 porcento em lisas. Não houve diferença estatisticamente significativa entre as variedades de alface quanto à presença de contaminantes. A existência de contaminantes de origem biológica indica qualidade higiênica insatisfatória das alfaces comercializadas nas feiras livres de Niterói-RJ e o consumo de ambas as variedades da hortaliça, sem lavagem adequada, caracteriza risco de aquisição de enteroparasitos...

Two hundred and ten samples of lettuce, sold in eight free markets from Niterói-RJ, were analyzed for intestinal parasite contamination. Contamination by evolutive forms of parasites was detected in 12/210 (5.7 percent) of the samples, being 4.8 percent in thecurly variety and 6.7 percent in smooth samples. Nematode eggs of the superfamily Rhabditoidea and free living larvae of nematodes were found in 42.9 percent of curly samples and 38.1 percent of smooth ones. Adult arthropods and nematodes were found in 41.9 percent (88/210) of samples, 53.3 percent of curly samples and 30.5 percent of smooth samples. There was no statistically significant difference between the varieties of lettuce relative to the presence of contaminants. The presence of contaminants of biological origin indicates poor hygienic quality of lettuce sold in free markets of Niterói-RJ, representing a risk for infection by intestinal parasites, if consumed without proper washing, in both varieties...

Parasitological and immunological diagnoses from feces of captive-bred snakes at Vital Brazil Institute / Diagnóstico parasitológico e imunológico em fezes de serpentes cativas do Instituto Vital Brazil

Fecal samples from 56 snakes at the Vital Brazil Institute, in the city of Niterói, Rio de Janeiro, were tested using the sedimentation and flotation techniques to investigate the evolutionary forms of parasites such as helminths and protozoa, and using enzyme immunoassay techniques to detect antigens of Cryptosporidium sp. and Giardia sp. Among the animals tested, 80.3% were positive for parasites. Out of these, there were 16 Bothrops jararaca, 16 B. jararacussu and 13 Crotalus durissus. The prevalence of parasitic nematodes was 41.1%, and nematodes were found in all three snake species. Among these, the most frequent finding was eggs of Kalicephalus sp., which were diagnosed in 25% of the snakes. The positivity for protozoa detected using parasite concentration techniques was 75%, including oocysts of Caryospora sp. in 75%, cysts with morphology similar to Giardia sp. 3.6%, amoeboid cysts in 41.1% and unsporulated coccidia oocysts in 8.9%. Immunoassays for Cryptosporidium sp. antigens produced positive findings in 60.7%. Pseudoparasites were detected in 64.3%. These results show that there is a need to improve the sanitary handling of captive-bred snakes, and also for the animal house that supplies rodents to feed them. The results also highlight that diagnostic tests should be performed periodically on stool specimens from captive-bred snakes.

Foram analisadas amostras fecais de 56 serpentes do Instituto Vital Brazil, Niterói, Rio de Janeiro, por técnicas de sedimentação e flutuação, para pesquisa de formas evolutivas de parasitos como helmintos e protozoários, e imunoenzimáticas para detecção de antígenos de Cryptosporidium sp. e Giardia sp. Dos animais estudados, 80,3% foram positivos para parasitos, dos quais 16 pertenciam à espécie Bothrops jararaca, 16 à B. jararacussu e 13 à Crotalus durissus. A prevalência de nematódeos parasitos foi de 41,1%, sendo evidenciados nas três espécies de serpentes. Dentre estes, o mais frequente foi o encontro de ovos de Kalicephalus sp., diagnosticado em 25% das serpentes. A positividade para protozoários detectados pelas técnicas de concentração foi de 75%, nas quais oocistos de Caryospora sp. foram detectados em 75%, cistos com morfologia similar a Giardia sp. em 3,6%, cistos de amebídeos em 41,1% e oocistos não esporulados de coccídios em 8,9%. Obteve-se positividade para antígenos de Cryptosporidium sp. por imunoensaio em 60,7%. Pseudoparasitos foram detectados em 64,3%. Estes resultados apontam para a necessidade de aprimoramento do manejo sanitário das serpentes cativas, bem como no biotério que fornece roedores para sua alimentação, destacando a realização periódica de diagnóstico nas amostras fecais de serpentes cativas.

Avaliação parasitológica da água de abastecimento e do soloperidomiciliar de Aldeias Guarani / Parasitological assessment of the water supply and the domiciliary surrounding area soil in Guarani Villages

As aldeias Guarani de Angra dos Reis e Paraty possuem abastecimento de água constituído por captaçãodas nascentes, tratamento da água com cloro e/ou filtros e reservatórios de armazenamento. O sistema desaneamento é estruturado por módulos sanitários fornecidos pela Funasa. Para investigar a contaminaçãoambiental por protozoários e helmintos, as amostras de água e de solo das aldeias foram avaliadas pormeio de análises parasitológicas. De quatro aldeias, foram coletadas 24 amostras de água nas captaçõescom cartuchos filtrantes, 24 dos reservatórios de água clorada por sifonação e 64 amostras de solo porraspagem superficial no peridomicílio. Todas as amostras foram analisadas pelas técnicas de centrífugosedimentação,centrífugo-flutuação e ensaio imunoenzimático para Cryptosporidium spp., Giardia lambliae Entamoeba histolytica, associando-se à pesquisa de larvas de nematódeos no solo. As aldeias de ParatyMirim e Rio Pequeno apresentaram maior positividade para parasitas nas amostras de água, detectandosecistos de amebídeos e Giardia sp., antígenos de G. lamblia, E. histolytica e Cryptosporidium spp. Foramdetectados ovos de Ascaris sp., Trichuris sp., antígenos de Cryptosporidium spp. e E. histolytica nas amostrasde solo da aldeia Sapukai. Esses resultados indicam elevada contaminação ambiental e deficiência notratamento da água que abastece as aldeias Guarani.