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.

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.

Prevalence, Risk Factors and Diagnosis of Helminths in Thoroughbred Horses Kept at Training Centers in Rio de Janeiro, Brazil.

The aims of this study were to determine the prevalence of helminths in Thoroughbred horses in Rio de Janeiro; make correlations with risk factors for these infections; and compare the efficiency of three floatation solutions applied in the quantitative Mini-FLOTAC technique. Fecal samples from 520 horses were collected from six training centers between 2019 and 2021. These were subjected to the Mini-FLOTAC technique using three solutions: NaCl (density = 1.200 g/mL), ZnSO4 (1.350 g/mL) and ZnSO4 (1.200 g/mL); and also to qualitative techniques. Information on the horses' sex and age of horses was retrieved from the studbook; data on management from a questionnaire. The overall prevalence of intestinal parasites was 71.9%, with significant differences between training centers (P ≤ .05). On farm C, 87.7% of the samples presented strongylids and 38.7% had Parascaris spp., with the highest egg counts per gram of feces (EPG), of 358.33 and 40.41 respectively. Horses less than 3 years of age were about eight times more likely to be parasitized by strongylids and eleven times more likely to have EPG ≥500. The NaCl solution used in Mini-FLOTAC enabled recovery of the greatest number of samples with high EPG and reached the highest sensitivity values in the diagnosis when compared to the other solutions. Moreover, in the diagnoses, the levels of agreement between the results from the solutions used in Mini-FLOTAC were substantial. However, in estimating the EPG, full agreement between the results from the solutions used in Mini-FLOTAC was not obtained.

First microscopic and molecular parasitological survey of Strongylus vulgaris in Brazilian ponies.

The frequency of gastrointestinal parasites with an emphasis on Strongylus vulgaris was investigated among the Brazilian Pony breed kept on farms in the municipality of Teresópolis, state of Rio de Janeiro. Fecal samples were collected in three stud farms: A (n= 22 animals), B (n= 3), and C (n= 2). Fecal samples were subjected to the quantitative Mini-FLOTAC technique, using three different solutions, and to qualitative techniques. The parasite prevalence was found to be 81.4%. Eggs from strongylids were identified in 74% of the ponies. Eggs of Parascaris spp. were detected in 22.7% of the animals, which were all females of farm A. At this locality, mares were kept with their foals in fenced paddocks all the time. The NaCl solution of d = 1.200 g/ml was generally the one that presented the highest frequency of diagnosis of nematode eggs and the highest mean of fecal eggs per gram. The fecal samples were also subjected to the polymerase chain reaction for amplification of DNA from the ITS2 region for Strongylus vulgaris. Twelve samples presented nucleotide sequences for S. vulgaris. In the end, this study revealed the high frequency (96.3%) of S. vulgaris among ponies on farms in Teresópolis, Rio de Janeiro, Brazil.

Comparison of McMaster and Mini-FLOTAC techniques for the diagnosis of internal parasites in pigs.

This purpose of this study was to compare the efficiency of the McMaster and Mini-FLOTAC quantitative techniques in the investigation of helminths in feces of pigs. An analysis was made of 74 fecal samples from pigs raised on family farms located in Rio de Janeiro, Brazil. These were analyzed by the Mini-FLOTAC and McMaster techniques in a solution of 1,200g/mL NaCl. This investigation revealed a superiority in the frequency of all helminths detected by Mini-FLOTAC, including Ascaris suum, Trichuris suis, strongyles and Strongyloides ransomi. The Kappa index revealed substantial agreement in all comparisons made in relation to the frequency of positive samples. However, significant statistical differences in the comparison of EPGs between McMaster and Mini-FLOTAC were observed for all nematodes (p ≤0.05). Higher values of Pearson's linear correlation coefficient (r), between the techniques in relation to EPG were observed for A. suum and T. suis, differently from what was observed for strongyles and S. ransomi. Mini-FLOTAC proved to be a more satisfactory and reliable technique both for the diagnosis of parasites and for the determination of EPG in pig feces due to the larger size of its counting chambers, thus increasing the helminth egg recovery rates.

Socioeconomic profile, animal care, sanitary practices, and knowledge about parasites among owners of domestic dogs and cats treated in Rio de Janeiro city.

Urban canine and feline populations are expanding worldwide, a fact that can boost the transmission of zoonotic parasites. Thus, it is necessary understanding dog and cat owners' profile, as well as their perceptions about zoonoses, to develop preventive strategies based on the One Health approach. The aim of the current study is to compare the profile of dog and cat owners, whose domestic pets were treated at Jorge Vaitsman Veterinary Medicine Center, Rio de Janeiro, Brazil. Dog and cat owners, whose domestic pets were treated in the routine service of the aforementioned medical clinic, from August 2017 to November 2018, filled a structured questionnaire comprising closed questions in order to collect information about their socioeconomic profile, hygiene habits, animal care and knowledge about parasites. Comparative analysis was applied to data collected from dog and cat owners, based on using Fischer's Exact Test or Pearson's Chi-square. In total, 350 individuals - 244 dog owners and 106 cat owners - filled the questionnaire. The comparative analysis of socioeconomic profile, animal care, sanitary practices and knowledge about parasites did not show differences between dog and cat owners (p≥0.05). Animal vaccination and deworming practices were significantly more prevalent among dog owners than among cat owners (p≤0.05). Overall, respondents have shown lack of knowledge about parasitic zoonoses. Results in the current study have evidenced the need of mediating and promoting information about potentially zoonotic parasitic agents among domestic pet owners, as well as the key role played by veterinarians as mediators of primary prevention measures against these agents.

As populações urbanas de cães e gatos estão em expansão, o que pode facilitar a transmissão de parasitas zoonóticos. Nesse sentido, é necessário conhecer o perfil dos donos de cães e gatos e suas percepções sobre zoonoses para construir estratégias preventivas em uma abordagem em Saúde Única. Este estudo teve como objetivo comparar o perfil dos donos de cães e gatos atendidos no Centro de Medicina Veterinária Jorge Vaitsman, Rio de Janeiro, Brasil. Entre agosto de 2017 e novembro de 2018, proprietários de cães e gatos atendidos na rotina da clínica médica responderam a um questionário estruturado composto por questões fechadas relacionadas ao seu perfil socioeconômico, hábitos de higiene, cuidados com os animais e conhecimento sobre parasitas. A análise comparativa dos dados obtidos de proprietários de cães e gatos foi realizada por meio do Teste Exato de Fischer ou Qui-quadrado de Pearson. No total, 350 indivíduos responderam ao questionário, sendo 244 donos de cães e 106 donos de gatos. A análise comparativa do perfil socioeconômico, cuidados com os animais, práticas sanitárias e conhecimento sobre parasitos não mostrou diferenças entre proprietários de cães e gatos (p≥0,05). As práticas de vacinação e desparasitação dos animais foram significativamente mais frequentes entre os donos de cães do que entre os donos de gatos (p≤0,05). De maneira geral, foi possível evidenciar entre as respostas recuperadas um desconhecimento sobre zoonoses parasitárias. Os resultados obtidos evidenciam a necessidade de mediação e divulgação de informações sobre agentes parasitários potencialmente zoonóticos para os proprietários, destacando os veterinários, no papel de mediadores das medidas de prevenção primária contra esses agentes.

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.

Infection by Prosthernorchis elegans (Diesing, 1851) in captive Callithrix aurita (É. Geoffroy, 1812) and Leontopithecus rosalia (Linnaeus, 1766) from Rio de Janeiro, Brazil.

This study reports on infection by Prosthernorchis elegans of Callithrix aurita and Leonthopithecus rosalia through biometry on adults and by molecular biology. Seventy-eight helminths were recovered from the animals' intestine. This is a detailed morphological description and the first molecular characterization of P. elegans in animals from Brazil.

Cryptosporidium species in non-human animal species in Latin America: Systematic review and meta-analysis.

Cryptosporidiosis is an infection caused by a protozoon that inhabits the gastrointestinal tract. More than forty valid species have been described in the genus Cryptosporidium, infecting a broad range of hosts around the world, some with zoonotic transmission and others with predominant anthroponotic transmission. Prevalence studies conducted in Latin American countries have been specific, without consolidating information on species prevalences. Thus, the aim of this study was to perform a systematic review and meta-analysis addressing the prevalence of Cryptosporidium species in animals in Latin America. The estimated pooled prevalence rate for cryptosporidiosis in animals, by means of meta-analysis with a random-effects model, based on species identification, was 18.0% (95% CI 11.0%-27.0%) with high heterogeneity. The estimated overall prevalence was 20.3% (36/177) in pets, 19.9% (1309/6573) in livestock animals and 23.9% (954/3995) in exotic/captive animals. Evidence of circulation of 16 Cryptosporidium species was found in five Latin American countries: Brazil, Colombia, Chile, Argentina and Mexico. Through meta-analysis with a random-effects model, the pooled prevalence rate for Cryptosporidium parvum was 0.7% (95% CI 0.2%-2.4%). Cryptosporidium felis (8.5%) was the most prevalent species in pets, C. parvum (10.3%) in livestock animals and Cryptosporidium galli (17.6%) in exotic/captive animals. C. parvum was the species with the greatest geographical dispersion, which can be explained by its eurixenic and zoonotic potential. Few studies on cryptosporidiosis in animals in Latin America were found, which shows that there is a need for investment in and expansion of studies on this parasite. The pooled prevalence of C. parvum in Latin America and its wide circulation are similar to what has been observed in other developing regions, which reaffirms the importance of this species as the cause of a neglected, emerging and zoonotic parasitosis.

Diagnosis, risk factors analysis and first molecular characterization of Cryptosporidium spp. in horses from Rio de Janeiro, Brazil.

An analysis was made of the frequency of Cryptosporidium spp. in fecal samples from horses raised on farms in the Teresópolis city, state of Rio de Janeiro, Brazil, and the risk factors that favored this infection. Between 2019 and 2020, 314 samples of equine feces were collected, 287 of which came from English Thoroughbred horses and 27 from ponies. Information on the horses and their management were retrieved from a stud book and forms filled out by trainers. The fecal samples were subjected to macroscopic analysis, modified Sheather's and Lutz parasitological techniques, safranin staining, and to enzyme-linked immunosorbent assay (ELISA) for the detection of coproantigens. All the samples that tested positive by these techniques underwent partial sequence analysis of the 18S rRNA gene to characterize the protozoan species. Cryptosporidium spp. was identified in 35 (11.1%) of the samples, 34 from English Thoroughbred horses and one from a pony. Based on a logistic regression model, it was found that the presence of dogs and small ruminants on the farms, and drinking water from a spring, were significantly associated with the animals' infection by the protozoan (p < 0.05). Eight of the English Thoroughbred horse samples underwent molecular characterization, which revealed the presence of Cryptosporidium felis in one sample and Cryptosporidium parvum in seven. The seven samples containing C. parvum were subjected to gp60 gene analysis, based on which nucleotide sequences typical of the IIa family were identified, which are usually transmitted from animals to humans. In addition, the genotype IIaA15G2R1, which is considered to have the highest profile of zoonotic transmissibility, was identified in one Thoroughbred horse. This is the first study conducted in the state of Rio de Janeiro that molecularly characterized Cryptosporidium spp. in horses, and the first on the American continent to detect C. felis in the feces of these animals.

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.

Balantidiasis in humans: A systematic review and meta-analysis.

Balantioides coli is a protozoan that infects different hosts species, including humans, with zoonotic transmission. The parasite, which lives in the large intestine and in other organs, can lead to serious infections that may culminate in death. Information about human balantidiasis is generally still very scanty. In view of the above, the purpose of this study was to analyze the epidemiological, clinical and laboratory characteristics of human balantidiasis based on a systematic review and meta-analysis. The scientific articles were retrieved from various databases and were subjected to descriptive analyses, chi-squared tests, and summarized on a forest plot and the heterogeneity index (I2). A total of 103 articles were eligible and included in this review. Out of these 103 articles, 75 were clinical case reports and 28 were epidemiological studies, indicating a frequency of 997 (3.98%) people potentially infected with B. coli. The publication dates of the analyzed articles ranged from 1910 to 2020, but the majority (68.9%) were published between 1998 and 2020. A considerable number of these articles were published in South America and Asia, mostly in Brazil and India, respectively. However, in Africa, Ethiopia, was observed the higher number of infected people (47.5%). A significant association (p < 0.05) was identified between proximity to pigs and positivity for B. coli, since more than 16% infected people were in proximity with pigs and/or their excreta. Infection by the protozoan was classified mainly as intestinal, and the predominant symptom was dysentery. Extraintestinal infections were found in 27 individuals, with colonization of the genitourinary tract frequently highlighted. Direct examination (17.2%), followed by an association of direct examination and sedimentation (45.7%), were the most commonly performed parasitological techniques, and the most frequently diagnosed form was trophozoites, corresponding to 22.5% of cases. The most common treatment for parasitized individuals (11.8%) was an association of tetracycline drugs with nitroimidazole derivatives. The articles retrieved, mainly epidemiological ones, used in meta-analysis showed high heterogeneity (I2> 50%, p < 0.05), impairing the retrieval and comparison of results. Some articles were found to provide incomplete information, making it difficult to retrieve and analyze variables. However, this review enabled us to compile and restate factors that appear to be associated with cases of human balantidiasis.

Toxoplasma gondii in domiciled dogs and cats in urban areas of Brazil: risk factors and spatial distribution.

Toxoplasma gondii is a highly prevalent zoonotic parasite in Brazil capable of infecting mammals and birds. The increase in the urban populations of pets and the narrowing of the human-animal relationship can facilitate the transmission of important public health zoonoses, such as toxoplasmosis. This study aimed to evaluate the frequency and spatial distribution of T. gondii infection and its risk factors in domiciled dogs and cats attended at the Jorge Vaitsman Institute, Rio de Janeiro. Serum samples from 400 dogs and 272 cats were evaluated by an indirect fluorescent antibody test (IFAT) for IgG anti-T. gondii antibodies. Epidemiological questionnaires were used to interview the animals' owners to identify risk factors for infection. Of the total, 34% (136/400) of dogs and 8.1% (22/272) of cats had anti-T. gondii antibodies. Breed (OR: 2.10-95%, CI 1.27-3.46) was a risk factor for dogs, while sex (OR: 3.40-95%, CI 1.10-10.52) and homemade food consumption (OR: 8.49-95%, CI 2.48-29.05) were risk factors for cats. Offal consumption was considered a risk factor for both species evaluated (OR: 2.74-95%, CI 1.38-5.43 for dogs; OR: 7.66-95%, CI 1.24-47.29 for cats). The spatial analysis showed that T. gondii seropositive animals were widely distributed in the metropolitan region of Rio de Janeiro state, with a concentration observed mainly in the west and north zones of Rio de Janeiro city. The results emphasize the importance of adopting prophylactic measures to control T. gondii transmission in domiciled dogs and cats in Rio de Janeiro, contributing positively to public health.

TITLE: Toxoplasma gondii chez les chiens et les chats domiciliés dans des zones urbaines du Brésil : facteurs de risque et répartition spatiale. ABSTRACT: Toxoplasma gondii est un parasite zoonotique très répandu au Brésil, capable d'infecter les mammifères et les oiseaux. L'augmentation des populations urbaines d'animaux de compagnie et le rétrécissement de la relation homme-animal peuvent faciliter la transmission de zoonoses importantes pour la santé publique, telles que la toxoplasmose. Cette étude visait à évaluer la fréquence et la distribution spatiale de l'infection à T. gondii et ses facteurs de risque chez les chiens et les chats domiciliés qui ont fréquenté l'Institut Jorge Vaitsman de Rio de Janeiro. Des échantillons de sérum de 400 chiens et 272 chats ont été évalués par un test d'immunofluorescence indirecte (IFAT) pour les anticorps IgG anti-T. gondii. Des questionnaires épidémiologiques ont été appliqués aux propriétaires des animaux pour identifier les facteurs de risque d'infection. Sur le total, 34 % (136/400) des chiens et 8,1 % (22/272) des chats avaient des anticorps anti-T. gondii. La race (OR : 2,10-95 %, IC 1,27-3,46) était un facteur de risque pour les chiens, tandis que le sexe (OR : 3,40-95 %, IC 1,10-10,52) et la consommation d'aliments faits maison (OR : 8,49-95 % IC 2,48-29,05) l'étaient pour les chats. La consommation d'abats a été considérée comme un facteur de risque pour les deux espèces évaluées (OR : 2,74-95 %, IC 1,38-5,43 pour les chiens; OR : 7,66-95 %, IC 1,24-47,29 pour les chats). L'analyse spatiale a montré que les animaux séropositifs pour T. gondii étaient largement répartis dans la région métropolitaine de l'État de Rio de Janeiro, avec une concentration observée principalement dans les zones ouest et nord de la ville de Rio de Janeiro. Les résultats soulignent l'importance d'adopter des mesures prophylactiques pour contrôler la transmission de T. gondii chez les chiens et les chats domiciliés à Rio de Janeiro, contribuant positivement à la santé publique.

Toxoplasma gondii among captive wild mammals in zoos in Brazil and Cuba: seroprevalence and associated risk factors.

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.

Intestinal parasites and risk factors in dogs and cats from Rio de Janeiro, Brazil.

The urban populations of pets, mainly dogs and cats, have been growing in recent years and, consequently, human-animal relationships have become increasingly closer. However, pets are vulnerable to infection by various parasites, as well are their owners due to the zoonotic potential of these agents. This study aimed to assess the frequency and risk factors of intestinal parasites in dogs and cats attended at the Municipal Institute of Veterinary Medicine Jorge Vaitsman, Rio de Janeiro, Brazil, as well as georeferencing the parasitized animals. A total of 608 animals (400 dogs and 208 cats) were enrolled from August 2017 to November 2018. Fecal samples were obtained from the animal's rectum during clinical examination. In addition, the owners of the enrolled dogs and cats were asked to complete an epidemiological questionnaire with questions about the animals' general information, lifestyle, and handling. Fecal samples were subjected to sedimentation and centrifugal-flotation in sucrose solution techniques. The frequencies of intestinal parasites total in dogs and cats were 11.3% and 24.5%, respectively, with hookworms being the most detected parasites in dogs (8.3%) and Dipylidium caninum in cats (12.5%). Sex was the only variable that showed statistical differences in the canine population males being more often infected than female (p < 0.05), while street access was associated with parasitic infections in cats (p < 0.05). The results obtained in this study demonstrate the need to control parasitic infections in domiciled dogs and cats in different municipalities in the metropolitan region of the state of Rio de Janeiro, including residents in the northern, western and central areas.

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.

Molecular evaluation of piroplasms and hematological changes in canine blood stored in a clinical laboratory in Niterói, Rio de Janeiro.

Piroplasm species were analyzed by molecular tools in total 31 blood samples from positive dogs, previously checked by stained slides, stored until DNA extraction between 2016 to 2018 in the laboratory Clinical Analyzes in Niterói, Rio de Janeiro. The piroplasms were identified by PCR, targeting the 18S rRNA gene and sequencing. From the total number of samples only 24 (77.4%) were positive and show adequate nucleotide sequences for interpretation with identity between 93%-100% with Babesia vogeli in compared to the sequences isolated of infected dogs from other states in Brazil deposited on GenBank. Most of dogs infected with B. vogeli had anemia (62.5%) and thrombocytopenia (95.8%). The findings of this study are compatible with previous reports in the literature and highlight B. vogeli as the most incriminated species in canine piroplasmosis in Brazil, and thrombocytopenia the hematological alteration most frequently identified in this infection. It is important to note that this is the first study involving the molecular characterization of piroplasms in the metropolitan region of Rio de Janeiro, based on PCR followed by sequencing.

Oncicola venezuelensis (Marteau, 1977) (Acanthocephala: Oligacanthorhynchidae) in Puma concolor in Rio de Janeiro, Brazil.

Specimens of Oncicola venezuelensis (Marteau, 1977) were recovered from fragments of intestinal tissue of a female Puma concolar (Linn, 1771) found dead in Petrópolis, Rio de Janeiro in 2017. A total of 140 helminths were recovered. Five males and 5 females of the helminths were analyzed morphologically as well as 50 parasite eggs recovered in intestinal contents. Morphologically, these helminths were compatible with the genus Oncicola, because of the size and shape of the proboscis, the size and disposition of the lemnisci and the morphometry of the eggs, in which the external membrane of the shell was delicate and clear. From histopathology, the helminths were deeply embeded in the mucosa reaching up to the muscle layer. One specimen was also identified molecularly with universal primers that amplified the eukaryote region ITS1-5.8S-ITS2. The helminth showed 99% identity with the gene sequence of O. venezuelensis deposited in GenBank. It is important to emphasize, this parasite has been very little reported in the literature, which reinforces the importance of this report.

Gastrointestinal Parasites in Captive Animals at the Rio de Janeiro Zoo.

BACKGROUND: Gastrointestinal parasites may determine diarrhea, dysentery or even death in captive mammals. These animals tend to be more susceptible to parasitic infections due to confinement and stress. Purpose To increase the information about these etiological agents in captive animals in Brazil, the gastrointestinal parasites of the captive mammals of the Rio de Janeiro Zoo were investigated. METHODS: From 2016 to 2018, 180 fecal samples were collected from animals housed in the Rio de Janeiro Zoo: 63 from animals of the order Primates, 26 of Carnivora, 78 of Artiodactyla, 9 of Perissodactyla and 4 of the order Rheiformes. The feces were processed by direct examination and by the techniques of Faust et al., Sheather, Ritchie, Lutz, and smears were stained with safranin. Immunoenzymatic assays were also performed to investigate antigens of Giardia duodenalis, Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar. RESULTS: Parasite positivity was identified in 68.3% of the fecal samples, with a parasite positivity rate of 68.2% among primates, 65.3% among carnivores, 69.2% among artiodactyls, 33.3% among perissodactyls, and 100% among rheiformes. The most frequently detected parasite was Entamoeba histolytica/E. dispar antigens, which showed a statistically significant positivity rate (33.3%; p = 0.000), particularly in the feces of carnivores (30.7%) and artiodactyls (53.8%). A statistically significant positivity rate of Balantioides coli (11.1%; p = 0.001) was also detected in feces from nonhuman primates, tapirs, collared peccaries and rheas. The positivity of Cryptosporidium sp. antigens in feces of the orders Carnivora, Artiodactyla and Primates was also statistically significant (7.2%, p = 0.010). Oocysts compatible with Cryptosporidium spp. were detected in 6.3% from primates. The helminths most frequently detected were thin-shelled eggs of nematodes (17.7%, p = 0.000), nematode larvae (15.5%, p = 0.000) and Trichuris trichiura eggs (6.1%, p = 0.018). CONCLUSION: The positivity rate for gastrointestinal parasites demonstrates the need for a sanitation management program to be implemented in the zoo, including routine diagnostic parasitology tests followed by specific treatment for each parasitosis.

Comparison of Four Parasitological Techniques for Laboratory Diagnosis of Eggs from Spirometra spp. in Wild Mammal Fecal Samples.

PURPOSE: This study aimed to compare the performance of four different microscopic coproparasitological techniques in relation to egg recovering and the frequency of alterations in the eggs observed through each technique. METHODS: A total of 213 fecal samples from free-living carnivorous mammals were collected between 2017 and 2018 in Itatiaia National Park, RJ, Brazil. Faust and modified Sheather floatation techniques as well as Lutz and modified Ritchie sedimentation techniques were applied. RESULTS: The total positivity rate for Spirometra spp. eggs was 24.4%, and these were detected mainly through Lutz and modified Ritchie sedimentation techniques, with substantial agreement (kappa = 0.73; p = 0.00 < 0.05; McNemar p value = 1.0; Fisher's exact test p = 0.616). Faust and modified Sheather flotation techniques did not present good egg recovery, with frequencies of 6.6% and 7.5%, respectively. Eggs with morphological alterations were mostly observed through Faust (17.3%) and modified Sheather (13.5%). Both flotation techniques presented statistically significant frequencies of deformed eggs, in comparison with the sedimentation techniques (p = 0.00). Low frequencies of deformed eggs were observed when the samples were analyzed through modified Ritchie and Lutz sedimentation techniques. CONCLUSIONS: From these results, sedimentation techniques such as modified Ritchie and Lutz methods were more efficient for diagnosing the eggs of this helminth in fecal material from free-living carnivores and should always be used when analyzing fecal samples from hosts of different species.