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.

Correction: Frequency of co-seropositivities for certain pathogens and their relationship with clinical and histopathological changes and parasite load in dogs infected with Leishmania infantum.

[This corrects the article DOI: 10.1371/journal.pone.0247560.].

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.

Lessons from a Multilaboratorial Task Force for Diagnosis of a Fatal Toxoplasmosis Outbreak in Captive Primates in Brazil.

Toxoplasmosis is an important zoonotic disease caused by the parasite Toxoplasma gondii and is especially fatal for neotropical primates. In Brazil, the Ministry of Health is responsible for national epizootic surveillance, but some diseases are still neglected. Here, we present an integrated investigation of an outbreak that occurred during the first year of the COVID-19 pandemic among eleven neotropical primates housed at a primatology center in Brazil. After presenting non-specific clinical signs, all animals died within four days. A wide range of pathogens were evaluated, and we successfully identified T. gondii as the causative agent within four days after necropsies. The liver was the most affected organ, presenting hemorrhage and hepatocellular necrosis. Tachyzoites and bradyzoite cysts were observed in histological examinations and immunohistochemistry in different organs; in addition, parasitic DNA was detected through PCR in blood samples from all specimens evaluated. A high prevalence of Escherichia coli was also observed, indicating sepsis. This case highlights some of the obstacles faced by the current Brazilian surveillance system. A diagnosis was obtained through the integrated action of researchers since investigation for toxoplasmosis is currently absent in national guidelines. An interdisciplinary investigation could be a possible model for future epizootic investigations in animals.

Serological Evidence of Hepatitis E Virus Infection in Brazilian Equines.

Hepatitis E virus (HEV) infection has been demonstrated in various animal species; those recognized as potential zoonotic reservoirs pose a considerable risk to public health. In Brazil, HEV-3 is the only genotype identified in humans and swine nationwide, in a colony-breeding cynomolgus monkey and, recently, in bovines and capybara. There is no information regarding HEV exposure in the equine population in Brazil. This study aimed to investigate anti-HEV antibodies and viral RNA in serum samples from horses slaughtered for meat export and those bred for sport/reproduction purposes. We used a commercially available ELISA kit modified to detect species-specific anti-HEV, using an anti-horse IgG-peroxidase conjugate and evaluating different cutoff formulas and assay precision. Serum samples (n = 257) were tested for anti-HEV IgG and HEV RNA by nested RT-PCR and RT-qPCR. The overall anti-HEV seroprevalence was 26.5% (68/257) without the detection of HEV RNA. Most municipalities (53.3%) and farms (58.8%) had positive horses. Animals slaughtered for human consumption had higher risk of HEV exposure (45.5%) than those bred for sports or reproduction (6.4%) (p < 0.0001). The statistical analysis revealed sex and breeding system as possible risk-associated factors. The first serological evidence of HEV circulation in Brazilian equines reinforces the need for the surveillance of HEV host expansion in a one-health approach.

Toxoplasma gondii infection in pig intended for human consumption: seroprevalence, risk factors and influence of biosecurity measures.

A serologic and epidemiologic study was carried out in order to determinate herd and animal seroprevalence and associated factors for Toxoplasma gondii in commercial pigs from Espírito Santo state, Brazil. Blood samples were collected from 416 pigs from 55 producer farms in 27 municipalities. An indirect immunofluorescent assay (IFA) was performed to estimate the seroprevalence of T. gondii and identify the associated risk factors using a questionnaire. The T. gondii antibody prevalence rate in commercial swine herds was 15.4% (64/416) using a cutoff of 1:64. The seropositivity for T. gondii was related to the presence of cats, water origin and age of swine in the increase of seroprevalence, and the existence of internal isolation fences and use of composting chambers as protective factors. To the best of our knowledge, this is the first study to report anti- T. gondii antibodies in the serum of pigs in the state of Espírito Santo, Brazil. This finding is important to public health because seropositive pigs can harbor tissue cysts in their meat, thereby representing a zoonotic risk for consumers of raw or undercooked porcine meat or its products.

Influence of killer immunoglobulin-like receptors genes on the recurrence rate of ocular toxoplasmosis in Brazil.

BACKGROUND: Recurrence is a hallmark of ocular toxoplasmosis (OT), and conditions that influence its occurrence remain a challenge. Natural killer cells (NK) are effectors cells whose primary is cytotoxic function against many parasites, including Toxoplasma gondii. Among the NK cell receptors, immunoglobulin-like receptors (KIR) deserve attention due to their high polymorphism. OBJECTIVES: This study aimed to analyse the influence of KIR gene polymorphism in the course of OT infection and its association with recurrences after an active episode. METHODS: Ninety-six patients from the Ophthalmologic Clinic of the National Institute of Infectology Evandro Chagas were followed for up to five years. After DNA extraction, genotyping of the patients was performed by polymerase chain reaction sequence-specific oligonucleotide (PCR-SSO) utilising Luminex equipment for reading. During follow-up, 60.4% had a recurrence. FINDINGS: We identified 25 KIR genotypes and found a higher frequency of genotype 1 (31.7%) with worldwide distribution. We note that the KIR2DL2 inhibitor gene and the gene activator KIR2DS2 were more frequent in patients without recurrence. Additionally, we observed that individuals who carry these genes progressed recurrence episodes slowly compared to individuals who do not carry these genes. MAIN CONCLUSIONS: The KIR2DL2 and KIR2DS2 are associated as possible protection markers against ocular toxoplasmosis recurrence (OTR).

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.

Toxoplasma gondii infection in pig intended for human consumption: seroprevalence, risk factors and influence of biosecurity measures / Infecção por Toxoplasma gondii em suínos destinados ao consumo humano: soroprevalência, fatores de risco e influência de medidas de biosseguridade

Abstract A serologic and epidemiologic study was carried out in order to determinate herd and animal seroprevalence and associated factors for Toxoplasma gondii in commercial pigs from Espírito Santo state, Brazil. Blood samples were collected from 416 pigs from 55 producer farms in 27 municipalities. An indirect immunofluorescent assay (IFA) was performed to estimate the seroprevalence of T. gondii and identify the associated risk factors using a questionnaire. The T. gondii antibody prevalence rate in commercial swine herds was 15.4% (64/416) using a cutoff of 1:64. The seropositivity for T. gondii was related to the presence of cats, water origin and age of swine in the increase of seroprevalence, and the existence of internal isolation fences and use of composting chambers as protective factors. To the best of our knowledge, this is the first study to report anti- T. gondii antibodies in the serum of pigs in the state of Espírito Santo, Brazil. This finding is important to public health because seropositive pigs can harbor tissue cysts in their meat, thereby representing a zoonotic risk for consumers of raw or undercooked porcine meat or its products.

Resumo Um estudo sorológico e epidemiológico foi realizado com o objetivo de determinar a soroprevalência nas granjas e animais e também em fatores associados para Toxoplasma gondii em suínos comerciais do estado do Espírito Santo, Brasil. Foram coletadas amostras de sangue de 416 suínos de 55 granjas produtoras de 27 municípios. A reação de imunofluorescência indireta (RIFI) foi realizada com o uso de questionário, para estimar a soroprevalência do T. gondii e identificar os fatores de risco associados. A taxa de prevalência de anticorpos contra T. gondii, nos rebanhos comerciais de suínos, foi de 15,4% (64/416) com ponto de corte de 1:64. A soropositividade para T. gondii foi relacionada à presença de gatos, origem da água, a idade dos suínos no aumento da soroprevalência, a existência de cercas de isolamento interno e uso de câmaras de compostagem como fatores de proteção. De acordo com este estudo, ele é o primeiro a relatar anticorpos anti-T. gondii no soro de suínos no estado do Espírito Santo, Brasil. Este achado é importante para a saúde pública, pois suínos soropositivos podem abrigar cistos teciduais em suas carnes, representando um risco zoonótico para os consumidores de carne suína crua ou mal cozida ou seus produtos.

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.

Subacute multisystemic toxoplasmosis in a captive black-and-gold howler monkey (Alouatta caraya) indicates therapy challenging.

A 10-year-old black howler monkey presented with a 36-day subacute clinicopathological picture of fever, prostration, inappetence, intestinal hypomotility, and emaciation. Therapy was trimethoprim-sulfamethoxazole with streptomycin. The liver, lungs, lymph nodes, and spleen presented lesions. Toxoplasma gondii isolation and PCR determined the diagnosis, and indirect fluorescent antibody tests confirmed an increase in antibody titers.

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.

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.

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.

Frequency of co-seropositivities for certain pathogens and their relationship with clinical and histopathological changes and parasite load in dogs infected with Leishmania infantum.

In canine leishmaniosis caused by the protozoan Leishmania infantum, little is known about how co-infections with or co-seropositivities for other pathogens can influence aggravation of this disease. Therefore, the objectives of this study were to evaluate the frequency of co-infections with or co-seropositivities for certain pathogens in dogs seropositive for L. infantum and their relationship with clinical signs, histological changes and L. infantum load. Sixty-six L. infantum-seropositive dogs were submitted to clinical examination, collection of blood and bone marrow, culling, and necropsy. Antibodies against Anaplasma spp., Borrelia burgdorferi sensu lato, Ehrlichia spp. and Toxoplasma gondii and Dirofilaria immitis antigens were investigated in serum. Samples from different tissues were submitted to histopathology and immunohistochemistry for the detection of Leishmania spp. and T. gondii. Quantitative real-time PCR was used to assess the L. infantum load in spleen samples. For detection of Coxiella burnetii, conventional PCR and nested PCR were performed using bone marrow samples. All 66 dogs tested positive for L. infantum by qPCR and/or culture. Fifty dogs (76%) were co-seropositive for at least one pathogen: T. gondii (59%), Ehrlichia spp., (41%), and Anaplasma spp. (18%). Clinical signs were observed in 15 (94%) dogs monoinfected with L. infantum and in 45 (90%) dogs co-seropositive for certain pathogens. The L. infantum load in spleen and skin did not differ significantly between monoinfected and co-seropositive dogs. The number of inflammatory cells was higher in the spleen, lung and mammary gland of co-seropositive dogs and in the mitral valve of monoinfected dogs. These results suggest that dogs infected with L. infantum and co-seropositive for certain pathogens are common in the region studied. However, co-seropositivities for certain pathogens did not aggravate clinical signs or L. infantum load, although they were associated with a more intense inflammatory reaction in some organs.

Genetic and histopathological characterization of Toxoplasma gondii genotypes isolated from free-range chickens reared in the metropolitan region of Rio de Janeiro state, Brazil.

This study aimed to genetically characterize Toxoplasma gondii isolates obtained from free-range chickens reared in the metropolitan region of the state of Rio de Janeiro, Brazil, and to evaluate the morbidity and histological changes associated with these isolates in mice. A mouse bioassay was used to isolate T. gondii from a pool of tissue samples (brain, heart, and thigh muscles) collected from 163 chickens. The 36 isolates obtained were genetically characterized by restriction fragment polymorphism (PCR-RFLP) analysis of the SAG1, 5'-3'SAG2, aSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3 genomic regions. Seventeen atypical genotypes were identified and nine of them were reported for the first time. All identified genotypes caused clinical signs and histological changes in mice, with the majority being associated with high cumulative morbidity (65%) and severe or very severe histological changes (76%). The exclusive identification of atypical genotypes, with a predominance of new genotypes, indicates great genetic diversity of T. gondii in the region studied. In addition, the finding that all identified genotypes caused clinical signs and often severe histological changes in mice suggests potentially relevant virulence of these strains.