Three new species of Eimeria Schneider 1875 in the montane grass mouse, Akodon montensis (Rodentia: Cricetidae: Sigmodontinae), and redescription of Eimeria zygodontomyis Lainson and Shaw 1990 from southeastern Brazil.

We describe three new coccidian species of the genus Eimeria Schneider 1875 (Apicomplexa: Eimeriidae) and redescribe and report Eimeria zygodontomyis Lainson and Shaw, 1990 in the montane grass mouse, Akodon montensis Thomas, 1913 from the Serra dos Órgãos National Park in southeastern Brazil. Sporulated oocysts of Eimeria zygodontomyis are ellipsoidal to cylindrical with a 0.6 (0.5-0.8) µm thick very delicate bi-layered wall; length × width (n = 49) 18.3 × 12.5 (16-20 × 11-13) µm; length/width ratio of 1.4 (1.2-1.6); 1 polar granule occasionally present; micropyle, residuum both absent. Sporocysts are ellipsoidal; length × width 8.5 × 5.2 (8-11 × 5-6) µm; length/width ratio of 1.5 (1.3-1.7) µm; Stieda body is prominent; sub-Stieda body is absent; sporocyst residuum is compact. Sporulated oocysts of Eimeria montensis n. sp. are spheroidal to subspheroidal with a 1.2 (1.0-1.4) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 30) 16.3 × 12.5 (15-17 × 13-15) µm; length/width ratio of 1.3 (1.0-1.4); 1 polar granule present; micropyle, residuum both absent. Sporocysts are ellipsoidal; length × width 7.2 × 5.1 (6-8 × 4-6) µm; length/width ratio of 1.4 (1.2-1.6); Stieda body is present, sub-Stieda body is absent; sporocyst residuum consists of small, scattered granules. Sporulated oocysts of Eimeria uricanensis n. sp. are ovoidal to pyriform with a 1.4 ( 1.3-1.6) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 40) 26.6 × 18.6 (23-30 × 17-20) µm; length/width ratio of 1.4 (1.3-1.6); 1 polar granule present; micropyle, residuum both absent. Sporocysts are ellipsoidal, length × width 13.3 × 8.0 (10-16 × 7-9) µm; length/width ratio of 1.7 (1.5-1.9); Stieda body, sub-Stieda body both absent; sporocyst residuum consists of a cluster of granules, forming a spheroid mass. Sporulated oocysts of Eimeria parnasiensis n. sp. are subspheroidal to ellipsoidal with a 1.8 ( 1.3-2.4) µm thick bi-layered wall; outer layer lightly pitted; length × width (n = 54) 28.2 × 21.9 (26-32 × 19-28) µm; length/width ratio of 1.3 (1.2-1.4); 1 polar granule present; micropyle is absent; oocyst residuum is present and consists of a cluster of granules of varying thickness. Sporocysts are ovoidal, tapering towards the Stieda body; length × width 12.2 × 7.6 (10-13 × 6-9) µm; length/width ratio of 1.6 (1.4-1.9); Stieda body is present; sub-Stieda body is absent; sporocyst residuum is present and consists of an aggregate of thin granules.

Prevalence of Anti-Toxoplasma Gondii Antibodies in Ratites from Brazil.

The prevalence of anti-Toxoplasma gondii antibodies was investigated among 104 ratites: 68 rheas (Rhea americana), 16 emus (Dromaius novaehollandiae), and 20 ostriches (Struthio camelus) in 4 Brazilian states. The prevalence in rheas was 26.5% (18 of 68), and titers were 1:100 (n = 8), 1:200 (n = 1), 1:400 (n = 4), 1:800 (n = 4), and 1:1,600 (n = 1). In emus, the prevalence was 50% (8 of 16), and titers were 1:50 (n = 1) and 1:100 (n = 7). The ostriches were slaughtered for human consumption, and 80% (16 of 20) were seropositive with titers of 1:200 (n = 1), 1:400 (n = 9), and 1:800 (n = 6). Sera were tested with a modified agglutination test, and the results confirmed the distribution of the parasite in ratite species from Brazil. The data obtained in this study show that T. gondii is prevalent among ratites from Brazil, and therefore ratite meat should also be considered a potential source of human infection. This is the first report of T. gondii seroprevalence in emus.

Epidemiological Investigation of Toxoplasma gondii Infections in Commercial Sheep Flock in an Endemic Area for Ocular Toxoplasmosis in Southern Brazil.

INTRODUCTION: Toxoplasma gondii is a widely distributed parasite and of great importance to human and animal health. METHODS: The objective of this study was to assess the prevalence of T. gondii antibodies and risk factors associated with the infection in sheep in the Northwest region of the State of Rio Grande do Sul, Brazil; this region has a very high rate of human ocular toxoplasmosis. Ovine sera were tested by the modified agglutination test (cut-off 1:25). RESULTS: T. gondii antibodies were detected in 70.2% (224 of 319). According to the logistic regression, the most significant factors associated were age and cat access to food stock facility. CONCLUSION: Preventive measures are discussed to reduce the risk of transmission of this zoonosis.

The Neurotropic Parasite Toxoplasma gondii Induces Sustained Neuroinflammation with Microvascular Dysfunction in Infected Mice.

Toxoplasmosis is one of the leading parasitic diseases worldwide. Some data suggest that chronic acquired toxoplasmosis could be linked to behavioral alterations in humans. The parasite infects neurons, forming immunologically silent cysts. Cerebral microcirculation homeostasis is determinant to brain functions, and pathologic states can alter capillarity or blood perfusion, leading to neurodegeneration and cognitive deficits. Albino mice were infected with Toxoplasma gondii (ME49 strain) and analyzed after 10, 40, and 180 days. Infected mice presented decreased cerebral blood flow at 10 and 40 days post infection (dpi), which were restored at 180 dpi, as shown by laser speckle contrast imaging. Intravital microscopy demonstrated that infection led to significant capillary rarefaction, accompanied by neuroinflammation, with microglial activation and increased numbers of rolling and adherent leukocytes to the wall of cerebral capillaries. Acetylcholine-induced vasodilation was altered at all time points, and blood brain barrier permeability was evident in infected animals at 40 dpi. Infection reduced angiogenesis, with a decreased number of isolectin B4-stained blood vessels and a decrease in length and branching of laminin-stained capillaries. Sulfadiazine reduced parasite load and partially repaired microvascular damages. We conclude that T. gondii latent infection causes a harmful insult in the brain, promoting neuroinflammation and microcirculatory dysfunction in the brain, with decreased angiogenesis and can contribute to a neurodegenerative process.

A new species of Eimeria Schneider, 1875 from the Serra dos Órgãos National Park, Rio de Janeiro, Brazil, with notes on its endogenous development in the montane grass mouse, Akodon montensis Thomas, 1913 (Rodentia: Sigmodontinae).

A total of 53 specimens of the montane grass mouse, Akodon montensis Thomas, 1913 were collected in the Serra dos Órgãos National Park (SONP) in November 2014 and July 2015. The fecal material was analyzed, and a prevalence of 7.5% was recorded for a new coccidian species of the genus Eimeria Schneider, 1875, with part of its endogenous development recorded in the small intestine. The oocysts of a new coccidian species of genus Eimeria are ellipsoidal to subspherical. The wall is bi-layered, c. 1.5 µm (1.3-1.6 µm) thick, outer layer rough. Oocyst (n = 126) mean length is 25.3 µm (21.0-28.0 µm), with a width of 20.2 µm (17.0-22.0 µm) and mean length/width (L:W) ratio of 1.3 (1.2-1.4). Polar granule is present, with the oocyst residuum as a large spherical to subspherical globule. Sporocyst shape (n = 126) is ellipsoidal, with a mean length of 11.8 µm (9.3-14.4 µm), width of 7.9 µm (6.7-9.3 µm), and mean L:W ratio of 1.5 (1.4-1.7). Sporocysts with nipple-like Stieda body and sub-Stieda body are absent. A sporocyst residuum formed by several globules, usually along the sporocyst wall. This is the first record of Eimeria in the montane grass mouse from Brazil.

Characterization of the zoonotic potential of Toxoplasma gondii in horses from Rio de Janeiro State.

The aim of this study was to perform a survey on the prevalence of anti-Toxoplasma gondii antibodies in horses from Rio de Janeiro State, Brazil. From 2012 to 2013, a total of 624 blood samples were collected from horses from the eight regions comprising Rio de Janeiro State (Baixadas Litorâneas, Serrana, Norte Fluminense, Noroeste Fluminense, Centro-Sul, Metropolitana, Médio Paraíba, and Costa Verde). All sera samples were tested for anti-T. gondii antibodies by performing the modified agglutination test with a cut-off of 1:25. Positive serology for T. gondii was detected in 22.8% (142/624) of the horses studied. Seropositivity was detected in all regions sampled; furthermore, statistical significance was observed when all locations were compared at once. The Médio Paraíba region had the highest number of positive animals 54.76% (23/42) in the Bonferroni correction among Costa Verde, Centro Sul, Metropolitana, and Serrana (p<0.001). Seropositivity was 39.58% (38/96) in Norte Fluminense, which was the second most prevalent region. The results indicated that the T. gondii parasite is widely distributed in horses in Rio de Janeiro State and represents a risk to public and animal health. These findings emphasize the need to increase control and prevention of this disease in horses.

Toxoplasma gondii transmission by artificial insemination in sheep with experimentally contaminated frozen semen.

Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 107 tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep.

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis.

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.

Occurrence and risk factors associated to Toxoplasma gondii infection in sheep from Rio de Janeiro, Brazil.

Toxoplasmosis is an important cause of abortion in sheep and a zoonotic risk to humans, leading to significant hazards to health and to economic losses. This study examined the seroprevalence and associated risk factors for infection with Toxoplasma gondii in 379 sheep from 12 flocks in Rio de Janeiro state, Brazil. Using the modified agglutination test (MAT), 202 (53.3 %) of 379 were seropositive with titers of 1:25 in 65, 1:50 in 40, 1:100 in 23, 1:200 in 11, 1:400 in 36, 1:800 in 7, 1:1,600 in 1, and 1:3,200 or higher in 19 sheep. The most significant factors associated with T. gondii seropositivity in sheep were age, gender, and veterinary care. Finding of T. gondii antibodies in 97 of the 202 sheep in titers of 1:100 or higher is indicative of persistently infected animals. This high level of seropositivity requires urgent control measures to reduce impact on animal productivity and public health.

Compromised nutrition in gerbils infected by Cystoisospora felis detected through an animal performance analysis tool.

The impact of Cystoisospora felis infection on the nutritional efficiency of gerbils was studied. The variables weight gain and feed intake were measured during four weeks in 28 laboratory gerbils, of which 14 were inoculated with 3.5 × 10(5) sporulated oocysts of C. felis and the remaining 14 were controls. The animals from both groups were weighted, killed, eviscerated and had their carcasses and tissues weighted and compared. A modern tool designed for measuring nutritional performance of farm animals was applied. The results showed compromised nutritional efficiency of the infected animals within the first week after infection. The consequences of these results are discussed here, including the potential impact of infection on farm animals performance.

Compromised nutrition in gerbils infected by Cystoisospora felis detected through an animal performance analysis tool / Comprometimento nutricional em gerbis infectados por Cystoisospora felis detectado através de uma ferramenta de análise de desempenho animal

The impact of Cystoisospora felis infection on the nutritional efficiency of gerbils was studied. The variables weight gain and feed intake were measured during four weeks in 28 laboratory gerbils, of which 14 were inoculated with 3.5 × 10(5) sporulated oocysts of C. felis and the remaining 14 were controls. The animals from both groups were weighted, killed, eviscerated and had their carcasses and tissues weighted and compared. A modern tool designed for measuring nutritional performance of farm animals was applied. The results showed compromised nutritional efficiency of the infected animals within the first week after infection. The consequences of these results are discussed here, including the potential impact of infection on farm animals performance.

O impacto da infecção por Cystoisospora felis na eficiência nutricional de gerbis foi estudado. As variáveis ganho de peso e consumo de ração foram mensuradas durante quatro semanas em 28 gerbis de laboratório, dos quais 14 foram inoculados com 3,5 × 10(5) oocistos esporulados de C. felis e os 14 restantes serviram como controle. Os animais de ambos os grupos foram pesados, mortos, eviscerados e tiveram suas carcaças e tecidos pesados e comparados. Uma ferramenta moderna desenvolvida para mensurar o desempenho nutricional de animais de produção foi aplicada. Os resultados mostraram eficiência nutricional comprometida dos animais infectados na primeira semana após a infecção. As consequências destes resultados são discutidas aqui, incluindo o possível impacto de infecção no desempenho de animais de produção.

Anti-Toxoplasma gondii antibodies in cattle and pigs in a highly endemic area for human toxoplasmosis in Brazil.

Sera from 77 cattle and 61 pigs from herds of Campos dos Goytacazes and neighborhood, north of Rio de Janeiro state, Brazil, were analyzed for the presence of antibodies anti-Toxoplasma gondii via ELISA. The seropositivity was confirmed at 49.4% (38 of 77) for cattle and at 11.5% (7 of 61) for pigs. Pigs bred under free-ranging systems presented with 20.6% (7 of 34) seropositivity and no pigs bred under indoor systems were positive (n  =  27). The seropositivity verified for cattle and pigs confirmed the extensive distribution of the parasite in this region. In the face of these data, we discuss the potential risk for human infection by consumption of raw or undercooked meat in this geographic region of Brazil. This is the first report of seropositivity for anti-T. gondii antibodies in cattle in the north of Rio de Janeiro State, Brazil.

Molecular characterization of Cryptosporidium in Brazilian sheep.

Feces were collected from 125 sheep between January and December 2007, on ten farms in the State of Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium. Ninety samples were collected from lambs 2 to 6 months of age, and 35 were from sheep over 12 months of age. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) in two steps of the SSU rRNA. Two samples (1.6%) from the lambs were positive, and after sequencing were identified as Cryptosporidium ubiquitum. This species has been reported worldwide and it is considered a zoonotic pathogen since it has been found and in several animal species and humans. However, because of the low frequency of C. ubiquitum found, the risk for public health in this region may not be high.

[Toxoplasma gondii in brains of pigs commercialized at the Municipality of Campos dos Goytacazes in the State of Rio de Janeiro, Brazil]. / Toxoplasma gondii em encéfalos de suínos comercializados no município de Campos dos Goytacazes, Estado do Rio de Janeiro, Brasil.

Brain samples of 12 pigs commercialized for food consumption at the Municipality of Campos dos Goytacazes in the State of Rio de Janeiro were examined for the presence of Toxoplasma gondii. Brain sample of each pig was grinded and submitted to peptic digestion. One ml of the homogenate of each sample was inoculated in three albino mice, and this procedure was repeated 24 hours later. Inoculated mice were observed during six weeks. Dead mice or those that presented clinical signs suggestive of the disease, were examined for the presence of tachyzoites in peritoneal exudates or tissue cysts in their brains. The percentage of positives brains was 50%, which is a confirmation that pigs could be considered as a potential source of infection of T. gondii for humans at this Municipality.