Toxoplasma gondii: isolation of tachyzoites rhoptries and incorporation into Iscom

Rhoptries have been isolated from Toxoplasma gondii tachyzoites by subcellular fractionation in isopynic density sucrose gradient. Five bands were observed, and transmission electron microscopy of these indicated that rhoptries were in band 3. This band had a density of 1.17g/cm(3). Fraction 1 had membrane structures of the parasite. Fraction 2 contained membranes and mitochondria (Fig. 1 B). Fraction 4 had mostly conoid structure (Fig. 2B) and fraction 5 showed ghosts. The electrophoretic and Western blotting analysis of the fractions indicated the presence of a number of proteins. Iscoms were constructed from band 3, which contained the rhoptry structures. Iscom showed a only protein incorporated of 55 kDa. Isolation of the parasite organelles has got in this work is necessary to identification, characterization, and function elucidation of the organelle proteins. (C) 2004 Elsevier B.V. All rights reserved.

PREVALENCE of SALMONELLA SPP. ANTIBODIES TO TOXOPLASMA GONDII, and NEWCASTLE DISEASE VIRUS IN FERAL PIGEONS (COLUMBA LIVIA) IN THE CITY of JABOTICABAL, BRAZIL

The rock pigeon (Columba livia) may serve as a reservoir for several pathogenic agents that can be transmitted to poultry, wildlife, domesticated pets, and/or humans via excreta, secretions, or dust from feathers. In addition, ingestion of infected pigeons by wild and domestic animals can also transmit these pathogenic agents. The health status of 126 free-living pigeons in an urban area was evaluated by microbiologic culture for Salmonella and serologic testing for the presence of antibodies for Toxoplasma gondii and for Newcastle disease virus (NDV) from 120 and 109 pigeons, respectively. After drawing blood, the birds were euthanized, and fragments of the liver, spleen, lungs, and gonads, and feces were cultured for Salmonella spp. Salmonella spp. was isolated from 10 birds (7.94%), of which 8 were Salmonella typhimurium, one was Salmonella enterica subsp. enterica serotype 4,12 and one was Salmonella enterica subsp. enterica serotype 4,12,i. Six of 109 pigeons (5.50%) were positive for NDV antibodies when using the hemagglutination inhibition test. Toxoplasma gondii antibodies were detected by immunofluorescence in one of 120 sera tested (0.83%). The results indicate that feral rock pigeons were exposed to NDV and T gondii, although the exposure was low. In addition, these birds had Salmonella spp. and could disseminate this pathogen in the environment.

Toxoplasma gondii: Evidence for the transmission by semen in dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Occurrence of anti-Neospora caninum and anti-Toxoplasma gondii IgG antibodies in goats and sheep in western Maranhão, Brazil

A neosporose e a toxoplasmose são doenças parasitárias que podem causar problemas reprodutivos em caprinos e ovinos. O objetivo deste estudo foi determinar a ocorrência de anticorpos IgG anti-Neospora caninum e anti-Toxoplasma gondii em caprinos e ovinos dos municípios de Amarante do Maranhão e Buritirana, microrregião de Imperatriz, Oeste maranhense, Nordeste do Brasil, bem como avaliar fatores associados à infecção por esses agentes etiológicos. Amostras de sangue de 110 animais (46 caprinos e 64 ovinos), provenientes de cinco propriedades, foram coletadas, e a reação de imunofluorescência indireta utilizada para o diagnóstico sorológico. Das 46 amostras de caprinos, 17,39% (n = 8) apresentaram anticorpos anti-N. caninum e 4,35% (n = 2) anti-T. gondii, enquanto das 64 amostras de ovinos, 4,69% (n = 3) e 18,75% (n = 12) apresentaram anticorpos anti-N. caninum e anti-T. gondii, respectivamente. Não houve diferença significativa, considerando-se a presença de gato e/ou cão na propriedade e assistência veterinária para ambos os agentes estudados. Entretanto, suplementação alimentar e presença de animais com problemas reprodutivos diferiram significativamente (p < 0,05) em ovinos e caprinos, respectivamente. Os resultados do presente estudo demonstraram que caprinos e ovinos, da região Oeste do Maranhão são expostos aos coccídios N. caninum e T. gondii. Essa é a primeira evidência desses agentes em pequenos ruminantes nessa região.

Detection of anti-Toxoplasma gondii antibodies in experimentally and naturally infected non-human primates by Indirect Fluorescence Assay (IFA) and indirect ELISA

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7

Toxoplasma gondii is an intracellular obligate protozoan, which infects humans and warm-blooded animals. The aim of the present study was to clone the rop2, gra5 and gra7 genes from T. gondii RH strain and to produce recombinant proteins. The rop2, gra5 and gra7 gene fragments produced by polymerase chain reaction were cloned into the pET102/D-TOPO(R) vector which contains thioredoxin and polyhistidine tags at the C-and N-ends, respectively, and is expressed in Escherichia coli BL21(DE-3). The expression fusion proteins were found almost entirely in the insoluble form in the cell lysate. These recombinant proteins were purified with an Ni-NTA column. Concentrations of the recombinant antigens produced in the E. coli BL21-star ranged from 300 to 500 mu g/mL growth media, which was used to immunize rabbits. We observed an identity ranging from 96 to 97% when nucleotide sequences were compared to GenBank database sequences. Immunocharacterization of proteins was made by indirect immunofluorescence assay. These proteins will be used for serodiagnosis and vaccination.

Alterações cognitivas em ratos infectados com Toxoplasma gondii

Toxoplasma gondii is a protozoan parasite that induces behavioral changes in rodents. The aim of this study was to evaluate the effect of infection by T. gondii during the chronic phase in working memory and impulsivity in rodents as well as the effect of antipsychotics to reverse any behavioral changes resulting from infection. Female Wistar rats (n = 40) were infected with 25 cysts of the strain ME-49 T. gondii after 4 months the animals were subjected to behavioral tests: tolerance to delay gratification, in which the animal must choose between two rewards, a smaller and more immediate, but delayed and the test of spontaneous alternation, in which the animal must use spatial cues to remember previously visited arms. Antipsychotic drugs were intraperitoneally administered during the testing of the behavioral experiments, the antipsychotic is haloperidol (1.5 mg / kg) administered 60 min before the start of the session and the antipsychotic clozapine (2.5 mg / kg) 30 min before. Animals infected with the parasite did not show operating deficits of memory, and motor impairment did not develop, however motor impairment was observed only in animals treated with haloperidol. It was found that administration of clozapine and haloperidol increased the percentage of alternation in infected and control groups in task switching espontânea.Não no distinction between control animals and infected the test of tolerance to delay gratification in relation to the percentage of choices greatest reward, during the pre-training and training, in which there is a delay of 15 s to access the great reward, however it was observed that infected animals prefer the greatest reward, when there is a delay of 30 s when compared to control group. The administration of clozapine possible that infected animals chose the greatest reward in the delay of 30 seconds during the test. These data suggest that infected mice do not exhibit deficits in working memory and that clozapine has therapeutic efficacy in improving cognitive performance of mice infected

Participação do circuito dopaminérgico nas alterações do comportamento de medo inato de camundongos infectados pelo Toxoplasma gondii

The protozoan parasite Toxoplasma gondii transforms the innate aversion of rats for cat urine into a fatal attraction, that increases the likelihood of the parasite completing its life cycle in the cat s intestine. The neural circuits implicated in innate fear, anxiety, and learned fear all overlap considerably, raising the possibility, that T. gondii may disrupt all of these nonspecifically. In this study, we evaluated immunoreactivity for tyrosine hydroxylase (TH) in areas associated with innate fear of infected male swiss mice. The latent Toxoplasma infection converted the aversion of mice to feline odors into attraction. This loss of fear is remarkably specific, as demonstrated by Vyas et al (2007), because infection did not diminish learned fear, anxiety-like behavior, olfaction, or nonaversive learning. However, the neurochemical mechanism related to alterations in innate fear due to T. gondii infection remains poorly studied. 20 mice were inoculated with bradyzoites (25 cysts) from a Toxoplasma gondii (Me-49 strain). The brains were removed after 60 days, sectioned and processed for TH immunohistochemistry. The correlation between the amount of cysts per area and the densitometric analysis of neurotransmitter reactivity was low in the areas implicated in innate fear of infected animals, when comparated with noninfected controls

Efeito do tratamento com as drogas antipsicóticas haloperidol e clozapina sobre a infecção de toxoplasma gondii em cultura de células retinianas embrionárias

T. gondii is an obligate intracellular protozoan and the main cause of retinochoroiditis in humans. The aim of this study was to evaluate the effect of the antipsychotic drugs haloperidol and clozapine on the course of infection by T. gondii of cultured embryonic retinal cells. Embryo retinas of Gallus gallus domesticus (E12) were used for the preparation of mixed monolayer cultures of retinal cells. Cultures were maintained on plates of 96 and 24 wells by 37°C in DMEM medium supplemented with 5% fetal bovine serum for 2 days. After this period, cultures were simultaneously infected with tachyzoites of T. gondii and treated with the antipsychotics haloperidol and clozapine for 48 hours. Treatment effects were determined by both assessing cell viability with the MTT method and evaluating infection outcomes in slides stained with Giemsa. The treatment with haloperidol and clozapine cells infected with T. gondii resulted in higher viability of these cells, suggesting a possible prevention of neuronal degeneration induced by T. gondii. Additionally, intracellular replication of this protozoan in cells treated with haloperidol and clozapine were significantly reduced, possibly by modulation of the parasite s intracellular calcium concentration

Genotyping of Toxoplasma gondii strains isolated from dogs with neurological signs

From 111 dogs with neurological signs admitted in this research in a 22-month period, brain samples of 34 animals were inoculated in mice in order to isolate Toxoplasma gondii. From these 34 dogs, 9 strains of T. gondii were isolated and the genetic characterization performed by restriction analysis (RFLP) of the SAG-2 gene. RFLP analysis showed that four of them were classified as Type I, and five as Type Ill. The present report is the first description of genotyping of T. gondii isolated from brain samples of naturally infected dogs, in Brazil. (C) 2004 Elsevier B.V. All rights reserved.

Leptospira spp. and Toxoplasma gondii antibodies in vampire bats (Desmodus rotundus) in Botucatu region, SP, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Risk factors and presence of antibodies to Toxoplasma gondii in dogs from the coast of São Paulo State, Brazil

A toxoplasmose é causada por um protozoário parasita intracelular obrigatório, Toxoplasma gondii, e acomete vertebrados homeotérmicos incluindo animais de companhia e o homem. O cão apresenta importância epidemiológica por atuar como sentinela da infecção para o homem. O presente estudo objetivou determinar a ocorrência de anticorpos para T. gondii em 205 amostras de soro de cães do município de Ubatuba, SP, Brasil, pela reação de imunofluorescência indireta (RIFI), assim como os fatores de risco relacionados à infecção nos animais, como raça, idade, sexo, acesso a rua, ingestão de comida caseira, acesso a água não tratada e presença de roedores. 52/205 (25,4%) amostras foram positivas para toxoplasmose, com títulos variando de 16 a 256. Os resultados sorológicos apresentaram associação significativa (P<0,05) com consumo de comida caseira (45/118; 38,1%; IC95% 29,9%-47,2%) (OR=7,0; CI95% 3,0-16,6), e acesso a rua, em que aqueles que não tinham acesso a rua foram prevalentes (37/121; 30,6%; IC95% 23,1%-39,3%) (OR=0,5; CI95% 0,2-1,0). Estes resultados demonstram que a toxoplasmose na região está relacionada com problema de educação sanitária, principalmente quanto ao adequado cozimento dos alimentos, visto que a maioria dos animais positivos não apresentou associação significativa com presença de roedores ou consumo de água não tratada, porém os mesmos permaneciam em casa aos quais fora oferecida comida caseira. Portanto, a toxoplasmose consiste em um problema de saúde pública na região estudada, sendo necessárias medidas sanitárias para o controle da infecção, visto a estreita relação homem-cão e os fatores de risco presentes.

Frequency of Toxoplasma gondii antibodies in tufted capuchin monkeys (Cebus apella nigritus) from an ecological station in the State of São Paulo, Brazil

A toxoplasmose é uma das zoonoses mais difundidas no mundo, causada pelo Toxoplasma gondii, um protozoário parasita intracelular obrigatório. Uma alta porcentagem de animais apresenta anticorpos específicos causados por exposição prévia, levando a uma infecção crônica. Os felídeos são os hospedeiros definitivos e outros animais homeotérmicos, incluindo os primatas, são os hospedeiros intermediários. Este estudo objetivou determinar a prevalência da infecção por T. gondii em macacos-prego (Cebus apella nigritus) de vida livre da Estação Ecológica localizada na Mata de Santa Teresa, Ribeirão Preto, SP, Brasil. Anticorpos anti-T. gondii foram pesquisados pelo método de aglutinação direta modificada (MAT) em amostras de soro de 36 macacos-prego, utilizando-se o título oito como de corte. Dos animais estudados, 3/36 (8,33%; IC95% 3,0-21,9%) apresentaram anticorpos anti-T. gondii, todos com título 32. Nenhuma diferença significativa (P>0,05) foi observada com relação ao sexo (1/3 machos e 2/3 fêmeas), e à idade (1/3 jovens e 2/3 adultos). Assim, estes resultados demonstram alta prevalência de anticorpos anti-T. gondii em primatas no estado de São Paulo.

Prevalence of antibodies against Toxoplasma gondii among Brazilian white-eared opossums (Didelphis albiventris)

Considering that Toxoplasma gondii is a parasite of global importance which affects several animal species including humans, the current study aimed to investigate the prevalence of antibodies against T. gondii among 72 white-eared opossums (Didelphis albiventris) from Botucatu Municipality (22 degrees 53'S 48 degrees 26'W), São Paulo State, Brazil. The investigation was carried out from January 2008 to December 2009, when the animals had their blood samples collected and subjected to the modified agglutination test (MAT); 12 specimens had brain samples bioassayed in mice. Seroprevalence was 5.5% (n = 4) and bioassays were negative. Older animals had higher prevalence of antibodies against T. gondii. Opossums in closer contact with the urban environment are likely more exposed to T. gondii than animals from the sylvatic environment. (C) 2011 Elsevier B.V. All rights reserved.

Anticorpos contra Toxoplasma gondii em equinos com histórico de ataxia

Stelmann U.J.P., Silva R.C., Langoni H., Borges A.S. & Amorim R.M. [Antibodies against Toxoplasma gondii in horses with history of ataxia]. Anticorpos contra Toxoplasma gondii em equinos corn historic de ataxia. Revista Brasileira de Medicina Veterinaria, 33(4):200-202, 2011. Programa de Pos-Graduacao em Medicina Veterinaria, Departamento de Clinica Veterinaria, Faculdade de Medicina Veterinaria e Zootecnia, Universidade Estadual Paulista Julio de Mesquita Filho, Campus de Botucatu, Distrito de Rubiao Junior, s/n, Botucatu, SP 18618-970, Brasil. E-mail: stelmann.ppgctia@gmail.comThe frequency of antibodies in blood serum and cerebrospinal fluid against Toxoplasma gondii in horses with a history of ataxia from the state of São Paulo, Brazil was carried out. Modified agglutination test was used to determine antibodies against T gondii, considering as positive samples with titers >= 2. of the blood samples tested, only 8 of 23 (34.78%) were positive, while CSF samples were negative when used the same technique. According to the negative results obtained for CSF, we concluded that T gondii was not the etiological agent of the myeloencephalitis in studied horses with neurological manifestation of ataxia, while the blood serological results indicated a previously exposure to T gondii.