Seroprevalence of Toxoplasma gondii in free-living Amazon River dolphins (Inia geoffrensis) from central Amazon, Brazil.

Toxoplasma gondii is an important pathogen in aquatic mammals and its presence in these animals may indicate the water contamination of aquatic environment by oocysts. Serum samples from 95 free-living Amazon River dolphins (Inia geoffrensis) from the Mamirauá Sustainable Development Reserve (RDSM), Tefé, Amazonas, Central Amazon, Brazil were tested for T. gondii antibodies using the modified agglutination test (MAT). Antibodies (MAT ≥ 25) to T. gondii were found in 82 (86.3%) dolphins with titers of 1:25 in 24, 1:50 in 56, and 1:500 in 2. Results suggest a high level contamination of the aquatic environment of the home range of these animals.

Isolation of Toxoplasma gondii from goats from Brazil.

Goats are economically important in many countries, and little is known of caprine toxoplasmosis in Brazil. Antibodies to Toxoplasma gondii were assayed in the sera of 143 goats from 3 Brazilian states, using modified agglutination test (MAT titer > or = 1:25); 46 (32.2%) tested positive. Samples of brain, heart, diaphragm, and masseter of seropositive animals were pooled, digested in pepsin, and bioassayed in mice. Viable T. gondii specimens were isolated from tissue homogenates of 12 goats; the isolates were designated TgGtBr1-12. Ten of the 12 isolates killed 100% of infected mice, indicating that goats can harbor mouse-virulent T. gondii and, hence, can serve as a source of infection for humans.

Isolation of Toxoplasma Gondii from goats from Brazil

Goats are economically important in many countries, and little is known of caprine toxoplasmosis in Brazil. Antibodies to Toxoplasma gondii were assayed in the sera of 143 goats from 3 Brazilian states, using modified agglutination test (MAT titer > or = 1:25); 46 (32.2%) tested positive. Samples of brain, heart, diaphragm, and masseter of seropositive animals were pooled, digested in pepsin, and bioassayed in mice. Viable T. gondii specimens were isolated from tissue homogenates of 12 goats; the isolates were designated TgGtBr1-12. Ten of the 12 isolates killed 100% of infected mice, indicating that goats can harbor mouse-virulent T. gondii and, hence, can serve as a source of infection for humans

Occurrence of Neospora caninum antibodies in capybaras (Hydrochaeris hydrochaeris) from São Paulo State, Brazil.

Capybara (Hydrochaeris hydrochaeris) is a large rodent distributed throughout tropical America. Antibodies to Neospora caninum in 213 feral capybaras from 11 counties of the State of São Paulo, Brazil, were assessed using the indirect immunofluorescent antibody test (titer >or=1:25) and found in 20 (9.4%), with titers of 1:25 in 4, 1:50 in 7, and 1:100 in 9 animals. This is the first report of occurrence of N. caninum antibodies in capybaras.

Seroprevalence and isolation of Toxoplasma gondii from sheep from São Paulo state, Brazil.

Sheep are important in the epidemiology of Toxoplasma gondii infection but little is known of ovine toxoplasmosis in Brazil. Antibodies to T. gondii were assayed in sera of 495 sheep from 36 counties of São Paulo State, Brazil, using the modified agglutination test (MAT titer > or =1:25) and found in 120 (24.2%). Samples of brain, heart, and diaphragm of 82 seropositive sheep were pooled, digested in pepsin, and bioassayed in mice. Toxoplasma gondii was isolated from tissue homogenates of 16 sheep and the isolates were designated TgShBr1-16. Six of the 16 T. gondii isolates killed 100% of infected mice. Results indicate that asymptomatic sheep can harbor mouse-virulent T. gondii, and hence they can serve as a source of infection for humans.

Isolation and molecular detection of Neospora caninum from naturally infected sheep from Brazil.

Neospora caninum was isolated from a naturally infected sheep from Brazil by bioassay in dogs. Approximately 70g of brain from each of two 4-month-old sheep with indirect fluorescent antibodies (>or=1:50) to N. caninum was offered to a different IFAT negative dog (Sheep n. 302, IFAT 1:400-Dog 1 and Sheep n. 342, IFAT 1:50-Dog 2). Parasite DNA was detected in both sheep brains using a PCR targeting the Nc-5 gene of N. caninum. Shedding of Neospora-like oocysts was noticed only in Dog 1, from 10 days post-inoculation (PI) to 25 days PI (a total of approximately 27,600 oocysts). Seventy days after infection, Dog 1 was euthanized and brain/cerebellum and medulla were collected and submitted to molecular methods, as were the oocysts, to confirm the identity of the isolate. Serum samples collected weekly from both dogs from the infection to the end of the experimental period had no antibodies anti-N. caninum by IFAT (<1:50). Oocysts, brain/cerebellum and medulla specimens of Dog 1 proved positive by a PCR assay targeting the Nc-5 gene of N. caninum. In addition, the oocysts have the DNA amplified by a PCR based on primers directed to the common toxoplasmatiid ITS1 sequence. The PCR products of ITS1 were sequenced, confirming again the isolate as N. caninum. Oocysts were also orally inoculated in two Swiss white mice two Mongolian gerbils (Meriones ungulatus) and two large vesper mice (Calomys callosus) (10(3)oocysts/animal). The rodents were sacrificed 2 months PI, and fresh preparations of brains showed Neospora thick-walled cysts in gerbil brains, but molecular detection using the Nc-5 PCR assay revealed DNA parasite in gerbil and also C. callosus brains. This is the first report of isolation and sequencing of N. caninum from a Brazilian sheep and the first report of molecular detection of N. caninum from C. callosus.

Seroepidemiology of Toxoplasma gondii and Neospora caninum in dogs from the state of Paraíba, Northeast region of Brazil.

A cross-sectional study was conducted to determine the seroprevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies and to investigate the risk factors related to antibodies against T. gondii and N. caninum in dogs of the city of Campina Grande, state of Paraiba, Northeast region of Brazil. For this purpose, 286 blood samples were collected from dogs during the rabies vaccination campaign, in September 2003, and on this occasion questionnaires addressing epidemiological aspects of the infections were given to each dog owner. The sera were analyzed for anti-T. gondii and anti-N. caninum antibodies by indirect fluorescent antibody tests. Of the total of 286 dogs, 129 were positive for T. gondii (titer16) with a prevalence value of 45.1% (95% CI=39.24-51.07%). For N. caninum, 24 dogs were positive (titer50), with a prevalence value of 8.4% (95% CI=5.45-12.23%). Antibodies to T. gondii and N. caninum were found simultaneously in 14 dogs (4.9%; 95% CI=2.7-8.08%). For T. gondii infection, the risk factors associated with seroprevalence was the age of the animals, with dogs older than one year presenting higher values of odds ratio, and co-habitation of cats in the household. For N. caninum infection, dogs that have street contact had higher odds of seropositivity than dogs that remained exclusively in a domestic environment.

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in ovine from São Paulo State, Brazil.

Serum samples from 597 sheep from São Paulo State, in the southeastern region of Brazil, were tested to determine the prevalence of antibodies directed against Toxoplasma gondii (> or = 1:64) and Neospora caninum (> or = 1:50) using the indirect fluorescent antibody test (IFAT). The animals were divided into three groups based on their age: < or = 1 year, 1-4 years, and > or = 4 years. Antibodies to T. gondii were observed in 34.7% of the samples with titers ranging from 64 to 16,384 and IgG antibodies directed against N. caninum were observed in 9.2%, with titers ranging from 50 to 3200. Only 3.5% of the sheep were positive for both agents. All farms had at least one positive animal for T. gondii, and 26 of the 30 farms had at least one positive animal for N. caninum. An association between seroprevalence and age was observed for T. gondii (P = 0.001), but not to N. caninum (P = 0.343). It was not possible to associate seroprevalence to T. gondii and the presence of domestic or feral cats, since in all farms there was at least one positive sheep. There was no association between seropositivity to N. caninum and the presence of domestic (P = 1.000) and feral dogs (P = 0.550).

Prevalence of Toxoplasma gondii antibodies in sera of domestic cats from Guarulhos and São Paulo, Brazil.

Antibodies to Toxoplasma gondii were determined in serum samples of 502 domestic cats from Brazil by the modified agglutination test (MAT), using formalin-fixed whole tachyzoites and mercaptoethanol. Antibodies (MAT > or = 1:20) were found in 132 (26.3%) of 502 cats. With respect to origin, antibodies were found in 26.7% of 430 stray cats from São Paulo, 10% of 40 stray cats from Guarulhos, and 40.6% of 32 cats from a cat breeder in São Paulo. Antibody titers were: 1:20 in 10 cats, 1:25 in 40 cats, 1:50 in 73 cats, and > or = 1:500 in 9 cats. Exposure rates of T. gondii in cats from São Paulo, Brazil are similar to that in domestic cats in North America.

Biological and genetic characterisation of Toxoplasma gondii isolates from chickens (Gallus domesticus) from São Paulo, Brazil: unexpected findings.

In spite of a wide host range and a world wide distribution, Toxoplasma gondii has a low genetic diversity. Most isolates of T. gondii can be grouped into two to three lineages. Type I strains are considered highly virulent in outbred laboratory mice, and have been isolated predominantly from clinical cases of human toxoplasmosis whereas types II and III strains are considered avirulent for mice. In the present study, 17 of 25 of the T. gondii isolates obtained from asymptomatic chickens from rural areas surrounding São Paulo, Brazil were type I. Antibodies to T. gondii were measured in 82 chicken sera by the modified agglutination test using whole formalin-preserved tachyzoites and mercaptoethanol and titres of 1:10 or more were found in 32 chickens. Twenty-two isolates of T. gondii were obtained by bioassay in mice inoculated with brains and hearts of 29 seropositive (> or =1:40) chickens and three isolates were obtained from the faeces of cats fed tissues from 52 chickens with no or low levels (<1:40) of antibodies. In total, 25 isolates of T. gondii were obtained by bioassay of 82 chicken tissues into mice and cats. All type I isolates killed all infected mice within 4 weeks whereas type III isolates were less virulent to mice. There were no type II strains. Tissue cysts were found in mice infected with all 25 isolates and all nine type I isolates produced oocysts. Infected chickens were from localities that were 18-200 km apart, indicating no common source for T. gondii isolates. This is the first report of isolation of predominantly type I strains of T. gondii from a food animal. Epidemiological implications of these findings are discussed.