Alessandra Mara Alves Ragozo

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 ≥ 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 ≥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.

Genetic diversity among capybara (Hydrochaeris hydrochaeris) isolates of Toxoplasma gondii from Brazil

Recent studies indicate that Toxoplasma gondii isolates of many domestic hosts from Brazil are genetically and biologically different from T. gondii isolates from USA and Europe. However, little is known about genetics of T. gondii isolates from wild mammals in Brazil. In this study, genotypes of 36 T. gondii isolates from capybaras (Hydrochaeris hydrochaeris) from six counties in São Paulo state, Brazil, were determined. Sixteen genotypes were identified using 11 genetic markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. No classical clonal Type I and Type II isolates were found, confirming other findings that these lineages are rare in Brazil. Eight of these 36 isolates were grouped into the common clonal lineages in Brazil, previously designed as Types BrI, BrII and BrIII. Seven of the 16 genotypes were reported for the first time in this study. Three of the 36 isolates showed mixed infections. Analysis of mortality rates in infected mice indicated that Type BrI is highly virulent, Type BrII is intermediately virulent and Type BrIII is non-virulent, which is in agreement with previous report. The allele types at the CS3 locus are strongly linked to mouse-virulence of the parasite. These genotyping results support previous findings that the T. gondii population is highly diverse in Brazil.

Genetic diversity among Toxoplasma gondii isolates of small ruminants from Brazil: Novel genotypes revealed

Recent studies indicated that Toxoplasma gondii isolates of many domestic animal hosts from Brazil are genetically and biologically different from those in USA and Europe. Despite of high pathogenicity of this parasite to small ruminants, the epidemiology and genetic diversity of T. gondii in these animals are not well understood in Brazil. In this study, a total of 28 T. gondii samples (16 isolates from sheep in São Paulo state, and 12 isolates from goats in the states of São Paulo and Rio Grande do Norte) were genotyped using genetic markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. Eleven genotypes were identified from these T. gondii isolates. Eight isolates (4 from sheep and 4 from goats) were grouped into the common clonal type BrI lineage. One sheep isolate was grouped to the type BrIII lineage. Five isolates grouped to three previously identified genotypes in Brazil, and 13 isolates grouped to six novel genotypes. Mixed genotype was found in one isolate from goat in São Paulo. No classical clonal Type I, II or III isolates were found, confirming previous reports that these clonal lineages are rare in Brazil. The allele types at the CS3 locus are strongly linked to mouse virulence of the parasite. The results of this study indicate that even though a large number of T. gondii genotypes have been identified from a variety of animal hosts in Brazil, high percentage of new genotypes are continuously identified from different animal species, suggesting extremely high diversity of T. gondii in the population.

Ocorrência de parasitos gastrintestinais em fezes de gatos das cidades de Säo Paulo e Guarulhos

Fecal samples were collected from 138 cats with different sex and breeds captured from the streets of Sao Paulo and Guarulhos for the determination of gastrointestinal parasites infection. The animals were kept individually at Zoonosis Control Center at the cities of Sao Paulo (107 cats) and Guarulhos (31 cats). The feces were individually collected and examined by using sacarose solution flotation technique (d=1.203g/cm³). From the 138 samples, 80 (57.97%) were positive. Among the protozoa the most frequent agent was Cystoisospora felis in 36 cats (26.09%) followed by Cystoisospora rivolta in 34 cats (24.64%), Cryptosporidium parvum (1.45%) in two cats and Sarcocystis spp, in one cat (0.72%). Among the helminthes, Toxocara cati presented the high occurrence with 43 positive cats (31.16%), followed by Ancylostoma spp with 12 positive cats (8.70%) and Platynosomum fastosum in two cats (1.45%). Mixed infection were observed in 25 cats (18.12%) with T.cati and Cystoisospora spp. and T.cati and Ancylostoma spp, been the most common occurrence, both with 7.97% (11 samples) of occurrence.

Genotyping of Toxoplasma gondii isolates from free range chickens in the Pantanal area of Brazil

The aim of this paper was to genetically characterize Toxoplasma gondii isolates from free range chickens in regions of Brazilian territory in the state of Mato Grosso do Sul (MS) where T. gondii strains have never been studied. In total, T. gondii isolates from 22 free range chickens were included in this study. Fifty chickens from Eldorado, thirty from Rio Verde and ten from Aquidauana were sampled between January and April 2007. In relation to the genetic diversity of T. gondii isolates from chickens in MS, the magnitude of the diversity in the isolates sampled in this study was comparable to the overall diversity in a composite data set. These 22 isolates in MS revealed 11 genotypes, whereas the 321 isolates ever genotyped in Brazil have revealed 95 genotypes. The values of Simpsons Diversity Index for the whole population of T. gondii isolates in Brazil, the whole population of T. gondii isolates from chickens in Brazil and the population surveyed in this study were 0.97, 0.95 and 0.90, respectively. Seven of the 11 genotypes revealed from chicken isolates from MS are newly described genotypes and six of them each have a single isolate. In conclusion, the results obtained from isolates in MS corroborate previous studies on T. gondii isolates in Brazil, thus confirming their diversity and atypicality. Nonetheless, the applicability of PCR-RFLP markers for epidemiological inferences remains controversial.

ISOLATION OF TOXOPLASMA GONDII FROM CAPYBARAS (HYDROCHAERIS HYDROCHAERIS) FROM SAO PAULO STATE, BRAZIL

The capybara (Hydrochaeris hydrochaeris) is a large rodent used for human consumption in certain areas of South America. In the present study, viable Toxoplasma gondii was isolated for the first time from this host. Antibodies to T. gondii were assayed in the sera of 64 capybaras from 6 counties of São Paulo State, Brazil, using the modified agglutination test (MAT, ≥1:25 dilution) and the indirect fluorescent antibody test (IFAT, ≥1:16 dilution), and antibodies were found in 48 (75%) by MAT, and 49 (76.6%) by IFAT. Samples of brain, heart, and tongue of 40 seropositive capybaras were pooled, digested in pepsin, and bioassayed in mice. Toxoplasma gondii was isolated from tissue homogenates of 36 capybaras, and the isolates were designated TgCyBr1-36. Most isolates were lethal to mice; 17 of the 36 isolates killed 100% of infected mice, 11 isolates caused mortality in 25–90% of infected mice, and 8 isolates were nonpathogenic to mice. Results indicate that asymptomatic capybaras can harbor mouse-virulent T. gondii, and hence they 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 ≥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.

Serologic and molecular diagnostic and bioassay in mice for detection of Toxoplasma gondii in free ranges chickens from Pantanal of Mato Grosso do Sul

The aim of this study was to investigate the occurrence of Toxoplasma gondii and compare the results obtained in the Modified Agglutination Test (MAT), Polimerase Chain Reaction (PCR) and bioassay in mice. In order to accomplish this, 40 free-range chickens from eight farms in neighboring areas to the Pantanal in Nhecolândia, Mato Grosso do Sul, were euthanized and blood samples, brain and heart were collected. The occurrence of anti-T. gondii antibodies found in chickens was 67.5% (27 samples), considering as a cutoff point the dilution 1:5. Among the samples analyzed, 7 (25.9%) were positive in the dilution 1:5, 3 (11.1%) in 1:10, 2 (7.4%) in 1:20, 3 (11.1%) in 1:320, 1 ( 3.7%) in 1:640, 3 (11.1%) in 1:1280, 2 (7.4%) in 1:2560, 4 (14.8%) in 1:5120 and 2 (7.4%) in 1:10.240. From the mixture of tissue samples (brain and heart) from the chickens analyzed, 16 (40%) presented electrophoretic bands compatible with T. gondii by PCR (gene B1). In the comparison of techniques, 59.26% positivity in PCR was revealed among animals that were seropositive in MAT (cutoff 1:5). From 141 inoculated mice, six (4.44%) died of acute toxoplasmosis between 15 and 23 days after inoculation. Surviving mice were sacrificed at 74 days after inoculation, and a total of 28 cysts were found in the brains of 10 distinct groups. From the seropositive hens, 27 bioassays were performed and 11 (40.7%) isolates were obtained. A greater number of isolations happened in mice that were inoculated with tissues from chickens that had high titers for anti-T. gondii antibodies. Chronic infection in mice was observed in nine groups (33.3%) from five different properties. Among the surviving mice, 25.6% were positive for T. gondii in MAT (1:25). From mice positive in PCR, 87.5% were also positive in MAT. Among the PCR-negative mice, 5.2% were positive for T. gondii in MAT. It can be concluded through this study that the occurrence of infecton by T. gondii in the rural properties studied was high, that PCR directed to gene B1 does not confirm the viability of the parasite, but it can be used as a screening method for the selection of chickens infected by T. gondii, that the animals with titer greater than 10 must be prioritized for the selection of animals for bioassay, since for them, the chances of isolating the parasite are greater and that seroconversion in experimentally infected mice is not a good indicator for isolating the agent.

SEROLOGIC SURVEY OF INFECTIOUS DISEASES IN POPULATIONS OF MANED WOLF (CHRYSOCYON BRACHYURUS) AND CRAB-EATING FOX (CERDOCYON THOUS) FROM ÁGUAS EMENDADAS ECOLOGICAL STATION, BRAZIL

Abstract:  Domestic dogs are reservoirs for many infectious diseases and may represent a potential source of infection for wild canid populations. A serologic investigation of antibodies to Toxoplasma gondii, Neospora caninum, Brucella abortus, and Leptospira spp. was conducted on three maned wolves (Chrysocyon brachyurus) and seven crab-eating foxes (Cerdocyon thous), all free-living, at the Águas Emendadas Ecological Station (ESECAE), Federal District, Brazil, between February and October 2006. Out of the 10 samples analyzed, eight (80%) were seropositive for T. gondii: 3/3 (100%) of the maned wolves and 5/7 (71.4%) of the crab-eating foxes. None of the animals presented anti–N. caninum, B. abortus, and Leptospira spp. antibodies. This study demonstrated that the wild canid populations at ESECAE presented high exposure to T. gondii and indicated that there is high environmental contamination at the Station, which can be attributed to its proximity to urban zones, the presence of domestic cats in the study area, or the existence of other wild infected felines.

Serologic Survey for Selected Infectious Diseases in Free-Ranging Brazilian Tapirs (Tapirus terrestris) in the Cerrado of Central Brazil

Abstract From September 2000 to January 2002, a serologic survey was conducted in a population of free-ranging Brazilian tapirs (Tapirus terrestris) inhabiting Emas National Park and surrounding areas in Goiás state, central Brazil, as part of an ecologic study. Ten tapirs were immobilized with a tiletamine–zolazepam combination, and blood samples were collected. All sera were negative for Leptospira spp., Brucella abortus, and equine infectious anemia; and one of 10 animals was positive for Toxoplasma gondii. This report represents the first serologic survey for selected infectious diseases in a free-ranging population of Brazilians tapirs in central Brazil.