Anticorpos para Leptospira spp., Toxoplasma gondii e Neospora caninum em cães errantes albergados em canil privado

The serological profile of 300 mongrel dogs of various ages and gender were investigated. Animals were captured in the streets and afterwards directed to a private kennel in Avare city (SP) to search for leptospirosis, toxoplasmosis, and neosporosis. Blood samples were obtained from jugular or cephalic vein for the obtention of sera. The microscopic agglutination test (MAT) was used to leptospirosis. MAT detect the prevalence of 9.3%. The most frequent reactant serovars were Bratislava (35.7%), Cynopteri (17.9%), Autumnalis (14.3%), and Copenhageni (10.7%), besides 7.1% to others serovars: Icterohaemorrhagiae, Canicola, and Hardjo. The modified agglutination test used for the diagnosis of toxoplasmosis showed 26% of positive animals, with titers varying from 16 to 256, with 16 in 3.3%, 64 in 13.7%, and 256 in 9% of the samples. To canine neosporosis, it was used the indirect fluorescent antibody test, and two animals (0.7%) demonstrated antibodies with titers 25 and 100. The results show the participation of the animals in the epidemiological chain of the researched diseases.

Prevalence and risk factors for human toxoplasmosis in a rural community

Toxoplasma gondii infection may lead to important pathological questions, especially in rural areas, where several sources of infection exist. Therefore, it is important to determine risk factors in order to establish adequate prophylactic measures. The present study aimed to assess the prevalence and risk factors involved in human toxoplasmosis infection in a rural community, in Eldorado, Mato Grosso do Sul State, Brazil. This community was composed of 185 farms - with 671 inhabitants - from which 20 were randomly chosen. In these farms, blood samples were collected from rural workers, who also answered a risk factor questionnaire. Serum samples were analyzed by means of direct agglutination test for the detection of anti-Toxoplasma gondii antibodies. From 73 samples collected, 79.45% were positive. None of the studied variables was significantly associated with the prevalence of the infection. However, among the individuals who reported eyesight impairments, 94.4% had anti-T. gondii antibodies, compared with 74.0% who did not report eyesight changes (p = 0.0594). Moreover, most individuals in the study (68.20%) were older than 18 years and presented 84.44% positivity, compared with 66.67% of positive individuals younger than 18 years old. We were able to conclude that a high prevalence of antibodies did not imply significant associations with the risk factors studied.

Detection of Toxoplasma gondii DNA in the milk of naturally infected ewes

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

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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Toxoplasma gondii: Comparison of a rhoptry-ELISA with IFAT and MAT for antibody detection in sera of experimentally infected pigs

Indirect ELISA and IFAT have been reported to be more sensitive and specific than agglutination tests. However, MAT is cheaper, easier than the others and does not need special equipment. The purpose of this study was to compare an enzyme linked immunosorbent assay using crude rhoptries of Toxoplasma gondii as coating wells (r-ELISA) with indirect fluorescence antibody test (IFAT) and modified agglutination test (MAT) to detect anti-T. gondii antibodies in sera of experimentally infected pigs. Ten mixed breed pigs between 6.5 and 7.5 weeks old were used. All pigs were negative for the presence of T. gondii antibodies by IFAT (titre < 16), r-ELISA (OD < 0.295) and MAT (titre < 16). Animals received 7 × 107 viable tachyzoites of the RH strain by intramuscular (IM) route at day 0. Serum samples were collected at days -6, 0, 7, 14, 21, 28, 35, 42, 50, and 57. IFAT detected anti-T. gondii antibodies earlier than r-ELISA and MAT. The average of antibody levels was higher at day 35 in IFAT (Log10 = 2.9) and in MAT (Log10 = 3.5), and at day 42 in r-ELISA (OD = 0.797). The antibody levels remained high through the 57th day after inoculation in MAT, and there was a decrease tendency in r-ELISA and IFAT. IFAT was used as gold standard and r-ELISA demonstrated a higher prevalence (73.3%), sensitivity (94.3%), negative predictive value (83.3%), and accuracy (95.6%) than MAT. Kappa agreements among tests were calculated, and the best results were shown by r-ELISA × IFAT (κ = 0.88, p < 0.001). Cross-reaction with Sarcocystis miescheriana was investigated in r-ELISA and OD mean was 0.163 ± 0.035 (n = 65). Additionally, none of the animals inoculated with Sarcocystis reacted positively in r-ELISA. Our results indicate that r-ELISA could be a good method for serological detection of T. gondii infection in pigs. © 2005 Elsevier Inc. All rights reserved.

Detection of enterotoxin and toxic shock syndrome toxin 1 genes in Staphylococcus, with emphasis on coagulase-negative staphylococci

The detection of staphylococcal enterotoxins is decisive for the confirmation of an outbreak and for the determination of the enterotoxigenicity of strains. Since the recognition of their antigenicity, a large number of serological methods for the detection of enterotoxins in food and culture media have been proposed. Since immunological methods require detectable amounts of toxin, molecular biology techniques represent important tools in the microbiology laboratory. In the present study, polymerase chain reaction (PCR) was used to identify genes responsible for the production of enterotoxins and toxic shock syndrome toxin 1 (TSST-1) in S. aureus and coagulase-negative staphylococci (CNS) isolated from patients and the results were compared with those obtained by the reverse passive latex agglutination (RPLA) assay. PCR detection of toxin genes revealed a higher percentage of toxigenic S. aureus strains (46.7%) than the RPLA method (38.3%). Analysis of the toxigenic profile of CNS strains showed that 26.7% of the isolates produced some type of toxin, and one or more toxin-specific genes were detected in 40% of the isolates. These results suggests the need for further studies in order to better characterize the pathogenic potential of CNS and indicate that attention should be paid to the toxigenic capacity of this group of microorganisms.

Utilização do método de aglutina ção direta e da reação de imunofluorescência indireta na detecção de anticorpos para Toxoplasma gondii em cavalos naturalmente infectados

Infection by Toxoplasma gondii in equines is usually not apparent, it being characterized by presence of antibody titers and tissue cysts. This study was aimed at verifying the presence of anti-Toxoplasma antibodies in equine serum by modified agglutination test (MAT) and reaction to indirect immunofluorescent antibody test (IFAT). 1984 samples of serum were examined, by MAT, using whole formalin fixed tachyzoites of T.gondii as antigen. The samples reacting in the MAT test, and 150 other negative samples, chosen at random, were also tested by IFAT, utilizing anti-equine IgG. The association among the test results was verified by the McNemar test. 138 samples were positive in the MAT test, with 60 (46.38%) presenting reaction at a dilution of 1:64; 52 (37.7%) at 1:256; 19 (13.8%) at 1:1024; five (3.6%) at 1:4096; and two (1.45%) at 1:16384. Of 132 positive MAT samples, 14 were negative in the IFAT test, but the statistical analysis indicated general agreement in results of the tests. The results obtained showed agreement among the tests utilized, and the possibility of participation of equines in the transmission of toxoplasmosis to carnivorous animals, and also to humans.

Evaluation of IFA, MAT, ELISAs and immunoblotting for the detection of anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs

The study evaluated the efficiency of diagnostic laboratory methods to detect anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs. 18-mixed breed pigs were used during the experiment; these were divided into two groups, G1 (infected group, n = 10) and G2 (uninfected group, n = 8). Infection was performed with 4 × 10 4 VEG strain oocysts at day 0 by the oral route in G1 animals. All pigs were euthanized at day 60, when retina, aqueous humour, and blood samples were collected. Anti-T. gondii antibody levels were assessed in serum (s) and aqueous humour (ah) by indirect immunofluorescence assay (IFA), modified agglutination test (MAT), m-ELISA (using crude membranes from T. gondii tachyzoites as antigen) and r-ELISA (using rhoptries from T. gondii tachyzoites as antigen). Polymerase chain reactions (PCR) of samples from the retina were performed by using Tox4 and Tox5 primers. Antibody titers of G1 animals ranged from 128 to 1024 and from 16 to 256 in serum and aqueous humour, respectively. There were differences in the correlation coefficients between IFA(s) × IFA (ah) (r = 0.62, P = 0.05), MAT(s) × MAT (ah) (r = 0.97, P < 0.0001); however, there was no significant difference between r-ELISA(s) × r-ELISA (ah) (r = 0.14, P = 0.7). Antibodies present in serum and aqueous humour recognized similar antigens. Samples of retina were positive by PCR in 30% (3/10) of infected pigs. G2 animals remained without antibody levels and were PCR negative throughout the experiment. These results suggest that the use of a combination of tests and immunoblotting for paired aqueous humour and serum samples could improve the sensitivity and specificity for the diagnosis of ocular toxoplasmosis. © 2007 Elsevier Ltd. All rights reserved.

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 ≥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. © 2009 American Society of Parasitologists.

Anticorpos anti-Leptospira spp. em animais selvagens do zoológico municipal de Ribeirão Preto, Estado de São Paulo, Brasil

Leptospirosis may affect all domestic and wild animals as well as human beings. Some serological studies have shown the involvement of wild species in the epidemiology of the disease. Once captive wild animals are not much studied, especially in Brazil, the present study aimed to detect anti- Leptospira spp. antibodies in animals from Ribeirão Preto city zoo, in São Paulo state, Brazil. Blood samples were collected from captive birds, fish, reptiles and mammals, sinanthropics and free-living animals and also from employees between March and October, 2006. Four hundred and three blood samples were obtained, 388 animals' samples (110 reptiles, 143 birds, 110 mammals and 25 fish) and 15 humans'. The sera were analysed by Microscopic Agglutination Test using 22 serovars from pathological leptospiras and two from non-pathological serovars. Among the animal samples, 339 were from captive animals, and 49 from free-living ones, captured with traps inside the zoo. One hundred and three (103/388 = 26.5%) samples reacted to leptospirosis, ninety-two (92/339 = 27.1%) samples were from captive animals and eleven (11/49 = 22.4%) from free-living ones. All humans' samples were negative. Serological titles varied from 40 to 5.120, with predominance of titles between 40 and 80 and the most frequent serovars were Patoc, Andamana, Canicola, Icterohaemorrhagiae and Panama.

Serological survey of Toxoplasma gondii in captive Neotropical felids from Southern Brazil

Toxoplasma gondii is the causative intracellular protozoan of toxoplasmosis in human being and animals. Members of the Felidae family are considered the single definitive host for the infection; both wild and domestic cats are able to excrete oocysts in the environment. Wild cats maintained in captivity may serve as source of infection for other clinically susceptible animals in the same environment. The aim of this study was to determine the frequency of T. gondii IgG antibodies in 57 neotropical felids (1 Leopardus geoffroyi; 3 Puma yagouaroundi; 17 Leopardus wiedii; 22 Leopardus tigrinus; and 14 Leopardus pardalis) kept at the Bela Vista Biological Sanctuary, Itaipu Binacional, Southern Brazil, by the modified agglutination test (MAT) using titer 16 as cut-off point. Seropositivity was observed in 38/57 (66.67%; 95% CI 53.66-77.51%) samples, with higher frequency in ocelots (71.43%). Wild-caught felids were three times more likely to be infected when compared to zoo-born animals (P≤ 0.05) and age of wild-caught animals (P= 0.6892; 95% CI. = 0.7528-1.66) was not significant as a risk factor for the infection, the same occurring with zoo-born animals (P= 0.05; 95% CI. = 0.6267-24.052). These results suggest that, despite efforts to control T. gondii infection in zoo facilities, such as individual pens, hygiene monitoring, veterinary care and pre-frozen meat offered as food, non-domestic felids kept in captivity, particularly the wild-caught specimens, may be invariably exposed to infection due to other environmental sources. © 2010 Elsevier B.V.

Falhas reprodutivas associadas com a presença de do vírus da artrite-encefalite caprina, toxoplasma gondii e neospora caninum em caprinos no estado de São Paulo, Brasil

This study aimed at assessing the occurrence of antibodies against the caprine arthritis-encephalitis virus (CAEV), Toxoplasma gondii and Neospora caninum, as well as the associations between the presence of antibodies and the occurrence of reproductive failures in goats. Serum samples were collected from 923 goats of both sexes, over 3 months of age, from 17 dairy farms located in different municipalities of São Paulo State, Brazil. Infections by T. gondii, N. caninum and CAEV were evaluated by indirect methods of diagnosis based on indirect fluorescence antibody test (IFAT), Neospora agglutination test (NAT), and agar gel immunodiffusion (AGID), respectively. A survey was conducted on the farms to obtain information about reproduction dates (abortions, stillbirths and births of weak and premature kids) and zoosanitary management. Antibodies against CAEV, T. gondii and N. caninum was found in 37.81%, 23.62% and 17.23% respectively. There was no significant association between the presence of anti-CAEV antibodies and CAEV/T. gondii or CAEV/N. caninum co-infection, suggesting that CAEV does not predispose goats to infection by these agents. However, when CAEV/T. gondii (p<0.01) or CAEV/N. caninum (p<0.001) co-infection was present, the occurrence of reproductive failures was significantly higher what could indicate that CAEV-induced immunosuppression may predispose goats to develop the clinical symptoms of toxoplasmosis and neosporosis increasing the risks of the reproductive failures.

Variáveis epidemiológicas e alterações clínicas, hematológicas e urinárias em cães sororreagentes para Leptospira spp

In order to determine antibodies anti-Leptospira spp. in dogs from Botucatu-SP with a clinical suspicion and the risk factors for this zoonosis, as well possible hematological and urine alterations were collected blood samples from 248 dogs. The diagnostic test performed was the Microscopic Agglutination Test (MAT) with 29 serovars of leptospires. Titers were considered reagent ≥200 and detected in 17.7% (44/248) of the dogs. The most frequent serovars were Autumnalis (20.5%); Pyrogenes (18.2%); Gryppothyphosa (15.9%); Canicola (13.6%); Bratislava and Copenhageni (9.1%); Andamana (4.5%) and Djasiman (2.3%). Regarding the analysis of the epidemiological questionnaire administered to owners of the dogs, there was statistical significance for sex, age and presence of rats at home, showing that, by logistic regression analysis, only the sex variable was been significantly associated to the presence of antibodies anti-Leptospira. The levels of urea and creatinine were significantly increased in the group of animals reagents to the Microscopic Agglutination Test, showing the renal damage in these animals and decreased hemoglobin level.

Inquérito sorológico, molecular e fatores de risco para leptospirose em mamíferos cativos, papel dos animais sinantrópicos presentes no local e aspectos de saúde pública

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