Immunomagnetic separation of Toxoplasma gondii and Hammondia spp. tissue cysts generated in cell culture.

Toxoplasma gondii is commonly transmitted among animals and humans by ingestion of infected animal tissues or by consumption of food and water contaminated with environmentally-resistant oocysts excreted by cats. Tissue cysts and oocysts have different walls, whose structures and compositions are poorly known. Herein, we describe an immunomagnetic separation (IMS) method that was successfully used for purification of T. gondii tissue cysts generated in cell culture. We used an IgG monoclonal antibody (mAb) that reacts against antigens in tissue cyst walls. Many in vitro produced cysts were obtained by this IMS; >2,000 T. gondii cysts were isolated from a single culture flask of 25 cm2. Tissue cysts from two Hammondia spp., H. hammondi, and H. heydorni, produced in cell culture were also separated using this method. As a reference, purification of tissue cysts by Percoll gradients was used. Percoll was able to separate T. gondii tissue cysts produced in mice but was not suitable for purifying T. gondii tissue cysts produced in vitro. The IMS described here should favor proteomic studies involving tissue cysts of T. gondii.

Genetic Diversity of Toxoplasma gondii Isolates From Free-Range Chickens In Bahia, Brazil.

The genotyping of 25 isolates of Toxoplasma gondii from free-range chickens in the state of Bahia, Brazil, was performed by PCR-restriction fragment length polymorphism using 11 genetic markers: SAG1, 5'+3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. The analysis revealed 8 genotypes, 3 of which had not been previously reported. Four genotypes were represented by single isolates, whereas the other genotypes were represented by 2 or more isolates. Five isolates showed mixed infections, and 2 of them were identical. None of the clonal types I, II, or III were found, but 2 isolates corresponded to the Brazilian clonal lineage BrIII. There was a single allele for the c22-8 marker. The CS3 marker demonstrated efficiency in the evaluation of virulence in mice. This study reaffirms the diverse genetic variability of T. gondii in Brazil.

In contrast to Toxoplasma gondii, Neospora caninum tachyzoites did not sustain multiplication in vitro at increased incubation temperatures.

Neospora caninum and Toxoplasma gondii are coccidian parasites that infect a wide range of mammalian and avian species. While viable T. gondii has been in vitro isolated in natural infections from wild and domestic birds, attempts to isolate N. caninum from naturally-infected birds were unsuccessful. We speculate that body temperatures of birds, which are usually higher than those of mammals, may impair the multiplication of N. caninum. In contrast to N. caninum, T. gondii can grow in vitro at temperatures higher than 37°C. To test the hypothesis that N. caninum tachyzoites are impaired to grow in vitro at high temperatures, three strains of N. caninum (NC-1, NC-Liverpool, and NC-Bahia) and three of T. gondii (RH, ME-49 and NED) were cultivated at gradually increasing temperatures starting at 37°C up to 41.5°C. A permanent chicken cell line was chosen for the study. Parasites were observed microscopically and their presence in culture was evaluated by species-specific conventional PCRs. In a second experiment, growth rates of T. gondii (RH strain) and N. caninum (NC-1 strain) were evaluated after direct passage of tachyzoites from 37°C to 41.5°C, and quantified by real-time PCR. In addition to comparisons between N. caninum and T. gondii, growth rates of three T. gondii strains were compared at high temperatures. Neospora caninum tachyzoites could not sustain multiplication at temperatures between 39°C and 41.5°C. Toxoplasma gondii tachyzoites continued to multiply at the same experimental conditions. Direct passage of N. caninum tachyzoites from 37°C to 41.5°C caused a significant decrease in the number of parasites during 96h of observation, while T. gondii had a significant increase in the number of stages after the same period of time. T. gondii RH strain (clonal type I) presented a different growth rate at 41.5°C when compared with type II and type III strains. In conclusion, multiplication of N. caninum tachyzoites in vitro was inhibited at temperatures similar to those of chickens, what may be one of the reasons that isolation of the parasite is difficult in avian species. In contrast to N. caninum, T. gondii continued to grow at 41.5°C.

Efeitos colaterais do uso da ribavirina, prednisona e DMSO em cães naturalmente infectados pelo vírus da cinomose / The side effects of ribavirin, prednisone and DMSO in dogs naturally infected by canine distemper virus

O estudo tem o objetivo de identificar efeitos indesejáveis da ribavirina, prednisona e DMSO em cães naturalmente infectados com o vírus da cinomose. Foram utilizados 60 cães apresentando quadro neurológico da cinomose com evolução de 10 dias. Os animais foram internados e receberam tratamento de suporte; foram avaliados diariamente e realizados hemograma, dosagem bioquímica e exame de urina tipo I. Os grupos 1 e 2 foram tratados com ribavirina e sua associação com DMSO; os grupos 3 e 4 com DMSO e prednisona e o grupos 5 com ribavirina e prednisona e o grupo 6 com ribavirina, prednisona e DMSO. Os animais foram anestesiados para a colheita de líquor, medula óssea e sangue, antes do tratamento para diagnóstico através da RT-PCR. As amostras negativas foram analisadas pela técnica de hn-PCR. Todos os animais apresentaram resultado positivo em pelo menos uma das duas reações. O efeito adverso da ribavirina e a sua associação com a prednisona foi a anemia hemolítica, que foi confirmada pela observação de bilirrubina na urina apenas dos cães tratados com ribavirina.

The present study aims at the identification of undesirable effects of ribavirin, predinisone and DMSO in dogs naturally infected by canine distemper virus. The research analyzed 60 dogs with clinical neurological signs and 10 days of evolution. The animals were hospitalized for the appropriate support treatment; were daily observed, and complete blood cells count, biochemical analysis, and urine exam type I were conducted. Groups 1 and 2 were treated with ribavirin and its combination with DMSO; Groups 3 and 4 treated with prednisone and DMSO, Group 5 treated with ribavirin and prednisone, while Group 6 with ribavirin, prednisone and DMSO. Before the treatment, animals were anesthetized for the cerebrospinal fluid, bone marrow and blood samples collection for the diagnosis based on RT-PCR. The negative samples were analyzed using the hn-PCR technique. All the animals presented positive results in at least one of the 2 tests. The adverse result of ribavirin and its association with prednisone was characterized by haemolytic anemia, confirmed by the evaluation of bilirrubin occurrence only in the urine of dogs treated with ribavirin.

A new set of primers directed to 18S rRNA gene for molecular identification of Cryptosporidium spp. and their performance in the detection and differentiation of oocysts shed by synanthropic rodents.

Cryptosporidium spp. are cosmopolitan protozoa that infect fishes, reptiles, amphibians, birds and mammals. More than 20 species are recognized within this genus. Rodents are a group of abundant and ubiquitous organisms that have been considered reservoirs of Cryptosporidium for humans and livestock. The aim of this study was to design specific primers for the gene encoding 18S rRNA, potentially capable of amplifying any species or genotype of Cryptosporidium spp. and evaluate the diagnostic attributes of the nested-PCR based on such probes. The primers were designed to amplify the shortest segment as possible to maximize the sensitivity of the test, but preserving the discriminatory potential of the amplified sequences for phylogenetic inferences. The nested-PCR standardized in this study (nPCR-SH) was compared in terms of sensitivity with another similar assay (nPCR-XIAO) that has been largely used for the detection and identification of Cryptosporidium spp. worldwide. We also aimed to molecularly characterize samples of Cryptosporidum spp. isolated from synanthropic rodents using these probes. Forty-five rodents were captured in urban areas of the municipality of Umuarama, Paraná State, Brazil. Fecal samples were submitted to three molecular tests (nested-PCRs), two of them targeted to the 18S rDNA gene (nPCR-SH and nPCR-XIAO) and the third targeted to the gene encoding actin (nPCR-actin). The nPCR-SH was tested positive on samples of Cryptosporidum parvum, Cryptosporidum andersoni, Cryptosporidum meleagridis, Cryptosporidum hominis, Cryptosporidum canis, and Cryptosporidum serpentis. Sixteen samples of rodents were positive by nPCR-SH, six by nPCR-XIAO and five by nPCR-actin. Sequencing of amplified fragments allowed the identification of Cryptosporidum muris in three samples of Rattus rattus, and two genotypes of Cryptosporidium, the genotypes mouse II and III. Cryptosporidium genotype mouse II was found in one sample of Mus musculus and genotype mouse III, in twelve samples, being five from R. rattus and seven from M. musculus. The results of this study demonstrated that the primers designed for detection of Cryptosporidium spp. were more efficient than those used in the nPCR-XIAO. Genotypes or species of Cryptosporidium that can be usually transmitted for human beings and livestock were not found in synanthropic rodents, suggesting that the importance of these animals in zoonotic transmission of cryptosporidiosis should be revisited.

Myenteric neuronal plasticity induced by Toxoplasma gondii (genotype III) on the duodenum of rats.

The effects of acute and chronic infection caused by Toxoplasma gondii on duodenal myenteric neurons were analyzed. Eighteen rats were assigned into four groups: Acute Control Group (ACG, n=4); Acute Experimental Group (AEG, n=4); Chronic Control Group (CCG, n=5); and Chronic Experimental Group (CEG, n=5). Rats from the AEG and CEG were inoculated orally with 105 genotype III (BTU-II strain) tachyzoites of T. gondii isolated from a dog with neurological signs. Acute groups were killed after 24 hours after the inoculation and the chronic groups after 30 days. Whole-mount from the duodenum were stained with Giemsa. The population density of myenteric neurons, as well the body cell, nuclear and cytoplasmic area were analyzed. Both acute and chronic toxoplasmic infection did not provoke neuronal loss. On the other hand, plastic alterations were observed: decreasing of the nuclear and cytoplasmic area during the acute phase and neuronal hypertrophy during the chronic phase.

Intraepithelial lymphocytes, goblet cells and VIP-IR submucosal neurons of jejunum rats infected with Toxoplasma gondii.

Toxoplasma gondii (T. gondii) crosses the intestinal barrier in oral infections and can lead to changes in different cell types, including the neurons located there. In the gastrointestinal system, the autonomous nervous system component that regulate blood flow and mucous secretion is the submucosal plexus. The aim of this study was to examine the effects of T. gondii infection on intraepithelial lymphocytes (IELs), goblet cells and submucosal neurons that are immunoreactive to vasoactive intestinal peptide (VIP-IR) of rat jejunum. Twenty male rats distributed as a control group (CG) and an infected group (IG), which received a suspension with 500 parasite oocysts (strain ME-49, genotype II) orally, were assessed. Routine histological sections were used to quantify IELs and to detect mucins secreted by goblet cells. Whole mounts including the submucosal layer were examined using immunofluorescence to detect the VIP neurotransmitter. Quantitative alterations in IELs were not observed. However, the reduction (P < 0.05) in the number of goblet cells that produce neutral mucins (PAS+) and sulphomucins (AB pH 1.0) and the maintenance of sialomucin-secreting cells (AB pH 2.5) resulting in a more fluid mucous were observed. Concerning the VIP-IR submucosal neurons, an increase in fluorescence on IG animals was observed. There was a reduction (P < 0.05) in the number of VIP-IR submucosal neurons and atrophy of their cell bodies in IG rats. Infection with T. gondii caused alterations in the chemical composition of the intestinal mucous and reduction in the neuron number and atrophy of the remaining neurons in this cell subpopulation.

Aspectos microbiológicos e de qualidade do leite bovino / Quality and microbiological aspects of bovine milk

A mastite é a principal afecção do gado leiteiro, possui alta prevalência, e constitui um fator limitante em muitas propriedades rurais do país, devido às perdas econômicas. Considerando-se a complexidade etiológica das mastites o objetivo do presente trabalho foi estudar os agentes de etiológicos desta enfermidade e a sua influência na qualidade do leite bovino. Para tanto, foram avaliados um total de 1090 tetos de animais de dez propriedades rurais localizadas no estado de São Paulo. A análise microbiológica do leite consistiu em cultivar uma alíquota de 0,1mL de leite de cada amostra positiva ao CMT, ou com mastite clínica, em meio de ágar base adicionado de 5 por cento de sangue ovino e em agar Mac Conkey, incubando-se as placas a 37°C com observação do desenvolvimento microbiano a cada 24 horas durante três dias. Os microrganismos com maior frequência na mastite foram Corynebacterium bovis(29,52 por cento), Streptococcus dysgalactiae (11,9 por cento) e Staphylococcus aureus (10,48 por cento). Houve ainda o isolamento em ágar Sabouraud dextrose de Candida krusei e Trichosporum spp. As médias de CCS e UFC dos animais foram variáveis e oito (80 por cento) propriedades encontram-se dentro dos limites estabelecidos para CCS pela Instrução Normativa n° 51 do Ministério da Agricultura, Pecuária e Abastecimento, e todas as propriedades se encontram dentro dos limites para UFC. Houve correlação positiva entre UFC e CCS de leite em duas propriedades entre as seis analisadas estatisticamente. Conclui-se que a mastite é um dos fatores que não permitem que o produtor atinja a qualidade exigida pelo governo. Falhas de manejo e higiene existem e devem ser corrigidas com treinamento dos produtores para aplicação de boas práticas de produção. Finalmente, o monitoramento das mastites e da qualidade do leite nos rebanhos deve ser realizado, e técnicas acessíveis como a CCS composta podem ser utilizadas.

Mastitis is the main disease in dairy herds, presents high prevalence and constitutes a limiting factor on many farms in Brazil due to economic losses. Considering the etiological complexity of mastitis the present work aimed to study the etiological agents of mastitis and its influence on the quality of bovine milk. For this, a total of 1090 teats from dairy cows of 10 farms localized in São Paulo state were evaluated. The microbiological analysis of milk consisted of cultivating an 0.1mL aliquot of milk from each CMT-positive sample, or with clinical mastitis, on agar base medium containing 5 percent of ovine blood and on McConkey agar, incubating plates at 37°C with observation of the microbial development at 24-hour intervals during three days. The microorganisms with highest frequency in mastitis were Corynebacterium bovis (29.52 percent), Streptococcus dysgalactiae (11.9 percent) and Staphylococcus aureus (10.48 percent). There was isolation of Candida krusei e Trichosporum spp. on Sabouraud dextrose agar. The averages of SCC and CFU from cows were variabl: eight (80 percent) farms were found to be within the limits established by regulation "Instrução Normativa nº 51" of the Ministry of Agriculture, and all farms were found to be within the limits for CFU. There was a positive correlation between CFU and SCC from milk in two of six farms statistically analyzed. It was concluded that mastitis is one of the factors that do not allow producer to reach the quality required by the government. Management and hygiene failures exist and must be corrected with instructions for the application of good production practices. Finally, monitoring of mastitis and milk quality in herds must be carried out, and accessible techniques as compound SCC can be used.

Isolation and multilocus genotyping of Toxoplasma gondii in seronegative rodents in Brazil.

Synanthropic rodents, mainly rats and mice, become ecologically associated with men due to changes in their ecosystems caused by human activities. These animals may take part in the epidemiological cycles of several diseases, including toxoplasmosis. The presence of serum antibodies to Toxoplasma gondii in 43 rodents captured in the urban area of Umuarama, PR, Brazil, was verified by modified agglutination test (MAT). Brain and heart samples were also collected and bioassayed in mice for the isolation of the parasite. Isolated samples were analyzed by 12 multilocus genotyping. Although all rodents were seronegative, the parasite was isolated in one mouse (Mus musculus) and one rat (Rattus rattus). Genotyping showed that these samples were similar to those previously isolated from cats in the state of Parana, Brazil.

Recrudescence of Toxoplasma gondii infection in chronically infected rats (Rattus norvegicus).

The kinetics of Toxoplasma gondii infection reactivation in the brain and muscles was analyzed in this study to determine the preferred tissue by the parasite during immunosuppression. Two groups of Wistar rats (G1 and G2) were inoculated with 10(4) bradyzoites of BTU10 strain (genotype I), p.o., and other two groups (G3 and G4) were inoculated with 0.9% saline solution. G2 and G4 were immunosuppressed with dexamethasone (DXM) and hydrocortisone sodium succinate (HSS). The presence of antibodies was researched in all groups through modified agglutination test (MAT) on days 0 and 21 p.i., and brain and muscle tissues of the rats were bioassayed in mice. G2 rats died at approximately 19.2days after drug treatment, while G1 rats survived. The reactivation was initially observed in G1 brain and G2 muscles. Thus, the initial reactivation in muscles after immunosuppression allows doctors to save precious time to control the evolution of reactivated infection, preventing brain damage to the host.

Efeitos da infecção crônica por Toxoplasma gondii sobre a parede intestinal de gatos domésticos / The effects of the infection caused by Toxoplasma gondii on the cat duodenal wall

O objetivo deste trabalho foi analisar os efeitos da infecção causada pelo Toxoplasma gondii sobre a parede do duodeno de gatos. Foram utilizados seis gatos (Felis catus), com cerca de três meses de vida, distribuídos aleatoriamente em Grupo controle (G1; n = 3) e Grupo infectado (G2; n = 3). Os animais do G2 receberam, por via oral, 200 cistos teciduais da cepa ME49 (tipo II) do T. gondii. Apos 40 dias, os animais foram submetidos a eutanásia, laparotomia e retirada do duodeno, que foi fixado em solução de Bouin e submetido a rotina histológica para obtenção de cortes transversais de3 μm. Os cortes foram corados com Hematoxilina-Eosina (HE), Azan, Acido Periódico de Schiff (PAS), Alcian-Blue e Tricrômio de Mallory. Realizou-se uma avaliação qualitativa da parede intestinal e medidas comparativas entre os dois grupos, com relação: à espessura da túnica mucosa, túnica muscular, parede total, altura dos vilos, profundidade das criptas e altura dos enterócitos e seus núcleos. As células caliciformes, os linfócitos intraepiteliais e as células de Paneth foram quantificados. Os resultados mostraram que a infecção levou a atrofia da túnica mucosa, túnica muscular e parede intestinal do duodeno de gatos do G2 (p < 0,05). A altura dos enterócitos apresentou um aumento significativo (p < 0,05) nos animais do G2. Na avaliação qualitativa, as fibras colágenas ocupavam visivelmente uma maior área dos estratos da parede intestinal, o que sugere que estejam aumentadas. Observou-se a redução da secreção de sulfomucinas e o aumento das células de Paneth nesses mesmos animais (p < 0,05).

This paper analyzes the effects of the infection caused by Toxoplasma gondii on the cat duodenal wall. Six cats (Felis catus) with 3-month-old were randomly divided into Control Group (G1; n = 3) and Infected Group (G2; n = 3). The animals from G2 received orarilly 200 T. gondii tissye cysts of ME49-strain (type II). After 40 days, the animals were submitted to euthanasia, laparotomy and had their duodenum removed, fixed in Bouin solution and submitted to histological routine obtaining 3 μm transverse cuts. The cuts were stained with Hematoxylin-Eosin (HE), Azan, Periodic acid – Schiff (PAS), Alcian-Blue, and Mallory trichrome. Qualitative assessment of the intestine wall as well as comparative measurements with respect to the thickness of mucosa, muscle tunic, total wall, the height of the villous, the depth of the crypts, and the height of the enterocytes and their nuclei were carried out. Calciform cells, the intraepithelial lymphocytes, and the Paneth cells were quantified. The results showed that the infection led to the atrophy of the mucosa, muscle tunic, and the intestinal wall of the duodenum of G2 cats (p < 0.05). The enterocytes height presented significant (p < 0.05) increase for G2 animals. According to the qualitative analysis, the collagen fibers were visibly taken a broader area on the intestinal wall layers, what suggests they have increased in size. Decrease in the sulphomucins secretion and the increase of Paneth cells were observed for these animals (p < 0.05).

Evaluation of Toxoplasma gondii placental transmission in BALB/c mice model.

Toxoplasma gondii infection is common worldwide and highly important to pregnant women as it can be transmitted to the fetus via the placenta. This study aimed at evaluating the prevention of placental transmission in two different strains after chronic infection with each one of the strains. A BALB/c mice model was inoculated 30days before breeding (immunization) and re-infected 12 and 15days after pregnancy (challenge). Seven experimental groups were assayed: G1: ME49-immunization (type II), M7741-challenge (type III); G2: M7741-immunization, ME49-challenge; G3, ME49-immunization; G4: M7741-immunization; G5: ME49-challenge; G6: M7741-challenge; G7: saline solution inoculation. Serology, mouse bioassay, PCR and RLFP of the uterus, placenta and fetus were performed to determine the congenital transmission of the strains challenged after chronic infection. IgG T. gondii antibodies were detected in G1, G2, G3 and G4, but not in G5, G6 and G7. All animals of G5 and G6 were IgM-positive. Congenital infection was not detected by bioassay and PCR. Nonetheless, placentas from G3 and G4 resulted positive but no corresponding fetal infection was detected. G1 and G2 did not show the genotype of the strain challenged during pregnancy, only those of chronic infection. Thus, the chronically infected BALB/c mice showed no re-infection after inoculation with another strain during pregnancy. Further studies with different parasite loads and different mice lineages are needed.

[Chronic infection due to Toxoplasma gondii inducing neuron hypertrophy of the myenteric plexus of Rattus norvegicus descending colon]. / Infecção crônica por Toxoplasma gondii induzindo hipertrofia de neurônios do plexo mientérico do colon descendente de Rattus norvegicus.

The effects of the chronic infection due to Toxoplasma gondii tachyzoites on the myenteric neurons of the adult rat descending colon were assessed in this study. Ten male, 60-day-old, Wistar rats, divided into control and experimental group were orally inoculated with 105 tachyzoites from Toxoplasma gondii genotype I strain. After 30 days, the animals were anesthetized and submitted to laparotomy. The descending colon was removed, dissected, and the whole-mounts were staining by Giemsa, in order to observe neurons of the myenteric plexus, followed by quantitative and morphometric analysis. It was verified that the infection caused alterations neither with respect to the dimensions of the organ nor the neuronal population; however, there was a significant increase of the perikarion area and the cytoplasm.

Infecção crônica por Toxoplasma gondii induzindo hipertrofia de neurônios do plexo mientérico do cólon descendente de Rattus norvegicus / Chronic infection due to Toxoplasma gondii inducing neuron hypertrophy of the myenteric plexus of Rattus norvegicus descending colon

Neste estudo, foram avaliados os efeitos da infecção crônica por taquizoítos de Toxoplasma gondii sobre os neurônios mientéricos do cólon descendente de ratos adultos. Utilizaram-se 10 ratos Wistar machos, com 60 dias de idade, divididos em grupo controle e experimental, que foram inoculados por via oral com 10(5) taquizoítos do genótipo I de T. gondii. Após 30 dias, os animais foram anestesiados e submetidos à laparatomia. O cólon descendente foi retirado, mensurado, dissecado e seus preparados de membrana submetidos à técnica de Giemsa, para coloração dos neurônios do plexo mientérico, seguido por análise morfométrica e quantitativa. Verificou-se que a infecção não causou alterações nas dimensões do órgão ou na população neuronal, porém houve um aumento significativo da área do pericário e citoplasma.

The effects of the chronic infection due to Toxoplasma gondii tachyzoites on the myenteric neurons of the adult rat descending colon were assessed in this study. Ten male, 60-day-old, Wistar rats, divided into control and experimental group were orally inoculated with 10(5) tachyzoites from Toxoplasma gondii genotype I strain. After 30 days, the animals were anesthetized and submitted to laparotomy. The descending colon was removed, dissected, and the whole-mounts were staining by Giemsa, in order to observe neurons of the myenteric plexus, followed by quantitative and morphometric analysis. It was verified that the infection caused alterations neither with respect to the dimensions of the organ nor the neuronal population; however, there was a significant increase of the perikarion area and the cytoplasm.

Virulence factors in Escherichia coli strains isolated from urinary tract infection and pyometra cases and from feces of healthy dogs.

The aim of this study was to compare the prevalence of virulence genes in 158 Escherichia coli strains isolated from 51 clinical cases of UTIs, 52 of pyometra and from 55 fecal samples from healthy dogs by PCR. papC was found in 12 (23.5%) strains isolated from UTIs, 19 (36.5%) from pyometra and 10 (18.2%) from feces. papGII was observed in 3 (5.8%) strains from pyometra, and papGIII in 10 (19.6%) from UTIs, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was detected in 22 (43.1%) strains from UTIs, 24 (46.1%) from pyometra and 19 (34.5%) from feces. hlyA was observed in 17 (33.3%) strains from UTIs, 18 (34.6%) from pyometra and 7 (12.7%) from feces, while cnf-1 was detected in 11 (21.6%) from UTIs, 21 (40.4%) from pyometra and 9 (16.4%) from feces. iucD was observed in 12 (23.5%) strains from UTIs, 9 (17.3%) from pyometra and 1 (1.8%) from feces. usp was found 17 (33.3%) isolates from UTIs and 36 (69.9%) from pyometra.

Comparação de métodos para determinação da prevalência de anticorpos anti-Toxoplasma gondii em suínos dos Estados de São Paulo e Pernambuco - Brasil / Methodologies Comparison for Prevalence of Anti-toxoplasma Gondii Antibodies Determination in Swine from São Paulo and Pernambuco States - Brazil / Comparacíon de metodologías para determinación de la prevalencia de anticuerpos anti-toxiplasma gondii en cerpos de los estados de São Paulo y pernambuco - Brasil

Pesquisou-se a presença de anticorpos anti-Toxoplasma gondii em soros de suínos, pelas técnicas de reação de imunofluorescência indireta (RIFI) e método de aglutinação direta (MAD). Foram examinadas 759 amostras de soros de suínos, oriundas de 25 propriedades, sendo 17 do Estado de São Paulo que responderam a questionário de levantamento de dados epidemiológicos e zoosanitários. O número de amostras positivas ao MAD foi 10 (1,32) e pela RIFI, 16 (2,11). Os resultados entre os testes apresentaram substancial concordância, com coeficiente kappa κ=0,6619, concordância de resultados positivos de 66,67 e concordância de resultados negativos de 99,46. Não foram encontradas associações importantes entre as variáveis epidemiológicas e zoosanitárias, e o resultado do exame sorológico.

ABSTRACT: There has been researched the presence of antibody against Toxoplasma gondii in swine sera by the techniques of immunofl uorescent antibody test (IFAT) and modifi ed agglutination test (MAT). There have been examined 759 samples of swine sera from 25 properties, being 17 in Sao Paulo state and 8 in Pernambuco. From São Paulo, 17 answered the questionnaire for the epidemiological and zoo sanitary study. The number of positive samples to the MAT was 10 (1.32%) and to IFAT 16 (2.11%). The results between the tests presented substantial agreement: kappa coeffi cient κ=0.66, agreement with positive results of 66.67% and agreement with negative results of 99.46%. There haven't been found important association between the epidemiological and zoo sanitary variables and the results of serological examination.

RESUMEN: Fue investigado la presencia de anticuerpos anti-Toxoplasma gondii en sueros de cerdos, por las técnicas de reacción de imunofl uorescencia indirecta (RIFI) y por el teste de aglutinación modifi cado (TAM). Fueron examinadas 759 muestras de 25 haciendas, 17 en el Estado de São Paulo y 8 en el Estado de Pernambuco. Siendo que de estas, 17 respondieron a un cuestionario para el estudio epidemiológico y zoosanitario. El número de muestras positivas en el TAM fue 10 (1,32%) y en el RIFI 16 (2,11%). Los resultados entre las pruebas presentaron una concordancia substancial: con coefi ciente kappa κ=0,66, concordancia de resultados positivos de 66,67% y concordancia de los resultados negativos de 99,46%. No fueron encontradas asociaciones importantes entre las variables epidemiológicas y zoosanitarias y el resultado del examen suerológico.