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This study aimed to research Toxoplasma gondii DNA in 102 samples of raw bovine milk from expansion tanks, in small properties located in different cities of the Midwest region of São Paulo, Brazil. For this, polymerase chain reaction (PCR) was performed with the primers TOX4/TOX5 for cPCR (conventional PCR), TgNP1/TgNP2 gene for nested PCR and the Tg18s58F/Tg18s348R for nested PCR. It was possible to detect T. gondii DNA in 18 (17.65 %) milk samples from the 102 tanks, corresponding to 4.90 % for TOX4/TOX5 primers, 12.74 % for TgNP1/TgNP2 gene and 0.98 % for Tg18s58F/Tg18s348R gene. The results showed that the TgNP1 and TgNP2 genes were more efficient to detect T. gondii DNA, and also indicated the importance of raw bovine milk as a source of human infections caused by this protozoan, being a public health problem. It is important to continue studies involving T. gondii from bovine milk considering the need for proper pasteurization, and for better comprehension regarding the epidemiology of this protozoan., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The objective was to describe the seroprevalence of anti-Neospora caninum antibodies in goats and sheep slaughtered in the state of Paraíba and to identify possible associated factors with the infections. Two hundred twenty-nine samples from goats and two hundred five from sheep were analyzed by Indirect Immunofluorescence Reaction (IFAT) using a cutoff point of 1:50. The presence of anti-N. caninum antibodies was identified in 28.4% (65/ 229; 95% Confidence Interval: 22.6-34.2) of the goat samples and in 12.7% (26/ 205; 95% CI: 8.2 - 17.2) of the sheep samples. Contact between goats and dogs (Odds ratio 4.81; CI 1.13 - 2.67; p = 0.041) and cattle (OR. 1.87; CI 1.13 - 2.67; p = 0.002) was identified as a risk factor for goats and contact between sheep and dogs (OR 2.32; CI 1.58 - 3.14; p = 0.026) and history of abortion (OR 1.94; CI 1.28 - 2.90; p = 0.001) was considered a risk factor for sheep. The study revealed a high seroprevalence of anti-N. caninum antibodies in slaughtered goats and sheep in Paraíba. Risk factors such as contact with dogs/cattle and abortion history underscore the need for preventive measures to control infection and enhance animal health management.

This study aimed to determine the occurrence of anti-Toxoplasma gondii, Neospora caninum, and Leptospira spp. antibodies in sheep and goats raised in villages of the Xukuru do Ororubá indigenous community, Pernambuco, Brazil. A total of 180 serum samples from sheep and 108 serum samples from goats of both sexes and different ages were analyzed. For antibody research, indirect immunofluorescence antibody test (IFAT) were used for the protozoa T. gondii and N. caninum, and microscopic agglutination test (MAT) for Leptospira spp., with a cutoff titer of 1:64, 1:50 and 1:100, respectively. The frequency of anti-T. gondii antibodies was 16.6% (30/180) for sheep and 11.1% (12/108) for goats. The frequency of anti-N. caninum antibodies was 10.55% (19/180) for sheep, and 20.37% (22/108) for goats, while for Leptospira spp., 2.2% (4/180) of sheep and 1.85% (2/108) of goats reacted positively. The results obtained in this study are unprecedented in indigenous communities in the country and serve as an alert for monitoring goats and sheep from the Xukuru do Ororubá indigenous village regarding the occurrence and productive impact of infections by T. gondii, N. caninum, and Leptospira spp., in addition to the occurrence of the zoonosis toxoplasmosis and leptospirosis in the indigenous community.

This study aimed to evaluate Toxoplasma gondii infection in pregnant goats. The goats were divided into two groups: group one (G1) comprised of 31 pregnant goats naturally infected with T. gondii, and group two (G2) comprised of seven uninfected pregnant goats from a flock with a history of abortion due to toxoplasmosis. Serological investigation, ultrasonography, and clinical testing were performed on all goats during gestation. Serum samples from goats and their offspring (precolostral) were collected to evaluate the vertical transmission of T. gondii. Samples from placentas and aborted fetuses were also collected for molecular and histopathological analysis. Results showed that in G1, estrus recurrence occurred in 22.6% (7/31) of the goats, embryonic death in 3.3% (1/31), and abortion in 19.4% (6/31). An increase in anti-T. gondii antibodies was observed in G1 goats at day 150 of pregnancy. T. gondii DNA was detected in 42.8% (3/7) of aborted fetuses and was associated with histopathological lesions caused by this parasite. Moreover, toxoplasmosis in field conditions caused by genotype ToxoDB #1 in pregnant goats resulted in severe reproductive loss in the flock., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Toxoplasmosis is an emerging and re-emerging infectious disease that can be transmitted through a contaminated environment. Environmental contamination is an emergency health issue, and determining its occurrence is fundamental to a One Health approach. In this study, we addressed the extent of environmental contamination and viability of Toxoplasma gondii oocysts in soil in different environments on Fernando de Noronha Island, Brazil. In addition, we performed species distribution modelling to predict the environmental suitability for coccidia persistence in the studied area. Soil samples were collected in 14 neighbourhoods of the Island and in the four main squares, creating a total of 95 soil samples (five samples per site). The samples were analyzed by the polymerase chain reaction (PCR) technique for the presence of the 18S ribosomal DNA gene of Apicomplexan protozoa, followed by genetic sequencing. We obtained 4.2% (4/95) positive soil samples with 100% similarity for T. gondii sequences. Two out of four positive sites on PCR showed viability of T. gondii oocysts through the mouse bioassay technique. As a result of the application of the species distribution modelling, environmental adequacy for the coccidia was observed throughout the Island. The results confirm the contamination of the soil in this insular environment by T. gondii oocysts and the environmental suitability by modelling application. These findings are an alert for the possibility of infection in animals and humans by contaminated soil, and for contamination of the maritime environment in addition to water resources for consumption by the local population., (© 2022 Wiley-VCH GmbH.)

This study aimed to describe the transmission of T. gondii in naturally infected goats in the semiarid region of northeastern Brazil, through evaluating the serological status of these animals from one to 12 months of age. Seven goat farms were selected. During the first visit, an epidemiological questionnaire was applied. Blood samples were collected from pregnant goats and from their female offspring every month, to monitor them for anti-T. gondii antibodies, using the indirect fluorescence antibody test (IFAT). Out of 66 pregnant goats evaluated, 47% (31/66) (95% CI: 34.6% - 59.7%) were positive and out of 93 female offspring, 61.3% (57/93) (95% CI: 52.4% - 69.2%) were positive, in at least one month during the evaluation period. Among the positive female offspring, 43.3% (27/57) (95% CI: 39.7% - 56.9%) presented environmental infection and 56.7% (30/57) (95% CI: 43.4% - 66.3 %) probably had only colostral antibodies or vertical transmission. It was observed that 92,6% (25/27) from environmental infections occurred until reproductive age (six months) (p<0.0001). It was also noted that there is a correlation between the titration of mothers and offspring, until four months of age (p<0.0001). Large fluctuations in the presence of antibodies were observed among the animals over the months monitored. It can be concluded that the prevalence of anti-T. gondii antibodies within the herds was greater than what has been expressed in specific prevalence studies and that most goat become infected up to the six months of life in the studied area., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

In this study, we determined the occurrence of Toxoplasma gondii oocysts in soil samples from public places. A total of 120 samples were collected from 24 sites, including squares, parks, university, hospitals in the city of Recife. The recovered oocysts were subjected to a nested-PCR test, and nine sites (9/24) were found to be positive for gene of apicomplexan parasites. The PCR product was sequenced, and 8.33% (10/120) of the samples showed 100% similarity to T. gondii DNA. T. gondii oocysts were detected in 75% (3/4) of the evaluated hospital soil samples and in 23.81% (5/21) soils samples from the public squares and parks. The results of this study demonstrate the potential of the soil in the areas analyzed as a source of T. gondii infection and therefore highlight the importance of devising educational strategies on the use of these sites, in addition to future cleaning protocols in public areas., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Toxoplasmosis, caused by the protozoan Toxoplasma gondii, is zoonotic disease and is one of the most important foodborne parasitic diseases globally. The prevalence in humans is highly variable, being influenced by cultural habits, socioeconomic, and environmental conditions. The objective of this study was to estimate the prevalence of T. gondii infection in humans on the archipelago of Fernando de Noronha, Pernambuco State, Brazil, and to identify the risk factors associated with this infection. The seroprevalence of immunoglobulin G anti-T. gondii antibodies was 50.4% (172/341, 95% CI: 45.2%-55.7%). Factors associated with the infection were consumption of well water or rainwater (odds ratio [OR]: 2.43, p=0.020) and consumption of game meat (OR: 1.80, p=0.026). This is the first study to provide epidemiological information of T. gondii infection among the residents of the Island of Fernando de Noronha, revealing a considerable antibody seroprevalence in this population. This study provides information for the adoption of prevention and control measures in island environments.

Little is known about Toxoplasma gondii infection among cattle living in the Cerrado (Brazilian savanna) biome in Brazil. In particular, there is no epidemiological data relating to infection in quilombo lands, i.e. areas settled by Afro-descendants of escaped slaves. The aim of this study was to determine the prevalence, spatial distribution and risk factors associated with T. gondii infection among cattle in the Kalunga quilombo, in the Cerrado biome. Blood samples were collected from 1533 cattle for antibody detection using the indirect fluorescence antibody test (IFAT). The study area was subdivided into five macroregions to determine the spatial distribution of infection. An objective questionnaire was applied to the cattle owners to evaluate risk factors, which were analyzed using univariate analysis and logistic regression. The prevalence of T. gondii infection among cattle was 8.93% (137/1533), and antibodies were found in 49.6% of the herds (66/133), in all macroregions. The risk factors associated with T. gondii infection in cattle were the following: number of animals in the herd (OR: 30.56), purchase of cattle (OR: 2.57), age group (OR: 1.95) and average annual temperature (OR: 1.77). Thus, the occurrence rate, spatial distribution and risk factors associated with T. gondii infection among cattle in the Kalunga quilombola community are documented here, for the first time.

Toxoplasmosis is a parasitic disease, which is prevalent across the world and has a strong environmental component in its transmission chain. The present study determined the seroprevalence and factors related to Toxoplasma gondii infection among free-range chickens from an Atlantic Forest area in Northeastern Brazil. A total of 550 free-range chickens were tested for T. gondii antibodies usingthe indirect fluorescent antibody test (IFAT, cut-off point 1:16), and 269 (48.9%; 95% CI = 44.7-53.1%) were positive. Among the 61 properties 57 (93.4%) had at least one seropositive animal. The robust Poisson regression model showed that the variables/categories associated with the seroprevalence of T. gondii were: region of origin of the animals/urban area (prevalence ratio [PR] = 2.346; P<0.001), management system/use of cages (PR = 1.591; P = 0.019), presence of rodents/yes (PR = 1.295; P = 0.035), and the type of food/use of food scraps (PR = 1.603; P = 0.009). The high prevalence found suggest that this scenario demands careful management, mainly regarding the use of scraps of undercooked or raw food, adequate environmental hygiene and frequent rodentcontrol.

Purpose: The aim of the present study was to detect and genotype T. gondii in free-range chickens destined to human consumption in Alagoas state, Brazil., Methods: Two hundred blood samples were collected from free-range chickens and submitted to indirect immunofluorescence antibody test (IFAT). Brain tissue from 14 animals randomly selected were subjected to mouse bioassay. Positive samples in mouse bioassay were submitted to PCR and genotyped by PCR-RFLP., Results: Out of two hundred blood samples from chickens, 72 (36%) samples were considered positive by IFAT. Two T. gondii strains were isolated, both being characterized as atypical and classified as #146 and a new genotype, named #279 in ToxoDB database., Conclusions: Our results showed a sero-occurrence of T. gondii in free-range chickens intended for humans, and the genetic diversity of the parasite in Brazil, with a new genotype described.

There is limited available information concerning the prevalence of Toxoplasma gondii in noncaptive monkeys. Also, New World monkeys (NWM) are highly susceptible to toxoplasmosis, which is a conservation concern. This study aimed to investigate apicomplexan parasites in common marmosets (Callithrix jacchus) collected for yellow fever and rabies surveillance program in Northeastern region of Brazil. Heart fragments of 39 free-ranging common marmosets were analyzed for the presence of the 18S rDNA gene of apicomplexan parasites by nested PCR. Positive samples were sequenced. T. gondii DNA was detected in 17.9% (7/39) of the analyzed animals. This study is the first report on T. gondii in Callithrix jacchus in Brazil. These findings should be an alert for wildlife conservation institutions, as high susceptibility and mortality were reported for captive NWM., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Beef cattle farming in the Amazon region has expanded rapidly, but information on herd health is still scanty. The purpose of this study was to determine the presence of anti-Toxoplasma gondii antibodies in cattle, its spatial distribution and the risk factors associated with the infection in the state of Amazonas, Brazil. Blood samples were collected from 1073 animals on 47 farms, located in 33 municipalities in the four state subpopulations. Anti-T. gondii IgG antibodies were detected by means of the indirect fluorescent antibody test (IFAT). The overall prevalence was 30.9 % (332/1073), and seropositive animals were identified at 93.6 % farms (44/47). All the subpopulations studied in the state of Amazonas had cattle herds seropositive for T. gondii, with some areas showing higher prevalence rates. The risk factors identified in the logistic regression were number of animals (OR = 4.43) and presence of domestic cats (OR = 1.98). It is advisable to correct identified risk factors, particularly insofar as the definitive hosts of T. gondii are concerned. Attention should also focus on beef consumption, given the prevalence of T. gondii infection among cattle and the widespread clandestine slaughtering that occurs in this state., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Toxoplasma gondii is a zoonotic parasite which can infect almost all warm-blooded animals. Toxoplasma gondii isolates from Brazil have greater genetic diversity with a predominance of virulent and atypical genotypes, compared with the Northern Hemisphere. Considering that previous studies have demonstrated a high seroprevalence of T. gondii antibodies in animals from Fernando de Noronha Island, the aim of this study was to isolate, genetically characterize, and determine mouse virulence of isolates of T. gondii from livestock from this Brazilian island. Two T. gondii isolates were obtained by mouse bioassay from brain from one sheep and one pig. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, PK1, L358, and Apico) and an atypical genotype of T. gondii (ToxoDB #146) was identified for both isolates. Genotyping of four ROP loci indicated different alleles for ROP16 and mouse virulence analysis revealed different profiles (intermediate and low virulence). This is the first report of this genotype being described in a pig and a sheep.

The ability to differentiate from the proliferative (tachyzoite) to the latent (bradyzoite) stage of isolates of Toxoplasma gondii recombinant genotypes (I/II/III and I/III) and reference strains from a clonal line (RH and ME49) was investigated in this study. Two isolates from chicken (#114 and #277; ToxoDB) and 3 from pigs (#114; ToxoDB) were the subjects for evaluation. The isolates were grown in cell culture under 2 different conditions: culture medium at pH 7.0 (neutral, without stress induction) or pH 8.0 (alkaline, stress inducing). After 4 days, the cultures were fixed and the events resulting from infection and induction were labeled. T. gondii cysts were labeled using Dolichos biflorus-FITC lectin (DBL-cysts) and free tachyzoites or vacuolar were labeled using an anti-T. gondii polyclonal antibody followed by an Alexa 594-conjugated secondary antibody (DBL-negative structures compatible with parasite structures - lysis plaques or vacuole). Differences in DBL-cysts formation in vitro in response to exogenous stress were observed between recombinant genotype isolates and the typical genotypes. The differences in conversion rates and the patterns of lysis plate production between genotype I/III isolates (#114) indicate that care should be taken when extrapolating the in vitro phenotypic characteristics of parasites from the same genotype., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Introduction: The present study aimed to assess the seroprevalence and spatial distribution of Neospora caninum and Toxoplasma gondii in dogs., Methods: Blood samples (n = 241) were collected and analyzed for the presence of anti-N. caninum and anti-T. gondii antibodies. The spatial distribution was evaluated using kernel density estimation (KDE)., Results: Anti-N. caninum and anti-T. gondii antibodies were detected in 24.06% (58/241) and 9.54% (23/241) of samples, respectively. A heterogeneous spatial distribution of positive dogs was observed across the city., Conclusions: These data are pivotal for better understanding the dynamics of infection caused by these protozoa in the canine population.

The purpose of this study was to perform genotypic characterization and to evaluate the virulence of Toxoplasma gondii obtained from aborted fetuses in an abortion outbreak in goats from northeastern Brazil. Brain samples from 32 fetuses were submitted to mouse bioassay for T. gondii isolation. Two isolates were obtained and subjected to genotypic characterization. Isolate virulence was evaluated using murine model in different doses (from 10 5 to 10 1 tachyzoites/mL). In genotyping, both isolates were classified as clonal lineage type II (genotype #1 ToxoDB) and showed to be virulent for mice. This is the first description of genotype #1 in cases of goat abortion, showing the circulation of virulent T. gondii isolate producing reproductive disorders in pregnant goat., (Copyright © 2018 Elsevier Inc. All rights reserved.)

The aim of this study was to determine the occurrence of infections due to Toxoplasma gondii and Neospora caninum and corresponding risk factors among dogs attended at veterinary clinics in the city of João Pessoa, Paraíba, northeastern Brazil. Blood samples were collected from 384 dogs that were attended at 34 veterinary clinics between April 2015 and May 2016. These two agents were diagnosed through the indirect immunofluorescence reaction (IFAT). Among the 384 animals evaluated, 37 (9.6%) were positive for T. gondii, with titers ranging from 16 to 512. Six dogs (1.6%) were positive for N. caninum, with titers of 50 to 200. Access to the streets (OR = 4.60; 95% CI = 1.74-12.20) and environments close to forested areas (OR = 2.79; 95% CI = 1.32-5.93) were found to be risk factors for T. gondii infection. The dogs attended at veterinary clinics in João Pessoa are exposed to infections caused by T. gondii and N. caninum. Dog owners should avoid having access to the street or contact with forest environments.

The aim of this study was to detect DNA and antibodies anti-Leishmania spp., Neospora caninum and Toxoplasma gondii in captive and free-range crab-eating fox (Cerdocyon thous) from northeastern Brazil. Twenty-five crab-eating foxes from different states of northeastern Brazil were sampled by this study. Blood samples were collected by cephalic or jugular vein punctures. The whole blood was submitted to PCR, and the sera samples to the serological analysis as follows: MAT for T. gondii, NAT for N. caninum, and ELISA for L. chagasi. The frequence of antibodies anti-T. gondii was 50% and 29.41% for free-range and captive wild canids, respectively. The frequence of antibodies anti-N. caninum observed by this study was 62.50% and 23.52% for free-range and captive wild canids, respectively. The frequence of antibodies anti-L. chagasi was 4.0% for captive wild canids. Co-infections cases were identified as follows: one captive wild canid seropositive for T. gondii and L. chagasi and two free-range animals seropositive for T. gondii and N. caninum. All PCR assays performed were negative for the pathogens analyzed. This study describes the presence of antibodies anti-T. gondii, N. caninum e L. chagasi in wild canids from northeastern Brazil and highlights the necessity of further studies on infectious diseases in free-range and captive wild canids.

The aim of this study was to evaluate the occurrence of Toxoplasma gondii in backyard pigs destined for human consumption in Pernambuco state, Brazil. Blood and tissue samples (liver, heart, brain, lung and diaphragm) were collected from 224 pigs from legal slaughterhouses and tested for T. gondii infection. Antibodies to T. gondii were found in the sera of 37.9% (85/224) by using the immunofluorescence antibody test (cut-off - 1:64). Tissue samples from seropositive pigs were bioassayed in mice. Tissue samples from seropositive pigs and from mice of the bioassay were submitted to histopathology, immunohistochemistry, polymerase chain reaction (PCR) and sequencing; 14.1% of pig tissue samples and 27.7% of bioassayed mouse samples were positive for T. gondii DNA, but all pig and mouse tissues were negative in histopathology analysis and immunochemistry. By using a risk assessment questionnaire, there was significant difference (p<0.001) in seroprevalence of 21.2% (reproducer) and 3.1% (finishing pig). These data serve as indicative of the sanitary conditions and risk of T. gondii infection for backyard pigs. Preventive measures must be implemented by health services to avoid toxoplasmosis human cases due to ingestion of pig meat.

Background: In Brazil, studies on animals and humans in mainland areas have shown that most strains of Toxoplasma gondii are pathogenic to mice and exhibit great genetic variability., Results: In this study, using a set of 11 PCR-RFLP and 15 microsatellite markers, we isolated and genetically characterised T. gondii strains from one cat and three rats on Fernando de Noronha Island. The cat had antibodies to T. gondii, which were revealed using a modified agglutination test (MAT, cut-off 1:25) and the seroprevalence among the 46 rodents was 15.2%. Viable T. gondii was isolated from one cat (TgCatBrFN1), two brown rats (TgRatnoBrFN1 and TgRatnoBrFN2) and one black rat (TgRatraBrFN1). Unlike the strains from mainland Brazil, these isolates were not pathogenic to outbred mice. The genotypes of these strains were compared with strains previously isolated on the island and in mainland Brazil. The analysis based on microsatellite data showed a limited genetic diversity of T. gondii on Fernando de Noronha Island with the majority of strains clustered into the following three groups: type II, III, and Caribbean 1., Conclusions: There was little variation among strains within the same group, suggesting that the majority of strains circulating on Fernando de Noronha are derived from only a few strains that were recently introduced to the island, likely from imported cats. Except for the strain belonging to the Caribbean 1 group that originates from northeast Brazil, there was little evidence that strains from the other groups were introduced to Fernando de Noronha via mainland Brazil.

Wild animals may play an important role in the transmission and maintenance of Toxoplasma gondii in the environment. The purpose of the present study was to isolate and genotype T. gondii from a free-ranging crab-eating fox (Cerdocyon thous-Linnaeus, 1766). A crab-eating fox in critical health condition was attended in a veterinary hospital in Recife, Pernambuco State, Brazil. The animal died despite emergency treatment. The brain was collected aseptically and destined for mouse bioassay. One isolate of T. gondii was obtained, and Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) was used to assess genetic variability at 11 markers (SAG1, SAG2, altSAG2, SAG3, BTUB, GRA6, c228, c292, L358, PK1 and APICO). A murine model was used to assess the virulence of the isolate. Using the PCR-RFLP, genotype ToxoDB #13 was identified, which is considered an atypical strain. The isolate was classified as avirulent in the murine model. This is the first study to report T. gondii infection in the crab-eating fox., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Little is known about toxoplasmosis in animals of the Fernando de Noronha Island, Brazil. Therefore, we investigated the prevalence of Toxoplasma gondii infection in the total population of pet cats (n=348), dogs (n=320), pigs (n=27), equines (n=101), as well as a significant portion of the population of feral cats (n=247) of the Island by Indirect Fluorescent Antibody Test. Anti-T. gondii IgG antibodies were found in 71.26%, 54.74%, 48.75%, 51.85% and 22.7%, of the pet and feral cats, dogs, pigs and equines, respectively, demonstrating a high prevalence of T. gondii infection in the wild and domestic animals of the Island. The Kernel intensity estimator showed a correlation between areas with high prevalence of infection in cats and occurrence of infection in the other studied species. We suggest that the island's health authorities should develop initiatives to reduce the population of cats and alert the island's population about the risk of T. gondii infection., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

This study aimed to investigate the frequency of anti-Toxoplasma gondii antibodies in serum from 629 chickens on 39 family farms in seven municipalities in the semiarid region, Pernambuco, Brazil, and to identify risk factors associated with T. gondii infection. The risk factors were studied in 421 samples from 29 farms. Anti-T. gondii antibodies were investigated by indirect fluorescent antibody test with a 1:16 cutoff. The frequency of positive chickens was 27.9% (176/629) and 94.8% of the farms studied had chickens infected by T. gondii. Multivariate analysis showed variables significantly associated with anti-T. gondii antibodies in serum: slaughter of animals on the farm, reproductive disorders in sheep, consumption of fetal adnexa and placentas by chickens, presence of sheep in the property and birth of sheep the property. The results suggest that there is a complex relationship between general management practices for different animal species raised on the same farm and the prevalence of T. gondii infection in chickens. In addition, the results draw attention to the risk of human infection by T. gondii via consumption of infected chicken meat, because the farming conditions and the low human development indices observed in the region studied result in inappropriate meat preparation practices.

Although the infection by Toxoplasma gondii in horses is usually asymptomatic, it may be characterized by the presence of tissue cysts and maintenance of detectable antibody titers. The aim of this study was to detect anti-T.gondii antibodies and to determine risk factors associated in sport horses from Paraíba State, Northeastern Brazil. A total of 138 apparently healthy sport horses of different age and gender were sampled. Anti-Toxoplasma gondii antibodies were detected in 16/138 (11.6%; 95% CI: 6.77-18.14%) horses by indirect immunofluorescence antibody assay (IFA). A significant association was found between the water source and seropositivity to T. gondii (p<0.05). The present study highlights the importance of providing good quality water to the animals.

Toxoplasmosis is a zoonotic disease of global distribution that affects all warm-blooded animals. The purpose of this investigation was to determine the prevalence of T. gondii infection and identify the risk factors associated with its occurrence in domestic ruminants raised on the island of Fernando de Noronha, Brazil, and to confirm that cattle and sheep raised in Fernando de Noronha Island present statistically different T. gondii prevalence rates. Serum samples were collected from sheep (n=240) and cattle (n=140) for the detection of antibodies by indirect immunofluorescence. Samples were collected from all the animals on all the farms. Risk factors were analyzed by univariate analysis and logistic regression. The prevalence rate of positive sheep was 85.0% while that of cattle was 10.7%. A multivariate analysis revealed that the site of contact of sheep with felines was a risk factor. For cattle, the risk factors identified in this study were: extensive farming system, water source, more than three cats per farm, and the presence of rats in feed storage locations. The findings revealed a significant difference in the prevalence rates in sheep and cattle raised in this insular environment.

The present study aimed to conduct a serosurvey of toxoplasmosis in domestic rabbits of Northeastern Brazil. Blood samples and tissue fragments (brain, heart and diaphragm) were collected from 150 and 54 rabbits from the state of Pernambuco, Brazil, respectively. The serum samples were subjected to serological analysis (Modified Agglutination Test - MAT) and the tissue samples were assessed by PCR and histopathological analysis. Data collected through questionnaires were subjected to analysis of risk factors. According to the MAT and the PCR results, 6.7% (10/150; CI 3.2%-11.9%) of the rabbits were positive for anti-T. gondii antibodies and 9.25% (5/54) of the tissue fragments were positive for T. gondii DNA, respectively. Lesions associated with T. gondii infection, mainly characterized by granuloma, mononuclear cell infiltrates, degeneration areas and necrosis in brain and heart, were detected in the histopathological analysis. The risk factors associated with T. gondii infection identified in the present study were homemade food (odds ratio = 39.00) and contact between cats and rabbits (odds ratio = 52.00). This is the first report of toxoplasmosis in rabbits of Northeastern Brazil. The management problems identified in the present study must be corrected to reduce the frequency of positive animals in herds of rabbits.

Toxoplasma gondii isolates from Brazil have a different phenotypic and genotypic pattern, with predominance of virulent isolates and recombinant genotypes, compared to the North Hemisphere. Considering that a new T. gondii genotype, non-pathogenic to mice, was previously identified from free-range chickens from the Fernando de Noronha Island, Brazil, this study aimed to identify genotypes of this parasite in tissue samples of feral cats (Felis catus) from this Brazilian Island. Anti-T. gondii IgG antibodies were detected in 18/31 (58%) feral cats. Two non-virulent T. gondii isolates were obtained by mouse bioassay. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico) and an atypical strain of T. gondii (ToxoDB #146) was identified. This is the first report of this genotype in feral cats., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of the present study was to investigate the prevalence of antibodies against Toxoplasma gondii in turkeys and chickens on family farms in the semi-arid region of the state of Pernambuco, Brazil. In order to do so, 204 sera samples from turkeys were analyzed using the Modified Agglutination Test (MAT) and 322 sera samples from chickens were assessed using the Immunofluorescence Assay (IFA). Twenty-eight properties contained turkeys and 23 of these properties also contained chickens. The relative frequency of positive turkeys was 11% (21/204), with 46.6% (13/28) of the properties containing at least one positive turkey. The frequency of positive chickens was 25.8% (83/322), with 95.6% (22/23) of the properties containing at least one positive chicken. Based on the results of the present study, it was possible to conclude that turkeys can serve as an indicator of environmental contamination by oocysts of T. gondii. However, they are less effective than chickens bred in the same conditions. The increasing demand from consumers for naturally produced products should worry local sanitary authorities due to the high prevalence of antibodies against T. gondii found in this type of rearing system, particularly among chickens.

The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chickens bred in the metropolitan area of Recife, Brazil. In total, 212 serum samples were collected from 16 properties, and 12 backyard chickens were collected in the six sanitary districts of Recife. An indirect immunofluorescence assay (IFA) was used to investigate the occurrence of anti-Toxoplasma gondii antibodies. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in brain, heart, liver and lung specimens. Of the samples analyzed by serology, 86/212 (40.56%) were positive; of the samples analyzed by PCR, 2/12 (16.7%) were positive, with both samples positive by both tests (serological and molecular). The presence of antibody anti-T. gondii and parasite DNA in tissues of these animals are worrying aspects for public health because there is a risk of transmission of the parasite to humans through eating undercooked or raw meat. Based on the results, the adoption of preventive measures to prevent the cats access to the chickens creations should be encouraged, since these animals were identified in most of the studied properties.

The aim of the present study was to investigate the occurrence of Toxoplasma gondii among horses and its associated risk factors in Alagoas, Brazil. In total, 440 samples from 36 properties in 23 districts of the state of Alagoas were studied, covering the Leste, Agreste and Sertão mesoregions. Risk factors were evaluated through the application of an investigative questionnaire that focused on the productive, reproductive and sanitary management of herds. T. gondii infection were assayed using the immunofluorescence assay (IFA) with a cutoff point of 64; 14.4% (95% CI: 11.0%-17.8%) of - horses were seropositive. A significant association was determined between anti-T. gondii antibody presence and the consumption and storage of hay (OR = 2.08 / 95% CI: 1.20-3.62). This is the first report of T. gondii infection among horses in the state of Alagoas, Brazil.

The aim of this study was to investigate occurrence of Toxoplasma gondii in sheep slaughtered in the state of Alagoas, Brazil, by means of different diagnosis techniques. Serum samples and tissues from 100 slaughtered sheep were used. To detect antibodies, the indirect immunofluorescence antibody test (IFAT) was used, and tissues from seropositive animals (cut-off ≥1:64) were submitted to Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC). To assess the concordance between the direct techniques, the kappa test was used. In the IFAT, it was observed that 14% (14/100) of the ovine samples were serum-positive. In the PCR, 21.43% (3/14) of the animals were positive and in IHC, it was observed that 7.14% (1/14) were positively stained for T. gondii in cerebral tissue. Histopathologically, the predominant finding was the presence of mononuclear cell infiltrate in the heart and a perivascular cuff in the cerebrum and cerebellum. The concordance between the direct diagnosis techniques was moderate (k=0.44). Thus, it is important to use different direct techniques in diagnosing toxoplasmosis in naturally infected sheep.

The aim of the present study was to isolate and genotype Toxoplasma gondii from pigs slaughtered for human consumption in northeastern Brazil. Indirect immunofluorescence antibody test (IFAT) was used to screen positive pigs. Tissues samples of animals with antibody titers ≥64 were submitted to bioassay in mice. One isolate of T. gondii was obtained, and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, using 11 markers (SAG1, SAG2, altSAG2, SAG3, BTUB, GRA6, c228, c292, L358, PK1, and APICO), was applied to evaluate the genetic variability. DNA from reference strains was used as a positive control. By means of genetic analysis, genotype ToxoDB #65 was identified, which is considered an atypical strain. This is the first record of genotype #65 in pigs. Thus, further studies in this region are necessary to determine the genetic variability of T. gondii in pigs and possible impact on public health.

The aim of the present study was to detect the genomic DNA of Toxoplasma gondii in milk samples from naturally infected goats in the state of Pernambuco, (Brazil). In total, 248 blood serum samples were collected and processed from lactating goats and then submitted to a search for antibodies to T. gondii through the indirect immunofluorescence reaction. Samples with a score of 64 or more were considered positive. In total, 248 milk samples were collected and processed from the same group of goats in order to study the DNA of T. gondii using the polymerase chain reaction (PCR) technique. In the serum samples, 56/248 (22.58%) of the animals were positive, whereas the DNA of the parasite was detected in 15/248 (6.05%) of the milk samples. Five of these 15 samples were animals who were also positive in the serology. This study reports the first occurrence of the elimination of T. gondii from the milk of naturally infected goats in the north-east of Brazil. It is suggested that the consumption of in natura goat milk may constitute a potential risk to the health of milk consumers in this region., (© 2013 Blackwell Verlag GmbH.)

The aims of this survey were to determine the flock-level and animal-level prevalences of anti-Toxoplasma gondii antibodies in sheep in the State of Paraíba, Northeastern Brazil, and to identify risk factors. Blood samples were collected from 540 sheep in 63 flocks in 14 municipalities in the Sertão mesoregion. To serologically diagnose T. gondiii nfection, the indirect immunofluorescent antibody test (IFAT) was used. Flocks with at least one seropositive animal were considered positive. Among the 63 flocks, 28 (44.44%) were positive, and 60 (11.11%) of the 540 animals were seropositive. Variable flock size > 25 animals was identified as a risk factor (odds ratio = 3.2; 95% CI = 1.09 - 9.34; P = 0.033). The results from this survey demonstrate that T. gondii is spread among sheep in the State of Paraíba.

The hoary fox (Pseudalopex vetulus) is a wild canid native to Brazil and is commonly found in the semiarid northeastern area living in contact with cattle. The main purpose of this study was to investigate the presence of Neospora caninum and Toxoplasma gondii DNA in hoary foxes, in the state of Paraíba, Brazil. Brain tissue samples were collected from 49 hoary foxes. From the samples, DNA extraction and the polymerase chain reaction (PCR) were performed using specific primers for N. caninum and T. gondii. The prevalences found were 14.3% (7/49) for T. gondii and 12.2% (6/49) for N. caninum. The molecular identities of the amplified products were confirmed by means of the sequencing reaction. This study demonstrated the presence of N. caninum and T. gondii DNA in free-ranging hoary foxes in Brazil for the first time, thus confirming that this species is an intermediate host., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The aim of this study was to identify risk factors associated with prevalence of Toxoplasma gondii infection in captive capuchin monkeys at a facility in the northeastern Brazil. Serum samples from 116 bearded capuchin (Sapajus libidinosus), nine blonde capuchin (Sapajus flavius), five black-capped capuchin (Sapajus apella), and four capuchin monkeys (Sapajus spp.) were tested for T. gondii antibodies using the modified agglutination test (MAT, cut-off ≥25); antibodies were found in 85.3% (99/116) of S. libidinosus, 55.6% (5/9) of S. flavius, 80.0% (4/5) of S. apella, and 75.0% (3/4) of S. spp. The risk factors associated with T. gondii seropositivity were ingestion of raw meat [OR = 4.13 (1.26; 13.50)] and old age [OR = 4.90 (1.70; 14.13)]. Results indicate a very high T. gondii seropositivity in these primate populations. To minimize exposure to T. gondii raw meat should not be fed to these animals., (© 2015 Wiley Periodicals, Inc.)

The aim of the present study was to detect Toxoplasma gondii DNA in raw milk samples of goats and sheep of local breeds from the semi-arid region of the states of Pernambuco and Paraíba, Northeastern Brazil. Serum and milk samples were collected from 243 animals (186 goats and 57 sheep). The Indirect fluorescent antibody test (IFAT) was used to search for anti-T. gondii IgG antibodies with a cutoff of 64. Subsequently, the raw milk samples were subjected to DNA extraction and PCR to detect DNA of T. gondii. The IFAT results showed a 6.58% (16/243) positivity when all the samples were considered and a positivity of 15.78% (9/57) and 3.76% (7/186) for goats and sheep samples, respectively. The PCR assay detected T. gondii DNA in 2.06% (5/243) of all the samples tested. All the PCR positive samples were from goats. This result shows the importance of adopting measures of flock's sanitary management and avoiding the consumption of raw milk may constitute a potential risk to the health of milk consumers in this region., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The aim of this study was to estimate the prevalence of genomic DNA of Toxoplasma gondii in semen samples from commercial rams in artificial insemination centres in Brazil, as well as in fresh semen from rams in the northeast of Brazil. In total, 108 semen samples were obtained from artificial insemination centres, and genomic DNA of T. gondii was detected in 24 of 108 (22.2%). The prevalence of antibodies anti-Toxoplasma gondii among sheep on rural properties was 9.2% (10/109), and 100% of the semen samples of these animals were positive in the PCR for T. gondii DNA. The molecular identity was confirmed through sequencing, which indicated 99.9% similarity with the T. gondii DNA sequences stored in the GenBank. This study reports the first occurrence of T. gondii DNA in the semen of rams, which came from artificial insemination centres in Brazil, as well as the occurrence of T. gondii DNA in the fresh semen of naturally infected rams in the northeast of Brazil., (© 2014 Blackwell Verlag GmbH.)

Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii that affects homeothermic animals, including humans. Felines are considered the definitive host of this parasite, while other animals act as intermediate hosts. The purpose of this study was to assess the frequency of anti-T. gondii IgG antibodies in bovines in the state of Pernambuco, Brazil. Serum samples (n = 427) from animals in 13 municipalities of the coastal forest/plantation region of the state were analyzed using the immunofluorescent antibody test (IFAT). The overall results revealed a prevalence rate of 16.63% (27/427). High percentages of positivity were found among animals aged 25 to 36 months (28.57%; 30/42) and in males (22.22%; 2/9). The present findings suggest that bovine toxoplasmosis is endemic in the area under study.

Felids are important in the epidemiology of Toxoplasma gondii because they are the only hosts that can excrete the environmentally resistant oocysts in their feces. Cats acquire T. gondii infection in nature by ingesting tissues of small mammals and birds. Serum samples of 223 feral marsupials and 174 feral rodents captured in 7 segments of the Atlantic Forest of the State of Pernambuco, northeastern region of Brazil, and in urban areas of the municipality of Recife were examined for antibodies to T. gondii by the modified agglutination test (MAT). Antibodies (MAT ≥ 25) were found in 6.7% (15 of 223) of the marsupials and 5.7% (10 of 174) of the rodents. No association was observed between seropositivity in marsupials or rodents and sex, age, or different areas of collection (P > 0.05). This is the first study on the seroprevalence of T. gondii in marsupials and rodents performed in the Atlantic Forest of the northeastern region of Brazil. The presence of antibodies to T. gondii are reported for the first time in long-furred woolly mouse opossum ( Micoureus demerarae ), murine mouse opossum ( Marmosa murina ), brown four-eyed opossum ( Metachirus nudicaudatus ), and gray short-tailed opossum ( Monodelphis domestica ).

The objective was to characterize the transmission of Toxoplasma gondii in goats experimentally infected vaginally with semen contaminated with the CPG strain (genotype III). Ten female goats were randomly allocated into 2 groups (G1 and G2), each with 5 animals, and inseminated during estrus. Goats in G1 were inseminated with semen containing 1 × 10(5) tachyzoites, whereas those in G2 (control) were inseminated with semen free from tachyzoites (insemination = day 0). In G1, seroconversion (indirect immunofluorescence reaction) and DNA (polymerase chain reaction) in the blood was present in 4/5 and 3/5, respectively, from the 7th day. In G2, all goats were negative in all tests. Embryonic reabsorption occurred in 4 of 5 goats from G1 between days 21 and 49. In conclusion, artificial vaginal insemination with semen containing tachyzoites of T. gondii -infected goats and is a potential transmission route of this parasite through semen.

The aim of the present study was to investigate antibodies to Toxoplasma gondii in the serum of mules and donkeys bred in the northeast of Brazil. In total, 483 samples were used (395 mules and 88 donkeys) from 4 states (Pernambuco, Rio Grande do Norte, Paraíba, and Sergipe). The indirect immunofluorescence reaction (IFI) technique was used to investigate antibodies to T. gondii with a cut-off point of 64. Positive frequencies of 23.8% and 43.2% were recorded for mules and donkeys, respectively. The state of Pernambuco had the highest prevalence of positive samples (29%) with statistically significant differences for species (P < 0.001) and state (P = 0.048). This is the first study of antibodies to T. gondii in mules and donkeys in these 4 states of the northeastern region of Brazil and serves as a warning to health authorities regarding the risks of ingesting equine meat.

Fernando de Noronha is an archipelago of 21 islands and islets in the Atlantic Ocean, state of Pernambuco, Brazil, which has a varied biodiversity including alien species or sinantropic animals. The objective here was to determine the seroprevalence of Toxoplasma gondii in domestic and wild animals from Fernando de Noronha archipelago, Brazil. Between July 2007 and May 2010, blood samples were collected from 764 animals (533 domestic and 231 wild animals). Sera were tested by the indirect fluorescence antibody test (IFAT) or the modified agglutination test (MAT), or by both. Antibodies to T. gondii were found in 80 (80.0%) of 100 chickens ( Gallus domesticus ), 3 (3.0%) of 100 cattle ( Bos taurus ), 59 (60.8%) of 97 sheep ( Ovis aries ), 9 (81.8%) of 11 goats ( Capra hircus ), 7 (43.7%) of 16 horses ( Equus caballus ), 70 (59.3%) of 118 cats ( Felis catus ), 36 (39.6%) of 91 dogs ( Canis familiaris ), 13 (38.2%) of 34 black rats ( Rattus rattus ), and 157 (79.7%) of 197 cattle egrets ( Bubulcus ibis ). Results indicate endemic infection by this zoonotic parasite among the animal and avian fauna in this archipelago from Brazil.

The object of this study was to investigate Toxoplasma gondii antibodies and parasite DNA in pigs in the state of Pernambuco, Brazil. Blood samples were collected from 305 slaughtered pigs in 11 municipalities, and their sera were tested for T. gondii antibodies using the indirect fluorescent antibody test (IFAT, cutoff 1:64); 38 (12.5%) samples were positive. Attempts were made to detect T. gondii DNA in the heart tissue of seropositive pigs using the B 1 gene and PCR; 21 (55.2%) of the 38 hearts were positive. This is the first detection of T. gondii DNA in tissues of serologically positive swine in the State of Pernambuco, Brazil.

The aim of this research was to study the contribution of Toxoplasma gondii to reproductive failure using nested PCR and histopathological examination of fetuses, stillborns and placentas. We examined 245 organs of fetuses and 28 placentas from 35 abortions and stillborns from naturally occurring miscarriages in sheep in the State of Pernambuco, Brazil. At necropsy, fragments of brain, cerebellum, medulla, lung, heart, spleen, liver and placenta were taken for nested PCR and histopathological tests. Pathological examination revealed macroscopic lesions, suggesting T. gondii infection in 5/35 (14.3%) of the placentas. The histopathological examination revealed no lesions characteristic of toxoplasmosis in the organs investigated. In the five placentas, lesions consistent with toxoplasmosis were observed as an inflammatory non-suppurative infiltrate, along with multiple necrosis and mineralization. Nested PCR showed three aborted fetuses and two stillborns (14.3%) to test positive for T. gondii, with DNA amplification in all organs and the placenta, especially the heart and the placenta, which are the tissues of choice. This study substantiates the theory that T. gondii is involved in miscarriages and stillbirths and in the placentas of naturally infected sheep in Brazil. Such findings have not previously been described in the national literature., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Introduction: Goats are considered very susceptible to infection by Toxoplasma gondii and when this occurs during pregnancy, it may cause fetal death with subsequent fetal resorption, abortion, mummification, stillborn or the birth of weak goats. The objective of this work was to determine the occurrence of and identify risk factors for T. gondii infection in goats in different mesoregions in the State of Alagoas., Methods: The research was conducted on 24 goat breeding farms in 10 municipalities. A total of 454 blood samples were examined for anti-T. gondii antibodies by indirect immunofluorescence antibody test. To evaluate the risk factors of toxoplasmosis in goats, questionnaires were applied analyzing the farm's production system and nutritional, reproductive and sanitary management., Results: Disease occurrence was 39% with 95.8% of farms presenting seropositive animals. Significant associations were observed for mesoregion (OR = 0.23; 95%CI = 0.09 - 0.57), age (OR = 0.36; 95%CI = 0.20 - 0.64), semi-intensive herd management (OR = 8.70; 95%CI = 1.87 - 40.43), access of cats to water provided for goats (OR = 3.38; 95%CI = 1.89-6.02) and cats feeding on placental remnants (OR = 2.73; 95%CI = 1.38 - 5.40)., Conclusions: Toxoplasma gondii infection is disseminated in the State of Alagoas and the adoption of a program that monitors infection foci is required to reduce the risk factors identified in the present study.

The aim of this study was to characterize the reproductive disorders in the acute and chronic phases in ewes experimentally infected with different doses of Toxoplasma gondii during artificial insemination occurred. Animals (n=41) were divided into three experimental groups: in the group 1 (G1, n=15), animals were inseminated using contaminated semen containing 6.5×10⁴ tachyzoites; in the group 2 (G2, n=15), animals were inseminated with contaminated semen containing 4×10⁷ tachyzoites and in the group 3 (G3, n=11), animals were inseminated using tachyzoite-free semen, serving as control group. Parasitemia and seroconversion were observed in 28 of 30 and 20 of 30, respectively, from the seventh day after infection. Embryonic reabsorption was observed in the acute phase in ewes from G1 and G2. Persistent anestrus, hydrometra, mucometra and follicular cysts were observed in the second phase of the experiment in animals from G1 and G2. Histopathological lesions similar to those of toxoplasmosis were found in the placentas. In conclusion, artificial insemination using semen containing experimentally added tachyzoites can establish toxoplasmosis in ewes and cause reproductive pathologies during the acute and chronic phases of the disease., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Worldwide comparison of Toxoplasma gondii isolates from free-range chickens ( Gallus domesticus ) has indicated that T. gondii isolates from Brazil are phenotypically and genetically different from isolates from other countries; most strains from Brazil are pathogenic to mice, there is great genetic variability, most isolates are nonclonal, and Type II is absent or rare. The prevalence of T. gondii in 50 free-range chickens from the island of Fernando de Noronha, Brazil (this island is 350 km from the mainland) was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT); 42 (84%) chickens had titers of 1ratio5 in 2, 1ratio10 in 4, 1ratio20 in 3, 1ratio40 in 6, 1ratio80 in 6, 1ratio160 in 5, 1ratio320 in 3, and 1ratio640 or higher in 13 chickens. Hearts of 40 seropositive chickens were bioassayed individually in mice. Toxoplasma gondii was isolated from 24 chickens with MAT titers of 1ratio5 or higher; the isolates were designated TgCKBr210-233. None of the isolates was pathogenic for mice. The restricted fragment length polymorphism using 10 markers revealed 6 genotypes, including the Type II, Type III, and 4 new chicken genotypes (#59-#62) that were different from genotypes so far reported in Brazil. All 24 isolates were successfully genotyped; 15 isolates were Brazil chicken type #59, 1 type #60, 1 type #61, 1 type #62; 5 were Type II (with Type I allele at the Apico locus); and 1 isolate was clonal Type III. Results in this study indicate that T. gondii on this island consists of unique genotypes as well as clonal genotypes that are dominant in Europe and North America.

Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii that affects reproductive performance in small ruminants. Although the T. gondii life cycle is well understood since 1960s, several aspects related to its infection remain unclear. In the present study we hypothesized that sheep inseminated with T. gondii-contaminated semen would develop toxoplasmosis. In order to test that hypothesis, 41 sheep were experimentally infected with semen spiked with the organism. Females were divided in three groups (G1-G3): (a) females in G1 group were inseminated with semen containing 6.5 x 10(4) tachyzoites; (b) females in G2 group with semen containing 4 x 10(7) tachyzoites; and (c) females in G3 group with tachyzoite-free semen (control group). To confirm T. gondii infection via semen, serological tests were performed using indirect immunofluorescence reaction and the detection of parasite DNA in the blood stream using the nested PCR test. While in G1 group only 5/15 (33.3%) of the females presented seroconversion, all sheep in G2 15/15 (100%) seroconverted. The nested PCR test showed that 14/15 (93.3%) of the females in the G1 and 14/15 (93.3%) in the G2 group were positive for T. gondii while in the G3 group all samples were negative. In addition, ultra-sound test evidenced that in sheep presented embryonic reabsorption in animals from the infected groups. In conclusion, insemination using fresh semen experimentally contaminated with different infectant doses of T. gondii tachyzoites was able to infect sheep, leading to the possibility of toxoplasmosis transmission via semen., ((c) 2010 Elsevier B.V. All rights reserved.)