Prevalence of Neospora caninum and Toxoplasma gondii in sheep and dogs from Guarapuava farms, Paraná State, Brazil

Sheep and dog blood samples were collected from nine farms in the county of Guarapuava, Paraná, Brazil. The indirect fluorescent antibody test (IFAT) was used to detect Neospora caninum and Toxoplasma gondii antibodies. Herein, serum samples from 305 sheep were evaluated, being 29 (9.5%) and 157 (51.5%) seropositives to N. caninum and T. gondii, respectively. Seven (29.1%) and five (20.8%) out of 24 dogs were seropositives to N. caninum and T. gondii, respectively. There were no differences among the sheep serology for N. caninum and reproductive problems, management and animal feeding variables, neurological problems and presence of other animals species on the farm (P>or=0.05). The simultaneous frequency of antibodies between N. caninum and T. gondii was 5.2% in the herds. Age, breed, farm size, semi-intensive activity, mineral salt supplementation, water origin, stage of the pregnancy when reproduction problems occurred, neurological problems in lambs, presence of rodents in the food room and pasture cat access were identified as associated factors for the occurrence of toxoplasmosis in sheep (P<0.05). There were no differences among the seropositivity in dogs for N. caninum and T. gondii and breed, age and sex (P>or=0.05). The present work is the first report on serum prevalence of N. caninum in sheep from the state of Paraná, Brazil.     

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

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

Serological profile of Toxoplasma gondii and Neospora caninum infection in commercial sheep from São Paulo State, Brazil

Neosporosis and toxoplasmosis are two important infections in young and adult sheep, leading to low production and abortion. This study aimed to determine the frequency of antibodies to Toxoplasma gondii and Neospora caninum in sheep from the eastern region of S?o Paulo State, Brazil. Serum samples (382) were collected from the sheep and assayed for T. gondii through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT), and for N. caninum antibodies, through IFAT, with cut-off titers equal to 16 (T. gondii) and 25 (N. caninum). All frozen samples were sent to the Center for Zoonoses Research (NUPEZO), Department of Veterinary Hygiene and Public Health (DHSVP), FMVZ, UNESP, for serological tests. A total of 71/382 (18.6%) samples reacted to T. gondii, especially at titers 16 (28; 39.4%), 64 (15; 21.1%), 256 (21; 29.6%) and 1024 (6; 8.5%) by MAT, and 16 (34; 47.9%), 64 (18; 25.4%), 256 (14; 19.7%) and 1024 (5; 7%) by IFAT. As regards N. caninum, 49/382 (12.8%) samples reacted at titers 25 (17; 34.7%), 50 (11; 22.5%), 100 (11; 22.5%), and ≥ 200 (10; 20.4%). These animals presented infection but no clinical signs. Six and ten animals had high titers for toxoplasmosis and neosporosis. No significant association was observed between antibodies for both parasites (P=0.535) according to Fisher's exact test, and no correlation was found between T. gondii (MAT) and N. caninum antibody titers (r=-0.0068; P=0.895), T. gondii (IFAT) and N. caninum antibody titers (r=-0.0025; P=0.961). Thus, T. gondii and N. caninum infections were observed in farms located in S?o Paulo State, where sheep play an important economical role for the national and regional business.     

Seroprevalence of Toxoplasma gondii and Neospora caninum in sheep intended for human consumption in the Rondônia state,Western Brazilian Amazon

The aim of this study was to carry out a seroepidemiological study of anti-T. gondii and anti-N. caninum antibodies in serum samples from sheep intended for human consumption in the Rondônia state, in the Western Brazilian Amazon, and identify possible risk factors associated with seropositivity. The analysis of the 616 sheep serum samples revealed animal-level seroprevalence of T. gondii in the order of 52.4 % and of N. caninum of 60.6 %. Prevalence of co-infections was 33.4 %. The herd-level seroprevalence was 100 % for both the parasites. Applying of multivariate logistic regression analysis, test of the association between seroprevalence of T. gondii infection and the potential predictors showed that feed, fetal malformation, abortion, neurological problems (lambs), presence of rodents were the significant (P < 0.05) predictors. The variables presence of rodents, cats, wild animals and main activity showed statistical association (P < 0.05) with the seroprevalence of N. caninum. Due to the high prevalence of T. gondii and N. caninum found in this study, the absence of specific slaughterhouses for sheep and raw or undercooked sheep meat for human consumption we concluded that sheep can represent an important source of infection for humans.     

Evaluation of Toxoplasma gondii and Neospora caninum infections in sheep from Uberlândia, Minas Gerais State, Brazil, by different serological methods

Toxoplasmosis and neosporosis have been recognized as economically important diseases with considerable impact on the livestock industry. Considering the scarce information on the occurrence of Toxoplasma gondii and Neospora caninum infections in sheep from Uberlandia, Minas Gerais State, Brazil, this study aimed to investigate the frequency of antibodies against these parasites in sheep sera from this region by using different serological methods. A total of 155 sheep serum samples were analyzed by the indirect fluorescence antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) for the detection of IgG against T. gondii and N. caninum. Seroreactivity by IFAT showed 80% of samples with titers between 512 and 2048 for T. gondii (cutoff ≥ 64) and 78% presenting titers between 50 and 200 for N. caninum (cutoff ≥ 50). Seroreactivity by ELISA showed 75% of samples with ELISA index (EI) between 2.0 and 3.0 for T. gondii (cutoff ≥ 1.3) and 54% presenting EI between 1.3 and 2.0 for N. caninum (cut off ≥ 1.3). Discordant results by both tests were analyzed by immunoblot, resulting in a total seropositivity of 61% for T. gondii and 23% for N. caninum, with 41% to T. gondii only, 3% to N. caninum only, and 20% to both parasites. There was a significant positive association between seropositivity to T. gondii and age over one year (P<0.001), but such association was not found for N. caninum infection. In conclusion, as T. gondii and N. caninum infections are simultaneously present in sheep flocks of this region, it should be emphasized the importance to carry out a regular monitoring of Toxoplasma infection due to its high prevalence, its zoonotic potential and induction of reproductive disorders leading to economic losses. For neosporosis, sheep farmers should be instructed about the presence of the parasite in the flock, its risk factors and potential abortifacient role in sheep. Differential flock management could be valuable tool to establish the association of serological positivity and reproductive disease induced by N. caninum in sheep.     

Vertical and horizontal transmission of Neospora caninum in dairy herds in Québec

Neospora caninum is an important cause of abortion in dairy cattle. The objective of this observational study was to estimate the rate of vertical transmission of N. caninum in dairy herds in Québec and to investigate horizontal transmission in the same herds. The genealogy of cows from 23 dairy herds were examined. Prevalence of seropositive animals in herds studied varied from 4.3% to 61.8% (average, 21.9%). The overall rate of vertical transmission was estimated to be 44.4%, varying from 0% to 85.7%. Seven cases of horizontal transmission were identified in 6 of the 23 herds studied. Estimated vertical transmission rate varied from herd to herd, but appeared to be higher in herds with a high prevalence of seropositive animals. Although horizontal transmission was identified in 6 herds, it does not appear to be the major route of infection for N. caninum.     

Seroprevalence of Toxoplasma gondii infection in sows in Münsterland, Germany

A cross-sectional survey was performed to obtain current data on the seroprevalence of Toxoplasma gondii infection in breeding sows in Münsterland, Germany. Antibodies to T. gondii were detected by ELISA in 140 (9.3%) of the 1500 sows examined. Forty-two (56%) of the 75 surveyed farms harboured at least one seropositive animal. In 3 (5%) of 62 farms where at least 10 sows had been examined > or = 30% of the animals were seropositive.     

Do free-ranging common brushtail possums (Trichosurus vulpecula) play a role in the transmission of Toxoplasma gondii within a zoo environment?

To investigate the possible role of common brushtail possums (Trichosurus vulpecula) in the transmission of Toxoplasma gondii within a zoo environment, a serological survey of a free-ranging population resident within Taronga Zoo, Sydney, Australia was undertaken using the modified agglutination test (MAT). For comparison, the seroprevalence of T. gondii antibodies was also assessed in a possum population inhabiting a felid-free, non-urban woodland habitat. Six of 126 possums (4.8%) from the zoo population had antibodies to T. gondii with a MAT titre of 25 or higher, while in contrast, all of the 17 possums from woodland were seronegative. These observations suggest that possums were at a higher risk of exposure to the parasite as a consequence of co-existing with domestic, stray and captive felids associated with urbanisation. Screening of captive felids at the zoo indicated 16 of 23 individuals (67%) and all 6 species were seropositive for T. gondii, implicating them as a possible source of the parasite within the zoo setting. In addition captive, non-felid carnivores including the chimpanzee (Pan troglodytes), saltwater crocodile (Crocodylus porosus), dingo (Canis lupis) and leopard seal (Hydrurga leptonyx) were tested for the presence of T. gondii antibodies as these species predate and are a leading cause of death amongst zoo possums. In total, 5 of 23 individuals (22%) were seropositive, representing 2 of the 4 carnivorous species; the dingo and chimpanzee. These data suggest that carnivory was not a highly efficient pathway for the transmission of T. gondii and the free-ranging possum population posed minimal threat to the health of zoo animals.     

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

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

What Do We Know About Risk Factors for Infection in Humans with Toxoplasma gondii and How Can We Prevent Infections?

Fourteen case–control studies evaluating the importance of different risk factors for infection of humans with Toxoplasma gondii were identified. Surprisingly, up to two‐thirds of infections could not be explained by risk factors such as consuming raw or undercooked meat or poor kitchen hygiene. Few studies reported population attributable risk, and when reported, it was low. Protocols including questionnaires used for collecting data, age and socio‐economic groups studied, and sample sizes varied among studies. There is a need for carefully planned, case–control studies to identify sources of infection with T. gondii in humans. New risk factors, such as the manufacture of organic meat, and possible ways to sterilize meat to inactivate T. gondii tissue cysts are discussed.     

Seroprevalence of Neospora caninum antibodies and its consequences for reproductive parameters in dairy cows from Dakar–Senegal,West Africa

The aim of this study was to determine the seroprevalence of Neospora caninum antibodies and its effects on reproductive parameters in cows in intensive dairy herds in Dakar. Randomised blood samples were taken for serology from 196 cows in four herds with a history of sporadic abortion. All of the sera were assayed for antibodies against N. caninum, Candida guillermondii, Coxiella burnetii, and Chlamydophila sp. The associations between serostatus and reproductive parameters (abortion, number of inseminations to conception, and calving to conception interval) were assessed over a period of 5 years (2004–2008). The seroprevalence of N. caninum antibodies in dairy cattle was 17.9%. The local Gobra breed and crossbreeds had higher levels of N. caninum antibodies than exotic breeds (p < 0.05). For the most recent pregnancies, seropositive cows required more inseminations to establish conception than seronegative cows (p < 0.05). The results indicate that dairy cattle from Dakar are exposed to N. caninum. Neosporosis should, therefore, be systematically considered as a cause when the calving to conception interval is prolonged.     

Toxoplasma gondii infection in cats from São Paulo state, Brazil: seroprevalence, oocyst shedding, isolation in mice, and biologic and molecular characterization

Cats are the most important hosts in the epidemiology of Toxoplasma gondii infections in humans and animals. Serologic and parasitological prevalence of T. gondii were determined in 237 cats from 15 counties in S?o Paulo state, Brazil. Antibodies to T. gondii were found in a 1:25 dilution of serum of 84 (35.4%) out of 237 cats by the modified agglutination test (MAT). Samples of brain, heart, tongue, and limb muscles (total 50 g) of 71 of the seropositive cats were pooled for each cat, digested in pepsin and bioassayed in mice. Faeces (1 g) from the rectum of each cat were examined microscopically for T. gondii-like oocysts and verified by bioassay in mice; T. gondii oocysts were found in the faeces of three (1.3%) of 237 cats. T. gondii was isolated from tissue homogenates of 47 cats. The DNA obtained from these 47 tissue isolates was characterized using the SAG2 locus: 34 (72.4%) isolates were type I, 12 (25.5%) were type III and one (2.1%) was mixed with types I and III. No type II isolates were detected. Most (23/34) of the type I isolates killed all infected mice and 7 of 12 type III isolates did not kill infected mice. Characterization of the SAG2 locus directly from tissue homogenates from 37 of 46 cats was successful. Genotypes obtained from these primary samples were the same as those from the corresponding isolates obtained in mice. Genotyping of the three oocyst isolates revealed that two were type I and one was type III. Molecular and biologic characteristics of T. gondii isolates from animals from Brazil are different from those from other parts of the world.     

Environmental pollutants and diseases of sexual development in humans and wildlife in South Africa: harbingers of impact on overall health?

This study deals with disorders of sexual development in humans, wildlife and animals in an urban nature reserve (RNR) and a currently DDT-sprayed malarial area. High levels of oestrogenic chemical residues in water, sediment and tissue; skewed sex ratios; reduced biodiversity; gonadal malformations in sharptooth catfish and freshwater snails; intersex in catfish; and impaired spermatogenesis in catfish and striped mouse are of serious concern in the RNR. Persistent eggshell thinning in African darter eggs, intersex in male Mozambican tilapia, follicular atresia in females and impaired spermatogenesis in males following laboratory exposure of parent fish to environmentally relevant DDT and DDE concentrations, and abnormalities in freshwater snails were found in the DDT-sprayed area. Human studies related to DDT exposure indicated impaired semen quality, a weak association with sperm chromatin defects and higher risks for external urogenital birth defects in those who were born to mothers whose houses were sprayed and those who were homemakers (stay at home mother) instead of being employed. These findings indicate that diseases of sexual development occurred in both human and wildlife populations exposed to environmental endocrine disruptor chemicals in South Africa. The chemical mixtures, possibly related to disorders of sexual differentiation (DSD), were very different between the two. However, DSD occurred concurrently in the malarial area, possibly indicating that humans and wildlife shared exposures. Moreover, it emphasizes the importance of suspecting disease in the other when disease is found in either human or wildlife populations.     

Prevalence of Neospora caninum antibodies and factors associated with their presence in dairy cattle of the north of Paraná state, Brazil

To determine the prevalence of Neospora caninum antibodies and associated factors, blood sera from 623 female dairy cattle from 23 farms in the north of the state of Paraná, Brazil, were analyzed by means of the indirect immunofluorescent-antibody test (IFAT > or = 25). Serum samples from 134 dogs living on the same farms also were tested for N. caninum antibodies (IFAT > or = 50), and the presence of dogs was associated with the prevalence of N. caninum antibodies in cattle. The overall seroprevalence in cattle was 14.3%, mainly in animals over 24 months of age. Seroprevalence in Holsteins (15.1% of 558) was greater than in mixed-breed cattle (7.7% of 65). Age (> or =24 months) of cattle, feeding silage and/or concentrate produced on the farm were associated; antibodies were found in 21.6% of dogs; and the presence of dogs was associated with the prevalence of N. caninum antibodies in cattle.     

Conjugation and Characterization of Latex Particles with Toxoplasma gondii–specific Immunoglobulin Y Antibodies for Diagnostic Aim and Evaluation Efficiency in In Vitro Culture

Toxoplasma gondii is a parasite that causes severe health problems in the world. Toxoplasmosis, an infection caused by T. gondii, leads to high risk of mortality in patients with immunodeficiency, transplantation, and cancer. Besides that, it causes miscarriages in pregnancy, various abnormalities such as hydrocephalus in infants and congenital diseases. Because the clinical indication of the disease is not specific, it is confused with many diseases, and this leads to the necessity of directly detecting the presence of the toxoplasmosis. Therefore, various diagnostic assays are needed for the diagnosis of the disease. Amongs them, latex agglutination assay is widely used for the detection of specific antibodies or antigens in samples. Latex particles are coated with immunogenic molecules (antigens) to detect antibodies in the blood or used to identify antigens when coated with specific antibodies. In both, aggregation of latex particles results in agglutination. Monoclonal antibodies are often used in latex agglutination assay as in other diagnostic methods. However, monoclonal antibodies can be produced in low quantities at a high cost. Besides, to produce monoclonal antibodies, an experienced staff, a well-equipped cell culture laboratory, a long period of time, and a burdened budget are needed. In recent years, as an alternative to monoclonal antibodies, immunoglobulin Y (IgY) antibodies, which are obtained from chicken eggs, and specifically produced against desired antigenic constructs, have become quite attractive in terms of both low cost and abundant production without requiring infrastructure. In contrast, the latex assay based on IgY antibodies for use in the diagnosis of T. gondii has not been developed. This study aimed to conjugate T. gondii-specific IgY antibodies to latex particles, characterize the particles by Fourier transform infrared spectroscopy, scanning electron microscopy, and spectroscopic methods, and finally demonstrate the interaction with T.gondii parasites in culture with scanning electron microscopy analysis.     

Developing vaccines to control protozoan parasites in ruminants: dead or alive?

Protozoan parasites are among some of the most successful organisms worldwide, being able to live and multiply within a very wide range of hosts. The diseases caused by these parasites cause significant production losses in the livestock sector involving reproductive failure, impaired weight gain, contaminated meat, reduced milk yields and in severe cases, loss of the animal. In addition, some protozoan parasites affecting livestock such as Toxoplasma gondii and Cryptosporidium parvum may also be transmitted to humans where they can cause serious disease. Data derived from experimental models of infection in ruminant species enables the study of the interactions between parasite and host. How the parasite initiates infection, becomes established and multiplies within the host and the critical pathways that may lead to a disease outcome are all important to enable the rational design of appropriate intervention strategies. Once the parasites invade the hosts they induce both innate and adaptive immune responses and the induction and function of these immune responses are critical in determining the outcome of the infection. Vaccines offer green solutions to control disease as they are sustainable, reducing reliance on pharmacological drugs and pesticides. The use of vaccines has multiple benefits such as improving animal health and welfare by controlling animal infections and infestations; improving public health by controlling zoonoses and food borne pathogens in animals; solving problems associated with resistance to acaricides, antibiotics and anthelmintics; keeping animals and the environment free of chemical residues and maintaining biodiversity. All of these attributes should lead to improved sustainability of animal production and economic benefit. Using different protozoan parasitic diseases as examples this paper will discuss various approaches used to develop vaccines to protect against disease in livestock and discuss the relative merits of using live versus killed vaccine preparations. A range of different vaccination targets and strategies will be discussed to help protect against: acute disease, congenital infection and abortion, persistence of zoonotic pathogens in tissues of food animals and passive transfer of immunity to neonates.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in goats slaughtered in the public slaughterhouse of Patos city, Paraíba State, Northeast region of Brazil

The prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies was investigated in goats slaughtered in the public slaughterhouse of Patos, State of Paraíba, Northeast region of Brazil, and possible associations between sex of the animals and antibody prevalence were verified. Three-hundred six blood samples from goats collected before slaughter by jugular venopuncture were used. For the serologic diagnosis of T. gondii and N. caninum, the indirect fluorescent-antibody test (IFAT) with cut-off values 64 and 50, respectively, was carried out. The prevalence of anti-T. gondii antibodies was 24.5% [95% CI=19.8-29.7%] with titers ranging from 64 to 4096, and anti-N. caninum antibodies was 3.3% (95% CI=1.6-5.9%) with titers ranging from 50 to 400. There were no associations between sex of animals and prevalence of anti-T. gondii and anti-N. caninum antibodies.