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).     

Anti-neosporal IgG and IgE antibodies in canine neosporosis

Neospora caninum infection provokes neurological disorders, recurrent abortion and death in dogs and cattle. Dogs are both intermediate and definitive host of N. caninum. Thus, the development of sensitive and specific immunoassays to diagnose canine neosporosis is essential to control this disease. This work investigated serum anti-neosporal IgG and IgE antibodies in 140 dogs represented by 30 healthy animals (group I), 11 dogs showing acute N. caninum infection (group II), 50 urban dogs with serological evidence of canine neosporosis in indirect fluorescent antibody test (IFAT) (group III) and 49 urban dogs without clinical and laboratory evidences of neosporosis (group IV). Enzyme-linked immunosorbent assay (ELISA) and western immunoblotting, both using a soluble N. caninum tachyzoite antigen (SNA), investigated these two isotypes of antibodies, while a Urea-ELISA measured the avidity of the IgG antibodies. Anti-Toxoplasma gondii IgG antibodies were also investigated in the animals. Anti-neosporal IgG was found in all animals from groups II and III, whereas 32.7% (16/49) of dogs from group IV were reactive. IgG antibodies of low avidity were demonstrated in dogs from group II (median 35.3%), while animals from groups III and IV had IgG antibodies of high avidity (medians of 61.5% and 61.7% respectively). IgE antibodies were found in four (13.3%) and five (16.6%) dogs from groups III and IV respectively. Dogs presenting acute infection (group II) or chronic infection (group III) had IgG antibodies to several neosporal antigens, mainly of 29-30 and 35 kDa, while 13 of 16 dogs from group IV recognized antigens from 14 to 170 kDa. Antibodies to T. gondii were detected in 36 of 50 (72%) sera from group III and 25 of 49 (51%) sera from group IV. We concluded that IgG-ELISA and Urea-ELISA with SNA may substitute for IFAT in both laboratory routine and epidemiological studies of canine neosporosis.     

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.     

Genotypic characterization of Toxoplasma gondii in sheep from Brazilian slaughterhouses: new atypical genotypes and the clonal type II strain identified

Toxoplasma gondii strains are genetically diverse in South America. To date, hundreds of T. gondii isolates from different animal hosts were genotyped in Brazil, most of them are different from those identified around the world. This study aimed to determine T. gondii infection rate in sheep from Brazilian slaughterhouses, as well as the genotype of these isolates. T. gondii antibodies were detected in 66/602 (10.96%) serum samples through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT). MAT-HS and IFAT-IgG presented high concordance (0.95) and strong correlation (r=0.79). T. gondii DNA was detected in tissue samples of 33% (22/66) serum positive sheep by PCR of the 529 bp repetitive element. In the bioassay in mice, T. gondii were detected in mice brain or muscle tissues in 30% (20/66) of serum positive sheep. Positive samples were typed through Restriction Fragment Length Polymorphism (RFLP-PCR) using 11 markers: SAG1, SAG2 (5'-3'SAG2 and alt.SAG2), SAG3, BTUB, GRA6, L358, c22-8, c29-6, PK1, Apico and CS3. Of 22 samples, 13 were positive and 9 genotypes were identified. Four of these 9 genotypes are unique. Nine samples had negative results in RFLP-PCR typing, which may be due to low DNA concentration. Six isolates were virulent killing mice between 12 and 25 days postinfection. Two non-virulent isolates belonged to clonal type II genotype, which were not observed in Brazil previously. These findings confirm the high diversity and high frequency of virulent genotypes among Brazilian animals. This study also proved the presence of type II T. gondii in Brazil.     

Seroprevalence of Toxoplasma gondii infection in goats and sheep in Zimbabwe

Seroprevalence rates of Toxoplasma gondii anti-antibodies in adult goats and sheep from different parts of Zimbabwe were determined. A total of 225 (67.9%) of the 335 serum samples tested were positive for anti-T. gondii IgG antibodies with the indirect fluorescent antibody test. There were differences in antibody seroprevalences among communal land goats from the different agro-ecological zones (Natural regions llb and III: 80 and 96.7%, respectively; Natural region IV: 65.9%; Natural region V: 45%; and Natural region III had a significantly higher seroprevalence than IV and V. The highest seroprevalences found in Natural regions II b and Ill are likely to be linked to the existence of more households and hence the possibility of a higher concentration of domestic cats that increases the chances of environmental contamination with their faeces harbouring T. gondii oocysts. The seroprevalence rate in sheep from a large commercial farm (10%) was significantly lower than that of sheep reared under the communal grazing system (80%). Overall, significantly higher proportions of seropositive animals had antibody titres of 1:50 (34.2% of 225) and 1:100 (44% of 225) as compared to the 9.8% and 12% with antibody titres of 1:200 and > or =1:400, respectively.     

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.     

The prevalence and avidity of Toxoplasma gondii IgG antibodies in pigs from Brazil and Peru

Raw or inadequately cooked pork is an important source of Toxoplasma gondii infection, and the infection rate in animals used as human food, is an important risk predictor. The prevalence of this infection was estimated in 396 sera from 5-month old pigs obtained at abattoirs in S?o Paulo, Brazil (300) and Lima, Peru (96). The seroprevalence was higher in pigs from Peru (32.3%) as compared to Brazil (9.6%), as detected by ELISA and Western blot. Hemagglutination gave poor resolution which was not useful for the diagnosis of T. gondii infection. Specific antibody avidity is correlated with infection time, as shown in experimentally infected piglets. Using an arbitrary cut-off of 50% avidity index, Brazilian pigs were found to be more recently infected than Peruvian pigs. Pork should be considered a significant source of human T. gondii infection both in Brazil and Peru. Avidity assays could help in the detection of the time of T. gondii infection in pigs, allowing preventive management.     

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.     

Toxoplasma gondii in sheep from the Campania region (Italy)

A cross-sectional serological survey was conducted in order to evaluate, irrespective of abortion, the Toxoplasma gondii infection in pastured sheep from the Campania region of southern Italy. A geographical information system was used in order to uniformly sample the ovine farms (n=117) throughout the entire region. Blood and milk samples were collected from 10 adult sheep (>18 months) on each farm (total number=1170 sheep). Serum samples were tested for the presence of IgG antibodies to T. gondii using a commercial indirect fluorescent antibody test. For each farm, the 10 milk samples collected were pooled in order to obtain a single milk sample per farm (total number=117 milk samples). The 77.8% (91/117) of the farms and the 28.5% (333/11,170) of the sheep resulted positive by serology. In addition, the presence of T. gondii DNA was detected by PCR in 4 milk samples out of the 117 examined (3.4%).     

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.     

Prevalence and risk factors for anti-Toxoplasma gondii antibodies in goats of the Seridó Oriental microregion, Rio Grande do Norte state, Northeast region of Brazil

The prevalence and risk factors for anti-Toxoplasma gondii antibodies were investigated in goats of the Seridó Oriental microregion, Rio Grande do Norte state, Northeast region of Brazil. Three hundred and sixty-six blood samples from goats collected by jugular venopuncture were used. For the serologic diagnosis of Toxoplasma gondii infection, the indirect fluorescent-antibody test (IFAT) with cut-off value 1:64 was carried out. The prevalence of anti-T. gondii antibodies was 30.6% [95% CI=25.9-35.6%] with titers ranging from 1:64 to 1:16,384. The multivariate logistic regression analysis showed that the risk factors associated to anti-T. gondii antibodies were presence of cats in the herd, extensive/semi-intensive management systems and lack of mineral supplementation.     

Survey of ovine and caprine toxoplasmosis by IFAT and PCR assays in Sardinia, Italy

During the period 1999-2002, we have analyzed 9639 serum samples and 815 aborted samples (670 fetuses and 145 placenta) from 964 ovine and caprine farms distributed over all Sardinia island. After abortion notification, sera collected at random from adult animals were examined to detect simultaneously IgG and IgM antibodies specific to Toxoplasma gondii by indirect immunofluorescence assay, whereas fetuses and placenta were analyzed by a single tube nested PCR assay. Specific IgG antibodies were detected in 2048 (28.4%) sheep and 302 (12.3%) goats, specific IgM antibodies were found in 652 (9%) sheep and 139 (5.6%) goats. From a total of 2471 ovine and 362 caprine fetal samples including muscle, liver, abomasum, spleen, brain and placenta, 271 (11.1%) ovine and 23 (6.4%) caprine samples were T. gondii PCR-positive. Although T. gondii DNA was amplified from different types of tissues, placenta was the tissue with the highest detection rate. On the one hand, these results indicate that the seroprevalence of T. gondii infection in sheep and goats is relatively high, on the other PCR results demonstrate that T. gondii has a significant role in ovine and caprine abortion. Adequate management might be useful and essential to control the toxoplasmosis in the sheep and goats herds of Sardinia.     

龙岩市部分地区牛、羊弓形虫病的血清学调查

目的：采集龙岩市部分地区牛、羊血清,检测福建省牛和羊弓形虫病的感染情况。方法：应用酶联免疫吸附试验检测牛和羊血清弓形虫IgG抗体。结果：49份牛血清弓形虫IgG抗体均呈阴性;35份羊血清中,8份弓形虫IgG抗体呈阳性,阳性率为22．86％（8／35）。结论：龙岩市部分地区羊弓形虫的感染率较高,应引起人们重视。     

Comparison of a commercial ELISA with the modified agglutination test for the detection of Toxoplasma gondii infection in naturally exposed sheep

Surprisingly few commercial ELISAs are available for the detection of Toxoplasma gondii infection in animals, and none for use in sheep have been evaluated. We thus compared the Bommeli Diagnostics ELISA Toxotest for the detection of T. gondii antibodies in ruminants with the reference modified agglutination test (MAT) in a series of 180 sheep sera. ELISA results were analysed at two cut-off levels (30%, comprising both weakly positive and positive results, and 100%, comprising only positive results), and compared with MAT at three cut-off levels (titre of 1 : 25, 1 : 50 and 1 : 100). The results showed a moderate agreement of ELISA at both cut-offs (kappa = 0.46 and 0.51) with MAT at a cut-off titre of 1 : 100. However, the specificity and positive predictive value were above 95% only at an ELISA cut-off of 100%, indicating its potential as a diagnostic test, particularly in areas with a high prevalence of infection. On the other hand, lower sensitivity and negative predictive value limit its value as a screening test. Thus, the ELISA Toxotest may be used for quick diagnosis of T. gondii infection in sheep in the field, i.e. for the differential diagnosis of ovine abortion storms.     

我国部分地区牛羊弓形虫血清流行病学调查

为了解我国牛羊弓形虫病流行情况,应用间接血凝试验（IHA）对河南、山东、山西、内蒙古、云南、贵州6省区151份牛血清、50份奶样、490份羊血清进行了弓形虫病血清流行病学调查。结果显示：151份被检牛血清和50份牛奶样品,弓形虫抗体均为阴性。490份羊血清弓形虫抗体总阳性率5.71％,其中母羊、公羊血清阳性率分别为4.03％和9.79％;山羊、绵羊、杂交羊血清阳性率分别为6.58％、4.81％和5.13％;阳性率最高的为公山羊（13.2％）,最低的为母绵羊（2.96％）。28份阳性羊血清中,75％的抗体滴度为1：64,25％的抗体滴度为1：256。1岁后的羊,随年龄增长,血清阳性率升高。     

Toxoplasma gondii in Switzerland: a serosurvey based on meat juice analysis of slaughtered pigs, wild boar, sheep and cattle

Toxoplasmosis is one of the most important zoonotic diseases worldwide and is caused by the protozoan Toxoplasma gondii. Besides vertical infection during pregnancy, humans can get infected post-natally either by peroral uptake of sporulated Toxoplasma oocysts or by ingestion of tissue cysts upon consumption of raw or undercooked meat. The aim of this study was to approximate the risk of human infection via meat consumption by estimating the seroprevalence of T. gondii in slaughtered animals in Switzerland and to compare data with prevalences assessed 10 years ago. The study included pigs, cattle, sheep and wild boar of different age groups and housing conditions whenever possible and applicable. A P-30-ELISA was used to detect T. gondii-specific antibodies and to determine seroprevalences in meat juice of slaughtered animals. A total of 270 domestic pigs (120 adults, 50 finishing, 100 free-ranging animals), 150 wild boars, 250 sheep (150 adults, 100 lambs) and 406 cattle (47 calves, 129 heifers, 100 bulls, 130 adult cows) were tested. Seropositivity increased with the age of the assessed animals. Independent of the age-group, the overall seroprevalence was lowest in wild boars (6.7%), followed by pigs (23.3%), cattle (45.6%) and sheep (61.6%), respectively. Conventional fattening pigs and free-ranging pigs surprisingly had comparable seroprevalences (14.0% and 13.0%, respectively). Unlike in other European countries, where generally a decrease in the number of seropositive animals had been observed, we found that the prevalence of seropositive animals, when compared with that of 10 years ago, had increased for most species/age groups. Conclusively, the results demonstrated a high seroprevalence of T. gondii in animals slaughtered for meat production and revealed that increasing age of the animals is a more important risk factor than housing conditions in Switzerland.     

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.     

Seroprevalence of T. gondii in sheep from Marrakech, Morocco

Toxoplasma gondii is a ubiquitous intracellular protozoan parasite transmitted by food. Concerning this parasite, there are few studies done in Morocco. In this study, 261 sera from sheep intended for consumption in Marrakech were subjected to the Toxoplasma ELISA based serology test for the detection of anti-T. gondii specific IgG confirming a past infection. Of the total tested 72 (27.6%) sera were positive for IgG. This result shows that the seroprevalence approaches the world average and is similar to what is found in other cities of Morocco. This has prompted us to investigate other animal species in the region in order to evaluate the degree of contamination by this parasite as well as the potential risk incurred on consumption of their meat.     

Seroepidemiology of Neospora caninum and Toxoplasma gondii infection in yaks (Bos grunniens) in Qinghai, China

Neospora caninum is a protozoan parasite and is closely related to Toxoplasma gondii, but they are antigenically different. N. caninum and T. gondii infection in a variety of animals such as cattle, dogs, and cats has been reported, but there is little information on the infection of these parasites in domestic yaks. Seroprevalence of antibodies to T. gondii and N. caninum in yaks (Bos grunniens) from eight regions of Qinghai, China were investigated by the indirect agglutination test (IAT) and ELISA, respectively. A total of 112 (11.8%) of 946 serum samples were positive for antibodies to T. gondii, and 21 samples (2.2%) were positive to N. caninum. Two of the yaks had antibodies to both parasites. There was no apparent association of T. gondii infection with age of the animals. The results indicate that T. gondii infection is prevalent in Chinese yaks in most parts of Qinghai province and N. caninum infection rate in the same species is relatively low. This is the first large study showing the infection of T. gondii and N. caninum in domestic yaks.     

屠宰羊弓形虫感染情况调查

为了保障羊肉制品的生物安全,对屠宰羊进行弓形虫感染情况调查以及对羊肉进行弓形虫检测。采用间接血凝试验,对677份屠宰羊的血清进行了弓形虫抗体检测。结果显示,弓形虫抗体阳性有47份,阳性率为6.9%。