First isolation and molecular characterization of Toxoplasma gondii from finishing pigs from São Paulo State, Brazil

Toxoplasma gondii infection is widely prevalent in humans in Brazil. Among the food animals, pigs are considered the most important meat source of T. gondii for infection in humans. In the present study, we report the first isolation of viable T. gondii from finishing pigs in Brazil. Antibodies to T. gondii were found in 49 (17%) of 286 pigs prior slaughter using the modified agglutination test (MAT) at a serum dilution of 1:25. Attempts were made to isolate T. gondii from 28 seropositive pigs. Samples of heart, brain, and tongue from each pig were pooled, digested in acid pepsin, and bioassayed in five mice per pig. Viable T. gondii was isolated from seven pigs; all isolates were lethal for mice. Restriction fragment length polymorphism on products of SAG2 locus amplified by PCR revealed that two isolates were Type I and five were Type III. The results indicate that phenotypically and genetically T. gondii isolates from pigs from Brazil are distinct from isolates of T. gondii from pigs in the USA.     

Toxoplasma gondii infection in slaughter pigs in Serbia: seroprevalence and demonstration of parasites in blood

ABSTRACT: A seroepizootiological study of Toxoplasma gondii infection involving a total of 488 slaughter pigs (468 market-weight pigs and 20 sows) in the Belgrade area, also included examination of the presence of T. gondii in the blood. Blood sampled at the slaughter line was examined for specific antibodies by modified direct agglutination, and blood clots of those seropositive at titres of 1:50-1:12800 were bioassayed in mice. The overall seroprevalence was 9.2%, significantly higher (p = 0.0063) in sows (30.0%) than in market-weight pigs (8.3%). Amongst the 22 bioassays performed, a total of 16 (72.7%) were positive, by observation of T. gondii cysts (12), seropositivity (7, including 3 in which cysts were not detected), and/or detection of T. gondii DNA by real-time PCR (12, including one otherwise negative). The positive bioassays originated from the blood of 12 market-weight pigs and 4 sows. Despite a general increase in the rate of demonstration of T. gondii with the increase in the specific antibody level, the association was not significant (p = 0.101). The risk of infection was 41-fold increased in sows vs market-weight pigs, and 15-fold in pigs from smallholders' finishing type farms vs those from large farrow-to-finish farms. The presence of viable T. gondii in a proportion of the samples indicates that some of the pigs had an active parasitaemia at the time of slaughter, which, along with the seroprevalence established, points to a potential source of human infection in Serbia. This is the first report on parasitaemia in naturally infected swine.     

High prevalence and genotypes of Toxoplasma gondii isolated from organic pigs in northern USA

The ingestion of undercooked pork infected with Toxoplasma gondii is considered an important source of transmission of this parasite. While T. gondii infection in confinement raised market pigs (market pigs are typically used for fresh, unprocessed pork products) in the USA has decreased significantly over the last 20 years, infection levels in pigs with access to the outdoors can be quite high. An upsurge in consumer demand for 'organically raised', 'humanely raised' and 'free range' pork products has resulted in increasing numbers of hogs being raised in non-confinement systems. To determine T. gondii infection rate in these organic pigs, prevalence of T. gondii in organically raised pigs in two establishments (Farm 1, Farm 2) in Michigan was investigated. Serum and tissue samples from 33 pigs on the farm were available for T. gondii evaluation at slaughter. Serological testing was performed using both ELISA and the modified agglutination test (MAT). Antibodies to T. gondii were detected by both ELISA and MAT in 30 of 33 animals with MAT titers of 1:25 in three, 1:50 in six, 1:100 in seven, 1:200 in 13, and 1:400 in one. Hearts of all 33 pigs were bioassayed for T. gondii in mice; T. gondii was isolated from 17 pigs including one from a seronegative (both ELISA and MAT) pig. Genetic typing of 16 of the 17 T. gondii isolates using the SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico loci revealed clonal Type II from Farm 1 and clonal Type III on Farm 2. These results revealed very high prevalence of T. gondii in organic pigs for the first time in USA, indicating potentially increased health risk of consuming organic swine products.     

Seroprevalence and Toxoplasma gondii-IgG avidity in sheep from Lajes, Brazil

Sheep are important domestic animals in the Northeast region of Brazil due to their minimal rearing and maintenance costs, and to their production of both meat and milk. In animals, Toxoplasma gondii infection results in significant reproductive and economic losses. The epidemiology of toxoplasmosis in sheep in the Northeast of Brazil has been little studied; particularly in Rio Grande do Norte State. Sera from 102 sheep intended for consumption in Lajes were subjected to the Toxoplasma-ELISA test to detect anti-T. gondii specific IgG confirming a past infection. Of the total tested, 30 (29.41%) sera were positive for IgG with an increasing number of positive animals with advancing age. We used IgG avidity ELISA in 30 positive samples and observed that 6 (20%) had low avidity antibodies and 24 (80%) had high avidity antibodies. Epidemiological studies are required in order to identify sources of infection for these hosts as well as their impact on animal breeding in the region and risk of transmission to humans.     

Soil contamination of Toxoplasma gondii oocysts in pig farms in central China

Toxoplasmosis in pigs is a large threat to pig industry as well as pork consumers. Most pigs become infected by ingestion of oocysts from contaminated environment (soil, water and feed) or infected animal tissues postnatally. In the present study, field studies were conducted to evaluate the relationship between soil contamination status of Toxoplasma gondii oocysts and T. gondii infection in pigs in 12 pig farms with different density of cats in central China. The presence of T. gondii oocysts in soil were determined by PCR and loop-mediated isothermal amplification (LAMP). T. gondii DNA was found in 11 farms with different cat density excepting one farm exposed to low cat density. Twenty (21.1%) and 36 (37.9%) of 95 soil samples were T. gondii positive by PCR and LAMP, respectively (0.01相似文献   

Comparison of detection methods for Toxoplasma gondii in naturally and experimentally infected swine

Results from recent serological surveys and epidemiological studies show that pigs raised in a variety of management systems can be carriers of the tissue cyst stage of Toxoplasma gondi. This parasite can be transmitted to humans through the consumption of improperly prepared pork, making detection and removal of infected swine carcasses from the food chain an important food safety issue. Several methods are available for detection of T. gondii infected swine, including serological assays, polymerase chain reaction, and animal bioassays. The aim of the present study was to compare the detection sensitivities of six of these commonly used methods for detection of T. gondii infection in tissues from naturally and experimentally infected pigs. The results indicate that a serum-based ELISA is the most sensitive method, of those tested, for detection of T. gondii infected swine.     

几种检测猪弓形虫病ELISA诊断试剂盒的对比

以人工感染弓形虫的猪阳性血清及感染前的阴性血清为样品,比较3个国产(A、B、C)和2个国外(D、E)ELISA试剂盒检测猪弓形虫病的敏感性。用这5种ELISA试剂盒,检测人工感染弓形虫的猪阳性血清42份及感染前的阴性血清45份,并参考PCR扩增结果,比较这5种ELISA试剂盒检测猪弓形虫病的敏感性。5种试剂盒中,1种国产试剂盒的检测结果与PCR结果完全一致,其余4种试剂盒的检测结果都与PCR检测结果差别较大。结果表明,国产试剂盒A的检测效果最佳,国外试剂盒与部分国内试剂盒的检测结果相当,临床选用试剂盒时,可选择检测效果最佳的试剂盒A,不必盲目选择国外试剂盒。     

Detection of specific antibodies to Neospora caninum and Toxoplasma gondii in naturally infected alpacas (Lama pacos), llamas (Lama glama) and vicunas (Lama vicugna) from Peru and Germany

Sera of an experimentally Neospora caninum infected llama and a non-infected control llama were used to establish an immunoblot, an ELISA and an IFAT to detect antibodies against N. caninum tachyzoites. Subsequently, serum samples collected from a total of 871 South American Camelids (SAC: Lama glama, Lama pacos, Lama vicugna) of two farms in Peru and from 32 SAC of a farm in central Germany were examined for antibodies against N. caninum and Toxoplasma gondii. Based on the recognition of specific bands in the immunoblot, sera of SAC from Peru were differentiated into N. caninum-positive (n = 18) and T. gondii-positive (n = 30) samples and into samples negative or inconclusive for both parasites. Using the immunoblot results as the reference, a modified version of the p38-ELISA and the IFAT were evaluated for detecting N. caninum antibodies in SAC sera. Applying a cut-off as determined by two graph-receiver operating characteristic analysis both, the ELISA and the IFAT, exhibited a sensitivity and specificity of about 95% in the SAC sera from Peru. Serological testing confirmed that SAC may become infected with N. caninum under field conditions in Peru. In addition to alpacas and llamas also 114 wild living vicunas had been examined for antibodies against N. caninum. However, only the alpacas and llamas but no vicunas were found N. caninum-positive. In contrast, T. gondii-seropositive animals were detected in all three SAC species. The lack of N. caninum-seropositive vicunas indicates that in the study area in Peru wild canids might not serve as definitive hosts of N. caninum while for T. gondii a life cycle including wild felids is likely. On the German farm no N. caninum- but only T. gondii-seropositive SAC (n = 14) were detected. The seroprevalence of T. gondii infection was significantly higher in adult SAC (alpacas in Peru, llamas in Germany) than in crias (i.e. < 12 months old foals) indicating that the predominant route of infection is post natal. Since the present study was restricted to a few farms, the seroprevalences determined are not representative. However, our results confirm natural infections with N. caninum and T. gondii in SAC. Whether these infections are linked to any disease, e.g. reproductive losses, has to be clarified in further studies.     

Evaluation of an in-house TgSAG1 (P30) IgG ELISA for diagnosis of naturally acquired Toxoplasma gondii infection in pigs

Toxoplasma gondii is an apicomplexan protozoan parasite which is able to infect a large variety of warm-blooded animals. Raw or undercooked pork has been regarded as an important source of infection for humans. The aim of this study was to evaluate an in-house enzyme-linked immunosorbent assay to diagnose natural T. gondii infection in swine using native affinity chromatography-purified T. gondii surface protein-1 (TgSAG1-ELISA) as antigen, comparing its performance to that of indirect fluorescent antibody test (IFAT) and immunoblotting (IB). To obtain a panel of sera showing the evolution of the antibody response in the time course 12 pigs were experimentally inoculated intravenously (iv) with tachyzoites of the T. gondii strains RH (clonal type I), ME49 (clonal type II) and NED (clonal type III) and serologically monitored for a period of 11 weeks. Both IFAT and ELISA showed a similar time course of antibody response to T. gondii; but by IFAT this response was characterized by rapidly rising titers with peaks at two weeks post inoculation (wpi), while the ELISA indices increased slowly and reached a maximum in most animals at five wpi. Three-hundred randomly selected sera from a total of 602 pigs of different ages derived from outdoor and indoor farms from Argentina were analyzed. Serum samples testing either positive or negative by both IFAT and IB were considered as "relative standards of comparison" (RSC). Sensitivity and specificity of TgSAG1-ELISA were obtained by a Receiver Operating Characteristics (ROC) analysis and statistical agreement among serological tests was evaluated. Antibodies to T. gondii were detected in 160 of 300 sera (53.3%) by IB, in 133 of 300 (44.3%) by IFAT and in 123 of 300 sera (41%) by TgSAG1-ELISA. One hundred and eleven sera tested positive and 118 sera tested negative by both IFAT and IB (RSC); 103 of 111 positive RSC sera tested positive by TgSAG1-ELISA, and 116 of 118 negative RSC sera tested negative by TgSAG1-ELISA. Agreement observed between RSC and TgSAG1-ELISA was almost perfect (κ=0.9124, p≥0.05) and between IFAT and IB was moderate (κ=0.53, p≥0.05). Relative sensitivity and specificity of the TgSAG1-ELISA using a cut-off index of 0.204 were of 92.8% and 98.3%, respectively. ROC analysis revealed that TgSAG1-ELISA was highly accurate (AUC=0.983) relative to the RSC. According to the results in this study, the ELISA based on affinity purified T. gondii surface antigen TgSAG1 was useful for the specific and sensitive detection of antibodies to this protozoan parasite in naturally infected pigs.     

Detection of T. gondii in tissues of sheep and cattle following oral infection.

It has been reported in the literature that cattle are more resistant to toxoplasmosis than sheep. Congenital disease due to T. gondii infection is rarely reported in cattle whereas the parasite is a major cause of abortion and neonatal mortality in sheep. It is believed that sheep remain chronically infected for life. Undercooked meat from infected sheep is an important source of infection for man. In contrast cattle are thought to harbour fewer parasite tissue cysts which may not persist for the lifetime of the host. Therefore, cattle are believed to pose less of a risk for human infection. In this study we examined the presence of T. gondii within a range of tissues in sheep and cattle at 6 weeks and 6 months following oral infection with 10(3) or 10(5) sporulated oocysts of T. gondii. The presence of parasite was determined by bioassay in mice and using polymerase chain reaction (PCR). The results from this study show that T. gondii was more frequently and consistently detected in sheep, in particular within brain and heart tissues, whereas parasites were not detected in the samples of tissues taken from cattle. T. gondii was more frequently detected in sheep given the higher dose of T. gondii. Examination of tissues at either 6 weeks or 6 months after infection did not appear to affect the distribution of T. gondii. The polymerase chain reaction has more specificity and sensitivity when detecting the presence of T. gondii in large animals than histological detection.     

The occurrence of Toxoplasma gondii and Neospora caninum as regards meat hygiene

Toxoplasma gondii and Neospora caninum are phenotypically and phylogenetically closely related cyst-forming coccidia, both of which may cause abortion in livestock animals. T. gondii exhibits also zoonotic potential by causing diaplacental infections in the human fetus and harmful infections in immunosuppressed individuals. Humans get infected either by consuming inappropriately prepared cyst-containing meat or by ingesting oocysts originating from cat feces. Therefore, in order to assess infection risk we need to have knowledge on the prevalence of the parasite in consumable meat and thus slaughtered animals. So far, no data indicate any zoonotic potential for N. caninum. Due to its high economic impact in the bovine production in Switzerland, we included this parasite in the present study as well. The prevalence of both parasite species were investigated by PCR in muscle and brain samples of slaughtered bovines, sheep, pigs and horses. Comparatively, a serum sample from each animal was simultaneously tested serologically by a Toxoplasma-P30-ELISA and a Neospora-SA-ELISA. The prevalences determined by the T. gondii-PCR were the followings: adult cows 3%, young bulls 2%, young cows prior to gravidity 6%, calves 1%, sheep 6%, horses and pigs each 0%. For N. caninum, the PCR-prevalence was 2% for adult cows and 0% for all other animal groups. Conversely, the seroprevalences were much higher for both parasite species and all animal groups, with the exception of the fattening pigs. However, as T. gondii was principally detectable in bovine (cows and calves) as well as in sheep meat, the consumption of this meat harbours a potential infection risk for humans. In contrast, the lack of any parasite detectability in fattening pig and horse meat allows to consider this infection source as neglectable when compared to bovine and ovine meat.     

Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland

The protozoan parasite Toxoplasma gondii infects almost all warm blooded animal species including humans, and is one of the most prevalent zoonotic parasites worldwide. Post-natal infection in humans is acquired through oral uptake of sporulated T. gondii oocysts or by ingestion of parasite tissue cysts upon consumption of raw or undercooked meat. This study was undertaken to determine the prevalence of oocyst-shedding by cats and to assess the level of infection with T. gondii in meat-producing animals in Switzerland via detection of genomic DNA (gDNA) in muscle samples. In total, 252 cats (44 stray cats, 171 pet cats, 37 cats with gastrointestinal disorders) were analysed coproscopically, and subsequently species-specific identification of T. gondii oocysts was achieved by Polymerase Chain Reaction (PCR). Furthermore, diaphragm samples of 270 domestic pigs (120 adults, 50 finishing, and 100 free-range animals), 150 wild boar, 250 sheep (150 adults and 100 lambs) and 406 cattle (47 calves, 129 heifers, 100 bulls, and 130 adult cows) were investigated by T. gondii-specific real-time PCR. For the first time in Switzerland, PCR-positive samples were subsequently genotyped using nine PCR-restriction fragment length polymorphism (PCR-RFLP) loci (SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) for analysis. Only one of the cats shed T. gondii oocysts, corresponding to a T. gondii prevalence of 0.4% (95% CI: 0.0-2.2%). In meat-producing animals, gDNA prevalence was lowest in wild boar (0.7%; 95% CI: 0.0-3.7%), followed by sheep (2.0%; 95% CI: 0.1-4.6%) and pigs (2.2%; 95% CI: 0.8-4.8%). The highest prevalence was found in cattle (4.7%; 95% CI: 2.8-7.2%), mainly due to the high prevalence of 29.8% in young calves. With regard to housing conditions, conventional fattening pigs and free-range pigs surprisingly exhibited the same prevalence (2.0%; 95% CI: 0.2-7.0%). Genotyping of oocysts shed by the cat showed T. gondii with clonal Type II alleles and the Apico I allele. T. gondii with clonal Type II alleles were also predominantly observed in sheep, while T. gondii with mixed or atypical allele combinations were very rare in sheep. In pigs and cattle however, genotyping of T. gondii was often incomplete. These findings suggested that cattle in Switzerland might be infected with Toxoplasma of the clonal Types I or III, atypical T. gondii or more than one clonal Type.     

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.     

A computer simulation of the prevention of the transmission of Toxoplasma gondii on swine farms using a feline T. gondii vaccine

Transmission of Toxoplasma gondii on swine farms was investigated using a deterministic dynamic computer simulation model. A primary focus was to evaluate a feline T. gondii vaccine. Animal populations (swine and cats) were compartmentalized based on the stage of T. gondii infection. Simulations were run under conditions of closed and equilibrium population size. Model parameters were varied in a factorial experimental design to test the following hypotheses: T. gondii infection in finishing pigs decreases with (1) vaccination of susceptible cats, (2) an increase in the proportion of cats captured for vaccination, (3) a decrease in the initial number of cats, (4) a decrease in the initial T. gondii prevalence in cats and (5) a decrease in oocyst-survival time. Seeding conditions included a total of 10, 20, 30, 40 or 50 cats, initial T. gondii prevalences in cats of 30, 60 or 90%, vaccination of 0, 50 or 75% of the cats and two vaccination schedules (the field schedule from a prior trial and a weaning-vaccination schedule). Simulations were run at oocyst-survival times of 52, 39 and 26 weeks. T. gondii prevalence in finishing pigs was recorded every week for 10 years. The probability of elimination of T. gondii from finishing pigs increased with a decrease in the number of cats and a decrease in oocyst-survival time.The last-year average prevalence was used as the outcome in a multiple linear regression analysis. Decreased T. gondii prevalence in finishing pigs was the result of a decrease in the initial number of cats on the farm (squared semipartial correlation coefficient (sr(2))=47%), decreased oocyst survival (sr(2)=35%), using the weaning-vaccination schedule (sr(2)=7%) and vaccination versus non-vaccination (sr(2)=5%). Unexpectedly, the initial T. gondii prevalence in cats had no effect on T. gondii prevalence in finishing pigs. The simulation supports the field trial indicating vaccine effectiveness. However, vaccination had less impact on decreasing T. gondii infection in finishing pigs than a decrease in the number of farm cats.     

Longitudinal study on the seroprevalence of Toxoplasma gondii infection in four German pig breeding and raising farms

There are very few current data on the prevalence of Toxoplasma (T.) gondii in German pig farms. Consequently a reliable risk assessment of human Toxoplasmosis caused by ingesting raw or improperly cooked pork and pork products is not available. The aim of this study was to show current data on T. gondii prevalence in German pig farms. In four pig farms with different management systems (three conventional, one organic) 100 animals each were selected and tested for T. gondii antibodies. The test was done four times during the period from birth to slaughtering. In one farm 20 mother sows were tested additionally. The slaughtered pigs from conventional farms showed seroprevalences between 0 and 15.2% (mean value 5.6%). At the organic system T. gondii antibodies were not detected. All slaughtered seropositive pigs (6 months old) were tested negatively at the age of 9 weeks, but shortly after birth high titres of T. gondii antibodies had been detected in the same animals. Comparing the results gained in different seasons significantly more pigs were found to be infected during the autumn/winter than in the spring/summer period. In order to assess the current risk of Toxoplasmosis more pig farms should be tested. From the point of view of consumer protection the detection of highly infected pig herds is necessary.     

Infection dynamic of Toxoplasma gondii in two fattening pig farms exposed to high and low cat density in an endemic region

The presence of cats in the farms is considered a risk factor for the infection of pigs with Toxoplasma gondii (T. gondii). Cats eliminate oocysts that contaminate food, water and promote the infection of host reservoir such as rodents and birds among others that are also involved in the infection of pigs. The objective of this study was to assess the dynamic of infection of T. gondii in seronegative weaned pigs from weaning to 20 weeks of age from two farms from an endemic region, one with high and low density of cats. A cohort study was performed in 64 pigs, 31 newly weaned pigs on a farm with a high density of cats (FA) and 33 newly-weaned pigs on a farm with a low density of cats (FB). Blood samples were collected every 14 days to determine the presence of IgG antibodies against T. gondii in the serum using an indirect ELISA test. True incidence rate (TIV), cumulative incidence (AI) and relative risk (RR) was calculated. The age of seroconversion was determined by using survival tables; both farms were compared with Long-Rank test. In FA 97.5% of the pigs seroconverted at the second sampling and 100% at the third sampling, while in the FB all pigs seroconverted to the fourth sampling. The TIV was 0.67 and 0.43 for FA and FB respectively, during the first four weeks at risk. A RR of 1.5 (1.04-2.39) was obtained (p<0.05). Animals of the FA had a higher risk of infection compared with the FB, however, all animals included in the study had contact with the agent. Infection with T. gondii was rapidly distributed in both farms, regardless of the relative density of cats observed during the study. These results suggest a high environmental contamination with oocysts in the facilities of both farms probably due to the fact that T. gondii infection is endemic in the area where the farms are located, allow proper establishment of the etiological agent. The points of prevention and control strategies to avoid exposure of pigs to T. gondii in an endemic area should focus on the control of cats and rodents.     

Correlation of tissue infection and serologic findings in pigs fed Toxoplasma gondii oocysts

Each of thirteen 6-week-old pigs was inoculated per os with 10,000 sporulated oocysts of Toxoplasma gondii. By postinoculation day (PID) 13, pigs were seropositive by the indirect fluorescent antibody test. Beginning on PID 13 and every 7 days thereafter through PID 97, 1 pig was killed and 6 tissues were examined for T gondii. Of the 13 pigs, 11 were infected, including the 1st pig killed on PID 13, although none of the pigs had gross lesions of toxoplasmosis. Tissues harboring T gondii most frequently were the heart and brain; organisms were detected less frequently in the longissimus muscles, diaphragm, and liver. Toxoplasma gondii was not detected in the bronchial lymph nodes. There was good correlation between antibody and presence of T gondii in these pigs. One additional pig, maintained as a noninfected control, remained seronegative and had no evidence of infection when killed on PID 97.     

Diagnosis of transplacentally induced toxoplasmosis in pigs

Seventeen sows were fed 1,000 Toxoplasma gondii oocysts of isolates GT-1 or PT-1 at 32 to 92 days of gestation, and the products of conception were examined for T gondii antibodies and parasites. Twelve of these sows were euthanatized near term between 21 and 62 days after being fed T gondii; fetal body fluids or fetal sera were examined for agglutinating T gondii antibodies, and tissues were bioassayed in mice for T gondii parasites. Six sows produced pigs that had been transplacentally infected with T gondii; one of them aborted a T gondii-infected fetus 17 days after ingesting oocytes. Agglutinating antibodies were detected in fetuses infected in utero, but transplacental transfer of T gondii antibodies was not observed in noninfected fetuses. Transcolostrally acquired T gondii antibodies disappeared by 3 months of age. Diagnosis of transplacental toxoplasmosis was confirmed on the basis of detection of T gondii organisms in fetal tissues by use of histologic examination and bioassay in mice. In conclusion, finding of T gondii antibodies in body fluids could serve as a rapid screening test for transplacental T gondii infection in pigs.     

IFN-γ expression and infectivity of Toxoplasma infected tissues are associated with an antibody response against GRA7 in experimentally infected pigs

Toxoplasma gondii, an obligate intracellular parasite, can be transmitted to humans via the consumption of infected meat. However, there are currently no veterinary diagnostic tests available for the screening of animals at slaughter. In the current work, we investigated whether cytokine responses in the blood, and antibody responses against recombinant T. gondii GRA1, GRA7, MIC3 proteins and a chimeric antigen EC2 encoding MIC2-MIC3-SAG1, are associated with the infectivity of porcine tissues after experimental infection with T. gondii. Two weeks after experimental infection of conventional 5-week-old seronegative pigs, an IFN-γ response was detected in the blood, with a kinetic profile that followed the magnitude of the GRA7 antibody response. Antibody responses to GRA1, MIC3 and EC2 were very weak or absent up to 6 weeks post infection. Antibodies against GRA7 occurred in all infected animals and were associated with the presence of the parasite in tissues at euthanasia a few months later, as demonstrated by quantitative real-time PCR and isolation by bio-assay. Remarkably, although brain and heart tissue remained infectious, musculus gastrocnemius and musculus longissimus dorsi were found clear of infectious parasites 6 months after experimental infection. Seropositive response in a GRA7 ELISA indicates a Toxoplasma infection in pigs and is predictive of the presence of infectious cysts in pig heart and brain. This new ELISA is a promising tool to study the prevalence of Toxoplasma infection in pigs. Clearance of the infection in certain pig tissues suggests that the risk assessment of pig meat for human health needs further evaluation.     

The role of rodents and shrews in the transmission of Toxoplasma gondii to pigs

Inadequate rodent control is considered to play a role in Toxoplasma gondii infection of pigs. This issue was addressed in the current study by combining a 4-month rodent control campaign and a 7-month longitudinal analysis of T. gondii seroprevalence in slaughter pigs. Three organic pig farms with known rodent infestation were included in the study. On these farms, presence of T. gondii in trapped rodents was evaluated by real-time PCR. All rodent species and shrews investigated had T. gondii DNA in brain or heart tissue. Prevalence was 10.3% in Rattus norvegicus, 6.5% in Mus musculus, 14.3% in Apodemus sylvaticus and 13.6% in Crocidura russula. Initial T. gondii seroprevalence in the slaughter pigs ranged between 8% and 17% and dropped on the three farms during the rodent control campaign to 0-10%, respectively. After 4 months of rodent control, T. gondii infection was absent from pigs from two of the three farms investigated and appeared again in one of those two farms after the rodent control campaign had stopped. This study emphasizes the role of rodents and shrews in the transmission of T. gondii to pigs and the importance of rodent control towards production of T. gondii-free pig meat.