Cross-sectional survey on Toxoplasma gondii infection in cattle, sheep and pigs in Serbia: seroprevalence and risk factors

Toxoplasmosis is a globally distributed zoonosis with a clinical impact in the unborn fetus and in the immunosuppressed individual. In Serbia, studies of risk factors for Toxoplasma gondii infection in humans have shown that the relatively high prevalence is associated mainly with consumption of undercooked meat and/or meat products. However, data on T. gondii infection in domestic animals mostly used for human consumption are scarce. We thus conducted a cross-sectional survey on the seroprevalence of T. gondii infection in a representative sample of cattle, sheep and pigs from different regions of Serbia between June 2002 and June 2003, and analyzed the main risk factors associated with the infection. Sera from 611 cattle (yearlings and adults of both sexes), 511 ewes, and 605 pigs (market-weight and sows), were examined for T. gondii antibodies by the modified agglutination test. The seroprevalences determined were 76.3% in cattle, 84.5% in sheep and 28.9% in pigs. The antibody levels ranged from 1:25 to 1:400 in cattle, and up to 1:25,600 in sheep and to 1:12,800 in pigs. Among the seropositive, the proportion of high antibody levels (> or =1:1600), suggestive of acute infection, was 10% in sheep, and 4% in pigs. Possible association of the infection with biologically plausible risk factors including gender, age, herd size/farm type, type of housing, feeding practices and region, was analyzed by univariate analysis, and variables significant at P< or =0.1 were included in multivariate logistic regression models. The results showed that risk factors for cattle were small herd size (odds ratio, OR=2.19, 95% confidence interval, CI=1.28-3.75, P=0.004) and farm location in Western Serbia (OR=2.04, 95% CI=1.10-3.79, P=0.024), while housing in stables with access to outside pens was protective (OR=0.37, 95% CI=0.21-0.67, P=0.001). In sheep, an increased risk of infection was found in ewes from state-owned flocks (OR=4.18, 95% CI=2.18-8.00, P<0.001) vs. private flocks, and, interestingly, also in those from Western Serbia (OR=4.66, 95% CI=1.18-18.32, P=0.028). In pigs, the risk of infection was highly increased in adult animals (OR=3.87, 95% CI=2.6-5.76, P<0.001), as well as in those from finishing type farms (OR=3.96, 95% CI=1.97-7.94, P<0.001). In addition to providing data on the current T. gondii seroprevalence in meat animals in Serbia, the results of this study show the main risk factors associated with infection, thereby pointing to the type of preventive measures to reduce T. gondii infection.     

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.     

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.     

Seroprevalence of Toxoplasma gondii antibodies in free-ranging mithuns (Bos frontalis) from India

Studies conducted on free-ranging mithuns of Nagaland, India revealed that the overall seroprevalence of antibodies to Toxoplasma gondii in mithun was 42% (95% CI = 33-51) when detected by modified direct agglutination test. Highest (prevalence = 57%, 95% CI = 43-71) seroprevalence was found in mithuns above 3 years of age and the lowest (prevalence = 18%, 95% CI =4-32) in mithuns of 1-2 years old. No statistically significant difference was observed between male (prevalence = 40%, 95% CI = 26-54) and female (prevalence = 44%, 95% CI = 31-57) seroprevalences. The study also revealed that the maximum number (57%) of animals with the highest titre (1 : 3200) were above 3 years of age. This is the first serological survey for Toxoplasma gondii antibodies in free-ranging mithuns from India.     

Seroprevalence of Toxoplasma gondii in wild pigs (Sus scrofa) from Spain

Sera collected from 507 hunter-killed wild pigs (Sus scrofa) between 1993 and 2004 from five geographic regions in northern Spain and seven regions in southern Spain were assayed for antibodies to Toxoplasma gondii by the modified agglutination test (MAT). Antibodies to T. gondii were detected in 185 (38.4%) of 507 pigs with titers of 1:25 in 71, 1:50 in 111 and > or =1:500 in 3; seroprevalence was significantly higher (P<0.05) in pigs from southern regions. Seroprevalence was density dependent; it was higher in pigs from high stocking per hectare and availability of forage. Statistically significant differences were not observed between T. gondii seroprevalence and hunting estates (open versus fenced), sex or age. Serological results indicate a widespread exposure to T. gondii among Spanish wild boars, suggesting that this population could represent a public health risk for persons that handle or consume raw or undercooked infected wild pig meat.     

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。     

Prevalence of Toxoplasma gondii antibodies in southern Louisiana swine in 1980 and 1981

Sera from 1,219 pigs slaughtered in southern Louisiana in 1980 and 1981 were tested, using the indirect hemagglutination test and microtitration system, for Toxoplasma gondii antibodies; 19.19% had titers of greater than or equal to 1:64. Pigs slaughtered in the colder months, November to March, had significantly higher seroprevalences than did pigs slaughtered in the warmer months. Examination of blood samples from 236 sows and gilts on 52 farms revealed a seroprevalence of 24.6%.     

Prevalence of Toxoplasma gondii antibodies in wild red deer, roe deer, moose, and reindeer from Norway

Serum samples from 4339 wild cervids collected in Norway were tested for antibodies against Toxoplasma gondii using the direct agglutination test. The association between seroprevalence and species, sex, age, and geographic region was investigated. Positive titers (> or =1:40) were found in 33.9% of 760 roe deer (Capreolus capreolus); 12.6% of 2142 moose (Alces alces); 7.7% of 571 red deer (Cervus elaphus); and 1.0% of 866 reindeer (Rangifer tarandus). The seroprevalences were significantly different between the species. There was a significant increase in prevalence with age in roe deer, moose, and red deer, except from yearling to adult in red deer. A significant age-sex interaction was found in moose, and the effect of age was most distinct for females. No association between seropositive animals and sex was found for roe deer and red deer. There were significant differences in prevalence between geographic regions in roe deer and male moose. A widespread exposure to T. gondii in Norwegian cervids is documented, and meat from Norwegian cervids, particularly roe deer, should be regarded a potential source of infection for humans.     

Survey to determine the seroprevalence of Toxoplasma gondii infection in British sheep flocks

A serological survey of Toxoplasma gondii infection in adult breeding sheep in Great Britain was conducted using surplus sera taken during a seroprevalence study of Brucella melitensis in 2009. Of the 3539 sera collected from 227 flocks, 2619 (74 per cent) were found to be positive for T gondii specific antibody when tested using latex agglutination. Multilevel logistic modelling suggested that the likelihood of infection increased with age and this effect appeared to be amplified in animals vaccinated against T gondii. The model also indicated that the odds of sheep being seropositive were increased on premises where cattle were also kept. These results suggest a high level of Toxoplasma infection in breeding sheep in Great Britain and provide further evidence to suggest that postnatal infection is more common than congenital infection in sheep.     

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.     

Seroepidemiology of leptospirosis, toxoplasmosis, and leishmaniosis among dogs in Ankara, Turkey

Seroprevalence of five different Leptospira interrogans serovars, Toxoplasma gondii and Leishmania infantum in stray dogs in Ankara was investigated. A total of 116 dog sera collected from apparently healthy stray dogs were tested for L. interrogans serovars by microscopic agglutination test (MAT), for T. gondii antibodies by Sabin-Feldman dye test (SFDT), and for L. infantum antibodies by indirect fluorescence antibody test (IFAT). Of the 116 dogs, 51 (43.96%) were seropositive for leptospirosis, 72 (62.06%) for T. gondii and 3 (2.58%) for L. infantum. No statistically significant difference was observed between male and female dogs in the seroprevalences of toxoplasmosis and leptospirosis (P>0.05), but statistically significant difference was observed among different age groups in the seroprevalences of toxoplasmosis and leptospirosis (P<0.05). Although the seroprevalence of L. infantum was low, asymptomatic animals should be considered as a reservoir for the spread of the disease.     

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.     

The prevalence of serum antibodies to Toxoplasma gondii in Ontario mammals.

The prevalence of seropositive reactions to Toxoplasma gondii was studied in farm animals, companion animals, wild rodents and birds. Of the animals tested, 17% of cattle, 65% of sheep, 45% of pigs, 9% of horses, 33% of dogs and 20% of cats were seropositive by the Sabin-Feldman dye test. In addition 11% of mice (Mus musculus), 5% of deer mice (Peromyscus), 3% of rats (Rattus norvegicus) and less than 2% of sparrows (Passer domestcus) were seropositive. All samples from short-tailed field mice (Microtus pennsylvanicus), squirrels (Sciurus carolinensis), chipmunks (Tamias striatus), meadow jumping mice (Zapus hudsonius) and starlings (Sturnus vulgaris) were seronegative. The significance of these findings in relation to the epizootiology of toxoplasmosis in Ontario is discussed.     

Seroprevalence of Hepatitis E Virus in Domestic Pigs and Wild Boars in Switzerland

Hepatitis E is considered an emerging human viral disease in industrialized countries. Studies from Switzerland report a human seroprevalence of hepatitis E virus (HEV) of 2.6–21%, a range lower than in adjacent European countries. The aim of this study was to determine whether HEV seroprevalence in domestic pigs and wild boars is also lower in Switzerland and whether it is increasing and thus indicating that this zoonotic viral infection is emerging. Serum samples collected from 2,001 pigs in 2006 and 2011 and from 303 wild boars from 2008 to 2012 were analysed by ELISA for the presence of HEV‐specific antibodies. Overall HEV seroprevalence was 58.1% in domestic pigs and 12.5% in wild boars. Prevalence in domestic pigs was significantly higher in 2006 than in 2011. In conclusion, HEV seroprevalence in domestic pigs and wild boars in Switzerland is comparable with the seroprevalence in other countries and not increasing. Therefore, prevalence of HEV in humans must be related to other factors than prevalence in pigs or wild boars.     

Toxoplasma gondii seroprevalence in domestic animals and humans in Mymensingh District, Bangladesh

The seroprevalence of Toxoplasma gondii was examined in adult women and domestic animals used for meat products from the Mymensingh District, Bangladesh. Cattle, goats and sheep showed a high seroprevalence (12, 32 and 40%, respectively), while the sera from all fifteen women examined in the same area were seronegative. Considering that primary infection in women during pregnancy can cause abortion and congenital defects, accidental ingestion of T. gondii infected meat products from domestic animals represents a risk factor for adult women living in the same area.     

Prevalence of antibodies to toxoplasma gondii in cattle and swine in the Netherlands: Towards an integrated control of livestock production

Summary

Serological surveys of the prevalence of antibodies against Toxoplasma gondii were carried out amongst swine and cattle in the Netherlands. Data were analysed according to the different categories of animals. The results show very low seroprevalences of Toxoplasma gondii in finishing pigs (1.8%) and in fattening calves (1.2%). In sows and dairy cattle, respectively, seroprevalences of 30.9% and 27.9% respectively, were found, demonstrating clearly the environmental infection pressure and illustrating the importance of housing and management in establishing low infection rates.

Substantially different seroprevalences were found between dairy cattle sampled in the North and in the South of the Netherlands (13.1% and 42.6%, respectively).

The infection rates in the samples from finishing pigs, fattening calves, and dairy cattle demonstrate that seroprevalences in individual farms or herds may differ considerably. Investigation of the factors involved can be useful in determining the causes of infection and for developing measures with regard to prevention. The very low seroprevalences in finishing pigs and fattening calves indicate, however, that the production of toxoplasma‐free meat may be well within reach in modern husbandry.

Since farm animals easily are infected, serological screening of individual farms or herds for the absence of T. gondii infection, as a part of the Integrated Quality Control programme, can be helpful in determining the quality of livestock production and in developing certain standards of hygiene for individual farms.     

Toxoplasma gondii infection in sheep from Haute-Vienne, France: seroprevalence and isolate genotyping by microsatellite analysis

Ingesting meat of free-range livestock, mainly sheep, is associated with human toxoplasmosis in European countries. Data on Toxoplasma gondii infection in French ovine livestock are relatively scarce. Sera from 164 lambs and 93 ewes slaughtered in Haute-Vienne district, France, were tested by a direct agglutination test. Antibodies to T. gondii were found in 36 (22.0%) lambs and in 61 (65.6%) ewes. In addition, to attempt parasite isolation for genotyping, hearts from 50 other ewes were obtained from a local slaughterhouse, and were screened by a direct agglutination test. T. gondii was isolated in 8 of 30 seropositive hearts bioassayed in mice. All isolates were type II by genetic characterization at five microsatellite loci (TUB2, TgM-A, W35, B17, B18). These results indicate that bovines slaughtered in France may be highly infected by T. gondii with a potential risk of parasite transmission to humans by consumption of undercooked meat. Multilocus microsatellite analysis shows the predominance of type II in sheep as previously described in humans.     

内蒙古部分地区放牧牛羊弓形虫血清学调查

[目的]了解内蒙古部分地区放牧牛羊弓形虫病感染情况。[方法]采用间接血凝试验(IHA)对阿拉善盟及呼伦贝尔市随机采集的286份放牧牛羊血清样本进行弓形虫抗体检测。[结果]绵羊血清弓形虫抗体总阳性率为1.68%,其中,阿拉善盟阳性率为4.84%,呼伦贝尔市阳性率为0.56%;经统计学分析,两个地区的绵羊血清弓形虫抗体阳性率差异不显著(P>0.05);牛血清弓形虫抗体总阳性率为0。[结论]内蒙古主要牧区放牧绵羊存在弓形虫感染,应引起足够的重视,并制定相应的弓形虫病防治措施。     

The influence of feeding and maintenance system on occurrence of Toxoplasma gondii infections in dogs

Serum samples of 113 dogs visiting "outpatient clinics", 52 dogs kept in shelters and 35 animals from a military dog training centre were examined for Toxoplasma gondii specific antibodies using a latex agglutination test. Significant differences in seroprevalences were found between dogs from the training centre (8.6% of positive results) and the other populations examined (40.7% of positive seroreagents in animals visiting outpatient clinics and 44.2% in the group from shelters, respectively). Among clinic patients, dogs fed raw meat were significantly more frequently seropositive (65.2%) than those eating only commercial dry feed or cooked meat (25.7%). No statistically significant differences were noted in males compared to females and in pure breed dogs compared to crossbreed dogs. The antibodies were usually found in low titres under 60 IU/ml (69.6% of positive results). High titres (120-480 IU/ml) were detected in 2 of 3 dogs with clinical toxoplasmosis. In these dogs IFAT T. gondii specific IgM were detected and a favourable response to antiprotozoal treatment was observed. All the dogs with medium and high titres were given raw meat. Age and the presence of cats did not seem to have any influence on T. gondii seroprevalence. Neospora caninum specific antibodies in low titres ranging from 1:20-1:320 were found in 7 (9.7%) of 72 T. gondii positive seroreagents.     

Seroprevalence of Toxoplasma gondii infection in domestic goats in Satun Province, Thailand

Goats are important domestic animals in the south of Thailand due to the minimal cost of rearing and maintaining them, and their production of both meat and milk. Toxoplasmosis is one of the most threatening parasitic zoonoses and the causative agent Toxoplasma gondii uses a wide range of warm-blooded intermediate hosts including the goat. The objective of this study was to assess the seroprevalence of antibodies to T. gondii in goats of Satun Province in Thailand. A total of 631 goat sera were examined for antibodies against toxoplasmosis with commercial latex agglutination test kits (Toxocheck-MT 'Eiken'). Of these, 176 (27.9%) were found to be positive to T. gondii; antibody titers ranged from 1:64 to 1:4096 (1:64 cut-off). Female goats were 1.73 times more likely than male to be seropositive (odds ratio [OR]=1.73; 95% confidential interval [CI]=1.11, 2.73). Dairy goats were more likely to be seropositive than meat goats (OR=1.36; 95% CI=0.84, 2.20). Goats were infected with T. gondii with acquisition of age because older goats were more likely to be seropositive than young goats under 1-year-old (for 1-2 years, OR=19.6; 95% CI=0.92, 4.15, for >2 years, OR=2.70; 95% CI=1.26, 5.80). The high seroprevalence of T. gondii antibodies found in the present study suggested widespread exposure of goats in Satun Province to T. gondii.