Modified Agglutination Test Research Articles

Comparison of Seroprevalence and PCR Results in the Detection of Toxoplasma gondii in Pet Rabbits in Poland.

Toxoplasma gondii infections are prevalent in humans and animals worldwide. The aim of the study was to estimate the seroprevalence of T. gondii in pet rabbits, as well the presence of T. gondii DNA in their blood. A total of 360 pet rabbits were investigated for the presence of antibodies and antigens of T. gondii in blood samples using a modified agglutination test (MAT) and nested PCR, respectively. Antibodies against T. gondii were found in 44 (12.12%) of pet rabbits. In rabbits that received unwashed vegetables 13.13% were positive for T. gondii antibodies, whereas all rabbits that received washed vegetables were seronegative. The prevalence of anti-T. gondii antibodies was statistically higher in samples collected from ill rabbits (45.45%) compared with healthy ones (8.87%), as well in the group of rabbits that had contact with cats (16.13%) compared with those without contact with cats (3.57%). The percentage of seropositive samples derived from all three sampling regions of Poland was as follows: Silesia (12.20%), Lower Silesia (6.09%), and Lesser Poland (18.03%). No statistical differences in seroprevalence were observed according to age or sex of rabbits. All tested blood samples were negative for the T. gondii B1 gene in nested PCR. This is the first study of seroprevalence and the presence of T. gondii in the blood of pet rabbits in Poland. Our study indicates that health status of rabbits, contact with cats, as well sampling region may have an important impact on the prevalence of T. gondii infection.

SURVEI SEROLOGIS TOXOPLASMOSIS PADA TERNAK SAPI DI KABUPATEN PRINGSEWU MENGGUNAKAN METODE TOXOPLASMA MODIFIED AGGLUTINATION TEST (To-MAT)

The cattle population in Lampung Province has a relatively high number when compared to several other provinces. Pringsewu is one of the districts in Lampung Province having the supplier potential of beef in Lampung. The higher the population and production, could cause a potential infection of toxoplasmosis to livestock and human. So far the prevalence of toxoplasmosis in Pringsewu District. This study was conducted to determine the level of toxoplasmosis infection in cattle in Pringsewu district. This study used the To-MAT (Toxoplasma Modified Agglutination Test) method to test Toxoplasma gondii infection in blood of cows samples. The result of the study found that at a chronic level there were 23/24 cows positively infected with T.gondii. Whereas in acute level there are 8/24 cows positively infected with T. gondii. In general, the prevalence of toxoplasmosis is 95.8%. Keywords : Toxoplasmosis, Cow, To-MAT (Toxoplasma Modified Agglutination Test) method, Infection

Serological evidence of Toxoplasma gondii and Neospora caninum infection in black-boned sheep and goats in southwest China

Toxoplasma gondii and Neospora caninum are two closely related protozoan parasites which can cause abortion and significant economic losses in sheep and goats. However, it is yet to know whether black-bone sheep and goats are infected with T. gondii and N. caninum in China. In the present investigation, the seroprevalence and risk factors of T. gondii and N. caninum infections in black-boned sheep and goats were investigated in Yunnan Province, subtropical southwest China between July and August of 2017. A total of 481 serum samples were tested for T. gondii antibodies using the Modified Agglutination Test (MAT), and 468 serum samples were examined for N. caninum antibodies by indirect Enzyme-Linked Immunosorbent Assay (iELISA). The overall seroprevalence of T. gondii in black-boned sheep and goats was 36.80% (177/481, 95% CI 32.49–41.11), and 40 out of 468 serum samples were N. caninum-seropositive (8.55%, 95% CI 6.02–11.08). There was significant difference in the seroprevalence of T. gondii infection in different regions (χ2 = 19.869, df = 2, P<0.01). As for the seroprevalence of N. caninum infection, region (χ2 = 8.558, df = 2, P<0.05), age (χ2 = 16.631, df = 3, P < 0.01), gender (χ2 = 11.219, df = 1, P < 0.01) and species (χ2 = 8.673, df = 1, P < 0.01) were the risk factors. In addition, the seroprevalence of coinfection of T. gondii and N. caninum in black-boned sheep and goats was 3.63% (17/468, 95% CI 1.94–5.32). To our knowledge, this is the first report of T. gondii and N. caninum seroprevalence in black-boned sheep and goats in China, which provided base-line data for the execution of control strategies and measures against T. gondii and N. caninum infection in black-boned sheep and goats.

Genotyping of viable Toxoplasma gondii from the first national survey of feral swine revealed evidence for sylvatic transmission cycle, and presence of highly virulent parasite genotypes.

Feral swine are known reservoirs of various pathogens, including Toxoplasma gondii. Here, we report the first national survey of viable T. gondii in feral swine in the USA. We paired serological surveys with parasite isolation and bioassay to evaluate the prevalence and genetic diversity of these parasites. From 2012-2017, sera and tissues from 1517 feral swine across the USA were collected for the isolation of viable T. gondii. Serum samples were initially screened for antibodies to T. gondii, and then the tissues of seropositive feral swine were bioassayed in mice. Antibodies were detected in 27.7% of feral swine tested by the modified agglutination test (1:25 or higher). Antibody positive rates increased significantly with age, with 10.1% of juveniles, 16.0% of sub-adults and 38.4% of adults testing seropositive. Myocardium (50 g) from 232 seropositive feral swine was digested in pepsin and bioassayed in mice. Viable T. gondii was isolated from 78 feral swine from 21 states. Twelve of the 78 isolates were pathogenic to outbred Swiss Webster mice and 76 of the 78 isolates could be propagated further in cell culture and were genotyped. For genotyping, deoxyribonucleic acid extracted from cell culture-derived tachyzoites was characterized by polymerase chain reaction restriction fragment length polymorphism using the genetic markers SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genotyping revealed 15 ToxoDB genotypes, including 43 isolates for genotype #5 (haplogroup 12), 11 isolates for #24, four isolates for #2 (haplogroup 3), two isolates for each of genotypes #3 (haplogroup 2), #4 (haplogroup 12), #216, #221, #289 and #297 and one isolate for each of genotypes #1 (haplogroup 2), #39, #66, #260, #261 and #299. Genotype #5 was the most frequently isolated, accounted for 57% (43/76) of the isolates, followed by #24, accounted for 14% (11/76). Genotypes #260, #289, #297 and #299 are new types. Genotype #289 was highly virulent to mice and originated from feral swine collected in Louisiana on the same day at the same location. Genotype #216 was previously demonstrated to be highly virulent to mice. Our results indicate moderate genetic diversity of T. gondii in feral swine in the USA, with the genotype #5 (haplogroup 12) dominant in the continental USA, whereas genotype #24 (10/14) was dominant in Hawaii, suggesting different population structures of the parasites among the two distinct geographical locations.

High Seroprevalence But Low Rate of Isolation of Toxoplasma gondii from Wild Elk (Cervus canadensis) in Pennsylvania

Toxoplasma gondii infections are prevalent in most warm-blooded animals worldwide. During the 2018 November hunting season in Pennsylvania, fresh (unfixed, not frozen) samples obtained from 99 harvested elk (Cervus canadensis) were tested for T. gondii infection. Antibodies to T. gondii were detected in 69 of 99 (69.7%) elk tested by the modified agglutination test (MAT, 1:25 cut-off). Tongues and hearts from 16 elk with high MAT titers (>1:200) were bioassayed for T. gondii by inoculation in outbred Swiss Webster (SW) and interferon-gamma gene knockout (KO) mice. Viable T. gondii was isolated from tongues of 2 elk with MAT titers of 1:200 and 1:3,200. Toxoplasma gondii from both isolates were successfully propagated in cell culture. Genetic typing on DNA extracted from culture-derived tachyzoites using the PCR restriction fragment length polymorphism with 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) revealed that both isolates belonged to ToxoDB PCR-RFLP genotype #5 that is widely prevalent in wildlife in the United States. Our results suggest that elk may clear T. gondii organisms from their tissues.

Seroprevalence of anti-Toxoplasma gondii antibodies in wild boars (Sus scrofa), hunting dogs, and hunters of Brazil

Seroprevalence of Toxoplasma gondii has been extensively studied in wild boars worldwide due to the emerging risk for human infection through meat consumption. However, this is the first study that reports toxoplasmosis seroprevalence in wild boars, wild boar hunters and their hunting dogs. The aim of the present study was to evaluate the seroprevalence of anti-T. gondii antibodies in the complex wild boars, hunting dogs and hunters, and to determine the risk factors associated with seropositivity in southern and central-western Brazil. Overall, anti-T. gondii seropositivity was observed in 15/71 (21.1%) wild boars by modified agglutination test (MAT); and 49/157 (31.2%) hunting dogs and 15/49 (32.7%) hunters by indirect immunofluorescent antibody test (IFAT). Seroprevalence of toxoplasmosis in Brazilian wild boars was within the national and international range, posting wild boars as potential environmental sentinels for T. gondii presence. In addition, the findings have comparatively shown that wild boars have been less exposed to infection than hunting dogs or hunters in both Brazilian regions. Seropositivity for T. gondii was statistically higher in 12/14 (85.7%) captured wild boars when compared to 5/57 (7.0%) free-range wild boars (p = 0.000001). Similarly, captured wild boars from anthropized areas were more likely to be seropositive than of natural regions (p = 0.000255). When in multiple regression model, dogs with the habit of wild boar hunting had significant more chance to be positive (adjusted-OR 4.62 CI 95% 1.16-18.42). Despite potential as sentinels of environmental toxoplasmosis, seroprevalence in wild boars alone may provide a biased basis for public health concerns; thus, hunters and hunting dogs should be always be included in such studies. Although hunters should be aware of potential T. gondii infection, wild boars from natural and agricultural areas may present lower protozoa load when compared to wild boars from anthropized areas, likely by the higher presence of domestic cats as definitive hosts.

Mat Seroprevalence Infers Low Rates of Toxoplasma gondii in Domestic Pigs from Yucatan, Mexico

Toxoplasma gondii is a zoonotic parasite of worldwide distribution. The consumption of infected pork meat has been suggested to be an important source for human infection in the tropical area of Yucatan, Mexico. We performed a cross-sectional study of 12 farms across the state to investigate the seroprevalence of Toxoplasma gondii infection in domestic pigs. In total, 632 samples were obtained from 2 different environmental zones (tropical deciduous low forest and tropical sub-deciduous medium forest) and 2 abattoirs. The modified agglutination test (MAT) was used to assess the seroprevalence of T. gondii in pigs and to evaluate 2 globally used serological tests, the Dye test (DT) and ID Screen® ELISA multi-species, and a commercial ELISA kit (Human Toxo IgG, Human-diagnostics), which is widely used locally in this geographical area. The overall prevalence obtained with the MAT (cut-off ≥1:25) among the 632 pigs was 1.4% (95% CI, 0.6-2.7%). The seroprevalence obtained for the different age groups was 0.6%, 0.7%, 1.8%, and 6.8% among 2-3, 3-4, 4-5, and ≥5-mo-old pigs. This increase in the seroprevalence was statistically significant for the 2 older groups (odds ratio [OR] 3.9-7.1, P < 0.05) in comparison with younger groups. DT at >4 IU dilution had a perfect agreement and 100% of sensitivity and specificity when compared with the MAT. Although ID Screen® had only a fair agreement (κ = 0.389) with the MAT, the McNemar test showed that the results of these tests were comparable (P = 0.29). The Human Toxo ELISA showed no agreement with MAT, ID Screen®, and DT (κ = 0.000-0.023, McNemar P < 0.05). This ELISA was lacking in specificity, accuracy, and precision; hence, we do not recommend its use for T. gondii diagnosis in pig serum.

Prevalence of Anti–Toxoplasma gondii Antibodies in Ratites from Brazil

The prevalence of anti-Toxoplasma gondii antibodies was investigated among 104 ratites: 68 rheas (Rhea americana), 16 emus (Dromaius novaehollandiae), and 20 ostriches (Struthio camelus) in 4 Brazilian states. The prevalence in rheas was 26.5% (18 of 68), and titers were 1:100 (n = 8), 1:200 (n = 1), 1:400 (n = 4), 1:800 (n = 4), and 1:1,600 (n = 1). In emus, the prevalence was 50% (8 of 16), and titers were 1:50 (n = 1) and 1:100 (n = 7). The ostriches were slaughtered for human consumption, and 80% (16 of 20) were seropositive with titers of 1:200 (n = 1), 1:400 (n = 9), and 1:800 (n = 6). Sera were tested with a modified agglutination test, and the results confirmed the distribution of the parasite in ratite species from Brazil. The data obtained in this study show that T. gondii is prevalent among ratites from Brazil, and therefore ratite meat should also be considered a potential source of human infection. This is the first report of T. gondii seroprevalence in emus.

First report on seroprevalence and risk factors of Toxoplasma gondii infection in sheep and goats in North Lebanon.

Toxoplasmosis is of dual importance in both public and veterinary health due to the respective risk of transplacental transmission in primo-infected pregnant women and economic losses caused by abortions in mammals. One of the main routes of Toxoplasma gondii transmission to humans is the consumption of raw or undercooked meats containing parasitic cysts. Here, we performed the first epidemiological study to determine the seroprevalence and the risk factors of toxoplasmosis in livestock in Lebanon. Using a modified agglutination test with a cut-off of 1:40, we tested the positivity rate of Immunoglobulin G antibodies in the sera of 100 sheep and 80 goats collected from 18 different livestock farms located in North Lebanon between March and June 2018. Anti-Toxoplasma gondii IgG antibodies were detected in 42% of sheep and 34% of goats. Adults (> 1 year) were significantly more infected by T. gondii than the lambs (< 1 year) in both species (p < 0.05). These findings indicated that food animals are highly exposed to T. gondii in Lebanon and could be potentially a major risk factor of T. gondii infection to humans. Consequently, national prophylactic strategies should be implemented to control and to prevent T. gondii transmission between animals and humans.

Serological Studied On Toxoplasma Gondii Infection in Sheep and Goats in Ismailia Province

Blood serum samples from 100 sheep and 100 goats were collected and examined for Toxoplasma gondii antibodies by Enzyme Linked Immunosorbent Assay and Modified Agglutination Test. The seroprevalences of Toxoplasma gondii in sheep were 34% and 33% and in goats were 32%, 31% by ELISA and MAT respectively. The prevalence in the females of sheep and goats were higher than males. The seroprevalences were higher in adult animals than young in both sheep and goats. Using the MAT as a reference test, the sensitivity and specificity of the ELISA Test were 100% and 98.5% respectively.

Variation in Toxoplasma gondii seroprevalence: effects of site, sex, species and behaviour between insular and mainland macropods

Context Feral cats threaten wildlife conservation through a range of direct and indirect effects. However, most studies that have evaluated the impacts of feral cats on species of conservation significance have focussed on direct impacts such as predation; few studies have considered the indirect impacts of cat-borne disease. Toxoplasma gondii, a cat-borne parasite, causes both acute and latent disease in a range of wildlife species, and macropods are particularly susceptible. Kangaroo Island is Australia’s third largest island and supports a high density of feral cats and high seroprevalence of T. gondii in multiple species, relative to the mainland. This suggests that Kangaroo Island has a high environmental contamination with the parasite and a high risk of infection for other species. Aims We aimed to describe T. gondii seroprevalence in culled and road-killed macropods, so as to assess the effects of island versus mainland location, sex, species and behaviour. Methods Macropod sera were tested for T. gondii IgG antibodies using a commercially available modified agglutination test. Key results The seroprevalence of T. gondii in culled western grey kangaroos (Macropus fuliginosus) was significantly higher on the island (20%, 11/54 positive) than on the mainland (0%, 0/61 positive). There was no difference in T. gondii seroprevalence between culled and road-killed (21%, 21/102 positive) kangaroos from the island. The seroprevalence of T. gondii was significantly higher in female (32%, 12/38 positive) than in male (13%, 8/60 positive) kangaroos, but we observed no sex effect in tammar wallabies (Macropus eugenii), and no effect of species. Conclusions The higher T. gondii seroprevalence in insular macropods supports previous reports of higher T. gondii exposure in other Kangaroo Island fauna. The lack of difference in T. gondii seroprevalence between culled and road-killed kangaroos suggests that T. gondii-positive animals are not more vulnerable to road mortality, in contrast to that suggested previously. Implications Our findings suggest greater potential adverse conservation impacts owing to toxoplasmosis on the island than on the mainland. In light of a recent study demonstrating higher cat abundance on the island than on the mainland, the higher observed T. gondii seroprevalence in insular macropods is likely to be a consequence of higher cat density.

Toxoplasma gondii seroprevalence in endangered bridled nailtail wallabies and co-occurring species

The parasite Toxoplasma gondii can infect any warm-blooded species; however, seroprevalence in most species remains largely unknown. In this study we examined the presence of T. gondii antibodies in captured individuals in the two remaining wild populations and one captive population of endangered bridled nailtail wallabies (Onychogalea fraenata). Samples from cats (Felis catus), rabbits (Oryctolagus cuniculus) and dogs (Canis lupus) were also taken opportunistically during invasive species control at the two wild populations. Seventy-one wallabies, sixteen cats, four rabbits, and two dogs’ blood samples were tested for T. gondii using a modified agglutination test. Half of the tested feral cats (n = 8) were seropositive and all intermediate hosts were seronegative for T. gondii antibodies. This unexpected result suggests a loss of infected individuals before capture and testing, or parasite infectivity being suppressed by Queensland’s hot, dry climate.

Seroprevalence and Risk Factors of Toxoplasma gondii Infection in Free-Range Chickens in Senegal, West Africa.

In sub-Saharan Africa, few studies have addressed the environmental determinants of the incidence of Toxoplasma gondii infection. Free-range chickens are regarded as sensitive indicators for environmental contamination with T. gondii oocysts due to their ground-feeding behavior and have been used as sentinels. A cross-sectional study was conducted from January to April 2018 to estimate the seroprevalence of T. gondii infection in free-range chickens from Senegal, West Africa, using the modified agglutination test. Sampling was performed in two regions of the country: Saint-Louis, a Sahelian area in the North, and Kedougou, a forest and rainy area in the South. A questionnaire was administered to chicken owners to assess potential risk factors for T. gondii seropositivity, and univariable and multivariable logistic regression analyses were used to determine the statistical significance of risk factors. The seroprevalence in chickens was 7.67% (51/665; 95% confidence interval [CI]: 5.51-9.52). The multivariable logistic regression model indicated that the odds to test T. gondii seropositive was higher in chickens provided well water (odds ratio [OR] = 3.17, 95% CI: 1.45-6.93; p = 0.004) than in chickens provided tap water, and in hens having chicks (OR = 80.15, 95% CI: 22.79-281.95; p < 0.001) than in chickens (male or female) not having chicks. The possible role of contaminated well water in the acquisition of infection in chickens-and hence in human-merits consideration and should be addressed in future studies.

Prevalence of Toxoplasma gondii and Neospora caninum infections in households sheep "Elevage en case" in Dakar, Senegal.

Aim:The study aimed to evaluate the occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep breeding in a particular husbandry system called “Elevage en case” in Dakar, Senegal.Materials and Methods:Blood samples were collected from 278 sheep. Serum was harvested and used for analysis. For the detection of T. gondii antibodies, 278 sera were analyzed using the modified agglutination test, while the enzyme linked-immunosorbent assay was used on 174 sheep sera to look for N. caninum antibodies.Results:This study showed that 29.4±6.8% of sheep carried both T. gondii and N. caninum antibodies. The overall prevalence was 60.1±5.7% and 41.9±7.3% for toxoplasmosis and neosporosis, respectively. For toxoplasmosis, the prevalence was higher in Gueule Tapée (63.3%) than in Medina (58.9%), but the variation was not significant (p=0.45). Regarding the age of animals, the prevalence was significantly higher in animals over 2 years old compared to those under 2 years old (p=0.002). For neosporosis, the prevalence was significantly higher in Medina (48.67%) than Gueule Tapée (16.7%) (p=0.001), but non-significant variation was noted according to animal age.Conclusion:The study showed that sheep reared in households have carried antibodies of T. gondii and N. caninum. The prevalence was high and it means that consumption of meat from these animals is risky if the meat is eaten undercooked.

Antibodies to Toxoplasma gondii in slaughtered free-range and broiler chickens

The consumption of undercooked infected chicken can be a source of infection for humans and carnivores regarding the zoonotic protozoan parasite Toxoplasma gondii. Furthermore, free-range chickens are sentinels for the presence of T. gondii oocysts in the environment because they feed from the ground. By using the modified agglutination test (MAT), we investigated the presence of antibodies to T. gondii in 178 free-range and 170 broiler chickens raised indoors and slaughtered in Portugal. Prevalence of specific antibodies was 5.6% in free-range and 0.0% in broiler chickens raised indoors (p = 0.002).

Epidemiological Investigation of Toxoplasma gondii Infections in Commercial Sheep Flock in an Endemic Area for Ocular Toxoplasmosis in Southern Brazil.

Toxoplasma gondii is a widely distributed parasite and of great importance to human and animal health. The objective of this study was to assess the prevalence of T. gondii antibodies and risk factors associated with the infection in sheep in the Northwest region of the State of Rio Grande do Sul, Brazil; this region has a very high rate of human ocular toxoplasmosis. Ovine sera were tested by the modified agglutination test (cut-off 1:25). T. gondii antibodies were detected in 70.2% (224 of 319). According to the logistic regression, the most significant factors associated were age and cat access to food stock facility. Preventive measures are discussed to reduce the risk of transmission of this zoonosis.

Isolation, genotyping and pathogenicity of a Toxoplasma gondii strain isolated from a Serval (Leptailurus serval) in China.

Toxoplasma gondii typically causes lifelong chronic infection and has been identified in a variety of intermediate and definitive hosts. Felids are capable of serving as both intermediate and definite hosts for T.gondii infection. However, there is no direct evidence to prove that servals are the intermediate host of T.gondii. In this study, T.gondii antibodies were detected in a serval by a modified agglutination test (titer, 1:200). Viable T.gondii was isolated from the striated muscles of the serval. This strain was further propagated in cell culture and designated as TgServalCHn1. Genetic characterization of DNA derived from cell culture was performed by RFLP-PCR of 10 markers, as well as polymorphic ROP5 and ROP18 genes. Results showed that this strain of T.gondii belonged to the genotype ToxoDB#20. The ROP5 allele 4 and ROP18 allele 3 suggested that this strain was avirulent, which was further supported by infection in mice. Encephalitis, immune organ necrosis and focal mononuclear cell infiltration in multiple organs were the main pathology characteristics observed in BALB/C mice infected with the TgServalCHn1 strain. To our knowledge, the present study is the first to report the isolation of T.gondii from a serval, which gives direct evidence for servals serving as an intermediate host of T.gondii. The genotyping results revealed the presence of genotype ToxoDB#20 in central China, enriching the scope of the distribution of T.gondii genotypes in Asia.

Prevalence and Genetic Diversity of Toxoplasma gondii in Free-Ranging Chickens from the Caribbean

PurposeToxoplasma gondii is a zoonotic parasite capable of infecting a wide range of hosts. Free-range chickens are important sentinels in the epidemiology of this parasite as they feed from the ground and are likely to ingest oocysts shed in the faeces of infected cats. Atypical strains of T. gondii are known to dominate in South America where they are associated with more severe disease in humans, yet relatively little is known about the strains circulating in neighbouring Caribbean islands.MethodsIn this study, hearts and brains were collected from free-range chickens in Antigua and Barbuda (n = 45), Dominica (n = 76) and Trinidad (n = 41), and DNA was extracted for nested ITS1 PCR and PCR–RFLP. Sera were collected and screened for antibodies using the modified agglutination test (MAT).ResultsAntibodies to T. gondii were detected in 20.5, 38.2 and 17.1% of chickens in Antigua and Barbuda, Dominica and Trinidad, respectively. Toxoplasma gondii DNA was also detected by PCR in 24.4, 17.1 and 17.1% of chickens, respectively, giving an overall prevalence of 31.1, 42.1, and 29.3% for each of the 3 island nations. Results of PCR–RFLP revealed 2 new atypical genotypes (designated ToxoDB #281 and #282) and one Type III (ToxoDB #2) in chickens from Antigua. Partial genotyping of a further 8 isolates (7 from Antigua and one from Trinidad) revealed different allele-types at five or more markers for 7 of the isolates, suggesting atypical genotypes.ConclusionsThis is the first study to report the prevalence of T. gondii in free-range chickens in Antigua and Barbuda, Dominica and Trinidad and Tobago. It is also the first to report the presence of atypical genotypes in Antigua and Barbuda and Trinidad and Tobago.

The relationship between the presence of antibodies and direct detection of Toxoplasma gondii in slaughtered calves and cattle in four European countries

In cattle, antibodies to Toxoplasma gondii infection are frequently detected, but evidence for the presence of T. gondii tissue cysts in cattle is limited. To study the concordance between the presence of anti-T. gondii IgG and viable tissue cysts of T. gondii in cattle, serum, liver and diaphragm samples of 167 veal calves and 235 adult cattle were collected in Italy, the Netherlands, Romania and the United Kingdom. Serum samples were tested for anti-T. gondii IgG by the modified agglutination test and p30 immunoblot. Samples from liver were analyzed by mouse bioassay and PCR after trypsin digestion. In addition, all diaphragms of cattle that had tested T. gondii-positive (either in bioassay, by PCR on trypsin-digested liver or serologically by MAT) and a selection of diaphragms from cattle that had tested negative were analyzed by magnetic capture quantitative PCR (MC-PCR). Overall, 13 animals were considered positive by a direct detection method: seven out of 151 (4.6%) by MC-PCR and six out of 385 (1.6%) by bioassay, indicating the presence of viable parasites. As cattle that tested positive in the bioassay tested negative by MC-PCR and vice-versa, these results demonstrate a lack of concordance between the presence of viable parasites in liver and the detection of T. gondii DNA in diaphragm. In addition, the probability to detect T. gondii parasites or DNA in seropositive and seronegative cattle was comparable, demonstrating that serological testing by MAT or p30 immunoblot does not provide information about the presence of T. gondii parasites or DNA in cattle and therefore is not a reliable indicator of the risk for consumers.

Isolation and Genetic Characterization of Toxoplasma gondii from Tissues of Wild Turkeys (Meleagris gallopavo) in Pennsylvania

Toxoplasmosis in wild turkeys (Meleagris gallopavo) is of epidemiological interest because turkeys feed from the ground, and detection of infection in turkeys indicates contamination by oocysts in the environment. During the 2018 spring hunting season in Pennsylvania, fresh (unfixed, not frozen) samples were obtained from 20 harvested wild turkeys and tested for Toxoplasma gondii infection. Hearts from all wild turkeys and skeletal muscle from 1 were bioassayed for T. gondii by inoculation in outbred Swiss Webster (SW) and interferon-gamma gene knockout (KO) mice. Antibodies to T. gondii were detected in 1:5 dilution of neat serum from 5 of 15 wild turkeys and in fluid from the heart of 1 of 4 wild turkeys with the modified agglutination test (MAT); neat serum was not available from 4 wild turkeys. Viable T. gondii was isolated from hearts of 5 wild turkeys, 1 with MAT of 1:10, 1 with MAT of 1:5, and 3 seronegative (MAT < 1:5). Toxoplasma gondii was isolated from both heart and skeletal muscle in the 1 wild turkey that had skeletal muscle submitted. The KO mice inoculated with tissue from all 5 infected wild turkeys died or were euthanatized when ill, 7-21 days post-inoculation (PI). Tachyzoites were detected in lungs of all KO mice, and the T. gondii strains were successfully propagated in cell culture. The SW mice inoculated with tissues of wild turkeys remained asymptomatic, and tissue cysts were seen in the brains of infected mice when euthanatized in good health at 46 days PI; 1 of the 2 SW mice inoculated with the heart of 1 turkey died on day 26, and tachyzoites were detected in its lung. Genetic typing on DNA extracted from culture-derived tachyzoites using the PCR restriction fragment length polymorphism with 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) revealed that 4 isolates belonged to ToxoDB PCR-RFLP genotype #5 and 1 was genotype #216.