Genetic and biologic characteristics of Toxoplasma gondii infections in free-range chickens from Austria

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in free-range chickens (Gallus domesticus) from 11 Bio-farms in Austria was determined. Antibodies to T. gondii assayed by the modified agglutination test (MAT) were found in 302 of 830 (36.3%) chickens with titers of 1:10 in 50, 1:20 in 69, 1:40 in 53, 1:80 in 40, 1:160 or higher in 90. Hearts of 218 chickens with MAT titers of 10 or higher were bioassayed individually in mice. Tissues from 1183 chickens were pooled and fed to 15, T. gondii-free cats. Feces of the cats were examined for oocysts; 11 cats shed T. gondii oocysts. T. gondii was isolated from 56 chickens by bioassay in mice. Thus, there were 67 isolates of T. gondii from these chickens. Genotyping of these 67 isolates using the SAG2 locus indicated that all 33 were Type II. Phenotypically and genetically these isolates were different from T. gondii isolates from Brazil. None of the isolates was virulent for mice. This is the first report of isolation of T. gondii from chickens from Austria.     

Characterization of Toxoplasma gondii isolates in free-range chickens from Chile, South America

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 85 free-range chickens (Gallus domesticus) from Chile was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 47 of 85 (55.3.9%) chickens with titers of 1:5 in six, 1:10 in four, 1:20 in four 1: 40 in three, 1: 80 in nine, 1: 160 in four 1:320 in nine, and 1: 640 or higher in eight. Hearts and brains of 47 chickens with titers of 1:5 or higher were pooled for each chicken and bioassayed in mice. Tissues from 16 seronegative (MAT<1:5) chickens were pooled and fed to one T. gondii-free cat. Feces of the cat were examined for oocysts but none was found based on bioassay of fecal floats in mice. Hearts and brains from seven seronegative (<1:5) were pooled and bioassayed in mice; T. gondii was not isolated. T. gondii was isolated by bioassay in mice from 22 chickens with MAT titers of 1:20 or higher. Genotyping of these 22 isolates using polymorphisms at the loci SAG1, SAG2, SAG3, BTUB and GRA6 revealed three genotypes. Seventeen isolates had type II alleles and four isolates had type III alleles at all loci. One isolate contained the combination of type I and III alleles. This is the first report of genetic characterization of T. gondii isolates from Chile, South America.     

Genetic and biologic characteristics of Toxoplasma gondii isolates in free-range chickens from Colombia, South America

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 77 free-range chickens (Gallus domesticus) from Colombia, South America was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 32 (44.4%) of 72 chickens with titers of 1:5 in 4, 1:10 in 3, 1:20 in 1, 1:40 in 1, 1:80 in 8, 1:160 in 8, 1:320 in 3, and 1:640 or higher in 4. Hearts and brains of 31 seropositive chickens were pooled and bioassayed in mice. Tissues from 32 (16+16) seronegative chickens were pooled and fed to two, T. gondii-free cats, and tissues from nine chickens without matching sera were fed to one T. gondii-free cat. Feces of cats were examined for oocysts. T. gondii oocysts were excreted by a cat that was fed tissues of 16 seronegative chickens. T. gondii was isolated by bioassay in mice from 23 chickens with MAT titers of 1:20 or higher. All infected mice from 16 of the 23 isolates died of toxoplasmosis. Overall, 82 (81.1%) of 101 mice that became infected after inoculation with chicken tissues died of toxoplasmosis. Genotyping of these 24 isolates using polymorphisms at the SAG2 locus indicated that seven T. gondii isolates were Type I, 17 were Type III, and none was Type II. Phenotypically, T. gondii isolates from chickens from Colombia were similar to isolates from Brazil but different from the isolates from North America; most isolates from chickens from Brazil and Colombia were lethal for mice whereas isolates from North America did not kill inoculated mice. Genetically, none of the T. gondii isolates from Colombia and Brazil was SAG2 Type II, whereas most isolates from chickens from North America were Type II. This is the first report of genetic characterization of T. gondii isolates from Colombia, South America.     

Biologic and genetic characteristics of Toxoplasma gondii isolates in free-range chickens from Nicaragua, Central America

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 98 free-range chickens (Gallus domesticus) from Nicragua was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 84 (85.7%) of 98 chickens with titers of 1:5 in 10, 1:10 in eight, 1:20 in seven, 1:40 in nine, 1:80 in 11, 1:160 in one, 1:200 in 27, 1:400 in six, 1:800 four, and 1:3200 in one bird. Hearts and brains of 32 chickens with titers of 1:10 or less were pooled and fed to three T. gondii-free cats. Hearts and brains of 66 chickens with titers of 1:20 or higher were bioassayed in mice. Feces of cats were examined for oocysts. The cat fed tissues from eight chickens with titers of 1:10 shed T. gondii oocysts. The two cats fed tissues of 24 chickens with titers of 1:5 or less did not shed oocysts. T. gondii was isolated by bioassay in mice from 47 chickens with MAT titers of 1:20 or higher. All infected mice from six isolates died of toxoplasmosis. Overall, 41 of 170 (24.1%) mice that became infected after inoculation with chicken tissues died of toxoplasmosis. Genotyping of these 48 isolates (47 from mice and 1 from pooled tissues) using polymorphisms at the loci SAG1, SAG2, SAG3, BTUB and GRA6 revealed eight genotypes. Six isolates had Type I alleles, three isolate had Type II alleles and six isolates had Type III alleles at all loci. Four isolates had mixed infections. Two isolates have a unique allele at SAG1 locus and combination of I and III alleles at other loci. The rest 27 isolates contained the combination of Type I and III alleles and were divided into four genotypes. More than one genotypes were often isolated in chickens from the same household, indicating multiple genotypes were circulating in the same environment. This may explain the high frequency of mixed infections observed. High rate of mixed infection in intermediate hosts such as chickens may facilitate genetic exchange between different parasite lineages in definitive feline hosts. This is the first report of genetic characterization of T. gondii isolates from Nicragua, Central America.     

Biologic and genetic characteristics of Toxoplasma gondii isolates in free-range chickens from Costa Rica, Central America

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 144 free-range chickens (Gallus domesticus) from Costa Rica was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 60 (40.1%) of 144 chickens with titers of 1:5 in 16, 1:10 in 5, 1:20 in 2, 1:40 in 3, 1:80 in 5, and 1:160 or higher in 29. Tissues of all chickens were bioassayed for T. gondii in mice or cats. Hearts and brains of 52 chickens with titers of 1:5 or higher and 16 chickens with doubtful titers were pooled and bioassayed in mice. Tissues from 76 chickens with MAT titers of 1:10 or less were pooled and fed to three T. gondii-free cats. Fecal floats of cats were bioassayed orally in mice but were negative for T. gondii oocysts. T. gondii was isolated by bioassay in mice from 32 chickens with MAT titers of 1:10 or higher. All infected mice from 4 of the 32 isolates died of toxoplasmosis. Genotyping of these 32 isolates using polymorphisms at the loci SAG1, SAG2, SAG3, BTUB and GRA6 revealed five genotypes. Five isolates had type I alleles and one isolate had type III alleles at all loci. The rest 26 isolates contained the combination of type I and II or I and III alleles and were divided into three genotypes. None was found to have genotype II alleles at all five loci. This is the first report of genetic characterization of T. gondii isolates from Costa Rica, Central America.     

Detection of Toxoplasma gondii in free-range chickens in China based on circulating antigens and antibodies

Toxoplasma gondii is widely distributed in humans and other animals including domestic poultry throughout the world, but the data on prevalence of T. gondii in free-ranged (FR) chickens in People's Republic of China (PRC) are limited. In the present study, the seroprevalence of T. gondii infection in FR chickens was investigated in 13 provinces/municipalities of China during the period from January to June 2010. A total of 1173 serum samples were collected and assayed for T. gondii circulating antigens (TCA) and antibodies (TCAb) using enzyme linked immunosorbent assay (ELISA) technique. Out of this number, 199 samples were TCA positive (16.97%), 226 samples were TCAb positive (19.27%), 69 samples were positive for both TCA and TCAb (5.88%), and the total seropositive rate was found in 356 of 1173 (30.36%). The results of the present survey indicated that infection with T. gondii in FR chickens is widely spread in China.     

Genotyping of Toxoplasma gondii isolates from chickens from India

The present study was undertaken to isolate and genotype Toxoplasma gondii from free-range chickens (Gallus domesticus) from villages in Maharashtra and Tamil Nadu states of central and south India, respectively. Blood, heart, and brain from a total of 741 chickens were examined for T. gondii infection. Antibodies to T. gondii, as assayed with the modified agglutination test (MAT >or = 1:5) were found in 133 (17.9%) chickens. Hearts and brains of 186 chickens were bioassayed in mice. Additionally, hearts and/or brains of most of the seronegative (MAT < 1:5) chickens were fed to 20 T. gondii-free cats, while 32 seropositive chickens (MAT 1:5) were fed to 3 cats. T. gondii was not isolated from any of the chickens by mouse bioassay. Five of the cats that were fed seronegative chickens shed oocysts, while isolates were not obtained from any of the other cats fed seropositive chickens. These five isolates, along with the two that were previously isolated in India through cat bioassay, were genetically analyzed. Genotyping using the SAG 2 locus indicated that two isolates were type II and five were type III. Microsatellite analysis revealed allelic differences between and within the lineages. This is the first report of genetic characterization of any T. gondii isolate from India.     

Genetic diversity of Toxoplasma gondii isolates from chickens from Brazil

Until recently, Toxoplasma gondii was considered clonal with very little genetic variability. Recent studies indicate that T. gondii isolates from Brazil are genetically and biologically different from T. gondii isolates from USA and Europe. In the present study, we retyped 151 free range chicken isolates from Brazil including 117 newly isolated samples from 11 geographically areas (Alagoas, Bahia, Ceará, Maranh?o, Paraná, Pernambuco, Rio de Janeiro, Rio Grande do Norte, S?o Paulo, Sergipe, and Rondonia) and 34 previously reported isolates from the very north (Pará) and the very south (Rio Grande do Sul). Ten PCR-RFLP markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico were used to genotype all isolates. Overall analysis of 151 T. gondii isolates revealed 58 genotypes. Half (29/58) of these genotypes had single isolate and the other half of the genotypes were characterized with two or more isolates. Only 1 of 151 isolates was clonal Type I strain and 5 were clonal Type III strains. Two isolates had mixed infections. Clonal Type II strain was absent. One strain was Type II at all loci, except BTUB. The results confirm high genetic diversity of T. gondii isolates from Brazil.     

High prevalence of Toxoplasma gondii in a commercial flock of chickens in Israel, and public health implications of free-range farming

Little is known of the prevalence of Toxoplasma gondii in commercially raised chickens. In the present study, the prevalence of T. gondii in 96 free-range chickens (Gallus domesticus) from a commercial farm in Israel was assessed. Blood, heart, and brain from each chicken were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT > or = 1:5), were found in 45 of the 96 chickens. Hearts and brains of seropositive (MAT > or = 1:5) chickens were bioassayed in mice. Additionally, hearts and brains of 51 seronegative (MAT < 1:5) chickens were bioassayed in two T. gondii-free cats. T. gondii was isolated from 19 of the 45 (42.2%) seropositive chickens by bioassay in mice. Both the cats fed tissues pooled from seronegative chickens shed T. gondii oocysts. Tachyzoites and tissue cysts of all 21 isolates of T. gondii from chickens were avirulent for mice. Seventeen of the 19 isolates genotyped were found to be type II, and 2 were type III. Understanding of the sources of infection on such farms could be the key to the development of better prevention strategies.     

Isolation and molecular characterization of Toxoplasma gondii from chickens and ducks from Egypt

The prevalence of Toxoplasma gondii in free range chickens is a good indicator of the prevalence of T. gondii oocysts in the environment because chickens feed from the ground. In the present study, prevalence of T. gondii in 121 free range chickens (Gallus domesticus) and 19 ducks (Anas sp.) from a rural area surrounding Giza, Egypt was assessed. Blood, heart, and brain from each animal were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT), were found in 49 (40.4%) chickens in titers of 1:5 in 11, 1:10 in four, 1:20 in four, 1:40 in eight, 1:80 in 10, and 1:160 or more in 12 chickens. Antibodies were found in three ducks each with a titer of 1:80. Hearts and brains of seropositive (MAT > or = 1:5) chickens and ducks were bioassayed in mice. Additionally, hearts and brains of seronegative (MAT<1:5) animals were bioassayed in T. gondii-free cats. T. gondii was isolated from 19 of 49 seropositive chickens (one with a titer of 1:5, two with a titer of 1:20, one with a titer of 1:40, five with a titer of 1:80, three with a titer of 1:160, and seven with a titer of > or = 1:360). One cat fed tissues pooled from 15 seronegative chickens shed T. gondii oocysts, while two cats fed tissues of 34 seronegative chickens did not shed oocysts. T. gondii was isolated from one of the seropositive ducks by bioassay in mice. The two cats fed tissues from 16 seronegative ducks did not shed oocysts. Genotyping of 20 chicken isolates of T. gondii using the SAG 2 locus indicated that 17 isolates were type III and three were type II. The duck isolate of T. gondii was type III. The mice inoculated with tissue stages of all 21 isolates of T. gondii from chickens and ducks remained asymptomatic, indicating that phenotypically they were not type I because type I strains are lethal for mice. Infections with mixed genotypes were not found.     

Toxoplasma gondii infection in Polish farmed mink

The aim of this study was to determine the seroprevalence of Toxoplasma gondii antibodies in Polish farmed mink according to way of feeding as well as to confirm the role of toxoplasmosis in reproductive losses in mink farms. The serological examinations were carried out on 961 mink randomly selected from 12 Polish farms. Blood sera were examined for the presence of T. gondii antibodies with the use of the latex agglutination test. The examinations for the presence of T. gondii in organ tissues were performed on five neonatal mink kits with the use of immunofluorescence method. In total 133 (13.9%) out of 961 examined mink had T. gondii antibodies. In large farms the seropositivity was lower (2.9%), than in small farms (26.33%) (P < 0.001). Significant difference was found in seroprevalence according to way of feeding. In farms feeding fish, percentage of seropositivity was lower (2.2%), than in farms based on non-frozen slaughter offal (43.4%). Titres of T. gondii antibodies were usually lower than 120 IU/ml. Using the immunofluorescence method, T. gondii was detected in impression smears from liver and brain of two neonatal mink kits derived from one seropositive female.     

Prevalence of Toxoplasma gondii infection in raccoons.

Serum samples from 427 raccoons (93 from Pennsylvania, 45 from New Jersey, 72 from South Carolina, 68 from Virginia, 30 from Iowa, and 119 from Ohio) were evaluated for Toxoplasma gondii antibodies in dilutions of 1:25, 1:50, and 1:500. The distribution of T gondii antibody titers was less than 1:25 for 212 raccoons (49.6%), 1:25 for 34 raccoons (7.9%), 1:50 for 117 raccoons (27.4%), and greater than or equal to 1:500 for 64 raccoons (14.9%). Tissue cysts were seen in the liver, and tachyzoites were in the brain of a raccoon with abnormal neurologic signs and concurrent infection with canine distemper virus. Organisms in the liver were stained with anti-T gondii serum, and the raccoon had a T gondii titer of 1:160 in the agglutination test.     

Detection of antibodies to Toxoplasma gondii in stillborn piglets in Argentina.

Fetal fluids of 738 stillborn piglets from three swine farms in Argentina were examined for antibodies to Toxoplasma gondii. Antibodies were detected in 15 samples at a 1:20 dilution in the indirect fluorescent antibody test and 10 samples were positive in the modified agglutination test (MAT) at a dilution of 1:25; four of these samples had a MAT titer of > or = 1:100. This survey indicates a low rate of congenital T. gondii infection in stillborn pigs in Argentina.     

Genotyping of Toxoplasma gondii isolates from free range chickens in the Pantanal area of Brazil

The aim of this paper was to genetically characterize Toxoplasma gondii isolates from free range chickens in regions of Brazilian territory in the state of Mato Grosso do Sul (MS) where T. gondii strains have never been studied. In total, T. gondii isolates from 22 free range chickens were included in this study. Fifty chickens from Eldorado, thirty from Rio Verde and ten from Aquidauana were sampled between January and April 2007. In relation to the genetic diversity of T. gondii isolates from chickens in MS, the magnitude of the diversity in the isolates sampled in this study was comparable to the overall diversity in a composite data set. These 22 isolates in MS revealed 11 genotypes, whereas the 321 isolates ever genotyped in Brazil have revealed 95 genotypes. The values of Simpson's Diversity Index for the whole population of T. gondii isolates in Brazil, the whole population of T. gondii isolates from chickens in Brazil and the population surveyed in this study were 0.97, 0.95 and 0.90, respectively. Seven of the 11 genotypes revealed from chicken isolates from MS are newly described genotypes and six of them each have a single isolate. In conclusion, the results obtained from isolates in MS corroborate previous studies on T. gondii isolates in Brazil, thus confirming their diversity and atypicality. Nonetheless, the applicability of PCR-RFLP markers for epidemiological inferences remains controversial.     

Serological survey of Toxoplasma gondii infection in domestic cats from northeastern Portugal

Cats are very important hosts in the epidemiological cycle of Toxoplasma gondii, a zoonotic protozoan parasite that can infect humans and many other animal species worldwide. We report a serological survey of antibodies to T. gondii in domestic cats from northeastern Portugal, by means of the modified agglutination test. Three cats had titres of 20 (3.9%), 18 had titres of 40 (23.7%) and 55 animals had titres of > or =800 (72.4%). Results of three seropositive kittens with less than 4 months were not considered for determining the seroprevalence of infection, which was found to be 35.8% (73/204). Differences in the seroprevalence levels were not statistically significant between males (35.6%) and females (36.0%) or pure non-European (26.7%) and European or mixed-breed cats (39.6%). Animals aged 36-71 months and 72-180 months had the highest seroprevalences of infection, i.e. 51.7% and 51.2%, respectively, which significantly differ from the values observed in cats with 2-11 months (14.6%) and 12-35 months (26.3%). Infection levels were also significantly different between cats that lived totally indoors (7.7%) and those that had access to outdoors (45.4%), as well as between cats living alone (13.8%) and those that had contact with other cats (39.4%). Seroprevalence values in cats fed only commercial canned or dried food (22.9%) and animals whose diet included raw or undercooked viscera and/or meat (53.5%) were also significantly different. Furthermore, considering only 108 cats, differences of seropositivity to T. gondii were significant between feline immunodeficiency virus infected and non-infected animals, but this was not observed for feline leukaemia virus. Age, habitat and diet were identified as risk factors for the feline T. gondii infection by logistic regression analysis. Some control measures are suggested based on these findings.     

Prevalence of antibodies to Sarcocystis neurona, Toxoplasma gondii and Neospora caninum in horses from Argentina.

Sera from 76 horses from Argentina were examined for antibodies to Sarcocystis neurona, Toxoplasma gondii and Neospora caninum. Antibodies to S. neurona were found in 27 (35.5%) of 76 horses using immunoblots with culture derived merozoites as antigen. Antibodies to T. gondii were found in 10 (13.1%) of 76 horses by using the modified agglutination test with formalin-fixed tachyzoites and mercaptoethanol; titers were 1:25 (two horses), 1:50 (six horses), 1:100 (two horses), and 1:200 (one horse). Antibodies to N. caninum were not found in any of the 76 horses by the use of N. caninum agglutination test. This is the first report of S. neurona infection in horses in Argentina.     

Seroprevalence of Toxoplasma gondii in sows from slaughterhouses and in pigs from an indoor and an outdoor farm in Argentina

The objective of the present study was to determine the prevalence of Toxoplasma gondii antibodies from slaughter sows and from pigs raised at an indoor and an outdoor swine farm. Serum samples were obtained from 230 slaughter sows belonging to 83 farms distributed in 5 provinces. Blood samples were collected monthly from pigs of different ages from an intensive management indoor farm (farm 1). A cross-sectional study was carried-out from an outdoor farm (farm 2). All sera were tested for T. gondii antibodies by the modified agglutination test (MAT), using formalin-fixed tachyzoites as antigen. An antibody titer > or =1:25 was considered positive. Antibodies to T. gondii were detected in 87 (37.8%) of 230 sows sera. Distribution among provinces was: 37.1% from Santa Fe, 62.8% from Buenos Aires, 3.3% from San Luis, 58.7% from La Pampa and 24% from Córdoba. Four of 88 (4.5%) serum samples from farm 1 had antibodies to T. gondii and none of the negative pigs seroconverted. However, 45 of 112 samples from farm 2 were positive (40.2%) with the following distribution: sows 100%; nursery 40%; growers 13.8% and fatteners 20%. It is concluded that the prevalence of T.gondii antibodies among sows seems to be quite variable. T. gondii prevalence was related to the facilities and management of the farm.     

Prevalence of Toxoplasma gondii infection in Belgian house cats

Five hundred and sixty seven sera of healthy house cats aged 3 months to 7 years, were examined for the presence of anti-toxoplasma antibodies by indirect immunofluorescence assay and compared to SAG1 and TLA enzyme linked immunosorbent assays as alternative test. Twenty-five percent of cats tested positive for IgG and/or IgM. Seroprevalence increased with age from 2% below 12 months of age up to 44% at age 7. Sensitivities of SAG1 and TLA ELISA were 84.1% and 88.6%, respectively. Peak levels in seroprevalence were correlated to increased IgG titers in TLA ELISA. Our results suggest that T. gondii infections are common in house cats and that there is a high chance for a negative cat to seroconvert in its second life-year.     

Seroprevalence of Toxoplasma gondii infection in Norwegian dairy goats

Background

Toxoplasma gondii is a major problem for the sheep industry as it may cause reproduction problems. The importance of T. gondii in Norwegian goat herds is uncertain, but outbreaks of toxoplasmosis in dairy goat farms have been recorded. The aim of this study was to describe the prevalence of T. gondii infection in Norwegian dairy goats by using serology.

Findings

Goat serum originally collected as part of two nationwide surveillance and control programmes between 2002 and 2008 were examined for T. gondii antibodies by using direct agglutination test. In total, 55 of 73 herds (75%) had one or more serologically positive animals, while 377 of 2188 (17%) of the individual samples tested positive for T. gondii antibodies.

Conclusions

This is the first prevalence study of T. gondii infection in Norwegian goats. The results show that Norwegian goat herds are commonly exposed to T. gondii. Nevertheless, the majority of goat herds have a low prevalence of antibody positive animals, which make them vulnerable to infections with T. gondii during the gestation period.