The influence of colostrum on infection of calves around 7 months of age with Schistosoma mattheei

Thirty red-legged partridges (Alectoris rufa), 5-month-old, were orally inoculated with oocysts of the OV-51/95 strain of Toxoplasma gondii. Birds were distributed into five groups and received, respectively, 10 (group A, 4 birds), 50 (group B, 14 birds), 10(2) (group C, 4 birds), 10(3) (group D, 4 birds) and 10(4) (group E, 4 birds) oocysts. One partridge from group B and one from group E died suddenly of acute toxoplasmosis at 7 day after inoculation (DAI) with demonstrable T. gondii in several tissues. The rest of birds remained clinically normal until killed at 44, 58, 65, 72, 79 or 100 DAI. Brain, heart, liver and skeletal muscle from these partridges were bioassayed individually in mice; T. gondii was demonstrated in all these tissues, except in heart of three birds inoculated, respectively, with 10, 50 and 10(2) oocysts. Lesions were not seen in histologic sections of tissues from surviving partridges. These results suggest that red-legged partridges are resistant to clinical toxoplasmosis.     

Serologic evaluation of cattle inoculated with Toxoplasma gondii: comparison of Sabin-Feldman dye test and other agglutination tests

Six cows and 6 calves were each inoculated with 100 or 100,000 Toxoplasma gondii oocysts. Serum samples were analyzed, using the Sabin-Feldman dye test (DT), indirect hemagglutination test, latex agglutination test, and the modified direct agglutination test (MAT). Antibody titers in cows were lower than in calves. In the cows, DT titers increased briefly during the first month after inoculation, after which the titers were negative; however, T gondii was isolated from the tissues of 4 cows. Indirect hemagglutination and latex agglutination titers were generally less than 1:256. The MAT titers increased to 1:1,024 during the first month after inoculation. In 5 of the 6 cows, the MAT titers persisted. The 6th cow had a preinoculation MAT titer of 1:2,000 for 3 to 6 months. Therefore, the DT was not useful in serologic surveys for T gondii in cattle; the MAT was the most sensitive test and may be useful in the diagnosis of T gondii infection in cattle.     

Experimental infection of budgerigars (Melopsittacus undulatus) with a low virulent K21 strain of Toxoplasma gondii

In total 53 budgerigars (Melopsittacus undulatus) were divided into six groups and orally infected with a suspension of oocysts of low virulent Toxoplasma gondii K21 strain in the doses of 10(2), 10(3), 10(4), 10(5) and 10(6), respectively. Blood was collected from the birds prior to the inoculation and then on days 10, 20 and 30 post infection. Latex-agglutination test (LAT) was used for the detection of antibodies in the inoculated birds. The infected birds showed no apparent signs of disease. The antibodies were found in all but two birds inoculated a dose of 10(2) oocysts. Haematological values remained unchanged after infection. T. gondii was isolated by bioassay in mice from all 37 birds fed 10(3) or more oocysts and 6 of 9 fed 10(2) oocysts. The results demonstrate that budgerigars are resistant to T. gondii infection.     

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.     

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.     

Serologic diagnosis of toxoplasmosis in experimentally infected pregnant goats and transplacentally infected kids

Eight pregnant goats were inoculated orally with 10 to 1,000 oocysts of Toxoplasma gondii at 83 to 102 days of gestation. Serum samples from the goats and from the kids born to them were analyzed, using the Sabin-Feldman dye test (DT), a commercially available modified agglutination test (MAT), and a latex agglutination test. Six of the does were observed for greater than 1 year; during this time, they delivered twice. All does developed DT and MAT antibody titers of greater than or equal to 1:2,048 within 29 days after inoculation, and the high titers persisted through the 2nd pregnancy; therefore, serologic results alone should not be relied on for the diagnosis of T gondii-induced abortion in does. On the other hand, all transplacentally infected kids had DT or MAT antibody titers of 1:2,048 before ingesting colostrum, indicating the usefulness of serologic evaluation of the fetus or stillborn kid in the diagnosis of abortion. Antibody was not found in the sera of noninfected kids born to Toxoplasma-infected does. The passively acquired colostral antibody declined by 5 months. Therefore, specific antibody found in adult goats is probably actively acquired. The commercially available MAT was simple, sensitive, and reliable for the diagnosis of caprine toxoplasmosis. The latex agglutination test needs further improvement, as titers rarely exceeded 1:256.     

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.     

First isolate of Toxoplasma gondii from arctic fox (Vulpes lagopus) from Svalbard

Cats are considered essential for the maintenance of Toxoplasma gondii in nature. However, T. gondii infection has been reported in arctic fox (Vulpes lagopus) from the Svalbard high arctic archipelago where felids are virtually absent. To identify the potential source of T. gondii, we attempted to isolate and genetically characterize the parasite from arctic foxes in Svalbard. Eleven foxes were trapped live in Grumant (78 degrees 11'N, 15 degrees 09'E), Svalbard, in September 2005 and 2006. One of the foxes was found to be seropositive to T. gondii by the modified agglutination test (MAT). The fox was euthanized and its heart and brain were bioassayed in mice for the isolation of T. gondii. All 10 mice inoculated with brain tissue and one of the five inoculated with heart developed MAT antibodies, and tissue cysts were found in the brains of seropositive mice. Two cats fed tissues from infected mice shed T. gondii oocysts. Genotyping using 10 PCR-RFLP markers and DNA sequencing of gene loci BSR4, GRA6, UPRT1 and UPRT2 determined the isolate to be Type II strain, the predominant T. gondii lineage in the world.     

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.     

Serodiagnosis of postnatally and prenatally induced toxoplasmosis in sheep

Nineteen pregnant (45 to 90 days of gestation) and 9 nonpregnant ewes were inoculated orally with 1,000 or 10,000 oocysts of Toxoplasma gondii. Pregnant ewes were euthanatized at days 14 (2 ewes), 21 (1 ewe), 23 (1 ewe), 28 (2 ewes), 35 to 42 (6 ewes), and 49 to 62 (6 ewes), and antibody titers in fetal and maternal sera were assayed, using the modified agglutination, latex agglutination, indirect hemagglutination, and dye tests. Although all ewes developed antibody titers of greater than or equal to 1,024 within 28 days after inoculation, fetuses were seronegative up to 28 days, using the modified agglutination test. Toxoplasma gondii antibodies were found in fetuses, using the modified agglutination and dye tests 35 days after ewes were inoculated. Latex agglutination and indirect hemagglutination tests were insensitive for detection of T gondii antibodies in ovine fetal sera. Toxoplasma gondii antibody titers in nonpregnant ewes were similar to those in pregnant ewes. Passively acquired T gondii antibodies from the colostrum decreased from 1,024 to less than 16 between 49 and 56 days of age in 1 lamb and between 62 and 106 days in its twin.     

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.     

Biologic and genetic comparison of Toxoplasma gondii isolates in free-range chickens from the northern Pará state and the southern state Rio Grande do Sul, Brazil revealed highly diverse and distinct parasite populations

The prevalence of Toxoplasma gondii in 84 free-range chickens (34 from the northern Pará state, and 50 from Rio Grande do Sul, the southern state) from Brazil, South America was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT), and found in 39 (46.4%) of 84 chickens with titers of 1:10 in one, 1:20 in two, 1:40 in four, 1:80 in seven, 1:160 in five, 1:320 in six, 1:640 in eight and > or =1:1280 in six. Hearts and brains of 45 chickens with titers of 1:20 or less were pooled and fed to two T. gondii-free cats. Hearts and brains of 39 chickens with titers of 1:10 or higher were bioassayed in mice. Feces of cats were examined for oocysts. One cat fed tissues from 31 chickens with titers of less than 1:10 from Rio Grande do Sul shed T. gondii oocysts. T. gondii was isolated by bioassay in mice from 33 chickens with MAT titers of 1:20 or higher. All infected mice from 10 isolates died of toxoplasmosis. All 34 isolates (15 from Pará, 19 from Rio Grande do Sul) were genotyped using 11 genetic markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2 and Apico. Eleven genotypes were revealed for Pará isolates and seven genotypes for Rio Grande do Sul. No genotype was shared between the two geographical locations. These data suggest that T. gondii isolates are highly diverse and genetically distinct between the two different regions in Brazil that are 3500 km apart.     

Concurrent Infection with Toxoplasma gondii and Feline Leukemia Virus: Antibody Response and Oocyst Production

Four adult cats (two testing positive and two negative for feline leukemia virus FeLV) were fed Toxoplasma gondii tissue cysts collected from the brains of mice. Two control cats (1 FeLV+, 1 FeLV-) were not fed cysts. The cats infected with T. gondii shed thousands of oocysts but remained clinically and physically normal, with hemograms and clinical chemistry values essentially unchanged irrespective of their FeLV status. Infection with FeLV did not increase the duration of oocyst shedding. At necropsy no significant lesions were found. T. gondii antibodies were detected by three serologic tests in the cats fed tissue cysts. The time necessary for an antibody response to T. gondii was not altered by the FeLV infection. Indirect hemagglutination (IHA) was the least reliable of the serologic tests studied; it detected antibodies later in the infection, and titers were less than in the other tests. Latex agglutination (LA) detected antibodies a few days before IHA, but titers were less than in modified direct agglutination (MAT). MAT detected antibodies earliest in the infection and also measured antibodies in aqueous humor and cerebrospinal fluid.     

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.     

Serologic prevalence of Toxoplasma gondii in horses slaughtered for food in North America.

Serum samples from 1788 horses slaughtered for food in North America were tested for antibodies to Toxoplasma gondii using the modified direct agglutination test (MAT). Antibodies to T. gondii were found by the MAT in 124 (6.9%) of 1788 sera; the titers were 1:20 (69 horses), 1:40 (37 horses), 1:80 (9 horses), and > or =1:160 (9 horses). A total of 339 selected horses were also tested by the Sabin-Feldman dye test (DT). Dye test antibodies were found in 54 horses with titers of 1:10 (29 horses) 1:20 (12 horses), 1:40 (4 horses) and 1:80 (9 horses). There was no correlation between the DT and the MAT.     

Toxoplasma gondii-induced abortion in dairy goats

Three of 8 goats on a Maryland farm aborted or had dystocia associated with toxoplasmosis during the winter of 1984. Doe 1 aborted a decomposed fetus 30 days before term. Modified agglutination test (MAT) antibody titers against Toxoplasma gondii were found in pleural fluid of the fetus (1:1,024) and in serum of doe (1:4,096 at 31 days after abortion). Doe 2 aborted a fetus 5 days before term; MAT antibody was found in the pleural fluid of the fetus (1:16,384) and in the doe's serum (1:4,096 on the day of abortion). Placenta from both does had foci of necrosis characteristic of toxoplasmosis, and T gondii was identified in lesions. Doe 3 had dystocia 7 days before term and a partially decomposed fetus was delivered by cesarian section; MAT was found in pleural fluid of the fetus (1:1,024) and in serum from the doe (1:4,096 on the day of abortion). Focal gliosis and calcification were seen in brain specimens from 2 of the 3 fetuses. None of the does produced milk after abortion. Two other does (No. 4 and 5) delivered apparently healthy kids transplacentally infected with T gondii; MAT in serum of both does was 1:4,096. Doe 4 delivered 3 kids; MAT titer in a serum from each kid 38 days after birth was 1:16,384. Doe 5 delivered 1 kid with a serum MAT titer of 1:1,024 at 38 days after birth. The 3 remaining does had MAT titers of 1:256, 1:16, and 1:16, and all delivered healthy kids. Epizootiologic evidence suggested that the does acquired T gondii infection from oocysts passed in feces of domestic cats on the farm. The MAT titers of 4 cats on the farm were 1:65,356; 1:1,024; 1:16; and 1:1,024.     

Evaluation of IFA, MAT, ELISAs and immunoblotting for the detection of anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs

The study evaluated the efficiency of diagnostic laboratory methods to detect anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs. 18-mixed breed pigs were used during the experiment; these were divided into two groups, G1 (infected group, n=10) and G2 (uninfected group, n=8). Infection was performed with 4 x 10(4) VEG strain oocysts at day 0 by the oral route in G1 animals. All pigs were euthanized at day 60, when retina, aqueous humour, and blood samples were collected. Anti-T. gondii antibody levels were assessed in serum (s) and aqueous humour (ah) by indirect immunofluorescence assay (IFA), modified agglutination test (MAT), m-ELISA (using crude membranes from T. gondii tachyzoites as antigen) and r-ELISA (using rhoptries from T. gondii tachyzoites as antigen). Polymerase chain reactions (PCR) of samples from the retina were performed by using Tox4 and Tox5 primers. Antibody titers of G1 animals ranged from 128 to 1024 and from 16 to 256 in serum and aqueous humour, respectively. There were differences in the correlation coefficients between IFA(s) x IFA (ah) (r=0.62, P=0.05), MAT(s) x MAT (ah) (r=0.97, P<0.0001); however, there was no significant difference between r-ELISA(s) x r-ELISA (ah) (r= 0.14, P=0.7). Antibodies present in serum and aqueous humour recognized similar antigens. Samples of retina were positive by PCR in 30% (3/10) of infected pigs. G2 animals remained without antibody levels and were PCR negative throughout the experiment. These results suggest that the use of a combination of tests and immunoblotting for paired aqueous humour and serum samples could improve the sensitivity and specificity for the diagnosis of ocular toxoplasmosis.     

Prevalence of Toxoplasma gondii in dogs from Sri Lanka and genetic characterization of the parasite isolates

The prevalence of Toxoplasma gondii in 86 street dogs from Sri Lanka was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT) and found in 58 (67.4%) of 86 dogs with titers of 1:20 in eight, 1:40 in four, 1:80 in 10, 1:160 in 22, 1:320 in six, 1:640 in five, and 1:1280 or higher in three. Hearts, tongues, and brains (either separately or pooled) of 50 dogs with MAT titers of 1:40 were selected for isolation of T. gondii by bioassays in mice. For bioassays, canine tissues were digested in pepsin and homogenates were inoculated subcutaneously into mice; the mice receiving canine tissues were examined for T. gondii infection. In all, T. gondii was isolated from 23 dogs. Interestingly, dog organs varied in their capacity to induce T. gondii infection in mice, muscles producing more positive results than the brain. The T. gondii isolates obtained from 23 seropositive dogs were PCR-RFLP genotyped using polymorphisms at 10 nuclear markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2, and an apicoplast marker Apico. Mixed infection with two genotypes was observed in one dog. Four genotypes were revealed, including three unique genotypes in addition to one belonging to the predominant Type III lineage. The 24 isolates were designated as TgDgSl 1-24.     

Isolation and characterization of Toxoplasma gondii strains from stray cats revealed a single genotype in Beijing, China

Cats are essential in the epidemiology of Toxoplasma gondii because they are the only hosts that can excrete the environmentally resistant oocysts in nature. This study was aimed to determine the seropositivity, distribution of genotypes and mouse virulence of T. gondii from stray cats in Beijing, China. A total of 64 serum samples, 23 feces and tissue samples were collected from stray cats in Beijing. Antibodies to T. gondii were assayed by the modified agglutination test (MAT). 57.8% (37/64) of these stray cats had titers of 1:20 or higher and were considered positive with infection. T. gondii oocysts were not found in feces of the 23 cats. Tissues of 23 cats were bioassayed in mice and 11 T. gondii isolates were obtained. The genotype of these isolates were identified by 11 PCR-RFLP markers, including SAG1, (3'+5')SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker, Apico. Only one genotype was identified. This genotype, designated as ToxoDB genotype #9, was previously reported in cats, pigs and human from Guangdong and Gansu provinces in China and animals from a few other countries. To determine mouse virulence of this lineage of parasites, one isolate was randomly selected and inoculated into BABL/c mice, the result showed that it is intermediately virulent to mice. These results indicated that an atypical, intermediately virulent T. gondii lineage is widespread in China. The high seropositivity of T. gondii in stray cats posts potential risk of transmission of the parasite to human population in the region.     

Seroprevalence and isolation of Toxoplasma gondii from free-range chickens from Espírito Santo state, southeastern Brazil

Prevalence of Toxoplasma gondii infection in 510 free-range (FR) chickens (380 from 33 small farms, and 130 from a slaughter house for FR chickens) from Espírito Santo state, southeastern Brazil, was investigated. Antibodies to T. gondii were sought using commercial indirect haemagglutination (IHAT, Imuno-HAI Toxo(?), Wama Diagnóstica, S?o Paulo, Brazil, cut-off 1:16) and the modified agglutination test (MAT, cut-off 1:25) tests. Attempts were made to isolate viable T. gondii from seropositive chickens by bioassay in mice. Pooled samples of brain, heart and quadriceps muscle of one thigh (total 40 g) from 64 chickens with IHAT titers of ≥ 1:16 were minced, digested in pepsin and bioassayed in mice. Antibodies to T. gondii were found in 40.4% (206/510) FR chickens by IHAT (titer ≥ 1:16) and 38.8% (198/510) by MAT (titer ≥ 1:25); concordance between IHAT and MAT was 81.6% (kappa index=0.614). Viable T. gondii was isolated (designated TgCkBr234-281) from 48 of 64 (75%) seropositive (IHAT titers ≥ 1:32) FR chickens. Most isolates of T. gondii were virulent for mice; 100% of mice inoculated with 44 of 48 isolates died of toxoplasmosis within 30 days post inoculation (p.i). An epidemiological investigation revealed that people living in rural areas have little knowledge about the parasite and about the risk of acquiring it from raw meat. Results indicated that the locally available IHAT was useful for screening of chicken sera for T. gondii antibodies.