Toxoplasma-like sporozoa in an aborted equine fetus

Multifocal areas of necrosis and infiltrations of mononuclear cells were seen in lung specimens of an equine fetus aborted 2 months before term. Extracellular and intracellular protozoa were seen in the alveolar tissue. Individual organisms were 4 microns by 2.5 microns, and cyst-like structures were 25 microns by 18 microns. Organisms did not stain with periodic acid-Schiff or by use of the immunoperoxidase and peroxidase-antiperoxidase method for Toxoplasma gondii. Twelve days after abortion, the mare had serum antibody titer of less than 1:10 against T gondii.     

Congenital sporozoan encephalomyelitis in a calf

Protozoan encephalomyelitis was diagnosed post mortem in a five-day-old Friesian calf which had shown nervous signs from birth. The lesion was a subacute necrotising multifocal encephalomyelitis associated with protozoan bodies (6 X 6 microns to 16 X 30 microns). Ultrastructurally these bodies corresponded to apicomplexan meronts composed of eight to 89 merozoites which reproduced by internal budding to form paired daughter cells (endodyogeny). The merozoites were indistinguishable from Toxoplasma gondii; they reacted weakly with anti-Sarcocystis species serum and did not cross react with anti-T gondii serum. The generic identity of the sporozoan was not established, but it is unlikely to have been either Sarcocystis species or T gondii.     

Neospora-like encephalomyelitis in a calf: pathology, ultrastructure, and immunoreactivity

Protozoal encephalomyelitis was diagnosed in a 3-day-old calf that was stunted, weak, and recumbent. Grossly, the calf had contracted tendons in the forelegs, a slightly doomed skull, a porencephalic cyst in the cerebellum, ulcerative esophagitis, and abomasitis. Histologically, there was a multifocal nonsuppurative encephalomyelitis with clusters of protozoal tachyzoites and numerous protozoal cysts. The porencephalic cyst and gastrointestional lesions appeared to be unrelated to the protozoal infection and were suggestive of a concurrent bovine virus diarrhea infection. A few groups of protozoal tachyzoites and numerous tissue cysts were found in neuropile, particularily in neurons of the spinal cord. By light microscopy, smaller tissue cysts were found in the brain (majority from 14 to 20 microns) than in the spinal cord (majority from 20 to 48 microns). The cyst walls ranged in thickness from less than 1 micron to a maximum of 2 microns wide. Bradyzoites contained PAS-positive slender bradyzoites (5-8 x 1-2 microns). Tissue cysts reacted positively to anti-Neospora caninum sera; but unlike N. caninum, they were positive to 2 of 4 antisera against Toxoplasma gondii and to antisera to H. hammondi. Ultrastructurally, tissue cysts closely resembled a Neospora-like organism, including the finding of interneuronal protozoal cysts, thick cyst walls, a lack of micropores in the bradyzoites, and the presence of numerous micronemes oriented perpendicular to the pellicle. Ultrastructural features in the calf protozoan that have not been reported for N. caninum in dogs included the presence of numerous tubulovesicular structures in the cyst ground substance and bradyzoite vesicles that contained small vesicular structures and short, flat membrane segments.(ABSTRACT TRUNCATED AT 250 WORDS)     

Congenital Neospora caninum infection in a calf with spinal cord anomaly

Neospora caninum was identified in a calf with spinal cord anomaly in Australia. The calf was full-term and born dead. The caudal cervical and cranial thoracic segments of the spinal cord of the calf were asymmetric because of marked unilateral reduction of ventral gray matter and focal cavitation. Mild focal disseminated nonsuppurative encephalomyelitis was associated with N caninum tissue cysts. Tissue cysts were 16 to 35 microns X 10 to 27 microns, and the cyst walls were 1 to 3 microns thick. In an immunohistochemical test, the parasite stained with N caninum serum but not T gondii serum.     

Additional details of the life history of the chicken coccidium Eimeria mitis Tyzzer, 1929

Four asexual generations of Eimeria mitis were identified. The first three developed above the epithelial cell nuclei, but the fourth developed above and below. Meronts measured 13.8 x 16.4 microns, 16.1 x 16.4 microns, 12.1 x 14.6 microns, and 9.5 x 12.4 microns, respectively, of generations 1, 2, 3, and 4. They matured at 36, 67, 72, and 88 hr postinoculation (PI) and contain 20-24, 16-20, 10-14, and 7-10 merozoites, respectively. Merozonts measured 7.2 x 1.9 microns, 8.5 x 2.5 microns, 9.6 x 2.0 microns, and 6.75 x 2.75 microns, respectively. The first two types of meronts were deep in the crypts and epithelial cells. The third and fourth types of meronts were along the side and tip of the villi. Gametocytes developed from third and fourth generation. Gamonts were usually below the nuclei of the epithelial cells. Parasitism was primarily in the ileum, ceca, and rectum and also in the yolk-sac diverticulum.     

Eimeria capricornis n.sp., E. nihonis n.sp., E. naganoensis n.sp., and E. kamoshika n.sp. (Protozoa: Eimeriidae) from the Japanese serow, Capricornis crispus

Eimeria capricornis n.sp., E. nihonis n.sp., E. naganoensis n.sp. and E. kamoshika n.sp. were detected from the Japanese serow, Capricornis crispus. The oocysts of these species were ovoid to ellipsoid, measuring 48.02 +/- 0.53 x 33.93 +/- 0.35 microns, 29.73 +/- 0.45 x 21.01 +/- 0.29 microns, 20.04 +/- 0.05 x 15.77 +/- 0.04 microns and 30.02 +/- 4.24 x 14.58 +/- 2.03 micron, respectively. In all the species the micropyle was observed, but the micropylar cap and oocyst residuum were not present. The polar granules were observed in the oocysts of E. capricornis. The sporocysts of the above species were spherical to elongate ovoid, measuring 15-23 x 8-13 microns, 14-16 x 7-10 microns, 11-13 x 6-8 microns and 8-10 x 6-7 microns, and the sporulation time was 6, 3, 3 and 6 days, respectively. The sporocyst residuum and refractile body were seen in all the species. In the sporocysts of E. naganoensis and E. kamoshika tiny Stieda body was seen, but not in the other two species. These four Eimeria species were not infective to goats.     

First isolation of Sarcocystis hominis from cattle in Japan.

Sarcocystis hominis was first isolated from slaughtered cattle raised in Japan. Cysts were 1,220-4,460 x 80-384 microns in size and their wall was 3 to 6 microns thick and appeared radially striated in the histopathological sections because of the presence of palisade-like villar protrusions on the surface. The protrusions were 3.1-4.3 x 0.7-1.1 microns in size and had many microtubules in the core. Two cynomolgus monkeys, Macaca fascicularis, fed with the Sarcocystis cysts began to pass sporocysts, which measured a size of 14.3-15 x 9.5-10 microns, in the feces 10 days after ingestion.     

Toxoplasmosis in sheep III. Further evaluation of the ability of a live Toxoplasma gondii vaccine to prevent lamb losses and reduce congenital infection following experimental oral challenge

The aim of this study was to compare the ability of a live incomplete strain (Strain 48) and a live complete strain (Strain 89) of Toxoplasma gondii to protect against abortion and congenital infection following an oral challenge of T. gondii oocysts. Sixty-nine two-tooth ewes were immunised pre-tupping with live Strain 48 of T. gondii tachyzoites and seventy ewes were immunised with Strain 89. Eighty-two serologically negative ewes served as controls. At mid-pregnancy half of the ewes were challenged orally with T. gondii oocysts (2x10(5)/ewe). The ewes vaccinated with Strain 48 were significantly (p<0.05) protected against the effects of experimental challenge and the rate of congenital infection was also significantly (p<0.15) reduced. The ewes vaccinated with Strain 89 were also significantly (p<0.05) protected. The serological response to challenge as measured by both the Dye test and the Indirect Haemagglutination test varied considerably between the two vaccinated groups.     

Exfoliative colpocytology: a method for the diagnosis of Capuchin monkey filariasis?

During vaginal fluid examinations (Papanicolaou) to study the physiological sexual cycle of Cebus sp., abundant Dipetalonema gracile microfilariae (110-160 microns x 4-5 microns, without a sheath) were encountered in the genital fluid, but not in peripheral blood. Considering the great difficulty in diagnosing this obscure parasitosis, exfoliative colpocytology was found to be an efficient diagnostic.     

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.     

The frequency and geographic distribution of Giardia infections in ruminants in Switzerland

In various districts (Cantons) of Switzerland 815 calves (a few days up to 6 months old), 382 lambs and 20 young goats (both groups 1-6 months old) were selected randomly for a single coprological examination (flotation method) for Giardia infection. In average 26.6% of the calves, 29.8% of the lambs and 4 of 20 young goats excreted Giardia cysts. In 9 Cantons the percentages of cyst excretors among the calves varied between 15 and 32, but these differences were not significant. Further there were no significant differences in the frequency of cyst excretion between calves of 3 different breeds and 2 age groups (up to 3 months old and 3 to 6 months). The intensity of cyst excretion was high and varied in random samples between 4.1 x 10(3) and 3.0 x 10(5) cysts per g of faeces in calves and between 2.2 x 10(3) and 1.6 x 10(5) in lambs. In 5 of 7 farms where calves and lambs were maintained simultaneously both animal species were Giardia infected. As expected, trophozoites of Giardia isolates from calves and lambs belonged to the Giardia duodenalis type and could not be differentiated on a morphological basis. Giardia cysts from calves (average measurements: 13.7 x 9.1 microns) and lambs (13.8 x 9.2 microns) were indistinguishable both morphologically and morphometrically. The results indicate that Giardia infections are frequent and geographically widely distributed in calves and lambs in Switzerland.     

Characterization of Toxoplasma gondii isolates from free range chickens from Paraná, Brazil

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 40 free range chickens (Gallus domesticus) from a rural area surrounding Paraná, Brazil 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 16 chickens. Hearts and brains of seropositive (MAT> or =1:5) chickens were bioassayed in mice. Additionally, hearts and brains of seronegative (MAT<1:5) chickens were bioassayed in two T. gondii-free cats (12 chickens per cat). T. gondii was isolated from 13 of 16 (81%) seropositive chickens. Of the two cats fed tissues pooled form seronegative chickens, one shed T. gondii oocysts. Nine of the 13 T. gondii isolates killed 100% of infected mice. The T. gondii isolate from the cat was also virulent for mice. Genotyping of 13 chicken isolates of T. gondii using the SAG2 locus indicated that seven isolates were type I and six were type III; three of these type III isolates killed all infected mice suggesting that all strains virulent for mice are not type I. The isolate from the feces of the cat fed chicken tissues was type I.     

Effect of Feline Immunodeficiency Virus Infection on Toxoplasma gondii-specific Humoral and Cell-mediated Immune Responses of Cats with Serologic Evidence of Toxoplasmosis

Serum samples from 89 cats with serologic evidence of toxoplasmosis were identified by using an enzyme-linked immunosorbent assay (ELISA) that detected Toxoplasma gondii -specific immunoglobulin M (IgM) or T. gondii -specific immunoglobulin G (IgG). Concurrent feline immunodeficiency virus (FIV) infection was detected in 36 cats using an ELISA for detection of FIV-specific IgG. The majority of the cats in both the FIV-seropositive and FIV-seronegative groups were male and >5 years of age. FIV-seropositive cats were more likely to have T. gondii IgM titers without IgG ( P > 0.05) or any T. gondii IgM titer ( P > 0.05) than were FIV-seronegative cats. FIV-seronegative cats (1328) had a higher T. gondii IgG geometric mean titer than did FIV-seropositive cats (724) and were more likely to have T. gondii IgG titers 1:2048 than were FIV-seropositive cats ( P > 0.05). Cats with serologic evidence of both T. gondii and FIV infections had persistent T. gondii IgM titers for >12 weeks. Lymphoblast transformation in response to concanavalin A, T. gondii -specific intracellular antigens, and T. gondii -specific secretory antigens was compared in T. gondii seropositive and FIV-seronegative cats, cats with serologic evidence of T. gondii infection alone, and cats with serologic evidence of concurrent FIV and T. gondii infections. Lymphocytes from all but one cat in the FIV-seropositive group responded to concanavalin A. Whereas lymphocytes from FIV-seronegative cats with serologic evidence of toxoplasmosis responded to T. gondii -specific antigens, four of five of the FIV-seropositive cats with concurrent serologic evidence of toxoplasmosis did not.     

Toxoplasmosis in sheep. II. The ability of a live vaccine to prevent lamb losses after an intravenous challenge with Toxoplasma gondii

Two-tooth ewes (n=48) were immunized pre-tupping with a live Toxoplasma gondii vaccine. At midpregnancy these ewes were challenged intravenously with 1 x 105 live T. gondii tachyzoites. The strain of T. gondii used for vaccination was an incomplete strain that did not produce oocysts. It was derived by continuous twice weekly passage in mice. The lambing percentage for ewes immunized with the live vaccine was significantly higher (P<0.001 normal score) than non-vaccinated control ewes. However, vaccination did not prevent foetal or placental infection. The serological response to vaccination and challenge was measured by both the Dye test and the Indirect Haemagglutination test. No significant relationship between titre of antibody and protection in the vaccinated ewes was observed.     

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.     

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.     

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.     

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

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

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.     

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.