Role of endogenous transplacental transmission in toxoplasmosis in sheep

To investigate the potential role of endogenous transplacental transmission of Toxoplasma gondii, 31 seropositive ewes presumed to be persistently infected with the parasite and 15 seronegative ewes were mated and monitored throughout pregnancy and lambing. Antibody titres were determined in precolostral sera from the liveborn lambs and in thoracic fluid from the dead lambs. A PCR for the B1 gene of T gondii was applied to the placentas from all the ewes and to the brains of the stillborn lambs. Samples of brain, lung, liver, spleen and heart from the dead lambs were examined by histopathology. No evidence of toxoplasmosis was detected by histopathology or PCR in any of the samples, but low titres of antibody to T gondii were detected in two liveborn, healthy offspring of a seropositive ewe by the immunofluorescent antibody test (3.2 per cent of pregnancies and 4.1 per cent of lambs in the seropositive group). Antibody to specific antigens of T gondii was demonstrated in sera from these two lambs by Western blotting.     

Toxoplasma gondii-induced abortion in sheep

Twenty-nine of 200 (14.5%) ewes on a farm in Cobleskill, NY aborted or had dead lambs during the lambing seasons of 1985 and 1986. Thirteen of 15 ewes that aborted had high Toxoplasma gondii antibody titers (1,024), via the modified agglutination test, and T gondii was isolated from the tissues of 1 fetus. In the 1987 lambing season, 5 ewes aborted, but not because of T gondii infection. Toxoplasma gondii antibodies were detected in 73.8% of sera obtained from 592 ewes in January 1987, indicating enzootic toxoplasmosis on this farm. Seropositivity increased with age; 40.2% of 1-year-old ewes had detectable antibody vs 89.2% of 2-year-old ewes.     

Epizootiologic investigations on a sheep farm with Toxoplasma gondii-induced abortions

During the lambing season of 1983/1984, 8 of 44 purebred Hampshire ewes on a farm in Knoxville, Md had reproductive problems. In at least 4 of these ewes, the problem was attributed to toxoplasmosis. Necrosis and Toxoplasma gondii tachyzoites were found in placental specimens from 3 ewes. Agglutinating antibody to T gondii, at a titer of 1:80, was found in pleural fluids of both fetuses aborted from 1 ewe; this ewe had an antibody titer of 1:6,400 at the time of abortion. In another ewe, the diagnosis was confirmed by the isolation of T gondii from the placenta and 1 of her lambs. Of numerous free-roaming adult cats on the farm, 16 were trapped, euthanatized, and examined for T gondii. Agglutination antibody to T gondii, at titers of 1:4 to 1:64, was found in serum samples from all the cats. Toxoplasma gondii was isolated from the brain and skeletal muscles of 9 of the cats, and from the feces of 1 cat. Blood samples obtained from all 78 sheep on the farm 6 months after the episode of abortion were examined for antibody to T gondii. Agglutinating antibody titers to T gondii were less than 1:16 in 46 sheep, 1:16 in 16, 1:64 in 12, 1:256 in 2, 1:024 in 1, and 1:4,096 in 1. Analyses of serologic data in sheep of various age groups suggested that the Toxoplasma infection was acquired sporadically, probably from feed contaminated with oocysts.     

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.     

Intrauterine transmission of ovine progressive pneumonia virus

Ovine fetuses, newborn lambs, and ovine colostrum were examined for ovine progressive pneumonia virus. The lambs and colostrum were also examined for specific antibody. Virus was isolated from 1 fetus, from 2 newborn lambs, and from most samples of colostrum. The fetus was about 100 days old and was carried by a seronegative ewe in contact with seropositive sheep. Both newborn lambs were carried by seropositive ewes. One lamb was dead at birth; the other lamb was normal and had not nursed. Antibody specific for the virus was present in the colostrum of 12 of 14 seropositive ewes and in the serum of 8 of 11 lambs that had nursed seropositive ewes, but not in the serum of lambs that had not nursed.     

Placental transfer of specific antibodies during ovine congenital toxoplasmosis

The passage of non-Toxoplasma antibodies from dam to fetus through damaged placenta was studied in sheep inoculated with Toxoplasma gondii. Six ewes were inoculated with chicken globulins and Leptospira bacterins 2 months before oral inoculation with Toxoplasma gondii oocysts. Ewes were euthanatized between 42 and 62 days after T gondii inoculation. Antibody titers against chicken globulins, Leptospira spp, Haemonchus contortus, Sarcocystis spp, and T gondii were measured in the maternal and fetal sera. All ewes became infected with T gondii and had grossly visible necrotic foci in the placentas, and T gondii antibodies were found in the fetuses and the ewes. Appreciable amounts of antibodies to Haemonchus contortus, Sarcocystis sp, Leptospira spp, and chicken globulins did not cross the placental barrier. Seemingly, serologic examination of the fetus was reliable for the diagnosis of ovine congential toxoplasmosis.     

Response of immune and susceptible ewes to infection with Toxoplasma gondii

Inocula containing 75, 250 or 1000 Toxoplasma gondii tissue cysts were used to infect seronegative gimmers and seropositive ewes in the fourth month of pregnancy. The seronegative gimmers developed typical toxoplasma infections at all dose levels. Four of them aborted and the surviving lambs showed rising indirect haemagglutination titres in the first two to three months of life indicating congenital infection. The seropositive ewes showed no response to challenge, all their lambs survived and there was no serological evidence of congenital infection. Indirect haemagglutination titres in the seropositive ewes remained unchanged throughout the experiment, titres in the gimmers rose sharply from the 10th day after infection and by three months were the same as those in the ewes.     

Survey of Toxoplasma antibodies among sheep in western United States

A survey was conducted to determine the prevalence of toxoplasma antibodies among breeding ewes and among lambs slaughtered for food in western United States. Each serum was tested by the indirect hemaglutination method, using microtiter technique. Agglutination (greater than or equal to 2 +) at the 1:64 dilution was considered to be a positive reaction. Of 2,164 ewes from 18 flocks tested in California, 523 (24%) were seropositive for Toxoplasma gondii, with prevalence rates among flocks ranging from 4 to 51%. In 9 of those flocks, 1,495 ewes were stratified by whether ewes had lambed or were barren. On an overall basis, the antibody prevalence was similar (about 25%) in both groups, but there was a significant difference (P less than 0.05) in 1 flock in which 30% of the nursing ewes were seropositive, compared with 21% of the barren ewes. Of 1,056 market lambs from 19 lots tested, 85 (8%) were seropositive. The antibody prevalence in lambs tested at slaughter in California, by state of origin, were: Oregon, 11/51 (22%); Nevada, 32/159 (20%); Idaho, 12/147 (5%), and California, 30/699 (4%).     

Studies on the response of ewes to live chlamydiae adapted to chicken embryos or tissue culture.

Ewes infected before gestation with chicken embryo or tissue culture adapted chlamydial strain B-577 were challenge inoculated with the homologous strain at four to 18 weeks of gestation. The ewes responsed with group specific complement fixing antibody titers of 1:8 to 1:256 by the second week after initial infection. A secondary antibody response in the surviving challenge inoculated ewes occurred at the time of lambing and reached titers of 1:32 to 1:256 by the second week after parturition. Group specific complement fixing antibodies did not appear to play a significant role in resistance to chlamydial infection. Ewes infected with the chicken embryo adapted strain B-577 excreted chlamydiae in their feces 60 days after inoculation. However, chlamydiae were not recovered from feces of ewes infected with the tissue culture adapted strain B-577. Placentas of ewes challenge inoculated by the intravenous route were consistently infected. Chlamydiae were recovered from placentas, some fetuses and lambs. In two instances when challenge inoculation was given by the intramuscular route, infection was detected only by the direct fluorescent antibody method.     

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.     

The prevalence of antibodies against Toxoplasma gondii among hospitalized animals and stray dogs.

Hospitalized animals and stray dogs were serologically tested for antibodies against Toxoplasma gondii. In addition, the data were examined for the possibility of toxoplasmosis infection being associated with the clinical diagnosis and with the discharge status (alive vs. dead). Among 1056 hospitalized animals, 17 (20%) of 86 cats, 112 (14%) of 804 dogs, 34 (26%) of 133 horses and 6 (18%) of 33 cattle had serological evidence of infection with T. gondii. Only 22 (6%) of 342 young (median age = one year) stray dogs were seropositive. The difference in antibody prevalence between hospitalized and stray dogs was thought to be due to age and dietary factors. Of 249 dogs grouped by clinical diagnosis, there was significantly (p less than 0.01) higher prevalence of seropositives among dogs with diseases of the kidney or with adrenocortical hyperfunction than among dogs hospitalized for other diseases. Of 19 dogs with diseases of the kidney and 12 with adrenocortical hyperfunction 37% and 42%, respectively, were seropositive.. There was higher risk of being discharged from the hospital dead among seropositive dogs, cattle and horses than among seronegative animals of the same species. The exception was cats, where of 69 seronegative cats 29% were dead at discharge and where of 17 seropositive cats 18% were dead at discharge. The possible effects of stress due to hospitalization need further research.     

Economic and public health considerations of congenital toxoplasmosis in lambs

Congenital toxoplasmosis was diagnosed in a flock of Hampshire sheep in South Dakota. The 80 ewes produced 144 lambs, 30 of which were born dead; toxoplasmosis was diagnosed in 11 of the dead lambs. The remaining 114 lambs grew normally, but 68 (40.3%) had agglutinating Toxoplasma gondii antibodies. Modified agglutination test T gondii antibody titers for 114 lambs were: less than 100 (n = 46), 64 (n = 2), 256 (n = 1), 1,024 (n = 12), and greater than or equal to 4,096 (n = 53). Tissues of 8 seropositive lambs were bioassayed for T gondii tissue cysts. Toxoplasma gondii was isolated from the tongue and lamb chops of 7, heart of 3, and legs of lamb of all 8 lambs.     

Correlation of tissue infection and serologic findings in pigs fed Toxoplasma gondii oocysts

Each of thirteen 6-week-old pigs was inoculated per os with 10,000 sporulated oocysts of Toxoplasma gondii. By postinoculation day (PID) 13, pigs were seropositive by the indirect fluorescent antibody test. Beginning on PID 13 and every 7 days thereafter through PID 97, 1 pig was killed and 6 tissues were examined for T gondii. Of the 13 pigs, 11 were infected, including the 1st pig killed on PID 13, although none of the pigs had gross lesions of toxoplasmosis. Tissues harboring T gondii most frequently were the heart and brain; organisms were detected less frequently in the longissimus muscles, diaphragm, and liver. Toxoplasma gondii was not detected in the bronchial lymph nodes. There was good correlation between antibody and presence of T gondii in these pigs. One additional pig, maintained as a noninfected control, remained seronegative and had no evidence of infection when killed on PID 97.     

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.     

Toxoplasma gondii antibodies in California swine.

In a serologic survey for Toxoplasma gondii in 891 swine from 20 California counties, 478 (54%) were seronegative, 153 (17%) were nonspecific reactors, and 260 (29%) were seropositive. Most of the titers (81%) were low, ie, from 1:64 to 1:128, but a few (2%) were high, ie, 1:1024 or greater.     

Pilot project for eradicating maedi-visna in Walliser blacknose sheep

Maedi-Visna is a lentiviral disease of sheep with a worldwide distribution. The transmission of the virus occurs primarily via colostrum and milk from the infected ewe to its newborn lamb but also horizontally between sheep. The most obvious clinical symptoms are progressive dyspnea and emaciation. In this prospective study an eradication based on serological testing and removing of seropositive animals was performed in 24 flocks of sheep of the breed "Walliser Schwarznasenschafe" leading to a reduction of the seroprevalence from 36% to 1% within two years. The control group consisted of 21 flocks of sheep. Lambs of seropositive ewes had a 7.6 times higher risk to seroconvert within their first two years of life compared to those of seronegative ewes. The dynamics of the spread of the infection were studied in birth cohort groups. Cohort animals of seropositive ewes showed an obvious trend to seroconvert slowly. Seropositive ewes had a significantly lower reproduction rate and their lambs suffered from significantly higher death and lower growth rates, probably due to a reduced milk production, resulting in economic losses.     

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

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

Evaluation of ewe and lamb immune response when ewes were supplemented with vitamin E

Fifty-two Targhee twin-bearing ewes were used in a factorial arrangement of treatments to investigate the role of supplemental vitamin E (vit E); 0 (NE) vs 400 IU of vit E x ewe x (-1)d(-1) (E) and parainfluenza type 3 (PI3) vaccination; none (NP) vs PI3 vaccination (P) in immune function. Parainfluenza type 3 vaccination was used to evoke an immune response. Ewes receiving PI3 were vaccinated at 49 and 21 d before the expected lambing date. Ewes receiving vit E were orally dosed daily, 32 to 0 d before lambing. Blood was collected from ewes at the time of the initial PI3 vaccination and 4 h postpartum. Blood was collected from lambs (n = 104) at 3 d postpartum. Ewe and lamb sera were analyzed for anti-PI3 antibody titers, immunoglobulin G (IgG) titers, and vit E concentrations. Colostrum was collected 4 h postpartum and analyzed for IgG. The model for ewe and lamb analysis included the main effects of vit E and PI3, sex (lambs model only), and their interactions. No interactions were detected (P > 0.20) for any ewe or lamb variables. Serum anti-PI3 titers were greater (P < 0.01) in P ewes and their lambs than NP ewes and their lambs. Serum vit E concentrations were greater (P < 0.01) in E ewes and their lambs than NE ewes and their lambs. Colostral IgG titers and serum anti-PI3 titers did not differ (P > 0.20) between E and NE ewes. Serum IgG titers in E ewes and their lambs did not differ (P > 0.15) from IgG titers in NE ewes and their lambs. Lamb anti-PI3 titers did not differ (P = 0.76) between lambs reared by E and NE ewes. These results indicate that, although supplemental vit E to the ewe increased lamb serum vit E concentration, it had no effect on measures used in this study to assess humoral immunity in the ewe or passive immunity to the lamb.     

Persistence of granulocytic Ehrlichia infection during wintertime in two sheep flocks in Norway

Granulocytic Ehrlichia infection in sheep is common in Norway in areas with Ixodes ricinus. In this study, 2 sheep flocks that had been grazing on I. ricinus infested pastures the previous season, were blood sampled after being housed indoors for nearly 6 months during wintertime. Thirty animals from each flock were examined for granulocytic Ehrlichia infection in the peripheral blood by blood inoculation studies, stained blood smear evaluation, polymerase chain reaction (PCR) analysis and serology (IFA-antibodies). The animals were sampled twice within a three-week period, the first time before and the second time after lambing. Two sheep in one flock were found Ehrlichia positive by both blood smear evaluation and PCR before lambing, and 3 sheep were found positive after lambing; 2 by blood smear examination and 3 by PCR. In the other flock, no sheep was found infected before lambing, but 2 ewes were found positive after lambing by both blood smear evaluation and PCR. In the first flock, 87% of the animals were found seropositive before lambing, and the mean antibody titre (log10 +/- SD) to E. equi was 2.45 +/- 0.401. In the second flock, 40% were found seropositive before lambing, and the mean antibody titre was 1.93 +/- 0.260. Seroprevalence and mean antibody titre in these 2 flocks were significantly different (p < 0.001). The present study indicates that sheep may be a reservoir host for granulocytic Ehrlichia infection from one grazing season to the next under natural conditions in Norway.     

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.