Transplacental toxoplasmosis in a reindeer (Rangifer tarandus) fetus

Toxoplasma gondii infection was diagnosed in a full term stillborn reindeer (Rangifer tarandus) fetus. The fetus had encephalitis and placentitis associated with T. gondii. Tissue cysts were identified histologically in sections of brain and tachyzoites were present in placenta and the myocardium. Protozoa in the brain, heart, and placenta stained positively with T. gondii antibodies, but not with Neospora caninum antibodies in an immunohistochemical test. The dam of the fetus had a 1:12,800 titer to T. gondii in the modified agglutination test employing whole tachyzoites and mercaptoethanol. This is the first confirmed report of T. gondii infection in reindeer.     

Toxoplasmosis in black-faced kangaroos (Macropus fuliginosus melanops)

An epizootic of toxoplasmosis among captive black-faced kangaroos (Macropus fuliginosus melanops) is reported. Eight of 25 adult kangaroos had antibodies to Toxoplasma gondii. Serologic data indicated recent exposure to T. gondii in six kangaroos. Two kangaroos had high T. gondii antibody titers (greater than or equal to 16,384) in the modified agglutination test and their infants died when less than 7 months old. Toxoplasma gondii tachyzoites were found in several organs of one infant kangaroo (joey) that died at about 82 days of age and numerous cysts were seen in skeletal muscles of the other joey that died at about 7 months of age. Adult kangaroos had subclinical infections. The modified agglutination test and the dye test were more sensitive than the indirect hemagglutination and latex agglutination tests for the detection of T. gondii antibodies in kangaroo sera.     

Evaluation of a recombinant MIC3 based latex agglutination test for the rapid serodiagnosis of Toxoplasma gondii infection in swines

The entire gene encoding microneme protein 3 (MIC3) from Toxoplasma gondii was cloned into the plasmid pGEX-KG and subsequently expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. The recombinant MIC3 (rMIC3) was purified and evaluated in a latex agglutination test (LAT) as the diagnostic antigen for the detection of antibodies to T. gondii in pig sera. The specificity, stability, and reproducibility of the test were examined. No agglutination was found when the sensitized latex beads were mixed with phosphate-buffered saline (PBS), borate-buffered saline (BBS), normal saline, and negative serum samples. There was no cross-reactivity with the standard positive sera of other pathogens. But intense agglutination occurred with T. gondii antibody positive serum samples. In our study, the coincidence rate of tested positive-sera of the LAT with rMIC3-sensitized latex particles and the ELISA with rSAG1 was up to 92.8%, T. gondii specific antibodies were detected by the LAT in all piglets that were experimentally infected with T. gondii tachyzoites from 8 to 42 days after infection. Our results indicated that the rMIC3 based latex agglutination test appears to be suitable for the detection of T. gondii antibodies at the early stage of infection.     

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.     

Toxoplasmosis as a suspected cause of abortion in a Greenland muskox (Ovibos moshatus wardi).

Toxoplasma gondii tachyzoites were seen in the placenta of a late-term aborted Greenland muskox (Ovibos moschatus wardi) fetus in a captive herd at the San Francisco Zoo. The organism stained with anti-T. gondii polyclonal rabbit serum but not with anti-Neospora caninum serum. The dam had a Toxoplasma titer of > or =1:3,200 at the time of abortion and in each of the previous 3 yr (modified agglutination test). The muskox is a new host record for T. gondii.     

Toxoplasma gondii antibodies in exotic wild felids from Brazilian zoos.

Serum samples from 37 captive exotic felids in 12 zoos from six Brazilian states were assayed for antibodies to Toxoplasma gondii by the modified agglutination test using formalin-fixed whole tachyzoites. Titers greater than or equal to 1:20 were considered positive. Antibodies to T. gondii were found in 24 of 37 (64.9%) felids, including one European lynx (Lynx lynx), two jungle cats (Felis chaus), two servals (Leptailurus serval), two tigers (Panthera tigris), three leopards (Panthera pardus), and 14 of 27 lions (Panthera leo). This is the first serologic analysis for T. gondii infection in exotic wild felids from Brazilian zoos.     

Toxoplasmosis in captive Bennett's wallabies (Macropus rufogriseus) in Argentina

Wallabies and other Australian marsupials are among the most susceptible species to Toxoplasma gondii. Fatal generalized toxoplasmosis was diagnosed in two captive 3 year-old female Bennett's wallabies (Macropus rufogriseus) from Argentina (w 1 and w 2) with a history of sudden death. Both animals had internal joeys which died 2 days after their mothers. Serologically, both females and one adult male without clinical signs from the same enclosure (w 3) had antibody titers for T. gondii>or=800 by the modified agglutination test (MAT); another adult male (w 4) was negative (MAT titer<25). Microscopically, tachyzoites were observed associated to non-suppurative meningoencephalitis, hepatitis, myositis, myocarditis and severe enteritis in hematoxylin and eosin stained sections from both w 1 and w 2. Immunohistochemically, parasites in heart, brain and liver sections of both female wallabies reacted with T. gondii antiserum. T. gondii was isolated from brain tissues of w 1 and w 2 by bioassay in mice and by culture in bovine monocytes and both isolates were cryopreserved. Genomic DNA was isolated from tachyzoites grown in cultures derived from both animals. The primer pair B22/B23 specific for T. gondii produced 115bp amplicons on poliacrylamide electrophoretic gels. Stray cats were suspected as the possible source of infection. Not all infected macropods were ill, showing that the infection may be asymptomatic and is not always fatal. A vertical infection could not be proved in the joey from w 2. As far as we know, this is the first confirmed report of toxoplasmosis in Bennet's wallabies in Argentina.     

磺胺氯吡嗪处理弓形虫速殖子抑制消减文库的构建和初步分析

为了研究和探索磺胺氯吡嗪抗弓形虫的作用机理,构建磺胺氯吡嗪处理弓形虫的抑制性消减文库.将刚地弓形虫(Toxoplasma gondii)RH株速殖子腹腔注射(10000个/只鼠)感染昆明系小鼠,大量收集并纯化速殖子.用250 mg/mL磺胺氯吡嗪PBS溶液和PBS分别浸泡速殖子,37℃恒温箱孵育2h,分别提取虫体的总RNA和mRNA,以PBS处理的正常虫体cDNA为驱动组,以药物处理虫体cDNA为实验组,构建药物处理前后弓形虫速殖子的抑制性消减文库,对部分阳性克隆进行测序和序列分析.结果显示,提取的虫体总RNA纯度高,合成的双链cDNA经RsaⅠ酶切反应较完全;从消减文库中随机选择100个阳性克隆进行PCR扩增,其插入片段长度在200～750 bp之间,文库的重组率达93％以上;随机挑取30个阳性克隆进行测序,获得11个单一序列,Blast分析显示有7个单一有效EST序列与已知蛋白质相似性较高.结果表明,已成功构建磺胺氯吡嗪处理弓形虫速殖子的抑制性消减cDNA文库,并对部分阳性克隆进行了初步分析,为深入研究磺胺氯吡嗪的抗弓形虫机制和寻找药物作用的关键靶分子奠定了基础.     

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.     

Occurrence of anti-<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> antibodies in female cattle in south-west of Iran

Toxoplasmosis, caused by Toxoplasma gondii, is a significant disease in livestock and humans. In Iran, studies shows that T. gondii infection in humans is relatively high and prevalence is associated mainly with consumption of undercooked meat or meat products. We have examined 450 serum samples from female cattle distributed over all Ahvaz, the center of Khouzestan province, south-west of Iran. IgG antibodies to T. gondii were assayed by the modified agglutination test using whole tachyzoites of T. gondii, and found in 71 (15.77%) of 450 cattle with titers of 1:25 in 38, 1:50 in 18, 1:100 in 11, 1:200 in three and 1:400 in one. Titers of antibodies were decreased in cattle over 2 years old. These results indicate that T. gondii infection in cattle of Khouzestan is relatively considerable, but not very high and consumption of beef may be a source of infection for humans in south-west of Iran.     

Prevalence of antibodies to Neospora caninum and Toxoplasma gondii in gray foxes (Urocyon cinereoargenteus) from South Carolina

Little is known about the epidemiology of Neospora caninum in wild mammal populations. It has been suggested that a sylvatic cycle exists for N. caninum. Dogs and potentially other canids are a definitive host for N. caninum. The present study was done to determine the prevalence of antibodies to N. caninum in a population of gray foxes (Urocyon cinereoargenteus) from a nonagricultural setting in South Carolina. We also determined the prevalence of antibodies to Toxoplasma gondii in these animals. Antibody levels were measured in direct agglutination tests using either N. caninum or T. gondii formalin-fixed tachyzoites as antigen. Four (15.4%) of the 26 gray foxes had titers to N. caninum. Titers to N. caninum were low being 1:25 in three gray foxes and 1:50 in the fourth gray fox. Antibodies to T. gondii were observed in 16 (61.5%) gray foxes. Titers to T. gondii were usually >1:50 and two gray foxes had titers of 1:1600. Results of this study indicate that gray foxes have more exposure to T. gondii than to N. caninum in this environment.     

Porcine abortion outbreak associated with Toxoplasma gondii in Jeju Island, Korea

This report deals with the acute onset of an abortion outbreak and high sow mortality in one pig herd consisted of 1,200 pigs and 120 sows on Jeju Island, Korea. Affected pregnant sows showed clinical signs, including high fever, gradual anorexia, vomiting, depression, recumbency, prostration, abortion, and a few deaths. Four dead sows, five aborted fetuses from the same litter, and 17 sera collected from sows infected or normal were submitted to the Pathology Division of the National Veterinary Research and Quarantine Service for diagnostic investigation. Grossly, hepatomegaly and splenomegaly were observed in sows. Multiple necrotic foci were scattered in the lungs, liver, spleen, and lymph nodes. Microscopically, multifocal necrotizing lesions and protozoan tachyzoites were present in the lesions. Tachyzoites of Toxoplasma (T.) gondii were detected immunohistochemically. Latex agglutination showed that the sera of 7 of 17 (41.2%) sows were positive for antibody to T. gondii. The disease outbreak in this herd was diagnosed as epizootic toxoplasmosis. To our knowledge, this is the first report of porcine toxoplasmosis with a high abortion rate and sow mortality in Korea.     

Unexpected oocyst shedding by cats fed Toxoplasma gondii tachyzoites: in vivo stage conversion and strain variation

Tachyzoites, bradyzoites (in tissue cysts), and sporozoites (in oocysts) are the three infectious stages of Toxoplasma gondii. The prepatent period (time to shedding of oocysts after primary infection) varies with the stage of T. gondii ingested by the cat. The prepatent period (pp) after ingesting bradyzoites is short (3-10 days) while it is long (18 days or longer) after ingesting oocysts or tachyzoites, irrespective of the dose. The conversion of bradyzoites to tachyzoites and tachyzoites to bradyzoites is biologically important in the life cycle of T. gondii. In the present paper, the pp was used to study in vivo conversion of tachyzoites to bradyzoites using two isolates, VEG and TgCkAr23. T. gondii organisms were obtained from the peritoneal exudates (pex) of mice inoculated intraperitoneally (i.p.) with these isolates and administered to cats orally by pouring in the mouth or by a stomach tube. In total, 94 of 151 cats shed oocysts after ingesting pex. The pp after ingesting pex was short (5-10 days) in 50 cats, intermediate (11-17) in 30 cats, and long (18 or higher) in 14 cats. The strain of T. gondii (VEG, TgCKAr23) or the stage (bradyzoite, tachyzoite, and sporozoite) used to initiate infection in mice did not affect the results. In addition, six of eight cats fed mice infected 1-4 days earlier shed oocysts with a short pp; the mice had been inoculated i.p. with bradyzoites of the VEG strain and their whole carcasses were fed to cats 1, 2, 3, or 4 days post-infection. Results indicate that bradyzoites may be formed in the peritoneal cavities of mice inoculated intraperitoneally with T. gondii and some bradyzoites might give rise directly to bradyzoites without converting to tachyzoites.     

CpG-oligodeoxynucleotides enhance porcine immunity to Toxoplasma gondii

Protection against a challenge infection with Toxoplasma gondii VEG strain oocysts was examined in pigs after vaccination with T. gondii RH strain tachyzoites with or without a porcine specific synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs. Six groups of pigs were immunized with incomplete Freund's adjuvant (IFA) and either vehicle, tachyzoites alone or in combination with three different doses of CpG ODN or with CpG ODN alone. Protection from challenge was significantly (P < 0.05) improved in pigs vaccinated using CpG ODN as an adjuvant with tachyzoites compared to all other groups. The CpG ODN tachyzoite-immunized pigs also had higher serum parasite specific IgG antibody, no clinical signs of disease, and 52% had no demonstrable tissue cysts after the challenge infection. These data indicate that CpG ODN is a potential safe and effective adjuvant for the T. gondii RH strain vaccine in pigs.     

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.     

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.     

Neospora caninum associated with septic peritonitis in an adult dog

A 7-year-old, male neutered Rhodesian Ridgeback dog was referred to the University of California-Davis Veterinary Medical Teaching Hospital with a 4-month history of peritonitis and episodic abdominal discomfort, lethargy, and weakness. Marked abdominal distension with a prominent fluid wave was noted on physical examination. Cytologic analysis of the abdominal fluid indicated a septic exudate with mixed bacteria and many protozoal zoites. Differentials for the identity of the protozoal zoites included Toxoplasma gondii, Sarcocystis neurona, and Neospora caninum. Indirect latex agglutination antigen testing, standard indirect fluorescent antibody testing, and PCR analysis were performed to identify the zoites. The dog's serum antibody titer for N caninum tachyzoites was 1:20,480, known polysera to N caninum reacted against zoites in the abdominal fluid, and PCR analysis of the abdominal fluid was positive for the presence of a known gene of N caninum. Based on the morphologic, immunologic, and molecular findings, the zoites were identified as N caninum. It remains unclear how the tachyzoites gained access to the peritoneal cavity. To the authors' knowledge, there are no reports of free N caninum in abdominal fluid of any species.     

Dermatitis in a dog associated with an unidentified Toxoplasma gondii-like parasite

Protozoal dermatitis was diagnosed in a 6-year-old female Great Dane dog from Rio de Janeiro, Brazil. The dog died because of a chronic illness with an Ehrlichia-like organism. Numerous apicomplexan parasites were identified histologically in the section of dermal lesions. The protozoan reacted with Toxoplasma gondii polyclonal rabbit serum but not with Neospora caninum or Sarcocystis neurona antibodies. Ultrastructurally, the protozoa was not T. gondii because it had schizont-like structures with merozoites arranged around a prominent residual body, and the merozoites had several rhoptries with electron-dense contents; rhoptries in T. gondii tachyzoites are electron-lucent and a residual body is not found in groups of tachyzoites. This is the first report of unidentified T. gondii-like protozoa in the skin of a dog.     

Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and Sarcocystis canis-like infections in marine mammals

Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and S. canis are related protozoans that can cause mortality in many species of domestic and wild animals. Recently, T. gondii and S. neurona were recognized to cause encephalitis in marine mammals. As yet, there is no report of natural exposure of N. caninum in marine mammals. In the present study, antibodies to T. gondii and N. caninum were assayed in sera of several species of marine mammals. For T. gondii, sera were diluted 1:25, 1:50, and 1:500 and assayed in the T. gondii modified agglutination test (MAT). Antibodies (MAT > or =1:25) to T. gondii were found in 89 of 115 (77%) dead, and 18 of 30 (60%) apparently healthy sea otters (Enhydra lutris), 51 of 311 (16%) Pacific harbor seals (Phoca vitulina), 19 of 45 (42%) sea lions (Eumetopias jubatus) [corrected] 5 of 32 (16%) ringed seals (Phoca hispida), 4 of 8 (50%) bearded seals (Erignathus barbatus), 1 of 9 (11.1%) spotted seals (Phoca largha), 138 of 141 (98%) Atlantic bottlenose dolphins (Tursiops truncatus), and 3 of 53 (6%) walruses (Odobenus rosmarus). For N. caninum, sera were diluted 1:40, 1:80, 1:160, and 1:320 and examined with the Neospora agglutination test (NAT) using mouse-derived tachyzoites. NAT antibodies were found in 3 of 53 (6%) walruses, 28 of 145 (19%) sea otters, 11 of 311 (3.5%) harbor seals, 1 of 27 (3.7%) sea lions, 4 of 32 (12.5%) ringed seals, 1 of 8 (12.5%) bearded seals, and 43 of 47 (91%) bottlenose dolphins. To our knowledge, this is the first report of N. caninum antibodies in any marine mammal, and the first report of T. gondii antibodies in walruses and in ringed, bearded, spotted, and ribbon seals. Current information on T. gondii-like and Sarcocystis-like infections in marine mammals is reviewed. New cases of clinical S. canis and T. gondii infections are also reported in sea lions, and T. gondii infection in an Antillean manatee (Trichechus manatus manatus).     

First Portuguese isolate of Neospora caninum from an aborted fetus from a dairy herd with endemic neosporosis

Neospora caninum was isolated from the brain of an aborted 4-month-old fetus from a dairy cow herd with endemic neosporosis in Porto, Portugal. The fetal brain homogenate was inoculated interperitoneally first into outbred Swiss Webster mice given dexamethasone and then the peritoneal exudates from these mice was co-inoculated with mouse sarcoma cells in the peritoneal cavity of mice given dexamethasone. N. caninum tachyzoites were seen in peritoneal exudate of the second passage. Tachyzoites from the peritoneal exudate reacted positively with anti-N. caninum antibodies and not with anti-Toxoplasma gondii antibodies and contained N. caninum specific DNA. This Portuguese isolate of N. caninum has been successfully maintained in cell culture. The dam of the aborted fetus had an antibody titer of 1:10240 in the Neospora agglutination test (NAT). Antibodies to N. caninum were found in 76 of 106 cows from this herd in titers of 1:40 in 31, 1:80 in 22, > or =1:160 or more in 23 in the Neospora agglutination test. This is the first isolation of a viable N. caninum-like parasite from any host in Portugal.