Cutaneous toxoplasmosis in a female Japanese cat

A 16-year-old female Japanese cat was presented with a single mammary-gland nodule approximately 3 cm in diameter. Histologically, the nodule consisted of necrotizing granulomatous panniculitis, vasculitis, and mastitis, and contained free and clustered protozoal organisms. The organism was present in the cytoplasm of macrophages, fibroblasts, endothelial cells, and mammary-gland epithelia. The organism was positive for anti- Toxoplasma gondii and anti- Neospora caninum antibodies. Electron microscopy showed single and grouped tachyzoites, with morphologic features similar to those of T. gondii. Polymerase chain reaction and deoxyribonucleic acid sequence analysis was consistent with T. gondii infection. This is the first report of cutaneous toxoplasmosis in a Japanese cat.     

Feline cutaneous toxoplasmosis: a case report

A 9-year-old female, domestic short hair cat was presented with sudden onset of polyuria/polydipsia, and hundreds of cutaneous nodules. Prior to referral, the cat had had four skin nodules that were treated with steroids. The four skin nodules then multiplied to form more than 100 ulcerated and nonulcerated nodules located all over the trunk. Clinical evaluation revealed hypothermia and respiratory distress. Cytology from both skin nodules and bronchoalveolar lavage showed macrophages and small organisms whose shape and size were indicative of Toxoplasma spp., or similar organisms. Feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) serology results were negative. The cat was seropositive for Toxoplasma (IgG 1 : 640) and Neospora (1 : 80) infections. The cat died soon after referral. Necropsy revealed pyothorax, necrotic/purulent pneumonia, haemorrhagic spots on kidneys and mesentery. Histopathology from skin nodules showed diffuse, deep necrotic dermatitis/panniculitis, vasculitis and disseminated free and grouped protozoa. The parasites were found in lungs, spleen, kidneys and liver. Immunohistochemistry on skin tissue with anti-Toxoplasma gondii and Neospora caninum antibodies gave positive results with both. Electron microscopy showed single and grouped tachyzoites with morphological features of T. gondii, often within macrophages. Samples of cutaneous nodules and bronchoalveolar fluid were examined by a polymerase chain reaction (PCR) assay for detecting apicomplexa coccidia. PCR results were consistent only with T. gondii infection. Therefore, immunohistochemistry positivity for N. caninum was considered a cross-reaction and a diagnosis of cutaneous and visceral toxoplasmosis was made.     

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.     

Clinicopathologic diagnosis of cutaneous toxoplasmosis in an Angora cat

This report deals with a case of cutaneous toxoplasmosis in a 2 year-old female Angora cat. Cutaneous lesions were characterized by prescapular ulcers and hyperemic nodules in the skin of the inguinal and dorsosacral regions. A skin biopsy sample was collected from the lesioned area and processed for histopathologic examination and immunoperoxidase test using Toxoplasma gondii and Neospora caninum specific antibodies. Toxoplasma gondii immunopositive reactions were detected in keratinocytes and dermal macrophages while no immunoreactivity was detected for N. caninum. The case of cutaneous toxoplasmosis was further confirmed by PCR analysis using T. gondii B1 gene-specific primers. In conclusion, we report the first case of cutaneous toxoplasmosis in Angora cats.     

Toxoplasmosis in Tammar wallabies (Macropus eugenii) in the Budapest Zoo and Botanical Garden (2006-2010)

Smaller macropodid species (commonly referred to as wallabies) are extremely susceptible to toxoplasmosis: in most cases, infection with Toxoplasma gondii leads to death within a short time. Between June 2006 and July 2010, T. gondii was detected by immunohistochemical examination in six Tammar wallabies (Macropus eugenii) that died in the Budapest Zoo and Botanical Garden; in another four specimens histopathology revealed T. gondii-like organisms (which could not be differentiated from Neospora caninum solely by morphology), and in another 11 animals toxoplasmosis as the possible cause of death could not be excluded. The current zoo population of 12 Tammar wallabies was tested for T. gondii IgG antibodies by the modified agglutination test (MAT), with negative results. We suppose that most of the deaths were due to acute toxoplasmosis resulting from a recent infection.     

Occurrence of Neospora caninum and Toxoplasma gondii infections in ovine and caprine abortions

Neospora caninum and Toxoplasma gondii are closely related cyst-forming apicomplexan parasites identified as important causes of reproductive failure in cattle and small ruminants, respectively. Protozoan abortion in small ruminants is traditionally associated with T. gondii, but the importance of N. caninum remains uncertain. The aim of this study was to investigate the presence of N. caninum and T. gondii infections in abortion cases in small ruminants submitted for diagnosis. For this purpose, 74 ovine and 26 caprine aborted foetuses were recovered from different areas in Spain. Foetal histopathology was used to detect the presence of protozoal-associated lesions in brain. The presence of N. caninum and T. gondii was confirmed by PCR. Protozoal infection was detected in 17 out of 100 (17%) foetuses examined by at least one of the diagnostic techniques used. Lesions suggestive of protozoal infection were observed in 10.8% (8/74) and 15.4% (4/26) of the ovine and caprine abortions respectively. N. caninum and T. gondii infection was detected by PCR in 6.8% (5/74) and 5.4% (4/74) of sheep foetuses, respectively, of which five showed protozoal-associated lesions. N. caninum DNA was detected in 11.5% (3/26) of goat foetuses, of which two showed protozoal-associated lesions, whereas T. gondii DNA was detected in one goat foetus with no lesions. The simultaneous presence of N. caninum and T. gondii DNA was detected in one sheep foetus with severe lesions. This study demonstrates that N. caninum plays a significant role in abortion in small ruminants in the studied population. In addition, our results highlight the importance of differentiating between protozoa whenever characteristic lesions are observed.     

Fatal hepatic sarcocystosis in a puppy with eosinophilia and eosinophilic peritoneal effusion

A 3-month-old male Golden Retriever puppy was evaluated for lethargy and fever of 2-days duration. Results of a CBC and biochemical profile revealed marked eosinophilia (6.3 X 10(3)/microL; reference interval 0.1-1.2 X 10(3)/microL), moderate thrombocytopenia, and increased activities of alanine aminotransferase, aspartate aminotransferase, and creatine kinase. Hepatomegaly and peritoneal effusion were found using abdominal ultrasound. Peritoneal fluid analysis revealed eosinophilic inflammation (23,000 nucleated cells/microL with 88% eosinophils). Despite supportive treatment the puppy's condition deteriorated rapidly; euthanasia was requested, and a necropsy performed. Microscopically, there was marked necrosuppurative and eosinophilic hepatitis with vasculitis. Numerous hepatocytes contained protozoal organisms suspected to be Toxoplasma gondii or Neospora caninum. However, serum was negative for both T gondii and N caninum antibodies; polymerase chain reaction assay on hepatic tissue was negative for both organisms; and immunohistochemical evaluation of hepatic tissue using serum raised against T gondii, N caninum, and Sarcocystis neurona also was negative. Schizont morphology suggested that merozoites replicated by endopolygeny, forming rosettes around a central residual body. Transmission electron microscopy revealed that merozoites lacked rhoptries. These findings were consistent with a diagnosis of Sarcocystis canis, an apicomplexan parasite with an unknown life cycle.     

Evaluation of Neospora caninum and Toxoplasma gondii infections in alpaca (Vicugna pacos) and llama (Lama glama) aborted foetuses from Peru

The aim of this study was to investigate the participation of Neospora caninum and Toxoplasma gondii in abortion cases of Peruvian llamas and alpacas. Fifteen aborted foetuses were recovered from two main rearing areas of camelids in Peru (Central or South Andean region). Foetal histopathology was used to detect the presence of protozoal-associated lesions in target organs. N. caninum and T. gondii infections were confirmed by immunohistochemistry (IHC) combined with PCR and by PCR alone, respectively. The influence of the species (llama and alpaca), foetal age (first, second and third gestational periods) and geographical location (Central or South Andean region) of the foetuses was also studied. Thirteen of the samples (26%, 13/50) showed lesions suggestive of protozoal infection. N. caninum infection was detected by either IHC or specific PCR in 14 out of 50 foetuses (28%), of which 8 also showed protozoal-associated lesions. T. gondii DNA was not detected in any of the foetuses analysed. Protozoal infection was more frequent in the foetuses from the second gestational period (P<0.05, Fisher F-test). No significant association was observed between protozoal infection and species or geographical location (P>0.05, chi2 test). The results of the present study indicate that neosporosis should be included during the differential diagnosis of abortion in llamas and alpacas.     

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)     

Serologic prevalence of Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum in horses in Brazil.

OBJECTIVE: To determine serologic prevalence of Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum in horses in Brazil. DESIGN: Prevalence survey. ANIMALS: 101 Thoroughbreds in Brazil. PROCEDURE: Blood samples were obtained from horses and tested for serum antibodies against S neurona by use of an immunoblot procedure with culture-derived S neurona merozoites as antigen, and for serum antibodies against T gondii and N caninum by use of a modified agglutination test with formalin-preserved tachyzoites and mercaptoethanol. RESULTS: Antibodies against S neurona and T gondii were detected in 36 and 16 of 101 horses, respectively. Cross-reactivity between antibodies against T gondii and S neurona was not detected. Antibodies against N caninum were not detected in any samples. CONCLUSIONS AND CLINICAL RELEVANCE: The high prevalence of antibodies against S neurona detected in clinically normal horses emphasizes the importance of examining CSF for antibodies when establishing a diagnosis of equine protozoal myeloencephalitis.     

Neospora-like protozoal infections associated with bovine abortions

Eighty bovine fetuses with presumed protozoal infections from a previous 2-year retrospective study were examined by immunohistochemistry using antisera against Neospora caninum. In 66 (83%) of the fetuses, protozoa were found that reacted positively with anti-N. caninum sera. In three (4%) additional fetuses, protozoa identified as Sarcocystis species did not react, and in two fetuses (3%) single protozoal clusters were found only in hematoxylin and eosin-stained slides. A group of 20 fetuses were chosen for further evaluation. They included 14 fetuses from the first group of 80 fetuses plus six additional fetuses that had large numbers of protozoa in the fetal brain. The 20 fetuses were examined immunohistochemically with antisera to N. caninum, Hammondia hammondi, and Toxoplasma gondii. Protozoa from 3/20 fetuses, identified as Sarcocystis species, failed to react with any antisera. In 16/20 fetuses protozoa reacted positively to antisera against N. caninum, and in most cases reacted to H. hammondi, and weakly to one or more of the antisera against T. gondii. Thick-walled protozoal tissue cysts were found in the brain of four of these 16 fetuses by transmission electron microscopy. The cyst wall morphology was comparable to N. caninum. The results suggested that a single protozoal parasite of unknown identity was responsible for most of the bovine abortions. By immunohistochemistry, the unknown protozoon reacted most strongly and consistently to N. caninum antisera, but was antigenically distinct from N. caninum. Ultrastructurally, tissue cysts found in four fetuses most closely resembled Neospora caninum.     

The role of Neospora caninum and Toxoplasma gondii in spontaneous bovine abortion in Argentina

The aim of the present work was to evaluate the role of Neospora caninum and Toxoplasma gondii infections in spontaneous bovine abortions in Argentina. Based on histopathological results, 70 presumptive cases of apicomplexan protozoal abortion from a total of 666 cases of spontaneous bovine abortion submitted to the National Institute of Agrarian Technology, Balcarce, from 1999 to 2007 were included in this study. N. caninum infection was diagnosed by an indirect fluorescent antibody test (IFAT), by immunohistochemistry (IHC) and by nested-PCR. T. gondii infection also was diagnosed by nested-PCR. DNA from fetuses was extracted primarily from CNS tissues. Heart, liver, muscle and/or placenta were processed when nervous tissue was not available. Sixty-six (9.9%) fetuses were positive by at least one technique (IFAT, IHC or nested-PCR) for N. caninum infection. Overall, there was poor agreement among results obtained by these diagnostic techniques. In contrast, no Toxoplasma-infection was detected in any aborted bovine fetus.     

Seroepidemiology of Neospora caninum and Toxoplasma gondii infection in yaks (Bos grunniens) in Qinghai, China

Neospora caninum is a protozoan parasite and is closely related to Toxoplasma gondii, but they are antigenically different. N. caninum and T. gondii infection in a variety of animals such as cattle, dogs, and cats has been reported, but there is little information on the infection of these parasites in domestic yaks. Seroprevalence of antibodies to T. gondii and N. caninum in yaks (Bos grunniens) from eight regions of Qinghai, China were investigated by the indirect agglutination test (IAT) and ELISA, respectively. A total of 112 (11.8%) of 946 serum samples were positive for antibodies to T. gondii, and 21 samples (2.2%) were positive to N. caninum. Two of the yaks had antibodies to both parasites. There was no apparent association of T. gondii infection with age of the animals. The results indicate that T. gondii infection is prevalent in Chinese yaks in most parts of Qinghai province and N. caninum infection rate in the same species is relatively low. This is the first large study showing the infection of T. gondii and N. caninum in domestic yaks.     

Studies on serological cross-reaction of Neospora caninum with Toxoplasma gondii and Hammondia heydorni

The enzyme-linked immunosorbent assay (ELISA) was used to examine cross-reactivity of Neospora caninum with Toxoplasma gondii and Hammondia heydorni. Anti-T. gondii mouse and cat sera cross-reacted with N. caninum soluble antigen (NLA), but not with the recombinant surface antigen (NcSRS2). Anti-H. heydorni dog sera showed no cross-reactivity with either the NLA antigen or the NcSRS2. Lack of cross-reactivity between anti-H. heydorni sera and N. caninum antigens, and the cross-reactivity of anti-T. gondii sera with the NLA suggest that N. caninum has common antigens to T. gondii except for NcSRS2 based on serology. In light of several studies suggesting a closer relationship between N. caninum and H. heydorni than with T gondii, examination of serological cross-reactivity with N. caninum may be necessary to further classify the parasites in addition to molecular and morphological studies and clarification of the life cycle.     

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).     

Detection of Toxoplasma gondii-like oocysts in cat feces and estimates of the environmental oocyst burden

OBJECTIVE: To estimate the analytic sensitivity of microscopic detection of Toxoplasma gondii oocysts and the environmental loading of T gondii oocysts on the basis of prevalence of shedding by owned and unowned cats. DESIGN: Cross-sectional survey. SAMPLE POPULATION: 326 fecal samples from cats. PROCEDURES: Fecal samples were collected from cat shelters, veterinary clinics, cat-owning households, and outdoor locations and tested via ZnSO(4) fecal flotation. RESULTS: Only 3 (0.9%) samples of feces from 326 cats in the Morro Bay area of California contained T gondii-like oocysts. On the basis of the estimated tonnage of cat feces deposited outdoors in this area, the annual burden in the environment was estimated to be 94 to 4,671 oocysts/m(2) (9 to 434 oocysts/ft(2)). CONCLUSIONS AND CLINICAL RELEVANCE: Despite the low prevalence and short duration of T gondii oocyst shedding by cats detected in the present and former surveys, the sheer numbers of oocysts shed by cats during initial infection could lead to substantial environmental contamination. Veterinarians may wish to make cat owners aware of the potential threats to human and wildlife health posed by cats permitted to defecate outdoors.     

Aetiology of bovine abortion in Switzerland from 1986 to 1995--a retrospective study with emphasis on detection of Neospora caninum and Toxoplasma gondii by PCR

In a retrospective study, covering the period from 1986 to 1995, tissues of aborted fetuses were re-examined. A total of 347 cases were tested immunohistochemically, among them samples of 223 brains were examined for Neospora caninum, Toxoplasma gondii and Bovine Virus Diarrhoea Virus (BVDV), and 249 placentae for Chlamydiaceae. Two real-time PCR assays, one for N. caninum, and one for T. gondii, were developed. These potential abortion-inducing agents were detected - and confirmed by PCR, except for BVDV - in 16.1% (N. caninum), 0% (T. gondii), 9.9% (BVDV) and 0.8% (Chlamydiales) of the cases examined. Immunohistochemistry proved to be inadequate for the detection of the protozoal epitopes, whereas it was confirmed as a very useful tool for the detection of BVDV. In abortion material, PCR is considered to be more suitable for the detection of protozoa and Chlamydophila abortus, an adequate sampling presupposed.     

Seroepidemiology of Neospora caninum and Toxoplasma gondii in cattle and water buffaloes (Bubalus bubalis) in the People's Republic of China

A seroepidemiological survey of Neospora caninum and Toxoplasma gondii in cattle and water buffaloes was carried out in the People's Republic of China. Serum samples were obtained from dairy (n=262, 9 herds in 9 provinces) and beef cattle (n=10, 1 herd) and water buffaloes (n=40) in China. All sera were tested for antibodies to N. caninum and T. gondii by an enzyme-linked immunosorbent assay (ELISA) and an indirect agglutination test (IAT), respectively. The overall seroprevalence of N. caninum in dairy cattle was 17.2% (45/262), and the herds seroprevalence of N. caninum was 88.9% (8/9), and antibodies to T. gondii were present in 6 cows (2.3%). None of the cows had antibodies against both T. gondii and N. caninum. Antibodies to T. gondii or N. caninum were not found in beef cattle or water buffaloes. The seroprevalence of N. caninum in aborting cows (20.2%) was higher than that in non-aborting cows (16.6%) with an odds ratio of 1.26 (95% CI, 0.54-2.95), but the difference was not statistically significant (P>0.05). There was no apparent association of N. caninum seropositivity with age or number of pregnancies. This is the first report on the seroprevalence of N. caninum in cattle and water buffaloes in China.     

Abortions caused by protozoa in agricultural animals]

A review is given on abortions in livestock caused by Toxoplasma gondii, Neospora caninum and Sarcocystis spp. Special emphasis is put on diagnostic procedures. T. gondii is considered to be a major cause of abortions in sheep and goats. Diagnosis is based on the characteristic lesions in the cotyledons, histologic and immunohistochemical examination, and on the detection of specific antibodies in body fluids or serum of aborted fetuses. Whether abortions in swine due to T. gondii are of practical importance is still unclear. The most important causative protozoal agent of abortions in cattle is Neospora caninum. This protozoon was found in 19 per cent of cattle fetuses submitted for examination in USA. Occasionally it was found also in aborted lambs, kids and foals. Diagnosis is based on the histologic or immunohistochemical detection of the parasite. Abortions can be regularly induced experimentally in large and small ruminants and in pigs by the oral inoculation of sporocysts of certain Sarcocystis species. However, under natural conditions abortion due to Sarcocystis was diagnosed only occasionally and only in cattle and sheep. The histologic examination of the fetus and fetal membranes is the only way to diagnose Sarcocystis abortion or neonatal infection at present.     

Seroprevalence of Toxoplasma gondii and Neospora caninum in dairy goats from Romania

Little information is available about the seroprevalence of Toxoplasma gondii and Neospora caninum infections in goats in Romania and even in Europe. During 2007-2010, 735 serum samples were collected from dairy goats located in 4 historical regions (Cri?ana, Maramure?, Transylvania and Muntenia) of Romania. Sera were analyzed for T. gondii and N. caninum antibodies (IgG type) by enzyme-linked immunosorbent assay (ELISA) using two commercial kits (Chekit Toxotest Antibody ELISA and Chekit Neospora caninum Antibody ELISA; Idexx-Bommeli, Switzerland). Three hundred and eighty-eight out of 735 (52.8%) goats presented T. gondii antibodies and 12 out of 512 (2.3%) goats had N. caninum antibodies. The high seroprevalence of T. gondii suggests that infection with this parasite is common in dairy goats in Romania, and less common the infection with N. caninum. This is the first time that infection with N. caninum in goats has been reported in Romania and the first extended study on seroepidemiology of T. gondii.