Sarcocystis Spp Research Articles

Oropharyngolaryngeal Sarcocystosis and Histoplasmosis Co-Infection in a Patient with Systemic Lupus Erythematosus

Sarcocystosis is an uncommon zoonotic coccidian protozoal infection caused by Sarcocystis spp. Muscular sarcocystosis have been detected in patients with various types of cancer but co-infection with histoplasmosis has never been reported. We report a 39-year-old woman with active lupus nephritis who presented with hoarseness for eleven months. Physical examination and magnetic resonance imaging revealed an irregular exophytic mass at the left oropharynx and the left true vocal cord. The histopathologic diagnosis was oropharyngolaryngeal sarcocystosis and histoplasmosis. She was treated with oral itraconazole for histoplasmosis. The authors briefly review the clinicopathologic features and pathogenesis of oropharyngolaryngeal sarcocystosis and histoplasmosis and conclude the possibility of co-infection of oropharyngolaryngeal sarcocystosis and histoplasmosis, especially in immunocompromised patient.

Detection of Toxoplasma gondii and Sarcocystis tenella in indigenous Cornigliese sheep in Italy using serological and molecular methods

The aim of the present study was to determine seroprevalence for Toxoplasma gondii by meat juice ELISA and evaluate the presence of T. gondii and Sarcocystis spp. within host tissues by histology, PCR and in vitro isolation, in the indigenous Cornigliese sheep breed in northern Italy. Seventeen out of 24 (70.8%) sheeps were positive for T. gondii by meat juice ELISA. Twenty sheep (83.3%) were positive by PCR for T. gondii, while 24/24 sheep (100%) were positive by PCR for Sarcocystis spp. Tissues cysts compatible with Sarcocystis spp. were visible in all animals on histology. PCR confirmed the presence of T. gondii after three weeks of in vitro culture on Vero cells in only one sample. Genotyping of T. gondii by RLFP with 5 markers showed a predominance of genotypes II/III. Sequence analysis of Sarcocystis spp. showed only the presence of Sarcocystis tenella.T. gondii and S. tenella are present in a high percentage of Cornigliese sheep in northern Italy. Future studies should concentrate upon the reproductive and economic effects of these parasitic infections, in light of the necessary conservation of this local, indigenous sheep breed.

Sarcocystis spp. Infection in two Red Panda Cubs (Ailurus fulgens)

Two neonatal male red panda (Ailurus fulgens) littermates were submitted for necropsy examination. One animal was found dead with no prior signs of illness; the other had a brief history of laboured breathing. Post-mortem examination revealed disseminated protozoal infection. To further characterize the causative agent, transmission electron microscopy (TEM), immunohistochemistry (IHC), polymerase chain reaction (PCR) and amplification and nucleic acid sequencing were performed. IHC was negative for Toxoplasma gondii and Neospora caninum, but was positive for a Sarcocystis spp. TEM of cardiac muscle and lung revealed numerous intracellular apicomplexan protozoa within parasitophorous vacuoles. PCR and nucleic acid sequencing of partial 18S rRNA and the internal transcribed spacer (ITS)-1 region confirmed a Sarcocystis spp. that shared 99% sequence homology to Sarcocystis neurona and Sarcocystis dasypi. This represents the first report of sarcocystosis in red pandas. The histopathological, immunohistochemical, molecular and ultrastructural findings are supportive of vertical transmission resulting in fatal disseminated disease.

32 ACTIVATION OF REDOX-SENSITIVE INFLAMMASOMES UNDERLIES THE BIOLOGICAL PHENOTYPE OF MYELODYSPLASTIC SYNDROMES

The aim of the present study was to determine seroprevalence for Toxoplasma gondii by meat juice ELISA and evaluate the presence of T. gondii and Sarcocystis spp. within host tissues by histology, PCR and in vitro isolation, in the indigenous Cornigliese sheep breed in northern Italy. Seventeen out of 24 (70.8%) sheeps were positive for T. gondii by meat juice ELISA. Twenty sheep (83.3%) were positive by PCR for T. gondii, while 24/24 sheep (100%) were positive by PCR for Sarcocystis spp. Tissues cysts compatible with Sarcocystis spp. were visible in all animals on histology. PCR confirmed the presence of T. gondii after three weeks of in vitro culture on Vero cells in only one sample. Genotyping of T. gondii by RLFP with 5 markers showed a predominance of genotypes II/III. Sequence analysis of Sarcocystis spp. showed only the presence of Sarcocystis tenella.T. gondii and S. tenella are present in a high percentage of Cornigliese sheep in northern Italy. Future studies should concentrate upon the reproductive and economic effects of these parasitic infections, in light of the necessary conservation of this local, indigenous sheep breed.

PREVALENCE OF BRAIN LESIONS IN SHEEP SLAUGHTERED AT MOSUL AREA DURING 2012-2013.

The present study revealed the prevalence and pathology of sheep brain(male and female). One handred brain samples were obtained randomly from Mosul animal slaughter house and butcher shops since October 2012 to August 2013 from animals with and without nervous signs. The main gross lesions were edema which accounted 53% of the cases, congestion 51% , hemorrhage 20% and 3% cyst of coenurous cerebralis. The major lesions were observed by histopathological examination of three different areas of the brain (cerebrum, cerebellum and medulla oblongata)in each case. Lesions include vascular change, neuronal degeneration, necrosis, inflammatory change, gliosis, neuronophagia, amyloid deposition. Three cases revealed the presence of the coenurous cerebralis and two cases were abscesses and one case showed the presence of Sarcocystis SPP.

Sarcocystis sinensis is the most prevalent thick-walled Sarcocystis species in beef on sale for consumers in Germany

Bovines are intermediate hosts of Sarcocystis cruzi, Sarcocystis hirsuta, and Sarcocystis hominis, which use canids, felids, or primates as definitive hosts, respectively. Cattle represent also intermediate hosts of Sarcocystis sinensis, but the definitive hosts of this parasite are not yet known. Sarcocystosis in cattle is frequently asymptomatic. The infection is characterized by the presence of thin-walled (S. cruzi) or thick-walled muscle cysts or sarcocysts (S. hominis, S. sinensis, and S. hirsuta). Recent reports suggest high prevalence of the zoonotic S. hominis in beef in Europe. We therefore aimed at differentiating Sarcocystis spp. in beef offered to consumers in Germany using molecular and microscopical methods, focusing on those species producing thick-walled sarcocysts. A total of 257 beef samples were obtained from different butcheries and supermarkets in Germany and processed by conventional and multiplex real-time PCR. In addition, 130 of these samples were processed by light microscopy and in 24.6% thick-walled cysts were detected. Transmission electron microscopical analysis of six of these samples revealed an ultrastructural cyst wall pattern compatible with S. sinensis in five samples and with S. hominis in one sample. PCR-amplified 18S ribosomal DNA (rDNA) fragments of 28 individual thick-walled cysts were sequenced, and sequence identities of ≥98% with S. sinensis (n = 22), S. hominis (n = 5) and S. hirsuta (n = 1) were observed. Moreover, nine Sarcocystis sp. 18S rDNA full length gene sequences were obtained, five of S. sinensis, three of S. hominis, and one of S. hirsuta. Out of all samples (n = 257), 174 (67.7%) tested positive by conventional PCR and 179 (69.6%) by multiplex real-time PCR for Sarcocystis spp. Regarding individual species, 134 (52%), 95 (37%), 17 (6.6%), and 16 (6.2%) were positive for S. cruzi, S. sinensis, S. hirsuta, and S. hominis, respectively. In conclusion, S. sinensis is the most prevalent thick-walled Sarcocystis species in beef offered for consumption in Germany. Further studies are needed to identify the final host of S. sinensis as well as the potential role of this protozoan as a differential diagnosis to the zoonotic species S. hominis.

Phylogenetic analysis and identification of Sarcocystis spp. found in rodents in Peninsular Malaysia

Methods: Histological sections of these rodent muscle tissues stained with hematoxylin and eosin showed infection with Sarcocystis spp. Based on these results, the current study was carried out to determine the phylogenetic relationship among the identified Sarcocystis spp. in these rodents. The formalin fixed paraffin embedded (FFPE) rodent muscle blocks were subjected to DNA extraction and followed with semi nested PCR targeting 5’ and 3’ regions of 18S rRNA of Sarcocystis spp.

Sarcocystis spp. in llamas (Lama glama) in Southern Bolivia: A cross sectional study of the prevalence, risk factors and loss in income caused by carcass downgrades

Llamas (Lama glama) are intermediate hosts of the protozoan parasite Sarcocystis spp. This parasite is described as causing economic losses in the production of llama meat in South America. The aim of this study was to estimate prevalence, identify risk factors and explore spatial patterns of Sarcocystis in llamas in an area of the Bolivian High Plateau including estimating financial losses due to carcass downgrades as a result of the presence of Sarcocystis cysts.Information was collected from a local abattoir between 2006 and 2011 on 1196 llamas. Sarcocystis status was determined at meat inspection where any carcasses with one or more visible cysts were deemed Sarcocystis positive. A high prevalence was found, estimated to vary between 23.4% (95% CI 16.6–30.1) in 2007 and 50.3% (95% CI 44.4–56.3) in 2011. Period prevalence between 2006 and 2011 was estimated at 34.1% (95% CI 31.4–36.8). Age, sex and type (analogous to breed) were identified as risk factors for Sarcocystis using a mixed-effects logistic regression model adjusting for clustering by community and owner. Llamas over 4.5 years of age had an increased odds of being Sarcocystis positive (OR 19.31, 95% CI 9.10–40.98) as well as females (OR 1.75, 95% CI 1.13–2.68) and long haired type llamas (OR 1.90, 95% CI 1.26–2.87). An interaction between age and sex was detected indicating that the increased odds of disease from the youngest age group to the 2.5–4.5 years group was much more pronounced in females than in males. Spatial patterns of Sarcocystis were explored at district level by means of Standardised Morbidity Ratios and some spatial heterogeneity was revealed. Estimates of financial loss due to the disease were calculated using the difference in price paid for Sarcocystis positive and negative meat. Loss due to Sarcocystis varied per year but could be up to 20% of the annual income generated through the abattoir by sale of meat.Overall this study shows a high prevalence of Sarcocystis in the study area with some heterogeneity between districts. It also identifies some previously unknown risk factors for Sarcocystis and gives financial estimates of the cost of the disease as a result of carcass downgrades. We hoped these findings will add to the understanding of Sarcocystis in llamas in Southern Bolivia and will be useful when considering if controls are necessary, worthwhile and practical.

PREVALENCE OF SARCOCYSTIS SPP. IN SHEEP AND GOATS AND ITS EFFECT ON SOME BLOOD CONSTITUENTS IN SHARKIA PROVINCE

This study was carried out on slaughtered sheep and goats in different slaughter houses in Sharkia Province to determine the prevalence of ovine and caprine Sarcocystis infection and effect on some biochemical clinical pathological parameters. Parasitological examination of faecal samples revealed presence of eggs of internal parasites in 25(41.67%) lambs(less 9 month) {12(48%) nematodes, 7(28%) trematode ) & 6(24%) mixed infection} 29 (48.33%) adult (more 2 year) ( sheep {15(51.72%) nematodes,10 (34.48%) trematode & 4(13.79%) mixed infection},19 (47.50%) young goats {11(57.89%) nematodes, 5(26.32%) trematode & 3(15.79%) mixed infection}, 24 (60%) adult goats{14(58.33%) nematodes, 5(20.83%) trematode &5 (20.83 %) mixed infection}. Serologically, antibodies against Sporocystis spp. infection were observed in sheep serum samples were 47.50% (57) & 40% (32) for goats serum samples. The infection in adult animals were {34 (56.67%) in sheep and 19 (47.50%) in goats} more than in young animals {23 (38.33%) in sheep and13 (32.50 % in goats}. Sarcocystis in sheep distributed as macrocystis (16.67% for young and 11.67% adult), microcystis ovi-cans (10% young, 5% adult) and mixed infection (macrocystis and microcystis) were (56.67% young, 38.33% adult), meanwhile in goats Sarcocystis macrocystis (25% young and 15% adult), microcystis (capricans) (15% young,12.5% adult) and mixed infection (macrocystis and microcystis) 47.50% for young, 32.50%for adult). Sarcocystis infection in Sheep and goats induced a significant decrease in leukocytic count, monocytes, total protein, albumin, globulin, lymphocytosis, neutrophilia, non significant effect in basophils and eosinophilis beside significant rise AST,ALT& ALP. It could be concluded that the Sarcocystis spp infected both sheep and goat, the infection in adult were more than in young ones. Sarcocystis induces changes in biochemical parameters. Therefore, it has of great importance the human to be trained not to feed their dogs and cats with uncooked meat, and abattoir remnants were burned to be effectively broken of infection cycle between the intermediate and the definitive hosts.

Molecular identification of Sarcocystis aucheniae as the macrocyst-forming parasite of llamas

The domestic South American camelids (SACs), llama (Lama glama) and alpaca (Lama paco), are frequently found to be infected with Sarcocystis parasites. Infections give rise in skeletal muscle to macroscopic cysts (1–5mm long) that resemble rice seeds, each containing several million living bradyzoites. The finding of cysts prevents commercialization of SAC meat, an important source of income for rural families in the Andean flatlands. Thus, development of diagnostic methods to facilitate the control of these infections is highly desirable, and the first step to this end is the unequivocal species identification of the causative agent. Based on the cyst form and size, the infecting parasite has been described as Sarcocystis aucheniae; however, this traditional approach is not reliable as similar cysts may contain different species. To date, molecular identification has been done for a single isolate of S. aucheniae from an alpaca in Australia. In order to verify the identity of the species present in SACs of South America, the complete 18S rRNA gene was PCR-amplified and sequenced from macrocyst DNA obtained from three llamas of the Andean flatlands. A phylogenetic Bayesian analysis was carried out using the analyzed and available 18S rRNA sequences of Sarcocystis spp. In the constructed tree, all of the new 18S rRNA gene sequences segregated in a single clade together with the 18S rRNA gene sequence reported from an alpaca in Australia, demonstrating that the isolated parasite is S. aucheniae, and that this parasite indiscriminately infects both domestic SACs. This work represents the first molecular identification of the causative agent of SAC sarcocystiosis in South America, and can contribute to the development of control methods for this neglected parasitosis.

Phylogenetic relationships among Sarcocystis species in cervids, cattle and sheep inferred from the mitochondrial cytochrome c oxidase subunit I gene

Coccidian parasites in the genus Sarcocystis have a two-host life cycle, and have traditionally been identified on the basis of morphological features of the sarcocyst stage in their intermediate hosts. Additional molecular species identification, delimitation and phylogeny of Sarcocystis spp. have been based mainly on the nuclear ssrRNA gene. This gene is well suited for discrimination between more distant species but less so for closely related species. The objective of this study was therefore to establish the mitochondrial cytochrome c oxidase subunit I gene (cox1) as a novel genetic marker for Sarcocystis spp. and assess its utility for species identification and delimitation. New primers were developed and 1,020–1,095bp long cox1 sequences were obtained from 155 isolates of 22 Sarcocystis spp. from cattle, sheep, red deer, reindeer, roe deer and moose, and used for phylogenetic reconstructions. For 18 species, the intraspecific and interspecific sequence identities were 98.5–100% and 58–92%, respectively. The four other species had previously been regarded as two species (Sarcocystis rangiferi, Sarcocystis tarandi), each infecting both reindeer and red deer. From cox1 data, each of those appeared to be two separate species, with S. rangiferi and S. tarandi being restricted to reindeer. Thus, cox1 sequences seem to perform better than ssrRNA gene sequences for delimitation of closely related species. The 22 species were distributed in three major clades according to their definitive hosts as in phylogenetic trees obtained from the ssrRNA gene. There were only minor differences in the branching order of different taxa between the trees obtained from either gene. This study has successfully established cox1 as a novel genetic marker for future research on Sarcocystis spp. It has also provided the first published molecular identification of Sarcocystis gigantea and Sarcocystis tenella in Norwegian sheep, and of Sarcocystis hirsuta and Sarcocystis sinensis in Argentinean cattle.

Infections with Sarcocystis wenzeli are prevalent in the chickens of Yunnan Province, China, but not in the flocks of domesticated pigeons or ducks

The distribution and prevalence of infections with species of Sarcocystis in domestic fowl in Asia are poorly known. Here, ducks, pigeons, and chickens from Yunnan Province, China were examined for evidence of parasitic infection with Sarcocystis spp. One hundred and ninety one chickens, 514 ducks, and nine pigeons were investigated. Whereas the ducks and pigeons lacked tissue cysts in their muscle, brain or peripheral nervous system, cysts of Sarcocystis wenzeli were identified in 17 of 191 chickens (8.9%). Morphologically, the cysts were thread-like, ranging in size from 334–3169×41–117μm (mean 1093×65μm). Cysts were septate with dense, short finger-like protrusions which appeared radially striated. The cyst wall was 1.4–3.5μm (mean 2.4μm) thick. The bradyzoites were lancet shaped and measured 12.2–17.7×1.8–2.9μm (mean 14.6×2.5μm). Ultrastucturally, the primary sarcocyst wall had stubby villar protrusions, corresponding to the ‘type 9’ class previously designated. The protrusions measured 0.87–1.89×0.47–0.91μm (mean 1.27×0.59μm; n=57). These findings confirm previous work from the vicinity of Kunming concerning the occurrence of S. wenzeli in chickens, and its use of both cats and dogs as definitive hosts, but indicate that corresponding infections may not occur in the regional domestic flocks of other types of fowl.

FRECUENCIA DE sarcocystis sp. EN PERROS PASTORES DE ASOCIACIONES ALPAQUERAS DE MARANGANÍ, CUSCO.

The aim of the study was to determine the frequency of Sarcocystis spp. in sheepdogs of alpaca farm associations at Maranganí, Cusco, during the dry and rainy season (April 2005 until February 2006). Fecal samples (n = 211) were collected and evaluated using the method of flotation with Sheather solution to observe sporocysts of Sarcocystis spp. The frequency during the dry and rainy season was 42.3 ± 9.2% (47/111) and 72.0 ± 8.8% (72/100) respectively. The results showed an association between the rate of infection and the time of the year (p<0.05), where November had the highest rate (77.8 ± 12.1%, 35/ 45) and March had the lowest (17.6 ± 12.8%, 6/34). In addition, significant difference was found due to age of the dog (p<0.05).

Exposure to Sarcocystis spp. in horses from Spain determined by Western blot analysis using Sarcocystis neurona merozoites as heterologous antigen

Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs.

Fatal Hepatic Sarcocystosis in a Captive Black Bear (Ursus Americanus) Associated with Sarcocystis Canis-Like Infection

Fatal hepatic sarcocystosis was diagnosed in a 13-year-old captive black bear (Ursus americanus) with a history of acute onset of vomiting, polyuria, polydipsia, and bilirubinuria. Gross lesions included severe icterus, multisystemic hemorrhage, and gall bladder edema. The most significant microscopic lesion was severe necrotizing hepatitis with intralesional protozoa that reproduced by endopolygeny consistent with a Sarcocystis spp. Infrequent microglial nodules were randomly scattered within the white matter of the cerebral cortices, thalamus, and brainstem, but intralesional protozoal schizonts were not observed. In the liver, immunohistochemistry was positive for Sarcocystis spp. and negative for Toxoplasma gondii and Neospora spp. Positive staining was not observed in the brain. Genus-specific polymerase chain reaction (PCR) amplification of the 18S ribosomal RNA gene was performed on formalin-fixed, paraffin-embedded sections of liver and brain; in both tissues, PCR was positive for Sarcocystis spp. Sequence analysis of the PCR amplicons revealed 100% identity to the published sequences of Sarcocystis canis and Sarcocystis arctosi.

Prevalence to Toxoplasma gondii and Sarcocystis spp. in a Reintroduced Fisher (Martes pennanti) Population in Pennsylvania

Understanding the role of disease in population regulation is important to the conservation of wildlife. We evaluated the prevalence of Toxoplasma gondii exposure and Sarcocystis spp. infection in 46 road-killed and accidentally trapper-killed fisher (Martes pennanti) carcasses collected and stored at -20 C by the Pennsylvania Game Commission from February 2002 to October 2008. Blood samples were assayed for T. gondii antibodies using the modified agglutination test (MAT, 1 : 25) and an indirect immunofluorescent antibody test (IFAT, 1 : 128). For genetic analysis, DNA samples were extracted from thoracic and pelvic limb skeletal muscle from each carcass to test for Sarcocystis spp. using 18s-rRNA PCR primers. Antibodies to T. gondii were found in 100% (38 of 38) of the fishers tested by MAT and in 71% (32 of 45) of the fishers tested by IFAT. PCR analysis revealed that 83% (38 of 46) of the fishers were positive for Sarcocystis spp. Sequence analysis of 7 randomly chosen amplicons revealed the fisher sarcocysts had a 98.3% to 99.1% identity to several avian Sarcocystis spp. sequences in GenBank. Data from our study suggest that a high percentage of fishers in Pennsylvania have been exposed to T. gondii and are infected with Sarcocystis spp.

Risk Assesment Regarding the Sarcocystis spp. Infestation at Ovine Carcasses Destined for Public Consumption

According to the bibliographical studies' analysi s, the incidence of ovine sarcocystosis, is situated in the first place reach ing values of over 98%. The field researches concern the intensity and extent degree of the micr osarcocysts in the skeletal muscles, in the following anatomic regions: the intercostal muscle s, the diaphragm muscle, the tongue muscles, the dorsal muscles and the thigh muscles, with aspects regarding the incidence of these microcysts, depending on the animals' age. Th ere were 160 meat samples collected from 20 ovine aged between 1,5 and 4 years. Following the samples' processing there was noticed that the extent of the Sarcocystis spp. infestation at the samples evaluated was of 100%, the average value of the intensity being the highest in the myocardia. In this context, it is necessary, in the case of ovine meat products, w hich do not involve in their technological process a thermic treatment, to priory make a contr ol also to investigate the microcysts evaluation. At the same time, the ovine meat and or gans administrated in the carnivore's diet must be thermic sterilized.

Detection of sarcocystic infection in a wild rodent (Apodemus agrarius chejuensis) captured on Jeju island

Sarcocystis spp is a causative agent of sarcocystosis. They have a characteristic life cycle infecting both prey and predator. Sarcocystis can cause myositis, atrophy of the adjacent cells and abortion in cattle. In mice, sarcocystosis causes mild cellular reactions without clinical disease. Severe haemorrhage and abortion were also reported. For monitoring the disease in wild rodents of the Korean peninsula, we captured Apodemus agrarius chejuensis on Jeju island and examined the specimen histopathologically. Intramuscular cysts were found and diagnosed as Sarcocystis. Sarcocystic infection has been reported in worldwide. There have been many reported infections in cattle and pigs in Korea. To our knowledge, this is the first report of Sarcocystis in Apodemus agrarius chejuensis captured in Korea.

High prevalence of Sarcocystis calchasi sporocysts in European Accipiter hawks

The emerging Sarcocystis calchasi induces a severe and lethal central nervous disease in its intermediate host, the domestic pigeon (Columba livia f. domestica). Experimental studies have identified the Northern goshawk (Accipiter g. gentilis) as final host. Phylogenetically closely related European sparrowhawks (Accipiter n. nisus) and wood pigeons (Columba palumbus) have been found to harbor genetically closely related Sarcocystis spp. However, data on the prevalence and potential interspecies occurrence of these parasites are lacking. Here, we report that European Accipiter hawks (Accipitrinae) are highly infected with S. calchasi, S. columbae and Sarcocystis sp. ex A. nisus in their small intestine. Thirty-one of 50 (62%) Northern goshawks necropsied during 1997–2008 were positive for S. calchasi in a newly established species-specific semi-nested PCR assay based on the first internal transcribed spacer region. Unexpectedly, 14 of 20 (71.4%) European sparrowhawks tested also positive. In addition, birds of both species were found to be infested with S. columbae and an, as yet, unnamed Sarcocystis sp. recently isolated from European sparrowhawks. These findings raise new questions about the host specificity of S. calchasi and its high virulence in domestic pigeons, since sparrowhawks only rarely prey on pigeons. Notably, isolated sporocysts from both infected Accipiter spp. measured 8μm×11.9μm, precluding a preliminary identification of S. calchasi in feces of Accipiter hawks based on morphology alone. Importantly, three of four Northern goshawks used in falconry tested positive for S. calchasi. In conclusion, the results indicate that both European Accipter spp. in Germany serve as natural final hosts of S. calchasi and suggest that falconry and pigeon sport may serve as risk factors for the spread of this pathogen in domestic pigeons.

Meningoencephalitis Associated with Sarcocystis spp. in a Free-Living Japanese Raccoon Dog ( Nyctereutes procyonoides viverrinus)

A free-living, young adult, male Japanese raccoon dog (Nyctereutes procyonoides viverrinus) was rescued in Gifu, Japan in March 2009. The animal was weak and emaciated with neurological signs that included head tilt, tremor and tic. The brain showed no gross abnormality at necropsy, but microscopically there was severe meningoencephalitis associated with protozoa, which were morphologically consistent with the asexual developmental stage of Sarcocystis spp. The protozoa were immunohistochemically negative for Toxoplasma gondii and Neospora caninum, but reacted weakly with antiserum specific for Sarcocystis cruzi. Analysis of the partial 18S rRNA gene sequence revealed that the protozoa were most closely related to an unidentified Sarcocystis species that was isolated from the white-fronted goose (Anser albifrons).