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Sarcocystis neurona, a protozoal parasite shed by opossums (Didelphis virginiana), has been shown to cause significant morbidity and mortality in horses, sea otters, and other marine mammals. Over the course of 3 years (fall 2005-summer 2008), opossums from central California were tested for infection with S. neurona. Of 288 opossums sampled, 17 (5.9%) were infected with S. neurona based on the molecular characterization of sporocysts from intestinal scrapings or feces. Risk factors evaluated for association with S. neurona infection in opossums included: age, sex, location, season, presence of pouch young in females, concomitant infection, and sampling method (live-trapped or traffic-killed). Multivariate logistic regression analysis identified that opossums in the Central Valley were 9 times more likely to be infected than those near the coast (p=0.009). Similarly, opossum infection was 5 times more likely to be detected during the reproductive season (March-July; p=0.013). This first investigation of S. neurona infection prevalence and associated risk factors in opossums in the western United States can be used to develop management strategies aimed at reducing the incidence of S. neurona infections in susceptible hosts, including horses and threatened California sea otters (Enhydra lutris neries).

The parasite of genus Sarcocystis is one of the most commonly found parasite in domestic animals worldwide. Some species of Sarcocystis cause important economic loss when causing clinical and sub clinical disease. The aim of this study was to determine the prevalence of Sarcocystis in slaughtered Cattle in Kerman, Iran. The prevalence of Sarcocystis spp. infection was investigated in 480 cattle, slaughtered from May 2005 to February 2006 in the Kerman, Iran using naked eye examination for macroscopic Sarcocysts, and peptic digestion, muscle squash, squeezing methods for microscopic types. Muscles from heart, tongue, and esophagus, cervical and abdominal muscles of 480 slaughtered cattle were examined for Sarcocystis cysts. The prevalence of microscopic Sarcocystis cysts in cattle was detected in 100% and there was no macroscopic cyst in examined cattle.

A total of 65 free-living carnivores collected on Honshu Island, Japan were examined for muscular Sarcocystis species infections. Among them, 12 Japanese raccoon dogs (Nyctereutes procyonoides viverrinus), one Japanese red fox (Vulpes vulpes japonica), three Japanese martens (Martes melampus melampus), and two Japanese badgers (Meles meles anakuma) were found to have sarcocysts in their muscles. No inflammatory reactions associated with sarcocysts were observed. Ultrastructurally, the sarcocysts detected in the Japanese raccoon dogs, Japanese red fox, and Japanese martens were similar to each other, with the sarcocyst wall being thin and exhibiting minute undulations. On the other hand, the sarcocysts detected in the Japanese badgers had a thick cyst wall with numerous finger-like protrusions which contained microtubules. The species of Sarcocystis in Japanese carnivores remain to be determined. This is the first published report on muscular sarcocystosis in Japanese carnivores.

The present study surveyed the prevalence of natural infection of the sheep esophagus muscle with sarcocysts of Sarcocystis ovicanis and examined induction of protective immunity using UV-attenuated sporocysts. The overall prevalence of natural infection of the sheep was 95%. Infectivity of the collected sarcocysts was confirmed by shedding of sporulated oocysts after feeding infected esophageal tissues to dogs. To induce protective immunity, lambs were immunized 3 times (once a week) with 1.5 x 10(4) sporocysts exposed to UV-light for 30 min (UV-30 group) or 60 (UV-60 group) min and then challenged with 1.5 x 10(4) normal sporocysts at the 3rd week post the 1st vaccination. These lambs showed high survival and less clinical signs of sarcocystosis than normal infected lambs. The attenuated sporocysts produced abnormal cysts; small in size and detached from the muscle fiber. These abnormalities were more obvious in UV-60 group than UV-30 group. Also, the IFN-gamma level and lymphocyte percentage were increased while the total leukocyte count was decreased in the UV-60 group compared with other groups. The high level of IFN-gamma may be an evidence for the induction of Th1 responses which may have protective effect against a challenge infection.

Thirty-eight captive psittacine birds housed in a bird park in Foz do Iguaçu, Parana, Brazil, died within a 15-month period as a result of infection with Sarcocystis falcatula. Although fatalities affected 16 species of psittacine birds, mortality was highest in Old World species, which were most susceptible to the pulmonary form of sarcocystosis. Along with the pathologic findings of this disease outbreak, a review of the pathophysiology of sarcosporidiosis is presented.

During 2001 to 2004, 4 species of hawks (Buteo and Accipiter spp.) from Georgia were surveyed for Sarcocystis spp. infections by examining intestinal sections. In total, 159 of 238 (66.8%) hawks examined were infected with Sarcocystis spp. Samples from 10 birds were characterized by sequence analysis of a portion of the 18S rRNA gene (783 base pairs). Only 3 of the 10 sequences from the hawks were identical; the remainder differed by at least 1 nucleotide. Phylogenetic analysis failed to resolve the position of the hawk Sarcocystis species, but they were closely related several Sarcocystis species from raptors, rodents, and Sarcocystis neurona. The high genetic diversity of Sarcocystis suggests that more than 1 species infects these 4 hawk species; however, additional molecular or experimental work will be required to determine the speciation and diversity of parasites infecting these avian hosts. In addition to assisting with determining species richness of Sarcocystis in raptors, molecular analysis should be useful in the identification of potential intermediate hosts.

The prevalence of Sarcocystis spp. was investigated by gross and histopathological examinations in 250 camels (Camelus dromedarius) slaughtered from 2002 to 2005 in the Mashhad Slaughterhouse, eastern Iran. Samples were taken from the diaphragm, heart, tongue, esophagus and masseter muscles for histopathological studies. No macroscopic sarcocysts were found in the samples at gross inspection. Sarcocysts were detected in 209 of 250 (83.6%) examined camels at histopathological level. The infection rate of the esophagus, heart, masseter muscles, diaphragm, and tongue was 58.8%, 48.0%, 46.8%, 41.6%, and 28.0%, respectively. There was no significant difference in the rate of infection between male (85.8%) and female (81.0%) camels. The tissue response to vital cysts was minimal; however, reaction to the degenerating cysts was severe and caused tissue damages resulting in hyperemia, hemorrhages, mononuclear cell infiltration, necrotic changes, and fibrosis. The wild and domestic carnivores especially dogs may be the final hosts of Sarcocystis spp. in this area.

This report describes an outbreak of acute pulmonary sarcocystosis in different species of captive psittacines and in a Luzon bleeding-heart pigeon (Gallicolumba luzonica) in a zoological collection in Brazil. A majority of the birds were found dead and had exhibited no previous clinical signs. Grossly, pulmonary congestion and edema were the most-common findings. Enlarged and congested livers and spleens were also frequently observed. Microscopically, there was edema, fibrin exudation, congestion, and perivascular and interstitial lymphoplasmacytic infiltration associated with numerous sinuous schizonts of Sarcocystis sp. in the lungs. Mild to moderate myocarditis, hepatitis, splenitis, and interstitial nephritis were also observed in the birds. Immunohistochemistry confirmed Sarcocystis sp. in the capillaries of lungs, hearts, livers, and spleens of most of the birds, but also in the pancreas, kidney, intestine, proventriculus, and brain of a few birds. The probable source of Sarcocystis sp. in these birds was the wild opossum (Didelphis albiventris), a common inhabitant of a local forest that surrounds the Belo Horizonte Zoo (Fundação Zoo-Botânica). This is the first documentation of Sarcocystis infection in psittacines and a pigeon from Brazil.

A study was carried out in order to find out the status of Giardia spp. and Sarcocystis spp. in pet dogs and stray cats of Shiraz, Fars Province of Iran. Faecal samples of 147 pet dogs and 112 stray cats of different age groups, breeds, and sexes were tested. The stools were examined with the following techniques: direct faecal smears using normal saline, zinc sulfate flotation and formalin-ether concentration technique. Out of a total of 147 pet dogs examined, only one case (0.68%) of Giardia spp. was observed. A total of 3 (2.04%) pet dogs were found positive for Sarcocystis spp. Specimens from stray cats were also examined, however no Giardia spp. trophozoite or cyst was observed in these specimens.

Llamas (Lama glama) are South American camelids described as intermediate hosts of Neospora caninum, Toxoplasma gondii and Sarcocystis aucheniae. Due to the potential role of these protozoan infections as a cause of economic losses, the aim of this study was to determine the seroprevalence for T. gondii, N. caninum and Sarcocystis sp. in llamas from Argentina. Serum samples from 308 llamas (>2 years old) were collected between 2005 and 2007. A total of 55 farms located in six departments of Jujuy province, Argentina were sampled. Presence of antibodies to N. caninum, T. gondii and Sarcocystis sp. was determined by the indirect fluorescent antibody test (IFAT). For Sarcocystis, 2 different bradyzoites-based antigens were prepared using S. aucheniae and S. cruzi. Sera were tested at dilutions 1:25 and 1:50. Antibodies to N. caninum were found in 4.6% serum samples. Fifty percent of departments and 14.5% of farms had positive animals. Antibodies to T. gondii were found in 30% of samples, distributed in 66% of departments and 43.6% of farms. Antibodies to Sarcocystis sp. were detected in 96% of samples and all departments and farms had positive animals, suggesting frequent contact between llamas and canids. Co-infection with N. caninum, T. gondii and Sarcocystis sp. was also recorded. Low seroprevalence of N. caninum in llamas detected in this study could be related to climatic and geographical conditions that limit cattle breeding activity, reducing the source of infection for definitive hosts. Seroprevalence of T. gondii and the positive animal distribution suggest frequent contamination of grass with felid faeces. In conclusion, this is the first report of combined seroprevalence for N. caninum, T. gondii and Sarcocystis sp. in llamas. Further studies are needed to determine the potential role of these protozoan infections as cause of abortion in Argentina as well as presence of these protozoans in llama meat used for human consumption.

Samples from the tongue, heart, oesophageal and skeletal muscles were collected from 100 cow and 100 buffalo from Sohag slaughterhouses. Macroscopic examination for sarcocysts was followed by microscopic ones on impression smears and compressed muscles. Histological sections and transmission electron microscope (TEM) studies were done on positive cases. Sarcocystis infection rate of cows were 84%. Two species of Sarcocystis were detected: 1- S. cruzi with a membrane provided with hair-like villar protrusions folded over the surface of the cyst. These protrusions were rod-like, round, oval or irregular in shape and were approximately parallel to the cyst surface. 2-S. hominis characterized by a cyst wall consisting of cylindrical finger-like villar protrusions and having microfilaments. The protrusions were perpendicular on the cyst surface with broad tips and contain microfilaments. Buffaloes' muscle samples revealed an infection rate of 28%. The macroscopic fusiform-shaped species only was detected (Sarcocystis fusiformis). The role of cyst wall ultrastructure and thickness were discussed regarding identification and pathogenicity.

Background: Equine protozoal myeloencephalitis (EPM) is a serious and often fatal neurologic disease of horses, but few studies have investigated risk factors., Objectives: To evaluate operation- and individual-level factors associated with likelihood of the occurrence of EPM., Animals: Data were collected as part of a study of the US equine industry from 1,178 operations representing 83.9% of horses and 51.6% of operations with > or =3 horses in 28 states., Methods: Probability-based sampling was used to enroll representative operations in a cross-sectional study. Interviews were conducted to collect information regarding health and management of horses. A nested case-control study was used to investigate risk factors among individual horses. Interview data were combined with climate data, human population density, and opossum regional ecology categories. Data were analyzed using logistic regression to identify risk factors for the occurrence of EPM., Results: Owners reported that 95% of EPM cases included in this study were diagnosed by veterinarians. Variables associated with EPM occurrence on premises included opossum regional ecology, reported exposure to small wildlife, climate, terrain, housing, choice of bedding material, method of storing feeds, equine stocking density, and primary use of horses. Among individual horses, age was most strongly associated with disease risk. Associations also were identified with sex, breed, primary use, and participation in competitions., Conclusions and Clinical Importance: Because the risk of EPM occurrence on operations is closely tied to factors that impact exposure to opossums, their feces, and their environment, controlling these exposures may be important in preventing the occurrence of EPM.

The range-wide population of American woodcock Scolopax minor has been in slow, steady decline since the late 1960s. The parasite load carried by woodcock and its possible role in the population decline has not been investigated since the early 1970s. A survey of parasites in American woodcock in Connecticut was undertaken in 2002: Sarcocystis spp. was found in 32 of 78 (42%) individuals examined. Elongate sarcocysts, 25 x 125 micro, containing numerous packed bradyzoites with distinct, tightly packed villar projections of the cyst wall, were found scattered throughout skeletal type and myocardial muscle. Sarcocystis spp. was also recorded during the earlier surveys and considered common, but was not examined with the use of electron microscopy. The present study includes the first ultrastructural description of Sarcocystis sp. in the skeletal muscle of woodcock and will serve as a basis for future comparisons in woodcock.

Sarcocystis neurona is an important protozoal pathogen because it causes the serious neurological disease equine protozoal myeloencephalitis (EPM). The capacity of this organism to cause a wide spectrum of neurological signs in horses and the broad geographic distribution of observed cases in the Americas drive the need for sensitive, reliable and rapid typing methods to characterize strains. Various molecular methods have been developed and used to diagnose EPM due to S. neurona, to identify S. neurona isolates and to determine the heterogeneity and evolutionary relatedness within this species and related Sarcocystis spp. These methods included sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immuno-fluorescent assay (IFA), slide agglutination test (SAT), SnSAG-specific ELISA, random amplified polymorphic DNA (RAPD), PCR-based restriction fragment length polymorphism (RFLP), amplified fragment length polymorphism (AFLP) fingerprinting, and sequence analysis of surface protein genes, ribosomal genes, microsatellite alleles and other molecular markers. Here, the utility of these molecular methods is reviewed and evaluated with respect to the need for molecular approaches that utilize well-characterized polymorphic, simple, independent, and stable genetic markers. These tools have the potential to add to knowledge of the genetic population structure of S. neurona and to provide new insights into the pathogenesis of EPM and S. neurona epidemiology. In particular, these methods provide new tools to address the hypothesis that particular genetic variants are associated with adverse clinical outcomes (severe pathotypes). The ultimate goal is to utilize them in future studies to improve treatment and prevention strategies.

Background: Human intestinal sarcocystosis is a zoonotic disease caused by two coccidians, i.e. Sarcocystis fusiformis (syn. S. bovihominis, S. hominis) due to consumption of raw infected beef and Sarcocystis meischeriana (syn. S. suihominis) due to consumption of infected raw pork. In 1987, survey of the macroscopic S. fusiformis cysts in market beef mainly from old water buffalos aged more than 15 years were commonly observed in Bangkok. In 2005, the macroscopic cyst was no longer seen in beef of cattle and water buffalo aged less than three years., Objective: The epidemiological investigation of Sarcocystis spp. infected meat in Bangkok and Lampang., Material and Method: Samples for each of the tongue and beef of cattle and water buffalo, pork from Bangkok markets and pork of domestic swine from some remote villages in various subprovinces (Ampurs) in Lampang were obtained for microscopic examination by H and E and selectively by PAS staining., Results: The microscopic S. fusiformis cysts were seen in all five specimens of tongues and ten specimens of muscles of cattle and water buffalo obtained from fresh-food markets in Bangkok. Ten samples of pork from Bangkok markets revealed no coccidian infection. The microscopic S. meischeriana cysts were seen in three specimens of swine muscles collected from two subprovinces in Lampang., Conclusion: The present merozoites in coccidian cysts retrieved from beef and pork are similar to those previously observed in human intestine. This may histologically indicate an invasive sarcocystosis by both species leading to a condition presently known as chronic inflammation of undetermined etiology in man.

Six Sarcocystis species, i.e. Sarcocystis grueneri, Sarcocystis rangi, Sarcocystis tarandivulpes, Sarcocystis hardangeri, Sarcocystis rangiferi and Sarcocystis tarandi have previously been described from reindeer based on sarcocyst morphology. In order to validate and expand the species descriptions, the complete small subunit (ssu) rRNA gene was sequenced and used to genetically characterise the six species. The aim was to reveal possible genetic variation in the ssu rRNA gene within each Sarcocystis species, between the species from reindeer, but also between the reindeer species and related Sarcocystis species characterised in other studies. Muscle tissue from the heart and diaphragm was sampled from 18 adult semi-domesticated reindeer at a field abattoir in northern Norway. Sarcocysts were excised from the tissue and classified into one of the six known Sarcocystis species based on their morphology. Two cysts of each of the six species from two different animals were randomly selected for further DNA analyses. The complete ssu rRNA gene was amplified by the polymerase chain reaction (PCR) and subsequently sequenced. The complete ssu rRNA gene sequences were used to compare the six species with each other and with other previously sequenced Sarcocystis species retrieved from GenBank. There was little sequence variation between two isolates of the same species, but the six species differed from each other by insertions, deletions and single nucleotide polymorphisms (SNPs), mainly located in variable regions of the gene. The identity between the six species from reindeer was approximately the same as when other Sarcocystis species using a different intermediate host were compared with each other. This study supported previous findings of reindeer being the intermediate host for at least six Sarcocystis species and the results also indicated the existence of certain nucleotide positions within the ssu rRNA gene that are unique to Sarcocystis species with a canine definitive host.

In the current study, we determined the seroprevalence of Toxoplasma gondii, Sarcocystis neurona, and Encephalitozoon cuniculi in three species of lemurs from St. Catherines Island, Georgia. Serum samples were tested from 52 ring-tailed lemurs (Lemur catta), six blue-eyed black lemurs (Eulemur macaco flavifrons), and four black and white ruffed lemurs (Varecia variegata variegata) using an agglutination assay. Three ring-tailed lemurs (5.8%) were positive for T. gondii (titer of 1:50); one ring-tailed lemur (1.9%) and one black and white ruffed lemur (25%) were positive for S. neurona (titers of 1:1000); and one ring-tailed lemur (1.9%) was positive for E. cuniculi (titer of 1:400). All blue-eyed black lemurs were negative for antibodies to T. gondii, S. neurona, and E. cuniculi. This is the first detection of antibodies to T. gondii in ring-tailed lemurs and antibodies to S. neurona and E. cuniculi in any species of prosimian.

A total of 479 stool specimens were collected from rural communities of Ubon Ratchathani Province, Thailand and examined by two techniques: the modified Kato thick smear and the direct smear. The prevalence of Opisthorchis viverrini (14.8%), hookworm (10.2%), Sarcocystis spp (4.6%), Taenia spp (2.9%), Strongyloides stercoralis (2.1%), Giardia lamblia (1.2%), Echinostoma spp (0.6%), Ascaris lumbricoides (0.4%), Entamoeba histolytica (0.2%), Chilomastix mesnili (0.2%) and Endolimax nana (0.2%) were determined. The morphology of the Sarcocystis spp sporocysts examined by both procedures looked similar and was found to be easily recognizable. Among these specimens, 22 cases (4.6%) were positive for Sarcocystis infection detected by the modified Kato technique, whereas only one case (0.2%) was detected by both techniques. These differences were found to be statistically significant (p < 0.05), indicating that the modified Kato technique was decidedly more sensitive than the direct smear procedure in identifying Sarcocystis infection. An epidemiological survey was conducted in Khon Kaen Province involving 1124 stool samples using the modified Kato technique. The greatest frequency was Opisthorchis viverrini at 32.0% while the second highest was Sarcocystis spp at 8.0%. The prevalences of hookworm, Echinostoma spp, Taenia spp, Trichuris trichiura and Enterobius vermicularis were 2.7, 2.1, 1.0, 0.2 and 0.2%, respectively. Other than opisthorchiasis, northeastern Thailand may be an endemic area for sarcocystosis. This is the first report of the applicability and potential usefulness of the Kato thick smear technique for the diagnosis of Sarcocystis infection in a field survey.

This study was undertaken for the determination of Sarcocysts species in sheep slaughtered in Kirikkale Municipality Slaughterhouse. For this study, oesophageal, diaphragm and intercostal muscles were collected from randomly selected 112 sheep out of 1131 sheep (814 sheep and 317 lambs) that were slaughtered from October 2005 to May 2006. The samples were examined for macro and microcyst of Sarcocysts spp. Macrocysts and microcysts were found in 58.92% of the overall samples. Microcysts were found in 47.32% and macrocysts were perceived in 20.53% of the sheep that were under examination. The distribution of the microcysts with respective to the age of the sheep was studied and it has been observed that 16.12% (5 in every 31 lambs) of the lambs under 1 year old and 59.25% (48 in every 81 sheep) of the sheep equal or older than 1 year old had possessed microcysts. Sarcocystis ovicanis (47.32%) and S. arieticanis (1.23%) were the species with the highest and lowest number of recurrences respectively. Macrocysts were observed in every sheep over one age. In addition, randomly selected oesophagi with macrocyst were examined histopathologically. Sarcocystis sporocysts were not found in dog feces.

Neospora caninum, Sarcocystis spp, and Toxoplasma gondii are related coccidian parasites that can cause abortion and neonatal mortality in animals. In addition, T gondii and certain species of Sarcocystis are zoonotic. This article reviews information on the etiology, diagnosis, control, and prevention of these diseases.

The population genetics and systematics of most coccidians remain poorly defined despite their impact on human and veterinary health. Non-recombinant parasite clones characterized by distinct transmission and pathogenesis traits persist in the coccidian Toxoplasma gondii despite opportunities for sexual recombination. In order to determine whether this may be generally true for tissue-cyst forming coccidia, and to address evolutionary and taxonomic problems within the genus Sarcocystis, we characterized polymorphic microsatellite markers in Sarcocystis neurona, the major causative agent of equine protozoal myeloencephalitis (EPM). Bayesian statistical modeling, phylogenetic reconstruction based on genotypic chord distances, and analyses of linkage disequilibrium were employed to examine the population structure within S. neurona and closely related Sarcocystis falcatula isolates from North and South America. North American S. neurona were clearly differentiated from those of South America and also from isolates of S. falcatula. Although S. neurona is characterized by substantial allelic and genotypic diversity typical of interbreeding populations, one genotype occurs with significantly excessive frequency; thus, some degree of asexual propagation of S. neurona clones may naturally occur. Finally, S. neurona isolated from disparate North American localities and diverse hosts (opossums, a Southern sea otter, and horses) comprise a single genetic population. Isolates associated with clinical neurological disease bear no obvious distinction as measured by these presumably neutral genetic markers.

Background: The study reported here was undertaken to assess the presence of antibodies to Sarcocystis neurona in the serum of horses of North American origin that had been relocated for 1 year or more to India (ie, outside of the known endemic areas for S. neurona)., Hypothesis: The presence or absence of such antibodies should provide information concerning the persistence of such antibodies, or support the presence of chronic infection, or both., Animals: A total of 228 Thoroughbred horses were sampled in India, of which 86 were of North American origin that had been in India between 1 and 13 years, 124 were Indian-born horses that had never been out of India, 8 were of Irish origin, 8 were of English origin, and 2 were originally from France., Methods: Sera were tested using established western blot analysis., Results: Of the Indian-born horses, 0.8% were test positive, and of the North American horses, 42% were test positive. All of the English and Irish horses were test negative, and the 2 French horses were test positive., Conclusions and Clinical Importance: These data indicate that antibodies against S. neurona can be detected for many years after horses have been removed from an endemic area and that this may be attributable to long half-life of the antibodies or to chronic infection and ongoing antibody production, or both.

Serum was collected from laboratory-reared Virginia opossums (Didelphis virginiana) to determine whether experimentally infected opossums shedding Sarcocystis neurona sporocysts develop serum antibodies to S. neurona merozoite antigens. Three opossums were fed muscles from nine-banded armadillos (Dasypus novemcinctus), and 5 were fed muscles from striped skunks (Mephitis mephitis). Serum was also collected from 26 automobile-killed opossums to determine whether antibodies to S. neurona were present in these opossums. Serum was analyzed using the S. neurona direct agglutination test (SAT). The SAT was modified for use with a filter paper collection system. Antibodies to S. neurona were not detected in any of the serum samples from opossums, indicating that infection in the opossum is localized in the small intestine. Antibodies to S. neurona were detected in filter-paper-processed serum samples from 2 armadillos naturally infected with S. neurona.

A study was conducted to determine the prevalence and geographical distribution of Sarcocystis spp. infecting 2 subspecies of caribou (Rangifer tarandus) inhabiting Newfoundland and Labrador and its potential definitive hosts. Muscle samples of caribou were obtained, primarily from hunters, and feces of red foxes (Vulpes vulpes) and wolves (Canis lupus), from trappers, and Husky dogs (Canis familiaris), from owners. Histological sections of muscle and flotation methods for feces were used for parasitic detection. Sarcocystis sp. infected more than 50% of barren-ground caribou (R. t. tarandus) from 4 locations in Newfoundland, but it was significantly greater in the north, where 99% of woodland caribou (R. t. caribou) from Labrador harbored the infection. Sporocysts were observed in 27 of 32 red foxes from eastern and northern Newfoundland, whereas 15 of 15 wolves and 22 of the 38 Husky dogs were infected. Wolves and red foxes probably acquired the infection through scavenging, and Husky dogs, from meat they were fed.

Sera from 961 horses from Brazil were tested for antibodies against the major surface antigens SnSAG4 and NhSAG1 to determine the seroprevalence of Sarcocystis neurona and Neospora hughesi, respectively. Antibodies against SnSAG4 were detected in 669 (69.6%) of the horses, while antibodies against NhSAG1 were detected in only 24 (2.5%) of the horses. These serologic results suggest that there is a high concentration of S. neurona in the environment of Brazil, which results in marked exposure of horses to this parasite. Additionally, the data further confirm that infection with Neospora spp. is relatively uncommon in horses.

Monitoring the health of wildlife populations is important for understanding and controlling the risk of infections to livestock, humans and/or other wildlife. In this paper, we analyse the results of surveys of parasites and non-specific signs of diseases carried out on organs from 638 red and 107 sika deer culled in four regions of Scotland between 1991 and 1997. Infections of the lung by Elaphostrongylus spp. were significantly greater in red than sika deer. Older animals were more heavily infected with Elaphostrongylus spp. and Sarcocystis spp., and infections with Sarcocystis spp. tended to be heavier in more recent years. The results suggest that a combination of key indicator parasite species and non-specific signs of disease may be useful for monitoring the health of wildlife populations at a national scale. However, they also demonstrate that such monitoring needs to be long-term, carried out according to standard protocols and at an appropriate resolution to enable integration with data on other potentially influential environmental factors.

The present study examined the seroprevalence of Toxoplasma gondii and Sarcocystis neurona in a population of beavers (Castor canadensis) from Massachusetts. Sixty-two blood samples were collected during the field seasons over 3 consecutive years from different animals. Blood was collected onto filter paper and shipped to the Department of Biomedical Sciences, Virginia Tech, Blacksburg, Virginia, for parasite testing. The samples were tested at dilutions of 1:25, 1:50, and 1:100 against each parasite antigen by modified agglutination tests to determine whether antibodies to either parasite were present in the blood. Six of 62 samples (10%) were positive for T. gondii, with 2 samples having titers of 1:25 and 4 having titers of 1:50. Four of 62 samples (6%) were positive for S. neurona, with 2 samples having titers of 1:25 and 2 having titers of 1:50.

The objectives of the present study were to assess the genetic diversity, phylogeny and phylogeographical relationships of available Sarcocystis neurona isolates from different localities in the United States. All 13 Sarcocystis isolates from different hosts were subjected to polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analyses using two published DNA markers (25/396 and 33/54). The 334 bp sequence of the 25/396 marker of these isolates and Besnoitia darlingi, B. bennetti, Toxoplasma gondii and Neospora caninum were sequenced and compared. Phylogenetic analysis was performed using neighbour-joining (NJ), maximum parsimony (MP) and minimum evolution (ME) methods based on the sequences of the 25/396 marker of the 13 Sarcocystis isolates obtained in this study and sequences of 10 related isolates from GenBank. Phylogenetic trees revealed a close relatedness among S. neurona isolates in the US (nucleotide sequence diversity <5.0%). US isolates formed a monophyletic group and appeared more closely related to each other than to the South American isolates, which formed a separate lineage. NJ and ME trees with Kimura 2-parameter model separated S. neurona into two separate groups: a northern US group and a Southern US group. These findings suggest a correlation between grouping of the isolates and geographical segregation and were consistent with a genetic bottleneck hypothesis during opossum colonisation of North America. These data do not support either the view of S. neurona as a single super-species or its division into multiple subspecies.

The present investigation revealed that the role of natural infection with Sarcocystis in sheep was 84% in Niemy, 76% in Najdy, 89% in Sawakny sheep and 77% in goats slaughtered in Riyadh, Saudi Arabia. The distribution of infection among the examined organs was varied from the sheep strain to goats. Recorded muscle cysts were macroscopic 8-18% (mean 13.5%) and microscopic 73-85% (mean 79%). Macroscopic cysts were usually detected on the oesophagus. EM showed an ultrastructural similarities in the cyst walls and their parasite contents (metrocytes and merozoites) for all macroscopic cysts examined from sheep and goats. These were also enclosed by a secondary cyst wall. The primary cyst wall had many cauliflower-like protrusions supported by many fibellar structures, while the ground substance underneath them contained many dark, dense granules and usually extended into the cyst cavity dividing it into many compartments containing the parasites. The cyst parasites were the peripheral globular to spherical metrocytes, which were divided by endodyogeny and the central banana-shaped merozoites. Both metrocytes and merozoites had all characteristics of the Apicomplexa. Experimental transmission of Sarcocystis to the definitive host, revealed that kittens were the only suitable final host. Thus, gamogony and zygote formation (fertilization) occurred 2-8 days post infection (p.i.) while sporulation took place within this final host 13-15 days p.i., where sporocysts were passed within faeces of infected kittens at that time. Also, the study indicated that the prepatent period was 13-15 days, while the patent period was 37-42 days.

We investigated the prevalence of sarcocystosis in 826 goats slaughtered in the winter season from November to April in northern Iraq. The prevalence of macrocysts was on average 34%, with only 20% infected animals in November, but 46% in February. The infection rate in 1-, 3- and 6-year-old goats was 4%, 48%, and 83%, respectively. The highest specificity of infection was in the oesophagus (99%) and the lowest in the diaphragm (3%). Grossly, we identified 2 forms of macroscopic sarcocysts, fat and thin, with different morphological characteristics. The prevalence of microcysts was 97% and no effects of age, sex and seasonal variations were observed. Development of microcysts in the small intestine of dogs and cats has also been investigated. The pre-patent period in experimentally infected dogs was 12-14 days and the patent period lasted 64-66 days. A dog shed about 155 million sporocysts, but no sporocysts were shed by cats that had been fed the same infected tissues, thus identifying the microcysts as Sarcocystis capracanis.

A cross-sectional survey was performed to estimate the prevalences of antibodies to Toxoplasma gondii (ELISA, IFAT), Sarcocystis spp. (ELISA, using S. miescheriana as antigen) and Neospora caninum (ELISA, immunoblotting) in sows from breeding farms in southern Hesse, Germany. A total of 2041 plasma samples of sows from 94 randomly selected farms was examined. Data on farm profiles, husbandry management and sows were collected by a questionnaire and exploratively analysed. For T. gondii the ELISA results agreed well with the results obtained by IFAT (kappa=0.71). Antibodies to T. gondii were detected by ELISA in 19% of the sows. Sixty-nine percent of the farms had at least one seropositive sow, and a within-farm seroprevalence of >or=50% was observed in 14% of all farms. The prevalence of anti-T. gondii antibodies was positively correlated with the age of sows. The within-herd seroprevalence was significantly higher in farms with reproductive disorders than in those without such problems. On the farm level, the farm type 'piglet production' (versus 'pedigree breeding' or 'farrow-to-finish') was the only risk factor associated with the presence of T. gondii-seropositive sows. Antibodies to Sarcocystis spp. were found in 29% of the sows. Seventy-two percent of the farms harboured at least one seropositive sow, and a within-farm seroprevalence of >or=50% was detected in 23% of all farms. The seroprevalence increased significantly with the age of sows. On the farm level, only the farm type 'piglet production' (versus 'pedigree breeding') and the replacement of sows by purchasing (versus raising on the own farm) were identified as risk factors for seropositivity. Antibodies to N. caninum were detected in one sow using both the screening ELISA and the confirmatory immunoblotting technique. This may indicate the first natural N. caninum infection in pigs.

The study objective was to assess the risk of transplacental transmission of Sarcocystis neurona and Neospora hughesi in foals from 4 California farms during 3 foaling seasons. Serum of presuckle foals and serum and colostrum of periparturient mares were tested using indirect fluorescent antibody tests for S. neurona and N. hughesi. Serum antibody titers were < or =10 in 366 presuckle foals tested. There was no serologic or histologic evidence of either parasite in aborted fetuses or placentas examined. Positivity for S. neurona and N. hughesi in mares increased with age. Mares < or =9 yr that originated from Kentucky were 3.8 and 1.4 times more likely to be positive for S. neurona and N. hughesi, respectively, than mares from California. The strength of association between positivity to either parasite and state of birth decreased as age increased. Mares positive for S. neurona and N. hughesi were 2.2 and 1.7 times more likely, respectively, to have a previous abortion than negative mares, adjusted for age and state of birth. The annual mortality rate for mares was 4%. The annual incidence rate of equine protozoal myeloencephalitis was 0.2%. In conclusion, there was no detectable risk of transplacental transmission of S. neurona and N. hughesi. Prevalence of antibodies against both parasites in mares increased with age.

From April 1996 to December 2002 the prevalence of Sarcocystis neurona sporocysts in North American opossum (Didelphis virginiana) in Southern Michigan was estimated. Sporocysts of S. neurona were found in intestinal scrapings from 31 (15%) of 206 examined opossum. The frequency of infection was higher in adult animals (26/206; 12.6%) and females (19/206; 9.2%) than in juveniles (5/206; 2.4%) and males (12/206; 5.8%). Also, prevalence of S. neurona sporocysts in opossums in relation to factors such as age, sex, season, body condition, presence of concomitant infection, and presence of young in the pouch of females was studied in detail over the course of the year, 2002. Univariate analyses identified the following factors as being associated with the presence of S. neurona sporocysts in opossums: (i) for age, adult (odd ratio [OR] = 2.074, P = 0.0005); (ii) for sex, female (OR = 7.016, P = 0.0119); (iii) for season, summer (OR = 7.917, P = 0.0032) and spring (OR = 4.071, P = 0.1063); (iv) for body condition, poor (OR = 3.50, P = 0.1200) and good (OR = 1.167, P = 0.8637); (v) for the presence of concomitant infection (OR = 23.056, P = 0001), and (vi) for the presence of young in the pouch of females (OR = 40.083, P = 0.0001). Multivariate logistic-regression analyses selected the following factors as being significantly associated with presence of S. neurona sporocysts in opossums: (i) for the presence of concomitant infection (OR = 8.722, P = 0.0160) and (ii) for the presence of young in the pouch of females (OR = 31.915, P = 0.0065). The prevalence of S. neurona sporocysts in D. virginiana suggests that this opossum may constitute an ample reservoir of infection to other animals in the northern United States.

Sarcocystis species are intracellular protozoan parasites with an intermediate-definitive host life cycle based on a prey-predator relationship. Asexual stages develop in intermediate hosts after they ingest the oocyst stage from definitive-host feces and terminate with the formation of intramuscular cysts (sarcocysts). Sarcocysts in meat eaten by a definitive host initiate sexual stages in the intestine that terminate in oocysts excreted in the feces. Most Sarcocystis species infect specific hosts or closely related host species. For example, humans and some primates are definitive hosts for Sarcocystis hominis and S. suihominis after eating raw meat from cattle and pigs, respectively. The prevalence of intestinal sarcocystosis in humans is low and is only rarely associated with illness, except in volunteers who ingest large numbers of sarcocysts. Cases of infection of humans as intermediate hosts, with intramuscular cysts, number less than 100 and are of unknown origin. The asexual stages, including sarcocysts, can stimulate a strong inflammatory response. Livestock have suffered acute debilitating infections, resulting in abortion and death or chronic infections with failure to grow or thrive. This review provides a summary of Sarcocystis biology, including its morphology, life cycle, host specificity, prevalence, diagnosis, treatment, and prevention strategies, for human and food animal infections.

Equine protozoal myeloencephalitis is the most important protozoan disease of horses in North America and is usually caused by Sarcocystis neurona. Natural and experimentally induced cases of encephalitis caused by S. neurona have been reported in raccoons (Procyon lotor) and raccoons are an intermediate host for this parasite. A 3-yr-long serological survey was conducted to determine the prevalence of agglutinating antibodies to S. neurona in raccoons collected from Fairfax County, Virginia, a suburban-urban area outside Washington, D.C. Samples from 469 raccoons were examined, and agglutinating antibodies (> or = 1:50 dilution) were found in 433 (92.3%) of the raccoons. This study indicates that exposure to S. neurona is high in this metropolitan area.

A total of 206 Virginia opossums ( Didelphis virginiana) collected from the mid-Michigan region, United States, during a period extending from 1996 to 2002 were sampled for the presence of Sarcocystis spp sporocysts. All isolates were phenotypically identified as Sarcocystis spp and genotyped to the species level by PCR-based techniques. The overall prevalence of Sarcocystis spp in opossums was 18% (37/206). The prevalence of Sarcocystis spp differed significantly with age ( P<0.001) and adult opossums were more commonly infected (14.6%; 30/206) than juveniles (3.4%; 7/206). No significant difference in the prevalence of Sarcocystis spp infection was observed between male and female ( P<0.15). The highest prevalence was recorded during summer (9.2%; 19/206). PCR-RFLP analyses demonstrated the majority of Sarcocystis isolates to be S. neurona, with some animals co-infected with sporocysts of S. falcatula. Out of the 37 Sarcocystis-infected opossums, 23 (62%) had sporocysts of S. neurona only, four (11%) had sporocysts of S. falcatula only, and eight (22%) had a mixture of S. neurona and S. falcatula sporocysts. These findings indicate that mixed Sarcocystis infections in opossums are common. The propensity for Sarcocystis spp to co-exist in the opossum gut enhances dissemination and environmental contamination with these coccidia. Additionally, this increases the chance for sexual recombination between Sarcocystis spp, given the proclivity of these species to reproduce sexually at high numbers in the intestinal cells of their definitive host.

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies to Sarcosystis infection in cattle and buffaloes. Crude antigens derived from cystozoites of Sarcosystis cruzi and antibodies from the sera of naturally infected cattle and buffaloes were used. The assay was validated by determination of the specificity, sensitivity and negative and positive predictive values. Using ELISA, a sero-epidemiological survey for sarcosystosis was carried out in cattle (n = 215) and buffaloes (n = 123) from the wet and dry zones of Sri Lanka. The prevalence of infection in cattle was 69.3%. Prevalence of the infection collectively in both cattle and buffaloes in the dry zone was significantly higher (P < 0.001) than that in the wet zone. The number of sero-positive cattle and buffaloes in the dry zone was significantly higher than that in the wet zone (P < 0.05). Buffaloes in the dry and wet zones showed a greater prevalence of infection (P < 0.05) than cattle.

Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease in the horse most commonly caused by Sarcocystis neurona. The domestic cat (Felis domesticus) is an intermediate host for S. neurona. In the present study, nine farms, known to have prior clinically diagnosed cases of EPM and a resident cat population were identified and sampled accordingly. In addition to the farm cats sampled, samples were also collected from a mobile spay and neuter clinic. Overall, serum samples were collected in 2001 from 310 cats, with samples including barn, feral and inside/outside cats. Of these 310 samples, 35 were from nine horse farms. Horse serum samples were also collected and traps were set for opossums at each of the farms. The S. neurona direct agglutination test (SAT) was used for both the horse and cat serum samples (1:25 dilution). Fourteen of 35 (40%) cats sampled from horse farms had circulating S. neurona agglutinating antibodies. Twenty-seven of the 275 (10%) cats from the spay/neuter clinic also had detectable S. neurona antibodies. Overall, 115 of 123 (93%) horses tested positive for anti-S. neurona antibodies, with each farm having greater than a 75% exposure rate among sampled horses. Twenty-one opossums were trapped on seven of the nine farms. Eleven opossums had Sarcocystis sp. sporocysts, six of them were identified as S. neurona sporocysts based on bioassays in gamma-interferon gene knockout mice with each opossum representing a different farm. Demonstration of S. neurona agglutinating antibodies in domestic and feral cats corroborates previous research demonstrating feral cats to be naturally infected, and also suggests that cats can be frequently infected with S. neurona and serve as one of several natural intermediate hosts for S. neurona.

A sampling of equids from the state of Oklahoma produced an estimate of seroprevalence of antibody to Sarcocystis neurona to be about 89.2%. This figure represents the highest currently reported regional seroprevalence of antibody to this organism. Regional differences in seroprevalence were found in the western quadrants of the state relative to the eastern quadrants of the state, with a significantly higher seroprevalence in the eastern regions. Thoroughbreds were found to exhibit a statistically significant lower seroprevalence as a breed group when compared with other breeds sampled.

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).

Sarcocystis neurona, Neospora caninum, N. hughesi, and Toxoplasma gondii are 4 related coccidians considered to be associated with encephalomyelitis in horses. The source of infection for N. hughesi is unknown, whereas opossums, dogs, and cats are the definitive hosts for S. neurona, N. caninum, and T. gondii, respectively. Seroprevalence of these coccidians in 276 wild horses from central Wyoming outside the known range of the opossum (Didelphis virginiana) was determined. Antibodies to T. gondii were found only in 1 of 276 horses tested with the modified agglutination test using 1:25, 1:50, and 1:500 dilutions. Antibodies to N. caninum were found in 86 (31.1%) of the 276 horses tested with the Neospora agglutination test--the titers were 1:25 in 38 horses, 1:50 in 15, 1:100 in 9, 1:200 in 8, 1:400 in 4, 1:800 in 2, 1:1,600 in 2, 1:3,200 in 2, and 1:12,800 in 1. Antibodies to S. neurona were assessed with the serum immunoblot; of 276 horses tested, 18 had antibodies considered specific for S. neurona. Antibodies to S. neurona also were assessed with the S. neurona direct agglutination test (SAT). Thirty-nine of 265 horses tested had SAT antibodies--in titers of 1:50 in 26 horses and 1:100 in 13. The presence of S. neurona antibodies in horses in central Wyoming suggests that either there is cross-reactivity between S. neurona and some other infection or a definitive host other than opossum is the source of infection. In a retrospective study, S. neurona antibodies were not found by immunoblot in the sera of 243 horses from western Canada outside the range of D. virginiana.

Investigations to occurrence of Sarcocystiosis of domestic animals in Algeria were carried out by different methods--Immunofluorescence (IFAT), macroscopic and microscopic examinations of muscle, coproscopia. Positive results could be found in intermediate hosts--362 of 537 cattle, 188 of 292 sheep, 13 of 57 horses and in the final hosts dogs and cats. Muscle-probes from cattle contained S. cruzi (58.7 respectively 61.0%), S. hirsuta (48.9 respectively 35.0%), from sheep S. tenella (61.0 respectively 59.6%, S. arieticanis (38.8 respectively 40.3%), from horses S. bertrami (24.5%). IFAT-data of extensy: 64.5% in cattle, 60.5% in sheep. First time in Algeria could be found S. hominis (2.3 respectively 3.9%).

Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by infection of the central nervous system with the protozoan parasite Sarcocystis neurona. A herd-level analysis of a cross-sectional study of serum antibodies to S. neurona in Michigan equids was conducted, using data collected in 1997 for study that included 1121 equids from 98 Michigan horse farms. Our objective was to identify specific herd-level risk factors associated with seropositivity. We tested associations between herd seroprevalence and various farm-management practices (including feed-storage methods and wildlife control). Multivariable models were developed for three strata based on relative opossum abundance (opossum districts). Herd seroprevalence ranged from 0 to 100% (median=57%). No risk factor was significantly associated with herd seroprevalence at P< or = 0.05 in all opossum districts. Our results suggest that equids living in areas with large opossum populations might be infected with S. neurona from multiple sources.

Sarcocystis infection was detected in 93% of horses in Mongolia. Using the compress method, sarcocysts were found in the muscles of the diaphragm, heart and tongue in 40 of the 43 horses that were slaughtered at the Makh Impex Meat Company in Ulaan Baatar in July 1998. The muscle of the tongue showed the highest rate (97.5%) of infection. The distribution of sarcocysts in the muscles was positively correlated with horse age; the rate of detection was significantly lower (p=0.01) in the under 10 year old group than the older group. All horses were apparently healthy and were slaughtered for human consumption.

Parasite Biology, Epidemiology and Systematics Laboratory, Animal and Natural Resources Institute, Agricultural Research Service, United States Department of Agriculture, Building 1001, Beltsville, Maryland 20705-2350 Antibodies to Neospora caninum and Sarcocystis neurona were determined in serum samples of 502 domestic cats from Brazil using direct agglutination tests with the respective antigens. Antibodies to S. neurona were not found in 1:50 dilution of any serum in the S. neurona agglutination test. suggesting that domestic cats from São Paulo city were not exposed to S. neurona sporocysts from opossums. Antibodies to N. caninum were found in 60 (11.9%) of 502 cats with titers of 1:40 in 36 cats, 1:80 in 18 cats, 1:160 in 5 cats, and 1:800 in 1 cat using the Neospora agglutination test (NAT). Antibodies to N. caninum were confirmed by Western blotting in the sera of 10 cats with NAT titers of 1:80 to 1:800; this finding suggests that at least 10 cats had N. caninum-specific antibodies confirmed by 2 tests. This is the first documentation of natural exposure of cats to N. caninum.