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Muscles of 25 bobcats (Lynx rufus) from remote areas of Mississippi in 2017 were tested for parasites. Testing for Sarcocystis infections included microscopic examination of fresh unstained muscle squashes, pepsin digestion of hearts and tongues, and histological sections of paraffin-embedded tissues. Sarcocystis spp. infections were detected in the muscles of 21 (84%) by a combination of methods. Sarcocysts were detected in the unstained tongue squashes of 2 bobcats. Sarcocystis sp. bradyzoites were detected in the pepsin digests of 3 of 19 hearts, and 12 of 19 tongues. In paraffin-embedded histological sections, sarcocysts were detected in 7 of 25 hearts, 17 of 25 tongues, and 5 of 23 limb muscles. Based on the character of the cyst wall, at least 3 morphologic types of sarcocysts were detected: those with small spikes on the cyst wall, corresponding to Sarcocystis felis, those with long villar protrusions, corresponding to Sarcocystis neurona, and those lacking visible cyst wall protrusions, representing an unidentified type of sarcocyst. Myositis associated with sarcocysts was seen in the tongues of 3, and in the limb muscles of 1 bobcat. Multilocus genotyping of the DNA extracted from paraffin-embedded sections from 2 bobcats, employing 18S, 28S, COI, ITS-1, and 5.8S and rpoB genes, diagnosed Sarcocystis caninum, S. felis, Sarcocystis lutrae, and S. neurona. An encapsulated species of Trichinella was identified in the tongue of 1; it represents the first documented occurrences in bobcats from Mississippi. Taken together, these observations suggest intensive exposure of these wild carnivores to Trichinella tissue cysts, implies predation or scavenging on these tissues promotes parasite transmission, and raises caution concerning zoonotic risk when such meat is rendered for human consumption., (© American Society of Parasitologists 2024.)

This study aimed to estimate the prevalence and distribution patterns of Sarcocystis spp. in cattle tissues in Chachapoyas province in the Peruvian tropical Andes. Additionally, the risk factors associated with the prevalence and the correlation of two diagnostic techniques (direct microscopy of squashed fresh muscle tissues and histopathology) were explored. The tongue, heart, esophagus, Latissimus dorsi muscle, and diaphragm of 210 animals slaughtered in the municipal slaughterhouse of Chachapoyas were evaluated by both techniques. Macroscopic sarcocysts were detected in 16.7% of tissues (CI 95% 11.7-21.7%). The total prevalence of Sarcocystis spp. was 96.2% (95% CI 93.6-98.8%) by direct light microscopy and 100% by histopathology. The highest Sarcocystis prevalence was detected in the esophagus. No significant statistical differences were found in the prevalence of Sarcocystis related to sex, age, or provenance. Both techniques demonstrated a very weak Kappa correlation (κ ≤ 0.24) in predicting the presence of the parasite in each of the five evaluated muscles. Direct microscopy can be implemented at slaughterhouses as a rapid screening test, but it is essential to confirm by histopathology the absence of the parasite in direct-microscopy-negative samples. It is also recommended that beef from the Peruvian Andes be thoroughly cooked for both human and animal consumption because of the zoonotic potential of some species of Sarcocystis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Cattle and buffaloes, popular protein sources worldwide, are intermediate hosts for several Sarcocystis species. These coccidian protozoans cause sarcocystosis resulting in subclinical and chronic infections in striated muscles by forming macrocysts or microcysts. In Thailand, Lao People's Democratic Republic, and Cambodia, Sarcocystis species have been reported, but molecular identification has been lacking. This study investigates the prevalence of infection, histo-morphology, and molecular identification of Sarcocystis species in hearts of cattle and buffalo sold in local markets. A phylogenetic tree inferred from a portion of the 18S ribosomal (r) RNA gene was used to identify the genus and species of Sarcocystis. The mitochondrial cytochrome c oxidase subunit 1 (cox-1 gene) was sequenced to confirm the species of host tissue. In Thailand, Sarcocystis was detected in 66.7% (14/21) of samples. In Lao People's Democratic Republic, 90% (9/10) of samples were infected and in Cambodia 100% (8/8). For the first time from these countries, we report Sarcocystis cruzi, Sarcocystis heydorni, and Sarcocystis levinei found in taurine cattle (Bos taurus) and water buffalo (Bubalus bubalis). Zoonotic protozoan transmission needs to be controlled by inspection activities by local health inspectors, and appropriate action is required at all points in the food chain by competent authorities to protect consumer health and prevent sarcocystosis in cattle and water buffaloes., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Purpose: Sarcocystis spp. are protozoan parasites of livestock which also infect birds, lower vertebrates and mammals, including man. Wild and domestic ruminants such as red deer, roe deer, fallow deer, cattle, sheep and goats may act as intermediate hosts for many Sarcocystis species, some of which are significant pathogens causing sarcocystosis in livestock and humans. The purpose of the present study was to determine the prevalence of Sarcocystis species in fallow deer farmed in an open pasture system., Methods: Samples of heart and oesophagus tissue taken from five fallow deer were examined by light microscope for the presence of sarcocysts. Genomic DNA was extracted from individual sarcocysts. ssu rRNA was successfully amplified using their DNA as templates., Results: Analysis of the ssu rRNA identified the presence of two S. morae sarcocysts in the heart tissue; similarly, S. gracilis sarcocysts were identified in the heart and oesophagus, and Sarcocystis sp. most closely related to S. linearis and S. taeniata were detected in oseophagus., Conclusions: These findings confirm the presence of Sarcocystis spp. in farmed fallow deer in Poland; however, more molecular studies are needed.

Objective: Sarcocystosis is an important zoonotic protozoal disease with worldwide distribution and wide range of hosts. The aim of the present study was to determine the intensity of Sarcocystis spp. infection and to show histopathological features of their cystic lesions in slaughtered cattle of Zabol- Iran., Methods: From April to September 2018, 500 tissue samples from esophagus, heart, diaphragm, tongue and masticatory muscles were prepared from 100 slaughtered cattle. All samples were fixed in 10% neutral buffered formalin and routine tissue processing protocol was performed., Results: The microscopic results showed that 92.2% of specimens had thin-walled cysts of S. cruzi and 14% had thick-walled Sarcocystis ( S. hirsuta and S. hominis ) but macrocyst was only observed in one cattle. The positivity rate of thin walled cysts was 58.8% for heart, 13.9% for masticatory muscles, 10.2% for tongue, 9.3% for esophagus and 7.8% for diaphragm. The positivity rate of thick walled cysts was 32.8% for esophagus, 28.6% for tongue, 22.9% for heart, 15.7% for masticatory muscles and 0% for diaphragm, which could represent either S. hominis or S. hirsuta . The most infected tissue was heart and the least infected tissue was diaphragm. Thin walled cysts ( S. cruzi ) were mostly found in heart and were less found in diaphragm. However, thick-walled cysts ( S. hirsuta and S. hominis ) were mostly detected in esophagus. No thick-walled cysts were found in diaphragm muscle., Conclusion: A high positivity rate of sarcocystosis in slaughtered cattle in Zabol abattoir revealed heavily environmental contamination of Sistan region by this important parasitic disease.

The aim of this study was to investigate the occurrence of sarcocysts in sheep slaughtered in the Rio Grande do Sul state, Brazil. Heart and esophagus samples from 130 sheep were subjected to macroscopic and microscopic examination, followed by molecular analysis. Ten sheep (7.7%) had Sarcocystis gigantea macrocysts in esophagus, as identified by gene sequencing. Microcysts were present in 96.1% of the sheep, with a higher frequency (p < .05) in the heart (91.5%) compared to the esophagus (81.5%) samples. The microcysts were identified as Sarcocystis tenella by gene sequencing. Our results revealed a high frequency of Sarcocystis spp. infection in sheep from southern Brazil. To the authors knowledge, this is the first molecular confirmation of S. gigantea presence in Brazil., (Copyright © 2018. Published by Elsevier B.V.)

Myocardium and diaphragm samples of cattle (n = 521) from HeNan Province (China) were screened for Sarcocystis sarcocysts by histological examination, pepsin digestion, and molecular assays. Morphology and molecular assays were used for identification. The prevalence of Sarcocystis infection in cattle was 41.5% (216/521). Histological examination identified sarcocysts in the myocardium (49.4%, 200/405) and diaphragm (13.8%, 16/116) of cattle. Two species were identified, namely S. cruzi (41.3%, 215/521) and S. hominis (0.2%, 1/521). The findings of the present study indicate a high prevalence of S. cruzi infection in cattle from central China., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Sarcocystis spp. are ubiquitous protozoan parasites that can form cysts in striated muscle and CNS of cattle. Cattle hearts are commonly infected by microscopic sarcocysts. Humans can get infected by consuming cattle meat containing the zoonotic parasites Sarcocystis hominis and Sarcocystis heydorni. However, bovine myocardium is generally infected by Sarcocystis cruzi. The aims of this study were to investigate the occurrence of sarcocysts and the identity of Sarcocystis species present in cattle hearts destined to human consumption in the Central region of the Rio Grande do Sul state, southern Brazil. A total of 314 cattle myocardium samples collected from a local abattoir were microscopically examined for the presence of sarcocysts. The sarcocysts isolated from 134 of these samples (ten sarcocysts per sample) were subjected to DNA extraction and PCR amplification. The PCR-amplified DNA fragments were digested with the restriction enzymes BclI and RsaI (PCR-RFLP) for differentiation among S. cruzi, S. hirsuta, and S. hominis. Sarcocystis species identification was confirmed using DNA sequencing of the cox1 mitochondrial DNA. Sarcocysts were detected in all the bovine myocardium samples. PCR-RFLP analysis resulted in successful amplification of 78 of the 134 samples tested. Only the S. cruzi DNA restriction pattern was identified from all of the 78 amplified samples. DNA sequencing also confirmed the presence of S. cruzi DNA. In conclusion, all myocardium samples evaluated were infected with microscopic sarcocysts. S. cruzi was the only species detected infecting the cattle hearts., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The cysts of Sarcocystis grueneri were detected and characterized from the cardiac muscles of the Korean water deer ( Hydropotes inermis argyropus ). Of the 38 heart muscle samples examined by light microscopy, 10 were found infected with the cysts of Sarcocystis sp. The cysts appeared oval to spherical shape and measured 110-380 μm in length and 90-170 μm in width. A phylogenetic tree of the 18S rRNA sequences (1.5 kb) revealed a close relationship of the infected cysts to genus Sarcocystis . The 18S rRNA sequence of the infected cysts showed 100% identity to S. grueneri and 97% to S. capracanis . Here, we first report the S. grueneri infections in the Korean water deer.

To survey the prevalence of Sarcocystis infections, 210 heart samples were collected from Korean native cattle ( Bos taurus coreanae ) at an abattoir in Daejeon Metropolitan City, Republic of Korea. Sarcocysts were detected form 31 specimens (14.8%) and identified as Sarcocystis cruzi via transmission electron microscopy. The wall of S. cruzi has flattened protrusions that did not contain fibrils or microfilaments. The protrusions arose irregularly from the base, contained a fine granular substance, lacked internal microfilaments, and measured approximately 0.21-1.25 μm in length and 0.05-0.07 μm in width. Sequence analysis revealed 99.5% homology to S. cruzi . This is the first report on the prevalence of S. cruzi in native cattle from the Republic of Korea.

A 3-year-old sheep was examined after an acute onset of hind limb paralysis and ataxia. At necropsy, central nervous system, pulmonary and intestinal hyperaemia and ecchymoses in the aortic arch were observed. Main microscopic lesions were confined to the heart, cerebrum and cerebellum. There were a multifocal mild myocarditis and nonsuppurative meningoencephalitis together with protozoal cysts in the heart and the brain. Protozoal cystic structures were observed within many of the myocardial fibers as well as in the cerebrum and cerebellum. Using light microscopy it could not be morphologically determined whether these organisms were Toxoplasma (T.) gondii or Neospora (N.) caninum. Additional diagnostic methods like immunohistochemistry and polymerase chain reaction provided differentiation of Sarcocystis from T. gondii and N. caninum. Transmission electron microscopy demonstrated characteristic features of Sarcocystis sp. as previously described. This is the first confirmed diagnosis of Sarcocystis sp. in the central nervous system of a sheep from Turkey.

Molecular detection of Sarcocystis spp. in tissue samples can be useful for experimental and diagnostic purposes. However, the parasite spreads unevenly through tissues, forming tissue cysts, and the cystic wall is an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the cysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of four protocols for DNA extraction from cysts of Sarcocystis spp. present in bovine myocardium samples or after their harvest in phosphate-buffered saline (PBS) solution as well as determine the effects of single or multiple sampling on the accuracy of molecular diagnosis of sarcocystosis in cattle hearts. Cysts and myocardium samples from nine bovine hearts were randomly distributed to four DNA extraction protocols: kit, kit with modification, DNAzol, and cetyl-trimethyl ammonium bromide (CTAB). Samples were submitted to DNA extraction and PCR as replicates of each heart (simplicate, duplicate, and triplicate), and the probability of a true positive diagnostic was calculated. Among the protocols tested, the kit with modification was determined to be the most suitable for DNA extraction from cysts in PBS solution (92.6 % of DNA detection by PCR); DNAzol resulted in higher DNA detection frequency from bovine myocardium samples (48.1 %). Multiple sampling improved the molecular diagnosis of Sarcocystis spp. infection in cattle hearts, increasing at 22.2 % the rate of true positive diagnostic.

Sarcocystis spp. are protozoan parasites with a heteroxenous life cycle, which produce cysts in the muscle of herbivorous animals. In these animal species, sarcocystosis is frequently asymptomatic, although it may occur with high prevalence. Seven Sarcocystis spp. have been described in red deer (Cervus elephus). The aim of this study was to determine the prevalence of sarcocystosis, and to perform the morphological and molecular characterization of Sarcocystis spp. found in wild red deer of the Nahuel Huapi National Park (NHNP), Patagonia, Argentina. Full necropsies of 62 red deer killed by hunters in the NHNP and neighboring areas were performed. Samples of heart and skeletal muscle were examined histologically and selected samples were also examined by transmission electron microscopy (TEM), PCR and sequencing. Sarcocystis spp. thin walled cysts were detected in 62 % (38/62) of heart, and in 22 % (3/14) of skeletal muscle samples examined histologically. TEM revealed a smooth and thin cyst wall (≤1 μm), with scarce and separated ribbon-like protrusions. A total of three partial and one full 18S ribosomal RNA (rRNA) gene sequences were obtained, and showed the highest identity (≥99 %) with Sarcocystis taeniata, a species described in moose (Alces alces). The morphological and molecular results indicate that red deer in Argentina are frequently infected with S. taeniata, a species for which the definitive host is unknown. The present results also confirm that Sarcocystis spp. using cervids as intermediate host are not host-specific. Further studies are needed to improve the epidemiological knowledge of Sarcocystosis in red deer.

Three species of Sarcocystis-S. miescheriana, S. suihominis, and S. porcifelis-have been recorded from pigs ( Sus scrofa ). Among these 3 species, the zoonotic species S. suihominis is of paramount importance and an important food safety issue. Previous studies indicate prevalence of porcine Sarcocystis species in India, but molecular evidence, among other evidence, is required for proper species differentiation. Myocardium from 250 stray and farm pigs destined for slaughter for human consumption were collected from slaughter shops located in urban slums in Punjab, northern India. Tissues were examined for Sarcocystis by using an intact cyst isolation method, pepsin acid digestion, Sarcocystis 18S ribosomal RNA polymerase chain reaction (PCR), and real-time quantitative PCR with melting curve analysis (qPCR-MCA). The combination of primers was used for 18S rRNA PCR amplification followed by sequencing. Ten representative samples were sequenced in both the directions from which 7 readable sequences were obtained for phylogenetic analysis. Sarcocystis cysts/zoites were recorded in 146 (58.4%), 169 (67.6%), 182 (72.8%), and 191 (76.4%) of samples by using intact cyst isolation, pepsin HCl digestion, conventional PCR, and qPCR-MCA, respectively. Molecularly, 1 S. miescheriana isolate and 6 isolates of the zoonotic species S. suihominis were recorded. This is the first study providing molecular identification for the presence of zoonotic species S. suihomonis in India. The prevalence of zoonotic S. suihominis in pork in India is worrisome and warrants intervention policies to stop the practice of rearing pigs under unhygienic conditions.

In the present study, the heteroxenous life cycle of Sarcocystis species from three strains of the slaughtered sheep at Al-Azizia and Al-Saada abattoirs in Riyadh city, K.S.A., was studied. Muscle samples of the oesophagus, diaphragm, tongue, skeletal and heart muscles were examined. Varied natural infection rates in the muscles of the examined sheep strains were recorded as 83% in Niemy, 81.5% in Najdy and 90% in Sawakny sheep. Muscles of the diaphragm showed the highest infection level above all organs except Najdy sheep in which oesophagus has the highest rate. Also, the heart was the lowest infected organ (40% Niemy, 44% Najdy and 53% Sawakny). Microscopic sarcocysts of Sarcocystis arieticanis are easily identified in sections through the heart muscles of the domestic sheep Ovis aries (Artiodactyla: Bovidae). Cysts measured 38.5-64.4 μm (averaged 42.66 μm) in width and 62.4-173.6 μm (averaged 82.14 μm) in length. The validity of this species was confirmed by means of ultrastructural characteristics of the primary cyst wall (0.1-0.27 μm thick) which revealed the presence of irregularly shaped crowded and hairy-like projections underlined by a thin layer of ground substance. This layer consisted mainly of fine, dense homogenous granules enclosing the developing metrocytes and merozoites that usually contain nearly all the structures of the apical complex and fill the interior cavity of the cyst. Several septa derived from the ground substance divided the cyst into compartments. The merozoites were banana-shaped and measured 12-16 μm in length with centrally or posteriorly located nuclei. Experimental infection of carnivores by feeding heavily infected sheep muscles revealed that the dog, Canis familiaris, is the only final host of the present Sarcocystis species. Gamogony, sporogonic stages and characteristics of sporulated oocysts were also investigated.

Muscular samples from the oesophagus, diaphragm and heart of 101 roe deer (Capreolus capreolus) hunted in Galicia (Northwestern Spain) were examined, by the compression method, for the presence of Sarcocystis spp. infection. The structure of the cyst wall was examined by light (LM) and transmission electron microscopy (TEM). The overall prevalence of infection was very high (99%), with a density of 404 cysts/sample (SD 812). The prevalence was very similar in the different examined muscle types (99% heart and diaphragm, and 98.9% oesophagus). A significantly higher intensity of infection was found in the heart (831; SD 1281), followed by the diaphragm (197; SD 190) and the oesophagus (180; SD 205). Macrocysts (>1500 μm long) were only detected in the oesophagus of 48.5% of the examined roe deer; their mean size was 2055.4 μm (SD 632.46). Cysts localised in the myocardium were significantly shorter (371.5 μm; SD 160.47) than those found in the diaphragm (678.2; SD 546) and the oesophagus (973.4 μm; SD 667.87). By LM, most of the cysts (98.8%) displayed a thin wall, which was consistent with those of Sarcocystis sp., S. gracilis and S. capreolicanis; only 1.2% of the cysts had a thick striated wall, consistent with Sarcocystis silva. Three morphological distinct sarcocysts were observed by TEM: the unnamed species Sarcocystis sp., S. capreolicanis and S. gracilis. The wall ultrastructure of the examined macrocysts was consistent with S. gracilis. This study has revealed that Spanish roe deer harbours 4 morphologically distinct types of sarcocysts; being the first record of S. gracilis in roe deer from Spain., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Experiments have established that the first target for echinococcus is the liver and lung and that for pathogenic fungi and protozoa is the heart. Adult patients with hepatic hydatid disease complicated by paecilomycosis have been found to have atypical paecilomycosis-associated myocarditis, the treatment of which was developed by the authors, by using antibiotics, fungicides, and homeopathic remedies.

This study was performed for the purpose of investigating the prevalence of Sarcocystis spp. in buffaloes in Beni-Suef Governorate, Egypt. Both macroscopic (Sarcocystis fusiformis) and microscopic (Sarcocystis levinei) cysts were recognized, and were differentiated by their morphological features and location in the tissues. Of 379 buffaloes examined in abattoirs in Beni-Suef, 299 were found to be infected, with an overall prevalence of 78.9%. Depending on age, three categorized groups of naturally infected buffaloes were examined: male buffalo calves aged 1.5-2 years, adult females aged 2-5 years, and females older than 5 years. Among these groups, infection rates were 74.5%, 82.3%, and 81.2%, respectively. Organs examined included esophagus, tongue, and heart. Macroscopic cysts were examined by the naked eye through meat inspection in abattoirs, while the pepsin-digestion method and the histological technique were applied to detect microscopic cysts. It has been found that esophagus showed the highest rate of infection among the infected organs, with both macroscopic and microscopic cysts seen in the infected buffaloes. Moreover, results of the pepsin-digestion method proved more accurate than those produced by the histological technique in terms of infection rates for the microscopic cysts. Our findings indicated that infected buffaloes aged 2-5 years showed the highest mixed infection rate (82.3%) for both types of cysts. The high prevalence of microscopic Sarcocystis spp. in Beni-Suef Governorate reflects a significant role played by stray dogs, rather than cats, in the transmission of these parasites.

Controversy exists concerning whether cattle and water buffalo sustain infections with cysts of distinct arrays of species in the genus Sarcocystis. In particular, morphologically similar parasites have been alternately ascribed to Sarcocystis cruzi or to Sarcocystis levinei, depending on their occurrence in cattle or water buffalo. We used light and transmission electron microscopy, genetic analysis, and experimental infections of definitive canine hosts to determine whether consistent differences could be identified from parasites derived from several natural infections of each host, examining several tissue types (esophagus, skeletal muscles, and heart). Cysts derived from cattle and water buffalo shared similar structure; variation among 18S rRNA sequences did not segregate consistently according to intermediate host type; parasites derived from cattle and water buffalo induced similar outcomes in the canine definitive host. One cattle specimen harbored unusually large (macroscopic) sarcocysts which nonetheless conformed to previously reported ultrastructural and genetic features of S. cruzi. Finding no consistent basis to differentiate between them, we conclude that the parasites infecting each host and tissue type correspond to S. cruzi. In our sample, no phylogenetically distinct taxon was sampled which might correspond to a distinct taxon previously described as S. levinei. Either that taxon was missed by our sampling effort, or it may represent a junior synonym to S. cruzi, which would then cycle between dogs and a broader range of intermediate bovine hosts than was previously considered., (Copyright © 2010 Elsevier Inc. All rights reserved.)

This study was conducted to determine the prevalence, distribution pattern, and the Sarcocystis species involved in slaughtered water buffaloes (Bubalus bubalis) in the Khuzestan, Iran by macroscopic and histological examination. The esophagus, heart, diaphragm, tongue, masseter, and thigh muscles were investigated. Esophagus and thigh muscles of only 3 of the 100 examined water buffaloes (3%) were infected with macroscopic Sarcocystis, whereas at microscopic level Sarcocystis were found in 83 of the 100 examined animals (83%). The highest prevalence rate of microscopic cysts was found in masseter muscle (57.1%) and then followed by tongue, diaphragm, esophagus, heart, and finally, thigh muscles (30.0%). There was no significant difference between males (83.6%) and females (82.0%) or between two investigated age groups (2 years old, 88%). Based on the size of the sarcocysts, thickness of the walls and location of the cysts, Sarcocystis buffalonis as a macroscopic form and Sarcocystis levinei and Sarcocystis dubeyi as the microscopic forms were diagnosed in the examined muscles of the buffaloes of this area. Sarcocystis fusiformis was not seen in the examined organs of these buffaloes. The high prevalence rate of microscopic Sarcocystis in this region indicates that dogs have a more significant role than cats in transmission of these protozoa.

Sarcosporidian cysts in the skeletal muscle of domestic pigeons (Columba livia f. domestica) have previously been attributed to infection with Sarcocystis falcatula, which is shed in the faeces of the opossum (Didelphis virginiana). Here, we describe fatal spontaneous encephalitis and myositis associated with Sarcocystis infections in three flocks of racing pigeons with 47 of 244 animals affected. The clinical course was characterized by depression, mild diarrhoea, torticollis, opisthotonus, paralysis and trembling. Histopathological examination of 13 pigeons revealed generalized severe granulomatous and necrotizing meningoencephalitis and myositis with sarcosporidian cysts. Light and transmission electron microscopy identified cysts in heart and skeletal muscle of 1 to 2 mm in length and 20 to 50 microm in width. These were subdivided into small chambers by fine septae and filled with lancet-shaped cystozoites (7.5 x 1.5 microm) and dividing metrocytes, which is characteristic for Sarcocystis. The cysts had smooth walls and were devoid of protrusions typical of S. falcatula. Polymerase chain reaction amplification and sequencing of the internal transcribed spacer region (ITS-1) and the complete 28S rRNA identified a novel Sarcocystis species with only 51% ITS-1 nucleotide sequence similarity with S. falcatula. A phylogenetic comparison of the 28S rRNA revealed close sequence homologies with Frenkelia microti, Frenkelia glareoli and Sarcocystis neurona. The clinical, histopathological, electron microscopic and genetic data are unlike any previously described protozoan infections in pigeons, suggesting a novel, severe disease due to an as yet undescribed Sarcocystis species.

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.

Muscle tissues from 34 moose from Southeastern Norway and two moose from Canada were examined. Sarcocysts were excised and morphologically classified by light microscopy, and some cysts were further examined by scanning electron microscopy or DNA amplification and sequencing at the small subunit (ssu) rRNA gene. In Norwegian moose, three sarcocyst types were recognized, yet five Sarcocystis species were found by sequence analysis. New names were proposed for three species which could be characterised by both morphological and molecular methods, i.e., Sarcocystis alces, Sarcocystis ovalis, and Sarcocystis scandinavica. S. alces was the most prevalent species, whereas S. scandinavica and the two unnamed species were rare and might either use another principal intermediate host or a rare definitive host. The five species in Norwegian moose were different from Sarcocystis alceslatrans isolated from a Canadian moose. Phylogenetic analyses based on complete ssu rRNA gene sequences revealed a close relationship between the six Sarcocystis species from moose and species from reindeer and Sika deer. We conclude that molecular methods are necessary for unequivocal species identification, as different cervid hosts harbour morphologically indistinguishable sarcocysts.

Protozoal meningoencephalitis is considered to be an important cause of mortality in the California sea otter (Enhydra lutris). Thirty nine of 344 (11.3%) California (CA) and Washington state (WA) sea otters examined from 1985 to 2004 had histopathological evidence of significant protozoal meningoencephalitis. The aetiological agents and histopathological changes associated with these protozoal infections are described. The morphology of the actively multiplicative life stages of the organisms (tachyzoites for Toxoplasma gondii and merozoites for Sarcocystis neurona) and immunohistochemical labelling were used to identify infection with S. neurona (n=22, 56.4%), T. gondii (n=5, 12.8%) or dual infection with both organisms (n=12, 30.8%). Active S. neurona was present in all dual infections, while most had only the latent form of T. gondii. In S. neurona meningoencephalitis, multifocal to diffuse gliosis was widespread in grey matter and consistently present in the molecular layer of the cerebellum. In T. gondii meningoencephalitis, discrete foci of gliosis and malacia were more widely separated, sometimes incorporated pigment-laden macrophages and mineral, and were found predominantly in the cerebral cortex. Quiescent tissue cysts of T. gondii were considered to be incidental and not a cause of clinical disease and mortality. Protozoal meningoencephalitis was diagnosed more frequently in the expanding population of WA sea otters (10 of 31, 32.3%) than in the declining CA population (29 of 313, 9.3%). Among sea otters with protozoal meningoencephalitis, those that had displayed neurological signs prior to death had active S. neurona encephalitis, supporting the conclusion that S. neurona is the most significant protozoal pathogen in the central nervous system of sea otters.

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.

Transmission electron microscopy was used to the cysts and their walls of Sarcocystis arieticanis within cardiac muscle of naturally infected sheep. The general ultrastructural features of the cysts, previously described by other authors, were confirmed. The cyst walls of S. arieticanis and the size and shape of the protrusions at different locations on the cyst wall were explained in detail. The cysts were 35-62.5 micro m x 52.5-162.5 micro m in size and the cyst wall had different-shaped protrusions. Aged cysts that were localized in the heart and differences in the morphology of their protrusions were noticed.

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.

In January 1998 and 1999, two mass strandings of dolphins occurred in Wellfleet, Massachusetts. The strandings were composed of 97 and 53 animals, respectively. Tissues from 35 Atlantic white-sided dolphins (Lagenorhynchus acutus) from the 1998 stranding and 52 from the 1999 stranding were examined histologically. In the 1998 stranding, unidentified protozoal tissue cysts were seen in skeletal muscle from 11 of 28 (39%) dolphins. In addition, two dolphins had a protozoal tissue cyst in cardiac muscle. In the 1999 stranding, nine of 23 (39%) dolphins had the same protozoal tissue cysts in skeletal muscle. The identification of these protozoal tissue cysts as Sarcocystis sp. was confirmed by light and transmission electron microscopy. The high prevalence of sarcocysts in these dolphins suggests that they are likely intermediate hosts for previously undescribed Sarcocystis spp. The ultrastructure of the sarcocyst walls suggests that more than one species of Sarcocystis are present in dolphins.

Sarcocystis sp. schizonts were found in the cerebellum of a northern gannet (Morus bassanus), exhibiting neurologic signs, found on the Florida (USA) east coast. Based upon molecular characterization of DNA isolated from the brain of the gannet, this Sarcocystis sp. appeared to be closely related, if not identical, to an unnamed Sarcocystis sp. typified by isolates 1085 and 1086 collected from feces of a Virginia opossum (Didelphis virginiana) on the east coast of Florida. Because the life cycle of this parasite appears to be land based, urban waste discharge to marine/estuarine environments may be a source of infection for marine species.

Sarcocystis neurona associated granulomatous encephalitis was found in 2 of 84 adult raccoons. Both raccoons also had an extensive nonsuppurative myocarditis and one had S. neurona schizonts and merozoites in the myocardium. Only the asexual stages (schizonts and merozoites) of S. neurona are found in tissues of naturally infected animals (horse, mink, raccoons, cats, skunk, pony, seals, sea otters) and since these have not been reported outside the central nervous system, the presence of concurrent myocarditis in raccoons with the presence of S. neurona in the current study is of interest. Pathologists should consider the possible association of S. neurona with myocardial inflammation in other S. neurona susceptible animals.

Acute sarcocystosis was diagnosed in an adult female wild turkey (Meleagris gallopavo) that was collected from Early County (Georgia, USA) in February of 1998. Marked inflammatory lesions were seen in the heart, lung, and liver and were associated with protozoal schizonts and merozoites. The organisms were identified as Sarcocystis sp. (Acomplexa: Sarcocystidae) based on structure and antigenicity. Protozoa divided by endopolygeny, merozoites lacked rhoptries, and the organisms did not react to anti-S. falcatula antibodies but reacted to anti-S. cruzi antibodies.

Growth performance and the pattern of glycolytic enzymes in the blood plasma were assessed during experimental Sarcocystis cruzi infection (1 x 10(5) sporocysts per calf) in six calves; five calves served as noninfected controls. At slaughter (68 or 88 days post infection), carcass weight, dressing percentages and several parameters of meat quality (pH, color brightness, rigor, water absorbing capacity, water binding capacity) were recorded. Moreover, enzyme activities were measured in muscle homogenates. Weight gain was significantly impaired by the infection. Activities of lactate dehydrogenase (LDH) and aldolase (ALD) significantly increased in the blood plasma of the infected calves during the chronic stage of the disease, while glucose-6-phosphate dehydrogenase (G6PDH) and isocitrate dehydrogenase (ICDH) were not significantly altered. This was accompanied by a significant decrease of enzyme activities in the Musculus longissimus dorsi (LDH, ALD), in the diaphragmatic musculature (ALD, G6PDH) and in the heart (LDH, ALD). Activities of LDH, ALD, ICDH and G6PDH were visualized by enzyme histochemistry within the developing sarcosporidial cysts. However, isoenzymes of parasite origin could not be demonstrated by agar-gel electrophoresis of muscle homogenates or blood plasma. It is concluded that sarcocystiosis of even moderate severity alters the performance of calves but not meat quality. Leakage of glycolytic enzymes from the affected muscles is the probable cause of increased plasma enzyme activities. Although these enzymes are also synthesized by the parasite, the contribution of parasite-derived enzymes to the observed changes of enzyme patterns remains in question.

In a lifespan transgeneration study under standard laboratory conditions using a total of 4682 CBA/J mice, unusual intramuscular inclusions were found in the diaphragm, heart and skeletal muscle of one mouse using light microscopy. Located within the myocytes, they caused no visible tissue reaction. Cross-sections of these spherical and cystic lesions showed numerous banana-shaped structures, identified as permanent parasitic bradyzoites, which permitted these infections to be diagnosed microscopically as sarcocystosis.

Muscles from heart, tongue, oesophagus, neck and abdomen from 502 adult water buffaloes (Bubalus bubalis) slaughtered in Ho Chi Minh City, Vietnam, between 1996 and 1997 were examined for Sarcocystis cysts by a combination of ocular and histological examination. Sarcocysts were present in 396 (79%) of the animals and the prevalence increased with age from a 57% infection rate among 2-3 year old animals to 93% among 6-7 year olds. The prevalence was higher in animals originating from the northern part (89%) than in those from the southern part (69%) of the country. Four species of Sarcocystis were identified. S. levinei (74%) was the most common species found, followed by S. fusiformis (41%), S. buffalonis (33%) and S. dubeyi (12%). All four species were present in 8% of the infected animals. The most common site for sarcocyst location was oesophagus, followed by cervical muscles, tongue and heart.

An adult female hard-ground swamp deer (Barasingha), Cervus duvauceli branderi, was found dead of unknown cause in Kanha National Park. A necropsy of this animal failed to reveal any significant gross lesions. However, histopathologic examination revealed mature sarcocysts of Sarcocystis in the cardiac muscles. The morphology of the sarcocysts was similar to that of different species of Sarcocystis from wild and domestic animals. Final identification of Sarcocystis in this case could not be made. This is the first recorded case of Sarcocystis infection in hard-ground Barasingha deer.

The prevalence of Sarcocystis spp. infection was investigated in 605 sheep, 826 goats, 1080 cattle, 580 water buffaloes and 36 camels slaughtered from 1992 to 1996 in the Baghdad area (Iraq) using naked eye examination for macroscopic sarcocysts, and peptic digestion, muscle squash, squeezing methods and indirect fluorescent antibody test (IFAT) for microscopic types. The intestinal stages of the parasite were also studied in dogs experimentally fed with tissues containing microscopic cysts. The percentage prevalence of macroscopic cysts were 4.1, 33.6, 0.2, 15.6 and 0, and of the microscopic type, 97.0, 97.4, 97.8, 82.9 and 91.6 for the above-mentioned hosts, respectively. Among the different organs examined, macroscopic cysts were found to be highest in the oesophagus and the lowest in the heart. Peptic digestion method gave the highest rate (93.3%) followed by indirect fluorescent antibody test (IFAT) (88.6%), squeezing (81.3%), and muscle squash (81.2%). Each infected dog shed a total of about 150-200 million sporocysts. Histologically, developmental stages of the parasite were detected in the small intestinal mucosa of the dogs on Days 7 and 13 post-infection.

A captive-born juvenile female rhesus macaque (Macaca mulatta) was acquired from a commercial breeder and placed in quarantine. Within 8 days of arrival, the animal became anorexic, inactive, and dehydrated. Subsequently, generalized edema and facial ecchymoses developed, and despite supportive therapy, the animal became moribund and was euthanatized. Macroscopic examination showed diffuse stippling and streaking of the myocardium. Histopathologic examination revealed multifocal to coalescing myocardial edema, necrosis, lymphohistiocytic inflammation, and generalized endothelial infection with Sarcocystis sp. Immature and mature schizonts within endothelial cells were most prevalent in the heart. Fewer schizonts were present in the vasculature of other tissues, including skeletal muscle, smooth muscle, adipose tissue, brain, and retina. Mature tissue cysts within muscle fibers were not found in the myocardium but were occasionally seen in skeletal muscle. Analysis of polymerase-chain-reaction-amplified 18s ribosomal RNA gene sequences revealed 96% identity to published sequences of S. hirsuta, S. hominis, and S. fusiformis and 95% identity to S. cruzi and S. tenella. However, sequences did not show complete identity with any organism in the GenBank database. Sequence homology suggests that this is a newly described Sarcocystis sp. Results of antibody tests for simian retrovirus, simian T-lymphotropic virus 1, and simian immunodeficiency virus were negative, suggesting that viral immunosuppression was unlikely to have augmented the pathogenicity of sarcosporidial infection. Clinical and histopathologic findings in this case of fulminant sarcosporidiosis are similar to those described in Dalmeny disease in cattle, which is associated with ingestion of massive numbers of infective Sarcocystis oocysts.

A Sarcocystis-like organism was associated with encephalitis and myocarditis in an ataxic, emaciated adult male turkey (Meleagris gallopavo) from Gilmer County, West Virginia. Protozoal schizonts and merozoites were associated with areas of inflammation and occasionally necrosis in both the heart and the brain. The organisms divided by endopolygeny and stained positively with anti-Sarcocystis cruzi serum in an immunohistochemical test.

Sarcoystis suihominis was detected for the first time in Japan from the heart and diaphragm of 5 out 600 older culled breeding pigs slaughtered in Saitama Prefecture, Japan. Fresh cysts were 1,080-2,040 x 106-170 microns in size. Bradysoites measured 15 x 4 microns on average. The cyst wall was usually observed thick, 4-6 microns, and striated, but occasionally thin and smooth according to the difference in sectioning angle and in portion of cysts. Scanning electron microscopy showed that many palisade-like villar protrusions, 6-6 x 0.3-0.5 microns in size, were closely folded onto the surface of cyst. A small number of microtubules were seen in the core of protrusion. No dogs nor domestic cats fed with 20 fresh cysts each excreted oocysts or sporocysts in the feces throughout the experimental period of 30 days.

Acute generalized sarcocystosis was diagnosed in a capercaillie (Tetrao urogallus) from Finland. Microscopic lesions were seen in the heart, lungs, spleen, liver, and brain. Protozoa were found in all organs, especially in the lungs and spleen. Only asexual stages were observed. The parasite divided by endopolygeny. Schizonts were usually 10 microm wide and up to 55 microm long. Merozoites are 3-4 microm long and 1.5-2.0 microm wide. Sarcocysts and sexual stages are unknown. The parasite was considered to be a species of Sarcocystis with an unknown life cycle. This is the first report of acute sarcocystosis in capercaillie from Finland.

Sarcocystis levinei Dissanaike and Kan, 1978, is redescribed because the original description was a mixture of 2 species, the amended S. levinei and the newly recognized S. buffalonis Huong, Dubey, Nikkilä, and Uggla, 1997. In histological sections, S. levinei sarcocysts are microscopic, up to 640 microm long and up to 95 microm wide. Ultrastructurally, the cyst wall is thin (< 1.0 microm thick) with a minute undulating surface and smooth, hairlike villar protrusions arising at irregular distances from the cyst wall. The villar protrusions have a dome-shaped base (approximately 0.5 microm thick), a fingerlike middle part, and a tapering distal end (<0.1 microm thick). The morphological features of the sarcocyst resemble those of S. cruzi (Hasselman, 1926) Wenyon, 1926, of cattle. Sarcocystis levinei sarcocysts were found in striated muscles including heart, esophagus, tongue, and skeletal muscle. The buffalo myocardium is parasitized exclusively by S. levinei, whereas >1 Sarcocystis species may occur concurrently in other muscular tissues of water buffaloes. Two dogs, but not 2 cats, fed water buffalo hearts infected with S. levinei sarcocysts shed sporocysts measuring 9.5-10.5 x 14.0-16.5 microm starting from days 16 and 18, respectively.

Samples of heart, tongue, oesophagus and diaphragm muscle from twenty-two naturally infected roe deer (Capreolus capreolus) harvested in central Italy were examined for sarcosporidiasis. The structure of Sarcocystis spp. muscle cysts was examined by light and electron microscopy. Only one type of thin-walled cyst was distinguished by light microscopy. Electron microscopy showed cysts having a thin highly folded primary cyst wall, without fibrillar material, that formed thin hair-like protrusions often having a T-form, especially close to host cell mithocondria. The cysts appeared to belong to a single Sarcocystis sp. so that all the animals had monospecific infections. This cyst was compared with cysts described in other cervid in an attempt to determine if single or multiple species of the genus Sarcocystis occur in the Cervidae. Apparently, a single Sarcocystis sp. with a low specificity for the intermediate host can infect the Cervidae.

Collagen cross-reactive antigenic substance(s) in Sarcocystis cruzi cysts were examined with immunologic techniques using anti-bovine collagen type-specific, but non-species-specific, antibodies. By immunoperoxidase test, anti-bovine collagen type-specific, but non-species-specific, antibodies. By immunoperoxidase test, anti-bovine type IV collagen antibody showed higher reactivity to the cysts than other antibodies tested. Cyst wall rupture was induced by collagenase treatment and digestion was inhibited with EDTA supplementation. With immunoblotting analysis, one band of the cyst extract, which exhibited specific reactivity to anti-bovine type IV collagen antibody, was detected. The band had a molecular weight of approximately 66 kDa. These results suggest that sarcocysts of S. cruzi may be comprised of bovine collagen type IV cross-reactive antigenic substances.

The oesophagus, diaphragm and heart of 103 camels slaughtered at Al-Ahsa abattoir were investigated for infection with sarcocysts. Sarcocystis cysts were found in 91 camels (88.35%): in the diaphragm of 79.6%, oesophagus of 72.8% and heart of 71.8%. Two morphologically distinct sarcocysts were found. Thin-walled cysts (141-400 x 70.5-188 microns) were in all the three indicator organs while the thick-walled cysts (170-194 x 117.5-188 microns) were only found in the oesophagus. Experimental feeding of four cats and two dogs with sarcocysts-infected meat showed that cats were refractory to the infection while dogs excreted Sarcocystis sp. sporocysts (10.7-14.3 x 8.3-10.7 microns mean, 13.2 +/- 0.32 x 9.4 +/- 0.20 microns) after a prepatent period of 9-10 days. It was concluded that dogs may be the final host of both types of sarcocysts or that one of these cysts develops in dogs while the other one uses other carnivores living in the desert as the final host.

A survey of Sarcocystis was conducted in cattle, sheep, goats, donkeys and chickens. A total of 671 haematoxylin-eosin (H-E) stained muscle tissue samples, including diaphragm, masseter, cardiac and oesophageal musculatures were examined. Additionally, cardiac muscle samples from 40 fetuses were included. An infestation rate of 93% in sheep, 82% in cattle, 81% in goats, 16.6% in donkeys and 6.6% in chickens was noted. The infestation rate of diaphragm, masseter, cardiac and oesophageal musculatures seems to be similar. None of the 40 fetal heart muscle samples from bovine, ovine, caprine and donkey fetuses examined harboured Sarcocystis. An attempt was made to demonstrate the possible occurrence of human Sarcocystis and a negative result was obtained. The possible impact of Sarcocystis on animal health in Ethiopia is discussed.

Four 4-mo-old elk calves (Cervus elaphus) obtained from northeastern Oregon (USA) each were inoculated orally with 250,000 sporocysts of Sarcocystis spp., including S. sybillensis and S. wapiti. Three similar elk calves of comparable ages and weights served as uninoculated controls maintained with the inoculated elk during the experimental period between September and December 1993. Body weights were evaluated at 0 and 90 days postinoculation (PI); packed cell volumes of whole blood were evaluated at 0, 30, and 60 days PI, and numbers of sarcocysts in histologic sections from 11 selected tissues were evaluated at 90 days PI. Significant differences in blood packed cell volumes were not detected between groups (P > 0.05). Except for weight gain, elk remained healthy. Mean (+/- SE) weight gain of inoculated elk (27.1 +/- 1.6 kg) was significantly (P < 0.05) less than that of controls (40.2 +/- 4.9 kg). Mean (+/- SE) number of sarcocysts in tissues of inoculated (114.4 +/- 25.7 cm2) and controls (4.5 +/- 1.4 cm2) differed significantly (P < 0.05). Heart, esophagus and skeletal muscle contained the most sarcocysts. No sarcocysts were detected in brain, spinal cord, or testicles. Histologically, mononuclear myositis and myocarditis, with numerous intralesional sarcocysts were seen. Less severe, but widespread inflammation occurred in brain, spinal cord, and optic nerve. Mortality and anemia were not seen, but weight gain depression was detected in the inoculated elk over the 90 day experimental period.

Three species of sarcocysts from European roe deer (Capreolus capreolus) are characterised using light and electron microscopy. The hitherto existing descriptions of four species are attributed to the actual state of nomenclature.

The polymerase chain reaction (PCR) method to randomly amplify polymorphic DNA (RAPD) was used to differentiate between Sarcocystis cruzi DNA and bovine DNA. This assay was also exploited to identify a S. cruzi DNA fragment which may be useful as a probe. Five primers ranging in length from 16 to 20 nucleotides were analyzed for their ability to direct the amplification of either bovine or parasite DNA fragments. Two primers, TGA and TGD, preferentially amplified bovine DNA in a mixture of S. cruzi and bovine DNA. The primers TGB and TGF each directed the amplification of S. cruzi DNA instead of bovine DNA. Assays using TGF and S. cruzi DNA resulted in the production of a unique 0.8 kilobase (kb) DNA fragment. This fragment was not amplified from two other closely related coccidian species, Toxoplasma gondii and Sarcocystis campestris. When the 0.8 kb DNA fragment was purified and used as a DNA probe, it only hybridized with DNA from S. cruzi. The results of this study indicate that this DNA fragment may be developed into a useful DNA probe for S. cruzi, and that the RAPD-PCR method may be successfully exploited for the rapid development of DNA probes for parasites and other organisms.