The distribution pattern of Sarcocystis species,their transmission and pathogenesis in sheep in Fars Province of Iran

Of 1362 sheep examined during two years in Fars Province of Iran, 786 (57.7%) were positive for Sarcocystis spp. The prevalence was significantly higher (p<0.05) in animals owned by nomadic Assyrians (67.95%) than in those owned by local people (41.86%). More of the animals above 2 years age were infected (69.98%) than young ones (30.02%). Females had a higher prevalence of infection (61.07%) than males (38.93%) but most of the males were younger. There was no variation in the infection rate during spring, summer or autumn, but it was low in winter. The species observed were Sarcocystis gigantea, predominantly in oesophagus, S. medusiformis, mainly in diaphragm, S. tenella in the oesophagus, diaphragm, tongue and heart, and S. arieticanis in the oesophagus, tongue and occasionally in the diaphragm. In transmission studies, the prepatent period for S. gigantea and S. medusiformis and for the two microscopic species was 11–13, 10 and 8–12 days, respectively. The infection could not be transmitted to hamsters and guinea-pigs. The macroscopic species were almost non-pathogenic but were responsible for economic losses because of rejection of carcases or parts thereof at slaughter. The microscopic species caused tissue damage to the affected organs, resulting in haemorrhages, mononuclear infiltration and necrotic changes.Abbreviations DPI days post infection     

Morphological and molecular identification of three species of Sarcocystis in reindeer (Rangifer tarandus tarandus) in Iceland

Six Sarcocystis species have previously been described from reindeer in Norway based on sarcocyst morphology and DNA sequencing. The aim of this study was to determine whether reindeer in Iceland, which descend from reindeer imported from Norway in 1787, also were infected with Sarcocystis, and to identify and genetically characterise any species present. Muscle tissue from the heart, diaphragm and/or oesophagus was collected from 36 reindeer in Iceland. Pieces of all tissue samples were examined histologically. Frozen/thawed samples of cardiac muscle, oesophagus and/or diaphragm from 11 of the 36 reindeer were also examined under a stereoscopic microscope and sarcocysts present were identified to species either in situ or under a light microscope. Two cysts of each species, originating from two different reindeer were randomly selected for DNA analyses. The complete ssu rRNA gene was amplified by the polymerase chain reaction (PCR) and sequenced. In addition, two sarcocysts that could not be classified by microscopic examination were selected for partial ssu rRNA gene sequence analysis. By histology, sarcocysts were found in the diaphragm and/or oesophagus of 8 of 36 (22.2%) animals. By examination of fresh tissue, sarcocysts of Sarcocystis rangi, S. tarandivulpes and S. hardangeri were found in the oesophagus of seven of nine (77.8%) animals, suggesting a high prevalence of Sarcocystis in the Icelandic reindeer population. Cyst morphology and the ssu rRNA gene sequence of each of the three species were identical to isolates of the same species from Norwegian reindeer. DNA sequencing was useful in order to identify cysts with an ambiguous morphology. This is the first record of these Sarcocystis species in reindeer outside Norway.     

Sarcocystis infections in gazelles at the King Khalid Wildlife Research Centre, Saudi Arabia

The prevalence of Sarcocystis species in muscle samples from gazelles kept as breeding groups at the King Khalid Wildlife Research Centre, Saudi Arabia, was determined by fibreoptic examination, pepsin digestion and histological techniques. No macroscopic sarcocysts were detected by fibreoptic examination, and the overall prevalence of Sarcocystis was 66 x 7 per cent by pepsin digestion, and 39 x 9 per cent by histological examination. By digestion, the tongue contained the highest density of bradyzoites in Gazella dorcas, and Gazella gazella erlangeri, the oesophagus in Gazella subgutturosa marica and skeletal muscle in Gazella gazella and Gazella thomsoni. Skeletal muscle was least affected in G dorcas, the oesophagus in G gazella, and the diaphragm in G g erlangeri, G s marica and G thomsoni. By histology, the heart contained most microcysts, except in G g erlangeri, in which the tongue was most affected. No single tissue type was therefore suitable for the diagnosis of sarcocystosis in this multispecies collection, although digestion was more sensitive in detecting infection than histology. The level of Sarcocystis infection was significantly higher in free-ranging gazelles kept in a main enclosure than in gazellas kept in breeding pens, and higher in adult gazelles than in juveniles.     

The prevalence and identity of Sarcocystis cysts in cattle in Belgium

Muscle tissue from the oesophagus, diaphragm and heart of 100 cattle slaughtered in Belgium was examined for Sarcocystis infection by microscopic examination of tissue and artificial digestion. Intact sarcocysts or cystozoietes were recovered from 97% of the cattle examined. There was a difference in sensitivity between the method (digestion or histology) used and the muscle processed. The digestion of the oesophagus muscle resulted in the highest number of positive animals whereas the heart muscles contained most cysts during histological examination. Thin-walled cysts were recovered from all positive animals especially in the heart and they were indistinguishable from those of S. cruzi. Thick-walled cysts were recovered from 56% of animals but these could not be identified as S. hirsuta and/or S. hominis on morphological grounds. A correlation between pathological changes and the infection grade could not be proved.     

Prevalence of Sarcocystis sarcocysts in nine-banded armadillos (Dasypus novemcinctus) from Florida.

The prevalence and identity of Sarcocystis spp. sarcocysts in the skeletal muscles of nine-banded armadillos (Dasypus novemcinctus) collected from Alachua County, FL, were determined. H & E stained sections of skeletal muscle from tongue and thigh were examined. Thirty nine of 63 (61.9%) armadillos examined contained Sarcocystis sarcocysts. Two species were identified, Sarcocystis dasypi and Sarcocystis diminuta. Sarcocystis dasypi sarcocysts were found in 38 of 63 (60.3%) and S. diminuta sarcocysts were found in 6 of 63 (9.5%). Sarcocysts of S. dasypi were larger, more densely packed with bradyzoites, and bradyzoites contained within the sarcocyst were smaller than those of S. diminuta. Mixed infections occurred in 5 of 63 (7.9%) armadillos examined.     

Prevalence,biology, and distribution pattern of Sarcocystis infection in water buffalo (Bubalus bubalis) in Iran

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, 78% versus >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.     

Prevalence of sarcocysts of Sarcocystis capracanis in oesophagus and tail muscles of naturally infected goats

The prevalence of sarcocysts of Sarcocystis capracanis in oesophagus and tail muscles of 76 goats slaughtered at Jabalpur, India, was found to be 67.10 and 46.05%, respectively. A total of 79.3% of oesophagi and 72.4% of tails had sarcocysts in older goats (greater than 1 year), but their prevalence in younger animals (less than 1 year) was 40.0% in oesophagi and none in tails. Therefore examination by biopsy of the tail muscles can be used to monitor the development of Sarcocystis sarcocysts in experimental infection without killing the animal.     

Prevalence of Sarcocystis spp. in Argentinean cattle

Sarcocystis cruzi, S. hirsuta and S. hominis are apicomplexan parasites that affect cattle worldwide with variable prevalence. The aim of the present study was to evaluate the prevalence of Sarcocystis spp. in Argentinean cattle comparing microscopic fresh examination and molecular methods. Blood, myocardium and loin samples were collected in five slaughterhouses from a total of 380 bovines. Origin of animals was representative of the major beef cattle production area of Argentina. Samples were analyzed by fresh microscopical examination, transmission electron microscopy (TEM), IFAT and PCR-RFLP. Thin walled sarcocysts corresponding with S. cruzi were found in 99.5% of heart samples. Sarcocysts were detected in 73.1% of loin samples; 71.5% had S. cruzi cysts and 23.1% had thick walled sarcocysts (S. hirsuta or S. hominis). TEM observation revealed the presence of characteristic S. hominis and S. hirsuta cyst walls in 7 and 1 loin samples respectively. Using IFAT, 379/380 animals had titers 25 or higher, showing a full agreement with fresh examination. Amplification products were detected in 35.5% (135/380) of loin samples; however Sarcocystis species could only be determined by RFLP in 29 samples. Agreement between fresh examination and PCR was low (Kappa value=0.262). This is the first report of S. hominis and S. hirsuta in Argentina. Further studies are needed to improve the sensitivity of molecular methods for species identification, especially for differentiation of S. cruzi and S. hirsuta from the zoonotic species S. hominis. The results of the present study and others focusing on sensitivity and specificity of Sarcocystis spp. diagnostic methods should contribute to improve food safety.     

Prevalence of Sarcocystis species in sheep and goats in northern Nigeria.

Tissue samples comprising the oesophagus and diaphragm were collected from 400 sheep and 400 goats slaughtered at the abattoirs in the study area. Out of this number, 36 were positive for Sarcocystis cysts (sarcocysts) in sheep and 56 in goats. The sarcocysts in sheep measured 35.7 to 500 microns lengthwise and the cyst-wall 2.4 microns. They were identified to be Sarcocystis tenella. The cysts in goats measured 98 to 700 microns and the cyst-wall 2.7 microns. They were identified to be Sarcocystis capracanis. In both animals species, the sarcocysts were more frequent in the oesophagus than in the diaphragm. All sarcocysts seen were microscopic.     

Sarcocystis Infection in Sheep from South-Western Norway

The occurrence of Sarcocystis infection and pathological changes were recorded in samples of the heart, diaphragm, and oesophagus from 198 healthy sheep representing 3 different age groups, obtained from an abattoir.The infection rate of S. gigantea (syn. S. tenella) was 18.2 %, and the distribution within groups was: ewes 30.0 %, yearlings 11.6 %, lambs nil. The infection rate of S. tenella (syn. S. ovicanis) was 65.1 %, and the corresponding distribution was: ewes 83.5 %, yearlings 74.4 %, and lambs 25.0 %. A third type of Sarcocystis sp. displaying thick wall was found in 3 samples.Focal interstitial infiltrates of mononuclear cells were demonstrated in 47.9 % of the hearts, in 19.6 % of the diaphragms and in 31.3 % of the oesophagi. The occurrence of Sarcocystis and the focal interstitial mononuclear cell infiltrates were positively correlated (P < 0.0001). Morphologically identical sporocysts typical of S. tenella were produced by dogs and foxes fed naturally infected sheep tissues. A cat fed S. gigantea macrocysts produced sporocysts characteristic for the species.Sarcocystis; pathology; life cycle; final hosts; sheep.     

Prevalence of Sarcocystis spp. in meat-producing animals in Iraq.

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.     

Intramuscular inoculation of cattle with Sarcocystis antigen results in focal eosinophilic myositis

Bovine eosinophilic myositis (BEM) is a subclinical myopathy characterized by multifocal white to grey-green discolorations in skeletal muscles, heart, tongue and oesophagus. These lesions are found at slaughter or during meat cutting and result in considerable economic losses. The etiology and pathogenesis are unclear, although it has been suggested, that Sarcocystis species are involved. To elucidate their role, two calves were repeatedly injected intramuscularly with adjuvanted Sarcocystis antigen. The morphological changes at the injection sites in these calves were histologically and immunohistochemically compared to spontaneous lesions from 44 BEM condemned carcasses sampled in slaughterhouses. Experimental intramuscular injection of Sarcocystis antigen resulted in lesions at the injection sites that were similar to the lesions of natural cases of BEM. They were characterized by massive infiltration of eosinophilic granulocytes, reactive macrophages (MAC387(+) cells), T-cells (CD3(+)) and B-cells (CD20(+)). Both in the experimental and in the natural cases, COX-2 expression was present in endothelial cells adjacent to lesional areas. MHC class II(+) staining was found amongst others in muscle cells surrounding the lesion. These results show that Sarcocystis antigens can induce an inflammatory response in bovine muscle having the characteristics of natural BEM.     

Prevalence of Sarcocystis in domestic pigs in India.

Muscle samples from 890 slaughtered pigs were examined for the presence of sarcocysts. A high prevalence rate of 67.98% was observed. Two types of microsarcocysts were recorded. The sarcocyst wall of one type had redial striations and the other possessed hair-like villar protrusions. The species were identified as Sarcocystis miescheriana and Sarcocystis suihominis; there was a slightly higher incidence of the latter species (47.11%) than of the former (43.14%). S. suihominis has been identified for the first time from pigs in India.     

The fox as definitive host for Sarcocystis sp. Gjerde, 1984 from skeletal muscle of reindeer (Rangifer tarandus). With a proposal for Sarcocystis tarandivulpes n. sp. as replacement name

Skeletal muscle of 5 wild reindeer was examined for sarcocysts and used for experimental infection of 6 foxes. Skeletal and cardiac muscle of another reindeer were only examined for sarcocysts. The skeletal muscle of all animals was infected with Sarcocystis sp.. In 2 of the animals cysts of S. hardangeri were also present. The single heart examined contained only cysts of S. grueneri.Four foxes given skeletal muscle containing apparently only cysts of Sarcocystis sp., started shedding Sarcocystis sporocysts, measuring on average 13.6×9.8 µm, after a prepatent period of 10–12 days. Two foxes given skeletal muscle containing cysts of both Sarcocystis sp. and S. hardangeri shed similar sporocysts, measuring on average 13.5×9.7 µm, after a prepatent period of 10–12 days.Based on the results from the present and previous investigations, Sarcocystis sp. is considered to have foxes (Vulpes vulpes and Alopex lagopus) and dogs (Ganis familiaris) as definitive hosts, becoming the second species of Sarcocystis with a known reindeer/Canidae life cycle. The name Sarcocystis tarandivulpes n. sp. is proposed as a replacement name for Sarcocystis sp. Gjerde, 1984 from skeletal muscle of reindeer.     

The prevalence and identity of Sarcocystis in beef cattle in New Zealand

Muscle tissue from the oesophagus and diaphragm of 500 beef cattle slaughtered in New Zealand was examined for Sarcocystis infection by microscopic examination of cysts isolated from muscle samples. All cattle were infected with Sarcocystis; based on light microscopy of cysts, 98% had thin-walled Sarcocystis cruzi cysts and 79.8% had thick-walled (Sarcocystis hirsuta/Sarcocystis hominis) cysts. Cysts were also collected for electron microscopy and transmission experiments. Thick-walled cysts could not be distinguished as S. hirsuta or S. hominis by light or electron microscopy. Thick-walled cysts were fed to three cats and one human volunteer; one cat shed sporocysts but not the human volunteer. Electron microscopy of the cysts revealed many features that have not been described previously.     

Phylogenetic relationships between Sarcocystis species from reindeer and other Sarcocystidae deduced from ssu rRNA gene sequences

Six Sarcocystis species from reindeer (S. grueneri, S. rangi, S. tarandivulpes, S. hardangeri, S. rangiferi and S. tarandi) have previously been genetically characterised. The aim of this study was to identify possible definitive hosts for S. hardangeri, S. rangiferi and S. tarandi by including the six species in phylogenetic analyses of the Sarcocystidae, and also to investigate the phylogenetic relationships between the species from reindeer and those from other hosts. The study also aimed at revealing whether the inclusion of six Sarcocystis species from the same intermediate host would have any effect on previously inferred phylogenetic relationships within the Sarcocystidae. The complete small subunit (ssu) rRNA gene sequences of all six Sarcocystis species from reindeer were used in the phylogenetic analyses along with ssu rRNA gene sequences of 85 other members of the Coccidea. Trees were constructed using Bayesian analysis and maximum likelihood estimations. All six Sarcocystis species from reindeer were placed together with other Sarcocystis species using an even-toed ungulate as their intermediate host. The three canine transmitted species, S. grueneri, S. rangi, S. tarandivulpes, formed a sister group to other Sarcocystis species with a canine definitive host. The position of S. hardangeri on the tree suggested that it uses another type of definitive host than the other Sarcocystis species in this clade. Considering the geographical distribution and infection intensity of S. hardangeri, corvid birds are perhaps its most likely definitive hosts. The phylogenetic position, geographical distribution, prevalence and morphological similarity to feline transmitted Sarcocystis species in closely related Cervidae suggest that the most likely definitive hosts of S. rangiferi and S. tarandi are felines, and in Norway notably the lynx. The overall phylogeny of the Sarcocystidae did not change by the inclusion of the six Sarcocystis species from reindeer. This study suggests that phylogentic analysis can be a useful tool in the search for possible definitive hosts for those Sarcocystis species for which they are unknown and difficult to find solely by other methods.     

Sarcocystis capracanis and Toxoplasma gondii infections in range goats from Texas

Specimens of tongues, esophagi, diaphragms, or abdominal muscles of 115 range goats from San Angelo, Tex, were examined for Sarcocystis and Toxoplasma gondii infections. Sarcocystis spp zoites were found microscopically in pepsin digests of muscles of 60.8% goats and sarcocysts of S capracanis were found in histologic sections of 27.8% goats. Sarcocysts were more common in sections of tongue (19.1%) than in those of other muscles (9.9% to 10.7%). A dog fed Sarcocystis-infected tissues shed sporocysts in feces, whereas 2 cats fed the same tissues did not shed sporocysts. Toxoplasma gondii was neither seen in histologic sections of goat tissues nor found by bioassays in mice or cats. Mice inoculated with pepsin digests of muscles did not develop T gondii infection and 2 cats fed goat tissues did not shed oocysts. Also, antibody to T gondii was not found in serum samples from goats. The low prevalence of T gondii infection in range goats may be because of the relative absence of domestic cats on Texas ranges.     

Vigilance for veterinary medicinal products: declarations of adverse reactions in the year 2009

During the year 2009, 134 reports of suspected adverse drug reactions (ADRs) to veterinary medicinal products (VMPs) were received (106 in the year 2008). The distribution according to species and drug classes remained in line with previous years. Companion animals were involved in most of the reports (46 % dogs, 19 % cats), followed by cattle or calves (22 %). Antiparasitic drugs made the biggest part with 30 % of the reports, followed by antiinfectives (19 %) and hormones (13 %). Some reactions following their use are specifically discussed. 95 additional enquiries about ADRs of VMPs were received by the Swiss Toxicological Information Centre in Zürich. Most of them concerned dogs or cats and antiparasitics or anti-inflammatory drugs. In the vaccinovigilance program, a total of 1020 reports were received, of which 1000 were related to the vaccination against blue tongue disease. The most frequently reported adverse reactions were aborts, mastitis or alterations of milk quality and they are specifically discussed.     

Sarcocystis infections in Georgia swine

Tissues from 168 mature sows obtained at slaughter in northern and southern Georgia were examined for infection with Sarcocystis spp. Digestion techniques revealed zoites in 28 (16.5%) sows. Infected meat was fed to laboratory-reared dogs, cats, raccoons (Procyon lotor), and opossums (Didelphis marsupialis). Dogs and raccoons shed sporocysts (8.3 mum X 11.2 mum) 12 to 14 days after infection. Cats and opossums were refractory to infection. Sarcocystis-free pigs were infected with 50,000 sporocysts of swine-dog origin. Tissues from laboratory-infected swine were fed to dogs and raccoons. Both species shed sporocysts 14 days later. This is the 1st time in which a definitive host has been demonstrated for species of Sarcocystis occurring in North American swine. The raccoon constitutes a new definitive host for S suicanis Erber 1977. In contrast to previously reported low prevalences of Sarcocystis infections in swine, the relatively high prevalence reported here indicates that S suicanis may be of importance to swine producers in Georgia.     

Prevalence of Yersinia species in goat flocks

AIMS: To investigate the epidemiology of Yersinia species in healthy goats in New Zealand, in particular to determine the prevalence of farms with infected goats, the prevalence of infected goats on those farms, the serotypes involved, and potential risk factors for carriage. METHODS: A cross-sectional study of the prevalence of Yersinia infection in infected flocks in a study population of thirty commercial goat farms in the Manawatu region of New Zealand. RESULTS: Infection was detected on 60% of farms in an initial study. In a prevalence study on 18 infected farms, the study population comprised 6770 animals (mean of 376, median of 175 and range of 36 to 1295 goats/farm). Of 902 goats (296 < 1 year, 178 1 to 2 years, and 428 > 2 years) sampled from the study population, 135 (73 < 1 year, 21 1 to 2 years, and 41 > 2 years) were excreting Yersinia spp, giving an overall prevalence of 14.97% (95% confidence interval [CI]; 12.8 to 17.4) with individual farm prevalences ranging from 0.0 (+ 7.9) to 58.14% (95% Cl, 43.3 to 71.6). Goats < 1 year were more likely to be infected than 1-2 year and > 2 year old animals (relative risk [RR] = 2.1; 95% Cl, 1.3 to 3.3) and 2.6 (95% Cl, 1.8 to 3.6) respectively), but there was no significant difference between risks for 1 to 2 year and > 2 year goats (RR = 1.2; 95% CI, 0.7 to 2.0). Yersinia enterocolitica was the most common species isolated in the youngest age group, with prevalence declining with increasing age, while other species were more common in the older age groups. CONCLUSION: Yersinia infections were common in goats in the study region, with younger animals apparently more susceptible to infection and in particular to infection with Y enterocolitica. The prevalence on infected farms appeared to decrease as flock size increased and to increase as stocking rates and the number of paddocks grazed increased.