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Sample records for sarcocystis greineri sarcocysts

  1. Ultrastructure of Sarcocystis bertrami sarcocysts from naturally infected donkey (Equus asinus) from Egypt

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    There is considerable confusion concerning Sarcocystis species in equids. Little is known of Sarcocystis infections in donkeys (Equus asinus). Here we describe the structure of Sarcocystis bertrami-like from the donkey by light and transmission electron microscopy (LM, TEM). Nineteen sarcocysts fro...

  2. Sarcocysts of an unidentified species of Sarcocystis in the sea otter (Enhydra lutris)

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    Dubey, J.P.; Lindsay, D.S.; Rosenthal, B.M.; Thomas, N.J.

    2003-01-01

    The number of Sarcocystis species that infect sea otters (Enhydra lutris) is unknown. Sea otter tissues were recently shown to harbor sarcocysts of S. neurona and of unidentified species of Sarcocystis. Whereas sarcocysts of S. neurona have walls 1a??3 I?m thick with type 9 villar protrusions, ultrastructure of a distinct thin-walled sarcocyst (0.5a??0.7 I?m thick) lacking villar protrusions, but instead exhibiting minute type 1 undulations on the sarcocyst wall, is described in this report. Parasites characterized from a sea otter infection were inferred to be related to, but distinct from, other species belonging to Sarcocystis, based on sequencing and phylogenetic analysis of a portion of the beta subunit of the plastid-encoded RNA polymerase gene.

  3. Morphological and molecular characterization of Sarcocystis arctica-like sarcocysts from the Arctic fox (Vulpes lagopus) from Alaska, USA.

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    Cerqueira-Cézar, Camila K; Thompson, Peter C; Verma, Shiv Kumar; Mowery, Joseph; Calero-Bernal, Rafael; Antunes Murata, Fernando H; Sinnett, David R; Van Hemert, Caroline; Rosenthal, Benjamin M; Dubey, Jitender P

    2017-07-01

    The muscles of herbivores commonly harbor sarcocysts of parasites belonging to species in the genus Sarcocystis, but such muscle parasites are rare in carnivores. Here, we report Sarcocystis arctica-like sarcocysts in muscles of Arctic foxes (Vulpes lagopus) from Alaska, USA, for the first time. The tongues of 56 foxes were examined for Sarcocystis infection using several methods. Sarcocystis bradyzoites were detected in pepsin digests of 13 (23.2%), and sarcocysts were found in histological sections stained with hematoxylin and eosin (HE) of 9 (16.0%). By light microscopy, sarcocysts were up to 4 mm long and up to 245 μm wide. In HE-stained sections, the sarcocyst wall appeared smooth and up to 1.5 μm thick without visible protrusions. By transmission electron microscopy, the sarcocyst wall had a wavy parasitophorous vacuolar membrane (pvm) folded as pleomorphic villar protrusions (vp), sometimes with anastomoses of villar tips. The vp and the ground substance (gs) layer were smooth and without microtubules. The gs was up to 2.0 μm thick. The total width of the wall including vp and the gs was up to 4.0 μm. The vp were up to 3.0 μm long and most closely resembled "type 9c." All sarcocysts were mature and contained numerous 8.1 × 2.1 μm sized bradyzoites. Molecular characterization (at 18S rDNA, 28S rDNA, ITS-1, and cox1) showed the highest affinity for S. arctica of the Arctic fox (V. lagopus) from Norway. In the present investigation, we provide evidence that sarcocysts are common in tongues of Alaskan Arctic foxes suggesting that these carnivores are serving as intermediate hosts, and we also provide ultrastructure of S. arctica from the Arctic fox for the first time.

  4. Sarcocyst Development in Raccoons (Procyon lotor) Inoculated with Different Strains of Sarcocystis neurona Culture-Derived Merozoites.

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    Dryburgh, E L; Marsh, A E; Dubey, J P; Howe, D K; Reed, S M; Bolten, K E; Pei, W; Saville, W J A

    2015-08-01

    Sarcocystis neurona is considered the major etiologic agent of equine protozoal myeloencephalitis (EPM), a neurological disease in horses. Raccoon ( Procyon lotor ) is considered the most important intermediate host in the life cycle of S. neurona in the United States; S. neurona sarcocysts do mature in raccoon muscles, and raccoons also develop clinical signs simulating EPM. The focus of this study was to determine if sarcocysts would develop in raccoons experimentally inoculated with different host-derived strains of in vitro-cultivated S. neurona merozoites. Four raccoons were inoculated with strains derived from a raccoon, a sea otter, a cat, and a horse. Raccoon tissues were fed to laboratory-raised opossums ( Didelphis virginiana ), the definitive host of S. neurona . Intestinal scraping revealed sporocysts in opossums who received muscle tissue from raccoons inoculated with the raccoon-derived or the sea otter-derived isolates. These results demonstrate that sarcocysts can mature in raccoons inoculated with in vitro-derived S. neurona merozoites. In contrast, the horse and cat-derived isolates did not produce microscopically or biologically detected sarcocysts. Immunoblot analysis revealed both antigenic and antibody differences when testing the inoculated raccoons. Immunohistochemical staining indicated differences in staining between the merozoite and sarcocyst stages. The successful infections achieved in this study indicates that the life cycle can be manipulated in the laboratory without affecting subsequent stage development, thereby allowing further purification of strains and artificial maintenance of the life cycle.

  5. Sarcocystis neurona infections in sea otter (Enhydra lutris): evidence for natural infections with sarcocysts and transmission of infection to opossums (Didelphis virginiana)

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    Dubey, J.P.; Rosypal, A.C.; Rosenthal, B.M.; Thomas, N.J.; Lindsay, D.S.; Stanek, J.F.; Reed, S.M.; Saville, W.J.A.

    2001-01-01

    Although Sarcocystis neurona has been identified in an array of terrestrial vertebrates, recent recognition of its capacity to infect marine mammals was unexpected. Here, sarcocysts from 2 naturally infected sea otters (Enhydra lutris) were characterized biologically, ultrastructurally, and genetically. DNA was extracted from frozen muscle of the first of these sea otters and was characterized as S. neurona by polymerase chain reation (PCR) amplification followed by restriction fragment length polymorphism analysis and sequencing. Sarcocysts from sea otter no. 1 were up to 350 I?m long, and the villar protrusions on the sarcocyst wall were up to 1.3 I?m long and up to 0.25 I?m wide. The villar protrusions were tapered towards the villar tip. Ultrastructurally, sarcocysts were similar to S. neurona sarcocysts from the muscles of cats experimentally infected with S. neurona sporocysts. Skeletal muscles from a second sea otter failed to support PCR amplification of markers considered diagnostic for S. neurona but did induce the shedding of sporocysts when fed to a laboratory-raised opossum (Didelphis virginiana). Such sporocysts were subsequently fed to knockout mice for the interferon-gamma gene, resulting in infections with an agent identified as S. neurona on the basis of immunohistochemistry, serum antibodies, and diagnostic sequence detection. Thus, sea otters exposed to S. neurona may support the development of mature sarcocysts that are infectious to competent definitive hosts.

  6. Molecular characterization and development of Sarcocystis speeri sarcocysts in gamma interferon gene knockout mice

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    The North American opossum (Didelphis virginiana) is the definitive host for at least three named species of Sarcocystis: S. falcatula, S. neurona, and S. speeri. It appears that there may be additional undescribed species of Sarcocystis in D. virginiana feces. The South American opossums (D. albive...

  7. Sarcocystis neurona schizonts-associated encephalitis, chorioretinitis, and myositis in a two-month-old dog simulating toxoplasmosis, and presence of mature sarcocysts in muscles.

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    Dubey, J P; Black, S S; Verma, S K; Calero-Bernal, R; Morris, E; Hanson, M A; Cooley, A J

    2014-05-28

    Sarcocystis neurona is an unusual species of the genus Sarcocystis. Opossums (Didelphis virginianus, D. albiventris) are the definitive hosts and several other species, including dogs, cats, marine mammals, and horses are intermediate or aberrant hosts. Sarcocysts are not known to form in aberrant hosts. Sarcocystis neurona causes fatal disease in horses (Equine Protozoal Myeloencephalitis, EPM). There are numerous reports of fatal EPM-like infections in other species, usually with central nervous system signs and associated with the schizont stage of S. neurona. Here, we report fatal disseminated S. neurona infection in a nine-week-old golden retriever dog from Mississippi, USA. Protozoal merozoites were identified in smears of the cerebrospinal fluid. Microscopically, lesions and protozoa were identified in eyes, tongue, heart, liver, intestines, nasal turbinates, skeletal muscle and brain, which reacted intensely with S. neurona polyclonal antibodies. Mature sarcocysts were seen in sections of muscles. These sarcocysts were ultrastructurally similar to those of S. neurona from experimentally infected animals. These data suggest that the dog is another intermediate host for S. neurona. Data suggest that the dog was transplacentally infected. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Sarcocystis neurona infections in raccoons (Procyon lotor): evidence for natural infection with sarcocysts, transmission of infection to opossums (Didelphis virginiana), and experimental induction of neurologic disease in raccoons.

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    Dubey, J P; Saville, W J; Stanek, J F; Lindsay, D S; Rosenthal, B M; Oglesbee, M J; Rosypal, A C; Njoku, C J; Stich, R W; Kwok, O C; Shen, S K; Hamir, A N; Reed, S M

    2001-10-24

    Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses in the Americas and Sarcocystis neurona is the most common etiologic agent. The distribution of S. neurona infections follows the geographical distributions of its definitive hosts, opossums (Didelphis virginiana, Didelphis albiventris). Recently, cats and skunks were reported as experimental and armadillos as natural intermediate hosts of S. neurona. In the present report, raccoons (Procyon lotor) were identified as a natural intermediate host of S. neurona. Two laboratory-raised opossums were found to shed S. neurona-like sporocysts after ingesting tongues of naturally-infected raccoons. Interferon-gamma gene knockout (KO) mice fed raccoon-opossum-derived sporocysts developed neurologic signs. S. neurona was identified immunohistochemically in tissues of KO mice fed sporocysts and the parasite was isolated in cell cultures inoculated with infected KO mouse tissues. The DNA obtained from the tongue of a naturally-infected raccoon, brains of KO mice that had neurological signs, and from the organisms recovered in cell cultures inoculated with brains of neurologic KO mice, corresponded to that of S. neurona. Two raccoons fed mature S. neurona sarcocysts did not shed sporocysts in their feces, indicating raccoons are not likely to be its definitive host. Two raccoons fed sporocysts from opossum feces developed clinical illness and S. neurona-associated encephalomyelitis was found in raccoons killed 14 and 22 days after feeding sporocysts; schizonts and merozoites were seen in encephalitic lesions.

  9. Sarcocystis arctica (Apicomplexa: Sarcocystidae): ultrastructural description and its new host record, the Alaskan wolf (Canis lupus

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    Sarcocystis sarcocysts are common in muscles of herbivores but are rare in muscles of carnivores. Here, we report sarcocysts in muscle of an Alaskan wolf (Canis lupus) from Alaska, USA for the first time. Sarcocysts extracted from tongue of the wolf were up to 900 µm long, slender, and appeared to h...

  10. Sarcocystis heydorni, n. sp. (Apicomplexa: Sarcocystidae) with cattle (Bos taurus) and human (Homo sapiens) cycle.

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    Dubey, Jitender P; van Wilpe, Erna; Calero-Bernal, Rafael; Verma, Shiv Kumar; Fayer, Ronald

    2015-11-01

    Cattle (Bos taurus) are intermediate hosts for four species of Sarcocystis, namely Sarcocystis cruzi, Sarcocystis hirsuta, Sarcocystis hominis, and Sarcocystis rommeli. Of these four species, mature sarcocysts of S. cruzi are thin-walled (<1 μm), whereas S. hirsuta, S. hominis, and S. rommeli have thick walls (4 μm or more). Here, we describe a new species of Sarcocystis with thin-walled sarcocysts in cattle. Two newborn calves were fed with sporocysts from the feces of a human volunteer who had ingested raw beef. The calves were killed 111 and 222 days later. In addition to thick-walled sarcocysts of S. hominis, both calves were coinfected with a Sarcocystis species that had a thin-walled sarcocysts, distinct from S. cruzi. The sarcocysts were mature, microscopic, up to 80 μm wide, and up to 1060 μm long. By light microscopy, the sarcocyst wall was thin (<1 μm thick) and had minute protrusions. By transmission electron microscopy, the sarcocyst wall had short, conical villar protrusions (vp) that were up to 0.5 μm long and up to 0.5 μm wide, similar to type 29. The vp on the sarcocyst wall lacked microtubules but had six or more disc-shaped plaques. The ground substance layer was smooth, approximately 0.5 μm thick, and without microtubules. The bradyzoites were 8-11 μm long. The structure of the sarcocyst wall was distinct from any species of Sarcocystis reported from livestock. This unique species is named in honor of Dr. Alfred Otto Heydorn who provided the sporocysts.

  11. Sarcocystis caninum and Sarcocystis svanai n. spp. (Apicomplexa: Sarcocystidae) Associated with Severe Myositis and Hepatitis in the Domestic Dog (Canis familiaris)

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    Dubey, J. P.; Sykes, J. E.; Shelton, G. D.; Sharp, N.; Verma, S. K.; Calero-Bernal, R.; Viviano, J.; Sundar, N.; Khan, A.; Grigg, M. E.

    2014-01-01

    There are several reports of Sarcocystis sarcocysts in muscles of dogs but these species have not been named. Additionally, there are 2 reports of Sarcocystis neurona in dogs. Here, we propose 2 new names, Sarcocystis caninum, and Sarcocystis svanai for sarcocysts associated with clinical muscular sarcocystosis in 4 domestic dogs (Canis familiaris), 1 each from Montana and Colorado in the USA, and 2 from British Columbia, Canada. Only the sarcocyst stage was identified. Most of the sarcocysts identified were S. caninum. Sarcocysts were studied using light microscopy, transmission electron microscopy, and PCR. Based on collective results 2 new species, Sarcocystis caninum and Sarcocystis svanai were designated. Sarcocystis caninum and Sarcocystis svanai were structurally distinct. Sarcocystis caninum sarcocysts were up to 1.2 mm long and up to 75 μm wide. By light microscopy, the sarcocyst wall was relatively thin and smooth. By transmission electron microscopy (TEM), the sarcocyst wall “type 9”, 1–2 μm thick, and contained villar protrusions that lacked microtubules. Bradyzoites in sections were 7–9 μm long. Sarcocysts of S. svanai were few and were identified by TEM. Sarcocystis svanai sarcocysts were “type 1”, thin walled (< 0.5 μm), and the wall lacked villar protrusions but had tiny blebs that did not invaginate. DNA was extracted either from infected frozen muscle biopsies or formalin-fixed paraffin-embedded sections. Dogs were either singly infected with S. caninum or multiply co-infected with S. caninum and S. svanai (the result of a mixed infection) based on multi-locus DNA sequencing and morphology. BLASTn analysis established that the sarcocysts identified in these dogs were similar to, but not identical to S. canis or S. arctosi, parasites found to infect polar bears (Ursus maritimus) or brown bears (Ursus arctosi), respectively. However, the S. caninum sequence showed 100% identify over the 18S rRNA region sequenced to that of S. arctica

  12. Sarcocyst development in raccoons (Procyon lotor) inoculated with different strains of Sarcosytis neurona culture-derived merozoites

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    Sarcocystis neurona is considered the major etiologic agent of equine protozoal myeloencephalitis (EPM), a neurological disease in horses. Raccoon (Procyon lotor) is considered the most important intermediate host in the life cycle of S. neurona in the USA; S. neurona sarcocysts do mature in raccoon...

  13. Sarcocystis masoni, n. sp. (Apicomplexa: Sarcocystidae), and redescription of Sarcocystis aucheniae from llama (Lama glama), guanaco (Lama guanicoe) and alpaca (Vicugna pacos).

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    Moré, Gastón; Regensburger, Cristian; Gos, M Laura; Pardini, Lais; Verma, Shiv K; Ctibor, Juliana; Serrano-Martínez, Marcos Enrique; Dubey, Jitender P; Venturini, M Cecilia

    2016-04-01

    There is considerable confusion concerning the species of Sarcocystis in South American camelids (SAC). Several species names have been used; however, proper descriptions are lacking. In the present paper, we redescribe the macroscopic sarcocyst forming Sarcocystis aucheniae and describe and propose a new name, Sarcocystis masoni for the microscopic sarcocyst forming species. Muscles samples were obtained from llamas (Lama glama) and guanacos (Lama guanicoe) from Argentina and from alpacas (Vicugna pacos) and llamas from Peru. Individual sarcocysts were processed by optical and electron microscopy, and molecular studies. Microscopic sarcocysts of S. masoni were up to 800 µm long and 35-95 µm wide, the sarcocyst wall was 2·5-3·5 µm thick, and had conical to cylindrical villar protrusions (vp) with several microtubules. Each vp had 11 or more rows of knob-like projections. Seven 18S rRNA gene sequences obtained from sarcocysts revealed 95-96% identity with other Sarcocystis spp. sequences reported in the GenBank. Sarcocysts of S. aucheniae were macroscopic, up to 1·2 cm long and surrounded by a dense and laminar 50 µm thick secondary cyst wall. The sarcocyst wall was up to 10 µm thick, and had branched vp, appearing like cauliflower. Comparison of the 11 sequences obtained from individual macroscopic cysts evidenced a 98-99% of sequence homology with other S. aucheniae sequences. In conclusion, 2 morphologically and molecularly different Sarcocystis species, S. masoni (microscopic cysts) and S. aucheniae (macroscopic cysts), were identified affecting different SAC from Argentina and Peru.

  14. Molecular characterization of Sarcocystis species from Polish roe deer based on ssu rRNA and cox1 sequence analysis.

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    Kolenda, Rafał; Ugorski, Maciej; Bednarski, Michał

    2014-08-01

    Sarcocysts from four Polish roe deer were collected and examined by light microscopy, small subunit ribosomal RNA (ssu rRNA), and the subunit I of cytochrome oxidase (cox1) sequence analysis. This resulted in identification of Sarcocystis gracilis, Sarcocystis oviformis, and Sarcocystis silva. However, we were unable to detect Sarcocystis capreolicanis, the fourth Sarcocystis species found previously in Norwegian roe deer. Polish sarcocysts isolated from various tissues differed in terms of their shape and size and were larger than the respective Norwegian isolates. Analysis of ssu rRNA gene revealed the lack of differences between Sarcocystis isolates belonging to one species and a very low degree of genetic diversity between Polish and Norwegian sarcocysts, ranging from 0.1% for Sarcocystis gracilis and Sarcocystis oviformis to 0.44% for Sarcocystis silva. Contrary to the results of the ssu rRNA analysis, small intraspecies differences in cox1 sequences were found among Polish Sarcocystis gracilis and Sarcocystis silva isolates. The comparison of Polish and Norwegian cox1 sequences representing the same Sarcocystis species revealed similar degree of sequence identity, namely 99.72% for Sarcocystis gracilis, 98.76% for Sarcocystis silva, and 99.85% for Sarcocystis oviformis. Phylogenetic reconstruction and genetic population analyses showed an unexpected high degree of identity between Polish and Norwegian isolates. Moreover, cox1 gene sequences turned out to be more accurate than ssu rRNA when used to reveal phylogenetic relationships among closely related species. The results of our study revealed that the same Sarcocystis species isolated from the same hosts living in different geographic regions show a very high level of genetic similarity.

  15. Sarcocystis in moose (Alces alces): molecular identification and phylogeny of six Sarcocystis species in moose, and a morphological description of three new species.

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    Dahlgren, Stina S; Gjerde, Bjørn

    2008-06-01

    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.

  16. Molecular differentiation of Sarcocystis buffalonis and Sarcocystis levinei in water buffaloes (Bubalus bubalis) from Sarcocystis hirsuta and Sarcocystis cruzi in cattle (Bos taurus).

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    Gjerde, Bjørn; Hilali, Mosaad; Abbas, Ibrahim E

    2016-06-01

    The purpose of the present study was to obtain sarcocysts of Sarcocystis buffalonis and Sarcocystis levinei from water buffaloes and characterize the isolates by molecular methods in order to determine whether the two species were genetically different from Sarcocystis hirsuta and Sarcocystis cruzi, respectively, from cattle, which had been characterized before. About 35 macroscopically visible (3-4 × 1-2 mm) and 20 barely visible (1-3 × 0.2 mm) sarcocysts were excised from the esophagus of 18 naturally infected and freshly slaughtered adult water buffaloes at three slaughterhouses in Egypt. Genomic DNA was extracted from the sarcocysts, and all isolates were first characterized at the mitochondrial cytochrome c oxidase subunit I gene (cox1) gene through PCR amplification and direct sequencing. Selected isolates were subsequently further characterized at the 18S and 28S ribosomal (r) RNA genes and the internal transcribed spacer 1 (ITS1) region of the nuclear rDNA unit by direct sequencing or cloning. Only six of the isolated macroscopic sarcocysts belonged to S. buffalonis, whereas the others belonged to Sarcocystis fusiformis. Twelve of the smaller cysts belonged to S. levinei and seven to Sarcocystis sinensis. The characterization of the sarcocysts of S. sinensis and some of the sarcocysts of S. fusiformis have been reported before. Fifteen additional sarcocyst isolates of S. fusiformis were characterized at cox1 in the present study and found to be identical or closely similar to previous isolates. At cox1, the sequence identity between the six isolates of S. buffalonis was 99.8-100 % (two haplotypes), whereas the identity between the 12 isolates of S. levinei was 99.0-100 % (10 haplotypes). The identity between cox1 sequences of S. buffalonis and S. hirsuta (n = 56) was 92.9-93.6 % (on average 93.4 %), and the identity between cox1 sequences of S. levinei and S. cruzi (n = 22) was 92.9-94.0 % (on average 93.5 %). The phylogenetic

  17. Molecular differentiation of bovine sarcocysts.

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    Akhlaghi, Majedeh; Razavi, Mostafa; Hosseini, Arsalan

    2016-07-01

    Cattle are common intermediate hosts of Sarcocystis, and the prevalence in adult bovine muscle is close to 100 % in most regions of the world. Three Sarcocystis spp. are known to infect cattle as intermediate hosts, namely, S. cruzi, S. hirsuta, and S. hominis. The aim of the present study was the molecular identification and differentiation of these three species, Neospora caninum and Besnoitia by PCR and RFLP methods. Tissue samples were obtained from diaphragmatic muscle of 101 cattle slaughtered in Shiraz, Fars Province, Iran, for both smear preparation and DNA extraction. The samples were digested by Pepsin, washed three times with PBS solution before taking smears, fixed in absolute methanol and stained with 10 % Giemsa. The slides were examined microscopically for Sarcocystis bradyzoites and DNA was extracted from 100 mg of Sarcocystis-infected meat samples. Since the primers also bind to 18S rRNA gene of some tissue cyst-forming coccidian protozoa, DNA was also extracted from 100 μl of tachyzoite-containing suspension of N. caninum and Besnoitia isolated from goat to compare RFLP pattern. Polymerase chain reaction (PCR) was performed on DNA of samples which were microscopically positive for Sarcocystis. Five restriction enzymes Dra1, EcoRV, RsaI, AvaI, and SspI were used for RFLP and DNA of one sample from protozoa was sequenced. Based on the RFLP results, 87 (98.9 %) DNA samples were cut with DraI, indicating infection by S. cruzi. One sample (1.1 %) of PCR products of infected samples was cut only with EcoRV which showed S. hominis infection. Forty-eight samples (53.3 %) of PCR products were cut with both DraI, EcoRV, or with DraI, EcoRV, and RsaI while none of them was cut with SspI, which shows the mixed infection of both S. cruzi and S. hominis and no infection with S. hirsuta. It seems by utilizing these restriction enzymes, RLFP could be a suitable method not only for identification of Sarcocystis species but also for differentiating them

  18. Sarcocystis heydorni, n. sp. (Apicomplexa: Protozoa) with cattle (Bos taurus) and human (Homo sapiens) cycle

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    Cattle (Bos taurus) are intermediate hosts for four species of Sarcocystis, S. cruzi, S. hirsuta, S. hominis, and S. rommeli. Of these four species, mature sarcocysts of S. cruzi are thin-walled (< 1µm) whereas S. hirsuta, S. hominis, and S. rommeli have thick walls (4 µm or more). Here we describe ...

  19. Detection of zoonotic protozoa Toxoplasma gondii and Sarcocystis suihominis in wild boars from Spain

    Science.gov (United States)

    Food safety regulations require the control of presence of protozoa in meats destined for human consumption. Wild boar (Sus scrofa) meat may constitute a source of zoonoses. A 23.8% (688/2881) seroprevalence of anti-Toxoplasma gondii antibodies, and 72.2% (662/910) Sarcocystis sarcocysts prevalence ...

  20. Eosinophilic myositis resulted from Sarcocystis infection in prime marbled beef of Japanese black cattle

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    Tohru Kimura

    Full Text Available Partial changes of color (greenish to brownish were found in prime marbled beef of Japanese black cattle. The disseminated lesions of the skeletal muscles were histopathologically examined in relation to Sarcocystis infection. The lesions in the muscles showed granulomas with inflammatory cell infiltration. The sarcocysts had a distinct wall, which was radically striated by palisading villar protrusions. The sarcocyst wall was surrounded by degenerative eosinophils and necrotic muscle fibers. In conclusion, eosinophilic myositis in prime marbled beef of Japanese black cattle resulted from Sarcocystis spp. infection. The muscular lesions were characterized by the presence of granulomas and capsulated sarcocysts surrounded by numerous eosinophils. [Vet. World 2011; 4(11.000: 500-502

  1. Detection of Sarcocystis spp. infection in bobcats (Lynx rufus)

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    Verma, S. K.; Calero-Bernal, R.; Lovallo, M. J.; Sweeny, A. R.; Grigg, M. E.; Dubey, J. P.

    2015-01-01

    The protozoan Sarcocystis neurona is an important cause of severe clinical disease of horses (called equine protozoal myeloencephalitis, EPM), marine mammals, companion animals, and several species of wildlife animals in the Americas. The Virginia opossum (Didelphis virginiana) is its definitive host in the USA and other animals act as intermediate or aberrant hosts. Samples of tongue and heart from 35 bobcats hunted for fur and food from Mississippi State, USA in February, 2014 were used for the present study. Muscles were examined for Sarcocystis infection by microscopic examination of either unfixed muscle squash preparations or pepsin digests, by histopathology of fixed samples, and by molecular methods. Sarcocystis-like bradyzoites were found in digests of 14 hearts and 10 tongues of 35 bobcats. In histological sections, sarcocysts were found in 26 of 35 bobcats; all appeared relatively thin-walled similar to S. felis sarcocysts under light microscope at 1000x magnification. S. neurona-like sarcocysts having thickened villar tips were seen in unstained muscle squash of tongue of two bobcats and PCR-DNA sequencing identified them definitively as S. neurona-like parasite. DNA extracted from bradyzoites obtained from tongue and heart muscle digests was analyzed by PCR-DNA sequencing at the ITS1 locus. Results indicated the presence of S. neurona-like parasite in 26 of 35 samples. ITS1 sequences identical to S. dayspi were identified in 3 bobcats, 2 of which were also co-infected with S. neurona-like parasite. The high prevalence of sarcocysts in bobcat tissues suggested an efficient sylvatic cycle of Sarcocystis spp. in the remote regions of Mississippi State with the bobcat as a relevant intermediate host. PMID:26138150

  2. Sarcocystis fayeri in skeletal muscle of horses with neuromuscular disease.

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    Aleman, Monica; Shapiro, Karen; Sisó, Silvia; Williams, Diane C; Rejmanek, Daniel; Aguilar, Beatriz; Conrad, Patricia A

    2016-01-01

    Recent reports of Sarcocystis fayeri-induced toxicity in people consuming horse meat warrant investigation on the prevalence and molecular characterization of Sarcocystis spp. infection in horses. Sarcocysts in skeletal muscle of horses have been commonly regarded as an incidental finding. In this study, we investigated the prevalence of sarcocysts in skeletal muscle of horses with neuromuscular disease. Our findings indicated that S. fayeri infection was common in young mature horses with neuromuscular disease and could be associated with myopathic and neurogenic processes. The number of infected muscles and number of sarcocysts per muscle were significantly higher in diseased than in control horses. S. fayeri was predominantly found in low oxidative highly glycolytic myofibers. This pathogen had a high glycolytic metabolism. Common clinical signs of disease included muscle atrophy, weakness with or without apparent muscle pain, gait deficits, and dysphagia in horses with involvement of the tongue and esophagus. Horses with myositis were lethargic, apparently painful, stiff, and reluctant to move. Similar to humans, sarcocystosis and cardiomyopathy can occur in horses. This study did not establish causality but supported a possible association (8.9% of cases) with disease. The assumption of Sarcocysts spp. being an incidental finding in every case might be inaccurate.

  3. Cyst wall ultrastructure of two Sarcocystis spp. from European mouflon (Ovis ammon musimon) in Germany compared with domestic sheep.

    Science.gov (United States)

    Odening, K; Stolte, M; Walter, G; Bockhardt, I

    1995-10-01

    Muscle samples from six wild and two captive European mouflons (Ovis ammon musimon) in Germany as well as one domestic sheep from a German zoo were infected with sarcocysts (Sarcocystis: Sarcocystidae, Apicomplexa). Sarcocystis tenella and S. arieticanis were identified by light and electron microscopy. Both species are determined for the first time from wild sheep, and this is the first description of S. arieticanis from wild sheep.

  4. Sarcocystis nesbitti Infection in Human Skeletal Muscle: Possible Transmission from Snakes

    Science.gov (United States)

    Lau, Yee Ling; Chang, Phooi Yee; Tan, Chong Tin; Fong, Mun Yik; Mahmud, Rohela; Wong, Kum Thong

    2014-01-01

    Sarcocystis nesbitti is an intracellular protozoan parasite found as sarcocysts within muscle fibers of intermediate hosts (monkey and baboon). The definitive host is suspected to be the snake. We report two cases from a larger cohort of 89 patients who had fever, headache, and generalized myalgia after a trip to Pangkor Island, Malaysia. Sarcocysts were detected in skeletal muscle biopsy specimens by light and electron microscopy from these two patients. DNA sequencing based on the 18S ribosomal DNA region identified the Sarcocystis species as S. nesbitti. We also identified S. nesbitti sequences in the stools of a snake (Naja naja). Phylogenetic analysis showed that these sequences form a cluster with most of the other known Sarcocystis species for which the snake is a definitive host. We believe these two patients were likely to have symptomatic acute muscular sarcocystosis after S. nesbitti infection that may have originated from snakes. PMID:24420776

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

    Science.gov (United States)

    Arias, M; Yeargan, M; Francisco, I; Dangoudoubiyam, S; Becerra, P; Francisco, R; Sánchez-Andrade, R; Paz-Silva, A; Howe, D K

    2012-04-30

    Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs. Copyright © 2011 Elsevier B.V. All rights reserved.

  6. Morphological and molecular characteristics of four Sarcocystis spp. in Canadian moose (Alces alces), including Sarcocystis taeniata n. sp.

    Science.gov (United States)

    Gjerde, Bjørn

    2014-04-01

    Individual sarcocysts were isolated from fresh or alcohol-fixed muscle samples of two moose from Alberta, Canada, and examined by light (LM) and scanning electron microscopy (SEM) and molecular methods, comprising polymerase chain reaction (PCR) amplification and sequencing of the complete18S rRNA gene and the partial cytochrome c oxidase subunit I gene (cox1). By LM, four sarcocyst types were recognized, and the sequencing results showed that each type represented a distinct species, i.e. Sarcocystis alces, Sarcocystis alceslatrans, Sarcocystis ovalis and Sarcocystis taeniata n. sp. The finding of S. alceslatrans and S. ovalis has been reported briefly previously, but further details are provided here, including the ultrastructure of sarcoysts of S. alceslatrans as seen by SEM. The species S. alces was found for the first time in Canadian moose, whereas the finding of S. taeniata is the first record of this species in any host. The sarcocysts of S. taeniata were sac-like and about 1,000-1,100 × 60-80 μm in size. By LM, the cysts had a thin and smooth wall with no visible protrusions, whereas SEM revealed that the cyst surface had sparsely but regularly distributed, thin ribbon-like protrusions, about 2 μm long and 0.2 μm wide, lying flat against the surface and leaving most of the cyst surface naked. Similar protrusions have previously been reported from Sarcocystis grueneri in reindeer, which was found by sequence comparisons and phylogenetic analyses to be the species most closely related to S. taeniata. The phylogenetic analyses further suggested that S. taeniata, like S. alces and S. alceslatrans, use canids as definitive hosts, whereas corvid birds are known definitive hosts for S. ovalis. In contrast to the three other species found, S. taeniata displayed considerable intra-specific and intra-isolate sequence variation (substitutions, insertions/deletions) in certain regions of the 18S rRNA gene.

  7. The resurrection of a species: Sarcocystis bovifelis Heydorn et al., 1975 is distinct from the current Sarcocystis hirsuta in cattle and morphologically indistinguishable from Sarcocystis sinensis in water buffaloes.

    Science.gov (United States)

    Gjerde, Bjørn

    2016-01-01

    In the mid-1970s, it was established through transmission experiments and ultrastructural studies of sarcocysts by transmission electron microscopy (TEM) that cattle was the intermediate host of three Sarcocystis spp. using dogs, cats and humans, respectively, as definitive hosts. The cat-transmitted species with microscopic sarcocysts was initially named Sarcocystis bovifelis, but it was soon renamed Sarcocystis hirsuta, since it was considered to be identical with a previously named species. In recent years, an apparently new species has been detected in cattle in several countries by molecular methods and TEM and found by both methods to be indistinguishable from Sarcocystis sinensis in water buffaloes. This species was recently named Sarcocystis rommeli. Beginning in August 2014, a thorough review of papers comprising TEM micrographs of thick-walled sarcocysts in cattle was made in order to determine whether S. sinensis-like sarcocysts had been reported previously under other designations. Surprisingly, the review showed that the species S. bovifelis Heydorn et al., 1975 as described from cattle in Germany was S. sinensis-like and that indistinguishable sarcocysts had also been found in cattle in New Zealand and Canada in the 1980s. However, in the New Zealand study, these small sarcocysts were erroneously thought to represent developmental stages of a species with ultrastructurally similar but macroscopic sarcocysts, since the macroscopic cysts were found to be infective for cats. Thus, in the late 1980s, the cat-transmitted S. bovifelis, after having been renamed S. hirsuta, was erroneously synonymised with a second cat-transmitted species in cattle and then slid into obscurity until recently being rediscovered as a S. sinensis-like species in cattle and then named S. rommeli. Following the erroneous synonymisation, the name S. hirsuta has consistently been used for a taxon with macroscopic sarcocysts, and this usage should be continued. The name S. bovifelis

  8. Experimental infection of ponies with Sarcocystis fayeri and differentiation from Sarcocystis neurona infections in horses.

    Science.gov (United States)

    Saville, W J A; Dubey, J P; Oglesbee, M J; Sofaly, C D; Marsh, A E; Elitsur, E; Vianna, M C; Lindsay, D S; Reed, S M

    2004-12-01

    Sarcocystis neurona and Sarcocystis fayeri infections are common in horses in the Americas. Their antemortem diagnosis is important because the former causes a neurological disorder in horses, whereas the latter is considered nonpathogenic. There is a concern that equine antibodies to S. fayeri might react with S. neurona antigens in diagnostic tests. In this study, 4 ponies without demonstrable serum antibodies to S. neurona by Western immunoblot were used. Three ponies were fed 1 x 10(5) to 1 x 10(7) sporocysts of S. fayeri obtained from dogs that were fed naturally infected horse muscles. All ponies remained asymptomatic until the termination of the experiment, day 79 postinoculation (PI). All serum samples collected were negative for antibodies to S. neurona using the Western blot at the initial screening, just before inoculation with S. fayeri (day 2) and weekly until day 79 PI. Cerebrospinal fluid samples from each pony were negative for S. neurona antibodies. Using the S. neurona agglutination test, antibodies to S. neurona were not detected in 1:25 dilution of sera from any samples, except that from pony no. 4 on day 28; this pony had received 1 X 10(7) sporocysts. Using indirect immunofluorescence antibody tests (IFATs), 7 serum samples were found to be positive for S. neurona antibodies from 1:25 to 1:400 dilutions. Sarcocystis fayeri sarcocysts were found in striated muscles of all inoculated ponies, with heaviest infections in the tongue. All sarcocysts examined histologically appeared to contain only microcytes. Ultrastructurally, S. fayeri sarcocysts could be differentiated from S. neurona sarcocysts by the microtubules (mt) in villar protrusions on sarcocyst walls; in S. fayeri the mt extended from the villar tips to the pellicle of zoites, whereas in S. neurona the mt were restricted to the middle of the cyst wall. Results indicate that horses with S. fayeri infections may be misdiagnosed as being S. neurona infected using IFAT, and further research

  9. Experimental inoculation of domestic cats (Felis domesticus) with Sarcocystis neurona or S. neurona-like merozoites.

    Science.gov (United States)

    Butcher, M; Lakritz, J; Halaney, A; Branson, K; Gupta, G D; Kreeger, J; Marsh, A E

    2002-07-29

    Sarcocystis neurona is the parasite most commonly associated with equine protozoal myeloencephalitis (EPM). Recently, cats (Felis domesticus) have been demonstrated to be an experimental intermediate host in the life cycle of S. neurona. This study was performed to determine if cats experimentally inoculated with culture-derived S. neurona merozoites develop tissue sarcocysts infectious to opossums (Didelphis virginiana), the definitive host of S. neurona. Four cats were inoculated with S. neurona or S. neurona-like merozoites and all developed antibodies reacting to S. neurona merozoite antigens, but tissue sarcocysts were detected in only two cats. Muscle tissues from the experimentally inoculated cats with and without detectable sarcocysts were fed to laboratory-reared opossums. Sporocysts were detected in gastrointestinal (GI) scrapings of one opossum fed experimentally infected feline tissues. The study results suggest that cats can develop tissue cysts following inoculation with culture-derived Sarcocystis sp. merozoites in which the particular isolate was originally derived from a naturally infected cat with tissue sarcocysts. This is in contrast to cats which did not develop tissue cysts when inoculated with S. neurona merozoites originally derived from a horse with EPM. These results indicate present biological differences between the culture-derived merozoites of two Sarcocystis isolates, Sn-UCD 1 and Sn-Mucat 2.

  10. An outbreak of Sarcocystis calchasi encephalitis in multiple psittacine species within an enclosed zoological aviary.

    Science.gov (United States)

    Rimoldi, Guillermo; Speer, Brian; Wellehan, James F X; Bradway, Daniel S; Wright, Lewis; Reavill, Drury; Barr, Bradd C; Childress, April; Shivaprasad, H L; Chin, Richard P

    2013-11-01

    A total of 5 psittacine birds in an enclosed zoological exhibit, including 2 princess parrots and 3 cockatoos of 2 different species, developed severe central nervous system clinical signs over a 2-3-month period and died or were euthanized. Histologically, all birds had a lymphoplasmacytic and histiocytic encephalitis with intralesional protozoa consistent with a Sarcocystis species in addition to intramuscular tissue sarcocysts. By immunohistochemical staining, merozoites in brain and tissue cysts in muscle did not react with polyclonal antisera against Sarcocystis falcatula, Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum, or with a monoclonal antibody to S. neurona. Transmission electron microscopy on sarcocyst tissue cyst walls from 2 birds was morphologically consistent with Sarcocystis calchasi. Polymerase chain reaction (PCR) amplification and sequencing of partial 18S ribosomal RNA from muscle tissue cysts and brain schizonts from 3 birds was consistent with a clade containing S. calchasi and Sarcocystis columbae but could not distinguish these closely related Sarcocystis species. However, PCR amplification and sequencing of the internal transcribed spacer 1 RNA segment in the brain from 2 birds and muscle from 2 birds specifically identified the isolates as S. calchasi. The current report documents that multiple psittacine species are susceptible intermediate hosts of S. calchasi, and that infection can cause encephalitis resulting in significant morbidity and mortality in psittacine aviaries.

  11. Prevalence, morphology, and molecular characteristics of Sarcocystis spp. in domestic goats (Capra hircus) from Kunming, China.

    Science.gov (United States)

    Hu, Jun-Jie; Liu, Ting-Ting; Liu, Qiong; Esch, G W; Chen, Jin-Qing; Huang, Si; Wen, Tao

    2016-10-01

    Despite the importance of worldwide goat production, little is known about the prevalence of Sarcocystis spp. in domestic goats (Capra hircus) in China. The aims of the present study were to determine prevalence of Sarcocystis spp. in domestic goats in Kunming, China, as well as to identify parasite species based on morphological characteristics and DNA sequence analysis. Only microscopic sarcocysts of Sarcocystis spp. were detected in 174 of 225 goats (77.3 %). By light and transmission electron microscopy, two species, i.e., Sarcocystis capracanis and Sarcocystis hircicanis, were identified. Two sarcocysts from each of the two species were randomly selected for DNA extraction; the 18S rRNA gene (18S rRNA), the 28S rRNA gene (28S rRNA), and the mitochondrial cytochrome c oxidase subunit 1 (cox1) were amplified by the polymerase chain reaction (PCR) and subsequently sequenced. The results were compared with other previously sequenced Sarcocystis species retrieved from GenBank. There was little sequence variation between two isolates of the same species. S. capracanis was most closely related with Sarcocystis tenella; 18S rRNA, 28S rRNA, and mitochondrial cox1 sequences shared identities of 95.7-99.1, 95.3, and 92.3-93.2 % with those of S. tenella, respectively. Thus, mitochondrial cox1 sequences seem to perform better than 18S rRNA sequences or 28S rRNA sequences for identification of the two species. S. hircicanis was most closely related to Sarcocystis arieticanis, i.e., 18S rRNA and 28S rRNA sequences of the former species shared 97.2-97.4 and 95.6-96.1 % identities with those of latter, respectively. Phylogenetic analysis inferred from the three genetic markers yielded similar results and indicated the two species were within a group of Sarcocystis species with canines as known, or presumed, definitive hosts.

  12. Evidence to support horses as natural intermediate hosts for Sarcocystis neurona.

    Science.gov (United States)

    Mullaney, Thomas; Murphy, Alice J; Kiupel, Matti; Bell, Julia A; Rossano, Mary G; Mansfield, Linda S

    2005-10-10

    Opossums (Didelphis spp.) are the definitive host for the protozoan parasite Sarcocystis neurona, the causative agent of equine protozoal myeloencephalitis (EPM). Opossums shed sporocysts in feces that can be ingested by true intermediate hosts (cats, raccoons, skunks, armadillos and sea otters). Horses acquire the parasite by ingestion of feed or water contaminated by opossum feces. However, horses have been classified as aberrant intermediate hosts because the terminal asexual sarcocyst stage that is required for transmission to the definitive host has not been found in their tissues despite extensive efforts to search for them [Dubey, J.P., Lindsay, D.S., Saville, W.J., Reed, S.M., Granstrom, D.E., Speer, C.A., 2001b. A review of Sarcocystis neurona and equine protozoal myeloencephalitis (EPM). Vet. Parasitol. 95, 89-131]. In a 4-month-old filly with neurological disease consistent with EPM, we demonstrate schizonts in the brain and spinal cord and mature sarcocysts in the tongue and skeletal muscle, both with genetic and morphological characteristics of S. neurona. The histological and electron microscopic morphology of the schizonts and sarcocysts were identical to published features of S. neurona [Stanek, J.F., Dubey, J.P., Oglesbee, M.J., Reed, S.M., Lindsay, D.S., Capitini, L.A., Njoku, C.J., Vittitow, K.L., Saville, W.J., 2002. Life cycle of Sarcocystis neurona in its natural intermediate host, the raccoon, Procyon lotor. J. Parasitol. 88, 1151-1158]. DNA from schizonts and sarcocysts from this horse produced Sarcocystis specific 334bp PCR products [Tanhauser, S.M., Yowell, C.A., Cutler, T.J., Greiner, E.C., MacKay, R.J., Dame, J.B., 1999. Multiple DNA markers differentiate Sarcocystis neurona and Sarcocystis falcatula. J. Parasitol. 85, 221-228]. Restriction fragment length polymorphism (RFLP) analysis of these PCR products showed banding patterns characteristic of S. neurona. Sequencing, alignment and comparison of both schizont and sarcocyst DNA

  13. The red fox (Vulpes vulpes) and the arctic fox (Vulpes lagopus) are definitive hosts of Sarcocystis alces and Sarcocystis hjorti from moose (Alces alces).

    Science.gov (United States)

    Dahlgren, Stina S; Gjerde, Bjørn

    2010-09-01

    The aim of this study was to determine whether foxes might act as definitive hosts of Sarcocystis alces in moose. In 2 experiments, 6 silver foxes (Vulpes vulpes) and 6 blue foxes (Vulpes lagopus) were fed muscle tissue from moose containing numerous sarcocysts of S. alces, and euthanazed 7-28 days post-infection (p.i.). Intestinal mucosal scrapings and faecal samples were screened microscopically for Sarcocystis oocysts/sporocysts, which were identified to species by means of species-specific primers and sequence analysis targeting the ssu rRNA gene. All foxes in both experiments became infected with Sarcocystis; the oocysts were fully sporulated by 14 days p.i., containing sporocysts measuring 14-15 x 10 microm. Molecular identification revealed that the oocysts/sporocysts belonged to 2 species, S. alces and Sarcocystis hjorti, although sarcocysts of S. hjorti were only identified in moose subsequent to the infection of foxes. In the first experiment, all 8 foxes also became infected with a Hammondia sp. derived from moose, shedding unsporulated, subspherical oocysts, measuring 10-12 microm in diameter, from 6-7 days p.i. onwards. The study proved that canids (the red fox and arctic fox) are definitive hosts for S. alces and S. hjorti, as had been inferred from the phylogenetic position of these species.

  14. Life cycle of Sarcocystis neurona in its natural intermediate host, the raccoon, Procyon lotor.

    Science.gov (United States)

    Stanek, J F; Dubey, J P; Oglesbee, M J; Reed, S M; Lindsay, D S; Capitini, L A; Njoku, C J; Vittitow, K L; Saville, W J A

    2002-12-01

    Sarcocystis neurona causes encephalomyelitis in many species of mammals and is the most important cause of neurologic disease in the horse. Its complete life cycle is unknown, particularly its development and localization in the intermediate host. Recently, the raccoon (Procyon lotor) was recognized as a natural intermediate host of S. neurona. In the present study, migration and development of S. neurona was studied in 10 raccoons that were fed S. neurona sporocysts from experimentally infected opossums; 4 raccoons served as controls. Raccoons were examined at necropsy 1, 3, 5, 7, 10, 14, 15, 22, 37, and 77 days after feeding on sporocysts (DAFS). Tissue sections of most of the organs were studied histologically and reacted with anti-S. neurona-specific polyclonal rabbit serum in an immunohistochemical test. Parasitemia was demonstrated in peripheral blood of raccoons 3 and 5 DAFS. Individual zoites were seen in histologic sections of intestines of raccoons euthanized 1, 3, and 5 DAFS. Schizonts and merozoites were seen in many tissues 7 to 22 DAFS, particularly in the brain. Sarcocysts were seen in raccoons killed 22 DAFS. Sarcocysts at 22 DAFS were immature and seen only in skeletal muscle. Mature sarcocysts were seen in all skeletal samples, particularly in the tongue of the raccoon 77 DAFS; these sarcocysts were infective to laboratory-raised opossums. This is the first report of the complete development of S. neurona schizonts and sarcocysts in a natural intermediate host.

  15. Transmission studies with Sarcocystis idahoensis of deer mice (Peromyscus maniculatus) and gopher snakes (Pituophis melanoleucus).

    Science.gov (United States)

    Bledsoe, B

    1980-04-01

    Transmission studies with Sarcocystis idahoensis of deer mice (Peromyscus maniculatus) and gopher snakes (pituophis melanoleucus) were conducted to determine host specificity of various stages of the parasite. Sporocysts were not passed by four dogs or four cats fed infected skeletal muscle from deer mice. Seven white mice (Mus musculus) and 34 white-footed mice (Peromyscus leucopus) were negative for sarcocysts and liver meronts following oral inoculation with S. idahoensis sporocysts; however, excystation of sporocysts occurred in two white-footed mice killed four hours post inoculation (PI). A gopher snake orally inoculated with sporocysts remained negative for coccidia for two months PI. Three deer mice orally inoculated and three intraperitoneally (IP) inoculated with tachyzoites from liver meronts developed sarcocysts in their skeletal muscles similar to those seen in deer mice orally inoculated with sporocysts. Liver meronts were not found. Ten deer mice orally inoculated and 10 deer mice inoculated IP with bradyzoites from S. idahoensis sarcocysts remained negative for sarcocysts and liver meronts at necropsy 17 days PI.

  16. In the United States, negligible rates of zoonotic sarcocystosis occur in feral swine that, by contrast, frequently harbour infections with Sarcocystis miescheriana, a related parasite contracted from canids.

    Science.gov (United States)

    Calero-Bernal, R; Verma, S K; Oliveira, S; Yang, Y; Rosenthal, B M; Dubey, J P

    2015-04-01

    Transmission of pathogens between domestic and wild life animals plays an important role in epidemiology. Feral pig populations are increasing and expanding in the USA, and may constitute a risk to non-biosecure domestic pig facilities by serving as reservoirs for pathogens. We surveyed, for Sarcocystis infection, the myocardium of 1006 feral pigs (Sus scrofa) trapped or hunted in 29 states during the Comprehensive Feral Swine Disease Surveillance Program of the USDA's Animal and Plant Health Inspection Service, Wildlife Services unit during 2012-2014. Sarcocysts were detected in histological sections of 25% (251/1006) of myocardium with an average parasitic load/intensity of infection of 3.03 sarcocysts/section (1.5×0.7 cm), and higher prevalence of myocarditis in severe infections. Microscopic examination of pepsin digests of 147 hearts revealed a higher prevalence of Sarcocystis bradyzoites (49%, 72/147) than when diagnosed by histology. A fragment of Sarcocystis 18S rRNA was amplified and digested with a restriction endonuclease, revealing a pattern consistent with Sarcocystis miescheriana in all 44 selected samples. Sequencing 31 of these 44 isolates confirmed their correspondence to S. miescheriana. Thus, S. miescheriana infection, but not the zoonotic parasite Sarcocystis suihominis, appears to be prevalent and widespread in feral pigs in the USA.

  17. Short communication: Genetic variants of Sarcocystis cruzi in infected Malaysian cattle based on 18S rDNA.

    Science.gov (United States)

    Ng, Yit Han; Fong, Mun Yik; Subramaniam, Vellayan; Shahari, Shahhaziq; Lau, Yee Ling

    2015-12-01

    Sarcocystis species are pathogenic parasites that infect a wide range of animals, including cattle. A high prevalence of cattle sarcocystosis has been reported worldwide, but its status is unknown in Malaysia. This study focused on utilizing 18S rDNA to identify Sarcocystis species in Malaysian cattle and to determine their genetic variants. In this study, only Sarcocystis cruzi was detected in Malaysian cattle. The intra-species S. cruzi phylogenetic tree analysis and principal coordinate analysis (PCoA), respectively displayed two minor groups among the parasite isolates. This finding was supported by high Wright FST value (FST=0.647). The definitive hosts (dogs) may play a fundamental role in the development of S. cruzi genetic variants. Additionally, the existence of microheterogeneity within the S. cruzi merozoites and/or distinct genetic variants arisen from independent merozoites in mature sarcocysts, possibly contributed to the existence of intra-species variations within the population.

  18. Detection of Zoonotic Protozoa Toxoplasma gondii and Sarcocystis suihominis in Wild Boars from Spain.

    Science.gov (United States)

    Calero-Bernal, R; Pérez-Martín, J E; Reina, D; Serrano, F J; Frontera, E; Fuentes, I; Dubey, J P

    2016-08-01

    Food safety regulations require the control of the presence of protozoa in meats destined for human consumption. Wild boar (Sus scrofa) meat may constitute a source of zoonoses. A 23.8% (688/2881) seroprevalence of anti-Toxoplasma gondii antibodies and 72.2% (662/910) Sarcocystis sarcocysts prevalence were detected among wild boars hunted in Southwestern areas of Spain. Identity of Sarcocystis spp. was performed by RFLP-PCR and sequencing, detecting S. miescheriana (7/8) and the zoonotic S. suihominis (1/8). Risk assessment studies of these coccidian in meats destined to human consumption are needed. © 2015 Blackwell Verlag GmbH.

  19. Sarcocystis chloropusae (protozoa: Sarcocystidae) n. sp. from the common moorhen (Gallinula chloropus) from Egypt.

    Science.gov (United States)

    El-Morsey, A; El-Seify, M; Desouky, A Y; Abdel-Aziz, M M; Sakai, H; Yanai, T

    2015-07-01

    A new name Sarcocystis chloropusae is proposed for a parasite previously found in two of 25 common moorhen (Gallinula chloropus) from Brolos Lake, Egypt. Sarcocysts were microscopic, up to 650 μm long, the cyst wall was up to 4.5 μm thick, and contained villar protrusions that were up to 4 μm long and up to 2 μm wide. The villar protrusions were crowded, contained vesicles but lacked microtubules. The ground substance layer was smooth. The bradyzoites were up to 12 μm long and up to 2 μm wide. Molecular characterization and phylogenetic analysis of the (ITS-1) supported the conclusion that the Sarcocystis in G. chloropus is a distinct species.

  20. Brown-headed cowbirds (Molothrus ater) harbor Sarcocystis neurona and act as intermediate hosts.

    Science.gov (United States)

    Mansfield, L S; Mehler, S; Nelson, K; Elsheikha, H M; Murphy, A J; Knust, B; Tanhauser, S M; Gearhart, P M; Rossano, M G; Bowman, D D; Schott, H C; Patterson, J S

    2008-05-06

    We tested the hypothesis that brown-headed cowbirds (Molothrus ater) harbor Sarcocystis neurona, the agent of equine protozoal myeloencephalitis (EPM), and act as intermediate hosts for this parasite. In summer 1999, wild caught brown-headed cowbirds were collected and necropsied to determine infection rate with Sarcocystis spp. by macroscopic inspection. Seven of 381 (1.8%) birds had grossly visible sarcocysts in leg muscles with none in breast muscles. Histopathology revealed two classes of sarcocysts in leg muscles, thin-walled and thick-walled suggesting two species. Electron microscopy showed that thick-walled cysts had characteristics of S. falcatula and thin-walled cysts had characteristics of S. neurona. Thereafter, several experiments were conducted to confirm that cowbirds had viable S. neurona that could be transmitted to an intermediate host and cause disease. Specific-pathogen-free opossums fed cowbird leg muscle that was enriched for muscle either with or without visible sarcocysts all shed high numbers of sporocysts by 4 weeks after infection, while the control opossum fed cowbird breast muscle was negative. These sporocysts were apparently of two size classes, 11.4+/-0.7 microm by 7.6+/-0.4 microm (n=25) and 12.6+/-0.6 microm by 8.0+/-0 microm (n=25). When these sporocysts were excysted and introduced into equine dermal cell tissue culture, schizogony occurred, most merozoites survived and replicated long term and merozoites sampled from the cultures with long-term growth were indistinguishable from known S. neurona isolates. A cowbird Sarcocystis isolate, Michigan Cowbird 1 (MICB1), derived from thin-walled sarcocysts from cowbirds that was passaged in SPF opossums and tissue culture went on to produce neurological disease in IFNgamma knockout mice indistinguishable from that of the positive control inoculated with S. neurona. This, together with the knowledge that S. falcatula does not cause lesions in IFNgamma knockout mice, showed that cowbird

  1. Ultrastructural studies of sarcocystis cruzi (Hasselmann,1926 Wenyon, 1926 infection in cattle (Bos Taurus: Philippine cases

    Directory of Open Access Journals (Sweden)

    Claveria F.G

    2001-09-01

    Full Text Available This paper documents the first report of Sarcocysli s cruzi infection in domesticated cattle (Bos taurus in the Philippines. Fusiformshaped microscopic sarcocysts (183-578 μm long and 20-98 μm wide with distinct septae were found in the skeletal, striated and heart muscle. The sarcocyst wall or parasitophorous vacuolar membrane, 1.37-2.75 μm thick consisted of closely-packed villar protrusions 80-400 nm in dm. Middle and distal segments of VP were bent approximately 90 degrees parallel to the cyst wall surface. The villar core lacked microtubules, and at some points, the distal ends of the VP collectively formed conical tufts. Primary cyst wall had numerous 70-100 nm bubble-like undulations, and the ground substance was 0.25-0.5 μm in thickness. The ultrastructure of S. cruzi cyst wall typifies the Type 7 sarcocyst wall, and bears close similarities with the Philippine and the Vietnam strain of bubaline Sarcocystis levinei.

  2. Acute onset of encephalomyelitis with atypical lesions associated with dual infection of Sarcocystis neurona and Toxoplasma gondii in a dog.

    Science.gov (United States)

    Gerhold, Richard; Newman, Shelley J; Grunenwald, Caroline M; Crews, Amanda; Hodshon, Amy; Su, Chunlei

    2014-10-15

    A two-year-old male, neutered, basset hound-beagle mix with progressive neurological impairment was examined postmortem. Grossly, the dog had multiple raised masses on the spinal cord between nerve roots. Microscopically, the dog had protozoal myeloencephalitis. Toxoplasma gondii and Sarcocystis neurona were detected in the CNS by immunohistochemistry and polymerase chain reaction (PCR). Sarcocysts in formalin-fixed muscle were negative for Sarcocystis by PCR. Banked serum was negative for T. gondii using the modified agglutination test, suggesting an acute case of T. gondii infection or immunosuppression; however, no predisposing immunosuppressive diseases, including canine distemper, were found. To the authors' knowledge, this is the first report of dual T. gondii and S. neurona infection in a dog. Published by Elsevier B.V.

  3. Sarcocystis cruzi (Apicomplexa: Sarcocystidae no cachorro-do-mato (Cerdocyon thous Sarcocystis cruzi (Apicomplexa: Sarcocystidae in the crab-eating fox (Cerdocyon thous

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    Janaina S. Rodrigues

    2008-11-01

    Full Text Available Esporocistos de Sarcocystis foram identificados nas amostras fecais de um cachorro-do-mato. Eles foram dados por via oral para um bezerro em aleitamento, sendo observados cistos com morfologia compatível com os de Sarcocystis cruzi na musculatura cardíaca e esquelética, três meses após a infecção. Musculatura cardíaca deste bezerro foi dada para um segundo cão doméstico livre de coccídios, que eliminou esporocistos compatíveis com os de Sarcocystis em suas fezes, tendo com períodos pré-patente e patente 11 e 12 dias após a infecção respectivamente. Para comparar a morfologia dos esporocistos e cistos, um segundo cão, também livre de coccídios, foi alimentado com musculatura cardíaca de um bovino infectando naturalmente e positivo para cistos de S. cruzi. Esporocistos compatíveis com os eliminados pelo primeiro cão foram encontrados nas fezes. Apesar dos esporocistos eliminados pelo cachorro-do-mato serem significativamente diferentes dos eliminados pelos cães infectados experimentalmente, pode se considerar com base na morfologia dos esporocistos, cistos e na transmissão biológica que a espécie encontrada nas fezes do cachorro-do-mato é Sarcocystis cruzi.Sporocysts of Sarcocystis were identified in feces samples of a crab-eating fox, and were orally given to a suckling calf; after 3 months of infection, sarcocysts morphologically similar to Sarcocystis cruzi were observed in cardiac and skeletal striated muscles. The cardiac muscles of this calf were orally given to a puppy free of coccidia, that shed sporocysts in its feces.with a prepatent and patent period of 11 and 12 days after infection, respectively. To compare the morphology of the sporocysts and cysts, a second puppy was fed on bovine cardiac muscles infected naturally, and sporocysts identical to those shed by the first dog were recovered from its feces. In spite of the significant difference between sporocysts found in the mucosa of the crab-eating fox and

  4. A Survey on Sarcocystis Infection Rate in Slaughtered Cattle and Sheep by Macroscopic Inspection and Pepsin Digestion Methods in Hamadan Abattoir, Iran, 2014

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    F. Parandin

    2015-10-01

    Full Text Available Introduction & Objective: 130 heteroxenous species of sarcosytis with different life cycle and pathogenesis have been recognized. The pathogenic species for humans are S. hominis from cattle and S. suihominis from pig that humans are definitive and cattle and pig are intermedi-ate hosts. Some species of Sarcocystis can cause important economic loss and disease in livestock, and health issues in humans. The aim of this study was to determine the prevalence of Sarcocystis infection in slaughtered Cattle and sheep in Hamadan, west of Iran. Materials & Methods: In this cross sectional descriptive study a total of 324 cattle and 334 sheep carcasses were examined using naked eye inspection for macroscopic Sarcocysts, and digestion method, for microscopic types of parasite. Muscles from thigh, heart, tongue, esophagus, diaphragm and costal muscles were examined. All carcasses examined by naked eyes and tissues were minced and poured in digestion medium separately and sediment was examined microscopically. Results: The prevalence of microscopic Sarcocystis in cattle was detected in 100% and there was no macroscopic cyst in examined carcasses. However, the prevalence of microscopic Sarcocystis in the sheep was also 100% and the sarcocysts were found in the 48.34 % of esophagus and 29.49% of diaphragm muscles by naked eyes inspection. Conclusion: The digestion is found the most sensitive method for diagnosis of Sarcocystis. Al-though 100% of muscles were found infected but the majority of the cysts in the sheep and all in the cattle were as microcysts. That means, the meat should be cooked sufficiently irrespec-tive of meat inspection results. (Sci J Hamadan Univ Med Sci 2015; 22 (3: 210-216

  5. Morphologic identification of a new Sarcocystis sp. in the common moorhen (Gallinula chloropus ) (Aves: Gruiformes: Rallidae) from Brolos Lake, Egypt.

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    El-Morsey, Ahmed; El-Seify, Mahmoud; Desouky, Abdel-Razik Y; Abdel-Aziz, Mohamed M; Sakai, Hiroki; Yanai, Tokuma

    2014-01-01

    Sarcocystis species are among the most common and widespread protozoan parasites of mammals and birds. The current study provides the first record of infection with Sarcocystis species in the common moorhens from Brolos Lake, KafrElsheikh province, Egypt. Morphology of the parasite cysts was described using light and transmission electron microscopy. Out of 25 examined birds, sarcocysts were found in neck, thigh, and legs muscles of two birds. The cysts were microscopic and measured 150-650 μm in length×45-185 μm in width. Histologically, the sarcocyst wall appeared striated and characterized by the presence of radial spines. Ultrastructurally, it measured 2-4.5 μm in thickness and had irregularly shaped crowded finger-like villar protrusions that measured 1.5-4 μm in length and up to 0.4-2 μm in width with the presence of dense electron ground substance of 200-750 nm thick. Several septa derived from the ground substance were present and divided the cyst into compartments containing both bradyzoites and metrocytes. The bradyzoites were banana-shaped and measured 6-12 × 1-2 μm with centrally or posteriorly located nuclei. The ultrastructural features of the cyst wall belonged to type 10 cyst wall according the classification of Dubey et al. (1989) and Dubey and Odening (2001).

  6. Accipiter hawks (Accipitridae) confirmed as definitive hosts of Sarcocystis turdusi, Sarcocystis cornixi and Sarcocystis sp. ex Phalacrocorax carbo.

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    Mayr, Sylvia L; Maier, Kristina; Müller, Jana; Enderlein, Dirk; Gruber, Achim D; Lierz, Michael

    2016-08-01

    Sarcocystis is a large genus of protozoan parasites with complex heteroxenous life cycles. For many species, either the intermediate or the definitive host is still unknown. In this study, 116 Accipiter hawks (Eurasian sparrowhawks and northern goshawks) were investigated for the presence of Sarcocystis spp. in their intestinal tract or their faeces. To gain a wide distribution, samples were collected throughout Germany within 2 years. It was possible to detect Sarcocystis-like oocysts in 65 samples. Sequencing of the ITS region or species-specific PCR identified 33 samples as Sarcocystis turdusi/Sarcocystis sp. ex A. nisus (18), Sarcocystis calchasi (6), Sarcocystis columbae (3), Sarcocystis cornixi (3) and Sarcocystis sp. ex Phalacrocorax carbo (3). Besides the known infestation with S. columbae, S. sp. ex A. nisus and S. calchasi the Accipiter hawks were thereby confirmed as definitive host of S. turdusi, S. cornixi and S. sp. ex Phalacrocorax carbo for the first time.

  7. Sarcocystis neurona manipulation using culture-derived merozoites for bradyzoite and sporocyst production.

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    Chaney, Sarah B; Marsh, Antoinette E; Lewis, Stephanie; Carman, Michelle; Howe, Daniel K; Saville, William J; Reed, Stephen M

    2017-04-30

    Equine protozoal myeloencephalitis (EPM) remains a significant central nervous system disease of horses in the American continents. Sarcocystis neurona is considered the primary causative agent and its intermediate life stages are carried by a wide host-range including raccoons (Procyon lotor) in North America. S. neurona sarcocysts mature in raccoon skeletal muscle and can produce central nervous system disease in raccoons, mirroring the clinical presentation in horses. The study aimed to develop laboratory tools whereby the life cycle and various life stages of S. neurona could be better studied and manipulated using in vitro and in vivo systems and compare the biology of two independent isolates. This study utilized culture-derived parasites from S. neurona strains derived from a raccoon or from a horse to initiate raccoon infections. Raccoon tissues, including fresh and cryopreserved tissues, were used to establish opossum (Didelphis virginiana) infections, which then shed sporocyts with retained biological activity to cause encephalitis in mice. These results demonstrate that sarcocysts can be generated using in vitro-derived S. neurona merozoites, including an isolate originally derived from a naturally infected horse with clinical EPM. This study indicates the life cycle can be significantly manipulated in the laboratory without affecting subsequent stage development, allowing further purification of strains and artificial maintenance of the life cycle. Published by Elsevier B.V.

  8. Parasite distribution and early-stage encephalitis in Sarcocystis calchasi infections in domestic pigeons (Columba livia f. domestica).

    Science.gov (United States)

    Maier, Kristina; Olias, Philipp; Enderlein, Dirk; Klopfleisch, Robert; Mayr, Sylvia L; Gruber, Achim D; Lierz, Michael

    2015-01-01

    Pigeon protozoal encephalitis is a biphasic, neurologic disease of domestic pigeons (Columba livia f. domestica) caused by the apicomplexan parasite Sarcocystis calchasi. Despite severe inflammatory lesions of the brain, associated parasitic stages have only rarely been identified and the cause of the lesions is still unclear. The aim of this study was therefore to characterize the tissue distribution of S. calchasi within pigeons between the two clinical phases and during the occurrence of neurological signs. For this purpose, a semi-quantitative real-time polymerase chain reaction (PCR) was developed. Forty-five domestic pigeons were infected orally (via a cannula into the crop) with 200 S. calchasi sporocysts and euthanized in groups of three pigeons at intervals of 2 to 10 days over a period of 61 days. Tissue samples including brain and skeletal muscle were examined by histology, immunohistochemistry, and PCR. Schizonts were detected in the liver of one pigeon at day 10 post infection. A mild encephalitis was detected at day 20 post infection, around 4 weeks before the onset of neurological signs. At the same time, immature sarcocysts were present in the skeletal muscle. In seven pigeons a few sarcocysts were identified in the brain, but not associated with any lesion. These results suggest that the encephalitis is induced at a very early stage of the S. calchasi lifecycle rather than in the chronic phase of pigeon protozoal encephalitis. Despite the increasing severity of lesions in the central nervous system, the amount of sarcocysts did not increase. This supports the hypothesis of a delayed-type hypersensitivity response as the cause of the encephalitis. The study also demonstrated that S. calchasi DNA is detectable in tissues negative by histological methods, indicating a higher sensitivity of the real-time PCR.

  9. Occurrence and first molecular characterization of Sarcocystis spp. in wild boars (Sus scrofa) and domestic pigs (Sus scrofa domesticus) in Romania: Public health significance of the isolates.

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    Imre, Kálmán; Sala, Claudia; Morar, Adriana; Imre, Mirela; Ciontu, Cătălin; Chisăliță, Ion; Dudu, Andreea; Matei, Marius; Dărăbuș, Gheorghe

    2017-03-01

    Domestic and wild pigs, as intermediate hosts, can harbor tissue cysts of three Sarcocystis species namely S. miescheriana, S. suihominis and S. porcifelis. Out of them, S. suihominis is zoonotic. Romania is a country with high consumption of raw and/or undercooked traditional pork products. This fact may greatly favor the acquiring of the zoonotic Sarcocystis infections by humans, as definitive host. Based on this consideration and in order to investigate the occurrence and public health significance of Sarcocystis spp. in two western counties (Caraş-Severin and Timiş) of Romania, a total of 165 heart samples from hunted wild boars (Sus scrofa, n=101) and home slaughtered domestic pigs (Sus scrofa domesticus, n=64) were screened using microscopic fresh examination and molecular methods. Microscopic examination revealed the presence of sarcocysts in 60.4% of wild boars, and 23.4% of domestic pigs. Genetic characterization of isolates through the PCR-RFLP procedure, targeting the 18S rRNA gene, was successfully achieved for all microscopically positive samples, indicating the presence of a single species, S. miescheriana, in both hosts. The identity of 13 selected S. miescheriana isolates was also confirmed through sequencing. The tested hosts older than 27 months were found to be significantly higher infected (p<0.05) with Sarcocystis than the 6 to ≤27months age group. Although the human infective S. suihominis has not been registered, for a more reliable epidemiological picture, further molecular studies enrolling a larger number of animals and diagnosis on human intestinal Sarcocystis infections are still necessary.

  10. Ultrastructure of the cysts of Sarcocystis rangi from skeletal muscle of reindeer (Rangifer tarandus tarandus

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    Bjørn Gjerde

    1985-05-01

    Full Text Available Mature muscle cysts of Sarcocystis rangi from Rangifer tarandus were examined by transmission electron microscopy. The long and slender cysts were located within skeletal muscle cells, and were bounded by a unit membrane, the cyst membrane. The cysts were provided with closely spaced flexible, hairlike surface processes, measuring up to 12.6 |im in length and 0.3 to 0.6 \\lm in diameter. The projections had a smooth surface, whereas the cyst membrane formed numerous hexagonally packed vesicular invaginations between the bases of the projections. The cyst membrane was reinforced by an underlying thin layer of electron-dense material, except at the points where it was invaginated. Cyst ground substance formed a thin layer at the periphery of the cysts, filled the core of the projections, and formed thin septa that divided the interior of the cysts into numerous compartments. Most compartments contained a large number of tightly packed cystozoites, whereas a few metrocytes were forund in each of a few compartments at the periphery of the cysts. Some of the cystozoites multiplied by endodyogeny. The metrocytes displayed a vacuolation of their cytoplasm. The cysts of S. rangi were similar in surface morphology to the sarcocysts of certain other Sarcocystis species reported from other intermediate hosts.Ultrastrukturen til cyster av Sarcocystis rangi frå skjelettmuskulaturen hos rein.Abstract in Norwegian / Samandrag: Muskelcyster av S. rangi frå rein vart undersøkt ved transmisjonselektronmikroskopi. Dei lange cystene låg intracellulært i skjelettmuskelceller, og var avgrensa av ein elementærmembran, cystemembranen. Cystene var utstyrt med talrike hårliknande overflateprosessar, som strekte seg langsetter cysteoverflata. Prcsessane var opptil 12.6 Hm lange, og målte 0.3 til 0.6 \\lm i diameter. Prosessane hadde ei glatt overflate, medan cystemembranen danna talrike regelmessige ordna, små invaginasjonar innimellom basis av prosessane

  11. Molecular characterisation of three regions of the nuclear ribosomal DNA unit and the mitochondrial cox1 gene of Sarcocystis fusiformis from water buffaloes (Bubalus bubalis) in Egypt.

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    Gjerde, Bjørn; Hilali, Mosaad; Mawgood, Sahar Abdel

    2015-09-01

    A total of 33 macroscopically visible (3-11 × 1-5 mm) sarcocysts of Sarcocystis fusiformis were excised from the oesophagus of 12 freshly slaughtered water buffalos in Giza, Egypt. Genomic DNA was extracted from the sarcocysts, and all isolates were characterised at the mitochondrial cytochrome c oxidase subunit I (cox1) gene through PCR amplification and direct sequencing, whereas a few selected isolates were characterised at the 18S and 28S ribosomal (r) RNA genes and the internal transcribed spacer 1 (ITS1) region of the nuclear rDNA unit following cloning. Among the 33 cox1 sequences (1,038-bp long), there was a total of 13 haplotypes, differing from each other by one to seven substitutions and sharing an identity of 99.3-99.9 %. In comparison, the sequence identity was 98.8-99.0 % among eight complete 18S rRNA gene sequences (1,873-1,879-bp long), 98.1-100 % among 28 complete ITS1 sequences (853-864-bp long) and 97.4-99.6 % among five partial 28S rRNA gene sequences (1,607-1,622 bp). At the three nuclear loci, the intraspecific (and intra-isolate) sequence variation was due to both substitutions and indels, which necessitated cloning of the PCR products before sequencing. Some additional clones of the 18S and 28S rRNA genes were highly divergent from the more typical clones, but the true nature of these aberrant clones could not be determined. Sequence comparisons and phylogenetic analyses based on either 18S rRNA gene or cox1 nucleotide sequences, placed S. fusiformis closest to Sarcocystis cafferi from the African buffalo, but only the analyses based on cox1 data separated the two taxa clearly from each other and showed that they were separate species (monophyletic clusters and 93 % sequence identity at cox1 versus interleaved sequences and 98.7-99.1 % sequence identity at the 18S rRNA gene). Two cats experimentally infected with sarcocysts of S. fusiformis started shedding small numbers of sporocysts 8-10 days post-infection (dpi) and were euthanized 15

  12. Sarcocystis neurona encephalitis in a dog.

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    Cooley, A J; Barr, B; Rejmanek, D

    2007-11-01

    A 1.5-year-old male Feist dog was presented to a veterinarian for reluctance to stand on the hind legs. Treatment included dexamethasone and resulted in a favorable initial response, but posterior paresis returned and progressed to recumbency, hyperesthesia, and attempts to bite the owner. The dog was euthanized. The brain was negative for rabies by fluorescent antibody analysis. Multiple foci of encephalitis were found in the cerebrum and particularly in the cerebellum. Protozoa morphologically consistent with Sarcocystis sp. were identified at sites of intense inflammation and malacia. Additionally, multiple schizonts were identified in areas without inflammation. Immunohistochemistry using both polyclonal and monoclonal antibodies specific for Sarcocystis neurona was strongly positive. No reaction to polyclonal antisera for Toxoplasma gondii or Neospora caninum was found. Polymerase chain reaction confirmed that the protozoa were S. neurona. Additional aberrant hosts for S. neurona other than horses have been identified, but S. neurona encephalitis has not been documented previously in the dog.

  13. Genetic assemblage of Sarcocystis spp. in Malaysian snakes

    Science.gov (United States)

    2013-01-01

    Background Sarcocystis species are protozoan parasites with a wide host range including snakes. Although there were several reports of Sarcocytis species in snakes, their distribution and prevalence are still not fully explored. Methods In this study, fecal specimens of several snake species in Malaysia were examined for the presence of Sarcocystis by PCR of 18S rDNA sequence. Microscopy examination of the fecal specimens for sporocysts was not carried as it was difficult to determine the species of the infecting Sarcocystis. Results Of the 28 snake fecal specimens, 7 were positive by PCR. BLASTn and phylogenetic analyses of the amplified 18S rDNA sequences revealed the snakes were infected with either S. nesbitti, S. singaporensis, S. zuoi or undefined Sarcocystis species. Conclusion This study is the first to report Sarcocystis infection in a cobra, and S. nesbitti in a reticulated python. PMID:24010903

  14. Molecular evidence of Sarcocystis species in captive snakes in Japan.

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    Abe, Niichiro; Matsubara, Katsuki; Tamukai, Kenichi; Miwa, Yasutsugu; Takami, Kazutoshi

    2015-08-01

    Sarcocystis nesbitti, using snakes as the definitive host, is a causative agent of acute human muscular sarcocystosis in Malaysia. Therefore, it is important to explore the distribution and prevalence of S. nesbitti in snakes. Nevertheless, epizootiological information of S. nesbitti in snakes remains insufficient because few surveys have assessed Sarcocystis infection in snakes in endemic countries. In Japan, snakes are popular exotic pet animals that are imported from overseas, but the degree of Sarcocystis infection in them remains unclear. The possibility exists that muscular sarcocystosis by S. nesbitti occurs in contact with captive snakes in non-endemic countries. For a total of 125 snake faecal samples from 67 snake species collected at animal hospitals, pet shops and a zoo, this study investigated the presence of Sarcocystis using polymerase chain reaction (PCR) for the 18S ribosomal RNA gene (18S rDNA). Four (3.2%) faecal samples were positive by PCR. Phylogenetic analysis of the 18S rDNA sequences obtained from four amplification products revealed one isolate from a beauty snake (Elaphe taeniura), Sarcocystis zuoi, which uses rat snakes as the definitive host. The isolate from a Macklot's python (Liasis mackloti) was closely related with unidentified Sarcocystis sp. from reticulated pythons in Malaysia. The remaining two isolates from tree boas (Corallus spp.) were closely related with Sarcocystis lacertae, Sarcocystis gallotiae and unidentified Sarcocystis sp. from smooth snakes, Tenerife lizards and European shrews, respectively. This report is the first of a study examining the distribution of Sarcocystis species in captive snakes in Japan.

  15. Modified use of methylene blue in the tissue compression technique to detect sarcocysts in meat-producing animals.

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    Ng, Yit Han; Subramaniam, Vellayan; Lau, Yee Ling

    2015-11-30

    Sarcocystosis in meat-producing animals is a major cause of reduced productivity in many countries, especially those that rely on agriculture. Although several diagnostic methods are available to detect sarcocystosis, many are too time-consuming for routine use in abattoirs and meat inspection centers, where large numbers of samples need to be tested. This study aimed to compare the sensitivity of the methylene blue tissue preparation, unstained tissue preparation and nested PCR in the detection of sarcocysts in tissue samples. Approximately three-fold more sarcocysts were detected in methylene blue-stained tissue compared to unstained controls (McNemar's test: Pmethylene blue can be used in tissue compression as a rapid, safe, and inexpensive technique for the detection of ruminant sarcocystosis in abattoirs. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. A review of Sarcocystis spp. shed by opossums (Didelphis spp. in Brazil

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    Samantha Yuri Oshiro Branco Valadas

    2016-08-01

    Full Text Available South American opossums are the definitive hosts of Sarcocystis neurona, Sarcocystis falcatula, Sarcocystis speeri and Sarcocystis lindsayi. The sporocysts of these species of Sarcocystis are morphologically similar and methods like infectivity and pathogenicity for intermediate hosts (immunodeficient mice and psittacine birds and molecular tools are used for identification. Opossums are synanthropic wild animals, and widely distributed in Brazilian territory. Previous studies have shown high environmental contamination with S. neurona sporocysts in several Brazilian regions. This paper reviews information on Sarcocystis spp. shed by various opossum species and its occurrence in Brazil.

  17. Clinical Sarcocystis neurona, Sarcocystis canis, Toxoplasma gondii, and Neospora caninum infections in dogs.

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    Dubey, J P; Chapman, Jennifer L; Rosenthal, Benjamin M; Mense, M; Schueler, Ronald L

    2006-04-15

    Sarcocystis neurona, Sarcocystis canis, Toxoplasma gondii, and Neospora caninum are related apicomplexans that can cause systemic illness in many species of animals, including dogs. We investigated one breeder's 25 Basset Hounds for these infections. In addition, tissues from dogs and other non-canine hosts previously reported as S. canis infections were studied retrospectively. Schizonts resembling those of S. neurona, and recognized by polyclonal rabbit anti-S. neurona antibodies, were found in six of eight retrospective cases, as well as in two additional dogs (one Basset Hound, one Springer Spaniel) not previously reported. S. neurona schizonts were found in several tissues including the central nervous system, lungs, and kidneys. Fatal toxoplasmosis was diagnosed in an adult dog, and neosporosis was diagnosed in an adult and a pup related to the one diagnosed with S. neurona. No serological reactivity to S. neurona antibodies occurred when S. canis-like liver schizonts were retrospectively assayed from two dogs, a dolphin, a sea lion, a horse, a chinchilla, a black or either of two polar bears. Sequencing conserved (18S) and variable (ITS-1) portions of nuclear ribosomal DNA isolated from the schizont-laden liver of a polar bear distinguished it from all previously characterized species of Sarcocystis. We take this genetic signature as provisionally representative of S. canis, an assumption that should be tested with future sequencing of similar liver infections in other mammalian hosts. These findings further extend the uncharacteristically broad intermediate host range for S. neurona, which also causes a neurologic disease in cats, mink, raccoons, skunks, Pacific harbor seals, ponies, zebras, lynxes, and sea otters. Further work is necessary to delineate the causative agent(s) of other cases of canine sarcocystosis, and in particular to specify the attributes of S. canis, which corresponds morphologically to infections reported from wide range of terrestrial

  18. Intra-uterine exposure of horses to Sarcocystis spp. antigens

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    A.M. Antonello

    2016-04-01

    Full Text Available The aim of this study was to examine the intra-uterine exposure to Sarcocystis spp. antigens, determining the number of foals with detectable concentrations of antibodies against these agents in the serum, before colostrum ingestion and collect data about exposure of horses to the parasite. Serum samples were collected from 195 thoroughbred mares and their newborns in two farms from southern Brazil. Parasite specific antibody responses to Sarcocystis antigens were detected using the indirect immunofluorescent antibody test (IFAT and immunoblot analysis. In 84.1% (159/189 of the pregnant mares and in 7.4% (14/189 of foals we detected antibodies anti-Sarcocystis spp. by IFAT. All samples seropositive from foals were also positive in their respective mares. Serum samples of seropositive foals by IFAT, showed no reactivity on the immunoblot, having as antigens S. neurona merozoites. In conclusion, the intra-uterine exposure to Sarcocystis spp. antigens in horses was demonstrated, with occurrence not only in mares, but also in their foals, before colostrum ingestion these occurrences were reduced.

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

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    Zoll, W M; Needle, D B; French, S J; Lim, A; Bolin, S; Langohr, I; Agnew, D

    2015-01-01

    Two neonatal male red panda (Ailurus fulgens) littermates were submitted for necropsy examination. One animal was found dead with no prior signs of illness; the other had a brief history of laboured breathing. Post-mortem examination revealed disseminated protozoal infection. To further characterize the causative agent, transmission electron microscopy (TEM), immunohistochemistry (IHC), polymerase chain reaction (PCR) and amplification and nucleic acid sequencing were performed. IHC was negative for Toxoplasma gondii and Neospora caninum, but was positive for a Sarcocystis spp. TEM of cardiac muscle and lung revealed numerous intracellular apicomplexan protozoa within parasitophorous vacuoles. PCR and nucleic acid sequencing of partial 18S rRNA and the internal transcribed spacer (ITS)-1 region confirmed a Sarcocystis spp. that shared 99% sequence homology to Sarcocystis neurona and Sarcocystis dasypi. This represents the first report of sarcocystosis in red pandas. The histopathological, immunohistochemical, molecular and ultrastructural findings are supportive of vertical transmission resulting in fatal disseminated disease. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Survey on Sarcocystis in bovine carcasses slaughtered at the municipal abattoir of El-Kharga, Egypt

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    Ali Meawad Ahmed

    2016-12-01

    Full Text Available Aim: The main objectives of this study were to determine the incidence of Sarcocystis sp. infection in cattle and buffalo carcasses slaughtered at El-Kharga abattoir, New Valley Governorate, Egypt. Materials and Methods: The slaughtered animals were daily inspected for Sarcocystis macrocysts through a year (2015. Macroscopic Sarcocystis was detected from a total of 2120 cattle and buffalo carcasses. In addition, 100 meat samples were collected from female cattle and buffalo (50 each and were examined microscopically for sarcocystosis. Results: The overall incidence of Sarcocystis macrocyst among bovine carcasses was 159/2120 (7.5%. Total incidence in cattle was 149/2000 (7.45%, whereas it was 10/120 (8.33% in buffalo carcasses. Concerning gender, the overall prevalence of Sarcocystis infection was 127/1790 (7.09% in male and 32/330 (9.69% in females bovine carcasses. The highest detection rate of Sarcocystis lesions was from the esophagus (76.3% followed by throat muscles (35.3%, tongue (33.8%, and diaphragm muscles (18.71%. Macrocysts from cattle were identified to Sarcocystis hirsuta, whereas Sarcocystis fusiformis was identified from buffalo carcasses. By microscopic examination, 18 (36% of 50 female cattle carcasses harbor Sarcocystis sp., whereas 11 (22% of buffalo carcasses were harbored Sarcocystis microcysts. Conclusion: A high incidence of Sarcocystis infection was detected among slaughtered bovines in El-Kharga abattoir, Egypt. Sarcocystis macrocysts were a higher incidence in female elder animals macrocysts were identified to S. hirsuta in cattle and S. fusiformis in buffaloes. Sarcocystosis constitute a major cause of economic losses at El-Kharga abattoir. Beef meat may carry health risks to consumers.

  1. Sarcocystis neurona and Neospora caninum in Brazilian opossums (Didelphis spp.): Molecular investigation and in vitro isolation of Sarcocystis spp.

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    Gondim, Leane S Q; Jesus, Rogério F; Ribeiro-Andrade, Müller; Silva, Jean C R; Siqueira, Daniel B; Marvulo, Maria F V; Aléssio, Felipe M; Mauffrey, Jean-François; Julião, Fred S; Savani, Elisa San Martin Mouriz; Soares, Rodrigo M; Gondim, Luís F P

    2017-08-30

    Sarcocystis neurona and Neospora spp. are protozoan parasites that induce neurological diseases in horses and other animal species. Opossums (Didelphis albiventris and Didelphis virginiana) are definitive hosts of S. neurona, which is the major cause of equine protozoal myeloencephalitis (EPM). Neospora caninum causes abortion in cattle and infects a wide range of animal species, while N. hughesi is known to induce neurologic disease in equids. The aims of this study were to investigate S. neurona and N. caninum in tissues from opossums in the northeastern Brazil, and to isolate Brazilian strains of Sarcocystis spp. from wild opossums for comparison with previously isolated strains. Carcasses of 39 opossums from Bahia state were available for molecular identification of Sarcocystis spp. and N. caninum in their tissues, and for sporocyst detection by intestinal scraping. In addition, Sarcocystis-like sporocysts from nine additional opossums, obtained in São Paulo state, were tested. Sarcocystis DNA was found in 16 (41%) of the 39 opossums' carcasses; N. caninum DNA was detected in tissues from three opossums. The sporocysts from the nine additional opossums from São Paulo state were tested by bioassay and induced infection in nine budgerigars, but in none of the gamma-interferon knockout mice. In vitro isolation was successful using tissues from all nine budgerigars. The isolated strains were maintained in CV-1 and Vero cells. Three of nine isolates presented contamination in cell culture and were discarded. Analysis of six isolates based on five loci showed that these parasites were genetically different from each other and also distinct from S. neurona, S. falcatula, S. lindsayi, and S. speeri. In conclusion, opossums in the studied regions were infected with N. caninum and Sarcocystis spp. and represent a potential source of infection to other animals. This is the first report of N. caninum infection in tissues from black-eared opossum (D. aurita or D

  2. Systems based analysis of the Sarcocystis neurona genome identifies pathways that contribute to a heteroxenous life cycle

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    Sarcocystis neurona is a member of the Coccidia, a clade of single-celled parasites of medical and veterinary importance including Eimeria, Sarcocystis, Neospora and Toxoplasma. Unlike Eimeria, a single host enteric pathogen, Sarcocystis, Neospora and Toxoplasma are two host parasites that infect an...

  3. Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and Sarcocystis canis-like infections in marine mammals.

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    Dubey, J P; Zarnke, R; Thomas, N J; Wong, S K; Van Bonn, W; Briggs, M; Davis, J W; Ewing, R; Mense, M; Kwok, O C H; Romand, S; Thulliez, P

    2003-10-30

    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).

  4. Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and Sarcocystis canis-like infections in marine mammals

    Science.gov (United States)

    Dubey, J.P.; Zarnke, R.; Thomas, N.J.; Wong, S.K.; Vanbonn, W.; Briggs, M.; Davis, J.W.; Ewing, R.; Mense, M.; Kwok, O.C.H.; Romand, S.; Thulliez, P.

    2003-01-01

    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 a?Y1:25) 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 (Zalophus californianus), 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).

  5. Serologic responses of cats against experimental Sarcocystis neurona infections.

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    Dubey, J P; Lindsay, D S; Saville, W J A

    2002-08-02

    Sarcocystis neurona is the most important cause of a neurologic disease of horses, equine protozoal myeloencephalitis (EPM). Cats and other carnivores can act as its intermediate hosts and horses are aberrant hosts. Little is known of the sero-epidemiology of S. neurona infections in cats. In the present study, antibodies to S. neurona were evaluated by the S. neurona agglutination test (SAT). Cats fed sporocysts from the feces of naturally infected opossums or inoculated intramuscularly with S. neurona merozoites developed high levels (> or =1:4000) of SAT antibodies. Antibodies to S. neurona were not found in a cat inoculated with merozoites of the closely related parasite, Sarcocystis falcatula. These results should be useful in studying sero-epidemiology of S. neurona infections in cats.

  6. High prevalence of Sarcocystis calchasi sporocysts in European Accipiter hawks.

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    Olias, Philipp; Olias, Lena; Krücken, Jürgen; Lierz, Michael; Gruber, Achim D

    2011-02-10

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

  7. Investigação de anticorpos contra Sarcocystis neurona e Sarcocystis cruzi em equinos

    Directory of Open Access Journals (Sweden)

    A. M. Antonello

    2015-10-01

    Full Text Available ABSTRACTSarcocystis neurona is the primary agent for Equine Protozoal Myeloencephalitis (EPM, important neurological disease characterized by behavior or muscular changes, that impairs animal performance and husbandry. Sarcocystis cruzi is a pathogen related to myositis in cattle. Although related the life cycles of the parasites are distinct. S. neurona has opossums (Didelphis spp. and S. cruzi, dogs as definitive hosts. However, S. neurona and S. cruzi may undergo cross-reactivity in serological tests, interfering on results of EPM ante-mortem diagnostic tests. In the present study, serology of 189 mares was performed by indirect immunofluorescence antibody test, using antigens of S. neurona and S. cruzi in order to assess the exposure degree of animals to antigens. Analyzing the results, it was observed that most of the animals (84.13% reacted with at least one protozoal species and the number of animals which showed antibodies against S. cruzi was greater than S. neurona (80.42% and 33.86%, respectively and a third of seropositive animals reacted to antigens of both species.

  8. Modest genetic differentiation among North American populations of Sarcocystic neurona may reflect expansion in its geographic range

    Science.gov (United States)

    Sundar, N.; Asmundsson, I.M.; Thomas, N.J.; Samuel, M.D.; Dubey, J.P.; Rosenthal, B.M.

    2008-01-01

    Sarcocystis neurona is an important cause of neurological disease in horses (equine protozoal myeloencephalitis, EPM) and sea otters in the United States. In addition, EPM-like disease has been diagnosed in several other land and marine mammals. Opossums are its only definitive hosts. Little genetic diversity among isolates of S. neurona from different hosts has been reported. Here, we used 11 microsatellites to characterize S. neurona DNA isolated from natural infections in 22 sea otters (Enhydra lutris) from California and Washington and in 11 raccoons (Procyon lotor) and 1 striped skunk (Mephitis mephitis) from Wisconsin. By jointly analyzing these 34 isolates with 26 isolates previously reported, we determined that geographic barriers may limit S. neurona dispersal and that only a limited subset of possible parasite genotypes may have been introduced to recently established opossum populations. Moreover, our study confirms that diverse intermediate hosts share a common infection source, the opossum (Didelphis virginiana).

  9. Characterization of Sarcocystis from four species of hawks from Georgia, USA.

    Science.gov (United States)

    Yabsley, Michael J; Ellis, Angela E; Stallknecht, David E; Howerth, Elizabeth W

    2009-02-01

    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.

  10. Sarcocystis neurona: molecular characterization of enolase domain I region and a comparison to other protozoa.

    Science.gov (United States)

    Bolten, K E; Marsh, A E; Reed, S M; Dubey, J P; Toribio, R E; Saville, W J A

    2008-09-01

    Sarcocystis neurona causes protozoal myeloencephalitis and has the ability to infect a wide host range in contrast to other Sarcocystis species. In the current study, five S. neurona isolates from a variety of sources, three Sarcocystis falcatula, one Sarcocystis dasypi/S. neurona-like isolate, and one Besnoitia darlingi isolate were used to compare the enolase 2 gene segment containing the domain I region to previously sequenced enolase genes from Neospora caninum, Neospora hughesi, Toxoplasma gondii, Plasmodium falciparum, and Trypanosoma cruzi; enolase 2 segment containing domain I region is highly conserved amongst these parasites of veterinary and medical importance. Immunohistochemistry results indicates reactivity of T. gondii enolase 1 and 2 antibodies to S. neurona merozoites and metrocytes, but no reactivity of anti-enolase 1 to the S. neurona bradyzoite stage despite reactivity to T. gondii bradyzoites, suggesting expression differences between organisms.

  11. Molecular typing of Sarcocystis neurona: current status and future trends.

    Science.gov (United States)

    Elsheikha, Hany M; Mansfield, Linda S

    2007-10-21

    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.

  12. First Molecular Identification of Sarcocystis ovicanis (Protozoa, Apicomplexa in the Brain of Sheep in Iran.

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    Mitra Salehi

    2014-06-01

    Full Text Available The objective of the present study was to survey the presence of Sarcocystis in sheep's brain in North Khorasan Province.In general, 80 samples of sheep's brain were collected from slaughtered sheep in slaughterhouses of North Khorasan Province. Tissue digestion method was used for observing bradyzoites in tissues. Histopathological processing tracing Sarcocystis and ensuing structural change in the brain tissue were conducted. PCR analysis was conducted on all the brain samples. Sequencing was done for one PCR product. Genotype was identified by Blast search and homology analysis.Sarcocystis spp. was found in one of the brain samples (1.25% using tissue digestion method. The presence of bradyzoite was also confirmed in the prepared histopathological sections. PCR analysis was positive in one of samples. Genotyping of one sample proved that Sarcocystis species was Sarcocystis ovicanis and the nucleotide sequence of this parasite was deposited in the GenBank database under accession number No.KF489431.Sarcocystis ovicanis can involve brain tissue of sheep and consequently causes clinical symptoms.

  13. Early migration of Sarcocystis neurona in ponies fed sporocysts.

    Science.gov (United States)

    Elitsur, E; Marsh, A E; Reed, S M; Dubey, J P; Oglesbee, M J; Murphy, J E; Saville, W J A

    2007-10-01

    Sarcocystis neurona is the most important cause of equine protozoal myeloencephalitis (EPM), a neurologic disease of the horse. In the present work, the kinetics of S. neurona invasion is determined in the equine model. Six ponies were orally inoculated with 250 x 10(6) S. neurona sporocysts via nasogastric intubation and killed on days 1, 2, 3, 5, 7, and 9 postinoculation (PI). At necropsy, tissue samples were examined for S. neurona infection. The parasite was isolated from the mesenteric lymph nodes at 1, 2, and 7 days PI; the liver at 2, 5, and 7 days PI; and the lungs at 5, 7, and 9 days PI by bioassays in interferon gamma gene knock out mice (KO) and from cell culture. Microscopic lesions consistent with an EPM infection were observed in brain and spinal cord of ponies killed 7 and 9 days PI. Results suggest that S. neurona disseminates quickly in tissue of naive ponies.

  14. Penetration of equine leukocytes by merozoites of Sarcocystis neurona.

    Science.gov (United States)

    Lindsay, David S; Mitchell, Sheila M; Yang, Jibing; Dubey, J P; Gogal, Robert M; Witonsky, Sharon G

    2006-06-15

    Horses are considered accidental hosts for Sarcocystis neurona and they often develop severe neurological disease when infected with this parasite. Schizont stages develop in the central nervous system (CNS) and cause the neurological lesions associated with equine protozoal myeloencephalitis. The present study was done to examine the ability of S. neurona merozoites to penetrate and develop in equine peripheral blood leukocytes. These infected host cells might serve as a possible transport mechanism into the CNS. S. neurona merozoites penetrated equine leukocytes within 5 min of co-culture. Infected leukocytes were usually monocytes. Infected leukocytes were present up to the final day of examination at 3 days. Up to three merozoites were present in an infected monocyte. No development to schizont stages was observed. All stages observed were in the host cell cytoplasm. We postulate that S. neurona merozoites may cross the blood brain barrier hidden inside leukocytes. Once inside the CNS these merozoites can egress and invade additional cells and cause encephalitis.

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

    DEFF Research Database (Denmark)

    Dahlgren, S.S.; Oliveira, Rodrigo Gouveia; Gjerde, B.

    2008-01-01

    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...

  16. Lack of Sarcocystis neurona antibody response in Virginia opossums (Didelphis virginiana) fed Sarcocystis neurona-infected muscle tissue.

    Science.gov (United States)

    Cheadle, M A; Lindsay, D S; Greiner, E C

    2006-06-01

    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.

  17. A review of Sarcocystis of domestic animals and of other coccidia of cats and dogs.

    Science.gov (United States)

    Dubey, J P

    1976-11-15

    The nomenclature, life cycles, and pathogenicity of Sarcocystis of domestic animals are reviewed. Sarcocystis had a 2-host life cycle, with carnivores as definitive hosts and herbivores as intermediate hosts. The following species are found in domestic animals (with the definitive hosts given in parentheses): 3 species in the ox: S cruzi (dog, wolf, coyote, raccoon, fox), S hirsuta (cat), S hominis (man, monkey); 2 species in the sheep: S ovicanis (dog), S tenella (cat); 3 species in the pig: S miescheriana (dog), S porcifelis n sp (cat), S porcihominis n sp (man); and 1 species in the horse: S bertrami (dog). Sarcocystis cruzi, S ovicanis, and S porcifelis are highly pathogenic to the ox, the sheep, and the pig, respectively. Clinical signs of acute bovine sarcocystosis are: anorexia, pyrexia (42 C, or more), anemia, cachexia, enlarged palpable lymph nodes, excessive salivation, and loss of hair at the tip of the tail. Anemia, anorexia, ataxia, and abortions are the chief clinical signs of acute ovine sarcocystosis. These signs are evident at the time of vascular endothelium is parasitized by schizonts. The schizonts disappear in about 1 month, and cysts are formed in the muscles. The cystic phase of sarcocystosis is virtually nonpathogenic. Carnivores shed sporocysts in their feces after ingesting the intramuscular cysts from the herbivores. Sarcocystis is nonpathogenic to the definitive host. Feline and canine coccidia are also reviewed. The following 11 species are found in cats: Toxoplasma gondii, Hammondia hammondi, Isospora felis, Isosporarivolta, Besnoitia besnoiti, Besnoitia sp, and 5 types of Sarcocystis (S hirsuta from the ox, S tenella from the sheep, S muris from the mouse, S porcifelis from the pig, and Sarcocystis sp from Grant's gazelle). The following 10 species are found in canine feces (Isospora canis, Isospora ohioensis, Isospora wallacei n sp; and 7 types of Sarcocystis (S cruzi from the ox, S ovicanis from the sheep, S bertrami and Sarcocystis

  18. Parasitemia due to Sarcocystis neurona-like infection in a clinically ill domestic cat.

    Science.gov (United States)

    Zitzer, Nina C; Marsh, Antoinette E; Burkhard, Mary Jo; Radin, M Judith; Wellman, Maxey L; Jugan, Maria; Parker, Valerie

    2017-09-11

    An 8-year-old, 6-kg, male neutered Domestic Shorthair cat was presented to The Ohio State University Veterinary Medical Center (OSU-VMC) for difficulty breathing. Physical examination and thoracic radiographs indicated pneumonia, a soft-tissue mass in the left caudal lung lobe, and diffuse pleural effusion. The effusion was classified as modified transudate. Rare extracellular elongated (~5-7 μm × 1-2 μm) zoites with a central round to oval-shaped purple to deep purple vesicular nucleus with coarsely stippled chromatin and light blue cytoplasm were seen on a peripheral blood smear. Serum IgG and IgM were positive for Sarcocystis sp. antibodies and negative for Toxoplasma gondii antibodies, suggesting that the infection was acute rather than a recrudescence of prior infection. This organism was most consistent with either Sarcocystis neurona or Sarcocystis dasypi based on DNA sequence analysis of PCR products using COC ssRNA, ITS-1, snSAG2, and JNB25/JD396 primer sets. This is the first report to visualize by light microscopy circulating Sarcocystis sp. merozoites in the peripheral blood of a domestic cat. Therefore, Sarcocystis should be considered as a differential diagnosis in cats with suspected systemic protozoal infection. © 2017 American Society for Veterinary Clinical Pathology.

  19. Sarcocystis cruzi (Apicomplexa: Sarcocystidae) no cachorro-do-mato (Cerdocyon thous)

    OpenAIRE

    Janaina S. Rodrigues; Gisele S. Meireles; Paulo R. Carvalho Filho; Ribeiro, Carlos T.; Flausino,Walter; Lopes,Carlos Wilson G.

    2008-01-01

    Esporocistos de Sarcocystis foram identificados nas amostras fecais de um cachorro-do-mato. Eles foram dados por via oral para um bezerro em aleitamento, sendo observados cistos com morfologia compatível com os de Sarcocystis cruzi na musculatura cardíaca e esquelética, três meses após a infecção. Musculatura cardíaca deste bezerro foi dada para um segundo cão doméstico livre de coccídios, que eliminou esporocistos compatíveis com os de Sarcocystis em suas fezes, tendo com períodos pré-patent...

  20. Sarcocystis canis n. sp. (Apicomplexa: Sarcocystidae), the etiologic agent of generalized coccidiosis in dogs.

    Science.gov (United States)

    Dubey, J P; Speer, C A

    1991-08-01

    Sarcocystis canis n. sp. is proposed for the protozoon associated with encephalitis, hepatitis, and generalized coccidiosis in dogs. Only asexual stages are known in macrophages, neurons, dermal, and other cells of the body. The parasite is located free in the host cell cytoplasm without a parasitophorous vacuole; schizonts divide by endopolygeny. Schizonts are 5-25 x 4-20 microns and contain 6-40 merozoites. Merozoites are approximately 5-7 microns x 1 micron and do not contain rhoptries. The parasite is PAS-negative and reacts with Sarcocystis cruzi antiserum but not with Toxoplasma gondii, Neospora caninum, or Caryospora bigenetica antisera in an immunohistochemical test.

  1. Aborto ovino associado com infecção por Sarcocystis sp Ovine abortion associated with Sarcocystis sp. infection

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    Caroline A. Pescador

    2007-10-01

    Full Text Available Infecções por protozoários têm distribuição mundial e podem causar aborto, nascimentos prematuros e ou morte fetal em diversas espécies animais. Em julho de 2004, oito ovinos Corriedale apresentaram problemas reprodutivos caracterizados por aborto e natimortalidade no terço final da gestação. Dessas oito perdas, um natimorto macho foi enviado ao Setor de Patologia Veterinária para necropsia. Alterações macroscópicas não foram observadas durante a necropsia. Lesões histológicas foram observadas principalmente no cérebro e coração e se caracterizaram por encefalite não-supurativa multifocal acentuada associada à presença de protozoários no interior de células endoteliais e vasos sanguíneos e miocardite não-supurativa focal leve. Alguns desses organismos apresentaram formato de roseta. O teste de imunoistoquímica anti-Toxoplasma gondii foi negativo, mas houve reação cruzada com anticorpo anti-Neospora caninum. O exame de imunofluorescência direta para Leptospira sp. foi negativo. A bacteriologia aeróbica e micro-aeróbica não revelou crescimento significativo. Esses achados foram compatíveis com o diagnóstico de Sarcocystis sp.Protozoal infection has worldwide distribution and may cause abortion, premature parturition or fetal death in almost all domestic animals. In July 2004, eight Corriedale sheep showed abortion and stillbirth in the third trimester of gestation. Of these reproductive losses, one stillborn male was submitted to the Laboratory of Veterinary Pathology for necropsy investigation. The direct immunofluorescence test for Leptospira sp. was negative. No significant bacteria was isolated from lung and liver by aerobic and microaerobic cultures. Macroscopic lesions were not found in any fetal tissue. The histological lesions were observed mainly in the brain and heart and consisted primarily of severe multifocal nonsupurative encephalitis and nonsuppurative myocarditis. Schizonts of a protozoan

  2. Sarcocystis neurona retinochoroiditis in a sea otter (Enhydra lutris kenyoni)

    Science.gov (United States)

    Dubey, J.P.; Thomas, N.J.

    2011-01-01

    Sarcocystis neurona is an important cause of fatal disease in sea otters in the USA. Encephalitis is the predominant lesion and parasites are confined to the central nervous system and muscles. Here we report retinochoroiditis in a sea otter (Enhydra lutris kenyoni) found dead on Copalis Beach, WA, USA. Salient lesions were confined to the brain and eye. Multifocal nonsuppurative meningoencephalitis was present in the cerebrum and cerebellum associated with S. neurona schizonts. The retina of one eye had a focus of inflammation that contained numerous S. neurona schizonts and merozoites. The focus extended from the retinal pigment epithelium inward through all layers of the retina, but inflammation was most concentrated at the inner surface of the tapetum and the outer retina. The inner and outer nuclear layers of the retina were disorganized and irregular at the site of inflammation. There was severe congestion and mild hemorrhage in the choroid, and mild hemorrhage into the vitreous body. Immunohistochemistry with S. neurona-specific polyclonal rabbit antibodies stained schizonts and merozoites. To our knowledge this is the first report of S. neurona-associated retinochoroiditis in any naturally infected animal.

  3. Sarcocystis neurona retinochoroiditis in a sea otter (Enhydra lutris kenyoni)

    Science.gov (United States)

    Dubey, J.P.; Thomas, N.J.

    2011-01-01

    Sarcocystis neurona is an important cause of fatal disease in sea otters in the USA. Encephalitis is the predominant lesion and parasites are confined to the central nervous system and muscles. Here we report retinochoroiditis in a sea otter (Enhydra lutris kenyoni) found dead on Copalis Beach, WA, USA. Salient lesions were confined to the brain and eye. Multifocal nonsuppurative meningoencephalitis was present in the cerebrum and cerebellum associated with S. neurona schizonts. The retina of one eye had a focus of inflammation that contained numerous S. neurona schizonts and merozoites. The focus extended from the retinal pigment epithelium inward through all layers of the retina, but inflammation was most concentrated at the inner surface of the tapetum and the outer retina. The inner and outer nuclear layers of the retina were disorganized and irregular at the site of inflammation. There was severe congestion and mild hemorrhage in the choroid, and mild hemorrhage into the vitreous body. Immunohistochemistry with S. neurona-specific polyclonal rabbit antibodies stained schizonts and merozoites. To our knowledge this is the first report of S. neurona-associated retinochoroiditis in any naturally infected animal. ?? 2011 Elsevier B.V.

  4. Efficacy of decoquinate against Sarcocystis neurona in cell cultures.

    Science.gov (United States)

    Lindsay, David S; Nazir, M Mudasser; Maqbool, Azhar; Ellison, Siobhan P; Strobl, Jeannine S

    2013-09-01

    Decoquinate is a quinolone anticoccidial approved for use in the prevention of intestinal coccidiosis in farm animals. This compound has good activity against Toxoplasma gondii and Neospora caninum in cell cultures. The drug acts on the parasites' mitochondria. The activity of decoquinate against developing merozoites of 2 isolates of Sarcocystis neurona was examined in cell culture. Merozoite production at 10 days was completely inhibited when decoquinate was used at 20 or 240 nM. The IC50 of decoquinate was 0.5 ± 0.09nM for the Sn6 isolate of S. neurona from a horse and 1.1 ± 0.6 nM for the SnOP15 isolate of S. neurona from an opossum. Levamisole was toxic at 5 μg/ml and no synergism was observed when decoquinate was combined with levamisole and tested against the Sn3YFP isolate of S. neurona. Decoquinate was cidal for developing schizonts of S. neurona at 240 nM. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Molecular genetic transfection of the coccidian parasite Sarcocystis neurona.

    Science.gov (United States)

    Gaji, Rajshekhar Y; Zhang, Deqing; Breathnach, Cormac C; Vaishnava, Shipra; Striepen, Boris; Howe, Daniel K

    2006-11-01

    Sarcocystis neurona is an apicomplexan parasite that is the major cause of equine protozoal myeloencephalitis (EPM). The biology of this pathogen remains poorly understood in part due to unavailability of molecular genetic tools. Hence, with an objective to develop DNA transfection capabilities for S. neurona, the 5' flanking region of the SnSAG1 gene was isolated from a genomic library and used to construct expression plasmids. In transient assays, the reporter molecules beta-galactosidase (beta-gal) and yellow fluorescent protein (YFP) could be detected in electroporated S. neurona, thereby confirming the feasibility of transgene expression in this organism. Stable transformation of S. neurona was achieved using a mutant dihydrofolate reductase thymidylate synthase (DHFR-TS) gene of Toxoplasma gondii that confers resistance to pyrimethamine. This selection system was used to create transgenic S. neurona that stably express beta-gal and YFP. As shown in this study, these transgenic clones can be useful for analyzing growth rate of parasites in vitro and for assessing drug sensitivities. More importantly, the DNA transfection methods described herein should greatly facilitate studies examining intracellular parasitism by this important coccidian pathogen.

  6. Sarcocystis neurona retinochoroiditis in a sea otter (Enhydra lutris kenyoni).

    Science.gov (United States)

    Dubey, J P; Thomas, N J

    2011-12-29

    Sarcocystis neurona is an important cause of fatal disease in sea otters in the USA. Encephalitis is the predominant lesion and parasites are confined to the central nervous system and muscles. Here we report retinochoroiditis in a sea otter (Enhydra lutris kenyoni) found dead on Copalis Beach, WA, USA. Salient lesions were confined to the brain and eye. Multifocal nonsuppurative meningoencephalitis was present in the cerebrum and cerebellum associated with S. neurona schizonts. The retina of one eye had a focus of inflammation that contained numerous S. neurona schizonts and merozoites. The focus extended from the retinal pigment epithelium inward through all layers of the retina, but inflammation was most concentrated at the inner surface of the tapetum and the outer retina. The inner and outer nuclear layers of the retina were disorganized and irregular at the site of inflammation. There was severe congestion and mild hemorrhage in the choroid, and mild hemorrhage into the vitreous body. Immunohistochemistry with S. neurona-specific polyclonal rabbit antibodies stained schizonts and merozoites. To our knowledge this is the first report of S. neurona-associated retinochoroiditis in any naturally infected animal. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. Sarcocystis spp. in sheep and goats: frequency of infection and species identification by morphological, ultrastructural, and molecular tests in Bahia, Brazil.

    Science.gov (United States)

    Bittencourt, Marta Vasconcelos; Meneses, Iris Daniela S; Ribeiro-Andrade, Müller; de Jesus, Rogério Fernando; de Araújo, Flábio Ribeiro; Gondim, Luís F Pita

    2016-04-01

    Sarcocystis spp. are cyst-forming coccidia that infect numerous animals species, including several livestock species. Despite the importance of sheep and goat production in Brazil, little it is known about the Sarcocystis species that infect small ruminants in the country and their potential impact on meat condemnation due to the presence of macroscopic cysts of the parasite. The aims of the present study were to determine the frequency of infection by Sarcocystis spp. in goats and sheep intended for human consumption in Bahia State, Brazil, as well as to identify the parasite species in selected samples. The entire tongue, esophagus, and heart were collected from 120 goats and 120 sheep. Tissues were examined for Sarcocystis spp. by macroscopic evaluation, light microscopy, electron microscopy, and molecular tests. Microscopic cysts of Sarcocystis spp. were detected in 95.8 % of sheep and 91.6 % of goats. Using either transmission electron microscopy or partial sequencing of the 18S region of the ribosomal DNA (rDNA) for species identification, Sarcocystis tenella and Sarcocystis arieticanis were observed in sheep and Sarcocystis capracanis in goats. Macroscopic cysts were not detected in the analyzed samples. We concluded that goats and sheep destined for human consumption in Bahia possess high frequencies of Sarcocystis infection. Carcass condemnation due to Sarcocystis macrocysts seems to be rare in the studied region. S. arieticanis and S. capracanis were confirmed for the first time by electron microscopy or by molecular tests in small ruminants from Brazil.

  8. Detection of antibodies against Sarcocystis neurona, neospora spp., and Toxoplasma gondii in horses from Costa Rica

    Science.gov (United States)

    Serum samples from 315 horses from Costa Rica, Central America were examined for the presence of antibodies against Sarcocystis neurona, Neospora spp., and Toxoplasma gondii using the SnSAG2 ELISA, the NhSAG1 ELISA, and the modified agglutination test, respectively. Anti-S. neurona antibodies were f...

  9. Prevalence of antibodies to Sarcocystis neurona and Neospora hughesi in horses from Mexico

    Science.gov (United States)

    The risk of equine protozoal myeloencephalitis (EPM) to horses in Mexico has not been established. Serum samples from 495 horses in Durango State, Mexico were examined for the presence of antibodies to Sarcocystis neurona and Neospora hughesi using enzyme-linked immunosorbent assays (ELISAs) based o...

  10. Bobcat (Lynx rufus) as a new natural intermediate host for Sarcocystis neurona

    Science.gov (United States)

    The protozoan Sarcocystis neurona is an important cause of severe clinical disease of horses (called equine protozoal myeloencephalitis, EPM), marine mammals, companion animals, and several species of wildlife animals in the Americas. The Virginia opossum (Didelphis virginiana) is its definitive hos...

  11. Acute, fatal Sarcocystis calchasi-associated hepatitis in Roller pigeons (Columbia livia f. dom.) at Philadelphia Zoo

    Science.gov (United States)

    Four Roller pigeons (Columba livia f. dom.) at the Philadelphia Zoo died suddenly. Necropsy examination revealed macroscopic hepatitis. Microscopically, the predominant lesions were in liver, characterized with necrosis and mixed cell inflammatory response. Sarcocystis calchasi-like schizonts and fr...

  12. Study of Zoonotic Tissue Parasites (Hydatid Cyst, Fasciola, Dicrocoelium and Sarcocystis in Hamadan Abattoir

    Directory of Open Access Journals (Sweden)

    M. Fallah

    2010-10-01

    Full Text Available Introduction & Objectives: Zoonotic parasites are large groups of zoonoses among which the most important are hydatid cyst, liver trematodes and sarcocystis.These zoonoses are of considerable importance regarding both human health and economy. The objective of this study was to determine the prevalence of tissue zoonotic parasites and their epidemiologic status in Hamadan and to estimate the health and medical burden they impose on the society.Materials & Methods: In this cross sectional study, viscera (including liver, lung, kidney, heart,… and muscles of 2590 sheep, 420 cattle, and 490 goats were macroscopically inspected for hydatid cysts, liver flukes, cysticercus , and microscopically (for Sarcocystis in the Hamadan abattoir. The data were presented by descriptive tables and analyzed by 2 statistical test. Results: The infection rate for hydatid cyst, Fasciola, Dicrocoelium and Sarcocystis were found 12.3%, 4.9%, 6.5%, and 5.5% respectively. The high infection rates for hydatid cyst and Fasciola were found in cattle (16.2% and 9.5% and for Dicrocoelium and Sarcocystis were found in sheep (6.9%. Infection rate of lungs was higher (41.2% than liver (36.6% and liver and lung simultaneously were 22.2% in the infected animals. Infection to Sarcocystis and Cysticercus were not found in the cattle. Conclusion: This study indicated that infection rate of tissue zoonotic parasites are relatively high in the domestic animals of Hamadan , however, the rate is lower in comparison to the previous studies. These parasites had imposed considerable economic burden on the society through reduction in the dairy production and increased the risk of infection in the population as well. (Sci J Hamadan Univ Med Sci 2010;17(3: 5-12

  13. Biological characterisation of Sarcocystis neurona isolated from a Southern sea otter (Enhydra lutris nereis)

    Science.gov (United States)

    Lindsay, D.S.; Thomas, N.J.; Dubey, J.P.

    2000-01-01

    Sarcocystis neurona was isolated from the brain of a juvenile, male southern sea otter (Enhydra lutris nereis) suffering from CNS disease. Schizonts and merozoites in tissue sections of the otter's brain reacted with anti-S. neurona antiserum immunohistochemically. Development in cell culture was by endopolyogeny and mature schizonts were first observed at 3 days postinoculation. PCR of merozoite DNA using primer pairs JNB33/JNB54 and restriction enzyme digestion of the 1100 bp product with Dra I indicated the organism was S. neurona. Four of four interferon-γ gene knockout mice inoculated with merozoites developed S. neurona-associated encephalitis. Antibodies to S. neurona but not Sarcocystis falcatula, Toxoplasma gondii, or Neospora caninum were present in the serum of inoculated mice. This is the first isolation of S. neurona from the brain of a non-equine host.

  14. Identification and intraspecific genetic diversity of Sarcocystis rileyi from ducks, Anas spp., in Lithuania and Finland.

    Science.gov (United States)

    Prakas, P; Oksanen, A; Butkauskas, D; Sruoga, A; Kutkienė, L; Švažas, S; Isomursu, M; Liaugaudaitė, S

    2014-10-01

    Macroscopic Sarcocystis cysts were detected in the muscles of 28 Mallards ( Anas platyrhynchos ), 1 Eurasian Wigeon ( Anas penelope ), and 1 Common Teal ( Anas crecca ) hunted in Lithuania and Finland. According to the sequences of the 18S rRNA gene, 28S rRNA gene, and ITS-1 region, the macrocysts examined from all 30 ducks belonged to Sarcocystis rileyi. This parasite was found in the Eurasian Wigeon and the Common Teal for the first time. All S. rileyi isolates examined were identical to each other and differed from 2 S. rileyi isolates previously reported from 2 Mallards from the United States only by 1 nucleotide substitution within the ITS-1 region.

  15. Pathology produced by, prevalence, of, and probable life-cycle of a species of Sarcocystis in the domestic fowl.

    Science.gov (United States)

    Munday, B L; Humphrey, J D; Kila, V

    1977-01-01

    Sarcosporidiosis was found to be the cause of a severe myositis in 3 fowls in Papua New Guinea and 2 in Australia. This represented 3.8% of a series of fowls examined in Papua New Guinea. The overall prevalence of infection in these birds was 45%. Both epidemiological and experimental evidence suggested that the dog was the definitive host for this particular type of sarcocyst.

  16. DNA extraction methods and multiple sampling to improve molecular diagnosis of Sarcocystis spp. in cattle hearts.

    Science.gov (United States)

    Bräunig, Patrícia; Portella, Luiza Pires; Cezar, Alfredo Skrebsky; Libardoni, Felipe; Sangioni, Luis Antonio; Vogel, Fernanda Silveira Flores; Gonçalves, Paulo Bayard Dias

    2016-10-01

    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.

  17. Necrotizing meningoencephalitis caused by Sarcocystis falcatula in bare-faced ibis (Phimosus infuscatus).

    Science.gov (United States)

    Konradt, Guilherme; Bianchi, Matheus Viezzer; Leite-Filho, Ronaldo Viana; da Silva, Bruna Zafalon; Soares, Rodrigo Martins; Pavarini, Saulo Petinatti; Driemeier, David

    2017-02-01

    The infection by S. falcatula is commonly associated with respiratory disease in captive psittacine birds, with a few case reports of this protozoan causing encephalitis in wild birds. We describe the clinical, pathological, and molecular aspects of an infection by S. falcatula in a bare-faced ibis (Phimosus infuscatus). Clinically, wing paralysis and mild motor incoordination were observed. At necropsy, the telencephalic cortex showed multifocal to coalescing yellowish soft areas. Histologically, multifocal to coalescent nonsuppurative necrotizing meningoencephalitis of telencephalic cortex, cerebellum, and brainstem was observed. Necrotic areas showed multiple protozoan organism characteristics of Sarcocystis sp. schizonts in the cytoplasm of endothelial cells or lying free in the neuropil. Partial genetic sequences of the gene encoding cytochrome b (CYTB), the gene encoding the beta subunit of RNA polymerase (RPOB) and the first internal transcribed spacer (ITS-1) from Sarcocystis sp. schizonts revealed that the parasite had ITS-1 sequences that were 100% identical to the homologous alleles from Sarcocystis sp. shed by Didelphis albiventris in Brazil. RPOB and CYTB sequences were 100% identical to homologous of S. falcatula available in Genbank. Thus, this is the first report of necrotizing meningoencephalitis caused by S. falcatula in bare-faced ibis (P. infuscatus).

  18. Systems-based analysis of the Sarcocystis neurona genome identifies pathways that contribute to a heteroxenous life cycle.

    Science.gov (United States)

    Blazejewski, Tomasz; Nursimulu, Nirvana; Pszenny, Viviana; Dangoudoubiyam, Sriveny; Namasivayam, Sivaranjani; Chiasson, Melissa A; Chessman, Kyle; Tonkin, Michelle; Swapna, Lakshmipuram S; Hung, Stacy S; Bridgers, Joshua; Ricklefs, Stacy M; Boulanger, Martin J; Dubey, Jitender P; Porcella, Stephen F; Kissinger, Jessica C; Howe, Daniel K; Grigg, Michael E; Parkinson, John

    2015-02-10

    Sarcocystis neurona is a member of the coccidia, a clade of single-celled parasites of medical and veterinary importance including Eimeria, Sarcocystis, Neospora, and Toxoplasma. Unlike Eimeria, a single-host enteric pathogen, Sarcocystis, Neospora, and Toxoplasma are two-host parasites that infect and produce infectious tissue cysts in a wide range of intermediate hosts. As a genus, Sarcocystis is one of the most successful protozoan parasites; all vertebrates, including birds, reptiles, fish, and mammals are hosts to at least one Sarcocystis species. Here we sequenced Sarcocystis neurona, the causal agent of fatal equine protozoal myeloencephalitis. The S. neurona genome is 127 Mbp, more than twice the size of other sequenced coccidian genomes. Comparative analyses identified conservation of the invasion machinery among the coccidia. However, many dense-granule and rhoptry kinase genes, responsible for altering host effector pathways in Toxoplasma and Neospora, are absent from S. neurona. Further, S. neurona has a divergent repertoire of SRS proteins, previously implicated in tissue cyst formation in Toxoplasma. Systems-based analyses identified a series of metabolic innovations, including the ability to exploit alternative sources of energy. Finally, we present an S. neurona model detailing conserved molecular innovations that promote the transition from a purely enteric lifestyle (Eimeria) to a heteroxenous parasite capable of infecting a wide range of intermediate hosts. Sarcocystis neurona is a member of the coccidia, a clade of single-celled apicomplexan parasites responsible for major economic and health care burdens worldwide. A cousin of Plasmodium, Cryptosporidium, Theileria, and Eimeria, Sarcocystis is one of the most successful parasite genera; it is capable of infecting all vertebrates (fish, reptiles, birds, and mammals-including humans). The past decade has witnessed an increasing number of human outbreaks of clinical significance associated with

  19. Fatal hepatic sarcocystosis in a captive black bear (Ursus americanus) associated with Sarcocystis canis-like infection.

    Science.gov (United States)

    Davies, Jennifer L; Haldorson, Gary J; Bradway, Dan S; Britton, Ann P

    2011-03-01

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

  20. Molecular identification of Sarcocystis rileyi sporocysts in red foxes (Vulpes vulpes) and raccoon dogs (Nyctereutes procyonoides) in Lithuania.

    Science.gov (United States)

    Prakas, Petras; Liaugaudaitė, Simona; Kutkienė, Liuda; Sruoga, Aniolas; Švažas, Saulius

    2015-05-01

    Despite the fact that Sarcocystis rileyi is one of the earliest described species of the genus Sarcocystis forming macrocysts in ducks, the life cycle of this species is still unknown in Europe. Sarcocystis spp. oocysts/sporocysts were observed in faeces of four of 23 (17.4 %) and in small intestine mucosal scrapings of four of 20 (20.0 %) red foxes (Vulpes vulpes) and in small intestine mucosal scrapings of seven of 13 (53.8 %) raccoon dogs (Nyctereutes procyonoides) hunted in Lithuania. A very small number of Sarcocystis sporocysts measuring 11.9 × 8.3 μm (n = 5) was found in faecal samples, whereas considerably more sporulated Sarcocystis oocysts and free sporocysts were detected in the small intestines of red foxes and raccoon dogs. These sporocysts measured 12.9 × 8.1 μm (n = 16) and 12.1 × 8.1 μm (n = 54) in red foxes and raccoon dogs, respectively. Using species-specific PCR and subsequent sequencing, internal transcribed spacer 1 (ITS-1) region partial sequences of oocysts/sporocysts from small intestine mucosal scrapings of six raccoon dogs and three red foxes were identified as belonging to S. rileyi. The present study provides strong evidence showing that the red fox and the raccoon dog can serve as final hosts of S. rileyi in Europe; however, transmission experiments are needed for the ultimate approval.

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

    Science.gov (United States)

    Rooney, A L; Limon, G; Vides, H; Cortez, A; Guitian, J

    2014-10-01

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

  2. Molecular Analysis of Sarcocystis Spp. Isolated from Sheep (Ovis aries in Babol Area, Mazandaran Province, Northern Iran

    Directory of Open Access Journals (Sweden)

    Narges KALANTARI

    2016-03-01

    Full Text Available Background: To differentiate Sarcocystis macro-cyst-forming species in slaughtered sheep in Babol area, Mazandaran Province, sequence analysis of 18S rRNA gene was performed.Methods: Overall, 150 slaughtered sheep were examined macroscopically in slaughterhouse, Babol and intra-abdominal and diaphragm muscles tissues infected with macro-cyst of Sarcocystis spp. were collected in 2013. One macro-cyst was isolated from the infected muscles of each sheep. The partial 18S rRNA gene was amplified by PCR and sequenced afterward.Results: The rate of infection with macro-cyst producing Sarcocystis spp. was 33.3% (50 / 150. The partial 18S rRNA gene of Sarcocystis species was amplified at the expected PCR product size (~1100 bp from all 50 macroscopic cysts samples. From 30 sequences DNA samples, 20 samples (66.7%, six (20% and four (13.3% isolates were identified as S. gigantea, S. moulei and Sarcocystis spp., respectively. Eight and thirty-four variations in nucleotide position were seen in partial sequence of the18S rRNA gene of S. gigantea and S. moulei.Conclusion: Sheep can be considered as an alternative intermediate host for S. moulei. Furthermore, multiple alignments showed some variations in the consensus sequences of the isolates obtained in the current study compared with previously published isolates. To understand better the genetic diversity among Sarcocystis species complete sequences of the18S rRNA gene or sequence analysis of other genetic loci would be beneficial.

  3. Rhinitis and disseminated disease in a ferret (Mustela putorius furo) naturally infected with Sarcocystis neurona.

    Science.gov (United States)

    Britton, Ann P; Dubey, J P; Rosenthal, Benjamin M

    2010-04-19

    Naturally occurring Sarcocystis neurona infection in a ferret (Mustela putorius furo) with rhinitis and disseminated disease are described for the first time. The ferret exhibited severe rhinitis with intra-lesional S. neurona merozoites and schizonts. Diagnosis was confirmed immunohistochemically by staining with S. neurona-specific antibodies, and by phylogenetic analyses of conserved and variable portions of nuclear ribosomal DNA. On the basis of intense schizogony in the nasal mucosa, we propose the possibility of an olfactory nerve pathway route of infection for S. neurona meningoencephalitis.

  4. An update on Sarcocystis neurona infections in animals and equine protozoal myeloencephalitis (EPM).

    Science.gov (United States)

    Dubey, J P; Howe, D K; Furr, M; Saville, W J; Marsh, A E; Reed, S M; Grigg, M E

    2015-04-15

    Equine protozoal myeloencephalitis (EPM) is a serious disease of horses, and its management continues to be a challenge for veterinarians. The protozoan Sarcocystis neurona is most commonly associated with EPM. S. neurona has emerged as a common cause of mortality in marine mammals, especially sea otters (Enhydra lutris). EPM-like illness has also been recorded in several other mammals, including domestic dogs and cats. This paper updates S. neurona and EPM information from the last 15 years on the advances regarding life cycle, molecular biology, epidemiology, clinical signs, diagnosis, treatment and control. Published by Elsevier B.V.

  5. Prevalence of antibodies to Trypanosoma cruzi, Toxoplasma gondii, Encephalitozonn cuniculi, Sarcocystis neurona, Besnoitia darlingi, and Neospora caninum in North American opossum, Didelphis virginiana, from Southern Louisian

    Science.gov (United States)

    We examined the prevalence of antibodies to zoonotic protozoan parasites (Trypanosoma cruzi, Toxoplasma gondii, and Encephalitozoon cuniculi) and protozoan’s of veterinary importance (Neospora caninum, Sarcocystis neurona and Besnoitia darlingi) in a population of North American opossums (Didelphis...

  6. Pervasive Environmental Contamination with Human Feces Results in High Prevalence of Zoonotic Sarcocystis Infection in Pigs in the Punjab, India.

    Science.gov (United States)

    Kaur, M; Singh, B B; Sharma, R; Gill, J P S

    2016-04-01

    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.

  7. Effects of Experimental Sarcocystis neurona-Induced Infection on Immunity in an Equine Model

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    S. Rochelle Lewis

    2014-01-01

    Full Text Available Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM, affecting 0.5–1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both.

  8. Enzyme-Linked Immunosorbent Assays for Detection of Equine Antibodies Specific to Sarcocystis neurona Surface Antigens†

    Science.gov (United States)

    Hoane, Jessica S.; Morrow, Jennifer K.; Saville, William J.; Dubey, J. P.; Granstrom, David E.; Howe, Daniel K.

    2005-01-01

    Sarcocystis neurona is the primary causative agent of equine protozoal myeloencephalitis (EPM), a common neurologic disease of horses in the Americas. We have developed a set of enzyme-linked immunosorbent assays (ELISAs) based on the four major surface antigens of S. neurona (SnSAGs) to analyze the equine antibody response to S. neurona. The SnSAG ELISAs were optimized and standardized with a sample set of 36 equine sera that had been characterized by Western blotting against total S. neurona parasite antigen, the current gold standard for S. neurona serology. The recombinant SnSAG2 (rSnSAG2) ELISA showed the highest sensitivity and specificity at 95.5% and 92.9%, respectively. In contrast, only 68.2% sensitivity and 71.4% specificity were achieved with the rSnSAG1 ELISA, indicating that this antigen may not be a reliable serological marker for analyzing antibodies against S. neurona in horses. Importantly, the ELISA antigens did not show cross-reactivity with antisera to Sarcocystis fayeri or Neospora hughesi, two other equine parasites. The accuracy and reliability exhibited by the SnSAG ELISAs suggest that these assays will be valuable tools for examining the equine immune response against S. neurona infection, which may help in understanding the pathobiology of this accidental parasite-host interaction. Moreover, with modification and further investigation, the SnSAG ELISAs have potential for use as immunodiagnostic tests to aid in the identification of horses affected by EPM. PMID:16148170

  9. Enzyme-linked immunosorbent assays for detection of equine antibodies specific to Sarcocystis neurona surface antigens.

    Science.gov (United States)

    Hoane, Jessica S; Morrow, Jennifer K; Saville, William J; Dubey, J P; Granstrom, David E; Howe, Daniel K

    2005-09-01

    Sarcocystis neurona is the primary causative agent of equine protozoal myeloencephalitis (EPM), a common neurologic disease of horses in the Americas. We have developed a set of enzyme-linked immunosorbent assays (ELISAs) based on the four major surface antigens of S. neurona (SnSAGs) to analyze the equine antibody response to S. neurona. The SnSAG ELISAs were optimized and standardized with a sample set of 36 equine sera that had been characterized by Western blotting against total S. neurona parasite antigen, the current gold standard for S. neurona serology. The recombinant SnSAG2 (rSnSAG2) ELISA showed the highest sensitivity and specificity at 95.5% and 92.9%, respectively. In contrast, only 68.2% sensitivity and 71.4% specificity were achieved with the rSnSAG1 ELISA, indicating that this antigen may not be a reliable serological marker for analyzing antibodies against S. neurona in horses. Importantly, the ELISA antigens did not show cross-reactivity with antisera to Sarcocystis fayeri or Neospora hughesi, two other equine parasites. The accuracy and reliability exhibited by the SnSAG ELISAs suggest that these assays will be valuable tools for examining the equine immune response against S. neurona infection, which may help in understanding the pathobiology of this accidental parasite-host interaction. Moreover, with modification and further investigation, the SnSAG ELISAs have potential for use as immunodiagnostic tests to aid in the identification of horses affected by EPM.

  10. Effects of Experimental Sarcocystis neurona-Induced Infection on Immunity in an Equine Model.

    Science.gov (United States)

    Lewis, S Rochelle; Ellison, Siobhan P; Dascanio, John J; Lindsay, David S; Gogal, Robert M; Werre, Stephen R; Surendran, Naveen; Breen, Meghan E; Heid, Bettina M; Andrews, Frank M; Buechner-Maxwell, Virginia A; Witonsky, Sharon G

    2014-01-01

    Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM), affecting 0.5-1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both.

  11. Antigenic evaluation of a recombinant baculovirus-expressed Sarcocystis neurona SAG1 antigen.

    Science.gov (United States)

    Gupta, G D; Lakritz, J; Saville, W J; Livingston, R S; Dubey, J P; Middleton, J R; Marsh, A E

    2004-10-01

    Sarcocystis neurona is the primary parasite associated with equine protozoal myeloencephalitis (EPM). This is a commonly diagnosed neurological disorder in the Americas that infects the central nervous system of horses. Current serologic assays utilize culture-derived parasites as antigen. This method requires large numbers of parasites to be grown in culture, which is labor intensive and time consuming. Also, a culture-derived whole-parasite preparation contains conserved antigens that could cross-react with antibodies against other Sarcocystis species and members of Sarcocystidae such as Neospora spp., Hammondia spp., and Toxoplasma gondii. Therefore, there is a need to develop an improved method for the detection of S. neurona-specific antibodies. The sera of infected horses react strongly to surface antigen 1 (SnSAG1), an approximately 29-kDa protein, in immunoblot analysis, suggesting that it is an immunodominant antigen. The SnSAG1 gene of S. neurona was cloned, and recombinant S. neurona SAG1 protein (rSnSAG1-Bac) was expressed with the use of a baculovirus system. By immunoblot analysis, the rSnSAG1-Bac antigen detected antibodies to S. neurona from naturally infected and experimentally inoculated equids, cats, rabbit, mice, and skunk. This is the first report of a baculovirus-expressed recombinant S. neurona antigen being used to detect anti-S. neurona antibodies in a variety of host species.

  12. Occurrence and Distribution of Sarcocystis Parasite Isolated from Sheep in Yazd Province, Iran

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    Bahador Hajimohammadi

    2014-11-01

    Full Text Available Introduction: Sarcocystis species are one of the most important meat borne parasites. Sarcocystosis causes several symptoms in human as well as numerous diseases with high economic impact in livestock. In current paper at first results of a pilot study for determination of disease agent in lamb of Yazd province central Iran is presented. Then status of parasite in red meat products in study region gets discussed. Materials and Methods: Muscles of 70 slaughtered Sheep from both sexes and different ages were investigated for presence of parasite cysts during September to October 2013. Carcass inspection with naked eye at industrial slaughterhouse of Yazd for macroscopic cysts, and pepsin-digestion method for microscopic cysts was performed on common infected sites of infection. Results: No macroscopic cyst was seen at inspection, however bradyzoites of parasite were observed in 97.14% of animals’ digested muscles. No significant difference between infection and age groups or sex of animals was observed. Conclusion: As humans are considered as final and intermediate host of different species of Sarcocystis, and parasite cysts present at microscopic sizes, transmission of infection from lamb meat should come into consideration. Hence public training of disease health importance, parasite cycle and methods of inactivation probable cysts in meat is recommended.

  13. Prevalence of antibodies against Neospora spp. and Sarcocystis neurona in donkeys from northeastern Brazil

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    Solange Maria Gennari

    2016-03-01

    Full Text Available Abstract Sarcocystis neurona and Neospora hughesi are coccidian protozoa that can cause neurological illness in horses in America. In this study we report seroprevalence of Neospora spp. andS. neurona in sera of 333 donkeys from the northeastern region of Brazil. Antibodies to Neospora spp. were detected in 2% (7 donkeys of 333 sera tested by the indirect fluorescent antibody test (IFAT with a cut-off dilution of 1:40. Antibodies to S. neurona were found in 3% (10 donkeys of the samples tested by IFAT (cut-off ≥50 and 21% (69 donkeys by the direct agglutination test (SAT ≥50. The SAT and IFAT results for S. neurona showed a poor concordance (value of Kappa=0.051. This is the first report ofNeospora spp. antibodies in Brazilian donkeys and the first detection of antibodies against S. neurona in this animal species.

  14. Daily feeding of diclazuril top dress pellets in foals reduces seroconversion to Sarcocystis neurona.

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    Pusterla, Nicola; Packham, Andrea; Mackie, Sarah; Kass, Philip H; Hunyadi, Laszlo; Conrad, Patricia A

    2015-11-01

    Thirty-three foals from a farm with a high exposure rate to Sarcocystis neurona were assigned to either an untreated or a diclazuril-treated group. Treated foals received daily 0.5 mg/kg of diclazuril pellets from 1 to 12 months of age. Monthly blood was tested for IgG against S. neurona using the indirect fluorescent antibody test. Following ingestion of colostral antibodies to S. neurona, there was a steady and continuous decline in seroprevalence to S. neurona until foals from both groups reached weaning age. Thereafter, the untreated foal group showed a significant increase in monthly seroprevalence compared to the diclazuril-treated foal group. The difference in temporal seroprevalence could be explained by the successful reduction of S. neurona infection in foals receiving a daily low-dose diclazuril. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Prevalence of antibodies to Sarcocystis neurona in cats from Virginia and Pennsylvania.

    Science.gov (United States)

    Hsu, Vasha; Grant, David C; Dubey, J P; Zajac, Anne M; Lindsay, David S

    2010-08-01

    Sarcocystis neurona is best known as the causative agent of equine protozoal myeloencephalitis of horses in the Americas. Domestic cats ( Felis domesticus ) were the first animals described as an intermediate host for S. neurona . However, S. neurona -associated encephalitis has also been reported in naturally infected cats in the United States. Thus, cats can be implicated in the life cycle of S. neurona as natural intermediate hosts. The present study examined the seroprevalence of IgG antibodies to merozoites of S. neurona in populations of domestic cats from Virginia and Pennsylvania. Overall, sera or plasma from 441 cats (Virginia = 232, Pennsylvania = 209) were tested by an indirect immunofluorescent assay at a 1ratio50 dilution. Antibodies to S. neurona were found in 32 (7%) of 441 cats. Of these, 22 (9%) of the 232 cats from Virginia and 10 (5%) of the 209 cats from Pennsylvania were seropositive for S. neurona .

  16. Prevalence of antibodies against Neospora spp. and Sarcocystis neurona in donkeys from northeastern Brazil.

    Science.gov (United States)

    Gennari, Solange Maria; Pena, Hilda Fátima de Jesus; Lindsay, David Scott; Lopes, Marcos Gomes; Soares, Herbert Sousa; Cabral, Aline Diniz; Vitaliano, Sérgio Netto; Amaku, Marcos

    2016-01-01

    Sarcocystis neurona and Neospora hughesi are coccidian protozoa that can cause neurological illness in horses in America. In this study we report seroprevalence of Neospora spp. andS. neurona in sera of 333 donkeys from the northeastern region of Brazil. Antibodies to Neospora spp. were detected in 2% (7 donkeys) of 333 sera tested by the indirect fluorescent antibody test (IFAT) with a cut-off dilution of 1:40. Antibodies to S. neurona were found in 3% (10 donkeys) of the samples tested by IFAT (cut-off ≥50) and 21% (69 donkeys) by the direct agglutination test (SAT ≥50). The SAT and IFAT results for S. neurona showed a poor concordance (value of Kappa=0.051). This is the first report of Neospora spp. antibodies in Brazilian donkeys and the first detection of antibodies against S. neurona in this animal species.

  17. Serological investigation of transplacental infection with Neospora hughesi and Sarcocystis neurona in broodmares.

    Science.gov (United States)

    Pusterla, Nicola; Mackie, Sarah; Packham, Andrea; Conrad, Patricia A

    2014-12-01

    The aim of the present study was to investigate the likelihood of transplacental transmission of Neospora hughesi and Sarcocystis neurona in foals, born from seropositive mares. Three broodmares with persistent N. hughesi infection gave birth to eight healthy foals over a period of 7 years. These foals were seropositive to N. hughesi prior to colostrum ingestion, with titers ranging between 640 and 20,480, measured by indirect fluorescence antibody test (IFAT). Of 174 foals born at another farm to mares with a high seroprevalence to S. neurona, only one (with a pre-colostrum antibody titer of 80) tested seropositive. Transplacental transmission of N. hughesi seems to occur from latently infected mares to their foals, while this route of transmission does not seem to occur commonly for S. neurona. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Antibody index and specific antibody quotient in horses after intragastric administration of Sarcocystis neurona sporocysts.

    Science.gov (United States)

    Heskett, Katherine A; Mackay, Robert J

    2008-03-01

    To investigate the use of a specific antibody index (AI) that relates Sarcocystis neurona-specific IgG quotient (Q(SN)) to total IgG quotient (Q(IgG)) for the detection of the anti-S neurona antibody fraction of CNS origin in CSF samples obtained from horses after intragastric administration of S neurona sporocysts. 18 adult horses. 14 horses underwent intragastric inoculation (day 0) with S neurona sporocysts, and 4 horses remained unchallenged; blood and CSF samples were collected on days - 1 and 84. For purposes of another study, some challenged horses received intermittent administration of ponazuril (20 mg/kg, PO). Sarcocystis neurona-specific IgG concentrations in CSF (SN(CSF)) and plasma (SN(plasma)) were measured via a direct ELISA involving merozoite lysate antigen and reported as ELISA units (EUs; arbitrary units based on a nominal titer for undiluted immune plasma of 100,000 EUs/mL). Total IgG concentrations in CSF (IgG(CSF)) and plasma (IgG(plasma)) were quantified via a sandwich ELISA and a radial immunodiffusion assay, respectively; Q(SN), Q(IgG), and AI were calculated. Following sporocyst challenge, mean +/- SEM SN(CSF) and SN(plasma) increased significantly (from 8.8 +/- 1.0 EUs/mL to 270.0 +/- 112.7 EUs/mL and from 1,737 +/- 245 EUs/mL to 43,169 +/- 13,770 EUs/mL, respectively). Challenge did not affect total IgG concentration, Q(SN), Q(IgG), or AI. S neurona-specific IgG detected in CSF samples from sporocyst-challenged horses appeared to be extraneural in origin; thus, this experimental challenge may not reliably result in CNS infection. Calculation of a specific AI may have application to the diagnosis of S neurona-associated myeloencephalitis in horses.

  19. Parasitemia in an immunocompetent horse experimentally challenged with Sarcocystis neurona sporocysts.

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    Rossano, M G; Schott, H C; Murphy, A J; Kaneene, J B; Sellon, D C; Hines, M T; Hochstatter, T; Bell, J A; Mansfield, L S

    2005-01-04

    Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in Americans. Most cases are attributed to infection of the central nervous system with Sarcocystis neurona. Parasitemia has not been demonstrated in immunocompetent horses, but has been documented in one immunocompromised foal. The objective of this study was to isolate viable S. neurona from the blood of immunocompetent horses. Horses used in this study received orally administered S. neurona sporocysts (strain SN 37-R) daily for 112 days at the following doses: 100/day for 28 days, followed by 500/day for 28 days, followed by 1000/day for 56 days. On day 98 of the study, six yearling colts were selected for attempted culture of S. neurona from blood, two testing positive, two testing suspect and two testing negative for antibodies against S. neurona on day 84 of the study. Two 10 ml tubes with EDTA were filled from each horse by jugular venipuncture and the plasma fraction rich in mononuclear cells was pipetted onto confluent equine dermal cell cultures. The cultures were monitored weekly for parasite growth for 12 weeks. Merozoites grown from cultures were harvested and tested using S. neurona-specific PCR with RFLP to confirm species identity. PCR products were sequenced and compared to known strains of S. neurona. After 38 days of in vitro incubation, one cell culture from a horse testing positive for antibodies against S. neurona was positive for parasite growth while the five remaining cultures remained negative for parasite growth for all 12 weeks. The Sarcocystis isolate recovered from cell culture was confirmed to be S. neurona by PCR with RFLP. Gene sequence analysis revealed that the isolate was identical to the challenge strain SN-37R and differed from two known strains UCD1 and MIH1. To our knowledge this is the first report of parasitemia with S. neurona in an immunocompetent horse.

  20. Use of proteinase K in the excystation of Sarcocystis cruzi sporocysts for in vitro culture and DNA extraction.

    Science.gov (United States)

    Ndiritu, W; Cawthorn, R J; Kibenge, F S

    1994-03-01

    Proteinase K was used for the cleaning of Sarcocystis cruzi (Apicomplexa) sporocysts prior to excystation. Bovine pulmonary endothelial cell cultures inoculated with the excysted sporozoites remained free of bacterial contamination for the duration of the experiment and had high yields of merozoites. The excysted sporozoites also yielded genomic DNA that could be labelled efficiently with 32P dATP by the random priming method.

  1. Ongoing outbreak of an acute muscular Sarcocystis-like illness among travellers returning from Tioman Island, Malaysia, 2011-2012.

    Science.gov (United States)

    Esposito, D H; Freedman, D O; Neumayr, A; Parola, P

    2012-11-08

    As of 4 November, 2012, 100 patients with an acute muscular Sarcocystis-like illness associated with travel to Tioman Island, Malaysia, have been identified. Thirty-five travelled there mostly during July and August 2011 and 65 mostly during July and August 2012, suggesting an ongoing outbreak. Epidemiological investigations are ongoing. Public health agencies and practicing clinicians should be aware of this rarely-reported disease in humans and consider it as differential diagnosis in travellers returning from Tioman Island.

  2. A genetically diverse but distinct North American population of Sarcocystis neurona includes an overrepresented clone described by 12 microsatellite alleles.

    Science.gov (United States)

    Asmundsson, Ingrid M; Dubey, J P; Rosenthal, Benjamin M

    2006-09-01

    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.

  3. A report of intestinal sarcocystosis in the bullsnake (Pituophis melanoleucus sayi) and a re-evaluation of Sarcocystis sp. from snakes of the genus Pituophis.

    Science.gov (United States)

    Daszak, P; Cunningham, A

    1995-07-01

    We report a severe enteric infection of Sarcocystis sp. from a wild-caught bullsnake (Pituophis melanoleucus sayi). The animal was collected in October 1988 by a commercial dealer, imported into the United Kingdom during November 1988 and purchased by the London Zoo, in December 1988. The animal was not fed after capture and was anorexic from the time of purchase to the time of death in January 1989. On necropsy, the animal was emaciated and the mucosa of the proximal intestine was markedly thickened. The lamina propria was packed with oocysts, and enterocytes were parasitized by an organism which closely resembled Sarcocystis roudabushi and Sarcocystis idahoensis, two bisporocystid coccidia described previously from Pituophis melanoleucus. We propose that Sarcocystis idahoensis and Sarcocystis roudabushi are synonymous since both occur in the same host species, both invade the intestinal lamina propria and entreocytes, and sporocyst measurement ranges of both species overlap. This is the first report of death believed to be due to sarcocytosis in a naturally-infected definitive host.

  4. Prevalence of antibodies to Sarcocystis neurona and Neospora hughesi in horses from Mexico.

    Science.gov (United States)

    Yeargan, Michelle R; Alvarado-Esquivel, Cosme; Dubey, Jitender P; Howe, Daniel K

    2013-01-01

    Equine protozoal myeloencephalitis (EPM) is a debilitating disease of horses caused by Sarcocystis neurona and Neospora hughesi. Sera from 495 horses in Durango State, Mexico were tested for anti-protozoal antibodies using enzyme-linked immunosorbent assays (ELISAs) based on major surface antigens of these two parasites. Antibodies to S. neurona were detected in 240 (48.5%) of the 495 horse sera tested with the rSnSAG2/4/3 trivalent ELISA. Multivariate analysis showed that exposure to S. neurona was associated with age, feeding grains and crops, and small herd size. Antibodies to N. hughesi were found in 15 (3.0%) of the 495 horse sera tested with the rNhSAG1 ELISA and confirmed by Western blot of N. hughesi tachyzoite antigen. This is the first report of S. neurona and N. hughesi exposure in horses in Mexico, and it affirms that EPM should be in the differential diagnosis for horses exhibiting signs of neurologic disease in this country. © M.R. Yeargan et al., published by EDP Sciences, 2013.

  5. Prevalence and risk factors associated with Sarcocystis neurona infections in opossums (Didelphis virginiana) from central California.

    Science.gov (United States)

    Rejmanek, Daniel; Vanwormer, Elizabeth; Miller, Melissa A; Mazet, Jonna A K; Nichelason, Amy E; Melli, Ann C; Packham, Andrea E; Jessup, David A; Conrad, Patricia A

    2009-12-03

    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).

  6. In vitro efficacy of nitro- and halogeno-thiazolide/thiadiazolide derivatives against Sarcocystis neurona.

    Science.gov (United States)

    Gargala, G; Le Goff, L; Ballet, J J; Favennec, L; Stachulski, A V; Rossignol, J F

    2009-06-10

    Sarcocystis neurona is an obligate intracellular parasite that causes equine protozoal myeloencephalitis (EPM). The aim of this work was to document inhibitory activities of nitazoxanide (NTZ, [2-acetolyloxy-N-(5-nitro 2-thiazolyl) benzamide]) and new thiazolides/thiadiazolides on S. neurona in vitro development, and investigate their structure-activity relationships. S. neurona was grown in bovine turbinate cell cultures. At concentrations varying from 1.0 to 5.0mg/L, nitazoxanide and 21 of 32 second generation thiazolide/thiadiazolide agents exerted a > or =95% maximum inhibition on S. neurona development. Most active agents were either NO(2) or halogen substituted in position 5 of their thiazole moiety. In contrast, other 5-substitutions such as hydrogen, methyl, SO(2)CH(3), and CH(3) negatively impacted activity. Compared with derivatives with an acetylated benzene moiety, deacetylated compounds which most probably represent primary metabolites exhibited similar inhibitory activities. Present data provide the first evidence of in vitro inhibitory activities of nitazoxanide and new thiazolides/thiadiazolides on S. neurona development. Active halogeno-thiazolide/thiadiazolides may provide a valuable nitro-free alternative to nitazoxanide for EPM treatment depending on further evaluation of their in vivo activities.

  7. The identification of a sequence related to apicomplexan enolase from Sarcocystis neurona.

    Science.gov (United States)

    Wilson, A P; Thelen, J J; Lakritz, J; Brown, C R; Marsh, A E

    2004-11-01

    Equine protozoal myeloencephalitis (EPM) is a neurological disease caused by Sarcocystis neurona, an apicomplexan parasite. S. neurona is also associated with EPM-like diseases in marine and small mammals. The mechanisms of transmission and ability to infect a wide host range remain obscure; therefore, characterization of essential proteins may provide evolutionary information allowing the development of novel chemotherapeutics that target non-mammalian biochemical pathways. In the current study, two-dimensional electrophoresis and matrix-assisted laser desorption ionization-time of flight (MALDI-ToF) mass spectrometry were combined to characterize and identify an enolase protein from S. neurona based on peptide homology to the Toxoplasma gondii protein. Enolase is thought to be a vestigial, non-photosynthetic protein resulting from an evolutionary endosymbiosis event of an apicomplexan ancestor with green algae. Enolase has also been suggested to play a role in parasite stage conversion for T. gondii. Characterization of this protein in S. neurona and comparison to other protozoans indicate a biochemical similarity of S. neurona enolase to other tissue-cyst forming coccidians that cause encephalitis.

  8. Seroepidemiology of Sarcocystis neurona and Neospora hughesi infections in domestic donkeys (Equus asinus in Durango, Mexico

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    Alvarado-Esquivel Cosme

    2017-01-01

    Full Text Available There is currently no information regarding Sarcocystis neurona and Neospora hughesi infections in donkeys in Mexico. Here, we determined the presence of antibodies against S. neurona and N. hughesi in donkeys in the northern Mexican state of Durango. Serum samples of 239 domestic donkeys (Equus asinus were assayed for S. neurona and N. hughesi antibodies using home-made enzyme-linked immunoassays; six (2.5% of the 239 donkeys tested seropositive for S. neurona. The seroprevalence of S. neurona infection was comparable among donkeys regardless of their origin, health status, or sex. Multivariate analysis showed that seropositivity to S. neurona was associated with increased age (OR = 2.95; 95% CI: 1.11–7.82; p = 0.02. Antibodies to N. hughesi were found in two (0.8% of the 239 donkeys. Both exposed donkeys were healthy, 3- and 6-year-old females. This is the first evidence of S. neurona and N. hughesi infections in donkeys in Mexico.

  9. Molecular characterization of Sarcocystis neurona strains from opossums (Didelphis virginiana) and intermediate hosts from Central California.

    Science.gov (United States)

    Rejmanek, Daniel; Miller, Melissa A; Grigg, Michael E; Crosbie, Paul R; Conrad, Patricia A

    2010-05-28

    Sarcocystis neurona is a significant cause of neurological disease in horses and other animals, including the threatened Southern sea otter (Enhydra lutris nereis). Opossums (Didelphis virginiana), the only known definitive hosts for S. neurona in North America, are an introduced species in California. S. neurona DNA isolated from sporocysts and/or infected tissues of 10 opossums, 6 horses, 1 cat, 23 Southern sea otters, and 1 harbor porpoise (Phocoena phocoena) with natural infections was analyzed based on 15 genetic markers, including the first internal transcribed spacer (ITS-1) region; the 25/396 marker; S. neurona surface antigen genes (snSAGs) 2, 3, and 4; and 10 different microsatellites. Based on phylogenetic analysis, most of the S. neurona strains segregated into three genetically distinct groups. Additionally, fifteen S. neurona samples from opossums and several intermediate hosts, including sea otters and horses, were found to be genetically identical across all 15 genetic markers, indicating that fatal encephalitis in Southern sea otters and equine protozoal myeloencephalitis (EPM) in horses is strongly linked to S. neurona sporocysts shed by opossums. (c) 2010 Elsevier B.V. All rights reserved.

  10. Has Sarcocystis neurona Dubey et al., 1991 (Sporozoa: Apicomplexa: Sarcocystidae) cospeciated with its intermediate hosts?

    Science.gov (United States)

    Elsheikha, Hany M

    2009-08-26

    The question of how Sarcocystis neurona is able to overcome species barrier and adapt to new hosts is central to the understanding of both the evolutionary origin of S. neurona and the prediction of its field host range. Therefore, it is worth reviewing current knowledge on S. neurona host specificity. The available host range data for S. neurona are discussed in relation to a subject of evolutionary importance-specialist or generalist and its implications to understand the strategies of host adaptation. Current evidences demonstrate that a wide range of hosts exists for S. neurona. This parasite tends to be highly specific for its definitive host but much less so for its intermediate host (I.H.). The unique specificity of S. neurona for its definitive host may be mediated by a probable long coevolutionary relationship of the parasite and carnivores in a restricted ecological niche 'New World'. This might be taken as evidence that carnivores are the 'original' host group for S. neurona. Rather, the capacity of S. neurona to exploit an unusually large number of I.H. species probably indicates that S. neurona maintains non-specificity to its I.H. as an adaptive response to insure the survival of the parasite in areas in which the 'preferred' host is not available. This review concludes with the view that adaptation of S. neurona to a new host is a complex interplay that involves a large number of determinants.

  11. Immune response to Sarcocystis neurona infection in naturally infected horses with equine protozoal myeloencephalitis.

    Science.gov (United States)

    Yang, Jibing; Ellison, Siobhan; Gogal, Robert; Norton, Heather; Lindsay, David S; Andrews, Frank; Ward, Daniel; Witonsky, Sharon

    2006-06-15

    Equine protozoal myeloencephalitis (EPM) is one of the most common neurologic diseases of horses in the United States. The primary etiologic agent is Sarcocystis neurona. Currently, there is limited knowledge regarding the protective or pathophysiologic immune response to S. neurona infection or the subsequent development of EPM. The objectives of this study were to determine whether S. neurona infected horses with clinical signs of EPM had altered or suppressed immune responses compared to neurologically normal horses and if blood sample storage would influence these findings. Twenty clinically normal horses and 22 horses with EPM, diagnosed by the presence of S. neurona specific antibodies in the serum and/or cerebrospinal (CSF) and clinical signs, were evaluated for differences in the immune cell subsets and function. Our results demonstrated that naturally infected horses had significantly (Pneurona in horses, as well as to determine the mechanism associated with suppressed in vitro proliferation responses. Finally, overnight storage of blood samples appears to alter T lymphocyte phenotypes and viability among leukocytes.

  12. Risk of transplacental transmission of Sarcocystis neurona and Neospora hughesi in California horses.

    Science.gov (United States)

    Duarte, Paulo C; Conrad, Patricia A; Barr, Bradd C; Wilson, W David; Ferraro, Gregory L; Packham, Andrea E; Carpenter, Tim E; Gardner, Ian A

    2004-12-01

    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 neurona and N. hughesi in mares increased with age. Mares 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.

  13. Horses experimentally infected with Sarcocystis neurona develop altered immune responses in vitro.

    Science.gov (United States)

    Witonsky, Sharon G; Ellison, Siobhan; Yang, Jibing; Gogal, Robert M; Lawler, Heather; Suzuki, Yasuhiro; Sriranganathan, Namalwar; Andrews, Frank; Ward, Daniel; Lindsay, David S

    2008-10-01

    Equine protozoal myeloencephalitis (EPM) due to Sarcocystis neurona infection is 1 of the most common neurologic diseases in horses in the United States. The mechanisms by which most horses resist disease, as well as the possible mechanisms by which the immune system may be suppressed in horses that develop EPM, are not known. Therefore, the objectives of this study were to determine whether horses experimentally infected with S. neurona developed suppressed immune responses. Thirteen horses that were negative for S. neurona antibodies in serum and cerebrospinal fluid (CSF) were randomly assigned to control (n = 5) or infected (n = 8) treatment groups. Neurologic exams and cerebrospinal fluid analyses were performed prior to, and following, S. neurona infection. Prior to, and at multiple time points following infection, immune parameters were determined. All 8 S. neurona-infected horses developed clinical signs consistent with EPM, and had S. neurona antibodies in the serum and CSF. Both infected and control horses had increased percentages (P < 0.05) of B cells at 28 days postinfection. Infected horses had significantly decreased (P < 0.05) proliferation responses as measured by thymidine incorporation to nonspecific mitogens phorbol myristate acetate (PMA) and ionomycin (I) as soon as 2 days postinfection.

  14. Seroepidemiology of Sarcocystis neurona and Neospora hughesi infections in domestic donkeys (Equus asinus) in Durango, Mexico.

    Science.gov (United States)

    Alvarado-Esquivel, Cosme; Howe, Daniel K; Yeargan, Michelle R; Alvarado-Esquivel, Domingo; Alfredo Zamarripa-Barboza, José; Dubey, Jitender P

    2017-01-01

    There is currently no information regarding Sarcocystis neurona and Neospora hughesi infections in donkeys in Mexico. Here, we determined the presence of antibodies against S. neurona and N. hughesi in donkeys in the northern Mexican state of Durango. Serum samples of 239 domestic donkeys (Equus asinus) were assayed for S. neurona and N. hughesi antibodies using home-made enzyme-linked immunoassays; six (2.5%) of the 239 donkeys tested seropositive for S. neurona. The seroprevalence of S. neurona infection was comparable among donkeys regardless of their origin, health status, or sex. Multivariate analysis showed that seropositivity to S. neurona was associated with increased age (OR = 2.95; 95% CI: 1.11-7.82; p = 0.02). Antibodies to N. hughesi were found in two (0.8%) of the 239 donkeys. Both exposed donkeys were healthy, 3- and 6-year-old females. This is the first evidence of S. neurona and N. hughesi infections in donkeys in Mexico. © C. Alvarado-Esquivel et al., published by EDP Sciences, 2017.

  15. California mussels (Mytilus californianus) as sentinels for marine contamination with Sarcocystis neurona.

    Science.gov (United States)

    Michaels, Lauren; Rejmanek, Daniel; Aguilar, Beatriz; Conrad, Patricia; Shapiro, Karen

    2016-05-01

    Sarcocystis neurona is a terrestrial parasite that can cause fatal encephalitis in the endangered Southern sea otter (Enhydra lutris nereis). To date, neither risk factors associated with marine contamination nor the route of S. neurona infection to marine mammals has been described. This study evaluated coastal S. neurona contamination using California mussels (Mytilus californianus) as sentinels for pathogen pollution. A field investigation was designed to test the hypotheses that (1) mussels can serve as sentinels for S. neurona contamination, and (2) S. neurona contamination in mussels would be highest during the rainy season and in mussels collected near freshwater. Initial validation of molecular assays through sporocyst spiking experiments revealed the ITS-1500 assay to be most sensitive for detection of S. neurona, consistently yielding parasite amplification at concentrations ⩾5 sporocysts/1 mL mussel haemolymph. Assays were then applied on 959 wild-caught mussels, with detection of S. neurona confirmed using sequence analysis in three mussels. Validated molecular assays for S. neurona detection in mussels provide a novel toolset for investigating marine contamination with this parasite, while confirmation of S. neurona in wild mussels suggests that uptake by invertebrates may serve as a route of transmission to susceptible marine animals.

  16. Detection of antibodies against Sarcocystis neurona, Neospora spp., and Toxoplasma gondii in horses from Costa Rica.

    Science.gov (United States)

    Dangoudoubiyam, S; Oliveira, J B; Víquez, C; Gómez-García, A; González, O; Romero, J J; Kwok, O C H; Dubey, J P; Howe, D K

    2011-06-01

    Serum samples from 315 horses from Costa Rica, Central America, were examined for the presence of antibodies against Sarcocystis neurona, Neospora spp., and Toxoplasma gondii by using the surface antigen (SAG) SnSAG2 enzyme-linked immunosorbent assay (ELISA), the NhSAG1 ELISA, and the modified agglutination test, respectively. Anti- S. neurona antibodies were found in 42.2% of the horses by using the SnSAG2 ELISA. Anti- Neospora spp. antibodies were found in only 3.5% of the horses by using the NhSAG1 ELISA, and only 1 of these horses was confirmed seropositive by Western blot. Antibodies to T. gondii were found in 34.0% of the horses tested, which is higher than in previous reports from North and South America. The finding of anti- S. neurona antibodies in horses from geographical areas where Didelphis marsupialis has wide distribution suggests that D. marsupialis is a potential definitive host for this parasite and a source of infection for these horses.

  17. Dual Sarcocystis neurona and Toxoplasma gondii infection in a northern sea otter from Washington state, USA

    Science.gov (United States)

    Lindsay, D.S.; Thomas, N.J.; Rosypal, A.C.; Dubey, J.P.

    2001-01-01

    Dual Sarcocystis neurona and Toxoplasma gondii infection was observed in a Northern sea otter from Washington, USA. The animal was found stranded, convulsed, and died shortly thereafter. Encephalitis caused by both S. neurona and T. gondii was demonstrated in histological sections of brain. Immunohistochemical examination of sections with S. neurona specific antisera demonstrated developmental stages that divided by endopolygeny and produced numerous merozoites. PCR of brain tissue from the sea otter using primer pairs JNB33/JNB54 resulted in amplification of a 1100 bp product. This PCR product was cut in to 884 and 216 bp products by Dra I but was not cut by Hinf I indicating that it was S. neurona [J. Parasitol. 85 (1999) 221]. No PCR product was detected in the brain of a sea otter which had no lesions of encephalitis. Examination of brain sections using T. gondii specific antisera demonstrated tachyzoites and tissue cysts of T. gondii. The lesions induced by T. gondii suggested that the sea otter was suffering from reactivated toxoplasmosis. T. gondii was isolated in mice inoculated with brain tissue. A cat that was fed infected mouse brain tissue excreted T. gondii oocysts which were infective for mice. This is apparently the first report of dual S. neurona and T. gondii in a marine mammal.

  18. Infection of immunodeficient horses with Sarcocystis neurona does not result in neurologic disease.

    Science.gov (United States)

    Sellon, Debra C; Knowles, Donald P; Greiner, Ellis C; Long, Maureen T; Hines, Melissa T; Hochstatter, Tressa; Tibary, Ahmed; Dame, John B

    2004-11-01

    Equine protozoal myeloencephalitis is a progressive neurologic disease of horses most commonly caused by infection with the apicomplexan parasite Sarcocystis neurona. Factors affecting neuroinvasion and neurovirulence have not been determined. We investigated the pathogenesis of infection with S. neurona in horses with severe combined immune deficiency (SCID). Two immunocompetent (IC) Arabian horses and two Arabian horses with SCID were infected orally with 5 x 10(5) sporocysts of S. neurona. Four IC horses and one SCID horse were infected intravenously (i.v.) with 5 x 10(8) merozoites of the WSU-1 isolate of S. neurona. Despite prolonged parasitemia and persistent infection of visceral tissues (skeletal muscle, cardiac muscle, lung, liver, and spleen) as demonstrated by PCR and culture, SCID horses did not develop neurologic signs after oral or i.v. infection. S. neurona was undetectable in the neuronal tissues of SCID horses by either PCR, immunohistochemistry, or culture. In contrast, although parasitemia was undetectable in orally infected IC horses and of only short duration in i.v. infected IC horses, four of six IC horses developed neurologic signs. S. neurona was detectable by PCR and/or culture of neural tissue but not visceral tissue of IC horses with neurologic disease. Infected SCID horses are unable to clear S. neurona from visceral tissues, but the infection does not result in neurologic signs; in contrast, IC horses rapidly control parasitemia and infection of visceral tissues but frequently experience neuroinvasion and exhibit clinical signs of neurologic disease.

  19. Effect of intermittent oral administration of ponazuril on experimental Sarcocystis neurona infection of horses.

    Science.gov (United States)

    Mackay, Robert J; Tanhauser, Susan T; Gillis, Karen D; Mayhew, Ian G; Kennedy, Tom J

    2008-03-01

    To evaluate the effect of intermittent oral administration of ponazuril on immunoconversion against Sarcocystis neurona in horses inoculated intragastrically with S neurona sporocysts. 20 healthy horses that were seronegative for S neurona-specific IgG. 5 control horses were neither inoculated with sporocysts nor treated. Other horses (5 horses/group) each received 612,500 S neurona sporocysts via nasogastric tube (day 0) and were not treated or were administered ponazuril (20 mg/kg, PO) every 7 days (beginning on day 5) or every 14 days (beginning on day 12) for 12 weeks. Blood and CSF samples were collected on day - 1 and then every 14 days after challenge for western blot assessment of immunoconversion. Clinical signs of equine protozoal myeloencephalitis (EPM) were monitored, and tissues were examined histologically after euthanasia. Sera from all challenged horses yielded positive western blot results within 56 days. Immunoconversion in CSF was detected in only 2 of 5 horses that were treated weekly; all other challenged horses immunoconverted within 84 days. Weekly administration of ponazuril significantly reduced the antibody response against the S neurona 17-kd antigen in CSF. Neurologic signs consistent with EPM did not develop in any group; likewise, histologic examination of CNS tissue did not reveal protozoa or consistent degenerative or inflammatory changes. Administration of ponazuril every 7 days, but not every 14 days, significantly decreased intrathecal anti-S neurona antibody responses in horses inoculated with S neurona sporocysts. Protocols involving intermittent administration of ponazuril may have application in prevention of EPM.

  20. Sarcocystis schneideri n. sp. (Sarcocystidae) infecting the barber skink Eumeces schneideri schneideri (Scincidae) Daudin, 1802. A light and ultrastructural study.

    Science.gov (United States)

    Bashtar, Abdel-Rahman; Al Aal, Zain Abd; Maarouf, Wael; Morsy, Kareem; Al Quraishy, Saleh

    2014-06-01

    The current study provides the first record of infection with Sarcocystis species in the barber skink Eumeces schneideri schneideri (Scincidae) captured from the north region of Egypt around the cities of El-Hamam and Al-Dabaa, Mersa Matruh Governorate, Egypt. Morphology of the parasite cysts was described using light and transmission electron microscopy. Five out of 80 (6.25%) of the examined skinks were found to be infected. The infection was recorded firstly by light microscopy as spindle-shaped cysts embedded in the muscle tissue. The cysts were microscopic and measured 250-900 μm in length × 50-100 μm in width (mean, 575 × 75 μm). The validity of this species was confirmed by means of ultrastructural characteristics of the primary cyst wall (0.28 μm thick) which revealed the presence of irregularly shaped crowded and osmiophilic knob-like projections underlined by a thin layer of ground substance measuring 0.15-0.17 μm (mean, 0.16 μm). 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 3-5 μm in length and 1.5-2.5 in width with centrally or posteriorly located nuclei. The morphological and morphometric data obtained during study were compared with those recorded previously from organisms within the Scincidae family. It was observed that this parasite possessed some distinguishing characteristics from the comparable species, which should be considered as a new species of the Sarcocystis genus, and the proposed name was Sarcocystis schneideri n. sp. with new host and locality records in Egypt.

  1. Acute, fatal Sarcocystis calchasi-associated hepatitis in Roller pigeons (Columba livia f. dom.) at Philadelphia Zoo.

    Science.gov (United States)

    Trupkiewicz, J G; Calero-Bernal, R; Verma, S K; Mowery, J; Davison, S; Habecker, P; Georoff, T A; Ialeggio, D M; Dubey, J P

    2016-01-30

    Four Roller pigeons (Columba livia f. dom.) at the Philadelphia Zoo died suddenly. Necropsy examination revealed macroscopic hepatitis. Microscopically, the predominant lesions were in liver, characterized with necrosis and mixed cell inflammatory response. Sarcocystis calchasi-like schizonts and free merozoites were identified in liver. Transmission electron microscopy confirmed that schizonts were in hepatocytes. A few schizonts were in spleen. PCR using S. calchasi-specific primers confirmed the diagnosis. Neither lesions nor protozoa were found in brain and muscles. This is the first report of acute visceral S. calchasi-associated sarcocystosis in naturally infected avian hosts.

  2. Cross-sectional survey in pig breeding farms in Hesse, Germany: seroprevalence and risk factors of infections with Toxoplasma gondii, Sarcocystis spp. and Neospora caninum in sows

    DEFF Research Database (Denmark)

    Damriyasa, I.M.; Bauer, C.; Edelhofer, R.

    2004-01-01

    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 plas...

  3. Sarcocystis pantherophis, n. sp. from eastern rat snakes (Pantherophis alleghaniensis) definitive hosts and interferongamma gene knockout mice as experimental intermediate hosts

    Science.gov (United States)

    Here we report a new species, Sarcocystis pantherophisi with the Eastern rat snake (Pantherophis alleghaniensis) as natural definitive host and the interferon gamma gene knockout (KO) mouse as the experimental intermediate host. Sporocysts (n=15) from intestinal contents of the snake were 17.3 x 10....

  4. In the United States, negligible rates of zoonotic sarcocystosis occur in feral swine that, by contrast, frequently harbor infections with Sarcocystis meischeriana, a related parasite contracted from dogs

    Science.gov (United States)

    Transmission of pathogens between domestic and wild life animals plays important role in epidemiology. Feral pig populations are increasing and expanding in the USA, and may constitute a risk to non-biosecure domestic pig facilities by serving as reservoirs for pathogens. We surveyed, for Sarcocysti...

  5. Occurrence of Sarcocystis spp. in opossums (Didelphis aurita and Didelphis albiventris in regions of the State of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Renata Assis Casagrande

    2009-04-01

    Full Text Available O objetivo deste estudo foi de determinar a ocorrência de Sarcocystis spp. em D. albiventris e D. aurita em três regiões do Estado de São Paulo. Para tal, utilizou-se noventa e oito Didelphis mortos, sendo 66 D. aurita e 32 D. albiventris, e também 28 D. aurita e cinco D. albiventris vivos. O método de centrífugo-flutuação em solução de sacarose foi empregado para isolamento dos oocistos/esporocistos de Sarcocystis spp. do intestino delgado e das fezes. Encontrou-se Sarcocystis spp. no intestino delgado de 9,1% dos D. aurita (6/66, sendo que quatro destes também houve positividade nas fezes. Não houve diferença estatística significativa entre machos e fêmeas positivos (P= 0,522, e entre os positivos de diferentes origens do Estado de São Paulo (P= 0,627, quanto a ocorrência de Sarcocystis spp. Entretanto, houve diferença estatística significativa entre animais de vida livre e de cativeiro (P = 0.009, sendo que somente os de vida livre foram positivos. Entre adultos e filhotes positivos também houve diferença (P= 0,004, sendo os adultos mais parasitados que os filhotes. Das amostras provenientes dos 28 D. aurita vivos, encontrou-se Sarcocystis spp. em 7.1% (2/28 deles. Dos 32 D. albiventris, todos foram negativos para Sarcocystis spp. nas amostras de intestino delgado e fezes. Os cincos D. albiventris vivos também foram negativos. Sendo assim, pode-se observar que a ocorrência de Sarcocystis spp. em D. aurita e D. albiventris nestas três regiões do Estado de São Paulo é baixa para estas condições analisadas.

  6. Genome-Wide Identification and Evolutionary Analysis of Sarcocystis neurona Protein Kinases

    Directory of Open Access Journals (Sweden)

    Edwin K. Murungi

    2017-03-01

    Full Text Available The apicomplexan parasite Sarcocystis neurona causes equine protozoal myeloencephalitis (EPM, a degenerative neurological disease of horses. Due to its host range expansion, S. neurona is an emerging threat that requires close monitoring. In apicomplexans, protein kinases (PKs have been implicated in a myriad of critical functions, such as host cell invasion, cell cycle progression and host immune response evasion. Here, we used various bioinformatics methods to define the kinome of S. neurona and phylogenetic relatedness of its PKs to other apicomplexans. We identified 97 putative PKs clustering within the various eukaryotic kinase groups. Although containing the universally-conserved PKA (AGC group, S. neurona kinome was devoid of PKB and PKC. Moreover, the kinome contains the six-conserved apicomplexan CDPKs (CAMK group. Several OPK atypical kinases, including ROPKs 19A, 27, 30, 33, 35 and 37 were identified. Notably, S. neurona is devoid of the virulence-associated ROPKs 5, 6, 18 and 38, as well as the Alpha and RIO kinases. Two out of the three S. neurona CK1 enzymes had high sequence similarities to Toxoplasma gondii TgCK1-α and TgCK1-β and the Plasmodium PfCK1. Further experimental studies on the S. neurona putative PKs identified in this study are required to validate the functional roles of the PKs and to understand their involvement in mechanisms that regulate various cellular processes and host-parasite interactions. Given the essentiality of apicomplexan PKs in the survival of apicomplexans, the current study offers a platform for future development of novel therapeutics for EPM, for instance via application of PK inhibitors to block parasite invasion and development in their host.

  7. Investigation of SnSPR1, a novel and abundant surface protein of Sarcocystis neurona merozoites.

    Science.gov (United States)

    Zhang, Deqing; Howe, Daniel K

    2008-04-15

    An expressed sequence tag (EST) sequencing project has produced over 15,000 partial cDNA sequences from the equine pathogen Sarcocystis neurona. While many of the sequences are clear homologues of previously characterized genes, a significant number of the S. neurona ESTs do not exhibit similarity to anything in the extensive sequence databases that have been generated. In an effort to characterize parasite proteins that are novel to S. neurona, a seemingly unique gene was selected for further investigation based on its abundant representation in the collection of ESTs and the predicted presence of a signal peptide and glycolipid anchor addition on the encoded protein. The gene was expressed in E. coli, and monospecific polyclonal antiserum against the recombinant protein was produced by immunization of a rabbit. Characterization of the native protein in S. neurona merozoites and schizonts revealed that it is a low molecular weight surface protein that is expressed throughout intracellular development of the parasite. The protein was designated Surface Protein 1 (SPR1) to reflect its display on the outer surface of merozoites and to distinguish it from the ubiquitous SAG/SRS surface antigens of the heteroxenous Coccidia. Interestingly, infection assays in the presence of the polyclonal antiserum suggested that SnSPR1 plays some role in attachment and/or invasion of host cells by S. neurona merozoites. The work described herein represents a general template for selecting and characterizing the various unidentified gene sequences that are plentiful in the EST databases for S. neurona and other apicomplexans. Furthermore, this study illustrates the value of investigating these novel sequences since it can offer new candidates for diagnostic or vaccine development while also providing greater insight into the biology of these parasites.

  8. Genome-Wide Identification and Evolutionary Analysis of Sarcocystis neurona Protein Kinases.

    Science.gov (United States)

    Murungi, Edwin K; Kariithi, Henry M

    2017-03-21

    The apicomplexan parasite Sarcocystis neurona causes equine protozoal myeloencephalitis (EPM), a degenerative neurological disease of horses. Due to its host range expansion, S. neurona is an emerging threat that requires close monitoring. In apicomplexans, protein kinases (PKs) have been implicated in a myriad of critical functions, such as host cell invasion, cell cycle progression and host immune response evasion. Here, we used various bioinformatics methods to define the kinome of S. neurona and phylogenetic relatedness of its PKs to other apicomplexans. We identified 97 putative PKs clustering within the various eukaryotic kinase groups. Although containing the universally-conserved PKA (AGC group), S. neurona kinome was devoid of PKB and PKC. Moreover, the kinome contains the six-conserved apicomplexan CDPKs (CAMK group). Several OPK atypical kinases, including ROPKs 19A, 27, 30, 33, 35 and 37 were identified. Notably, S. neurona is devoid of the virulence-associated ROPKs 5, 6, 18 and 38, as well as the Alpha and RIO kinases. Two out of the three S. neurona CK1 enzymes had high sequence similarities to Toxoplasma gondii TgCK1-α and TgCK1-β and the Plasmodium PfCK1. Further experimental studies on the S. neurona putative PKs identified in this study are required to validate the functional roles of the PKs and to understand their involvement in mechanisms that regulate various cellular processes and host-parasite interactions. Given the essentiality of apicomplexan PKs in the survival of apicomplexans, the current study offers a platform for future development of novel therapeutics for EPM, for instance via application of PK inhibitors to block parasite invasion and development in their host.

  9. Identification of a dithiol-dependent nucleoside triphosphate hydrolase in Sarcocystis neurona.

    Science.gov (United States)

    Zhang, Deqing; Gaji, Rajshekhar Y; Howe, Daniel K

    2006-09-01

    A putative nucleoside triphosphate hydrolase (NTPase) gene was identified in a database of expressed sequence tags (ESTs) from the apicomplexan parasite Sarcocystis neurona. Analysis of culture-derived S. neurona merozoites demonstrated a dithiol-dependent NTPase activity, consistent with the presence of a homologue to the TgNTPases of Toxoplasma gondii. A complete cDNA was obtained for the S. neurona gene and the predicted amino acid sequence shared 38% identity with the two TgNTPase isoforms from T. gondii. Based on the obvious homology, the S. neurona protein was designated SnNTP1. The SnNTP1 cDNA encodes a polypeptide of 714 amino acids with a predicted 22-residue signal peptide and an estimated mature molecular mass of 70kDa. Southern blot analysis of the SnNTP1 locus revealed that the gene exists as a single copy in the S. neurona genome, unlike the multiple gene copies that have been observed in T. gondii and Neospora caninum. Analyses of the SnNTP1 protein demonstrated that it is soluble and secreted into the culture medium by extracellular merozoites. Surprisingly, indirect immunofluorescence analysis of intracellular S. neurona revealed apical localisation of SnNTP1 and temporal expression characteristics that are comparable with the microneme protein SnMIC10. The absence of SnNTP1 during much of endopolygeny implies that this protein does not serve a function during intracellular growth and development of S. neurona schizonts. Instead, SnNTP1 may play a role in events that occur during or proximal to merozoite egress from and/or invasion into cells.

  10. Risk of postnatal exposure to Sarcocystis neurona and Neospora hughesi in horses.

    Science.gov (United States)

    Duarte, Paulo C; Conrad, Patricia A; Wilson, W David; Ferraro, Gregory L; Packham, Andrea E; Bowers-Lepore, Jeanne; Carpenter, Tim E; Gardner, Ian A

    2004-08-01

    To estimate risk of exposure and age at first exposure to Sarcocystis neurona and Neospora hughesi and time to maternal antibody decay in foals. 484 Thoroughbred and Warmblood foals from 4 farms in California. Serum was collected before and after colostrum ingestion and at 3-month intervals thereafter. Samples were tested by use of the indirect fluorescent antibody test; cutoff titers were > or = 40 and > or = 160 for S neurona and N hughesi, respectively. Risk of exposure to S neurona and N hughesi during the study were 8.2% and 3.1%, respectively. Annual rate of exposure was 3.1% for S neurona and 1.7% for N hughesi. There was a significant difference in the risk of exposure to S neurona among farms but not in the risk of exposure to N hughesi. Median age at first exposure was 1.2 years for S neurona and 0.8 years for N hughesi. Highest prevalence of antibodies against S neurona and N hughesi was 6% and 2.1 %, respectively, at a mean age of 1.7 and 1.4 years, respectively. Median time to maternal antibody decay was 96 days for S neurona and 91 days for N hughesi. There were no clinical cases of equine protozoal myeloenchaphlitis (EPM). Exposure to S neurona and N hughesi was low in foals between birth and 2.5 years of age. Maternally acquired antibodies may cause false-positive results for 3 or 4 months after birth, and EPM was a rare clinical disease in horses < or = 2.5 years of age.

  11. Diversity of Sarcocystis spp shed by opossums in Brazil inferred with phylogenetic analysis of DNA coding ITS1, cytochrome B, and surface antigens.

    Science.gov (United States)

    Valadas, Samantha Y O B; da Silva, Juliana I G; Lopes, Estela Gallucci; Keid, Lara B; Zwarg, Ticiana; de Oliveira, Alice S; Sanches, Thaís C; Joppert, Adriana M; Pena, Hilda F J; Oliveira, Tricia M F S; Ferreira, Helena L; Soares, Rodrigo M

    2016-05-01

    Although few species of Sarcocystis are known to use marsupials of the genus Didelphis as definitive host, an extensive diversity of alleles of surface antigen genes (sag2, sag3, and sag4) has been described in samples of didelphid opossums in Brazil. In this work, we studied 25 samples of Sarcocystis derived from gastrointestinal tract of opossums of the genus Didelphis by accessing the variability of sag2, sag3, sag4, gene encoding cytochrome b (cytB) and first internal transcribed spacer (ITS1). Reference samples of Sarcocystis neurona (SN138) and Sarcocystis falcatula (SF1) maintained in cell culture were also analyzed. We found four allele variants of cytB, seven allele variants of ITS1, 10 allele variants of sag2, 13 allele variants of sag3, and 6 allele variants of sag4. None of the sporocyst-derived sequences obtained from Brazilian opossums revealed 100% identity to SN138 at cytB gene, nor to SN138 or SF1 at ITS1 locus. In addition, none of the sag alleles were found identical to either SF1 or SN138 homologous sequences, and a high number of new sag allele types were found other than those previously described in Brazil. Out of ten sag2 alleles, four are novel, while eight out of 13 sag3 alleles are novel and one out of six sag4 alleles is novel. Further studies are needed to clarify if such a vast repertoire of allele variants of Sarcocystis is the consequence of re-assortments driven by sexual exchange, in order to form individuals with highly diverse characteristics, such as pathogenicity, host spectrum, among others or if it only represents allele variants of different species with different biological traits.

  12. Effect of daily administration of pyrantel tartrate in preventing infection in horses experimentally challenged with Sarcocystis neurona.

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    Rossano, Mary G; Schott, Harold C; Kaneene, John B; Murphy, Alice J; Kruttlin, Elizabeth A; Hines, Melissa T; Sellon, Debra C; Patterson, Jon S; Elsheikha, Hany M; Dubey, J P; Mansfield, Linda S

    2005-05-01

    To determine whether daily administration of pyrantel tartrate can prevent infection in horses experimentally challenged with Sarcocystis neurona. 24 mixed-breed specific-pathogen-free weanling horses, 10 adult horses, 1 opossum, and 6 mice. Sarcocystis neurona-naïve weanling horses were randomly allocated to 2 groups. Group A received pyrantel tartrate at the labeled dose, and group B received a nonmedicated pellet. Both groups were orally inoculated with 100 sporocysts/d for 28 days, 500 sporocysts/d for 28 days, and 1000 sporocysts/d for 56 days. Blood samples were collected weekly, and CSF was collected monthly. Ten seronegative adult horses were monitored as untreated, uninfected control animals. All serum and CSF samples were tested by use of western blot tests to detect antibodies against S. neurona. At the end of the study, the number of seropositive and CSF-positive horses in groups A and B were compared by use of the Fisher exact test. Time to seroconversion on the basis of treatment groups and sex of horses was compared in 2 univariable Cox proportional hazards models. After 134 days of sporocyst inoculation, no significant differences were found between groups A and B for results of western blot tests of serum or CSF There were no significant differences in number of days to seroconversion on the basis of treatment groups or sex of horses. The control horses remained seronegative. Daily administration of pyrantel tartrate at the current labeled dose does not prevent S. neurona infection in horses.

  13. The gamma interferon knockout mouse model for sarcocystis neurona: comparison of infectivity of sporocysts and merozoites and routes of inoculation.

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    Dubey, J P; Lindsay, D S; Kwok, O C; Shen, S K

    2001-10-01

    The dose-related infectivity of Sarcocystis neurona sporocysts and merozoites of 2 recent isolates of S. neurona was compared in gamma interferon knockout (KO) mice. Tenfold dilutions of sporocysts or merozoites were bioassayed in mice, cell culture, or both. All 8 mice, fed 1,000 sporocysts, developed neurological signs with demonstrable S. neurona in their tissues. Of 24 mice fed low numbers of sporocysts (100, 10, 1), 18 became ill by 4 wk postinoculation, and S. neurona was demonstrated in their brains; antibodies (S. neurona agglutination test) to S. neurona and S. neurona parasites were not found in tissues of the 6 mice that were fed sporocysts and survived for >39 days. One thousand culture-derived merozoites of these 2 isolates were pathogenic to all 8 mice inoculated subcutaneously (s.c.). Of the 24 mice inoculated s.c. with merozoites numbering 100, 10, or 1, only 3 mice had demonstrable S. neurona infection; antibodies to S. neurona were not found in the 21 mice that had no demonstrable organisms. As few as 10 merozoites were infective for cell cultures. These results demonstrate that at least 1,000 merozoites are needed to cause disease in KO mice. Sarcocystis neurona sporocysts were infective to mice by the s.c. route.

  14. Limited genetic diversity among Sarcocystis neurona strains infecting southern sea otters precludes distinction between marine and terrestrial isolates.

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    Wendte, J M; Miller, M A; Nandra, A K; Peat, S M; Crosbie, P R; Conrad, P A; Grigg, M E

    2010-04-19

    Sarcocystis neurona is an apicomplexan parasite identified as a cause of fatal neurological disease in the threatened southern sea otter (Enhydra lutris nereis). In an effort to characterize virulent S. neurona strains circulating in the marine ecosystem, this study developed a range of markers relevant for molecular genotyping. Highly conserved sequences within the 18S ribosomal gene array, the plastid-encoded RNA polymerase (RPOb) and the cytochrome c oxidase subunit 1 mitochondrial gene (CO1) were assessed for their ability to distinguish isolates at the genus and species level. For within-species comparisons, five surface antigens (SnSAG1-SnSAG5) and one high resolution microsatellite marker (Sn9) were developed as genotyping markers to evaluate intra-strain diversity. Molecular analysis at multiple loci revealed insufficient genetic diversity to distinguish terrestrial isolates from strains infecting marine mammals. Furthermore, SnSAG specific primers applied against DNA from the closely related species, Sarcocystis falcatula, lead to the discovery of highly similar orthologs to SnSAG2, 3, and 4, calling into question the specificity of diagnostic tests based on these antigens. The results of this study suggest a population genetic structure for S. neurona similar to that reported for the related parasite, Toxoplasma gondii, dominated by a limited number of successful genotypes. Published by Elsevier B.V.

  15. Selective inhibition of Sarcocystis neurona calcium-dependent protein kinase 1 for equine protozoal myeloencephalitis therapy.

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    Ojo, Kayode K; Dangoudoubiyam, Sriveny; Verma, Shiv K; Scheele, Suzanne; DeRocher, Amy E; Yeargan, Michelle; Choi, Ryan; Smith, Tess R; Rivas, Kasey L; Hulverson, Matthew A; Barrett, Lynn K; Fan, Erkang; Maly, Dustin J; Parsons, Marilyn; Dubey, Jitender P; Howe, Daniel K; Van Voorhis, Wesley C

    2016-12-01

    Sarcocystis neurona is the most frequent cause of equine protozoal myeloencephalitis, a debilitating neurological disease of horses that can be difficult to treat. We identified SnCDPK1, the S. neurona homologue of calcium-dependent protein kinase 1 (CDPK1), a validated drug target in Toxoplasma gondii. SnCDPK1 shares the glycine "gatekeeper" residue of the well-characterized T. gondii enzyme, which allows the latter to be targeted by bumped kinase inhibitors. This study presents detailed molecular and phenotypic evidence that SnCDPK1 can be targeted for rational drug development. Recombinant SnCDPK1 was tested against four bumped kinase inhibitors shown to potently inhibit both T. gondii (Tg) CDPK1 and T. gondii tachyzoite growth. SnCDPK1 was inhibited by low nanomolar concentrations of these BKIs and S. neurona growth was inhibited at 40-120nM concentrations. Thermal shift assays confirmed these bumped kinase inhibitors bind CDPK1 in S. neurona cell lysates. Treatment with bumped kinase inhibitors before or after invasion suggests that bumped kinase inhibitors interfere with S. neurona mammalian host cell invasion in the 0.5-2.5μM range but interfere with intracellular division at 2.5μM. In vivo proof-of-concept experiments were performed in a murine model of S. neurona infection. The experimental infected groups treated for 30days with compound BKI-1553 (n=10 mice) had no signs of disease, while the infected control group had severe signs and symptoms of infection. Elevated antibody responses were found in 100% of control infected animals, but only 20% of BKI-1553 treated infected animals. Parasites were found in brain tissues of 100% of the control infected animals, but only in 10% of the BKI-1553 treated animals. The bumped kinase inhibitors used in these assays have been chemically optimized for potency, selectivity and pharmacokinetic properties, and hence are good candidates for treatment of equine protozoal myeloencephalitis. Copyright © 2016

  16. Exposure of free-living jaguars to Toxoplasma gondii, Neospora caninum and Sarcocystis neurona in the Brazilian Pantanal

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    Selma Samiko Miyazaki Onuma

    2014-12-01

    Full Text Available Toxoplasma gondii, Neospora caninum and Sarcocystis neurona are related apicomplexan parasites that cause reproductive and neurological disorders in a wide range of domestic and wild animals. In the present study, the immunofluorescence antibody test (IFAT was used to investigate the presence of antibodies against T. gondii, N. caninum and S. neurona in the sera of 11 free-living jaguars (Panthera onca in two protected areas in the Pantanal region of Mato Grosso state, Brazil. Ten jaguars (90.9% showed seropositivity for T. gondii, eight (72.7% for S. neurona, and seven (63.6% for N. caninum antigens. Our findings reveal exposure of jaguars to these related coccidian parasites and circulation of these pathogens in this wild ecosystem. To the best of our knowledge, this is the first serological detection of N. caninum and S. neurona in free-living jaguars.

  17. Modest genetic differentiation among North American populations of Sarcocystis neurona may reflect expansion in its geographic range.

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    Sundar, N; Asmundsson, I M; Thomas, N J; Samuel, M D; Dubey, J P; Rosenthal, B M

    2008-03-25

    Sarcocystis neurona is an important cause of neurological disease in horses (equine protozoal myeloencephalitis, EPM) and sea otters in the United States. In addition, EPM-like disease has been diagnosed in several other land and marine mammals. Opossums are its only definitive hosts. Little genetic diversity among isolates of S. neurona from different hosts has been reported. Here, we used 11 microsatellites to characterize S. neurona DNA isolated from natural infections in 22 sea otters (Enhydra lutris) from California and Washington and in 11 raccoons (Procyon lotor) and 1 striped skunk (Mephitis mephitis) from Wisconsin. By jointly analyzing these 34 isolates with 26 isolates previously reported, we determined that geographic barriers may limit S. neurona dispersal and that only a limited subset of possible parasite genotypes may have been introduced to recently established opossum populations. Moreover, our study confirms that diverse intermediate hosts share a common infection source, the opossum (Didelphis virginiana).

  18. Exposure of free-living jaguars to Toxoplasma gondii, Neospora caninum and Sarcocystis neurona in the Brazilian Pantanal.

    Science.gov (United States)

    Onuma, Selma Samiko Miyazaki; Melo, Andréia Lima Tomé; Kantek, Daniel Luis Zanella; Crawshaw-Junior, Peter Gransden; Morato, Ronaldo Gonçalves; May-Júnior, Joares Adenílson; Pacheco, Thábata dos Anjos; Aguiar, Daniel Moura de

    2014-01-01

    Toxoplasma gondii, Neospora caninum and Sarcocystis neurona are related apicomplexan parasites that cause reproductive and neurological disorders in a wide range of domestic and wild animals. In the present study, the immunofluorescence antibody test (IFAT) was used to investigate the presence of antibodies against T. gondii, N. caninum and S. neurona in the sera of 11 free-living jaguars (Panthera onca) in two protected areas in the Pantanal region of Mato Grosso state, Brazil. Ten jaguars (90.9%) showed seropositivity for T. gondii, eight (72.7%) for S. neurona, and seven (63.6%) for N. caninum antigens. Our findings reveal exposure of jaguars to these related coccidian parasites and circulation of these pathogens in this wild ecosystem. To the best of our knowledge, this is the first serological detection of N. caninum and S. neurona in free-living jaguars.

  19. Comparison of prevalence factors in horses with and without seropositivity to Neospora hughesi and/or Sarcocystis neurona.

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    Pusterla, Nicola; Tamez-Trevino, Eva; White, Alexandria; Vangeem, Joshua; Packham, Andrea; Conrad, Patricia A; Kass, Philip

    2014-05-01

    Equine protozoal myeloencephalitis is a commonly diagnosed neurological disease of horses in North America and is caused by infection with Sarcocystis neurona or Neospora hughesi. The aim of this study was to compare prevalence factors among horses seropositive or seronegative to N. hughesi and/or S. neurona. A total of 3123 submissions were included in the study, with horses originating from 49 States. Thirty-eight animals from 21 States tested seropositive for N. hughesi only, 840 horses from 40 States were seropositive for S. neurona only, 25 horses from 14 States were seropositive for both protozoa, and 2220 horses from 49 States tested seronegative for both parasites. Significant associations were found between geographical location (State), month of submission, breed and serological status. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Genetic variation among isolates of Sarcocystis neurona, the agent of protozoal myeloencephalitis, as revealed by amplified fragment length polymorphism markers.

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    Elsheikha, H M; Schott, H C; Mansfield, L S

    2006-06-01

    Sarcocystis neurona causes serious neurological disease in horses and other vertebrates in the Americas. Based on epidemiological data, this parasite has recently emerged. Here, the genetic diversity of Sarcocystis neurona was evaluated using the amplified fragment length polymorphism (AFLP) method. Fifteen S. neurona taxa from different regions collected over the last 10 years were used; six isolates were from clinically diseased horses, eight isolates were from wild-caught opossums (Didelphis virginiana), and one isolate was from a cowbird (Molothrus ater). Additionally, four outgroup taxa were also fingerprinted. Nine primer pairs were used to generate AFLP patterns, with a total number of amplified fragments ranging from 30 to 60, depending on the isolate and primers tested. Based on the presence/absence of amplified AFLP fragments and pairwise similarity values, all the S. neurona isolates tested were clustered in one monophyletic group. No significant correlation could be found between genomic similarity and host origin of the S. neurona isolates. AFLP revealed significant intraspecific genetic variations, and S. neurona appeared as a highly variable species. Furthermore, linkage disequilibrium analysis suggested that S. neurona populations within Michigan have an intermediate type of population structure that includes characteristics of both clonal and panamictic population structures. AFLP is a reliable molecular technique that has provided one of the most informative approaches to ascertain phylogenetic relationships in S. neurona and its closest relatives, allowing them to be clustered by relative similarity using band matching and unweighted pair group method with arithmetic mean analysis, which may be applicable to other related protozoal species.

  1. Phylogenetic congruence of Sarcocystis neurona Dubey et al., 1991 (Apicomplexa: Sarcocystidae) in the United States based on sequence analysis and restriction fragment length polymorphism (RFLP).

    Science.gov (United States)

    Elsheikha, Hany M; Murphy, Alice J; Mansfield, Linda S

    2005-07-01

    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 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.

  2. Frequency of antibodies against Sarcocystis neurona and Neospora caninum in domestic cats in the state of Bahia, Brazil.

    Science.gov (United States)

    Meneses, Iris Daniela Santos de; Andrade, Müller Ribeiro; Uzêda, Rosângela Soares; Bittencourt, Marta Vasconcelos; Lindsay, David Scott; Gondim, Luís Fernando Pita

    2014-01-01

    Sarcocystis neurona is the major agent of equine protozoal myeloencephalitis. It infects several mammalian species in the Americas, where the definitive hosts, marsupials of the genus Didelphis (D. virginiana and D. albiventris) are found. Domestic cats are one of the confirmed intermediate hosts of the parasite; however, antibodies against S. neurona had never before been demonstrated in Brazilian cats. The aim of this study was to determine whether cats in Bahia, Brazil, are exposed to the parasite. A total of 272 feline serum samples (134 from feral and 138 from house cats) were subjected to an indirect fluorescent antibody test using cultured merozoites of S. neurona as antigen. Positivity was detected in 4.0% (11/272) of the tested samples, with titers ranging from 25 to 800. The feline sera were also tested for antibodies against the protozoan Neospora caninum, with an observed antibody frequency of 2.9%. To the author's knowledge, this is the first study to report antibodies against S. neurona in Brazilian cats. We conclude that cats are exposed to the parasite in the region of this study. Further investigations are needed to confirm the role of cats in the transmission cycle of S. neurona in Brazil.

  3. Frequency of antibodies against Sarcocystis neurona and Neospora caninum in domestic cats in the state of Bahia, Brazil

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    Iris Daniela Santos de Meneses

    2014-12-01

    Full Text Available Sarcocystis neurona is the major agent of equine protozoal myeloencephalitis. It infects several mammalian species in the Americas, where the definitive hosts, marsupials of the genus Didelphis (D. virginiana and D. albiventris are found. Domestic cats are one of the confirmed intermediate hosts of the parasite; however, antibodies against S. neurona had never before been demonstrated in Brazilian cats. The aim of this study was to determine whether cats in Bahia, Brazil, are exposed to the parasite. A total of 272 feline serum samples (134 from feral and 138 from house cats were subjected to an indirect fluorescent antibody test using cultured merozoites of S. neurona as antigen. Positivity was detected in 4.0% (11/272 of the tested samples, with titers ranging from 25 to 800. The feline sera were also tested for antibodies against the protozoan Neospora caninum, with an observed antibody frequency of 2.9%. To the author's knowledge, this is the first study to report antibodies against S. neurona in Brazilian cats. We conclude that cats are exposed to the parasite in the region of this study. Further investigations are needed to confirm the role of cats in the transmission cycle of S. neurona in Brazil.

  4. Evidence that Surface Proteins Sn14 and Sn16 of Sarcocystis neurona Merozoites Are Involved in Infection and Immunity†

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    Liang, Fang Ting; Granstrom, David E.; Zhao, Xiao Min; Timoney, John F.

    1998-01-01

    Sarcocystis neurona is the etiologic agent of equine protozoal myeloencephalitis (EPM). Based on an analysis of 25,000 equine serum and cerebrospinal fluid (CSF) samples, including samples from horses with neurologic signs typical of EPM or with histologically or parasitologically confirmed EPM, four major immunoblot band patterns have been identified. Twenty-three serum and CSF samples representing each of the four immunoblot patterns were selected from 220 samples from horses with neurologic signs resembling EPM and examined for inhibitory effects on the infectivity of S. neurona by an in vitro neutralization assay. A high correlation between immunoblot band pattern and neutralizing activity was detected. Two proteins, Sn14 and Sn16 (14 and 16 kDa, respectively), appeared to be important for in vitro infection. A combination of the results of surface protein labeling, immunoprecipitation, Western blotting, and trypsin digestion suggests that these molecules are surface proteins and may be useful components of a vaccine against S. neurona infection. Although S. neurona is an obligate intracellular parasite, it is potentially a target for specific antibodies which may lyse merozoites via complement or inhibit their attachment and penetration to host cells. PMID:9573058

  5. Development and evaluation of a Sarcocystis neurona-specific IgM capture enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Murphy, J E; Marsh, A E; Reed, S M; Meadows, C; Bolten, K; Saville, W J A

    2006-01-01

    Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses caused primarily by the protozoal parasite Sarcocystis neurona. Currently available antemortem diagnostic testing has low specificity. The hypothesis of this study was that serum and cerebrospinal fluid (CSF) of horses experimentally challenged with S neurona would have an increased S neurona-specific IgM (Sn-IgM) concentration after infection, as determined by an IgM capture enzyme linked immunoassay (ELISA). The ELISA was based on the S neurona low molecular weight protein SNUCD-1 antigen and the monoclonal antibody 2G5 labeled with horseradish peroxidase. The test was evaluated using serum and CSF from 12 horses experimentally infected with 1.5 million S neurona sporocysts and 16 horses experimentally infected with varying doses (100 to 100,000) of S neurona sporocysts, for which results of histopathologic examination of the central nervous system were available. For horses challenged with 1.5 million sporocysts, there was a significant increase in serum Sn-IgM concentrations compared with values before infection at weeks 2-6 after inoculation (P neurona, there were significant increases in serum Sn-IgM concentration at various points in time after inoculation, depending on the challenge dose (P < .01). In addition, there was a significant increase between the CSF Sn-IgM concentrations before and after inoculation (P < .0001). These results support further evaluation of the assay as a diagnostic test during the acute phase of EPM.

  6. Seroprevalence of Toxoplasma gondii, Sarcocystis neurona, and Encephalitozoon cuniculi in three species of lemurs from St. Catherines Island, GA, USA.

    Science.gov (United States)

    Yabsley, Michael J; Jordan, Carly N; Mitchell, Sheila M; Norton, Terry M; Lindsay, David S

    2007-03-15

    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.

  7. Prevalence of agglutinating antibodies to Sarcocystis neurona in skunks (Mephitis Mephitis), raccoons (Procyon lotor), and opossums (Didelphis Virginiana) from Connecticut.

    Science.gov (United States)

    Mitchell, Sheila M; Richardson, Dennis J; Cheadle, M Andy; Zajac, Anne M; Lindsay, David S

    2002-10-01

    Equine protozoal myeloencephalitis is the most important protozoan disease of horses in North America and is usually caused by Sarcocystis neurona. Natural cases of encephalitis caused by S. neurona have been reported in skunks (Mephitis mephitis) and raccoons (Procyon lotor). Opossums (Didelphis spp.) are the only known definitive host. Sera from 24 striped skunks, 12 raccoons, and 7 opossums (D. virginiana) from Connecticut were examined for agglutinating antibodies to S. neurona using the S. neurona agglutination test (SAT) employing formalin-fixed merozoites as antigen. The SAT was validated for skunk sera using pre- and postinfection serum samples from 2 experimentally infected skunks. Of the 24 (46%) skunks 11 were positive, and all 12 raccoons were positive for S. neurona antibodies. None of the 7 opossums was positive for antibodies to S. neurona. These results suggest that exposure to sporocysts of S. neurona by intermediate hosts is high in Connecticut. The absence of antibodies in opossums collected from the same areas is most likely because of the absence of systemic infection in the definitive host.

  8. Cross-sectional study of serum antibodies against Sarcocystis neurona in cats tested for antibodies against Toxoplasma gondii.

    Science.gov (United States)

    Rossano, Mary G; Murphy, Alice J; Vrable, Ruth A; Vanzo, Nicole E; Lewis, Stacy K; Sheline, Katherine D; Kaneene, John B; Mansfield, Linda S

    2002-08-15

    To determine apparent seroprevalence of antibodies against Sarcocystis neurona in a population of domestic cats previously tested for antibodies against Toxoplasma gondii. Cross-sectional study. Serum from 196 domestic cats. Banked serum samples submitted to the Michigan State University Animal Health Diagnostic Laboratory for T. gondii diagnostic testing were tested for antibodies against S. neurona by use of an indirect fluorescent antibody (IFA) test and a western blot test. Submission records were analyzed to determine descriptive statistics and test for associations between positive results of a test for S. neurona and other variables in the data set. 10 of 196 (5%) samples yielded positive results for antibodies against S. neurona by use of western blot analysis, whereas 27 samples yielded positive results by use of the IFA. No association was found between S. neurona western blot test results and T. gondii test results, age, sex, or the reason for T. gondii testing. The S. neurona IFA titer was positively and significantly associated with positive results of western blot analysis. Domestic cats are not likely to play a substantial role as intermediate hosts in the natural life cycle of S. neurona. Results indicate that natural infection of domestic cats may occur, and small animal practitioners should be aware of this fact when evaluating cats with neurologic disease. The S. neurona IFA test had lower specificity than western blot analysis.

  9. Phylogenetic relationships of Sarcocystis neurona of horses and opossums to other cyst-forming coccidia deduced from SSU rRNA gene sequences.

    Science.gov (United States)

    Elsheikha, Hany M; Lacher, David W; Mansfield, Linda S

    2005-11-01

    Phylogenetic analyses based on sequences of the nuclear-encoded small subunit rRNA (ssurRNA) gene were performed to examine the origin, phylogeny, and biogeographic relationships of Sarcocystis neurona isolates from opossums and horses from the State of Michigan, USA, in relation to other cyst-forming coccidia. A total of 31 taxa representing all recognized subfamilies and genera of Sarcocystidae were included in the analyses with clonal isolates of two opossum and two horse S. neurona. Phylogenies obtained by the four tree-building methods were consistent with the classical taxonomy based on morphological criteria. The "isosporid" coccidia Neospora, Toxoplasma, Besnoitia, Isospora lacking stieda bodies, and Hyaloklossia formed a sister group to the Sarcocystis spp. Sarcocystis species were divided into three main lineages; S. neurona isolates were located in the second lineage and clustered with S. mucosa, S. dispersa, S. lacertae, S. rodentifelis, S. muris, and Frenkelia spp. Alignment of S. neurona SSU rRNA gene sequences of Michigan opossum isolates (MIOP5, MIOP20) and a S. neurona Michigan horse isolate (MIH8) showed 100% identity. These Michigan isolates differed in 2/1085 bp (0.2%) from a Kentucky S. neurona horse isolate (SN5). Additionally, S. neurona isolates from horses and opossums were identical based on the ultrastructural features and PCR-RFLP analyses thus forming a phylogenetically indistinct group in these regions. These findings revealed the concordance between the morphological and molecular data and confirmed that S. neurona from opossums and horses originated from the same phylogenetic origin.

  10. Genotyping of Toxoplasma gondii and Sarcocystis spp. in road-killed wild mammals from the Central Western Region of the State of São Paulo, Brazil

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    Virgínia Bodelão Richini-Pereira

    Full Text Available Abstract INTRODUCTION: Road-killed wild animals host zoonotic pathogens such as Toxoplasma gondii, offering a new opportunity for the epidemiological study of these infectious organisms. METHODS This investigation aimed to determine the presence of T. gondii and other apicomplexan parasites in tissue samples of 64 road-killed wild animals, using polymerase chain reaction (PCR. Positive samples were then typed by PCR-restriction fragment length polymorphism (RFLP using 7 markers: SAG1, 5′-3′SAG2, SAG3, BTUB, c29-6, PK1, and Apico. PCR-RFLP targeting 18S ribosomal RNA (rRNA genes was also performed on all samples to detect other apicomplexan parasites. RESULTS T. gondii DNA was detected in 16 tissue samples from 8 individual animals, as follows: 1 Cerdocyon thous (crab-eating fox, 1 Didelphis albiventris (white-eared opossum, 1 Lutreolina crassicaudata (lutrine opossum, 2 Myrmecophaga tridactyla (giant anteater, 1 Procyon cancrivorus (crab-eating raccoon, and 2 Sphiggurus spinosus (Paraguay hairy dwarf porcupine. Seven different T. gondii genotypes were identified, 6 of which were novel. Typing by 18S rRNA verified these 16 T. gondii-infected samples, and identified 1 Sarcocystis spp.-infected animal [Dasypus novemcinctus (nine-banded armadillo]. The amplified T. gondii (GenBank accession No. L37415.1 and Sarcocystis spp. 18S rRNA products were confirmed by sequencing. CONCLUSIONS Our results indicate that T. gondii is commonly present in wild mammals, which act as sources of infection for humans and animals, including other wild species. The approach employed herein proved useful for detecting T. gondii and Sarcocystis spp. in the environment and identifying their natural reservoirs, contributing to our understanding of host-parasite interactions.

  11. MANAGEMENT OF ACUTE RENAL FAILURE WITH DELAYED HYPERCALCEMIA SECONDARY TO SARCOCYSTIS NEURONA-INDUCED MYOSITIS AND RHABDOMYOLYSIS IN A CALIFORNIA SEA LION (ZALOPHUS CALIFORNIANUS).

    Science.gov (United States)

    Alexander, Amy B; Hanley, Christopher S; Duncan, Mary C; Ulmer, Kyle; Padilla, Luis R

    2015-09-01

    A 3-yr-old captive-born California sea lion (Zalophus californianus) developed Sarcocystis neurona-induced myositis and rhabdomyolysis that led to acute renal failure. The sea lion was successfully managed with fluid therapy, antiprotozoals, antibiotics, anti-inflammatories, antiemetics, gastroprotectants, and diuretics, but developed severe delayed hypercalcemia, a syndrome identified in humans after traumatic or exertion-induced rhabdomyolysis. Treatment with calcitonin was added to the management, and the individual recovered fully. The case emphasizes that animals with rhabdomyolysis-induced renal failure risk developing delayed hypercalcemia, which may be life threatening, and calcium levels should be closely monitored past the resolution of renal failure.

  12. Anti-Sarcocystis neurona immunostaining associated with equine protozoal myeloencephalitis in Brazil Imunomarcação de Sarcocystis neurona associada com um caso de mieloencefalite protozoária eqüina no Brasil

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    Tatiane Alves da Paixão

    2007-12-01

    Full Text Available A retrospective study (1942 to 2005 of histopathological lesions included samples of central nervous system (SNC from 203 animals in the Equidae family. A total of 42.4% of these samples had significant pathological changes, which were classified as inflammatory (62.8%, degenerative (25.6%, circulatory (10.5%, and neoplasic (1.1% lesions. Immunohistochemistry anti-Sarcocystis neurona antigens was performed in all the cases with inflammatory changes (54, of which one of the case of encephalitis resulted positive to immunostaining. Although evidence of EPM (Equine Protozoal Myeloencephalitis has been previously reported in Brazil, to the best of our knowledge, this is the first report in which characteristic EPM lesion was associated with anti-S. neurona immunostaining in Brazil.Em um estudo retrospectivo (de 1942 a 2005, amostras do sistema nervoso central de 203 eqüídeos foram avaliadas para a presença de alterações histológicas. Dessas amostras, 42,4% apresentaram alguma lesão histopatológica significativa, das quais foram classificadas como alterações inflamatórias (62,8%, degenerativas (25,6%, circulatórias (10,5% e neoplásicas (1,1%. Fragmentos de SNC dos 54 animais com alterações inflamatórias foram avaliados para detecção de antígenos de Sarocystis neurona pela técnica de imunoistoquímica, que foi positiva em um caso de encefalite em eqüino. Embora haja registros de MPE no Brasil, este é o primeiro caso confirmado imunoistoquimicamente.

  13. Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii.

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    Jered M Wendte

    Full Text Available Tissue-encysting coccidia, including Toxoplasma gondii and Sarcocystis neurona, are heterogamous parasites with sexual and asexual life stages in definitive and intermediate hosts, respectively. During its sexual life stage, T. gondii reproduces either by genetic out-crossing or via clonal amplification of a single strain through self-mating. Out-crossing has been experimentally verified as a potent mechanism capable of producing offspring possessing a range of adaptive and virulence potentials. In contrast, selfing and other life history traits, such as asexual expansion of tissue-cysts by oral transmission among intermediate hosts, have been proposed to explain the genetic basis for the clonal population structure of T. gondii. In this study, we investigated the contributing roles self-mating and sexual recombination play in nature to maintain clonal population structures and produce or expand parasite clones capable of causing disease epidemics for two tissue encysting parasites. We applied high-resolution genotyping against strains isolated from a T. gondii waterborne outbreak that caused symptomatic disease in 155 immune-competent people in Brazil and a S. neurona outbreak that resulted in a mass mortality event in Southern sea otters. In both cases, a single, genetically distinct clone was found infecting outbreak-exposed individuals. Furthermore, the T. gondii outbreak clone was one of several apparently recombinant progeny recovered from the local environment. Since oocysts or sporocysts were the infectious form implicated in each outbreak, the expansion of the epidemic clone can be explained by self-mating. The results also show that out-crossing preceded selfing to produce the virulent T. gondii clone. For the tissue encysting coccidia, self-mating exists as a key adaptation potentiating the epidemic expansion and transmission of newly emerged parasite clones that can profoundly shape parasite population genetic structures or cause

  14. Seroprevalences of anti-Sarcocystis neurona and anti-Neospora hughesi antibodies among healthy equids in the United States.

    Science.gov (United States)

    James, Kaitlyn E; Smith, Woutrina A; Conrad, Patricia A; Packham, Andrea E; Guerrero, Leopoldo; Ng, Mitchell; Pusterla, Nicola

    2017-06-01

    OBJECTIVE To describe the general seroprevalence of anti-Sarcocystis neurona and anti-Neospora hughesi antibodies among healthy equids by use of indirect fluorescent antibody tests and determine potential risk factors for seropositivity. DESIGN Cross-sectional study. SAMPLE Whole blood samples collected from 5,250 equids (1 sample/animal) across 18 states in the United States during October 2013. PROCEDURES Information regarding potential risk factors (geographic region, breed, primary use, sex, and age) was collected along with the blood samples. For each equid, an indirect fluorescent antibody test was used to determine serum titers of antibody against each of the 2 protozoal parasites. Mixed-effects logistic regression models were created to determine ORs for seropositivity. RESULTS The overall seroprevalence of anti-S neurona and anti-N hughesi antibodies in the tested equids was 78% and 34%, respectively. Of the equids, 31% were seropositive and 18% were seronegative for antibodies against both parasites. Factors associated with equids being seropositive for anti-S neurona antibodies were residence in the South, warmblood breed, and age > 5 years. Seroprevalence of anti-N hughesi antibodies did not differ among equids in different states across the country, but warmblood breed and age > 5 years were associated with seropositivity. CONCLUSIONS AND CLINICAL RELEVANCE With regard to risk factors for S neurona and N hughesi exposure and antibody response among tested equids, older age was not unexpected; however, the influences of warmblood breed and geographic location on seropositivity for anti-S neurona antibody but not for anti-N hughesi antibody deserve further investigation.

  15. Sarcocystis neurona-specific immunoglobulin G in the serum and cerebrospinal fluid of horses administered S neurona vaccine.

    Science.gov (United States)

    Witonsky, Sharon; Morrow, Jennifer K; Leger, Clare; Dascanio, John; Buechner-Maxwell, Virginia; Palmer, Wally; Kline, Kristen; Cook, Anne

    2004-01-01

    A vaccine against Sarcocystis neurona, which induces equine protozoal myeloencephalitis (EPM), has received conditional licensure in the United States. A major concern is whether the immunoglobulin G (IgG) response elicited by the vaccine will compromise the use of Western blotting (WB) as a diagnostic tool in vaccinated horses with neurologic disease. Our goals were to determine if vaccination (1) causes seroconversion: (2) causes at least a transient increase in S neurona-specific IgG in the cerebrospinal fluid (CSF); and (3) induces an IgG response that can be differentiated from that induced by natural exposure. Horses included in the study (n = 29) were older than 6 months with no evidence of neurologic disease. The presence or absence of anti-S neurona antibodies in the serum of each horse was determined by WB analysis. Seropositive horses had CSF collected and submitted for cytology, CSF index, and WB analysis. The vaccine was administered to all the horses and boostered 3-4 weeks later. On day 14 after the 2nd administration, serum and CSF were collected and analyzed. Eighty-nine percent (8 of 9) of the initial seronegative horses seroconverted after vaccination, of which 57% (4 of 7) had anti-S neurona IgG in their CSE Eighty percent (16 of 20) of the seropositive horses had an increase in serum S neurona IgG after vaccination. Of the 6 of 20 horses that were initially seropositive/CSF negative, 2 were borderline positive for anti-S neurona IgG in the CSF, 2 tested positive, and 2 were excluded because the CSF sample had been contaminated by blood. There were no WB banding patterns that distinguished samples from horses that seroconverted due to vaccination versus natural exposure. Caution must be used in interpreting WB analysis from neurologic horses that have been recently vaccinated for EPM.

  16. Diagnosis and treatment of Sarcocystis neurona-induced myositis in a free-ranging California sea lion.

    Science.gov (United States)

    Carlson-Bremer, Daphne P; Gulland, Frances M D; Johnson, Christine K; Colegrove, Kathleen M; Van Bonn, William G

    2012-02-01

    An underweight, lethargic adult female California sea lion (Zalophus californianus) became stranded along the California shore and was captured and transported to a rehabilitation hospital for assessment and care. Initial physical assessment revealed the sea lion was lethargic and in poor body condition. Active myositis was diagnosed on the basis of concurrent elevations in activities of alanine aminotransferase and creatine kinase detected during serum biochemical analysis. Infection with Sarcocystis neurona was diagnosed after serologic titers increased 4-fold over a 3-week period. Diagnosis was confirmed on the basis of histopathologic findings, positive results on immunohistochemical staining, and results of quantitative PCR assay on biopsy specimens obtained from the diaphragm and muscles of the dorsal cervical region. Anticoccidial treatment was instituted with ponazuril (10 mg/kg [4.5 mg/lb], PO, q 24 h) and continued for 28 days. Prednisone (0.2 mg/kg [0.09 mg/lb], PO, q 12 h) was administered for 2 days and then every 24 hours for 5 days to treat associated inflammation. At the end of treatment, the sea lion was clinically normal, alanine aminotransferase and creatine kinase values were within reference limits, and antibody titers against S neurona had decreased 6-fold. The sea lion was released approximately 3 months after becoming stranded. S neurona-induced myositis was diagnosed in a free-ranging California sea lion. On the basis of the successful treatment and release of this sea lion, anticoccidial treatment should be considered for marine mammals in which protozoal disease is diagnosed.

  17. Prevalence of antibodies to Neospora caninum, Sarcocystis neurona, and Toxoplasma gondii in wild horses from central Wyoming.

    Science.gov (United States)

    Dubey, J P; Mitchell, S M; Morrow, J K; Rhyan, J C; Stewart, L M; Granstrom, D E; Romand, S; Thulliez, P; Saville, W J; Lindsay, D S

    2003-08-01

    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.

  18. Self-mating in the definitive host potentiates clonal outbreaks of the apicomplexan parasites Sarcocystis neurona and Toxoplasma gondii.

    Science.gov (United States)

    Wendte, Jered M; Miller, Melissa A; Lambourn, Dyanna M; Magargal, Spencer L; Jessup, David A; Grigg, Michael E

    2010-12-23

    Tissue-encysting coccidia, including Toxoplasma gondii and Sarcocystis neurona, are heterogamous parasites with sexual and asexual life stages in definitive and intermediate hosts, respectively. During its sexual life stage, T. gondii reproduces either by genetic out-crossing or via clonal amplification of a single strain through self-mating. Out-crossing has been experimentally verified as a potent mechanism capable of producing offspring possessing a range of adaptive and virulence potentials. In contrast, selfing and other life history traits, such as asexual expansion of tissue-cysts by oral transmission among intermediate hosts, have been proposed to explain the genetic basis for the clonal population structure of T. gondii. In this study, we investigated the contributing roles self-mating and sexual recombination play in nature to maintain clonal population structures and produce or expand parasite clones capable of causing disease epidemics for two tissue encysting parasites. We applied high-resolution genotyping against strains isolated from a T. gondii waterborne outbreak that caused symptomatic disease in 155 immune-competent people in Brazil and a S. neurona outbreak that resulted in a mass mortality event in Southern sea otters. In both cases, a single, genetically distinct clone was found infecting outbreak-exposed individuals. Furthermore, the T. gondii outbreak clone was one of several apparently recombinant progeny recovered from the local environment. Since oocysts or sporocysts were the infectious form implicated in each outbreak, the expansion of the epidemic clone can be explained by self-mating. The results also show that out-crossing preceded selfing to produce the virulent T. gondii clone. For the tissue encysting coccidia, self-mating exists as a key adaptation potentiating the epidemic expansion and transmission of newly emerged parasite clones that can profoundly shape parasite population genetic structures or cause devastating disease

  19. Analysis of the Sarcocystis neurona microneme protein SnMIC10: protein characteristics and expression during intracellular development.

    Science.gov (United States)

    Hoane, Jessica S; Carruthers, Vernon B; Striepen, Boris; Morrison, David P; Entzeroth, Rolf; Howe, Daniel K

    2003-07-01

    Sarcocystis neurona, an apicomplexan parasite, is the primary causative agent of equine protozoal myeloencephalitis. Like other members of the Apicomplexa, S. neurona zoites possess secretory organelles that contain proteins necessary for host cell invasion and intracellular survival. From a collection of S. neurona expressed sequence tags, we identified a sequence encoding a putative microneme protein based on similarity to Toxoplasma gondii MIC10 (TgMIC10). Pairwise sequence alignments of SnMIC10 to TgMIC10 and NcMIC10 from Neospora caninum revealed approximately 33% identity to both orthologues. The open reading frame of the S. neurona gene encodes a 255 amino acid protein with a predicted 39-residue signal peptide. Like TgMIC10 and NcMIC10, SnMIC10 is predicted to be hydrophilic, highly alpha-helical in structure, and devoid of identifiable adhesive domains. Antibodies raised against recombinant SnMIC10 recognised a protein band with an apparent molecular weight of 24 kDa in Western blots of S. neurona merozoites, consistent with the size predicted for SnMIC10. In vitro secretion assays demonstrated that this protein is secreted by extracellular merozoites in a temperature-dependent manner. Indirect immunofluorescence analysis of SnMIC10 showed a polar labelling pattern, which is consistent with the apical position of the micronemes, and immunoelectron microscopy provided definitive localisation of the protein to these secretory organelles. Further analysis of SnMIC10 in intracellular parasites revealed that expression of this protein is temporally regulated during endopolygeny, supporting the view that micronemes are only needed during host cell invasion. Collectively, the data indicate that SnMIC10 is a microneme protein that is part of the excreted/secreted antigen fraction of S. neurona. Identification and characterisation of additional S. neurona microneme antigens and comparisons to orthologues in other Apicomplexa could provide further insight into the

  20. 化隆县家畜膈肌住肉孢子虫感染情况研究%Study on the Infection of Sarcocystis in Diaphragmatic Muscle of Yak and Sheep in Hualong County

    Institute of Scientific and Technical Information of China (English)

    康明; 李英; 石文辉; 席进桂花

    2013-01-01

    [目的]摸清化隆县牦牛和绵羊住肉孢子虫的感染情况.[方法]采用膈肌压片镜检法,对青海省化隆县6个乡(镇)的牦牛和绵羊膈肌中住肉孢子虫的感染情况进行研究.[结果]化隆县牦牛和绵羊膈肌中住肉孢子虫的感染率分别为40%和93.3%,感染寄生强度分别为11和41 个/g肉样.化隆县牦牛和绵羊中住肉孢子虫的感染情况较为严重.[结论]该研究可为今后住肉孢子虫的防治提供科学依据.%[Objective] The research aimed to investigate the infection situation of Sarcocystis in yak and sheep in Hualong County. [Method] Using microscope examination method of pressed diaphragmatic muscle, the infection situations of Sarcocystis in diaphragmatic muscle of yak and sheep in six towns (countrysides) of Hualong County were studied. [Result] The infection rate of Sarcocystis in diaphragm muscle of yak and sheep in Hualong County were 40% and 93.3% respectively. And the intensity of infection were 11 and 41 worms in each gram of meat sample. The infection of Sarcocystis in yak and sheep was serious in Hualong County. [Conclusion] The research could provide scientific basis for the control of Sarcocystis in future.

  1. Improved detection of equine antibodies against Sarcocystis neurona using polyvalent ELISAs based on the parasite SnSAG surface antigens.

    Science.gov (United States)

    Yeargan, Michelle R; Howe, Daniel K

    2011-02-28

    Equine protozoal myeloencephalitis (EPM) is a common neurologic disease of horses that is caused by the apicomplexan pathogen Sarcocystis neurona. To help improve serologic diagnosis of S. neurona infection, we have modified existing enzyme-linked immunosorbent assays (ELISAs) based on the immunogenic parasite surface antigens SnSAG2, SnSAG3, and SnSAG4 to make the assays polyvalent, thereby circumventing difficulties associated with parasite antigenic variants and diversity in equine immune responses. Two approaches were utilized to achieve polyvalence: (1) mixtures of the individual recombinant SnSAGs (rSnSAGs) were included in single ELISAs; (2) a collection of unique SnSAG chimeras that fused protein domains from different SnSAG surface antigens into a single recombinant protein were generated for use in the ELISAs. These new assays were assessed using a defined sample set of equine sera and cerebrospinal fluids (CSFs) that had been characterized by Western blot and/or were from confirmed EPM horses. While all of the polyvalent ELISAs performed relatively well, the highest sensitivity and specificity (100%/100%) were achieved with assays containing the rSnSAG4/2 chimera (Domain 1 of SnSAG4 fused to SnSAG2) or using a mixture of rSnSAG3 and rSnSAG4. The rSnSAG4 antigen alone and the rSnSAG4/3 chimera (Domain 1 of SnSAG4 fused to Domain 2 of SnSAG3) exhibited the next best accuracy at 95.2% sensitivity and 100% specificity. Binding ratios and percent positivity (PP) ratios, determined by comparing the mean values for positive versus negative samples, showed that the most advantageous signal to noise ratios were provided by rSnSAG4 and the rSnSAG4/3 chimera. Collectively, our results imply that a polyvalent ELISA based on SnSAG4 and SnSAG3, whether as a cocktail of two proteins or as a single chimeric protein, can give optimal results in serologic testing of serum or CSF for the presence of antibodies against S. neurona. The use of polyvalent SnSAG ELISAs will

  2. Prevalence of antibodies to Trypanosoma cruzi, Leishmania infantum, Encephalitozoon cuniculi, Sarcocystis neurona, and Neospora caninum in Capybara, Hydrochoerus hydrochaeris, from São Paulo State, Brazil.

    Science.gov (United States)

    Valadas, Samantha; Gennari, Solange Maria; Yai, Lucia Eiko Oishi; Rosypal, Alexa C; Lindsay, David S

    2010-06-01

    Little is known about the importance of capybara, Hydrochoerus hydrochaeris, as reservoirs for parasites of zoonotic or veterinary importance. Sera from 63 capybaras, from 6 counties in the state of São Paulo, Brazil, were examined for antibodies to Trypanosoma cruzi, Leishmania infantum, Encephalitozoon cuniculi, Sarcocystis neurona, and Neospora caninum using an indirect immunofluorescent antibody test. Five (8%) of the 63 capybaras had antibodies to T. cruzi epimastigotes. None of the samples from capybara reacted positively with L. infantum promastigotes or with spores of E. cuniculi . Two (3%) of the serum samples were positive for antibodies to S. neurona merozoites, and 2 (3%) of the serum samples were positive for antibodies to N. caninum tachyzoites. A serum sample from 1 capybara was positive for antibodies to both T. cruzi and N. caninum. None of the remaining 62 samples reacted with more than 1 parasite.

  3. Ultrastructural and molecular confirmation of the development of Sarcocystis neurona tissue cysts in the central nervous system of southern sea otters (Enhydra lutris nereis).

    Science.gov (United States)

    Miller, M A; Barr, B C; Nordhausen, R; James, E R; Magargal, S L; Murray, M; Conrad, P A; Toy-Choutka, S; Jessup, D A; Grigg, M E

    2009-10-01

    In 2004, three wild sea otters were diagnosed with putative Sarcocystis neurona-associated meningoencephalitis by histopathology and immunohistochemistry. Schizonts, free merozoites and tissue cysts were observed in the brains of all three infected animals. Tissue cysts walls from sea otter 1 (SO1) stained positively using anti-S. neurona polyclonal antiserum. However, positive staining does not preclude infection by closely related or cross-reactive tissue cyst-forming coccidian parasites. Two immature tissue cysts in the brain of SO1 were examined using transmission electron microscopy. Ultrastructural features included cyst walls with thin villous projections up to 1 microm long with tapered ends and a distinctive, electron-dense outer lining layer composed of linearly-arranged, semi-circular structures with a "hobnailed" surface contour. Small numbers of microtubules extended down through the villi into the underlying granular layer. Metrocytes were short and plump with an anterior apical complex, 22 sub-pellicular microtubules, numerous free ribosomes and no rhoptries. Some metrocytes appeared to be dividing, with two adjacent nuclear profiles. Collectively these ultrastructural features were compatible with developing protozoal cysts and were similar to prior descriptions of S. neurona tissue cysts. Panspecific 18S rDNA primers were utilized to identify protozoa infecting the brains of these otters and DNA amplification and additional sequencing at the ITS1 locus confirmed that all three otters were infected with S. neurona. No other Sarcocystis spp. were detected in the brains or skeletal muscles of these animals by immunohistochemistry or PCR. We believe this is the first ultrastructural and molecular confirmation of the development of S. neurona tissue cysts in the CNS of any animal.

  4. SnSAG5 is an alternative surface antigen of Sarcocystis neurona strains that is mutually exclusive to SnSAG1.

    Science.gov (United States)

    Crowdus, Carolyn A; Marsh, Antoinette E; Saville, Willliam J; Lindsay, David S; Dubey, J P; Granstrom, David E; Howe, Daniel K

    2008-11-25

    Sarcocystis neurona is an obligate intracellular parasite that causes equine protozoal myeloencephalitis (EPM). Previous work has identified a gene family of paralogous surface antigens in S. neurona called SnSAGs. These surface proteins are immunogenic in their host animals, and are therefore candidate molecules for development of diagnostics and vaccines. However, SnSAG diversity exists in strains of S. neurona, including the absence of the major surface antigen gene SnSAG1. Instead, sequence for an alternative SnSAG has been revealed in two of the SnSAG1-deficient strains. Herein, we present data characterizing this new surface protein, which we have designated SnSAG5. The results indicated that the protein encoded by the SnSAG5 sequence is indeed a surface-associated molecule that has characteristics consistent with the other SAGs identified in S. neurona and related parasites. Importantly, Western blot analyses of a collection of S. neurona strains demonstrated that 6 of 13 parasite isolates express SnSAG5 as a dominant surface protein instead of SnSAG1. Conversely, SnSAG5 was not detected in SnSAG1-positive strains. One strain, which was isolated from the brain of a sea otter, did not express either SnSAG1 or SnSAG5. Genetic analysis with SnSAG5-specific primers confirmed the presence of the SnSAG5 gene in Western blot-positive strains, while also suggesting the presence of a novel SnSAG sequence in the SnSAG1-deficient, SnSAG5-deficient otter isolate. The findings provide further indication of S. neurona strain diversity, which has implications for diagnostic testing and development of vaccines against EPM as well as the population biology of Sarcocystis cycling in the opossum definitive host.

  5. Strains of Sarcocystis neurona exhibit differences in their surface antigens, including the absence of the major surface antigen SnSAG1.

    Science.gov (United States)

    Howe, Daniel K; Gaji, Rajshekhar Y; Marsh, Antoinette E; Patil, Bhagyashree A; Saville, William J; Lindsay, David S; Dubey, J P; Granstrom, David E

    2008-05-01

    A gene family of surface antigens is expressed by merozoites of Sarcocystis neurona, the primary cause of equine protozoal myeloencephalitis (EPM). These surface proteins, designated SnSAGs, are immunodominant and therefore excellent candidates for development of EPM diagnostics or vaccines. Prior work had identified an EPM isolate lacking the major surface antigen SnSAG1, thus suggesting there may be some diversity in the SnSAGs expressed by different S. neurona isolates. Therefore, a bioinformatic, molecular and immunological study was conducted to assess conservation of the SnSAGs. Examination of an expressed sequence tag (EST) database revealed several notable SnSAG polymorphisms. In particular, the EST information implied that the EPM strain SN4 lacked the major surface antigen SnSAG1. The absence of this surface antigen from the SN4 strain was confirmed by both Western blot and Southern blot. To evaluate SnSAG polymorphisms in the S. neurona population, 14 strains were examined by Western blots using monospecific polyclonal antibodies against the four described SnSAGs. The results of these analyses demonstrated that SnSAG2, SnSAG3, and SnSAG4 are present in all 14 S. neurona strains tested, although some variance in SnSAG4 was observed. Importantly, SnSAG1 was not detected in seven of the strains, which included isolates from four cases of EPM and a case of fatal meningoencephalitis in a sea otter. Genetic analyses by PCR using gene-specific primers confirmed the absence of the SnSAG1 locus in six of these seven strains. Collectively, the data indicated that there is heterogeneity in the surface antigen composition of different S. neurona isolates, which is an important consideration for development of serological tests and prospective vaccines for EPM. Furthermore, the diversity reported herein likely extends to other phenotypes, such as strain virulence, and may have implications for the phylogeny of the various Sarcocystis spp. that undergo sexual stages

  6. Seroepidemiology of Sarcocystis neurona, Toxoplasma gondii and Neospora spp. among horses in the south of the state of Minas Gerais, Brazil

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    Manoel Junqueira Maciel Ribeiro

    2016-01-01

    Full Text Available Abstract The present study used the indirect fluorescent antibody test (IFAT to determine the seroprevalence of Sarcocystis neurona, Toxoplasma gondii and Neospora spp., and evaluated the variables associated with these infections among 506 apparently healthy horses, reared in the south of the state of Minas Gerais, Brazil. This study was conducted between April 2012 and October 2013. Among the horses, the true prevalence of S. neurona was 26% (95% CI: 22.0-30.4%, T. gondii 19.9% (95% CI: 15.5-24.8% and Neospora spp. 23.9% (95% CI: 19.9-28.1%; and among the farms, 88.3% (95% CI: 74.4-91.6%, 71.6% (95% CI: 41-92.8% and 85% (95% CI: 70.7-96.1%, respectively. Regarding mixed infection, 17 horses (3.4% were seropositive for both S. neurona and T. gondii, 16 (3.2% for T. gondii and Neospora spp. and 14 (2.8% for S. neurona and Neospora spp. The associations between seropositivity and variables relating to the structure of the farm, management and health were analyzed using the logistic regression analysis, through the generalized estimating equations (GEE. The results suggest that the south of Minas Gerais is an enzootic area for S. neurona, T. gondii and Neospora spp. among horses, with prevalence of asymptomatic subclinical or chronic infections.

  7. Dual congenital transmission of Toxoplasma gondii and Sarcocystis neurona in a late-term aborted pup from a chronically infected southern sea otter (Enhydra lutris nereis).

    Science.gov (United States)

    Shapiro, Karen; Miller, Melissa A; Packham, Andrea E; Aguilar, Beatriz; Conrad, Patricia A; Vanwormer, Elizabeth; Murray, Michael J

    2016-03-01

    Toxoplasma gondii and Sarcocystis neurona are protozoan parasites with terrestrial definitive hosts, and both pathogens can cause fatal disease in a wide range of marine animals. Close monitoring of threatened southern sea otters (Enhydra lutris nereis) in California allowed for the diagnosis of dual transplacental transmission of T. gondii and S. neurona in a wild female otter that was chronically infected with both parasites. Congenital infection resulted in late-term abortion due to disseminated toxoplasmosis. Toxoplasma gondii and S. neurona DNA was amplified from placental tissue culture, as well as from fetal lung tissue. Molecular characterization of T. gondii revealed a Type X genotype in isolates derived from placenta and fetal brain, as well as in all tested fetal organs (brain, lung, spleen, liver and thymus). This report provides the first evidence for transplacental transmission of T. gondii in a chronically infected wild sea otter, and the first molecular and immunohistochemical confirmation of concurrent transplacental transmission of T. gondii and S. neurona in any species. Repeated fetal and/or neonatal losses in the sea otter dam also suggested that T. gondii has the potential to reduce fecundity in chronically infected marine mammals through parasite recrudescence and repeated fetal infection.

  8. Effects of blood contamination of cerebrospinal fluid on results of indirect fluorescent antibody tests for detection of antibodies against Sarcocystis neurona and Neospora hughesi.

    Science.gov (United States)

    Finno, Carrie J; Packham, Andrea E; David Wilson, W; Gardner, Ian A; Conrad, Patricia A; Pusterla, Nicola

    2007-05-01

    The purpose of this study was to determine the effect of blood contamination of cerebrospinal fluid (CSF) on the results of indirect fluorescent antibody tests (IFATs) for Sarcocystis neurona and Neospora hughesi. The in vitro study used antibody-negative CSF collected from non-neurologic horses immediately after euthanasia and blood samples from 40 healthy horses that had a range of IFAT antibody titers against S. neurona and N. hughesi. Serial dilutions of whole blood were made in seronegative CSF to generate blood-contaminated CSF with red blood cell (RBC) concentrations ranging from 10 to 100,000 RBCs/microl. The blood-contaminated CSF samples were then tested for antibodies against both pathogens using IFAT. Blood contamination of CSF had no detectable effect on IFAT results for S. neurona or N. hughesi at any serologic titer when the RBC concentration in CSF was or=5) for S. neurona and N. hughesi were detected only when the corresponding serum titers were >or=160 and >or=80, respectively. The IFAT performed on CSF is reliable for testing horses for equine protozoal myeloencephalitis caused by S. neurona or N. hughesi, even when blood contamination causes the RBC concentration in CSF to be up to 10,000 RBCs/microl.

  9. Purine salvage in the apicomplexan Sarcocystis neurona, and generation of hypoxanthine-xanthine-guanine phosphoribosyltransferase-deficient clones for positive-negative selection of transgenic parasites.

    Science.gov (United States)

    Dangoudoubiyam, Sriveny; Zhang, Zijing; Howe, Daniel K

    2014-09-01

    Sarcocystis neurona is an apicomplexan parasite that causes severe neurological disease in horses and marine mammals. The Apicomplexa are all obligate intracellular parasites that lack purine biosynthesis pathways and rely on the host cell for their purine requirements. Hypoxanthine-xanthine-guanine phosphoribosyltransferase (HXGPRT) and adenosine kinase (AK) are key enzymes that function in two complementary purine salvage pathways in apicomplexans. Bioinformatic searches of the S. neurona genome revealed genes encoding HXGPRT, AK and all of the major purine salvage enzymes except purine nucleoside phosphorylase. Wild-type S. neurona were able to grow in the presence of mycophenolic acid (MPA) but were inhibited by 6-thioxanthine (6-TX), suggesting that the pathways involving either HXGPRT or AK are functional in this parasite. Prior work with Toxoplasma gondii demonstrated the utility of HXGPRT as a positive-negative selection marker. To enable the use of HXGPRT in S. neurona, the SnHXGPRT gene sequence was determined and a gene-targeting plasmid was transfected into S. neurona. SnHXGPRT-deficient mutants were selected with 6-TX, and single-cell clones were obtained. These Sn∆HXG parasites were susceptible to MPA and could be complemented using the heterologous T. gondii HXGPRT gene. In summary, S. neurona possesses both purine salvage pathways described in apicomplexans, thus allowing the use of HXGPRT as a positive-negative drug selection marker in this parasite.

  10. Seroepidemiology of Sarcocystis neurona, Toxoplasma gondii and Neospora spp. among horses in the south of the state of Minas Gerais, Brazil.

    Science.gov (United States)

    Ribeiro, Manoel Junqueira Maciel; Rosa, Marina Helena Figueredo; Bruhn, Fábio Raphael Pascoti; Garcia, Adriana de Mello; Rocha, Christiane Maria Barcellos Magalhães da; Guimarães, Antônio Marcos

    2016-06-07

    The present study used the indirect fluorescent antibody test (IFAT) to determine the seroprevalence of Sarcocystis neurona, Toxoplasma gondii and Neospora spp., and evaluated the variables associated with these infections among 506 apparently healthy horses, reared in the south of the state of Minas Gerais, Brazil. This study was conducted between April 2012 and October 2013. Among the horses, the true prevalence of S. neurona was 26% (95% CI: 22.0-30.4%), T. gondii 19.9% (95% CI: 15.5-24.8%) and Neospora spp. 23.9% (95% CI: 19.9-28.1%); and among the farms, 88.3% (95% CI: 74.4-91.6%), 71.6% (95% CI: 41-92.8%) and 85% (95% CI: 70.7-96.1%), respectively. Regarding mixed infection, 17 horses (3.4%) were seropositive for both S. neurona and T. gondii, 16 (3.2%) for T. gondii and Neospora spp. and 14 (2.8%) for S. neurona and Neospora spp. The associations between seropositivity and variables relating to the structure of the farm, management and health were analyzed using the logistic regression analysis, through the generalized estimating equations (GEE). The results suggest that the south of Minas Gerais is an enzootic area for S. neurona, T. gondii and Neospora spp. among horses, with prevalence of asymptomatic subclinical or chronic infections.

  11. Prevalence of antibodies to Trypanosoma cruzi, Toxoplasma gondii, Encephalitozoon cuniculi, Sarcocystis neurona, Besnoitia darlingi, and Neospora caninum in North American opossums, Didelphis virginiana, from southern Louisiana.

    Science.gov (United States)

    Houk, Alice E; Goodwin, David G; Zajac, Anne M; Barr, Stephen C; Dubey, J P; Lindsay, David S

    2010-12-01

    We examined the prevalence of antibodies to zoonotic protozoan parasites ( Trypanosoma cruzi, Toxoplasma gondii, and Encephalitozoon cuniculi) and protozoans of veterinary importance ( Neospora caninum, Sarcocystis neurona, and Besnoitia darlingi) in a population of North American opossums ( Didelphis virginiana) from Louisiana. Samples from 30 opossums were collected as part of a survey for T. cruzi in Louisiana. Frozen sera from these 30 opossums were examined using an indirect immunofluorescent antibody test (IFAT) against in vitro-produced antigenic stages of these protozoans. Additionally, 24 of the 30 samples were examined using hemoculture, and all 30 were examined in the modified direct agglutination test (MAT) for antibodies to To. gondii. The prevalences of reactive IFAT samples were as follows: 60% for T. cruzi, 27% for To. gondii, 23% for E. cuniculi, 17% for S. neurona, 47% for B. darlingi, and 0% for N. caninum. Hemoculture revealed that 16 (67%) of 24 samples were positive for T. cruzi, compared to 18 of 30 (60%) by IFAT. The sensitivity and specificity for the IFAT compared to hemoculture was 100% for each. The modified direct agglutination test revealed that 9 (30%) of the 30 samples from opossums had antibodies to To. gondii , compared to 8 (27%) using the IFAT. The sensitivity and specificity of the IFAT compared to the MAT was 100% and 72%, respectively.

  12. The heptanucleotide motif GAGACGC is a key component of a cis-acting promoter element that is critical for SnSAG1 expression in Sarcocystis neurona.

    Science.gov (United States)

    Gaji, Rajshekhar Y; Howe, Daniel K

    2009-07-01

    The apicomplexan parasite Sarcocystis neurona undergoes a complex process of intracellular development, during which many genes are temporally regulated. The described study was undertaken to begin identifying the basic promoter elements that control gene expression in S. neurona. Sequence analysis of the 5'-flanking region of five S. neurona genes revealed a conserved heptanucleotide motif GAGACGC that is similar to the WGAGACG motif described upstream of multiple genes in Toxoplasma gondii. The promoter region for the major surface antigen gene SnSAG1, which contains three heptanucleotide motifs within 135 bases of the transcription start site, was dissected by functional analysis using a dual luciferase reporter assay. These analyses revealed that a minimal promoter fragment containing all three motifs was sufficient to drive reporter molecule expression, with the presence and orientation of the 5'-most heptanucleotide motif being absolutely critical for promoter function. Further studies should help to identify additional sequence elements important for promoter function and for controlling gene expression during intracellular development by this apicomplexan pathogen.

  13. A novel Sarcocystis neurona genotype XIII is associated with severe encephalitis in an unexpectedly broad range of marine mammals from the northeastern Pacific Ocean.

    Science.gov (United States)

    Barbosa, Lorraine; Johnson, Christine K; Lambourn, Dyanna M; Gibson, Amanda K; Haman, Katherine H; Huggins, Jessica L; Sweeny, Amy R; Sundar, Natarajan; Raverty, Stephen A; Grigg, Michael E

    2015-08-01

    Sarcocystis neurona is an important cause of protozoal encephalitis among marine mammals in the northeastern Pacific Ocean. To characterise the genetic type of S. neurona in this region, samples from 227 stranded marine mammals, most with clinical or pathological evidence of protozoal disease, were tested for the presence of coccidian parasites using a nested PCR assay. The frequency of S. neurona infection was 60% (136/227) among pinnipeds and cetaceans, including seven marine mammal species not previously known to be susceptible to infection by this parasite. Eight S. neurona fetal infections identified this coccidian parasite as capable of being transmitted transplacentally. Thirty-seven S. neurona-positive samples were multilocus sequence genotyped using three genetic markers: SnSAG1-5-6, SnSAG3 and SnSAG4. A novel genotype, referred to as Type XIII within the S. neurona population genetic structure, has emerged recently in the northeastern Pacific Ocean and is significantly associated with an increased severity of protozoal encephalitis and mortality among multiple stranded marine mammal species. Published by Elsevier Ltd.

  14. [Control of toxicity of Sarcocystis fayeri in horsemeat by freezing treatment and prevention of food poisoning caused by raw consumption of horsemeat].

    Science.gov (United States)

    Harada, Seiya; Furukawa, Masato; Tokuoka, Eisuke; Matsumoto, Kazutoshi; Yahiro, Shunsuke; Miyasaka, Jiro; Saito, Morihiro; Kamata, Yoichi; Watanabe, Maiko; Irikura, Daisuke; Matsumoto, Hiroshi; Sugita-Konishi, Yoshiko

    2013-01-01

    More than 27 outbreaks per year of food poisoning caused by consuming horse meat were reported in Kumamoto Prefecture (including Kumamoto City) from January 2009 to September 2011. It was found that the causative agent of the outbreaks was a protein with a molecular weight of 15 kDa that had originated from bradyzoites of Sarcocystis fayeri parasitizing the horse meat. Rabit ileal loop tests showed that pepsin treatment of homogenates of frozen horse meat containing the cysts of S. fayeri induced loss of toxicity, presumably by digestion of the proteinous causative agent(s). Slices of horse meat containing the cysts were frozen at below -20°C for various periods. The cysts were collected after thawing the slices, then treated in an artificial stomach juice containing pepsin. The bradyzoites of the cysts kept at -20°C for 48 hr or more completely disappeared. Simultaneously, the 15 kDa protein also disappeared in the frozen cysts. After notifying the public and recommending freezing treatment of horse meat, no subsequent cases of food poisoning were reported. This indicates that freezing of horse meat is effective to prevent the occurrence of food poisoning caused by consuming raw horse meat containing S. fayeri.

  15. Sarcocystis neurona merozoites express a family of immunogenic surface antigens that are orthologues of the Toxoplasma gondii surface antigens (SAGs) and SAG-related sequences.

    Science.gov (United States)

    Howe, Daniel K; Gaji, Rajshekhar Y; Mroz-Barrett, Meaghan; Gubbels, Marc-Jan; Striepen, Boris; Stamper, Shelby

    2005-02-01

    Sarcocystis neurona is a member of the Apicomplexa that causes myelitis and encephalitis in horses but normally cycles between the opossum and small mammals. Analysis of an S. neurona expressed sequence tag (EST) database revealed four paralogous proteins that exhibit clear homology to the family of surface antigens (SAGs) and SAG-related sequences of Toxoplasma gondii. The primary peptide sequences of the S. neurona proteins are consistent with the two-domain structure that has been described for the T. gondii SAGs, and each was predicted to have an amino-terminal signal peptide and a carboxyl-terminal glycolipid anchor addition site, suggesting surface localization. All four proteins were confirmed to be membrane associated and displayed on the surface of S. neurona merozoites. Due to their surface localization and homology to T. gondii surface antigens, these S. neurona proteins were designated SnSAG1, SnSAG2, SnSAG3, and SnSAG4. Consistent with their homology, the SnSAGs elicited a robust immune response in infected and immunized animals, and their conserved structure further suggests that the SnSAGs similarly serve as adhesins for attachment to host cells. Whether the S. neurona SAG family is as extensive as the T. gondii SAG family remains unresolved, but it is probable that additional SnSAGs will be revealed as more S. neurona ESTs are generated. The existence of an SnSAG family in S. neurona indicates that expression of multiple related surface antigens is not unique to the ubiquitous organism T. gondii. Instead, the SAG gene family is a common trait that presumably has an essential, conserved function(s).

  16. The SnSAG merozoite surface antigens of Sarcocystis neurona are expressed differentially during the bradyzoite and sporozoite life cycle stages.

    Science.gov (United States)

    Gautam, A; Dubey, J P; Saville, W J; Howe, D K

    2011-12-29

    Sarcocystis neurona is a two-host coccidian parasite whose complex life cycle progresses through multiple developmental stages differing at morphological and molecular levels. The S. neurona merozoite surface is covered by multiple, related glycosylphosphatidylinositol-linked proteins, which are orthologous to the surface antigen (SAG)/SAG1-related sequence (SRS) gene family of Toxoplasma gondii. Expression of the SAG/SRS proteins in T. gondii and another related parasite Neospora caninum is life-cycle stage specific and seems necessary for parasite transmission and persistence of infection. In the present study, the expression of S. neurona merozoite surface antigens (SnSAGs) was evaluated in the sporozoite and bradyzoite stages. Western blot analysis was used to compare SnSAG expression in merozoites versus sporozoites, while immunocytochemistry was performed to examine expression of the SnSAGs in merozoites versus bradyzoites. These analyses revealed that SnSAG2, SnSAG3 and SnSAG4 are expressed in sporozoites, while SnSAG5 was appeared to be downregulated in this life cycle stage. In S. neurona bradyzoites, it was found that SnSAG2, SnSAG3, SnSAG4 and SnSAG5 were either absent or expression was greatly reduced. As shown for T. gondii, stage-specific expression of the SnSAGs may be important for the parasite to progress through its developmental stages and complete its life cycle successfully. Thus, it is possible that the SAG switching mechanism by these parasites could be exploited as a point of intervention. As well, the alterations in surface antigen expression during different life cycle stages may need to be considered when designing prospective approaches for protective vaccination. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. A protozoal-associated epizootic impacting marine wildlife: mass-mortality of southern sea otters (Enhydra lutris nereis) due to Sarcocystis neurona infection.

    Science.gov (United States)

    Miller, Melissa A; Conrad, Patricia A; Harris, Michael; Hatfield, Brian; Langlois, Gregg; Jessup, David A; Magargal, Spencer L; Packham, Andrea E; Toy-Choutka, Sharon; Melli, Ann C; Murray, Michael A; Gulland, Frances M; Grigg, Michael E

    2010-09-20

    During April 2004, 40 sick and dead southern sea otters (Enhydra lutris nereis) were recovered over 18km of coastline near Morro Bay, California. This event represented the single largest monthly spike in mortality ever recorded during 30 years of southern sea otter stranding data collection. Because of the point-source nature of the event and clinical signs consistent with severe, acute neurological disease, exposure to a chemical or marine toxin was initially considered. However, detailed postmortem examinations revealed lesions consistent with an infectious etiology, and further investigation confirmed the protozoan parasite Sarcocystis neurona as the underlying cause. Tissues from 94% of examined otters were PCR-positive for S. neurona, based on DNA amplification and sequencing at the ITS-1 locus, and 100% of tested animals (n=14) had elevated IgM and IgG titers to S. neurona. Evidence to support the point-source character of this event include the striking spatial and temporal clustering of cases and detection of high concentrations of anti-S. neurona IgM in serum of stranded animals. Concurrent exposure to the marine biotoxin domoic acid may have enhanced susceptibility of affected otters to S. neurona and exacerbated the neurological signs exhibited by stranded animals. Other factors that may have contributed to the severity of this epizootic include a large rainstorm that preceded the event and an abundance of razor clams near local beaches, attracting numerous otters close to shore within the affected area. This is the first report of a localized epizootic in marine wildlife caused by apicomplexan protozoa.

  18. Prevalence of and risk factors associated with the presence of Sarcocystis neurona sporocysts in opossum (Didelphis virginiana) from Michigan: a retrospective study.

    Science.gov (United States)

    Elsheikha, Hany M; Murphy, Alice J; Mansfield, Linda S

    2004-11-10

    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.

  19. Sarcocystis neurona Merozoites Express a Family of Immunogenic Surface Antigens That Are Orthologues of the Toxoplasma gondii Surface Antigens (SAGs) and SAG-Related Sequences†

    Science.gov (United States)

    Howe, Daniel K.; Gaji, Rajshekhar Y.; Mroz-Barrett, Meaghan; Gubbels, Marc-Jan; Striepen, Boris; Stamper, Shelby

    2005-01-01

    Sarcocystis neurona is a member of the Apicomplexa that causes myelitis and encephalitis in horses but normally cycles between the opossum and small mammals. Analysis of an S. neurona expressed sequence tag (EST) database revealed four paralogous proteins that exhibit clear homology to the family of surface antigens (SAGs) and SAG-related sequences of Toxoplasma gondii. The primary peptide sequences of the S. neurona proteins are consistent with the two-domain structure that has been described for the T. gondii SAGs, and each was predicted to have an amino-terminal signal peptide and a carboxyl-terminal glycolipid anchor addition site, suggesting surface localization. All four proteins were confirmed to be membrane associated and displayed on the surface of S. neurona merozoites. Due to their surface localization and homology to T. gondii surface antigens, these S. neurona proteins were designated SnSAG1, SnSAG2, SnSAG3, and SnSAG4. Consistent with their homology, the SnSAGs elicited a robust immune response in infected and immunized animals, and their conserved structure further suggests that the SnSAGs similarly serve as adhesins for attachment to host cells. Whether the S. neurona SAG family is as extensive as the T. gondii SAG family remains unresolved, but it is probable that additional SnSAGs will be revealed as more S. neurona ESTs are generated. The existence of an SnSAG family in S. neurona indicates that expression of multiple related surface antigens is not unique to the ubiquitous organism T. gondii. Instead, the SAG gene family is a common trait that presumably has an essential, conserved function(s). PMID:15664946

  20. Evidence that antibodies against recombinant SnSAG1 of Sarcocystis neurona merozoites are involved in infection and immunity in equine protozoal myeloencephalitis.

    Science.gov (United States)

    Ellison, Siobhan; Witonsky, Sharon

    2009-07-01

    Sarcocystis neurona is the principal etiologic agent of equine protozoal myeloencephalitis (EPM). An immunodominant protein of S. neurona, SnSAG-1, is expressed by the majority of S. neurona merozoites isolated from spinal tissues of horses diagnosed with EPM and may be a candidate for diagnostic tests and prophylaxis for EPM. Five horses were vaccinated with adjuvanted recombinant SnSAG1 (rSnSAG1) and 5 control (sham vaccinated) horses were vaccinated with adjuvant only. Serum was evaluated pre- and post-vaccination, prior to challenge, for antibodies against rSnSAG1 and inhibitory effects on the infectivity of S. neurona by an in vitro serum neutralization assay. The effect of vaccination with rSnSAG1 on in vivo infection by S. neurona was evaluated by challenging all the horses with S. neurona merozoites. Blinded daily examinations and 4 blinded neurological examinations were used to evaluate the presence of clinical signs of EPM. The 5 vaccinated horses developed serum and cerebrospinal fluid (CSF) titers of SnSAG1, detected by enzyme-linked immunosorbent assay (ELISA), post-vaccination. Post-vaccination serum from vaccinated horses was found to have an inhibitory effect on merozoites, demonstrated by in vitro bioassay. Following the challenge, the 5 control horses displayed clinical signs of EPM, including ataxia. While 4 of the 5 vaccinated horses did not become ataxic. One rSnSAG-1 vaccinated horse showed paresis in 1 limb with muscle atrophy. All horses showed mild, transient, cranial nerve deficits; however, disease did not progress to ataxia in rSnSAG-1 vaccinated horses. The study showed that vaccination with rSnSAG-1 produced antibodies in horses that neutralized merozoites when tested by in vitro culture and significantly reduced clinical signs demonstrated by in vivo challenge.

  1. A protozoal-associated epizootic impacting marine wildlife: Mass-mortality of southern sea otters (Enhydra lutris nereis) due to Sarcocystis neurona infection

    Science.gov (United States)

    Miller, M.A.; Conrad, P.A.; Harris, M.; Hatfield, B.; Langlois, G.; Jessup, David A.; Magargal, S.L.; Packham, A.E.; Toy-Choutka, S.; Melli, A.C.; Murray, M.A.; Gulland, F.M.; Grigg, M.E.

    2010-01-01

    During April 2004, 40 sick and dead southern sea otters (Enhydra lutris nereis) were recovered over 18 km of coastline near Morro Bay, California. This event represented the single largest monthly spike in mortality ever recorded during 30 years of southern sea otter stranding data collection. Because of the point-source nature of the event and clinical signs consistent with severe, acute neurological disease, exposure to a chemical or marine toxin was initially considered. However, detailed postmortem examinations revealed lesions consistent with an infectious etiology, and further investigation confirmed the protozoan parasite Sarcocystis neurona as the underlying cause. Tissues from 94% of examined otters were PCR-positive for S. neurona, based on DNA amplification and sequencing at the ITS-1 locus, and 100% of tested animals (n= 14) had elevated IgM and IgG titers to S. neurona. Evidence to support the point-source character of this event include the striking spatial and temporal clustering of cases and detection of high concentrations of anti- S. neurona IgM in serum of stranded animals. Concurrent exposure to the marine biotoxin domoic acid may have enhanced susceptibility of affected otters to S. neurona and exacerbated the neurological signs exhibited by stranded animals. Other factors that may have contributed to the severity of this epizootic include a large rainstorm that preceded the event and an abundance of razor clams near local beaches, attracting numerous otters close to shore within the affected area. This is the first report of a localized epizootic in marine wildlife caused by apicomplexan protozoa. ?? 2010 Elsevier B.V.

  2. First identification of Sarcocystis tenella (Railliet, 1886) Moulé, 1886 (Protozoa: Apicomplexa) by PCR in naturally infected sheep from Brazil.

    Science.gov (United States)

    da Silva, Rodrigo Costa; Su, Chunlei; Langoni, Helio

    2009-11-12

    Sarcocystis tenella is a dog-sheep protozoan parasite, causing a widespread enzootic muscle parasitosis and neurological disease mainly in lambs. This parasite is pathogenic to sheep and important to the economical production of sheep. The present study was initially aimed to determine Toxoplasma gondii infection and the occurrence of co-infection with other Apicomplexa parasites in 602 Brazilian sheep. Twenty of these sheep were positive with antibodies to T. gondii by MAT and IFAT-IgG tests, positive with PCR-RFLP genotyping at multiple loci, and parasites were isolated from mice infected with sheep tissue samples. Two additional sheep born in Brazil, a 2-year-old female Polwarth (Ideal) sheep, a breed originated from Australia (#1), and a 1-year-old male Corriedale sheep, a breed originated from New Zealand and Australia (#2) were positive to T. gondii antibodies by serum tests, and PCR, but negative for bioassay in mice. In genotyping at 12 loci, sheep #1 sample and #2 presented positive results only for some markers. PCR-RFLP of 18S ribosomal RNA (18S rRNA) was performed in all 22 animals to identify the possibility of co-infection of T. gondii with other Apicomplexa parasites, such as S. tenella, Neospora caninum and Hammondia hammondi, resulting in a T. gondii profile for the first 20 animals and a unique genotyping profile for sheep #1 and #2, identical to S. tenella. The 18S rRNA PCR products (approximately 310 bp) were sequenced and blasted to GenBank database at NCBI. Both samples were identical to S. tenella 18S rRNA gene (GenBank accession number L24383-1). These results suggest the existence of co-infection of S. tenella with T. gondii in ewes from Brazil.

  3. Inhibitory Activities of Epidermal Growth Factor Receptor Tyrosine Kinase-Targeted Dihydroxyisoflavone and Trihydroxydeoxybenzoin Derivatives on Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum Development

    Science.gov (United States)

    Gargala, G.; Baishanbo, A.; Favennec, L.; François, A.; Ballet, J. J.; Rossignol, J.-F.

    2005-01-01

    Several gene sequences of parasitic protozoa belonging to protein kinase gene families and epidermal growth factor (EGF)-like peptides, which act via binding to receptor tyrosine kinases of the EGF receptor (EGFR) family, appear to mediate host-protozoan interactions. As a clue to EGFR protein tyrosine kinase (PTK) mediation and a novel approach for identifying anticoccidial agents, activities against Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum grown in BM and HCT-8 cell cultures of 52 EGFR PTK inhibitor isoflavone analogs (dihydroxyisoflavone and trihydroxydeoxybenzoine derivatives) were investigated. Their cytotoxicities against host cells were either absent, mild, or moderate by a nitroblue tetrazolium test. At concentrations ranging from 5 to 10 μg/ml, 20 and 5 analogs, including RM-6427 and RM-6428, exhibited an in vitro inhibitory effect of ≥95% against at least one parasite or against all three, respectively. In immunosuppressed Cryptosporidium parvum-infected Mongolian gerbils orally treated with either 200 or 400 mg of agent RM-6427/kg of body weight/day for 8 days, fecal microscopic oocyst shedding was abolished in 6/10 animals (P of 0.05, respectively). After RM-6427 therapy (200 mg/kg/day for 8 days), the reduction in the ratio of animals with intracellular parasites was nearly significant in ileum (P = 0.067) and more marked in the biliary tract (P < 0.0013) than after nitazoxanide or paromomycin treatment (0.05 < P < 0.004). RM-6428 treatment at a regimen of 400 mg/kg/day for 12 days inhibited oocyst shedding, measured using flow cytometry from day 4 (P < 0.05) to day 12 (P < 0.02) of therapy, when 2/15 animals had no shedding (P < 0.0001) and 11/15 were free of gut and/or biliary tract parasites (P < 0.01). No mucosal alteration was microscopically observed for treated or untreated infected gerbils. To our knowledge, this report is the first to suggest that the isoflavone class of agents has the potential for

  4. A new trivalent SnSAG surface antigen chimera for efficient detection of antibodies against Sarcocystis neurona and diagnosis of equine protozoal myeloencephalitis.

    Science.gov (United States)

    Yeargan, Michelle; de Assis Rocha, Izabela; Morrow, Jennifer; Graves, Amy; Reed, Stephen M; Howe, Daniel K

    2015-05-01

    Enzyme-linked immunosorbent assays (ELISAs) based on the SnSAG surface antigens of Sarcocystis neurona provide reliable detection of infection by the parasite. Moreover, accurate serodiagnosis of equine protozoal myeloencephalitis (EPM) is achieved with the SnSAG ELISAs by measuring antibodies in serum and cerebrospinal fluid (CSF) to reveal active infection in the central nervous system. Two independent ELISAs based on recombinant (r)SnSAG2 or a chimeric fusion of SnSAG3 and SnSAG4 (rSnSAG4/3) are currently used together for EPM serodiagnosis to overcome varied antibody responses in different horses. To achieve reliable antibody detection with a single ELISA instead of 2 separate ELISAs, rSnSAG2 was fused with rSnSAG4/3 into a single trivalent protein, designated rSnSAG2/4/3. Paired serum and CSF from 163 horses were tested with all 3 ELISAs. When the consensus antibody titers obtained with the rSnSAG2 and rSnSAG4/3 ELISAs were compared to the single SAG2/4/3 ELISA titers, Spearman rank correlation coefficients of ρ = 0.74 and ρ = 0.90 were obtained for serum and CSF, respectively, indicating strong agreement between the tests. When the rSnSAG2 and rSnSAG4/3 consensus serum-to-CSF titer ratio was compared to the rSnSAG2/4/3 serum-to-CSF titer ratio, the Spearman correlation coefficient was ρ = 0.87, again signifying strong agreement. Importantly, comparing the diagnostic interpretation of the serum-to-CSF titer ratios yielded a Cohen kappa value of 0.77. These findings suggest that the single ELISA based on the trivalent rSnSAG2/4/3 will provide serologic and diagnostic results that are highly comparable to the consensus of the 2 independent ELISAs based on rSnSAG2 and rSnSAG4/3. © 2015 The Author(s).

  5. Epidemiology of Sarcocystis neurona infections in domestic cats (Felis domesticus) and its association with equine protozoal myeloencephalitis (EPM) case farms and feral cats from a mobile spay and neuter clinic.

    Science.gov (United States)

    Stanek, J F; Stich, R W; Dubey, J P; Reed, S M; Njoku, C J; Lindsay, D S; Schmall, L M; Johnson, G K; LaFave, B M; Saville, W J A

    2003-11-28

    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.

  6. Sarcocystis neurona infection in gamma interferon gene knockout (KO) mice: comparative infectivity of sporocysts in two strains of KO mice, effect of trypsin digestion on merozoite viability, and infectivity of bradyzoites to KO mice and cell culture.

    Science.gov (United States)

    Dubey, J P; Sundar, N; Kwok, O C H; Saville, W J A

    2013-09-01

    The protozoan Sarcocystis neurona is the primary cause of Equine Protozoal Myeloencephalitis (EPM). EPM or EPM-like illness has been reported in horses, sea otters, and several other mammals. The gamma interferon gene knockout (KO) mouse is often used as a model to study biology and discovery of new therapies against S. neurona because it is difficult to induce clinical EPM in other hosts, including horses. In the present study, infectivity of three life cycle stages (merozoites, bradyzoites, sporozoites) to KO mice and cell culture was studied. Two strains of KO mice (C57-black, and BALB/c-derived, referred here as black or white) were inoculated orally graded doses of S. neurona sporocysts; 12 sporocysts were infective to both strains of mice and all infected mice died or became ill within 70 days post-inoculation. Although there was no difference in infectivity of sporocysts to the two strains of KO mice, the disease was more severe in black mice. S. neurona bradyzoites were not infectious to KO mice and cell culture. S. neurona merozoites survived 120 min incubation in 0.25% trypsin, indicating that trypsin digestion can be used to recover S. neurona from tissues of acutely infected animals. Published by Elsevier B.V.

  7. Accurate antemortem diagnosis of equine protozoal myeloencephalitis (EPM) based on detecting intrathecal antibodies against Sarcocystis neurona using the SnSAG2 and SnSAG4/3 ELISAs.

    Science.gov (United States)

    Reed, S M; Howe, D K; Morrow, J K; Graves, A; Yeargan, M R; Johnson, A L; MacKay, R J; Furr, M; Saville, W J A; Williams, N M

    2013-01-01

    Recent work demonstrated the value of antigen-specific antibody indices (AI and C-value) to detect intrathecal antibody production against Sarcocystis neurona for antemortem diagnosis of equine protozoal myeloencephalitis (EPM). The study was conducted to assess whether the antigen-specific antibody indices can be reduced to a simple serum : cerebrospinal fluid (CSF) titer ratio to achieve accurate EPM diagnosis. Paired serum and CSF samples from 128 horses diagnosed by postmortem examination. The sample set included 44 EPM cases, 35 cervical-vertebral malformation (CVM) cases, 39 neurologic cases other than EPM or CVM, and 10 non-neurologic cases. Antibodies against S. neurona were measured in serum and CSF pairs using the SnSAG2 and SnSAG4/3 (SnSAG2, 4/3) ELISAs, and the ratio of each respective serum titer to CSF titer was determined. Likelihood ratios and diagnostic sensitivity and specificity were calculated based on serum titers, CSF titers, and serum : CSF titer ratios. Excellent diagnostic sensitivity and specificity was obtained from the SnSAG2, 4/3 serum : CSF titer ratio. Sensitivity and specificity of 93.2 and 81.1%, respectively, were achieved using a ratio cutoff of ≤100, whereas sensitivity and specificity were 86.4 and 95.9%, respectively, if a more rigorous cutoff of ≤50 was used. Antibody titers in CSF also provided good diagnostic accuracy. Serum antibody titers alone yielded much lower sensitivity and specificity. The study confirms the value of detecting intrathecal antibody production for antemortem diagnosis of EPM, and they further show that the antigen-specific antibody indices can be reduced in practice to a simple serum : CSF titer ratio. Copyright © 2013 by the American College of Veterinary Internal Medicine.

  8. First report of human intestinal sarcocystosis in Cambodia.

    Science.gov (United States)

    Khieu, Virak; Marti, Hanspeter; Chhay, Saomony; Char, Meng Chuor; Muth, Sinuon; Odermatt, Peter

    2017-10-01

    Human intestinal sarcocystosis (HIS), caused by Sarcocystis species, is acquired by eating undercooked meat from sarcocyst-containing cattle (S. hominis, S. heydorni) and pigs (S. suihominis). We report on the detection of human intestinal Sarcocystis infections in a cross-sectional survey of Strongyloides stercoralis in early 2014, in Rovieng District, Preah Vihear Province, northern Cambodia. Among 1081 participants, 108 (10.0%) were diagnosed with Sarcocystis spp. oocysts in stool samples. Males had a significantly higher risk of infection than females (OR: 1.9, 95% CI: 1.3-2.9, p=0.001). None of the reported symptoms (abdominal discomfort, diarrhea, muscle pain and itching skin) occurring in the two weeks preceding the examinations were associated with a Sarcocystis infection. Many Sarcocystis cases were found among those who had participated in a wedding celebration and Chinese New Year festivities, where they had consumed raw or insufficiently cooked beef (83.3%) and pork (38.9%) based dishes. This report documents the first HIS cases in Cambodia. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Human and animal sarcocystosis in Malaysia:A review

    Institute of Scientific and Technical Information of China (English)

    Baha Latif; Azdayanti Muslim

    2016-01-01

    Sarcocystosis is a zoonotic disease caused by a coccidian intracellular protozoan parasite of the genus Sarcocystis. More than 200 Sarcocystis species have been recorded and the parasites are found in mammals, birds and reptiles. They require two hosts to complete their life cycle. In Malaysia, sarcocystosis was reported as a potential emerging food and water-borne disease after a series of large outbreak of human infections. There was not enough attention given before even though it was reported in both humans and animals. The first human case of invasive muscular sarcocystosis among local Malaysian was reported in 1975. Besides, a retrospective autopsy examination on 100 tongues revealed 21% positive cases. On top of that, a sero-epidemiological survey conducted in 243 subjects in West Malaysia showed that 19.7% had Sarcocystis antibodies. The clinical symptoms of muscular sarcocystosis were first described comprehensively in 1999. Meanwhile, many types of animals including livestock were found harbor the sarcocysts in their tissue. The first case of human intestinal sarcocystosis was reported in 2014. This review indicates that human sarcocystosis is currently endemic in Malaysia and parallel to that reported in animals. However, more studies and investigations need to be conducted since the source of human infection remains unknown.

  10. Acute muscular sarcocystosis: an international investigation among ill travelers returning from Tioman Island, Malaysia, 2011-2012.

    Science.gov (United States)

    Esposito, Douglas H; Stich, August; Epelboin, Loïc; Malvy, Denis; Han, Pauline V; Bottieau, Emmanuel; da Silva, Alexandre; Zanger, Philipp; Slesak, Günther; van Genderen, Perry J J; Rosenthal, Benjamin M; Cramer, Jakob P; Visser, Leo G; Muñoz, José; Drew, Clifton P; Goldsmith, Cynthia S; Steiner, Florian; Wagner, Noémie; Grobusch, Martin P; Plier, D Adam; Tappe, Dennis; Sotir, Mark J; Brown, Clive; Brunette, Gary W; Fayer, Ronald; von Sonnenburg, Frank; Neumayr, Andreas; Kozarsky, Phyllis E

    2014-11-15

    Through 2 international traveler-focused surveillance networks (GeoSentinel and TropNet), we identified and investigated a large outbreak of acute muscular sarcocystosis (AMS), a rarely reported zoonosis caused by a protozoan parasite of the genus Sarcocystis, associated with travel to Tioman Island, Malaysia, during 2011-2012. Clinicians reporting patients with suspected AMS to GeoSentinel submitted demographic, clinical, itinerary, and exposure data. We defined a probable case as travel to Tioman Island after 1 March 2011, eosinophilia (>5%), clinical or laboratory-supported myositis, and negative trichinellosis serology. Case confirmation required histologic observation of sarcocysts or isolation of Sarcocystis species DNA from muscle biopsy. Sixty-eight patients met the case definition (62 probable and 6 confirmed). All but 2 resided in Europe; all were tourists and traveled mostly during the summer months. The most frequent symptoms reported were myalgia (100%), fatigue (91%), fever (82%), headache (59%), and arthralgia (29%); onset clustered during 2 distinct periods: "early" during the second and "late" during the sixth week after departure from the island. Blood eosinophilia and elevated serum creatinine phosphokinase (CPK) levels were observed beginning during the fifth week after departure. Sarcocystis nesbitti DNA was recovered from 1 muscle biopsy. Clinicians evaluating travelers returning ill from Malaysia with myalgia, with or without fever, should consider AMS, noting the apparent biphasic aspect of the disease, the later onset of elevated CPK and eosinophilia, and the possibility for relapses. The exact source of infection among travelers to Tioman Island remains unclear but needs to be determined to prevent future illnesses. Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  11. Acute Muscular Sarcocystosis: An International Investigation Among Ill Travelers Returning From Tioman Island, Malaysia, 2011–2012

    Science.gov (United States)

    Esposito, Douglas H.; Stich, August; Epelboin, Loïc; Malvy, Denis; Han, Pauline V.; Bottieau, Emmanuel; da Silva, Alexandre; Zanger, Philipp; Slesak, Günther; van Genderen, Perry J. J.; Rosenthal, Benjamin M.; Cramer, Jakob P.; Visser, Leo G.; Muñoz, José; Drew, Clifton P.; Goldsmith, Cynthia S.; Steiner, Florian; Wagner, Noémie; Grobusch, Martin P.; Plier, D. Adam; Tappe, Dennis; Sotir, Mark J.; Brown, Clive; Brunette, Gary W.; Fayer, Ronald; von Sonnenburg, Frank; Neumayr, Andreas; Kozarsky, Phyllis E.

    2015-01-01

    Background Through 2 international traveler-focused surveillance networks (GeoSentinel and TropNet), we identified and investigated a large outbreak of acute muscular sarcocystosis (AMS), a rarely reported zoonosis caused by a protozoan parasite of the genus Sarcocystis, associated with travel to Tioman Island, Malaysia, during 2011–2012. Methods Clinicians reporting patients with suspected AMS to GeoSentinel submitted demographic, clinical, itinerary, and exposure data. We defined a probable case as travel to Tioman Island after 1 March 2011, eosinophilia (>5%), clinical or laboratory-supported myositis, and negative trichinellosis serology. Case confirmation required histologic observation of sarcocysts or isolation of Sarcocystis species DNA from muscle biopsy. Results Sixty-eight patients met the case definition (62 probable and 6 confirmed). All but 2 resided in Europe; all were tourists and traveled mostly during the summer months. The most frequent symptoms reported were myalgia (100%), fatigue (91%), fever (82%), headache (59%), and arthralgia (29%); onset clustered during 2 distinct periods: “early” during the second and “late” during the sixth week after departure from the island. Blood eosinophilia and elevated serum creatinine phosphokinase (CPK) levels were observed beginning during the fifth week after departure. Sarcocystis nesbitti DNA was recovered from 1 muscle biopsy. Conclusions Clinicians evaluating travelers returning ill from Malaysia with myalgia, with or without fever, should consider AMS, noting the apparent biphasic aspect of the disease, the later onset of elevated CPK and eosinophilia, and the possibility for relapses. The exact source of infection among travelers to Tioman Island remains unclear but needs to be determined to prevent future illnesses. PMID:25091309

  12. A presumptive case of Baylisascaris procyonis in a feral green-cheeked Amazon parrot (Amazona viridigenalis).

    Science.gov (United States)

    Done, Lisa B; Tamura, Yoko

    2014-03-01

    A feral green-cheeked Amazon parrot (Amazona viridigenalis), also known as the red-crowned Amazon, with generalized neurologic symptoms was found in Pasadena in Southern California and brought in for treatment. The bird was refractory to a wide variety of medications and supportive treatment. Tests for polyoma virus, psittacine beak and feather disease virus, and West Nile virus as well as Chlamydophila psittaci were negative. Hospitalized and home care continued for a total of 69 days. The bird was rehospitalized on day 66 for increasing severity of clinical signs and found 3 days later hanging with its head down, in respiratory arrest. Resuscitation was unsuccessful. There were no gross pathologic lesions. Histopathology showed a focal subcutaneous fungal caseous granuloma under the skin of the dorsum. Many sarcocysts morphologically consistent with Sarcocystis falcatula were found in the cytoplasm of the skeletal myofibers from skeletal muscles of different locations of this bird, a finding that was considered an incidental, clinically nonsignificant finding in this case. Necrosis with microscopic lesions typical of Baylisascaris spp. neural larva migrans was in the brain. Although multiple histologic serial sections of the brain were examined and a brain squash performed and analyzed, no Baylisascaris larvae were found. This is the first presumptive case of Baylisascaris in a feral psittacine.

  13. Prevalence of agglutinating antibodies to Toxoplasma gondii and Sarcocystis neurona in beavers (Castor canadensis) from Massachusetts

    Science.gov (United States)

    Jordan, C.N.; Kaur, T.; Koenen, K.; DeStefano, S.; Zajac, A.M.; Lindsay, D.S.

    2005-01-01

    The present study examined the seroprevalence of Toxoplasma gondii and Sarcocystls 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. ?? American Society of Pathologists 2005.

  14. Detection of Neospora caninum in tissue sections using a murine monoclonal antibody.

    Science.gov (United States)

    Cole, R A; Lindsay, D S; Dubey, J P; Blagburn, B L

    1993-10-01

    A murine monoclonal antibody (MAb 6G7), isotype IgG2a, produced against tachyzoites of Neospora caninum (isolate NC-1) reacted specifically with tachyzoites of N. caninum in an indirect immunofluorescent antibody test. MAb 6G7 did not react with tachyzoites of Toxoplasma gondii, sporozoites of Isospora suis, Eimeria bovis, or E. tenella, or merozoites of E. bovis in the indirect immunofluorescent antibody test. MAb 6G7 reacted positively with both tachyzoites and bradyzoites of N. caninum in an avidin-biotin peroxidase complex immunohistochemical test on formalin-fixed paraffin-embedded tissues. No reaction was observed with the following: tachyzoites and bradyzoites of T. gondii, T. gondii-like parasites, or Hammondia hammondi; bradyzoites of Frenkelia microti; schizonts and merozoites of Sarcocystis-like organisms; schizonts, sarcocysts, and oocysts/sporocysts of S. cruzi; schizonts and merozoites of S. canis; schizonts of S. hirsuta, S. tenella, and S. capracanis; merozoites of S. neurona and S. neurona-like organisms, E. bovis, or Haemoproteus sp.; bradyzoites and merozoites of S. montanaensis; bradyzoites of S. odocoileocanis, S. cruzi, and S. tenella; meronts, sexual stages, and caryocysts of Caryospora sp. and C. bigenetica; micromerozoites, macromerozoites, and schizonts of Hepatozoon canis; sporozoites, sexual stages, and oocysts of Cryptosporidium parvum and C. baileyi; trophozoites of Monocystis lumbrici, Tritrichomonas foetus, and Balantidium coli; tissue cysts and bradyzoites of Besnoitia sp. and B. jellisoni; amastigotes of Leishmania sp.; and trophic theronts of Ichthyopthirius multifilis. MAb 6G7 reacted with tachyzoites and bradyzoites of N. caninum in natural and experimental infections in dogs, cattle, mice, rats, sheep, and goats, indicating that host origin of the tissues did not affect the performance of the test.

  15. Prevalence of Muscular Sarcosporidiosis in Slaughtered Domestic Pigs in Perak, Peninsular Malaysia

    Science.gov (United States)

    Zainalabidin, Fazly Ann; Noorazmi, Muhamad Syamsul Naim; Bakri, Wan Normaziah Wan Omar; Sathaya, Geethamalar; Ismail, Mohd Iswadi

    2017-01-01

    Sarcosporidiosis is a disease caused by intracellular protozoan parasites, namely, Sarcocystis spp. In pigs, three species of Sarcocystis spp. have been recognised, including Sarcocystis meischeriana, Sarcocystis porcifelis and Sarcocystis suihominis. The aim of this study is to determine the prevalence of muscular sarcosporidiosis in pigs using the pepsin digestion technique. A total of 150 fresh heart, oesophagus and thigh muscle samples from 50 Yorkshire and Landrace pigs were collected from two local abattoirs in Perak from May to August 2014. All the fresh muscle samples were thoroughly examined for macrocyst-forming Sarcocystis spp. and processed using the peptic digestion technique to detect bradyzoites. The results from the muscle samples showed that 58% (29 out of 50) of the pigs were positive for Sarcocystis spp. These findings highlight the importance of implementing stringent measures for screening pigs in abattoirs for Sarcocystis spp. infection because this infection in pigs is a public health concern. PMID:28228924

  16. Experimental Infection of Sarcocystis suihominis in Pig and Human Volunteer in Guangxi%广西猪人肉孢子虫实验感染研究

    Institute of Scientific and Technical Information of China (English)

    李锦辉; 林珍; 杜进发; 覃业新

    2007-01-01

    目的 研究确定广西是否存在人-猪相互感染的肉孢子虫病.方法 采用人-猪-人循环感染方法.取自然感染病例的粪便,用薛氏糖漂浮法收集肉孢子虫孢子囊(10000个以上),拌入饲料,用吞食法感染猪.取含成熟肉孢子虫包囊的新鲜生猪肉,剁碎,用吞食法感染志愿者(含包囊数约71 000个以上).观察感染后症状及肉孢子虫在宿主体内发育情况.结果 人感染后约5 h出现腹胀,8~36 h腹泻水样便13次,呕吐4次,畏寒、发热(体温达38.5℃)、头昏、头痛、关节及肌肉酸痛、腰痛、上腹部隐痛,腿部抽筋,全身乏力,食欲明显减退.感染后第10天粪检查见未孢子化的孢子囊,第12天检出孢子化的孢子囊,孢子囊平均大小为11.9(8.8~14.5) μm×9.2(7.5~12.5) μm.猪感染后第5~8 天轻度厌食,疲乏无力、便秘、毛疏松,第17天恢复正常.包囊平均大小为299.2(175~575) μm×62.3(30~102.5) μm.缓殖子大小为11.5(9.5~13.5) μm×4.1(2.8~5.0) μm.人感染后第46天口服乙酰螺旋霉素(0.2 g/次,4次/d,连服15 d),服药后第30天粪检孢子囊阴转.结论 广西存在猪-人相互感染的肉孢子虫病.

  17. Prevalence of antibodies to Trypanosoma cruzi, Leishmania infantum, Encephalitozoon cuniculi, Sarcocystis neurona, and Neospora caninum in capybara, Hydrochoerus hydrochaeris, from São Paulo State, Brazil

    OpenAIRE

    2010-01-01

    Little is known about the importance of capybara. Hydrochoerus hydrochaeris, as reservoirs for parasites of zoonotic or veterinary importance. Sera from 63 capybaras, from 6 counties in the state of Sao Paulo, Brazil, were examined for antibodies to Trypanosoma cruel, Leishmania infantum, Encephalitozoon cuniculi. Sarcacystis neurona, and Neospora caninum using an indirect immunofluorescent antibody test. Five (8%) of the 63 capybaras had antibodies to T cruzi epimastigotes. None of the sampl...

  18. Meningoencephalitis associated with disseminated sarcocystosis in a free-ranging moose (Alces alces) calf.

    Science.gov (United States)

    Ravi, Madhu; Patel, Jagdish; Pybus, Margo; Coleman, James K; Childress, April L; Wellehan, James F X

    2015-08-01

    A wild moose (Alces alces) calf was presented for necropsy due to severe neurologic signs. Histopathologic examination revealed multisystemic inflammation with intralesional mature and immature schizonts. Schizonts in the brain reacted positively to Sarcocystis spp. polyclonal antibodies. Gene sequencing of PCR-amplified DNA identified the species as Sarcocystis alceslatrans.

  19. Evidence for the presence of an autoimmune component to the chronic muscle wasting disease characteristic of calves infected with Aarcocystis cruzi

    Science.gov (United States)

    Infection with Sarcocystis spp. often resolves in a progressive decline in muscle integrity. The underlying cause for this has remained undetermined. Previously, we described the presence of proinflammatory muscle protein nitration (PMPN) in calves (ScI) chronically infected with Sarcocystis cruzi. ...

  20. Mieloencefalite protozoária eqüina (Relato de caso.

    Directory of Open Access Journals (Sweden)

    R. C. Gonçalves

    2005-03-01

    Full Text Available RESUMO :O objetivo deste trabalho foi relatar um caso de mieloencefalite protozoária eqüina no Estado de São Paulo, Brasil. O diagnóstico se baseou nos sinais clínicos, no resultado positivo para anticorpos contra Sarcocystis neurona no soro e no líquor pela técnica de Western blot. Palavras chave: Mieloencefalite, Sarcocystis neurona, eqüinos. SUMMARY: The purpose of this work was to present a case of equine protozoal myeloencephalitis (EPM in the State of São Paulo, Brazil. The diagnosis was based on the clinical sings, the positive results of the serum and cerebrospinal fluid analysis (Western blot for antibodies against Sarcocystis neurona. Keywords: Myeloencephalitis, Sarcocystis neurona,

  1. Zoonotic parasites from exotic meat in Malaysia.

    Science.gov (United States)

    Fazly, Z A; Nurulaini, R; Shafarin, M S; Fariza, N J; Zawida, Z; Muhamad, H Y; Adnan, M; Premaalatha, B; Erwanas, A I; Zaini, C M; Ong, C C; Chandrawathani, P

    2013-09-01

    Four zoonotic parasites, Sarcocystis spp., Toxoplasma gondii, Trichinella spp. and Taenia spp were screened in exotic meats. A total of forty-six (n=46) meat samples from various species of exotic animals were received from all the 14 states in Malaysia from January 2012 to April 2012. All exotic meat samples were examined macroscopically and histologically for the four zoonotic parasites. Results by histological examination of exotic meats showed the presence of Sarcocystis and Toxoplasma cysts at 8.7% (n=4) and 4.3% (n=2) respectively. No Trichinella spp. and Taenia spp. were found.

  2. An outbreak of sarcocystosis in psittacines and a pigeon in a zoological collection in Brazil.

    Science.gov (United States)

    Ecco, R; Luppi, M M; Malta, M C C; Araújo, M R; Guedes, R M C; Shivaprasad, H L

    2008-12-01

    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.

  3. Developmental biology of Cystoisospora (Apicomplexa: Sarcocystidae) monozoic tissue cysts

    Science.gov (United States)

    Tissue cyst stages are an intriguing aspect of the developmental cycle and transmission of members of the Family Sarcocystidae. Tissue cyst stages of the genera Toxoplasma, Hammondia, Neospora, Besnoitia, and Sarcocystis contain many infectious stages (bradyzoites).The tissue cyst stage of Cystoisos...

  4. Utilización del patrón de restricción del DNA codificante para el RNA Ribosomal de la subunidad pequeña para la caracterización de Apicomplexa

    Directory of Open Access Journals (Sweden)

    López Adelaida

    1996-12-01

    Full Text Available Los Apicomplexos constituyen un phylum de protozoarios que se caracterizan por ser parásitos obligados de una gran variedad de huéspedes vertebrados e invertebrados. Hoy en día hay fuertes polémicas en tomo a su clasificación taxonómica, sus relaciones filogenéticas, y los patrones de coevolución con sus hospederos. El gen que codifica para el ARN ribosomal de la subunidad pequeña (ARN-SURp se utiliza como marcador molecular para resolver estas inquietudes. A partir del ADN de las especies de la familia Sarcocystidae (Sarcocystis cruzi, Sarcocystis sp. de Didelphis marsupialis y Sarcocystis sp. de Columbina talpacoti y Toxoplasma gondii, y de especies de la familia Plasmodiidae (Plasmodium de Anolis chloris, P. simium, y P. falciparumi, se amplificó por PCR el gen que codifica para el ARN de la subunidad ribosomal pequeña (ARN- SURp usando los iniciadores P5-P3, 0009-2134 Y566R-567R. Se compararon
    los patrones de restricción Hind III, Eco RI, Sau 3AI y Alw 261 del DNA ribosomal. La prueba de riboprini mostró que además de discriminar entre familias permite caracterizar diferencias a nivel de género y especie.Apicomplexa is a Protozoa phylum in which all members are obliged parasites of a wide range of vertebrate and invertebrate hosts. There is an ongoing controversy on
    systematics, phylogenetic relationships and parasite - host coevolution patterns. The SSU ribosomal gen has been used as a molecular marker in order to solve these issues.
    From DNA of the species of the Sarcocystidae family (Sarcocystis cruzi, Sarcocystis sp. from Didelphis marsupialis, Sarcocystis sp. from Columbina talpacoti and Toxoplasma
    gondii, and from the species of the Plasmodiidae family (Plasmodium from Anolis ehloris, P. simium, and P. falciparum, the SSU ribosomal DNA fragemnt was
    amplified by PCR, using the pair of primers P5-P3, 0009-2134 and 566R-567R. Hind III, Eco RI, Sau 3AI and Alw 261 restriction pattems were compared

  5. PARASITES TRANSMITTED TO HUMAN BY INGESTION OF DIFFERENT TYPES OF MEAT, EL-MINIA CITY, EL-MINIA GOVERNORATE, EGYPT.

    Science.gov (United States)

    Abdel-Hafeez, Ekhlas Hamed; Kamal, Amany Mohamed; Abdelgelil, Noha Hamed; Abdel-Fatah, Mohamed

    2015-12-01

    Meat-borne parasites are Sarcocystis species, Toxoplasma gondii, Taenia saginata, Taenia solium and Trichinella spiralis. A total of 300 animals including 100 cattle, 100 goat, and 100 pigs, slaughtered in El-Minia governmental slaughterhouses. From each animal, five samples were taken from different muscles (esophageal, tongue and cardiac) and different organs (liver and brain). Meat samples were examined macroscopic and microscopic (direct, homogenization and H&E staining) for detection of the above-mentioned parasites. Serum samples were subjected to IHA for detection of T gondii specific antibodies. This study revealed that Sarcocystis species were the highest parasites that could be detected, with overall prevalence of 80%, which was statistically significant (P pigs were infected with C. cellulosae, but without statistical significant (P < or = 0.5).

  6. HISTOPATHOLOGICAL SURVEY OF PROTOZOA, HELMINTHS AND ACARIDS OF IMPORTED AND LOCAL PSITTACINE AND PASSERINE BIRDS IN JAPAN

    OpenAIRE

    Tsai, Shinn-Shyong; HIRAI, Katsuya; ITAKURA, Chitoshi

    1992-01-01

    A total of 534 psittacine and passerine birds consisting of 241 imported and 293 local birds were examined histologically. As a result, the following parasites were found : Giardia (86 cases), Knemido-coptes (26 cases), coccidia (10 cases), Ascaridia (6 cases), Cryptosporidium (5 cases), Sarcocystis (5 cases), tapeworm (4 cases), microfilaria (2 cases), Hexamita (1 case), and Spiroptera (1 case). High incidences of giardiasis and knemido-coptic infestation were detected in the local birds, bu...

  7. Suspected new wave of muscular sarcocystosis in travellers returning from Tioman Island, Malaysia, May 2014.

    Science.gov (United States)

    Tappe, D; Stich, A; Langeheinecke, A; von Sonnenburg, F; Muntau, B; Schäfer, J; Slesak, G

    2014-05-29

    In May 2014, six patients presented in Germany with a Sarcocystis-associated febrile myositis syndrome after returning from Tioman Island, Malaysia. During two earlier waves of infections, in 2011 and 2012, about 100 travellers returning to various European countries from the island were affected. While the first two waves were associated with travel to Tioman Island mostly during the summer months, this current series of infections is associated with travel in early spring, possibly indicating an upcoming new epidemic.

  8. Human Invasive Muscular Sarcocystosis Induces Th2 Cytokine Polarization and Biphasic Cytokine Changes, Based on an Investigation among Travelers Returning from Tioman Island, Malaysia.

    Science.gov (United States)

    Tappe, Dennis; Slesak, Günther; Pérez-Girón, José Vicente; Schäfer, Johannes; Langeheinecke, Andreas; Just-Nübling, Gudrun; Muñoz-Fontela, César; Püllmann, Kerstin

    2015-06-01

    Sarcocystis nesbitti is a parasite responsible for a biphasic eosinophilic febrile myositis syndrome in two recent outbreaks in Malaysia. We demonstrate Th2 cytokine polarization in infected travelers, an overall cytokine production decrease in the early phase of the disease suggestive of initial immunosuppression, and elevated levels of proinflammatory and chemotactic cytokines in the later myositic phase. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  9. Sarcocystinae: nomina dubia and available names.

    Science.gov (United States)

    Frenkel, J K; Heydorn, A O; Mehlhorn, H; Rommel, M

    1979-02-28

    Examination of the original descriptions of the species of Sarcocystis in cattle, sheep, and swine, and of isosporid oocysts shed sporulated by dogs, cats, man, and other carnivores, has shown that it is not possible in most instances to identify unambiguously recently recognized taxa. The original descriptions are insufficient, and because no type specimens exist, could apply to two or more of the presently recognized taxa. We consider the following nomina dubia: Sarcocystis hirsuta S. miescheriana S. tenella S. cruzi S. bertrami Isospora bigemina (S. bigemina) I. hominis (S. hominis) I. buteonis (Frenkelia buteonis) Because the former type species, Sarcocystis miescheriana, is an indeterminate nomen dubium, we are proposing S. muris as the new type species. Historically, it was the first species described clearly and unambiguously even in the light of present knowledge, and the stages of its life cycle are probably completely known; it was the second species to be named. Old and recent descriptions are reviewed, and definitions are proposed for the following taxa: S. bovifelis S. bovicanis S. bovihominis S. ovifelis S. ovicanis S. muris (type species) S. suihominis S. suicanis S. equicanis Frenkelia microti F. glareoli for which neotypes will be prepared and deposited with designated institutions and curators. A new subfamily, Cystoisosporinae, is created.

  10. Description of two new Isospora species causing visceral coccidiosis in captive superb glossy starlings, Lamprotornis superbus (Aves: Sturnidae).

    Science.gov (United States)

    Hafeez, Mian A; Stasiak, Iga; Delnatte, Pauline; El-Sherry, Shiem; Smith, Dale A; Barta, John R

    2014-09-01

    Isospora greineri sp. n. and Isospora superbusi sp. n. are described from captive superb glossy starlings, Lamprotornis superbus, from the Toronto Zoo succumbing to visceral coccidiosis. Sequence data from nuclear 18S recombinant DNA (rDNA) and mitochondrial cytochrome c oxidase subunit I (COI) loci from sporulated oocysts and infected tissues (liver, lung, or spleen) demonstrated two distinct Isospora sp. genotypes that varied in their relative abundance. In the tissues of one affected bird, as well as its associated fecal sample, two distinct COI sequences (1.7% divergence) and two distinct 18S rDNA sequences (0.6% divergence) were found at almost the same abundance; in other specimens, one of the 18S and one of the COI sequences were less abundant than the other. In the tissues of some birds, only a single COI and single 18S sequence were present. In all cases, the same pair of 18S rDNA and COI sequences fluctuated in abundance in parallel, indicating that there were two distinct species present rather than one species with more than one COI or 18S locus. The oocysts of these new species cannot be differentiated morphologically. Sporulated oocysts of both were spherical to subspherical measuring 17.7 ± 0.22 μm by 17.1 ± 0.20 μm with a mean L/W ratio of 1.03 ± 0.004. Sporocysts were ovoid measuring 13.5 ± 0.17 μm by 9.3 ± 0.15 μm with a mean L/W ratio of 1.4 ± 0.02. Sporocysts had a small Stieda body with indistinct sub-Stieda body; each sporocyst had a compact residuum. Two morphologically similar but genetically divergent Isospora species were shown to cause simultaneous enteric and extraintestinal infections in captive superb glossy starlings.

  11. Parasites, diseases, and health status of sympatric populations of sika deer and white-tailed deer in Maryland and Virginia.

    Science.gov (United States)

    Davidson, W R; Crow, C B

    1983-10-01

    In July 1981, investigations on parasites, diseases, and herd health status were conducted on sympatric populations of sika deer (Cervus nippon) and white-tailed deer (Odocoileus virginianus) from Blackwater National Wildlife Refuge (Maryland) and Chincoteague National Wildlife Refuge (Virginia) on the Delmarva Peninsula. Five adult deer of each species were collected from each location and subjected to thorough necropsy examinations and laboratory tests. White-tailed deer at both locations harbored protozoan, helminth, and arthropod parasites typically associated with this species throughout the southeastern United States. In contrast, sika deer at both locations harbored only light burdens of ticks, chiggers, and sarcocysts. Serologic tests for antibodies to seven infectious disease agents revealed evidence of exposure to bovine virus diarrhea (BVD) virus, infectious bovine rhinotracheitis virus, and parainfluenza3 virus in white-tailed deer, but only BVD virus in sika deer. At both locations the general health status of sika deer was superior to that of white-tailed deer.

  12. Immunohistochemical diagnosis of Neospora caninum in tissue sections.

    Science.gov (United States)

    Lindsay, D S; Dubey, J P

    1989-11-01

    An avidin-biotin-peroxidase complex immunoperoxidase staining method was developed to detect Neospora caninum in formalin-fixed, paraffin-embedded tissue sections. Specific antiserum to N caninum was made in rabbits and used to probe tissues from dogs naturally and experimentally infected with N caninum. The test detected tachyzoites and bradyzoites of N caninum. A reaction was not observed to Toxoplasma gondii, Hammondia hammondi, Sarcocystis cruzi, S capricanis, S tenella, Besnoitia jellisoni, Caryospora bigenetica, Hepatazoon canis, Atoxoplasma sp, or the organism causing canine dermal coccidiosis. When antiserum made in rabbits to T gondii was used in the test, reaction to N caninum was not observed.

  13. Red foxes (Vulpes vulpes) in Ireland as hosts for parasites of potential zoonotic and veterinary significance.

    Science.gov (United States)

    Wolfe, A; Hogan, S; Maguire, D; Fitzpatrick, C; Vaughan, L; Wall, D; Hayden, T J; Mulcahy, G

    Intestinal washes, faecal flotations and serological examinations for antibodies to Toxoplasma gondii and Neospora caninum were used to assess the prevalence of parasites in carcases of foxes killed on roads or shot in the Dublin area and surrounding counties. The ascarids Uncinaria stenocephala and Toxocara canis were prevalent, as was the trematode Alaria alata. Taenia species, eggs of Capillaria species and sporocysts of Sarcocystis species were also found. Only one fox out of 70 examined was seropositive for N. caninum, whereas 24 of 51 were seropositive for T. gondii.

  14. Studies on endoparasites of the black bear (Ursus americanus) in the southeastern United States.

    Science.gov (United States)

    Crum, J M; Nettles, V F; Davidson, W R

    1978-04-01

    Examination of 53 black bears (Ursus americanus) from six states in the southeastern United States revealed at least 17 species of endoparasites, including Sarcocystis sp., Spirometra mansonoides (spargana), Macracanthorhynchus ingens, Ancylostoma caninum, Arthrocephalus lotoris, Baylisascaris transfuga, Capillaria aerophila, Capillaria putorii, Crenosoma sp., Cyathospirura sp., Dirofilaria immitis, Gnathostoma sp., Gongylonema pulchrum, microfilariae, Molineus barbatus, Physaloptera sp. and Strongyloides sp. Twelve of these represent new host records for black bear, and two are considered to be new species. Data are presented on prevalence, intensity and geographic distribution of each species. Pathologic effects were associated with infections of spargana of S. mansonoides and adults of C. aerophilia.

  15. Comparison of three concentration methods for the recovery of canine intestinal parasites from stool samples.

    Science.gov (United States)

    Katagiri, S; Oliveira-Sequeira, T C G

    2010-10-01

    The aim of present study was to compare the efficiency of a commercial assay and two conventional methods for fecal concentration in detecting canine gastrointestinal parasites. Fecal samples from 254 dogs were processed by centrifugation-sedimentation (CS), centrifugation-flotation (CF) and a commercial assay for fecal concentration (TF-test). The following parasites were detected: Ancylostoma (37.8%), Giardia (16.9%), Toxocara canis (8.7%), Trichuris vulpis (7.1%), Isospora (3.5%), and Sarcocystis (2.7%). The calculated analytical sensitivity indicated that CF was more accurate (Pindividual diagnosis and epidemiological investigation.

  16. Alguns ectoparasitas e protozoários em bovinos da República da Guiné-Bissau

    OpenAIRE

    ROSA, FERNANDA; Crespo, Maria Virgínia; Travassos Dias, J. A.

    1998-01-01

    Os autores assinalam a presença de algumas espécies de ixodídeos (Amblyomma variegatum, Boophilus annulatus, B. geigy e Hyalomma sp.) localizados em diferentes regiões do corpo de bovinos autóctones da República da Guiné-Bissau, colhidos entre Novembro de 1990 e Março de 1991. A pesquisa de protozoários, efectuada principalmente em esfregaços de sangue revelou a presença de Anaplasma marginale, Theileria mutans, Babesia bigemina e Sarcocystis sp. Não foram encontradas lesões relacionada...

  17. Developmental biology of Cystoisospora (Apicomplexa: Sarcocystidae) monozoic tissue cysts.

    Science.gov (United States)

    Lindsay, David S; Houk, Alice E; Mitchell, Sheila M; Dubey, J P

    2014-08-01

    Tissue cyst stages are an intriguing aspect of the developmental cycle and transmission of species of Sarcocystidae. Tissue-cyst stages of Toxoplasma, Hammondia, Neospora, Besnoitia, and Sarcocystis contain many infectious stages (bradyzoites). The tissue cyst stage of Cystoisospora (syn. Isospora) possesses only 1 infectious stage (zoite), and is therefore referred to as a monozoic tissue cyst (MZTC). No tissue cyst stages are presently known for members of Nephroisospora. The present report examines the developmental biology of MZTC stages of Cystoisospora Frenkel, 1977 . These parasites cause intestinal coccidiosis in cats, dogs, pigs, and humans. The MZTC stages of C. belli are believed to be associated with reoccurrence of clinical disease in humans.

  18. [Emerging parasitic diseases].

    Science.gov (United States)

    Weibel Galluzzo, C; Wagner, N; Michel, Y; Jackson, Y; Chappuis, F

    2014-05-07

    Travels, migration and circulation of goods facilitate the emergence of new infectious diseases often unrecognized outside endemic areas. Most of emerging infections are of viral origin. Muscular Sarcocystis infection, an acute illness acquired during short trips to Malaysia, and Chagas disease, a chronic illness with long incubation period found among Latin American migrants, are two very different examples of emerging parasitic diseases. The former requires a preventive approach for travelers going to Malaysia and must be brought forth when they return with fever, myalgia and eosinophilia, while the latter requires a proactive attitude to screen Latin American migrant populations that may face difficulties in accessing care.

  19. Some diseases and parasites of captive woodcocks

    Science.gov (United States)

    Locke, L.N.; Stickel, W.H.; Geis, S.A.

    1965-01-01

    Observations were made concerning the diseases and parasites of a group of woodcocks (Philohela minor) caught in Massachusetts in the summer of 1960 and kept in captivity in Maryland, and of another group caught and kept in Louisiana in the winter of 1960-61. Bumblefoot, a granulomatous swelling of the foot caused by Micrococcus sp., is reported for woodcocks for the first time. Six of 31 woodcocks were infected with a renal coccidium of an undetermined species. Tetrameres sp. was found in 4 of 31 birds examined. Sarcocystis was found in one bird. Aerosaculitis was found in several.

  20. Molecular phylogeny of Toxoplasmatinae: comparison between inferences based on mitochondrial and apicoplast genetic sequences

    Directory of Open Access Journals (Sweden)

    Michelle Klein Sercundes

    2016-03-01

    Full Text Available Abstract Phylogenies within Toxoplasmatinae have been widely investigated with different molecular markers. Here, we studied molecular phylogenies of the Toxoplasmatinae subfamily based on apicoplast and mitochondrial genes. Partial sequences of apicoplast genes coding for caseinolytic protease (clpC and beta subunit of RNA polymerase (rpoB, and mitochondrial gene coding for cytochrome B (cytB were analyzed. Laboratory-adapted strains of the closely related parasites Sarcocystis falcatula and Sarcocystis neurona were investigated, along with Neospora caninum, Neospora hughesi, Toxoplasma gondii (strains RH, CTG and PTG, Besnoitia akodoni, Hammondia hammondiand two genetically divergent lineages of Hammondia heydorni. The molecular analysis based on organellar genes did not clearly differentiate between N. caninum and N. hughesi, but the two lineages of H. heydorni were confirmed. Slight differences between the strains of S. falcatula and S. neurona were encountered in all markers. In conclusion, congruent phylogenies were inferred from the three different genes and they might be used for screening undescribed sarcocystid parasites in order to ascertain their phylogenetic relationships with organisms of the family Sarcocystidae. The evolutionary studies based on organelar genes confirm that the genusHammondia is paraphyletic. The primers used for amplification of clpC and rpoB were able to amplify genetic sequences of organisms of the genus Sarcocystisand organisms of the subfamily Toxoplasmatinae as well.

  1. Coccidian parasites (Apicomplexa) from snakes in the southcentral and southwestern United States: new host and geographic records.

    Science.gov (United States)

    McAllister, C T; Upton, S J; Trauth, S E; Dixon, J R

    1995-02-01

    Four hundred thirty-five leptotyphlopid, colubrid, elapid, and viperid snakes were collected from various localities in Arkansas, New Mexico, Oklahoma, and Texas, and their feces were examined for coccidian parasites. Of these, 131 (30%) were passing oocysts or sporocysts of at least 1 coccidian; 88 (67%) of the infected snakes had only 1 species of coccidian when they were examined. Aquatic and semiaquatic snakes accounted for 48% of the infections, whereas strictly terrestrial snakes comprised the other 52%. There was more than a 2-fold difference in prevalence among these 2 groups as 63 of 129 (49%) of the aquatic and semiaquatic snakes versus 68 of 306 (22%) of the terrestrial snakes harbored coccidia. Most terrestrial snakes were infected by species of Caryospora and Sarcocystis that are either facultatively or obligatorily heteroxenous. The aquatic and semiaquatic species most often harbored eimerians. Attempts to transmit some of the Sarcocystis spp. experimentally from Crotalus atrox to Mus musculus, Peromyscus leucopus, Peromyscus maniculatus, or Microtus ochrogaster were unsuccessful. This report documents 27 new host and several distributional records for coccidians from snakes in the southcentral and southwestern United States.

  2. Hepatic sarcocystosis in a striped dolphin (Stenella coeruleoalba) from the Spanish Mediterranean coast.

    Science.gov (United States)

    Resendes, A R; Juan-Sallés, C; Almeria, S; Majó, N; Domingo, M; Dubey, J P

    2002-02-01

    Fatal hepatic sarcocystosis was diagnosed in a striped dolphin (Stenella coeruleoalba) from the northeastern Spanish Mediterranean coast based on pathologic findings and the microscopic and ultrastructural characteristics of the intralesional parasite. Main gross lesions were icterus, subcutaneous hemorrhages, and hepatic congestion. The most prominent microscopic lesions consisted of severe acute multifocal to coalescing necrotizing hepatitis with cholestasis and intralesional protozoa. There was severe chronic pancreatitis with generalized distension of pancreatic ducts by hyaline plugs and adult trematodes. Only asexual stages of the protozoa were found. The parasite in the liver divided by endopolygeny. Schizonts varied in shape and size. Mature schizonts had merozoites randomly arranged or budding peripherally around a central residual body. Schizonts were up to 22 microm long, and merozoites were up to 6 microm long. Ultrastructurally, merozoites lacked rhoptries. This parasite failed to react by immunohistochemistry with anti-Toxoplasma gondii, anti-Neospora caninum and anti-Sarcocystis neurona antibodies. The microscopic and ultrastructural morphologies of the parasite were consistent with Sarcocystis canis, so far described only from animals in the Unites States. The life cycle and source of S. canis are unknown. The present report of S. canis-like infection in a sea mammal from Spain indicates that the definitive host for this parasite also exists outside of the United States.

  3. An epidemiological study of gastrointestinal parasites of dogs from Southern Greater Buenos Aires (Argentina): age, gender, breed, mixed infections, and seasonal and spatial patterns.

    Science.gov (United States)

    Fontanarrosa, María F; Vezzani, Darío; Basabe, Julia; Eiras, Diego F

    2006-03-31

    A total of 2193 fecal samples from owned dogs were collected during the 2003-2004 period in Southern Greater Buenos Aires, and were evaluated for the presence of intestinal parasites by a flotation-centrifugation method. The overall prevalence was 52.4%, and the 11 species found were: Ancylostoma caninum (13%), Isospora ohioensis complex (12%), Toxocara canis (11%), Trichuris vulpis (10%), Sarcocystis sp. (10%), Giardia duodenalis (9%), Isospora canis (3%), Hammondia-Neospora complex (3%), Dipilydium caninum (18 cases), Cryptosporidium sp. (5 cases), and Toxascaris leonina (1 case). There was no significant difference in the overall prevalence between genders (female = 50.4%, male = 54.6%), and breeds (pure = 52.3%, mixed = 53%), but prevalence in puppies (canis, A. caninum, and T. vulpis were spatially heterogeneous with a clear Southwest-Northeast gradient. Only prevalences of Sarcocystis sp. and G. duodenalis showed seasonal variation. The frequency distribution of the number of species per fecal sample did not differ from a random distribution. Results obtained throughout the world were discussed.

  4. Fecal shedding of Toxocara canis and other parasites in foxes and coyotes on Prince Edward Island, Canada.

    Science.gov (United States)

    Wapenaar, Wendela; Barkema, Herman W; O'Handley, Ryan

    2013-04-01

    Knowledge of parasites shed by wild canids can assist in recognizing risk to human and domestic animal health. Our aim was to estimate the prevalence of patent infections with Toxocara canis and other parasites in foxes (Vulpes vulpes) and coyotes (Canis latrans) in Prince Edward Island, Canada. Identification of parasite species was based on microscopic examination of feces, with the use of a sucrose fecal flotation method. Sample collection was performed in winter on carcasses of 271 and 185 hunted or trapped foxes and coyotes, respectively. One or more parasite species were observed in 242 (89%) foxes and 128 (69%) coyotes. Toxocara canis, Uncinaria stenocephala, Capillaria spp., Mesocestoides, Taenidd spp., Alaria spp., Cryptocotyle lingua, Sarcocystis spp., Neospora caninum-like coccidia, and other coccidia were identified. A third of juvenile foxes were shedding T. canis and had a high prevalence of Capillaria spp., especially in juvenile foxes (69%). Taenidd eggs, Alaria spp. and Sarcocystis spp. were more common in coyotes (24, 18, and 9%, respectively) than foxes (8, 11, and 1%, respectively). Despite the limitations of fecal flotation to identify parasite species, the high prevalence of T. canis warrants the attention of public health professionals.

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

    Science.gov (United States)

    Allison, Robin; Williams, Philip; Lansdowne, Jennifer; Lappin, Michael; Jensen, Tracey; Lindsay, David

    2006-09-01

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

  6. Histopathological survey of protozoa, helminths and acarids of imported and local psittacine and passerine birds in Japan.

    Science.gov (United States)

    Tsai, S S; Hirai, K; Itakura, C

    1992-12-01

    A total of 534 psittacine and passerine birds consisting of 241 imported and 293 local birds were examined histologically. As a result, the following parasites were found: Giardia (86 cases), Knemido-coptes (26 cases), coccidia (10 cases), Ascaridia (6 cases), Cryptosporidium (5 cases), Sarcocystis (5 cases), tapeworm (4 cases), microfilaria (2 cases), Hexamita (1 case), and Spiroptera (1 case). High incidences of giardiasis and knemido-coptic infestation were detected in the local birds, but rarely in the imported birds. Giardial trophozoites were observed mainly in the duodenum of budgerigars (Melopsittacus undulatus). Knemidocoptic mites burrowed into the epidermis producing proliferative dermatitis in 25 budgerigars and 1 African Grey Parrot (Psittacus erithacus erithacus). This ectoparasite often infested the skin around the cloaca. Coccidiosis was seen only in the small intestines of the finch (Poephila gouldiae gouldiae), African Grey Parrot, Rainbow lory (Trichoglossus haematodus), Indian Ring-necked parakeet (Psittacula krameri manillensis) and peach-faced lovebird (Agapornis roseicollis). Two parrots (Amazona aestiva aestiva and Psittacus erithacus erithacus) and two budgerigars had intestinal cryptosporidiosis. Conjunctivitis associated with cryptosporidial infection was seen in a lovebird. Sarcocystis cysts containing crescent-shaped bradyzoites were found not only in the thigh and breast but also in the heart and cloacal muscles. Other organisms such as Ascaridia, tapeworm, microfilaria, Hexamita, and Spiroptera were clinically less significant. However, infections such as Giardia and Cryptosporidim might have zoonotic implications.

  7. Epidemiology and control prospects of foodborne parasitic zoonoses in the European Union.

    Science.gov (United States)

    Pozio, E

    2008-06-01

    In the 27 Member States of the European Union, zoonotic parasites transmitted by food are circulating with different prevalence according to the country, the environmental conditions, the human behaviour, and the socio-economic level. Foodborne parasites can be divided in two main groups according to the way of transmission to humans. These foodborne parasites reach the human beings through the consumption of raw infected food such as muscle tissues of different animal species (Toxoplasma gondii, Sarcocystis hominis, Sarcocystis suishominis, Diphyllobotrium latum, Taenia solium, Taenia saginata, Opisthorchis felineus, Anisakis spp., Pseudoterranova spp., Trichinella spp.), or vegetables (Fasciola hepatica), and contaminated food and water resources (Giardia duodenalis, Cryptosporidium spp., T. gondii, Echinococcus granulosus sensu latu, Echinococcus multilocularis, T. solium, Taenia multiceps). As a general role, the control strategies should be based on the education of the consumers, farmers and shepherds, the improvement of farming conditions, the improvement or the development of more sensitive methods to detect these parasites in slaughtered animals and in foodstuff, a control of sewage sludge on pastures and of drinking water resources, and the reduction of contacts between livestock and wild animals which frequently represent the most important reservoir of these pathogens.

  8. Survey of Feral Swine ( Sus scrofa ) Infection with the Agent of Chagas Disease ( Trypanosoma cruzi ) in Texas, 2013-14.

    Science.gov (United States)

    Comeaux, Juliette M; Curtis-Robles, Rachel; Lewis, Barbara C; Cummings, Kevin J; Mesenbrink, Brian T; Leland, Bruce R; Bodenchuk, Michael J; Hamer, Sarah A

    2016-07-01

    : Feral swine ( Sus scrofa ) are an invasive species and reservoir of numerous zoonotic pathogens in the US, and Texas leads the nation in the estimated population size of feral hogs. Texas also harbors enzootic transmission cycles of the protozoan parasite Trypanosoma cruzi , agent of Chagas disease. Given previous evidence that swine can serve as reservoirs of T. cruzi in Latin America and new evidence of triatomines (kissing bugs) feeding on swine in Texas, we measured the prevalence of T. cruzi infection in feral swine in Texas. From 2013 to 2014, we sampled blood and/or cardiac tissue from 78 feral swine across 14 Texas counties (seven with and seven without prior documentation of kissing bug occurrence) and used PCR and histopathology to detect T. cruzi infection. We determined an overall infection prevalence of 6% (3 of 54) based on PCR evaluation of cardiac tissue, and no blood samples were positive (n=72). All three positive pigs were from counties where kissing bugs are documented. No T. cruzi amastigotes were noted on histopathology (n=54). Sarcocysts were observed in 10 (18%) of the samples, five of which also had mild focal areas of degeneration and inflammatory cell infiltration. Eco-epidemiologic investigations can provide an assessment of contributions of feral hogs to maintenance of T. cruzi across a landscape to help protect human and animal health.

  9. Endoparasites of American marten (Martes americana): Review of the literature and parasite survey of reintroduced American marten in Michigan.

    Science.gov (United States)

    Spriggs, Maria C; Kaloustian, Lisa L; Gerhold, Richard W

    2016-12-01

    The American marten (Martes americana) was reintroduced to both the Upper (UP) and northern Lower Peninsula (NLP) of Michigan during the 20th century. This is the first report of endoparasites of American marten from the NLP. Faeces from live-trapped American marten were examined for the presence of parasitic ova, and blood samples were obtained for haematocrit evaluation. The most prevalent parasites were Capillaria and Alaria species. Helminth parasites reported in American marten for the first time include Eucoleus boehmi, hookworm, and Hymenolepis and Strongyloides species. This is the first report of shedding of Sarcocystis species sporocysts in an American marten and identification of 2 coccidian parasites, Cystoisospora and Eimeria species. The pathologic and zoonotic potential of each parasite species is discussed, and previous reports of endoparasites of the American marten in North America are reviewed.

  10. SARCOSPORIDIOSIS - MEDICAL IMPORTANCE AND DIAGNOSIS

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    Milena Misic

    2004-07-01

    Full Text Available Sarcosporidiosis (Sarcocystis infection is caused by an intracellular protozoan parasite that predominantly affects animals. It can rarely be found in human skeletal and cardiac muscle in humans. There are two different forms of sarcosporidiosis in humans. These cases of muscular sarcocystosis were probably zoonotic in origin and associated with close contact with definitive hosts (both domestic and wild animals thus permitting the contamination of food and drink with sporocystis shed by these definitive hosts. The second mode of infection for humans is ingested animal tissues which containing sporozoites (e.g., undercooked meats. These sporozoited directly intestinal epithelial cells and can enter the circulation in an manner similiar to those released from oocysts from the intermediate or accidental host.

  11. Endoparasites of American marten (Martes americana: Review of the literature and parasite survey of reintroduced American marten in Michigan

    Directory of Open Access Journals (Sweden)

    Maria C. Spriggs

    2016-12-01

    Full Text Available The American marten (Martes americana was reintroduced to both the Upper (UP and northern Lower Peninsula (NLP of Michigan during the 20th century. This is the first report of endoparasites of American marten from the NLP. Faeces from live-trapped American marten were examined for the presence of parasitic ova, and blood samples were obtained for haematocrit evaluation. The most prevalent parasites were Capillaria and Alaria species. Helminth parasites reported in American marten for the first time include Eucoleus boehmi, hookworm, and Hymenolepis and Strongyloides species. This is the first report of shedding of Sarcocystis species sporocysts in an American marten and identification of 2 coccidian parasites, Cystoisospora and Eimeria species. The pathologic and zoonotic potential of each parasite species is discussed, and previous reports of endoparasites of the American marten in North America are reviewed.

  12. Frequency of parasites and Salmonella infection in captive maned-wolf, Chrysocyon brachyurus, kept in Zoos at the State of São Paulo, Brazil

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    Gilioli R.

    2000-01-01

    Full Text Available Thirty-one captive maned wolves (Chrysocyon brachyurus, Illiger 1815 from 11 Zoos at the State of São Paulo, Brazil, were screened to investigate the presence of parasites and Salmonella infection by parasitological diagnostic methods and fecal selective culture. The most frequent ecto and endoparasites found were Ctenocephalides felis (56.2%, Rhipicephalus sanguineus (12.5%, Ancylostoma caninum (45.1%, Strongyloides sp. (29.0%, Uncinaria stenocephala (3.2%, Capillaria sp. (3.2%, Entamoeba sp. (22.9%, Sarcocystis sp. (29.0%, Cryptosporidium sp. (19.3%, Eimeria sp. (19.3%, Giardia sp. (9.6% and Isospora sp. (3.2%. Four different serotypes of Salmonella were identified in six animals (25%. Only one infected animal showed clinical signs of diarrhea. The ability to harbor Salmonella spp. as normal nonpathogenic bacteria of the gastrointestinal tract may be a physiological adaptation of this specie.

  13. 青海省海西州天峻县藏羊寄生虫区系调查

    Institute of Scientific and Technical Information of China (English)

    王启菊; 晁生玉; 杨安圈; 倪关英; 卓玛措; 环臧

    2008-01-01

    2007年5月作者对青海省天峻县9个乡镇的29只藏羊进行蠕虫学全身剖检,发现吸虫1种,绦虫2种,绦虫蚴3种,线虫19种,住肉孢子虫1种,蜘蛛昆虫2种.优势虫种为住肉孢子虫(sarcocystis sp)、蒙古马歇尔线虫(marshallagiamongolica)、马氏马歇尔线虫(marshallagia marshalli)、棘球蚴(echinococcus granulosus),感染率分别为84.6%,75.9%、72.4%、62%.

  14. First survey of endoparasites in pet ferrets in Italy.

    Science.gov (United States)

    d'Ovidio, D; Pepe, P; Ianniello, D; Noviello, E; Quinton, Jean-Francois; Cringoli, G; Rinaldi, L

    2014-06-16

    Endoparasites are infrequently reported in ferrets. A cross-sectional survey was conducted to determine the prevalence of intestinal parasites in pet ferrets in southern Italy. Fresh fecal samples were randomly collected from 50 ferrets housed in pet shops or privately owned. All fecal samples were processed using the FLOTAC pellet technique to identify and count helminthic eggs/larvae and protozoan cysts/oocysts. In addition, the samples were analyzed also by the Remel XpectGiardia/Cryptosporidium immunoassay. Intestinal parasites were detected in 15 out of 50 ferrets (30%). Eggs of ancylostomids were found in 28.0% (14/50) of the animals and oocysts of Sarcocystis were detected in one ferret (2.0%). None of the samples was positive for Cryptosporidium or Giardia. To the authors' knowledge, this is the first report of sarcosporidiosis in a pet ferret in Italy.

  15. Chlamydiosis in a red-tailed hawk (Buteo jamaicensis).

    Science.gov (United States)

    Mirandé, L A; Howerth, E W; Poston, R P

    1992-04-01

    A red-tailed hawk (Buteo jamaicensis) with signs of respiratory distress and diarrhea was captured in the Manchac Wildlife Management Area, Louisiana (USA) and died the following day. At necropsy, the carcass was emaciated and there were splenomegaly, and fibrinous pericarditis, airsacculitis, and perihepatitis. Microscopically, there were fibrinous pericarditis and airsacculitis, myocardial necrosis, necrotizing hepatitis, splenic necrosis with reticuloendothelial cell hyperplasia, interstitial pneumonia and focal pancreatic necrosis. Intracytoplasmic chlamydial inclusion bodies were noticed in macrophages in the fibrinous exudate covering air sac and pericardium, and in spleen, liver, heart, lung, and pancreas. Schizonts compatible with a Sarcocystis sp.-like protozoon were present in the walls of air capillaries in the lung. A Chlamydia sp.-like organism was isolated in embryonating chicken eggs and cell culture and identified as C. psittaci with immunofluorescent staining.

  16. Equine Protozoal Myeloencephalitis: An Updated Consensus Statement with a Focus on Parasite Biology, Diagnosis, Treatment, and Prevention.

    Science.gov (United States)

    Reed, S M; Furr, M; Howe, D K; Johnson, A L; MacKay, R J; Morrow, J K; Pusterla, N; Witonsky, S

    2016-01-01

    Equine protozoal myeloencephalitis (EPM) remains an important neurologic disease of horses. There are no pathognomonic clinical signs for the disease. Affected horses can have focal or multifocal central nervous system (CNS) disease. EPM can be difficult to diagnose antemortem. It is caused by either of 2 parasites, Sarcocystis neurona and Neospora hughesi, with much less known about N. hughesi. Although risk factors such as transport stress and breed and age correlations have been identified, biologic factors such as genetic predispositions of individual animals, and parasite-specific factors such as strain differences in virulence, remain largely undetermined. This consensus statement update presents current published knowledge of the parasite biology, host immune response, disease pathogenesis, epidemiology, and risk factors. Importantly, the statement provides recommendations for EPM diagnosis, treatment, and prevention.

  17. Two potentially zoonotic parasites infecting Philippine brown deer (Cervus mariannus desmarest, 1822 in Leyte Island

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    Harvie Potot Portugaliza

    2015-12-01

    Full Text Available This case report describes the necropsy findings of two potentially zoonotic parasites infecting the Philippine brown deer (Cervus mariannus in Leyte Island, Philippines. A female deer aging approximately 5-year was presented for necropsy to the Diagnostic Laboratory at the College of Veterinary Medicine, Visayas State University. Gross pathology was recorded and the selected organs having lesion were collected for histopathological studies. Results showed severe necrotizing lesions in the nasal and palatal areas, infestation of calliphorid maggots, hepatic fibrosis, cholangitis, cholecystitis, lung atelectasis and duodenitis. Heavy ruminal fluke infection was also observed. Two potentially zoonotic parasites namely Fasciola gigantica and Sarcocystis spp. were identified. The Philippine brown deer appears to have a role in transmission and amplification of zoonotic parasites, and can also be threatened by diseases caused by the parasites.

  18. Histopathology of protozoal infection in animals: a retrospective study at the University of Philippines College of Veterinary Medicine (1972-2010).

    Science.gov (United States)

    Baticados, Abigail M; Baticados, Waren N

    2012-01-01

    The authors describe the first parasitological survey of protozoal infections on tissue slide sections of field cases processed at the histopathology laboratory of the College of Veterinary Medicine (CVM) at the University of the Philippines Los Baños (UPLB). Over 80% of the field cases were from Region 4 (CALABARZON) and the rest were equally distributed from other areas of the Philippines, namely: Region 2 (Cagayan Valley), Metropolitan Manila (National Capital Region), Region III (Central Luzon) and Region VI (Western Visayas). Histopathological analyses of tissue sections from 51 archived cases (1972-2010) of parasitic aetiology were performed. Microscopic examination of a total of 286 histopathological slides revealed the presence of several protozoa, including sarcosporidiosis, hepatic coccidiosis, intestinal coccidiosis, balantidiosis and leucocyto-zoonosis. In addition, the finding of Balantidium and Sarcocystis may have zoonotic implications and can therefore be used as markers of public health importance.

  19. Histopathology of protozoal infection in animals: a retrospective study at the University of Philippines College of Veterinary Medicine (1972-2010

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    Abigail M. Baticados

    2012-03-01

    Full Text Available The authors describe the first parasitological survey of protozoal infections on tissue slide sections of field cases processed at the histopathology laboratory of the College of Veterinary Medicine (CVM at the University of the Philippines Los Baños (UPLB. Over 80% of the field cases were from Region 4 (CALABARZON and the rest were equally distributed from other areas of the Philippines, namely: Region 2 (Cagayan Valley, Metropolitan Manila (National Capital Region, Region III (Central Luzon and Region VI (Western Visayas. Histopathological analyses of tissue sections from 51 archived cases (1972-2010 of parasitic aetiology were performed. Microscopic examination of a total of 286 histopathological slides revealed the presence of several protozoa, including sarcosporidiosis, hepatic coccidiosis, intestinal coccidiosis, balantidiosis and leucocyto-zoonosis. In addition, the finding of Balantidium and Sarcocystis may have zoonotic implications and can therefore be used as markers of public health importance.

  20. First report of cerebellar abiotrophy in an Arabian foal from Argentina

    Science.gov (United States)

    Sadaba, S.A.; Madariaga, G.J.; Botto, C.M. Corbi; Carino, M.H.; Zappa, M.E.; García, P. Peral; Olguín, S.A.; Massone, A.; Díaz, S.

    2016-01-01

    Evidence of cerebellar abiotrophy (CA) was found in a six-month-old Arabian filly with signs of incoordination, head tremor, wobbling, loss of balance and falling over, consistent with a cerebellar lesion. Normal hematology profile blood test and cerebrospinal fluid analysis excluded infectious encephalitis, and serological testing for Sarcocystis neurona was negative. The filly was euthanized. Postmortem X-ray radiography of the cervical cephalic region identified not abnormalities, discounting spinal trauma. The histopathological analysis of serial transverse cerebellar sections by electron microscopy revealed morphological characteristics of apoptotic cells with pyknotic nuclei and degenerate mitochondria, cytoplasmic condensation and areas with absence of Purkinje cells, matching with CA histopathological characteristics. The indirect DNA test for CA was positive in the filly, and DNA test confirmed the CA carrier state in the parents and the recessive inheritance of the disease. To our knowledge this is the first report of a CA case in Argentina. PMID:28116251

  1. Prevalence of Zoonotic and Other Intestinal Protozoan Parasites in Stray Cats (Felis domesticus of Kerman, South-East of Iran

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    Sakineh BEIGI

    2017-01-01

    Full Text Available Intestinal protozoan parasites constitute a major source of diseases for stray cats and have been recognized as important public health problems in several parts of the world. Considering the potential risk of stray cats for public health, present cross-sectional study was carried out to determine the type and frequency of protozoan parasites by faecal examination. A total of 100 stray cats were examined in Kerman city, Iran, Overall 67 cats (67% were infected with at least one protozoan parasite. The following parasites, with their respective prevalence, were found; Isospora felis 38%, Isospora rivolta 25%, Toxoplasma gondii 16%, Sarcocystis spp. 8%, Cryptosporidium spp. 7%, and Giardia sp. 5%. Based on our data, the sex of stray cats was not significantly associated with the prevalence of gastrointestinal protozoan parasites. The high infection rate of zoonotic intestinal protozoan parasites in stray cats is considered to be critical from the viewpoint of public health importance.We

  2. First report of cerebellar abiotrophy in an Arabian foal from Argentina

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    S.A. Sadaba

    2016-12-01

    Full Text Available Evidence of cerebellar abiotrophy (CA was found in a six-month-old Arabian filly with signs of incoordination, head tremor, wobbling, loss of balance and falling over, consistent with a cerebellar lesion. Normal hematology profile blood test and cerebrospinal fluid analysis excluded infectious encephalitis, and serological testing for Sarcocystis neurona was negative. The filly was euthanized. Postmortem X-ray radiography of the cervical cephalic region identified not abnormalities, discounting spinal trauma. The histopathological analysis of serial transverse cerebellar sections by electron microscopy revealed morphological characteristics of apoptotic cells with pyknotic nuclei and degenerate mitochondria, cytoplasmic condensation and areas with absence of Purkinje cells, matching with CA histopathological characteristics. The indirect DNA test for CA was positive in the filly, and DNA test confirmed the CA carrier state in the parents and the recessive inheritance of the disease. To our knowledge this is the first report of a CA case in Argentina.

  3. Diagnóstico diferencial de trombose aortoilíaca e mieloencefalite protozoária equina: relato de caso Differential diagnosis between aorto-iliac thrombosis and equine protozoal myeloencephalitis: case report

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    P.B. Escodro

    2010-10-01

    Full Text Available Relata-se o caso de uma égua de atividade de polo, que apresentou inicialmente claudicação leve no membro posterior esquerdo, a qual evoluiu para ataxia e atrofia da musculatura glútea do lado esquerdo, com diagnóstico de trombose aortoilíaca (TAI. A paciente foi tratada com suspeita de mieloencefalite protozoária equina, devido à semelhança dos sinais clínicos com essa doença, porém o líquido cefalorraquidiano apresentou-se negativo para anticorpos anti-Sarcocystis neurona. A palpação transretal indicou uma massa na bifurcação aortoilíaca esquerda. Na avaliação ultrassonográfica, visualizou-se imagem hiperecoica aderida ao endotélio vascular, sugerindo TAI atingindo a estenose de 70% da luz arterial.The case of a mare used for polo is reported. The animal showed clinical signs of soft lameness of the hindlimb, evolving to ataxia and gluteal muscle atrophy, with aorto-iliac thrombosis (AIT. The patient was treated with the suspect of equine protozoal myeloencephalitis (EPM, due to the resemblance of clinical signs. Cerebrospinal fluid analysis was negative for antibodies against Sarcocystis neurona. The transrectal examination indicated a mass in the left aorto-iliac bifurcation. In the ultrasonographic evaluation, a hyperechoic image adhered to the vascular endothelium was observed, suggesting (AIT, occupying 70% of arterial lumen. The present article has the objective of pointing out the importance of the differential diagnosis between AIT and EPM in horses with ataxia in hindlimbs and muscular atrophy.

  4. Protozoan and helminth parasite fauna of free-living Croatian wild wolves (Canis lupus) analyzed by scat collection.

    Science.gov (United States)

    Hermosilla, Carlos; Kleinertz, Sonja; Silva, Liliana M R; Hirzmann, Jörg; Huber, Djuro; Kusak, Josip; Taubert, Anja

    2017-01-15

    The European wolf (Canis lupus) is a large carnivore species present in limited areas of Europe with several small populations still being considered as endangered. Wolves can be infected by a wide range of protozoan and metazoan parasites with some of them affecting free-living wolf health condition. On this account, an epidemiological survey was conducted to analyze the actual parasite fauna in Croatian wild wolves. In total, 400 individual faecal samples were collected during field studies on wolf ecology in the years 2002-2011. Parasite stages were identified by the sodium acetate acetic acid formalin (SAF)-technique, carbolfuchsin-stained faecal smears and Giardia/Cryptosporidium coproantigen-ELISAs. A subset of taeniid eggs-positive wolf samples was additionally analyzed by PCR and subsequent sequencing to identify eggs on Echinococcus granulosus/E. multilocularis species level. In total 18 taxa of parasites were here detected. Sarcocystis spp. (19.1%) occurred most frequently in faecal samples, being followed by Capillaria spp. (16%), ancylostomatids (13.1%), Crenosoma vulpis (4.6%), Angiostrongylus vasorum (3.1%), Toxocara canis (2.8%), Hammondia/Neospora spp. (2.6 %), Cystoisospora ohioensis (2.1%), Giardia spp. (2.1%), Cystoisospora canis (1.8%), Cryptosporidium spp. (1.8%), Trichuris vulpis (1.5%), Taenia spp. (1.5%), Diphyllobothrium latum (1.5%), Strongyloides spp. (0.5%), Opisthorchis felineus (0.5%), Toxascaris leonina (0.3%), Mesocestoides litteratus (0.3%) and Alaria alata (0.3%). Some of the here identified parasites represent relevant pathogens for wolves, circulating between these carnivorous definitive hosts and a variety of mammalian intermediate hosts, e. g. Taenia spp. and Sarcocystis spp., while others are considered exclusively pathogenic for canids (e.g. A. vasorum, C. vulpis, T. vulpis, Cystoisospora spp.). This study provides first records on the occurrence of the two relevant anthropozoonotic parasites, Giardia spp. and Cryptosporidium

  5. Survey of Dogs’ Parasites in Khorasan Razavi Province, Iran

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    GhR Razmi

    2009-12-01

    Full Text Available "nBackground: Dog is known to act as definitive host for some parasites that cause important diseases in man and animals. The aim of the present study was to determine the prevalence of Neospora caninum and other intestinal parasites in dogs in Khorasan Razavi Province, Iran. "nMethods: A cross-sectional study was done concerning frequency of N. canium and other in­testinal parasites in dogs in Mashhad area. Totally, 174 fecal samples from 89 farm dogs and 85 household dogs were collected from 2006 to 2007. Fecal samples were examined for de­tecting intestinal parasites by Mini Parasep®SF faecal parasite concentrator in Department of Parasitology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Iran "nResults: The overall prevalence of other intestinal parasites in farm dogs and household dogs were 29.21% and 14.11%, respectively. Seven parasites were found in farm dogs as follows: Toxocara canis 17.9%, Taenia sp. 10.1% , Strongyloides stercoralis 5.6%, Hammondia Neo­spora-like oocysts (HNLO 4.4% , Isospora sp. 7.8 %, Sarcocystis sp. 7.8 % and   Giardia sp. 1.1%  and four parasite in housed dogs:  Toxocara. 4.4%, Taenia sp. 3.3 % , Isospora sp. 2.3 % and  Sarcocystis sp. 4.7 %.  The fecal samples with HNLO were examined by N. caninum -specific PCR, and two of samples were positive for N. caninum. "nConclusion: The farm and household dogs are the source of some important zoonotic and non-zoonotic diseases in Iran .

  6. [Parasitological fecal studies of equids, dogs, cats and hedgehogs during the years 1984-1991].

    Science.gov (United States)

    Epe, C; Ising-Volmer, S; Stoye, M

    1993-11-01

    The results of the coproscopical examinations in horses, dogs, cats and hedgehogs between 1984 and 1991 are presented. In 9192 samples from horses 55.5% stages of strongylids, 4.0% of Parascaris equorum, 2.2% of anoplocephalids, 1.6% Strongyloides westeri, 0.7% of Oxyuris equi, 0.6% of Eimeria leuckarti, 0.2% of Fasciola hepatica and 0.04% of Dictyocaulus arnfieldi were found. In 48.0% of the 46 samples from donkeys eggs from strongylids were detected, in 17.4% larvae from Dictyocaulus arnfieldi, in 2.2% eggs from Strongyloides westeri, Parascaris equorum and oocysts from Eimeria leuckarti, respectively. In 3329 samples of dogs 6.9% developmental stages of Toxocara canis, 6.0% of Giardia spp., 4.2% of Isospora spp., 3.0% of Sarcocystis spp., 2.5% each of ancylostomids and Trichuris vulpis, 1.1% of Toxascaris leonina and 1.1% of Dipylidium canium, up to 1.0% of taeniids, 0.6% of each Mesocestoides spp. and Metastrongylidae, 0.3% of Strongyloides stercoralis and 0.2% of Capillaria spp. and Hammondia heydorni were detected. In 9.5% of the 1147 samples of cats eggs from Toxocara mystax were found, in 4.7% eggs of taeniids, in 4.6% cysts of Isospora spp., in 2.4% of Giardia spp., in 1.4% eggs of Dipylidium caninum, in 1.0% of Capillaria spp. and Aelurostrongylus abstrusus, in 0.6% development stages of Toxoplasma gondii, in 0.5% of ancylostomids and in 0.3% of Sarcocystis spp. and Opisthorchis felineus. In 1175 samples of hedgehogs 48.8% eggs of Capillaria spp., 35.9% of Crenosoma striatum, 17.9% oocysts of Isospora spp., 2.3% eggs of Brachylaemus erinacei were found.

  7. Microscopic and molecular characterization of Hepatozoon domerguei (Apicomplexa and Foleyella furcata (Nematoda in wild endemic reptiles from Madagascar

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    Maia João P.

    2014-01-01

    Full Text Available Madagascar is one of the world’s top twelve “megadiversity” hot spots hosting unique and threatened flora and fauna. Parasites are a major component of biodiversity but remain largely uncharacterized in wildlife. In this study we combine microscopic and molecular assessment of hemoparasites in endemic reptile species from Madagascar. We detected three distinct parasites: the apicomplexans Hepatozoon and Sarcocystis, and filarial nematodes. The prevalence and intensity of these apicomplexans were low overall, while microfilarial infections in chameleons were relatively high. We detected mixed infections of two Hepatozoon haplotypes in Madagascarophis colubrinus, and of Hepatozoon and microfilariae in a Furcifer sp. Phylogenetic analyses of Hepatozoon showed evidence of prey-predator transmission, with identical sequences found in the snakes M. colubrinus and Ithycyphus oursi, and their prey Furcifer sp. Based on previous studies regarding the life cycle of Hepatozoon domerguei Landau, Chabaud, Michel, and Brygoo, 1970 in these hosts and due to their morphological similarity, we propose that this Hepatozoon haplotype is Hepatozoon domerguei. Future studies, including the examination of invertebrate hosts, are needed to verify this preliminary taxonomic identification. A distinct hemogregarine haplotype was found in Oplurus sp., which displayed morphologically different gametocytes, some of which were apparently inside leukocytes. The Sarcocystis identified from Tracheloptychus petersi was identical to that reported in a North African snake, indicating that the same lineage is found in geographically distinct regions. By combining morphological and genetic information, Foleyella furcata (Linstow, 1899 filarial nematodes were identified in several Furcifer chameleons. This study provides insights into the distribution, diversity and host-parasite interactions of hemoparasites in wild reptile populations from Madagascar.

  8. Brief quarantine technology instructions of Sporozoosis%住肉孢子虫病检疫技术概述

    Institute of Scientific and Technical Information of China (English)

    张雅为; 魏澍; 闫海滨; 朱江巍

    2013-01-01

      住肉孢子虫病是由住肉孢子虫引起的一种原虫病,本病发生于各种家畜(马、牛、羊、猪、兔等)、鼠类、鸟类、爬虫类和鱼类中,偶尔也寄生于人。我国各地的牛羊,尤其是南方的水牛和黄牛、北方的绵羊和山羊常有发现,有些地区的感染率可达100%。住肉孢子虫寄生于各种家畜的横纹肌,致使肌肉不能食用,引起巨大的经济损失。本文对住肉孢子虫病的产地检疫技术、屠宰检疫技术和实验室检疫技术作以简要介绍。%Sporozoosis is a kind of protozoosis caused by sarcocystis sp. It can oc-cur in livestock such as horse, cattle, sheep, swine, rabbit, muroid, birds, reptil-ia and fish. Occasionally occurred in human. Cattle and sheep, especially buffalo and cattle in southern area in China, and sheep and goat in north area in China are more often positive of sporozoosis, in some area the posistive rate can be achieved 100%. Sarcocystis can be found in striated muscle of livestock, cause a huge econom-ic lost. In this article producing area quarantine technology, slaughting quarantine technology and lab quarantine technology of sporozoosis is described, providing use-ful technology.

  9. The prevalence of intestinal parasites in dogs from Prague, rural areas, and shelters of the Czech Republic.

    Science.gov (United States)

    Dubná, S; Langrová, I; Nápravník, J; Jankovská, I; Vadlejch, J; Pekár, S; Fechtner, J

    2007-04-10

    The prevalence of intestinal parasites was evaluated by examination of dog faecal samples in the Prague city centre, agricultural areas, and two shelters. The overall prevalence of parasites (i.e., protozoa and helminths, mentioned below) in Prague was 17.6%. Toxocara canis was the most common parasite, and was recovered from 6.2% of dogs, followed by Cystoisospora spp. (2.4%), Cryptosporidium spp. (1.4%), Trichuris sp. (1.1%), Taenia-type (1.0%), Giardia spp. (0.1%), Toxascaris sp. (0.9%), Dipylidium sp. (0.7%), Sarcocystis spp. (0.6%), Capillaria spp. (0.6%), Neospora/Hammondia spp. (0.5%), Ancylostoma sp. (0.4%), Uncinaria sp. (0.4%), and Spirocerca sp. (0.2%). The prevalence of infections with helminths and protozoans in two animal shelters in Prague was examined at the dog's admittance ir reception to the shelters and during housing. T. canis eggs (6.5%), Cystoisospora (4.4%), and Giardia (3.3%) cysts were the most prevalent. Significant increases in the prevalence of some parasites were found after a stay in the shelter. Giardia spp. showed an 11-fold increase in prevalence of dogs placed in the shelters for a longer time; Cryptosporidium spp. had a 7-fold increase, Capillaria spp. a 5-fold, Spirocerca sp., Neospora/Hammondia spp., and Cystoisospora spp. a 4-fold increase over dogs examined at the time of admittance to the shelter (pcanis was 13.7%, followed by Cystoisospora spp. (8.0%), Taenia spp. (3.5%), Sarcocystis spp. (3.0%), Giardia spp. (2.2%), Cryptosporidium spp. (2.0%), Trichuris sp. (1.7%), Toxascaris sp. (1.7%), Dipylidium sp. (1.3%), Neospora/Hammondia spp. (1.3%), Spirocerca sp. (1.1%), Uncinaria sp. (0.9%), Ancylostoma sp. (0.7%), and Capillaria spp. (0.6%). Examinations of dogs in urban and rural areas showed, with the exception of Trichuris sp. in Prague, a higher occurrence of nematode infection in autumn, notably T. canis (chi2>8.3, d.f.=3, p<0.04).

  10. Prevalence and Risk Factors of Intestinal Parasites in Cats from China

    Science.gov (United States)

    2015-01-01

    The prevalence of intestinal parasites in cats from China was largely unknown prior to this study. The aim of the present study was to investigate the presence of intestinal parasites in cats from central China and also identify risk factors for parasitism. Fecal samples from 360 cats were examined using sugar flotation procedure and fecal smear test by microscope. Cats had mixed two or three kinds of parasites infections. Of the 360 cats feces, intestinal parasites positive feces were 149 (41.39%). 64 (17.78%) were infected with Toxocara cati, 61 (16.94%) with Isospora felis, 41 (11.39%) with Isospora rivolta, 33 (9.17%) with Paragonimus, 23 (6.39%) with hookworms, 11 (3.06%) with Toxoplasma-like oocysts, 10 (2.78%) with Trichuris, 4 (1.11%) with lungworm, 2 (0.56%) with Sarcocystis, and 1 (0.28%) with Trematode. The cats' living outdoor was identified as risk factor by statistical analysis. These results provide relevant basic data for assessing the infection of intestinal parasites in cats from central region of China. In conclusion, there was high prevalence of intestinal parasites in cats from China. PMID:26078975

  11. Prevalence and Risk Factors of Intestinal Parasites in Cats from China

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    Yurong Yang

    2015-01-01

    Full Text Available The prevalence of intestinal parasites in cats from China was largely unknown prior to this study. The aim of the present study was to investigate the presence of intestinal parasites in cats from central China and also identify risk factors for parasitism. Fecal samples from 360 cats were examined using sugar flotation procedure and fecal smear test by microscope. Cats had mixed two or three kinds of parasites infections. Of the 360 cats feces, intestinal parasites positive feces were 149 (41.39%. 64 (17.78% were infected with Toxocara cati, 61 (16.94% with Isospora felis, 41 (11.39% with Isospora rivolta, 33 (9.17% with Paragonimus, 23 (6.39% with hookworms, 11 (3.06% with Toxoplasma-like oocysts, 10 (2.78% with Trichuris, 4 (1.11% with lungworm, 2 (0.56% with Sarcocystis, and 1 (0.28% with Trematode. The cats’ living outdoor was identified as risk factor by statistical analysis. These results provide relevant basic data for assessing the infection of intestinal parasites in cats from central region of China. In conclusion, there was high prevalence of intestinal parasites in cats from China.

  12. Pork as a source of human parasitic infection.

    Science.gov (United States)

    Djurković-Djaković, O; Bobić, B; Nikolić, A; Klun, I; Dupouy-Camet, J

    2013-07-01

    Foodborne zoonoses have been estimated to annually affect 10% of the global population, among which zoonotic parasites constitute an important class of aetiological agents. The major meatborne parasites include the protozoa Toxoplasma gondii and Sarcocystis spp., and the helminths Trichinella spp. and Taenia spp., all of which may be transmitted by pork. The significance of zoonotic parasites transmitted by pork consumption is emphasized by the prediction by the Food and Agriculture Organization of an 18.5% increase in world pork production over the next 10 years. Of all the porkborne parasites, the three 'T' parasites have been responsible for most porkborne illness throughout history; they are still endemic, and therefore are important public-health concerns, in developing countries. Although the risk of porkborne parasites, particularly helminths, may currently be considered insignificant in developed countries, the modern trend of consuming raw meat favours their re-emergence. This paper overviews the main parasites transmitted to humans by pork, and outlines the main lines of prevention.

  13. Endoparasites in dogs and cats in Germany 1999-2002.

    Science.gov (United States)

    Barutzki, D; Schaper, R

    2003-07-01

    Infections with endoparasites in dogs and cats have been determined by analysing the results of faecal examinations (Flotation, MIFC, sedimentation, Baermann, smear, ProSpecT Giardia Microplate Assay). Samples of 8438 dogs and 3167 cats from the years 1999 until 2002 have been included in the investigation. 2717 dogs (32.2%) and 771 cats (24.3%) have been infected with endoparasites. In the infected dogs the following parasites have been identified: Class Nematodea: Toxocara canis: 22.4%, Toxascaris leonina: 1.8%, Ancylostomatidae: 8.6%, Trichuris vulpis: 4.0%, Capillaria spp.: 2.3%, Crenosoma vulpis: 0.9%, Angiostrongylus vasorum: 0.3%; Class Cestodea: Taeniidae: 1.2%, Dipylidium caninum: 0.4%, Diplopylidium/Joyeuxiella: 0.1%, Mesocestoides: 0.2%, Diphyllobothrium latum: canis: 8.0%, C. ohioensis: 17.0%, Hammondia/Neospora: 1.7%; Class Zoomastigophorea: Giardia spp.: 51.6%. In the 771 infected cats the following prevalences of parasites have been found: Class Nematodea: Toxocara mystax: 26.2%, Ancylostoma tubaeforme: 0.3%, Capillaria spp.: 7.0%, Aelurostrongylus abstrusus: 2.7%; Class Cestodea: Taeniidae: 2.6%, Dipylidium caninum: 0.1%; Class Sporozoea: Sarcocystis spp.: 2.2%, Cystoisospora spp.: 21.9%, C. felis: 15.3%, C. rivolta: 7.9%, Toxoplasma/Hammondia: 4.5%; Class Zoomastigophorea: Giardia spp.: 51.6%.

  14. Specific detection of Neospora caninum oocysts in fecal samples from experimentally-infected dogs using the polymerase chain reaction.

    Science.gov (United States)

    Hill, D E; Liddell, S; Jenkins, M C; Dubey, J P

    2001-04-01

    Neospora caninum oocysts, passed in the feces of a definitive host (dog), were isolated, and genomic DNA was extracted. A polymerase cahin reaction (PCR) targeting the N. caninum-specific Nc 5 genomic sequence was performed using the isolated DNA. A synthesized competitor molecule containing part of the Nc 5 sequence was included in the assay as a check against false-negative PCR results and to quantify N. caninum oocyst DNA in fecal samples. A standard curve of the ratio of fluorescence intensity of PCR-amplified competitor to that of oocyst DNA was constructed to compare oocyst equivalents from fecal samples containing unknown numbers of N. caninum oocysts and to assess the sensitivity of the assay. The specificity of the assay was determined using the Nc 5-specific primers in PCR assays against other parasites likely to be found in canine feces. Genomic DNA sequences from the canine coccidians Hammondia heydorni, Cryptosporidium parvum, Sarcocystis cruzi, S. tenella, and Isospora ohioensis and the canine helminth parasites Strongyloides stercoralis, Toxocara canis, Dipylidium caninum, and Ancylostoma caninum were not amplified. In addition, genomic DNA sequences from oocysts of coccidian parasites that might contaminate dog feces, such as Hammondia hammondi, Toxoplasma gondii, or Eimeria tenella, were not amplified in the PCR assay. The assay should be useful in epidemiological surveys of both domestic and wild canine hosts and in investigations of oocyst biology in experimental infections.

  15. Estimation of canine intestinal parasites in Córdoba (Spain) and their risk to public health.

    Science.gov (United States)

    Martínez-Moreno, F J; Hernández, S; López-Cobos, E; Becerra, C; Acosta, I; Martínez-Moreno, A

    2007-01-19

    The prevalence of gastrointestinal parasites in dogs was studied in the province of Córdoba (southern Spain), with special attention to those parasites that can be transmitted to man. The experiment was completed with the examination of soil samples from public parks and city gardens. The study was carried out over a population of 1800 animals entered in the Control Animal Centre (CECA) by coprological methods, and within this group, 300 dogs were sacrificed and necropsied. The prevalence of any intestinal parasitic infection was 71.33%. The following parasites of the gastrointestinal tract were recorded: Isospora canis (22%), Isospora (Cystoisospora) spp. (10.22%), Sarcocystis (2.5%), Hammondia/Neospora (1.94%), Giardia canis (1%), Dipylidium caninum (13.2%), Taenia hydatigena (7.66%), Taenia pisiformis (4%), Uncinaria stenocephala (33.27%), Toxascaris leonina (14.94%), Toxocara canis (17.72%) and Trichuris vulpis (1.66%). Related to public health, it is important to point out the presence of T. canis only in puppies younger than one year and Uncinaria, more frequent in adult dogs. Soil samples of parks revealed the presence of eggs of Toxocara, and it suggests the existence of real risk for human infection.

  16. RNG1 is a Late Marker of the Apical Polar Ring in Toxoplasma gondii

    Science.gov (United States)

    Tran, Johnson Q.; de Leon, Jessica C.; Li, Catherine; Huynh, My-Hang; Beatty, Wandy; Morrissette, Naomi S.

    2010-01-01

    The asexually proliferating stages of apicomplexan parasites cause acute symptoms of diseases such as malaria, cryptosporidiosis and toxoplasmosis. These stages are characterized by the presence of two independent microtubule organizing centers (MTOCs). Centrioles are found at the poles of the intranuclear spindle. The apical polar ring (APR), a MTOC unique to apicomplexans, organizes subpellicular microtubules which impose cell shape and apical polarity on these protozoa. Here we describe the characteristics of a novel protein that localizes to the APR of Toxoplasma gondii which we have named ring-1 (RNG1). There are related RNG1 proteins in Neospora caninum and Sarcocystis neurona but no obvious homologs in Plasmodium spp., Cryptosporidium spp. or Babesia spp. RNG1 is a small, low-complexity, detergent-insoluble protein that assembles at the APR very late in the process of daughter parasite replication. We were unable to knock-out the RNG1 gene, suggesting that its gene product is essential. Tagged RNG1 lines have also allowed us to visualize the APR during growth of Toxoplasma in the microtubule-disrupting drug oryzalin. Oryzalin inhibits nuclear division and cytokinesis although Toxoplasma growth continues, and similar to earlier observations of unchecked centriole duplication in oryzalin-treated parasites, the APR continues to duplicate during aberrant parasite growth. PMID:20658557

  17. Prey choice and habitat use drive sea otter pathogen exposure in a resource-limited coastal system

    Science.gov (United States)

    Johnson, Christine K.; Tinker, M. Tim; Estes, James A.; Conrad, Patricia A.; Staedler, Michelle M.; Miller, Melissa A.; Jessup, David A.; Mazet, Jonna A.K.

    2014-01-01

    The processes promoting disease in wild animal populations are highly complex, yet identifying these processes is critically important for conservation when disease is limiting a population. By combining field studies with epidemiologic tools, we evaluated the relationship between key factors impeding southern sea otter (Enhydra lutris nereis) population growth: disease and resource limitation. This threatened population has struggled to recover despite protection, so we followed radio-tagged sea otters and evaluated infection with 2 disease-causing protozoal pathogens, Toxoplasma gondii and Sarcocystis neurona, to reveal risks that increased the likelihood of pathogen exposure. We identified patterns of pathogen infection that are linked to individual animal behavior, prey choice, and habitat use. We detected a high-risk spatial cluster of S. neurona infections in otters with home ranges in southern Monterey Bay and a coastal segment near San Simeon and Cambria where otters had high levels of infection with T. gondii. We found that otters feeding on abalone, which is the preferred prey in a resource-abundant marine ecosystem, had a very low risk of infection with either pathogen, whereas otters consuming small marine snails were more likely to be infected with T. gondii. Individual dietary specialization in sea otters is an adaptive mechanism for coping with limited food resources along central coastal California. High levels of infection with protozoal pathogens may be an adverse consequence of dietary specialization in this threatened species, with both depleted resources and disease working synergistically to limit recovery.

  18. DISCOVERY OF THREE NOVEL COCCIDIAN PARASITES INFECTING CALIFORNIA SEA LIONS (ZALOPHUS CALIFORNIANUS), WITH EVIDENCE OF SEXUAL REPLICATION AND INTERSPECIES PATHOGENICITY

    Science.gov (United States)

    Colegrove, Kathleen M.; Grigg, Michael E.; Carlson-Bremer, Daphne; Miller, Robin H.; Gulland, Frances M. D.; Ferguson, David J. P.; Rejmanek, Daniel; Barr, Bradd C.; Nordhausen, Robert; Melli, Ann C.; Conrad, Patricia A.

    2016-01-01

    Enteric protozoal infection was identified in 5 stranded California sea lions (Zalophus californianus). Microscopically, the apical cytoplasm of distal jejunal enterocytes contained multiple stages of coccidian parasites, including schizonts with merozoites and spherical gametocytes, which were morphologically similar to coccidians. By histopathology, organisms appeared to be confined to the intestine and accompanied by only mild enteritis. Using electron microscopy, both sexual (microgametocytes, macrogamonts) and asexual (schizonts, merozoites) coccidian stages were identified in enterocytes within parasitophorous vacuoles, consistent with apicomplexan development in a definitive host. Serology was negative for tissue cyst-forming coccidians, and immunohistochemistry for Toxoplasma gondii was inconclusive and negative for Neospora caninum and Sarcocystis neurona. Analysis of ITS-1 gene sequences amplified from frozen or formalin-fixed paraffin-embedded intestinal sections identified DNA sequences with closest homology to Neospora sp. (80%); these novel sequences were referred to as belonging to coccidian parasites ‘‘A,’’ ‘‘B,’’ and ‘‘C.’’ Subsequent molecular analyses completed on a neonatal harbor seal (Phoca vitulina) with protozoal lymphadenitis, hepatitis, myocarditis, and encephalitis showed that it was infected with a coccidian parasite bearing the ‘‘C’’ sequence type. Our results indicate that sea lions likely serve as definitive hosts for 3 newly described coccidian parasites, at least 1 of which is pathogenic in a marine mammal intermediate host species. PMID:21495828

  19. Gene Discovery in the Apicomplexa as Revealed by EST Sequencing and Assembly of a Comparative Gene Database

    Science.gov (United States)

    Li, Li; Brunk, Brian P.; Kissinger, Jessica C.; Pape, Deana; Tang, Keliang; Cole, Robert H.; Martin, John; Wylie, Todd; Dante, Mike; Fogarty, Steven J.; Howe, Daniel K.; Liberator, Paul; Diaz, Carmen; Anderson, Jennifer; White, Michael; Jerome, Maria E.; Johnson, Emily A.; Radke, Jay A.; Stoeckert, Christian J.; Waterston, Robert H.; Clifton, Sandra W.; Roos, David S.; Sibley, L. David

    2003-01-01

    Large-scale EST sequencing projects for several important parasites within the phylum Apicomplexa were undertaken for the purpose of gene discovery. Included were several parasites of medical importance (Plasmodium falciparum, Toxoplasma gondii) and others of veterinary importance (Eimeria tenella, Sarcocystis neurona, and Neospora caninum). A total of 55,192 ESTs, deposited into dbEST/GenBank, were included in the analyses. The resulting sequences have been clustered into nonredundant gene assemblies and deposited into a relational database that supports a variety of sequence and text searches. This database has been used to compare the gene assemblies using BLAST similarity comparisons to the public protein databases to identify putative genes. Of these new entries, ∼15%–20% represent putative homologs with a conservative cutoff of p neurona: , , , , , , , , , , , , , –, –, –, –, –. Eimeria tenella: –, –, –, –, –, –, –, –, – , –, –, –, –, –, –, –, –, –, –, –. Neospora caninum: –, –, , – , –, –.] PMID:12618375

  20. Enteric bacterial pathogen detection in southern sea otters (Enhydra lutris nereis) is associated with coastal urbanization and freshwater runoff

    Science.gov (United States)

    Miller, Melissa A.; Byrne, Barbara A.; Jang, Spencer S.; Dodd, Erin M.; Dorfmeier, Elene; Harris, Michael D.; Ames, Jack; Paradies, David; Worcester, Karen; Jessup, David A.; Miller, Woutrina A.

    2009-01-01

    Although protected for nearly a century, California’s sea otters have been slow to recover, in part due to exposure to fecally-associated protozoal pathogens like Toxoplasma gondii and Sarcocystis neurona. However, potential impacts from exposure to fecal bacteria have not been systematically explored. Using selective media, we examined feces from live and dead sea otters from California for specific enteric bacterial pathogens (Campylobacter, Salmonella, Clostridium perfringens, C. difficile and Escherichia coli O157:H7), and pathogens endemic to the marine environment (Vibrio cholerae, V. parahaemolyticus and Plesiomonas shigelloides). We evaluated statistical associations between detection of these pathogens in otter feces and demographic or environmental risk factors for otter exposure, and found that dead otters were more likely to test positive for C. perfringens, Campylobacter and V. parahaemolyticus than were live otters. Otters from more urbanized coastlines and areas with high freshwater runoff (near outflows of rivers or streams) were more likely to test positive for one or more of these bacterial pathogens. Other risk factors for bacterial detection in otters included male gender and fecal samples collected during the rainy season when surface runoff is maximal. Similar risk factors were reported in prior studies of pathogen exposure for California otters and their invertebrate prey, suggesting that land-sea transfer and/or facilitation of pathogen survival in degraded coastal marine habitat may be impacting sea otter recovery. Because otters and humans share many of the same foods, our findings may also have implications for human health. PMID:19720009

  1. Prevalence of Protozoa and Gastrointestinal Helminthes in Stray Cats in Zanjan Province, North-West of Iran

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    SA Altome

    2009-07-01

    Full Text Available Background: Cats and other felines act as definitive hosts for many intestinal parasites, some of which are responsible for several zoonotic diseases.  The aim of this study was to determine the type and prevalence of protozoa and gastrointestinal helminthes among stray cats. Methods: A cross sectional study was conducted. Digestive tracts of 100 stray cats in Zanjan Province, north-west of Iran were autopsied in order to recognize gastrointestinal helminthes and intestinal protozoan parasites. These cats were collected by baited cage trapped from October 2007 to September 2008. Gender and species of helminthes and protozoa were rec­ognized using authentic diagnostic criteria. Statistical evaluation was performed by SPSS version 14. Results: Forty-two percent of cats were infected with intestinal protozoan parasites, 33% were infected with cestodes and 39% infected with gastrointestinal nematodes. Four species protozoan parasites and eight gastrointestinal helminthes were recovered from the animals, including Taenia taeniaeformis, Dipylidium spp., Joyeuxiella pasqaulei, Toxocara cati, Phy­saloptera praeputialis, Rectalaria spp., Onicolla, Cystoisospora spp., Toxoplasma gondii, and Sarcocystis spp . Conclusions: The high infection rate of Toxoplasma and some gastrointestinal helminthes in stray cats is considered to be critical from the viewpoint of public health importance.

  2. Prevalence and Risk Factors of Intestinal Parasites in Cats from China.

    Science.gov (United States)

    Yang, Yurong; Liang, Hongde

    2015-01-01

    The prevalence of intestinal parasites in cats from China was largely unknown prior to this study. The aim of the present study was to investigate the presence of intestinal parasites in cats from central China and also identify risk factors for parasitism. Fecal samples from 360 cats were examined using sugar flotation procedure and fecal smear test by microscope. Cats had mixed two or three kinds of parasites infections. Of the 360 cats feces, intestinal parasites positive feces were 149 (41.39%). 64 (17.78%) were infected with Toxocara cati, 61 (16.94%) with Isospora felis, 41 (11.39%) with Isospora rivolta, 33 (9.17%) with Paragonimus, 23 (6.39%) with hookworms, 11 (3.06%) with Toxoplasma-like oocysts, 10 (2.78%) with Trichuris, 4 (1.11%) with lungworm, 2 (0.56%) with Sarcocystis, and 1 (0.28%) with Trematode. The cats' living outdoor was identified as risk factor by statistical analysis. These results provide relevant basic data for assessing the infection of intestinal parasites in cats from central region of China. In conclusion, there was high prevalence of intestinal parasites in cats from China.

  3. Seasonal and biogeographical patterns of gastrointestinal parasites in large carnivores: wolves in a coastal archipelago.

    Science.gov (United States)

    Bryan, Heather M; Darimont, Chris T; Hill, Janet E; Paquet, Paul C; Thompson, R C Andrew; Wagner, Brent; Smits, Judit E G

    2012-05-01

    Parasites are increasingly recognized for their profound influences on individual, population and ecosystem health. We provide the first report of gastrointestinal parasites in gray wolves from the central and north coasts of British Columbia, Canada. Across 60 000 km(2), wolf feces were collected from 34 packs in 2005-2008. At a smaller spatial scale (3300 km(2)), 8 packs were sampled in spring and autumn. Parasite eggs, larvae, and cysts were identified using standard flotation techniques and morphology. A subset of samples was analysed by PCR and sequencing to identify tapeworm eggs (n=9) and Giardia cysts (n=14). We detected ≥14 parasite taxa in 1558 fecal samples. Sarcocystis sporocysts occurred most frequently in feces (43·7%), followed by taeniid eggs (23·9%), Diphyllobothrium eggs (9·1%), Giardia cysts (6·8%), Toxocara canis eggs (2·1%), and Cryptosporidium oocysts (1·7%). Other parasites occurred in ≤1% of feces. Genetic analyses revealed Echinococcus canadensis strains G8 and G10, Taenia ovis krabbei, Diphyllobothrium nehonkaiense, and Giardia duodenalis assemblages A and B. Parasite prevalence differed between seasons and island/mainland sites. Patterns in parasite prevalence reflect seasonal and spatial resource use by wolves and wolf-salmon associations. These data provide a unique, extensive and solid baseline for monitoring parasite community structure in relation to environmental change.

  4. Theileria infection in domestic ruminants in northern Ethiopia.

    Science.gov (United States)

    Gebrekidan, Hagos; Hailu, Asrat; Kassahun, Aysheshm; Rohoušová, Iva; Maia, Carla; Talmi-Frank, Dalit; Warburg, Alon; Baneth, Gad

    2014-02-24

    Piroplasmosis caused by different tick-borne hemoprotozoan parasites of the genera Theileria and Babesia is among the most economically important infections of domestic ruminants in sub-Saharan Africa. A survey for piroplasm infection was conducted in three locations in Northern Ethiopia. Of 525 domestic ruminants surveyed, 80% of the cattle, 94% of the sheep and 2% of the goats were positive for different Theileria spp. based on PCR of blood followed by DNA sequencing. Sheep had a significantly higher rate of infection compared with cattle (PTheileria were detected in cattle: T. velifera, T. mutans, T. orientalis complex and T. annulata with infection rates of 66, 8, 4, and 2%, respectively. This is the first report of T. annulata, the cause of Tropical Theileriosis in Ethiopia. Of the two Theileria spp. detected in small ruminants, T. ovis was highly prevalent (92%) in sheep and rare in goats (1.5%) whereas T. seperata was infrequent in sheep (2%) and rare in goats (0.4%). None of the animals were positive for Babesia spp.; however, Sarcocystis capracanis and S. tenella were detected in one goat and a sheep, respectively. The widespread distribution of Theileria spp. among cattle in northern Ethiopia including the virulent T. annulata and more mildly pathogenic T. mutans and T. orientalis, and the high infection rate in sheep with the usually sub-clinical T. ovis indicate extensive exposure to ticks and transmission of piroplasms with an important economic impact.

  5. Enteric protozoa of cats and their zoonotic potential-a field study from Austria.

    Science.gov (United States)

    Hinney, Barbara; Ederer, Christina; Stengl, Carina; Wilding, Katrin; Štrkolcová, Gabriela; Harl, Josef; Flechl, Eva; Fuehrer, Hans-Peter; Joachim, Anja

    2015-05-01

    Domestic cats can be infected with a variety of enteric protozoa. Genotyping of protozoan species, especially Giardia as the most common, can improve assessment of their relevance as zoonotic agents. For an overview on the occurrence of feline enteric protozoa, 298 faecal samples of cats from private households, catteries and animal shelters in Austria were collected. All samples were examined by flotation and using a rapid test for Giardia (FASTest). For the detection of Tritrichomonas blagburni, freshly voided faeces (n = 40) were processed using a commercial culturing system (InPouch TF-Feline). Genotyping was done at the β-giardin gene loci (each sample) and triosephosphate isomerase gene loci (positive samples) for Giardia and at the 18S rRNA gene (positive samples) for Cryptosporidium. Thirty-seven samples (12.4%) were positive for Giardia by flotation and/or using a rapid test. Cryptosporidium was present in 1.7%, Cystoisospora in 4.0%, Sarcocystis in 0.3% and T. blagburni in 2.5% of the samples. Genotyping revealed Giardia cati, the potentially zoonotic Giardia duodenalis and Cryptosporidium felis. Most of the infected cats had no diarrhoea. Cats from shelters were significantly more often infected than owned cats (p = 0.01). When comparing Giardia detection methods, the rapid test had a higher sensitivity than flotation. Polymerase chain reaction (PCR) results were mostly independent from the other two tests.

  6. Sarcosporidiose muscular: registro de um caso

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    João Aris Kouyoumdjian

    1985-09-01

    Full Text Available Um caso de sarcosporidiose muscular em uma paciente de 29 anos de idade é relatado. O quadro clínico evidenciava síndrome miopática tipo cinturas com leve mialgia de 4 anos de evolução. A biópsia muscular do músculo deltóide mostrou a presença de formações císticas de permeio a fibras musculares normais. O diâmetro, comprimento e provável cápsula dessas formações são características da infestação pelo coccídeo Sarcocystis sp. Outras características histopatológi-cas e a distinção com as formações císticas da toxoplasmose são comentadas. A maioria dos casos descritos na literatura - pouco mais de três dezenas - evidencia a ausência de quadro clínico típico, devendo a moléstia entrar no diagnóstico diferencial ocasional das doenças musculares esqueléticas ou cardíacas, inflamatórias e/ou parasitárias.

  7. Review of zoonotic parasites in medical and veterinary fields in the Republic of Korea.

    Science.gov (United States)

    Youn, Heejeong

    2009-10-01

    Zoonotic parasites are animal parasites that can infect humans. The major zoonotic protozoa in the Republic of Korea are Babesia bovis, Chilomastix mesnili, Cryptosporidium parvum, Endolimax nana, Entamoeba coli, Entamoeba hitolytica, Giardia lamblia, Iodamoeba bütschlii, Pneumocystis carinii, Sarcocystis cruzi, and Toxoplasma gondii. The major zoonotic helminths in Korea include trematodes, cestodes, and nematodes. Trematodes are Clonorchis sinensis, Echinostoma hortense, Echinostoma spp., Fasciola hepatica, Heterophyes nocens, Metagonimus yokogawai, and Paragonimus westermani. Cestodes are Diphyllobothrium latum, Dipylidium caninum, Echinococcus granulosus, Hymenolepis nana, Raillietina tetragona, sparganum (Spirometra spp.), Taenia saginata, T. solium, and T. asiatica. Nematodes are Ancylostoma caninum, Brugia malayi, Capillaria hepatica, Dirofilaria immitis, Gnathostoma dololesi, Gnathostoma spinigerum, Loa loa, Onchocerca gibsoni, Strongyloides stercoralis, Thelazia callipaeda, Trichinella spiralis, Trichostrongylus orientalis, Trichuris trichiura, and Trichuris vulpis. The one arthropod is Sarcoptes scabiei. Many of these parasites have disappeared or were in decline after the 1990's. Since the late 1990's, the important zoonotic protozoa have been C. parvum, E. nana, E. coli, E. hitolytica, G. lamblia, I. buetschlii, P. carinii and T. gondii. The important zoonotic helminths have been C. sinensis, H. nocens, M. yokogawai, P. westermani, D. latum, T. asiatica, sparganum, B. malayi, T. orientalis, T. callipaeda and T. spiralis. However, outbreaks of these parasites are only in a few endemic areas. The outbreaks of Enterobius vermicularis and head lice, human parasites, have recently increased in the kindergartens and primary schools in the Republic of Korea.

  8. Intestinal parasites of the red fox (Vulpes vulpes) in Slovenia.

    Science.gov (United States)

    Vergles Rataj, Aleksandra; Posedi, Janez; Zele, Diana; Vengušt, Gorazd

    2013-12-01

    In the present study, 428 foxes were collected and examined for intestinal helminths using the washing-out method. Parasites were found in 93.2% of the examined animals. The most frequently identified nematodes were Uncinaria stenocephala (58.9%), Toxocara canis (38.3%) and Molineus patens (30.6%). Other nematodes found were Pterygodermatites affinis (4.2%), Capillaria sp. (2.8%), Crenosoma vulpis (2.8%), Toxascaris leonina (2.5%), Trichuris vulpis (0.7%) and Physaloptera sp. (0.2%). Mesocestoides sp. (27.6%) and Taenia crassiceps (22.2%) were the most prevalent cestodes, followed by T. polyacantha (6.5%), Hymenolepis nana (2.1%), T. pisiformis (2.1%) and Dipylidium caninum (1.4%). The study also revealed four trematode species: Rossicotrema donicum (1.6%), Heterophyes heterophyes (1.1%), Metagonimus yokogawai (1.1%), Prohemistomum appendiculatum (0.4%) and two protozoan species: oocysts of Sarcocystis (2.8%) and Isospora (0.4%). This is the first extensive study on the intestinal parasites of the red fox (Vulpes vulpes) in Slovenia. The 2.6% prevalence of Echinococcus multilocularis in the same sample population as investigated herein has been reported previously (Vergles Rataj et al., 2010).

  9. Parasitic Contamination of Raw Vegetables in Zanjan Markets, Iran

    Directory of Open Access Journals (Sweden)

    Negin Torabi

    2016-09-01

    Full Text Available Background: Complex surface of vegetables facilitate attachment and transmission of several pathogens. No previous study has been conducted in survey of parasitic contamination of vegetables in Zanjan. This study aimed to detect the parasitic contamination in common raw vegetables in Zanjan markets. Methods: A total of 352 raw vegetable samples, including leek, parsley, basil, mint, radish, cress and dill were collected from grocery stores using cluster sampling in different regions of the city during 2014. The edible parts of vegetables were separated and immersed in normal saline solution. Floating vegetables were removed and the solution was allowed to sediment at room temperature for 24 hours. The pellet was examined following sedimentation and floatation methods. Results:Various Organisms were detected in 54% (190 of the 352 samples, but only 2.8% of samples had pathogenic parasites including; Trichostrongylus eggs (3, Hookworm eggs (2, Eimeria oocysts (2, Sarcocystis oocyst (1, Strongyloides larvae (1, and Fasciola eggs (1. The contamination rate of vegetables was highest (90.4% in the fall (p˂0.05. Conclusion: Vegetable contamination with parasitic organisms in this area was low, maybe due to irrigation of vegetables with sources other than sewage water, but it is still necessary to improve sanitary conditions of vegetables.

  10. Population expansion and individual age affect endoparasite richness and diversity in a recolonising large carnivore population

    Science.gov (United States)

    Lesniak, Ines; Heckmann, Ilja; Heitlinger, Emanuel; Szentiks, Claudia A.; Nowak, Carsten; Harms, Verena; Jarausch, Anne; Reinhardt, Ilka; Kluth, Gesa; Hofer, Heribert; Krone, Oliver

    2017-01-01

    The recent recolonisation of the Central European lowland (CEL) by the grey wolf (Canis lupus) provides an excellent opportunity to study the effect of founder events on endoparasite diversity. Which role do prey and predator populations play in the re-establishment of endoparasite life cycles? Which intrinsic and extrinsic factors control individual endoparasite diversity in an expanding host population? In 53 individually known CEL wolves sampled in Germany, we revealed a community of four cestode, eight nematode, one trematode and 12 potential Sarcocystis species through molecular genetic techniques. Infections with zoonotic Echinococcus multilocularis, Trichinella britovi and T. spiralis occurred as single cases. Per capita endoparasite species richness and diversity significantly increased with population size and changed with age, whereas sex, microsatellite heterozygosity, and geographic origin had no effect. Tapeworm abundance (Taenia spp.) was significantly higher in immigrants than natives. Metacestode prevalence was slightly higher in ungulates from wolf territories than from control areas elsewhere. Even though alternative canid definitive hosts might also play a role within the investigated parasite life cycles, our findings indicate that (1) immigrated wolves increase parasite diversity in German packs, and (2) prevalence of wolf-associated parasites had declined during wolf absence and has now risen during recolonisation. PMID:28128348

  11. Population expansion and individual age affect endoparasite richness and diversity in a recolonising large carnivore population

    Science.gov (United States)

    Lesniak, Ines; Heckmann, Ilja; Heitlinger, Emanuel; Szentiks, Claudia A.; Nowak, Carsten; Harms, Verena; Jarausch, Anne; Reinhardt, Ilka; Kluth, Gesa; Hofer, Heribert; Krone, Oliver

    2017-01-01

    The recent recolonisation of the Central European lowland (CEL) by the grey wolf (Canis lupus) provides an excellent opportunity to study the effect of founder events on endoparasite diversity. Which role do prey and predator populations play in the re-establishment of endoparasite life cycles? Which intrinsic and extrinsic factors control individual endoparasite diversity in an expanding host population? In 53 individually known CEL wolves sampled in Germany, we revealed a community of four cestode, eight nematode, one trematode and 12 potential Sarcocystis species through molecular genetic techniques. Infections with zoonotic Echinococcus multilocularis, Trichinella britovi and T. spiralis occurred as single cases. Per capita endoparasite species richness and diversity significantly increased with population size and changed with age, whereas sex, microsatellite heterozygosity, and geographic origin had no effect. Tapeworm abundance (Taenia spp.) was significantly higher in immigrants than natives. Metacestode prevalence was slightly higher in ungulates from wolf territories than from control areas elsewhere. Even though alternative canid definitive hosts might also play a role within the investigated parasite life cycles, our findings indicate that (1) immigrated wolves increase parasite diversity in German packs, and (2) prevalence of wolf-associated parasites had declined during wolf absence and has now risen during recolonisation.

  12. Enteric protozoan parasites in stray cats in Kuwait with special references to toxoplasmosis and risk factors affecting its occurrence.

    Science.gov (United States)

    Abdou, Nadra-Elwgoud M I; Al-Batel, Maha K; El-Azazy, Osama M E; Sami, Attia M; Majeed, Qais A H

    2013-08-01

    In Kuwait, stray cats were surveyed for enteric protozoan infection using fecal examination and their sera were tested for Toxoplasma gondii IgG using indirect hemagglutination test (IHAT) as well as for feline immunodeficiency virus (FIV) antibodies and feline leukaemia virus (FeLV) antibodies using ELISA. Out of 240 fecal samples examined 22 (9.2%) were found to be infected with oocysts of four species of coccidian protozoa. Isopspora felis was the most predominant enteric protozoan parasite (7.1%), followed by T. gondii (2.1%), I. rivolta (1.6), Sarcocystis was only found in one case (0.4%). Juvenile cats ( 6 months old) had higher infection rate with oocyst of enteric protozoa than older cats (p-value 0.001). Sero-survey of 240 stray cats revealed that 19.6% were positive to T. gondii IgG. Toxoplasma sero-positivity was observed in higher number of adults compared to young cats suggests that with age the risk of exposure to T. gondii increases. While concurrent retroviral infections were not found to be associated with increased risk for developing T. gondii antibodies.

  13. Experimental Infection of Swine by Isospora suis Biester 1934 for Species Confirmation

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    Silvia Maria Oliveira Sayd

    1998-11-01

    Full Text Available A survey of Isospora suis performed in 177 faecal samples from 30 swine farms detected thin wall type I. suis oocysts in seven samples. This type of oocyst measuring 23.9 by 20.7 mm had a retracted thin wall similar to that of the genus Sarcocystis. This type of oocysts, isolated from four different faecal samples, was inoculated in four-five-days-old piglets free of contamination in order to verify the life cycle and pathogenicity of the species. The pigs were kept in individual metal cages and fed with cow milk. Daily faecal collections and examinations were performed until the 21st day after infection. MacMaster and Sheather' s methods were used for oocyst counting and identification. Infected piglets produced yellowish-pasty diarrhoea with slight dehydration. The prepatent and patent periods were respectively from 6 to 9 and 3 to 10 days after infection. Oocyst elimination was interrupted on the 10th and 11th days after infection with biphasic cycles. Thin and thick wall oocysts were detected in the same faecal samples. Thin walls were not observed in unsporulated oocysts. The observations suggest that this type of oocysts could appear in specific strains which occur in the later stages of their development. These oocysts seem to be responsible for clinical and pathogenic signs of neonatal isosporosis in pigs.

  14. Parasitic zoonoses in Papua New Guinea.

    Science.gov (United States)

    Owen, I L

    2005-03-01

    Relatively few species of zoonotic parasites have been recorded in humans in Papua New Guinea. A greater number of potentially zoonotic species, mostly nematodes, occur in animals but are yet to be reported from humans. Protozoa is the best represented group of those infecting man, with Giardia duodenalis, Cryptosporidium parvum, Cyclospora cayetanesis, Toxoplasma gondii, Sarcocystis spp., Entamoeba polecki, Balantidium coli and, possibly, Blastocystis hominis. The only zoonotic helminths infecting humans include the trematode Paragonimus westermani, the cestodes Hymenolepis nana, H. diminuta and the sparganum larva of Spirometra erinacea, and the nematodes Trichinella papuae and Angiostrongylus cantonensis and, possibly, Ascaris suum. Other groups represented are Acanthocephala (Macracanthorhynchus hirudinaceus)), insects (Chrysomya bezziana, Cimex sp., Ctenocephalides spp.), and mites (Leptotrombidium spp. and, possibly Sarcoptes scabiei, and Demodex sp.). One leech (Phytobdella lineata) may also be considered as being zoonotic. The paucity of zoonotic parasite species can be attributed to long historical isolation of the island of New Guinea and its people, and the absence until recent times of large placental mammals other than pig and dog. Some zoonotic helminths have entered the country with recent importation of domestic animals, in spite of quarantine regulations, and a few more (two cestodes, one nematode and one tick) are poised to enter from neighbouring countries, given the opportunity. Improvement in water supplies, human hygiene and sanitation would reduce the prevalence of many of these parasites, and thorough cooking of meat would lessen the risk of infection by some others.

  15. Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses Diagnóstico e isolamento de Toxoplasma gondii em equídeos de frigoríficos brasileiros

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    Fernanda Evers

    Full Text Available This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64, and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6% by IFAT and in 14 (3.5% by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16, T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15% of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed to T. gondii.O objetivo do estudo foi investigar anticorpos anti-Toxoplasma gondii e isolar o parasita do cérebro de equídeos abatidos em matadouros-frigoríficos no Brasil. Colheram-se amostras de 398 cérebros e sangue de equídeos machos e fêmeas de idades variadas, provenientes de seis estados brasileiros. As amostras de soro foram avaliadas pelo teste de imunofluorescência indireta (IFI para T. gondii (ponto de corte ≥ 64, e os fragmentos de cérebros foram submetidos ao bioensaio em camundongos. Por meio da IFI, 46 (11,6% equídeos foram soropositivos. Pelo bioensaio em camundongos, 14 (3,5% cérebros de equídeos testados foram positivos. Em doze dos bioensaios, os camundongos foram positivos somente pela IFI (ponto

  16. Cytokine gene signatures in neural tissue of horses with equine protozoal myeloencephalitis or equine herpes type 1 myeloencephalopathy.

    Science.gov (United States)

    Pusterla, N; Wilson, W D; Conrad, P A; Barr, B C; Ferraro, G L; Daft, B M; Leutenegger, C M

    2006-09-09

    This study was designed to determine the relative levels of gene transcription of selected pathogens and cytokines in the brain and spinal cord of 12 horses with equine protozoal myeloencephalitis (EPM), 11 with equine herpesvirus type 1 (EHV-1) myeloencephalopathy, and 12 healthy control horses by applying a real time pcr to the formalin-fixed and paraffin-embedded tissues. Total rna was extracted from each tissue, transcribed to complementary dna (cDNA) and assayed for Sarcocystis neurona, Neospora hughesi, EHV-1, equine GAPDH (housekeeping gene), tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10 AND IL-12 p40. S neurona cdna was detected in the neural tissue from all 12 horses with EPM, and two of them also had amplifiable cDNA of N hughesi. The relative levels of transcription of protozoal cdna ranged from 1 to 461 times baseline (mean 123). All the horses with ehv-1 myeloencephalopathy had positive viral signals by PCR with relative levels of transcription ranging from 1 to 1618 times baseline (mean 275). All the control horses tested negative for S neurona, N hughesi and EHV-1 cdna. The cytokine profiles of each disease indicated a balance between pro- and anti-inflammatory markers. In the horses with epm the pro-inflammatory Th1 cytokines (IL-8, TNF-alpha and IFN-gamma) were commonly expressed but the anti-inflammatory Th2 cytokines (IL-4, IL-6 AND IL-10) were absent or rare. In the horses with ehv-1 the proinflammatory cytokine IL-8 was commonly expressed, but IL-10 and IFN-gamma were not, and TNF-alpha was rare. Tissue from the control horses expressed only the gene GAPDH.

  17. Polyparasitism is associated with increased disease severity in Toxoplasma gondii-infected marine sentinel species.

    Directory of Open Access Journals (Sweden)

    Amanda K Gibson

    2011-05-01

    Full Text Available In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91% of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42% and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies

  18. Detection and characterization of diverse coccidian protozoa shed by California sea lions

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    Yvette A. Girard

    2016-04-01

    Full Text Available Tissue-cyst forming coccidia in the family Sarcocystidae are etiologic agents of protozoal encephalitis in marine mammals including the federally listed Southern sea otter (Enhydra lutris. California sea lions (Zalophus californianus, whose coastal habitat overlaps with sea otters, are definitive hosts for coccidian protozoa provisionally named Coccidia A, B and C. While Coccidia A and B have unknown clinical effects on aquatic wildlife hosts, Coccidia C is associated with severe protozoal disease in harbor seals (Phoca vitulina. In this study, we conducted surveillance for protozoal infection and fecal shedding in hospitalized and free-ranging California sea lions on the Pacific Coast and examined oocyst morphology and phenotypic characteristics of isolates via mouse bioassay and cell culture. Coccidia A and B were shed in similar frequency, particularly by yearlings. Oocysts shed by one free-ranging sea lion sampled at Año Nuevo State Park in California were previously unidentified in sea lions and were most similar to coccidia infecting Guadalupe fur seals (Arctocephalus townsendi diagnosed with protozoal disease in Oregon (USA. Sporulated Coccidia A and B oocysts did not replicate in three strains of mice or in African green monkey kidney cells. However, cultivation experiments revealed that the inoculum of fecally-derived Coccidia A and B oocysts additionally contained organisms with genetic and antigenic similarity to Sarcocystis neurona; despite the absence of detectable free sporocysts in fecal samples by microscopic examination. In addition to the further characterization of Coccidia A and B in free-ranging and hospitalized sea lions, these results provide evidence of a new role for sea lions as putative mechanical vectors of S. neurona, or S. neurona-like species. Future work is needed to clarify the distribution, taxonomical status, and pathogenesis of these parasites in sea lions and other marine mammals that share their the near

  19. Results of parasitological examinations of faecal samples from cats and dogs in Germany between 2003 and 2010.

    Science.gov (United States)

    Barutzki, Dieter; Schaper, Roland

    2011-08-01

    In a retrospective study, the results of parasitological examinations of faecal samples from 8,560 cats and 24,677 dogs between January 2003 and December 2010 in Germany were analysed. 30.4 % of the examined dogs and 22.8 % of the cats were infected with endoparasites. The examination of the faecal samples from dogs revealed stages of Giardia spp. (18.6 %), Toxocara canis (6.1 %), Toxascaris leonina (0.6 %), Ancylostomatidae (2.2 %), Trichuris vulpis (1.2 %), Capillaria spp. (1.3 %), Crenosoma vulpis (0.4 %), Angiostrongylus vasorum (0.5 %), Taeniidae (0.4 %), Dipylidiidae (canis (2.4 %), Sarcocystis spp. (2.2 %) and Hammondia heydorni/Neospora caninum (0.3 %). Dogs in the age groups up to 3 months and > 3 up to 6 months of age showed significantly higher infection rates with Giardia spp. (37.5 % and 38.2 %, respectively), Toxocara canis (12.0 % and 12.4 %, respectively), Toxascaris leonina (1.1 % and 1.6 %, respectively), Isospora spp. (23.4 % and 11.8 %, respectively), I. ohioensis-complex (15.6 % and 7.2 %, respectively) and I. canis (11.8 % and 5.2 %, respectively) compared to older dogs. In faecal samples from cats, stages of Giardia spp. (12.6 %), Toxocara cati (4.7 %), Toxascaris leonina (0.1 %), Ancylostoma tubaeforme (0.2 %), Aelurostrongylus abstrusus (0.5 %), Capillaria spp. (1.0 %), Taeniidae (0.6 %), Dipylidium caninum ( 3 up to 6 months of age showed significantly higher infection rates with Giardia spp. (19.5 % and 24.0 %, respectively), T. cati (8.1 % and 6.9 %, respectively), Isospora spp. (12.8 % and 8.6 %, respectively), I. felis (10.0 % and 5.9%, respectively) and I. rivolta (4.6 % and 2.9%, respectively) compared to older cats.

  20. SURVEY FOR INFECTIOUS DISEASE IN THE SOUTH AMERICAN FUR SEAL (ARCTOCEPHALUS AUSTRALIS) POPULATION AT PUNTA SAN JUAN, PERU.

    Science.gov (United States)

    Jankowski, Gwen; Adkesson, Michael J; Saliki, Jeremiah T; Cárdenas-Alayza, Susana; Majluf, Patricia

    2015-06-01

    The Peruvian population of the South American fur seal ( Arctocephalus australis ) is a distinct evolutionarily significant unit that is endangered. One of the largest rookeries for this species in Peru is located within the Punta San Juan marine protected area (15°22'S, 75°12'W). To better understand the current health status of this population, exposure to 10 pinniped pathogens was evaluated in adult female fur seals (n=29) via serology and polymerase chain reaction (PCR) techniques in November 2010. The results suggest this population is naïve to canine and phocine distemper viruses (serum neutralization test), five Leptospira interrogans serovars (microscopic agglutination test), and Brucella canis (card test). Indirect fluorescent antibody testing for Toxoplasma gondii , Neospora caninum , and Sarcocystis neurona was also uniformly negative. PCR testing of nasal swabs using previously described Mycoplasma spp. primers was positive in 37.9% (11/29) of samples. One animal was positive via card test for Brucella abortus , whereas 53.7% (15/28) were positive or suspect using a marine Brucella competitive enzyme-linked immunosorbent assay. Antibody to phocine herpesvirus-1 (PHV-1) was identified in 85.7% (24/28) of the sampled population by serum neutralization testing. Overall, exposure to Mycoplasma spp., Brucella spp., and PHV-1 was observed, but results demonstrated low to no exposure to many key pinniped pathogens. The expansion of human populations, agriculture, and industry along the Peruvian coast may lead to increased pathogen exposure from human, domestic, and wild animal sources. The naïve nature of this key population of South American fur seals raises concerns about potential risk for disease outbreaks.

  1. Assessment of clinical pathology and pathogen exposure in sea otters (Enhydra lutris) bordering the threatened population in Alaska

    Science.gov (United States)

    Goldstein, T.; Gill, V.A.; Tuomi, Pamela A.; Monson, Daniel H.; Burdin, A.; Conrad, P.A.; Dunn, J.L.; Field, C.; Johnson, Chad; Jessup, David A.; Bodkin, J.; Doroff, A.M.

    2011-01-01

    Northern sea otter (Enhydra lutris kenyoni) abundance has decreased dramatically over portions of southwest Alaska, USA, since the mid-1980s, and this stock is currently listed as threatened under the Endangered Species Act. In contrast, adjacent populations in south central Alaska, USA, and Russia have been stable to increasing during the same period. Sea otters bordering the area classified in the recent decline were live-captured during 2004–2006 at Bering Island, Russia, and the Kodiak Archipelago, Alaska, USA, to evaluate differences in general health and current exposure status to marine and terrestrial pathogens. Although body condition was lower in animals captured at Bering Island, Russia, than it was at Kodiak, USA, clinical pathology values did not reveal differences in general health between the two regions. Low prevalences of antibodies (>5%) were found in Kodiak, USA, and on Bering Island, Russia, to Toxoplasma gondii, Sarcocystis neurona, and Leptospira interrogans. Exposure to phocine herpesvirus-1 was found in both Kodiak, USA (15.2%), and Bering Island, Russia (2.3%). Antibodies to Brucella spp. were found in 28% of the otters tested on Bering Island, Russia, compared with only 2.7% of the samples from Kodiak, USA. Prevalence of exposure to Phocine distemper virus (PDV) was 41% in Kodiak, USA, but 0% on Bering Island, Russia. Archived sera from southwest and south-central Alaska dating back to 1989 were negative for PDV, indicating exposure occurred in sea otters in Kodiak, USA, in recent years. Because PDV can be highly pathogenic in naïve and susceptible marine mammal populations, tissues should be examined to explore the contribution of this virus to otter deaths. Our results reveal an increase in exposure to pathogens in sea otters in Kodiak, Alaska, USA, since the 1990s.

  2. Cytokine Gene Expression in Response to SnSAG1 in Horses with Equine Protozoal Myeloencephalitis

    Science.gov (United States)

    Spencer, Jennifer A.; Deinnocentes, Patricia; Moyana, Edith M.; Guarino, Anthony J.; Ellison, Siobhan E.; Bird, R. Curtis; Blagburn, Byron L.

    2005-01-01

    Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome seen in horses from the Americas and is mainly caused by Sarcocystis neurona. Recently, a 29-kDa surface antigen from S. neurona merozoites was identified as being highly immunodominant on a Western blot. This antigen has been sequenced and cloned, and the expressed protein has been named SnSAG1. In a previous study, cell-mediated immune responses to SnSAG1 were shown to be statistically significantly reduced in horses with EPM in comparison to EPM-negative control horses. It therefore appears as though the parasite is able to induce immunosuppression towards parasite-derived antigens as parasite-specific responses are decreased. Isolated peripheral blood lymphocytes from 21 EPM (cerebrospinal fluid [CSF] Western blot)-negative horses with no clinical signs and 21 horses with clinical signs of EPM (CSF Western blot positive) were cocultured with SnSAG1 for 48 and 72 h, and the effect on cytokine production was investigated by means of reverse transcriptase PCR. Cytokines assayed include gamma interferon (IFN-γ), tumor necrosis factor alpha, interleukin (IL)-2, IL-4, and IL-6. β-Actin was used as the housekeeping gene. A Wilcoxon signed-rank test of the findings indicated that there was a statistically significant decrease in IFN-γ production after 48 h in culture for samples from horses with clinical disease. There was also a statistically significant increase in IL-4 production after 72 h in culture for samples from horses with EPM. These results further support the notion that this parasite is able to subvert the immune system in horses with clinical disease. PMID:15879026

  3. Polyparasitism Is Associated with Increased Disease Severity in Toxoplasma gondii-Infected Marine Sentinel Species

    Science.gov (United States)

    Gibson, Amanda K.; Raverty, Stephen; Lambourn, Dyanna M.; Huggins, Jessica; Magargal, Spencer L.; Grigg, Michael E.

    2011-01-01

    In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species) as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals) sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91%) of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42%) and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies polyparasitism as

  4. Reduced Levels of Nitric Oxide Metabolites in Cerebrospinal Fluid Are Associated with Equine Protozoal Myeloencephalitis

    Science.gov (United States)

    Njoku, Chinedu J.; Saville, William J. A.; Reed, Stephen M.; Oglesbee, Michael J.; Rajala-Schultz, Päivi J.; Stich, Roger W.

    2002-01-01

    Equine protozoal myeloencephalitis (EPM) is a disease of horses that is primarily associated with infection with the apicomplexan Sarcocystis neurona. Infection with this parasite alone is not sufficient to induce the disease, and the mechanism of neuropathogenesis associated with EPM has not been reported. Nitric oxide (NO) functions as a neurotransmitter, a vasodilator, and an immune effector and is produced in response to several parasitic protozoa. The purpose of this work was to determine if the concentration of NO metabolites (NOx−) in the cerebrospinal fluid (CSF) is correlated with the development of EPM. CSF NOx− levels were measured before and after transport-stressed, acclimated, or dexamethasone-treated horses (n = 3 per group) were experimentally infected with S. neurona sporocysts. CSF NOx− levels were also compared between horses that were diagnosed with EPM after natural infection with S. neurona and horses that did not have clinical signs of disease or that showed no evidence of infection with the parasite (n = 105). Among the experimentally infected animals, the mean CSF NOx− levels of the transport-stressed group, which had the most severe clinical signs, was reduced after infection, while these values were found to increase after infection in the remaining groups that had less severe signs of EPM. Under natural conditions, horses with EPM (n = 65) had a lower mean CSF NOx− concentration than clinically normal horses with antibodies (Abs) against S. neurona (n = 15) in CSF, and horses that developed ataxia (n = 81) had a significantly lower mean CSF NOx− concentration than horses that did not have neurologic signs (n = 24). In conclusion, lower CSF NOx− levels were associated with clinical EPM, suggesting that measurement of CSF NOx− levels could improve the accuracy of diagnostic tests that are based upon detection of S. neurona-specific Abs in CSF alone and that reduced NO levels could be causatively related to the development

  5. Detection and characterization of diverse coccidian protozoa shed by California sea lions

    Science.gov (United States)

    Girard, Yvette A.; Johnson, Christine K.; Fritz, Heather M.; Shapiro, Karen; Packham, Andrea E.; Melli, Ann C.; Carlson-Bremer, Daphne; Gulland, Frances M.; Rejmanek, Daniel; Conrad, Patricia A.

    2015-01-01

    Tissue-cyst forming coccidia in the family Sarcocystidae are etiologic agents of protozoal encephalitis in marine mammals including the federally listed Southern sea otter (Enhydra lutris). California sea lions (Zalophus californianus), whose coastal habitat overlaps with sea otters, are definitive hosts for coccidian protozoa provisionally named Coccidia A, B and C. While Coccidia A and B have unknown clinical effects on aquatic wildlife hosts, Coccidia C is associated with severe protozoal disease in harbor seals (Phoca vitulina). In this study, we conducted surveillance for protozoal infection and fecal shedding in hospitalized and free-ranging California sea lions on the Pacific Coast and examined oocyst morphology and phenotypic characteristics of isolates via mouse bioassay and cell culture. Coccidia A and B were shed in similar frequency, particularly by yearlings. Oocysts shed by one free-ranging sea lion sampled at Año Nuevo State Park in California were previously unidentified in sea lions and were most similar to coccidia infecting Guadalupe fur seals (Arctocephalus townsendi) diagnosed with protozoal disease in Oregon (USA). Sporulated Coccidia A and B oocysts did not replicate in three strains of mice or in African green monkey kidney cells. However, cultivation experiments revealed that the inoculum of fecally-derived Coccidia A and B oocysts additionally contained organisms with genetic and antigenic similarity to Sarcocystis neurona; despite the absence of detectable free sporocysts in fecal samples by microscopic examination. In addition to the further characterization of Coccidia A and B in free-ranging and hospitalized sea lions, these results provide evidence of a new role for sea lions as putative mechanical vectors of S. neurona, or S. neurona-like species. Future work is needed to clarify the distribution, taxonomical status, and pathogenesis of these parasites in sea lions and other marine mammals that share their the near-shore marine

  6. Prevalence of zoonotic intestinal parasites in domestic and stray dogs in a rural area of Iran.

    Science.gov (United States)

    Beiromvand, Molouk; Akhlaghi, Lame; Fattahi Massom, Seyed Hossein; Meamar, Ahmad Reza; Motevalian, Abbas; Oormazdi, Hormozd; Razmjou, Elham

    2013-04-01

    Certain zoonotic parasites are enteropathogens in dogs that cause serious human disease such as cystic echinococcosis, human alveolar echinococcosis, visceral larva migrans, and ocular larva migrans. This study investigated the prevalence of intestinal parasites in dogs in the Chenaran County, Razavi Khorasan Province, Iran. Sampling was carried out randomly in 17 villages from November 2009 to January 2010. Seventy-seven fecal samples from 28 domestic and 49 stray dogs were examined using sieving/flotation and modified Ziehl-Neelsen staining. Intestinal parasites were found in 51 of the 77 (66%) dogs most common being Toxascaris leonina (29%, 22/77), Toxocara spp. (25%, 19/77), Eimeria spp. (19%, 15/77), Taenia/Echinococcus spp. (18%, 14/77), Sarcocystis spp. (17%, 13/77), and Dicrocoelium dendriticum (14%, 11/77). Lower infection rates of parasites were observed for Trichuris vulpis (6%, 5/77), Cryptosporidium spp. (5%, 4/77), and Physaloptera spp. (3%, 2/77). Prevalence of infection by Dipylidium caninum, Capillaria spp., Cystoisospora spp., and hookworms was similar (1%, 1/77). This study is the first report of the prevalence of intestinal parasites of domestic and stray dogs in Chenaran County, Northeast Iran. The higher prevalence of zoonotic intestinal parasites such as Toxascaris leonina, Toxocara spp. and Taenia/Echinococcus spp. compared to other parasites indicates the need for control programs to minimize the risk of transmission of zoonotic disease, particularly cystic echinococcosis, alveolar echinococcosis, visceral larva migrans, and ocular larva migrans to people living in these areas.

  7. Protein tyrosine nitration in chronic intramuscular parasitism: immunohistochemical evaluation of relationships between nitration, and fiber type-specific responses to infection

    Directory of Open Access Journals (Sweden)

    Ted H. Elsasser

    2011-05-01

    Full Text Available The present study was conducted to determine whether preferential muscle catabolism [psoas major (PM > rectus femoris (RF] observed during the chronic intramuscular stage of Sarcocystis cruzi infection could be associated with the pathological consequences of increased protein tyrosine nitration in fibers characteristically more metabolically active due to higher mitochondrial density. Holstein calves were assigned to control (C, or S. cruzi-infected (I groups, n=5/group. Calves were euthanized on day 63 of infection. Samples of RF and PM were prepared for metabolic fiber typing (MFT: slow oxidative, SO – Type I; fast oxidative glycolytic, FOG - Type IIa; fast glycolytic, FG – Type IIb, fiber area, and immunohistochemical localization of fast myosin heavy chain 2a and 2b, nitrotyrosine (NT, and mitochondrial Complex V ATP-synthase. MFT analysis documented that PM contained twice the number of SO fibers compared to RF (32 v 16%, P<0.002. SO and FOG fibers (Both higher in mitochondrial density than FG fibers in both PM and RF were significantly smaller in area in I calves with mean FG areas not different between C and I. Muscle NT content (Western blot of myofibrillar protein fraction increased with infection; NT was immunohistochemically localized into three distinct patterns in fibers: i sparse fiber staining, ii dense punctuate intrafiber staining, and iii pericystic staining. By image analysis, the greatest punctuate intrafiber pixel density of NT was associated with SO fibers from I calves with the NT colocalizing with mitochondrial Complex V – F1F0 ATP synthase. More fibers were positive for the colocalization in PM than RF (P<0.04. The data are consistent with the concept that fibers rich in mitochondria possessing more inherent oxidative energy capacity generate more nitrated proteins than glycolytic fibers and as such are more affected by the proinflammatory response to infections like Sarcocystosis.

  8. Pathogen exposure and blood chemistry in the Washington population of northern sea otters (Enhydra lutris kenyoni)

    Science.gov (United States)

    White, C. LeAnn; Schuler, Krysten L.; Thomas, Nancy J.; Webb, Julie L.; Saliki, Jeremiah T.; Ip, Hon S.; Dubey, J.P.; Frame, Elizabeth R.

    2013-01-01

    Northern sea otters (Enhydra lutris kenyoni) from Washington State, United States were evaluated in 2011 to determine health status and pathogen exposure. Antibodies to Brucella spp. (10%) and influenza A virus (23%) were detected for the first time in this population in 2011. Changes in clinical pathology values (serum chemistries), exposure to pathogens, and overall health of the population over the last decade were assessed by comparing 2011 data to the data collected on this population in 2001–2002. Several serum chemistry parameters were different between study years and sexes but were not clinically significant. The odds of canine distemper virus exposure were higher for otters sampled in 2001–2002 (80%) compared to 2011 (10%); likelihood of exposure significantly increased with age. Prevalence of exposure to Sarcocystis neurona was also higher in 2001–2002 (29%) than in 2011 (0%), but because testing methods varied between study years the results were not directly comparable. Exposure to Leptospira spp. was only observed in 2001–2002. Odds of Toxoplasma gondii exposure were higher for otters sampled in 2011 (97%) than otters in 2001–2002 (58%). Substantial levels of domoic acid (n = 2) and saxitoxin (n = 2) were found in urine or fecal samples from animals sampled in 2011. No evidence of calicivirus or Coxiella burnetii exposure in the Washington population of northern sea otters was found in either 2001–2002 or 2011. Changes in exposure status from 2001–2002 to 2011 suggest that the Washington sea otter population may be dealing with new disease threats (e.g., influenza) while also increasing their susceptibility to diseases that may be highly pathogenic in naïve individuals (e.g., canine distemper).

  9. Detection and characterization of diverse coccidian protozoa shed by California sea lions.

    Science.gov (United States)

    Girard, Yvette A; Johnson, Christine K; Fritz, Heather M; Shapiro, Karen; Packham, Andrea E; Melli, Ann C; Carlson-Bremer, Daphne; Gulland, Frances M; Rejmanek, Daniel; Conrad, Patricia A

    2016-04-01

    Tissue-cyst forming coccidia in the family Sarcocystidae are etiologic agents of protozoal encephalitis in marine mammals including the federally listed Southern sea otter (Enhydra lutris). California sea lions (Zalophus californianus), whose coastal habitat overlaps with sea otters, are definitive hosts for coccidian protozoa provisionally named Coccidia A, B and C. While Coccidia A and B have unknown clinical effects on aquatic wildlife hosts, Coccidia C is associated with severe protozoal disease in harbor seals (Phoca vitulina). In this study, we conducted surveillance for protozoal infection and fecal shedding in hospitalized and free-ranging California sea lions on the Pacific Coast and examined oocyst morphology and phenotypic characteristics of isolates via mouse bioassay and cell culture. Coccidia A and B were shed in similar frequency, particularly by yearlings. Oocysts shed by one free-ranging sea lion sampled at Año Nuevo State Park in California were previously unidentified in sea lions and were most similar to coccidia infecting Guadalupe fur seals (Arctocephalus townsendi) diagnosed with protozoal disease in Oregon (USA). Sporulated Coccidia A and B oocysts did not replicate in three strains of mice or in African green monkey kidney cells. However, cultivation experiments revealed that the inoculum of fecally-derived Coccidia A and B oocysts additionally contained organisms with genetic and antigenic similarity to Sarcocystis neurona; despite the absence of detectable free sporocysts in fecal samples by microscopic examination. In addition to the further characterization of Coccidia A and B in free-ranging and hospitalized sea lions, these results provide evidence of a new role for sea lions as putative mechanical vectors of S. neurona, or S. neurona-like species. Future work is needed to clarify the distribution, taxonomical status, and pathogenesis of these parasites in sea lions and other marine mammals that share their the near-shore marine

  10. Polyparasitism is associated with increased disease severity in Toxoplasma gondii-infected marine sentinel species.

    Science.gov (United States)

    Gibson, Amanda K; Raverty, Stephen; Lambourn, Dyanna M; Huggins, Jessica; Magargal, Spencer L; Grigg, Michael E

    2011-05-01

    In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species) as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals) sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91%) of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42%) and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies polyparasitism as

  11. Sarcocystosis of chital-dhole: conditions for evolutionary stability of a predator parasite mutualism

    Directory of Open Access Journals (Sweden)

    Watve Milind G

    2005-02-01

    Full Text Available Abstract Background For parasites with a predator-prey life cycle, the completion of the life cycle often depends on consumption of parasitized prey by the predator. In the case of such parasite species the predator and the parasite have common interests and therefore a mutualistic relationship is possible. Some evidence of a predator-parasite mutualism was reported from spotted deer or chital (Axix axis as a prey species, dhole or Indian wild-dog (Cuon alpinus as the predator and a protozoan (Sarcocystis axicuonis as the parasite. We examine here, with the help of a model, the ecological conditions necessary for the evolution and stability of such a mutualistic relationship. A two – level game theory model was designed in which the payoff of a parasite is decided not only by alternative parasite strategies but also by alternative host strategies and vice versa. Conditions for ESS were examined. Results A tolerant predator strategy and a low or moderately virulent parasite strategy which together constitute mutualism are stable only at a high frequency of recycling of parasite and a substantial prey – capture benefit to the predator. Unlike the preliminary expectation, parasite will not evolve towards reduced virulence, but reach an optimum moderate level of virulence. Conclusion The available data on the behavioral ecology of dhole and chital suggest that they are likely to meet the stability criteria and therefore a predator-parasite mutualism can be stable in this system. The model also points out the gaps in the current data and could help directing further empirical work.

  12. A cross-sectional study on intestinal parasitic infections in rural communities, northeast Thailand.

    Science.gov (United States)

    Boonjaraspinyo, Sirintip; Boonmars, Thidarut; Kaewsamut, Butsara; Ekobol, Nuttapon; Laummaunwai, Porntip; Aukkanimart, Ratchadawan; Wonkchalee, Nadchanan; Juasook, Amornrat; Sriraj, Pranee

    2013-12-01

    Despite the existence of effective anthelmintics, parasitic infections remain a major public health problem in Southeast Asia, including Thailand. In rural communities, continuing infection is often reinforced by dietary habits that have a strong cultural basis and by poor personal hygiene and sanitation. This study presents a survey of the prevalence of intestinal parasitic infections among the people in rural Thailand. The community-based cross-sectional study was conducted in villages in Khon Kaen Province, northeastern Thailand, from March to August 2013. A total of 253 stool samples from 102 males and 140 females, aged 2-80 years, were prepared using formalin-ethyl acetate concentration methods and examined using light microscopy. Ninety-four individuals (37.2%) were infected with 1 or more parasite species. Presence of parasitic infection was significantly correlated with gender (P=0.001); nearly half of males in this survey (49.0%) were infected. Older people had a higher prevalence than younger members of the population. The most common parasite found was Opisthorchis viverrini (26.9%), followed by Strongyloides stercoralis (9.5%), Taenia spp. (1.6%), echinostomes (0.4%), and hookworms (0.4%). The prevalence of intestinal protozoa was Blastocystis hominis 1.6%, Entamoeba histolytica 0.8%, Entamoeba coli 0.8%, Balantidium coli 0.4%, Iodamoeba bütschlii 0.4%, and Sarcocystis hominis 0.4%. Co-infections of various helminths and protozoa were present in 15.9% of the people. The present results show that the prevalence of parasitic infections in this region is still high. Proactive education about dietary habits, personal hygiene, and sanitation should be provided to the people in this community to reduce the prevalence of intestinal parasite infections. Moreover, development of policies and programs to control parasites is needed.

  13. Protozoal Meningoencephalitis in Sea Otters (Enhydra lutris): a Histopathological and Immunohistochemical Study of Naturally Occuring Cases

    Science.gov (United States)

    Thomas, N.J.; Dubey, J.P.; Lindsay, D.S.; Cole, R.A.; Meteyer, C.U.

    2007-01-01

    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.

  14. First molecular characterization of enteric protozoa and the human pathogenic microsporidian, Enterocytozoon bieneusi, in captive snakes in China.

    Science.gov (United States)

    Karim, Md Robiul; Yu, Fuchang; Li, Jian; Li, Junqiang; Zhang, Longxian; Wang, Rongjun; Rume, Farzana Islam; Jian, Fuchun; Zhang, Sumei; Ning, Changshen

    2014-08-01

    Enteric protozoa are frequently found in snakes. Nevertheless, few studies regarding genetic characterization of these parasites have been carried out. We describe here the first molecular survey of protozoan pathogens from snakes in China and the first report on Enterocytozoon bieneusi genotyping in snakes in the world. Here, 240 fecal specimens were collected from two species of captive snakes, Naja naja (Indian cobra) and Ptyas mucosus (Oriental rat snake), in Guangxi Province, China, and examined by PCR amplification of the small subunit-ribosomal RNA of enteric protozoa and the internal transcribed spacer of ribosomal RNA of E. bieneusi. Cryptosporidium serpentis was identified in three specimens (2.1%) of Oriental rat snakes. Caryospora was found in 5.4% specimens, including eight from cobras (8.1%) and five from rat snakes (3.6%), and represented six new species-Caryospora sp. SKC-2014a to Caryospora sp. SKC-2014 f. Three new Eimeria species, Eimeria sp. SKE-2014a to Eimeria sp. SKE-2014c, were detected in three specimens (2.1%) from rat snakes. Additionally, Sarcocystis sp. SKS-2014 was detected in one specimen from a cobra. The infection rates of E. bieneusi were 3.0% in cobras and 5.7% in rat snakes. Sequence analysis of 11 PCR products revealed the presence of six E. bieneusi genotypes-two known genotypes (type IV and Henan V) and four new genotypes (CRep-1 to CRep-4). All six E. bieneusi genotypes belonged to the zoonotic group (group 1). This result raised the possibility that E. bieneusi could be present in animals consumed by snakes. This should be taken into consideration to better understand the diversity of the parasite, its transmission through the predator-prey relationship, and public health implications.

  15. Chagas disease in a Texan horse with neurologic deficits.

    Science.gov (United States)

    Bryan, Laura K; Hamer, Sarah A; Shaw, Sarah; Curtis-Robles, Rachel; Auckland, Lisa D; Hodo, Carolyn L; Chaffin, Keith; Rech, Raquel R

    2016-01-30

    A 10-year-old Quarter Horse gelding presented to the Texas A&M University Veterinary Teaching Hospital with a six month-history of ataxia and lameness in the hind limbs. The horse was treated presumptively for equine protozoal myeloencephalitis (EPM) based on clinical signs but was ultimately euthanized after its condition worsened. Gross lesions were limited to a small area of reddening in the gray matter of the thoracic spinal cord. Histologically, trypanosome amastigotes morphologically similar to Trypanosoma cruzi, the agent of Chagas disease in humans and dogs, were sporadically detected within segments of the thoracic spinal cord surrounded by mild lymphoplasmacytic inflammation. Ancillary testing for Sarcocystis neurona, Neospora spp., Toxoplasma gondii and Leishmania spp. was negative. Conventional and real time polymerase chain reaction (PCR) of affected paraffin embedded spinal cord were positive for T. cruzi, and sequencing of the amplified T. cruzi satellite DNA PCR fragment from the horse was homologous with various clones of T. cruzi in GenBank. While canine Chagas disease cases have been widely reported in southern Texas, this is the first report of clinical T. cruzi infection in an equid with demonstrable amastigotes in the spinal cord. In contrast to previous instances of Chagas disease in the central nervous system (CNS) of dogs and humans, no inflammation or T. cruzi amastigotes were detected in the heart of the horse. Based on clinical signs, there is a potential for misdiagnosis of Chagas disease with other infectious diseases that affect the equine CNS. T. cruzi should be considered as a differential diagnosis in horses with neurologic clinical signs and histologic evidence of meningomyelitis that originate in areas where Chagas disease is present. The prevalence of T. cruzi in horses and the role of equids in the parasite life cycle require further study. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. [Muscular sarcocystosis after travel to Malaysia: a case series from Germany].

    Science.gov (United States)

    Slesak, G; Tappe, D; Keller, C; Cramer, J; Güthoff, W; Zanger, P; Frank, M; Ernestus, K; Rauthe, S; Stich, A; Schäfer, J

    2014-05-01

    Since 2011, about 100 travellers to the island of Tioman, Malaysia, have been diagnosed worldwide with suspected muscular sarcocystosis, a previously only sporadically observed parasitic disease. Source of infection and therapy remain unclear. Final diagnosis requires microscopic identification of cysts in muscle biopsies. The study objective was a systematic description of characteristic symptoms, laboratory investigations and treatment response. Systematic case series. The 26 cases of 5 centers for tropical medicine in Germany showed a characteristic biphasic development: symptoms began in general 2 weeks after mid-holidays (min. 7.5, max. 22 days) with unspecific fever and headaches lasting for almost 1 week. After an asymptomatic period of 2 weeks, severe myalgia (6.5, scale 0-10) and fever developed and lasted for about 6 weeks (min. 7, max. 207 days), accompanied by creatin-phosphokinase(CK)-elevation (up to 3.5 times), and eosinophilia (2.9 times). One out of two muscle biopsies revealed a cyst typical for sarcocystosis. In 6 out of 7 patients an increase in Sarcocystis-specific antibody concentration could be demonstrated by ELISA. Treatment with systemic steroids and albendazole, or ivermectin resulted in significant symptomatic improvement in most of the patients. One patient was treated early with cotrimoxazole and subsequently did not develop a second phase of the disease. All patients had stayed in the North-West of the island Tioman. Muscular sarcocystosis develops in a biphasic pattern with initial fever and later prolonged myalgia, eosinophilia, and CK-elevation. Steroids achieve symptomatic relief in the late phase. Early cotrimoxazole-therapy could possibly prevent parasitic muscle invasion. In fever after travel to Malaysia differential diagnosis should include sarcocystosis. The source of infection appears to be located in North-West of Tioman. Further studies are needed, including addressing early diagnosis and treatment. © Georg Thieme

  17. Renal infection by a new coccidian genus in big brown bats (Eptesicus fuscus).

    Science.gov (United States)

    Wünschmann, Arno; Wellehan, James F X; Armien, Anibal; Bemrick, William J; Barnes, Donald; Averbeck, Gary A; Roback, Richard; Schwabenlander, Marc; D'Almeida, Edgar; Joki, Ron; Childress, April L; Cortinas, Roberto; Gardiner, Chris H; Greiner, Ellis C

    2010-02-01

    A novel coccidian parasite from the kidney of big brown bats (Eptesicus fuscus) is described. This coccidian (Nephroisospora eptesici nov. gen., n. sp.) was associated with a generally mild, focal or multifocal, well-demarcated cortical renal lesion less than 1 mm in diameter. The lesion represented cystic, dilated tubules with hypertrophied tubular epithelial cells and was present in the kidneys of 29 of 590 bats. Numerous coccidian parasites in various stages of development were present within the tubular epithelial cells and within the cyst lumina. Oocysts were collected from cystic dilated tubules. Thin-walled, sporulated ellipsoidal oocysts measuring an average of 18.9 x 20.8 microm were present in kidney tissue. The oocysts contained 2 sporocysts with 4 sporozoites. A polar body and a prominent oocyst residuum were present in the oocysts, but no micropyle, sporocyst residuum, or Stieda bodies were detected. Analysis of the 18S rRNA gene sequence put the parasite in the Sarcocystidae. The parasite is closely related to Besnoitia, Hammondia, Neospora, and Toxoplasma. Ultrastructural features, such as the presence of an apical complex in merozoites, support the identification of a coccidian. A new genus and species, Nephroisospora eptesicii, is proposed for this unusual coccidian in which the entire cycle is completed in the kidney of a single host; it has a membrane-like oocyst wall, sporogony occurs in the host rather than in the abiotic environment, and the positioning of the parasite by nucleic acid sequence indicates it to be closely allied to Sarcocystis and Besnoitia.

  18. Gastrointestinal parasites of stray cats in Kashan, Iran.

    Science.gov (United States)

    Mohsen, A; Hossein, H

    2009-04-01

    Considering the role of parasites in contamination of human beings and domestic animals and lack of information in the region, the present study was performed to investigate the infection status of helminthes and protozoa of stray cats in central Iran. A cross - sectional study was conducted on 113 stray cats trapped from different geographic regions of Kashan during four seasons and were necropsied. Different organs including: kidney, heart, liver, lungs, gastrointestinal tract and abdominal cavity were inspected for helminthes and protozoa infection. Animal's characters including: genus, weight, and season, location, microscopic and macroscopic findings were recorded in a special form. Data were classified and statistically analyzed with a confidence interval of 95%. Chi- Squire Test was used to show the relationship between different factors and parasitic infection. From a total 113 stray cats examined, 67(59.3%) were male and 46(40.7%) were female. Fifteen species of endoparasite including helminthes and protozoa were detected in intestine and fecal sample of the examined cats. There were six protozoa, five cestodes and four nematodes. All endoparasite were localized in the gastrointestinal tract. Overall 108 cats (95.6%) have been infected with at least one of the endoparasites. Prevalences of parasites found were Nematodea: Toxocara cati 13.3%, Physaloptera preputialis 39.8%, Rictularia 52.2% and Uncinaria stenocephala 1.8%; Cestodea: Mesocestoides lineatus 7.1%, Taenia taeniaformis 15%, Diplopylidium nolleri 64.6%, Dipylidium caninum 68.1% and Joyeuxiella echinorhyncoides 85%; Sporozoea: Isospora rivolta 5.3%, Isospora felis 5.3%, Sarcocystis spp 8%, Blastocystis spp 16.8% and Zoomastigophorea: Giardia felis 0.9% and Trichomonas spp 1.8%. Contamination rate for zoonotic parasites of cat was greater than expected in Kashan region. In this respect, appropriate control measures should be taken and it is recommended to determine the most appropriate preventive

  19. Health assessment of raptors in triage in Belo Horizonte, MG, Brazil

    Directory of Open Access Journals (Sweden)

    D de A Andery

    2013-09-01

    Full Text Available Falconiformes (n=82, Strigiformes (n=84 and Cathartiformes (n=14 at a triage center (CETAS-Belo Horizonte, IBAMA, Brazil were examined between 2008 and 2010 . No bird was reactive at hemagglutination-inhibition (HI for antibodies against Mycoplasma gallisepticum (Mg. Two Caracara plancus (2/68 had HI titers (16-32 against Newcastle disease virus. No Chlamydophila psittaci DNA was detected in the liver (PCR; n=95. Blood smears (Giemsa; n=89 and spleen fragments (PCR; n=82 were 13.5% and 8.5% positive, respectively, for Haemoproteus only. Necropsy of Cathartiformes (n=10, Falconiformes (n=42 and Strigiformes (n=57 showed that trauma injuries were the main cause (63.3% of admission and death, being fractures (38.5% of the thoracic limbs (57.1% the most frequent. Nematode (12.8%, cestode (1.8%, trematode (0.9%, and acanthocephalan (2.7% parasite infections were relevant. Mites (Acari were the most frequent (17.4% external parasites, particularly Ornithonyssus sylviarum in Asio clamator and Amblyomma cajennense in Tyto alba. Chewing lice (10.1% and Pseudolynchia spp. (9.2% were also found. Histomonas spp. (6.4% was found in the ceca of Bubo virginianus, Athene cunicularia, Tyto alba, and Asio clamator, but not in Falconiformes or Cathartiformes. Trichomonas spp. (oral cavity, pharynx and upper esophagus; 9.1% was detected in Falconiformes and Strigiformes, but not in Cathartiformes. Trichomonas spp. were found in A. cunicularia, Asio clamator, Glaucidium brasilianum and Tyto alba (Strigiformes, and in Rupornis magnirostris, Milvago chimachima, Falco femoralis, Falco sparverius and Caracara plancus (Falconiformes. Coccidia (9.1% (Sarcocystis spp., 6.4% and mycosis were observed in most Tyto alba (70%. The evaluated Orders may not pose risks for commercial poultry production. Habitat loss and urban adaptation may be increasingly affecting raptors.

  20. Aspectos ultraestruturais do processo de divisão do Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Wanderley de Souza

    1974-02-01

    Full Text Available Neste trabalho é feita uma revisão sobre alguns aspectos biológicos do Toxoplasma gondii, principalmente sobre a ultraestrutura da forma interfásica e as modificações ultraestruturais que ocorrem no parasito durante o seu processo de divisão. Considera-se inicialmente o processo de divisão binária admitindo-se, porém, a possibilidade de que as imagens interpretadas como senão de divisão binária representem estágios da divisão por endodiogenia. Quanto à endodiogenia descrevem-se as alterações que ocorrem na "parasito mãe" durante o processo de formação dos dois "parasitos filhos". Este processo é semelhante no Toxoplasma gondii, Besnoitia jellisoni, Sarcocystis tenella e Frenkelia. Discute-se a possibilidade da formação de mais de dois "parasitos filhos" por um processo de endopoligenia, bem como o processo de esquizogonia. Os resultados mais recentes mostram que não existe esquizogonia nas formas vsgetativas do Toxoplasma gondii, senão que as imagens interpretadas como tal, ao microscópio ótico, são o resultado de endodiogenias sucessivas em que os endozoitas formados permanecem ligados entre si pela região posterior. A esquizogonia é, no entanto, encontrada nas formas que se desenvolvem no interior de células epiteliais do intestino do gato, que é o hospedeiro definitivo do Toxoplasma gondii. Discute-se o conceito de esquizogonia, comparando-o em três protozoários: Eimeria bovis, E. callospermophili e Plasmodium juxtanucleare, que apresentam diferenças entre si quanto ao processo de iniciação da individualização dos "parasitos filhos". Refere-se à recente hipótese que considera a endodiogenia como o processo fundamental de divisão dos esporozoárlos, ocorrendo na fase final da esquizogonia. Finalmente é acentuado o papel que a microscopia eletrônica aliada às modernas técnicas de citoquímica e imunocitoquimica poderá desempenhar no sentido de um melhor conhecimento da biologia do Toxoplasma

  1. Structural and evolutionary adaptation of rhoptry kinases and pseudokinases, a family of coccidian virulence factors

    Science.gov (United States)

    2013-01-01

    Background The widespread protozoan parasite Toxoplasma gondii interferes with host cell functions by exporting the contents of a unique apical organelle, the rhoptry. Among the mix of secreted proteins are an expanded, lineage-specific family of protein kinases termed rhoptry kinases (ROPKs), several of which have been shown to be key virulence factors, including the pseudokinase ROP5. The extent and details of the diversification of this protein family are poorly understood. Results In this study, we comprehensively catalogued the ROPK family in the genomes of Toxoplasma gondii, Neospora caninum and Eimeria tenella, as well as portions of the unfinished genome of Sarcocystis neurona, and classified the identified genes into 42 distinct subfamilies. We systematically compared the rhoptry kinase protein sequences and structures to each other and to the broader superfamily of eukaryotic protein kinases to study the patterns of diversification and neofunctionalization in the ROPK family and its subfamilies. We identified three ROPK sub-clades of particular interest: those bearing a structurally conserved N-terminal extension to the kinase domain (NTE), an E. tenella-specific expansion, and a basal cluster including ROP35 and BPK1 that we term ROPKL. Structural analysis in light of the solved structures ROP2, ROP5, ROP8 and in comparison to typical eukaryotic protein kinases revealed ROPK-specific conservation patterns in two key regions of the kinase domain, surrounding a ROPK-conserved insert in the kinase hinge region and a disulfide bridge in the kinase substrate-binding lobe. We also examined conservation patterns specific to the NTE-bearing clade. We discuss the possible functional consequences of each. Conclusions Our work sheds light on several important but previously unrecognized features shared among rhoptry kinases, as well as the essential differences between active and degenerate protein kinases. We identify the most distinctive ROPK-specific features

  2. PREPARATION AND CHARACTERIZATION OF THE MONOCLONAL ANTIBODIES AGAINST THE EXCRETORY-SECRETORY ANTIGENS OF TRICHINELLA SPIRALIS MUSCLE LARVAE%旋毛虫肌幼虫ES Ag单克隆抗体的制备与特性鉴定

    Institute of Scientific and Technical Information of China (English)

    李学伍; 杨艳艳; 王自良; 肖治军; 柴书军; 张改平

    2008-01-01

    目的 获得能够用于制备旋毛虫病快速检测试纸条的ES Ag的单克隆抗体.方法 应用旋毛虫肌幼虫排泄分泌抗原(ES Ag)免疫诱导Balb/c小鼠,使小鼠产生较强的免疫应答,将免疫小鼠的脾细胞与NSO瘤细胞融合,利用ES-45、ES-49抗原通过间接ELISA对大量杂交瘤细胞培养上清的筛选,筛选出分泌高亲和力单克隆抗体的4株杂交瘤细胞.结果 ES-45和ES-49(分子质量分别为45ku,49ku)分别被Ts-2D4、Ts-4C5和Ts-4H6、Ts-2G8单克隆抗体识别,与猪肺丝虫(Metastrongylus pudendotectus.MP)、囊虫(Cysticercus cellulosae,CC)、细颈囊尾蚴(Cysticercus tenuicollis,CT)、蛔虫(Ascaris suum,AS)、弓形虫(Toxoplasma gondii,TG)、住肉孢子虫(Sarcocystis miescheriana,SM)抗原反应测试表明,4株单抗与参试抗原均无交叉反应,所有单抗上清ELISA平均效价为1∶1 120,腹水ELISA平均效价为1.1×106,亲和力常数的平均值为6.12 ×109L/mol.以金标免疫吸附试纸条原理为基础,利用制备的单抗成功研制了旋毛虫病快速检测试纸条,操作快速、无需设备及试剂,可以作为旋毛虫病的实时监测工具.结论 ES Ag单抗用于制备旋毛虫病快速诊断试纸条是理想的试剂.

  3. Multi-scale occupancy approach to estimate Toxoplasma gondii prevalence and detection probability in tissues: an application and guide for field sampling.

    Science.gov (United States)

    Elmore, Stacey A; Huyvaert, Kathryn P; Bailey, Larissa L; Iqbal, Asma; Su, Chunlei; Dixon, Brent R; Alisauskas, Ray T; Gajadhar, Alvin A; Jenkins, Emily J

    2016-08-01

    Increasingly, birds are recognised as important hosts for the ubiquitous parasite Toxoplasma gondii, although little experimental evidence exists to determine which tissues should be tested to maximise the detection probability of T. gondii. Also, Arctic-nesting geese are suspected to be important sources of T. gondii in terrestrial Arctic ecosystems, but the parasite has not previously been reported in the tissues of these geese. Using a domestic goose model, we applied a multi-scale occupancy framework to demonstrate that the probability of detection of T. gondii was highest in the brain (0.689, 95% confidence interval=0.486, 0.839) and the heart (0.809, 95% confidence interval=0.693, 0.888). Inoculated geese had an estimated T. gondii infection probability of 0.849, (95% confidence interval=0.643, 0.946), highlighting uncertainty in the system, even under experimental conditions. Guided by these results, we tested the brains and hearts of wild Ross's Geese (Chen rossii, n=50) and Lesser Snow Geese (Chen caerulescens, n=50) from Karrak Lake, Nunavut, Canada. We detected 51 suspected positive tissue samples from 33 wild geese using real-time PCR with melt-curve analysis. The wild goose prevalence estimates generated by our multi-scale occupancy analysis were higher than the naïve estimates of prevalence, indicating that multiple PCR repetitions on the same organs and testing more than one organ could improve T. gondii detection. Genetic characterisation revealed Type III T. gondii alleles in six wild geese and Sarcocystis spp. in 25 samples. Our study demonstrates that Arctic nesting geese are capable of harbouring T. gondii in their tissues and could transport the parasite from their southern overwintering grounds into the Arctic region. We demonstrate how a multi-scale occupancy framework can be used in a domestic animal model to guide resource-limited sample collection and tissue analysis in wildlife. Secondly, we confirm the value of traditional occupancy in

  4. Emerging food-borne parasites.

    Science.gov (United States)

    Dorny, P; Praet, N; Deckers, N; Gabriel, S

    2009-08-07

    Parasitic food-borne diseases are generally underrecognised, however they are becoming more common. Globalization of the food supply, increased international travel, increase of the population of highly susceptible persons, change in culinary habits, but also improved diagnostic tools and communication are some factors associated with the increased diagnosis of food-borne parasitic diseases worldwide. This paper reviews the most important emerging food-borne parasites, with emphasis on transmission routes. In a first part, waterborne parasites transmitted by contaminated food such as Cyclospora cayetanensis, Cryptosporidium and Giardia are discussed. Also human fasciolosis, of which the importance has only been recognised in the last decades, with total numbers of reported cases increasing from less than 3000 to 17 million, is looked at. Furthermore, fasciolopsiosis, an intestinal trematode of humans and pigs belongs to the waterborne parasites as well. A few parasites that may be transmitted through faecal contamination of foods and that have received renewed attention, such as Toxoplasma gondii, or that are (re-)emerging, such as Trypanosoma cruzi and Echinococcus spp., are briefly reviewed. In a second part, meat-borne parasite infections are reviewed. Humans get infected by eating raw or undercooked meat infected with cyst stages of these parasites. Meat inspection is the principal method applied in the control of Taenia spp. and Trichinella spp. However, it is often not very sensitive, frequently not practised, and not done for T. gondii and Sarcocystis spp. Meat of reptiles, amphibians and fish can be infected with a variety of parasites, including trematodes (Opisthorchis spp., Clonorchis sinensis, minute intestinal flukes), cestodes (Diphyllobothrium spp., Spirometra), nematodes (Gnathostoma, spp., anisakine parasites), and pentastomids that can cause zoonotic infections in humans when consumed raw or not properly cooked. Another important zoonotic food

  5. Food-borne pathogens of animal origin-diagnosis, prevention, control and their zoonotic significance: a review.

    Science.gov (United States)

    Dhama, K; Rajagunalan, S; Chakraborty, S; Verma, A K; Kumar, A; Tiwari, R; Kapoor, S

    2013-10-15

    The term food borne diseases or food-borne illnesses or more commonly food poisoning are used to denote gastrointestinal complications that occur following recent consumption of a particular food or drink. Millions of people suffer worldwide every year and the situation is quiet grave in developing nations creating social and economic strain. The food borne pathogens include various bacteria viz., Salmonella, Campylobacter, Escherichia coli, Listeria monocytogenes, Yersinia enterocolitica, Staphylococcus, Arcobacter, Clostridium perfringens, Cl. botulinum and Bacillus cereus and helminths viz., Taenia. They also include protozoa viz., Trichinella, Sarcocystis, Toxoplasma gondii and Cryptosporidium parvum. The zoonotic potential and the ability to elaborate toxins by many of the microbes causing fatal intoxication are sufficient to understand the seriousness of the situation. The viral agents being host specific their transmission to humans through food of animal origin is not yet confirmed although these animal viruses are similar to that of viruses infecting human. Food-borne bacteria; protozoa and helminthes have complex distribution pattern in the environment and inside the host system. This along with complexity of the maintenance chain and life cycle (of parasites) has made it difficult for epidemiologist and diagnostician to undertake any immediate safety measures against them. Serological and molecular diagnostic tests viz. ELISA, Latex agglutination test, Lateral flow assays, Immunomagnetic separation assays, molecular assays viz. Polymerase Chain Reaction (PCR), multiplex PCR, immuno-PCR, Realtime PCR, Random Amplified Polymorphic DNA (RAPD)-PCR, DNA microarrays and probes are widely used. Along with these LAMP assays, Capillary Electrophoresis-Single Strand Confirmation polymorphism (CE-SSCP); Flow cytometry, FISH, Biosensors, Direct epifluorescent filter technique, nanotechnology based methods and sophisticated tools (ultrasonography, magnetic resonance

  6. Aspectos ultraestruturais do processo de divisão do Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Wanderley de Souza

    1974-02-01

    Full Text Available Neste trabalho é feita uma revisão sobre alguns aspectos biológicos do Toxoplasma gondii, principalmente sobre a ultraestrutura da forma interfásica e as modificações ultraestruturais que ocorrem no parasito durante o seu processo de divisão. Considera-se inicialmente o processo de divisão binária admitindo-se, porém, a possibilidade de que as imagens interpretadas como senão de divisão binária representem estágios da divisão por endodiogenia. Quanto à endodiogenia descrevem-se as alterações que ocorrem na "parasito mãe" durante o processo de formação dos dois "parasitos filhos". Este processo é semelhante no Toxoplasma gondii, Besnoitia jellisoni, Sarcocystis tenella e Frenkelia. Discute-se a possibilidade da formação de mais de dois "parasitos filhos" por um processo de endopoligenia, bem como o processo de esquizogonia. Os resultados mais recentes mostram que não existe esquizogonia nas formas vsgetativas do Toxoplasma gondii, senão que as imagens interpretadas como tal, ao microscópio ótico, são o resultado de endodiogenias sucessivas em que os endozoitas formados permanecem ligados entre si pela região posterior. A esquizogonia é, no entanto, encontrada nas formas que se desenvolvem no interior de células epiteliais do intestino do gato, que é o hospedeiro definitivo do Toxoplasma gondii. Discute-se o conceito de esquizogonia, comparando-o em três protozoários: Eimeria bovis, E. callospermophili e Plasmodium juxtanucleare, que apresentam diferenças entre si quanto ao processo de iniciação da individualização dos "parasitos filhos". Refere-se à recente hipótese que considera a endodiogenia como o processo fundamental de divisão dos esporozoárlos, ocorrendo na fase final da esquizogonia. Finalmente é acentuado o papel que a microscopia eletrônica aliada às modernas técnicas de citoquímica e imunocitoquimica poderá desempenhar no sentido de um melhor conhecimento da biologia do Toxoplasma

  7. FTA-PCR检测水源性隐孢子虫研究%Detection of Cryptospordium spp.in environmental water samples by FTAPCR

    Institute of Scientific and Technical Information of China (English)

    张小萍; 朱倩; 何艳燕; 江莉; 蒋守富

    2011-01-01

    目的 建立FTA-PCR方法,并应用于水源中隐孢子虫污染的基因检测.方法 用免疫磁珠(IMS)分离纯化隐孢子卵囊,采用FTA卡提取隐孢子卵囊DNA,根据隐孢子虫18 S rRNA的基因序列设计引物,对模板DNA进行PCR扩增,同时以刚地弓形虫、猪人肉孢子虫、细粒棘球蚴和华支睾吸虫等DNA样本作对照.用琼脂糖电泳和溴乙腚染色对PCR产物进行检测.结果 微小隐孢子虫、安氏隐孢子虫和贝氏隐孢子虫均扩增出446 bp左右的DNA片段,其他对照DNA样本均未出现扩增反应.应用FTA-PCR检测上海市5个区自来水10份,未出现特异性条带;检测15份水源水、25份黄浦江水、15份动物饲养场周边河水、9份污水处理厂出水和6份生活污水排污口水,分别检测出0、2、5、1份和1份共9份阳性,阳性样品均扩增出约446 bp的特异性片段.结论 FTA-PCR可用于水样中隐孢子虫污染的基因检测.动物饲养场周边河水受隐孢子虫污染程度较高.%Objective To establish a FTA-polymeras chain reaction (FTA-PCR) method in detection of Cryptospordium spp.in different sources of water. Methods The semi automated immunomagnetic separation (IMS) of Cryptospordium oocysts in environmental water samples was performed firstly, and then genomic DNA of Cryptospordium oocysts was extracted by FTA filters disk. Oligonucleotide primers were designed based on the DNA fragment of the 18 S rRNA gene from C. parvum. Plate DNA was amplified with primers in PCR. The control DNA samples from Toxoplasma gondii,Sarcocystis suihominis, Echinococcus granulosus, and Clonorchis sinensis were amplified simultaneously. All PCR products were detected by agar electrophoresis dyed with ethidium bromide. Results The 446 bp fragment of DNA was detected in all samples of C. parvum, C. andersoni, and C. baileyi, while it was not detected in control groups in laboratory. No positive samples were found from 10 samples collected from tape water in 5

  8. Prevalence and risk factors of Giardia duodenalis in dogs from Romania.

    Science.gov (United States)

    Mircean, Viorica; Györke, Adriana; Cozma, Vasile

    2012-03-23

    in communities were identified as risk factors for infection by multivariate logistic regression analysis. 71.2% (37/52) Giardia cysts positive dogs presented co-infections with other intestinal parasites: Toxocara canis (14/52; 26.9%), Isospora ohioensis (12/52; 23.1%), Ancylostoma caninum (9/52; 17.3%), Uncinaria stenocephala (7/52; 13.5%), Trichocephalus vulpis (6/52; 11.5%), Hammondia heydorni/Neospora caninum (5/52; 9.6%), Sarcocystis spp. (5/52; 9.6%), Isospora canis (4/52; 7.7%), Capillaria aerophila (3/52; 5.8%), Strongyloides stercoralis (2/52; 93.8%), Dipylidium caninum (1/52; 1.9%) and Toxascaris leonina (1/52; 1.9%).

  9. Six new species of coccidia (Apicomplexa: Eimeriidae) from endangered Phelsuma spp. geckoes (Sauria: Gekkonidae) of the Black River Gorges National Park, Mauritius.

    Science.gov (United States)

    Daszak, Peter; Ball, Stanley J; Jones, Carl G; Streicker, Daniel G; Snow, Keith R

    2009-12-01

    of P. cepediana. These oocysts or sporocysts are significantly larger than the Cryptosporidium species so far described from reptiles, and likely represent excretion of spuriously ingested sporocysts of a Sarcocystis or Adelina coccidian.

  10. Intestinal Coccidia

    Directory of Open Access Journals (Sweden)

    MJ Ggaravi

    2007-06-01

    intestinal parasite is in most parts of the world. Sometimes it is noun traveller diarrhea Syndrom. The egg shapes of Oocyst are disporic tetrazoic. It is transmitted by vegetables and fruits. Trophozoite pass through schizogony step and repeats it several times. In the end of the cycle gametogony is done and the sexual forms will be repelled the human intestine. Symptoms are persistent diarrhoea, epigastric pain, headache, fever, vomitting and leanness, especially when physiologic disorder condition is seen in patient or they are in traveling. Misdiagnosis is a problem in laboratories but floatation method with zinc sulfate or sugar syrup is recommended. Sarcocystis: Sporocyst of S.hominis and S. suihominis is in the human feces, and the cyst form is in pig and cow muscles. It founds in tha tongue, pharynx and oesophagus muscles of habitant buffalo in Iran. Because of the large size of the cyst (1cm, it is seen with naked eyes and the risk of human infection is rare. If human eats raw or uncooked cow and pig meat, he will be infected with it. Sexual cycle is in the human body and sporocyst is repelled by the intestine. The disease may or may not have any symptoms. The symptoms are diarrhoea, stomach cramp, jejenuom and ileum necrosis. Diagnosis is based on concentrated floatation. The prevalence rate is too much in domestic animals.