In silico identification of immunotherapeutic and diagnostic targets in the glycosylphosphatidylinositol metabolism of the coccidian Sarcocystis aucheniae.

Meat of the South American camelids (SACs) llama and alpaca is an important source of animal protein and income for rural families in the Andes, and a product with significant growth potential for local and international markets. However, infestation with macroscopic cysts of the coccidian protozoon Sarcocystis aucheniae, a parasitosis known as SAC sarcocystosis, significantly hampers its commercialization. There are no validated methods to diagnose the presence of S. aucheniae cysts other than carcass examination. Moreover, there are no available drugs or vaccines to cure or prevent SAC sarcocystosis. Identification of relevant molecules that act at the host-pathogen interface can significantly contribute to the control of this disease. It has been shown for other pathogenic protozoa that glycosylphosphatidylinositol (GPI) is a critical molecule implicated in parasite survival and pathogenicity. This study focused on the identification of the enzymes that participate in the S. aucheniae GPI biosynthetic pathway and the repertoire of the parasite GPI-anchored proteins (GPI-APs). To this aim, RNA was extracted from parasite cysts and the transcriptome was sequenced and translated into amino acid sequences. The generated database was mined using sequences of well-characterized GPI biosynthetic enzymes of Saccharomyces cerevisiae and Toxoplasma gondii. Eleven enzymes predicted to participate in the S. aucheniae GPI biosynthetic pathway were identified. On the other hand, the database was searched for proteins carrying an N-terminal signal peptide and a single C-terminal transmembrane region containing a GPI anchor signal. Twenty-four GPI-anchored peptides were identified, of which nine are likely S. aucheniae-specific, and 15 are homologous to membrane proteins of other coccidians. Among the latter, 13 belong to the SRS domain superfamily, an extensive group of coccidian GPI-anchored proteins that mediate parasite interaction with their host. Phylogenetic analysis showed a great degree of intra- and inter-specific divergence among SRS family proteins. In vitro and in vivo experiments are needed to validate S. aucheniae GPI biosynthetic enzymes and GPI-APs as drug targets and/or as vaccine or diagnostic antigens.

Efecto del toltrazuril y la combinación de sulfadoxina y pirimetamina en el tratamiento de la sarcocistiosis canina durante el periodo patente / Effect of toltrazuril and the combination of sulfadoxine and pirimetamine in the treatment of canine sarcocystiosis during the prepatent period

El objetivo del estudio fue evaluar la eficacia del toltrazuril (2.5%) y de la combinación de sulfadoxina y pirimetamina (500/25 mg) en el tratamiento de la sarcocistiosis canina. El estudio se realizó durante los meses de octubre del 2005 a marzo del 2006, en la Estación Experimental del Centro de Investigaciones IVITA, Maranganí, Cusco, perteneciente a la Facultad de Medicina Veterinaria, UNMSM. Se utilizó 15 cachorros cruzados de ambos sexos, de 2-3 meses de edad, previamente desparasitados y alimentados solo con concentrado. Todos ellos fueron infectados con aproximadamente 9,000-10,000 quistes de Sarcocystis lamacanis, presentes en 4-5 g de tejido cardiaco de alpaca. Las heces se analizaron diariamente hasta observar la presencia de esporoquistes u ooquistes de Sarcocystis sp. Posteriormente fueron distribuidos en tres grupos de 5 animales c/u. Un grupo control no tratado y dos grupos tratados diariamente por 10 días, uno con toltrazuril (2.5%) en dosis de 15 mg/kpv y otro grupo con la combinación de sulfadoxina y pirimetamina (500/25 mg) en dosis de 20 y 1 mg/kgpv, respectivamente. Muestras fecales fueron recolectadas diariamente y se analizaron por el método de coproparasitológico de flotación con solución de Sheather para determinar la presencia de ooquistes o esporoquistes de Sarcocystis sp., y las muestras positivas se cuantificaron por el método de Stoll modificado, para determinar su número por gramo de heces. Los resultados muestran que el toltrazuril alcanzó una eficacia en el control de la sarcocistiosis del 94.7% al tercer día de tratamiento y un 100% al sexto día, mientras que la combinación de sulfadoxina y pirimetamina no logró controlarla, llegando a mostrar una moderada eficacia al segundo día post tratamiento (88.1%).

The objective of this study was to evaluate the efficacy of toltrazuril (2.5)% and the combination of sulfadoxine and pirimetamine (500-25 mg) on the treatment of canine sarcocystiosis. The study was conducted from October 2005 until March 2006 in the Experimental Station of the IVITA Research Center, Maranganí, Cusco. A total of 15 crossbred puppies, both sexes, 2-4 months of age, that were dewormed and fed with a diet extent of meat were used. Puppies were infected with approximately 9,000-10,000 cysts of Sarcosystis lamacanis present in 4-5 g of alpaca heart. Feces were daily collected and analyzed to observe the presence of sporocysts or oocysts. Then, were distributed in three groups of 5 animals each. One group remained untreated (as control) and the other two groups were daily treated during 10 days with 15 mg/kg of toltrazuril (2.5%) or the combination of sulfadoxine and pirimetamine (500-25 mg) in a dose of 20 and 1 mg/kg respectively. Fecal samples were daily collected and analyzed by the flotation method using the Sheather solution to determine the presence of sporocysts or oocysts and the modified Stoll method to quantify the number of eggs per gram of feces. The results showed that the efficacy of toltrazuril in the control of sarcosystiosis was 94.7% at the 3rd day of treatment and 100% at day 6, whereas the combination of sulfadoxine and pirimetamine did not control it, and only a moderate efficacy (88.1%) was observed in the 2nd day.

Evaluation of risk factors associated with clinical improvement and survival of horses with equine protozoal myeloencephalitis.

OBJECTIVE: To investigate risk factors for use in predicting clinical improvement and survival of horses with equine protozoal myeloencephalitis (EPM). DESIGN: Longitudinal epidemiologic study. ANIMALS: 251 horses with EPM. PROCEDURE: Between 1992 and 1995, 251 horses with EPM were admitted to our facility. A diagnosis of EPM was made on the basis of neurologic abnormalities and detection of antibody to Sarcocystis neurona or S neurona DNA in CSF. Data were obtained from hospital records and through telephone follow-up interviews. Factors associated with clinical improvement and survival were analyzed, using multivariable logistic regression. RESULTS: The likelihood of clinical improvement after diagnosis of EPM was lower in horses used for breeding and pleasure activities. Treatment for EPM increased the probability that a horse would have clinical improvement. The likelihood of survival among horses with EPM was lower among horses with more severe clinical signs and higher among horses that improved after EPM was diagnosed. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of horses with EPM is indicated in most situations; however, severity of clinical signs should be taken into consideration when making treatment decisions. Response to treatment is an important indicator of survival.

Gut Coccidia--Isospora, Cryptosporidium, Cyclospora and Sarcocystis.

The gut Coccidia are members of a large, varied, and exclusively intracellular group of protozoan parasites, four species of which (Isospora, Cryptosporidium, Cyclospora, and Sarcocystis) are human pathogens. The first three, but particularly Cryptosporidium parvum, have moved from medical curiosities to major problems with the coming of the acquired immunodeficiency syndrome (AIDS) epidemic, but are now known to also cause disease in the immunocompetent patient. They are easy to acquire and difficult to remove from the environment and, in the case of cryptosporidiosis, impossible to treat properly. Further research into many aspects of the biology of these organisms is urgently needed.