Sarcocystis camelicanis increases interleukin (IL)-6 expression in one-humped camels (Camelus dromedarius) from Riyadh and Al Qassim, Saudi Arabia.

Sarcocystis spp. are intracellular protozoan parasites with an intermediate-definitive host life cycle based on a prey-predator relationship. Sarcocystis infection is common among different vertebrates including humans. The pathogenicity of Sarcocystis spp. is of varied significance including a possible lethal effect for the host. The goal of the present study was to investigate the inflammatory activity of Sarcocystis spp. in different organs of naturally infected camels. The tongue, esophagus, heart, diaphragm, and skeletal muscles were collected from 50 camels, and the tissues assessed for the presence of Sarcocystis spp. by macroscopic examination, light microscopy, and transmission electron microscopy (TEM). Moreover, expression of the interleukin (IL)-6 was analyzed using reverse transcriptase quantitative polymerase chain reaction (qPCR). Microscopic Sarcocystis spp. cysts were found in camels. TEM identified the cysts as Sarcocystis camelicanis (S. camelicanis). Sarcocystis infection increased inflammation by stimulation of IL-6 expression in different organs of the camels, particularly in those from the Al-Qassim region.

Oxidative stress and hepatic injury induced in mice fed a Sarcocystis hirsuta cyst extract.

We studied the toxic effects of a Sarcocystis hirsuta cyst extract fed to mice. Degenerative changes were found in mice gavage-fed fresh, frozen, and heat-treated S. hirsuta cyst extract. There were increases in the levels of serum aspartate aminotransferase and alanine aminotransferase as well as hepatic and brain malondialdehyde (MDA) levels along with concomitant decreases in catalase (CAT) and superoxide dismutase (SOD) activities of mice receiving fresh and frozen S. hirsuta extracts. Gavage feeding of heat-treated S. hirsuta cyst extract had no effects on liver enzymes or brain MDA content, but the liver MDA level did increase. Mice in the heat-treated cyst group showed reduced CAT and SOD activities as well as increased hepatic MDA levels compared to those in the control group. These results indicate that an extract of S. hirsuta cyst can induce oxidative stress and hepatic injury, even after heat treatment.

PAN-modular structure of microneme protein SML-2 from the parasite Sarcocystis muris at 1.95 Å resolution and its complex with 1-thio-ß-D-galactose.

The microneme protein SML-2 is a member of a small family of galactose-specific lectins that play a role during host-cell invasion by the apicomplexan parasite Sarcocystis muris. The structures of apo SML-2 and the 1-thio-ß-D-galactose-SML-2 complex were determined at 1.95 and 2.1 Å resolution, respectively, by sulfur-SAD phasing. Highly elongated dimers are formed by PAN-domain tandems in the protomer, bearing the galactose-binding cavities at the distal apple-like domains. The detailed structure of the binding site in SML-2 explains the high specificity of galactose-endgroup binding and the broader specificity of the related Toxoplasma gondii protein TgMIC4 towards galactose and glucose. A large buried surface of highly hydrophobic character and 24 intersubunit hydrogen bonds stabilize the dimers and half of the 12 disulfides per dimer are shielded from the solvent by the polypeptide chain, thereby enhancing the resistance of the parasite protein towards unfolding and proteolysis that allows it to survive within the intestinal tracts of the intermediate and final hosts.

Protozoal meningoencephalitis in sea otters (Enhydra lutris): a histopathological and immunohistochemical study of naturally occurring cases.

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.

Reduced levels of nitric oxide metabolites in cerebrospinal fluid are associated with equine protozoal myeloencephalitis.

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 (NO(x)(-)) in the cerebrospinal fluid (CSF) is correlated with the development of EPM. CSF NO(x)(-) 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 NO(x)(-) 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 NO(x)(-) 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 NO(x)(-) 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 NO(x)(-) concentration than horses that did not have neurologic signs (n = 24). In conclusion, lower CSF NO(x)(-) levels were associated with clinical EPM, suggesting that measurement of CSF NO(x)(-) 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 causally related to the development of EPM.

Lipid metabolism and Sarcocystis miescheriana infection in growing swine.

Sixteen 2-month-old pigs were divided into four equal groups and infected with either 500,000, 1,000,000 or 3,000,000 sporocysts of Sarcocystis miescheriana. Four pigs served as uninfected controls. Pigs were bled weekly and serum was collected beginning 14 days prior to infection and continuing until 63 days after infection. Body fat composition, as measured by the specific gravity of the carcass, was not affected by infection. There were no significant effects of infection on serum concentrations of glucose, insulin, triglycerides, and total, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) cholesterol. A slight depression in HDL cholesterol occurred during the acute phase of infection. Tumor necrosis factor (TNF) was not detected in serum from infected swine when assayed by a cytotoxicity assay using TNF-sensitive WEHI 164 clone 13 cells. Attempts to stimulate TNF production in RAW 264.7 cells with parasitic lysates gave mixed results. This study suggests that the disruption of lipid metabolism is not the primary cause of growth retardation in growing swine infected with S. miescheriana.

[The cytochemical study of polysaccharides in coccidia of the genus Sarcocystis. I. Sulfated glycosaminoglycans in Sarcocystis muris sarcocysts]. / Tsitokhimicheskoe issledovanie polisakharidov u koktsidii roda Sarcocystis. I. Sul'fatirovannye glikozaminoglikany v sarkotsistakh Sarcosystis muris.

An electron microscope study of sulfatized glycosaminoglycans (SG) was made for cyst stages of S. muris. The polysaccharides were detected in the submembranous and subwall layers of the sarcocysts, in addition to the ground substance and septae. Moreover SG were discovered in the cyst stages themselves--metrocytes, intermediate cells and merozoites (gamonts). SG were discernible as electron dark spots in vacuoles of the metrocytes. SG shaped as granules were scattered in the cytoplasm of both intermediate cells and merozoites. More granules of SG were seen in the cytoplasm of the merozoites compared to the intermediate cells. Thus, the quantity, localization and structure of SG are seen to follow the process of differentiation in muscle cysts of S. muris.

Plasma and tissue concentrations and molecular forms of somatostatin in calves infected with Sarcocystis cruzi.

The effects of parasitic infection on plasma and tissue content of immunoreactive somatostatin (SRIF) were studied in 4-mo old male calves inoculated with the protozoan Sarcocystis cruzi. Because feed intake significantly decreased (70%) in infected calves around day 28 postinfection (pi), concomitant with the asexual replication of S. cruzi and outward expression of clinical signs, the relative contributions of infection and associated reduction in nutrition on plasma SRIF were evaluated. Treatment groups were: noninfected ad libitum fed (C), infected (250,000 S. cruzi oocysts per os) ad libitum fed (I) and noninfected calves pairfed to the level of intake of each infected calf (PF). Mean plasma concentrations of SRIF (pg/ml) on day 30 pi were: C, 224 +/- 22; I, 742 +/- 150; PF, 246 +/- 31 (effect of infection P less than .05). In another study, SRIF was measured in plasma and in pancreatic, duodenal, jejunal and ileal tissue extracts from normal and S. cruzi infected calves. Plasma and tissue samples were collected on day 42 pi. Mean plasma SRIF were 2.5 times higher in infected than control calves. Plasma insulin and insulin-like growth factor 1 was lower in infected v control calves (P less than .02). Plasma glucagon was similar between groups. Duodenal (P less than .05) and jejunal (P less than .02) SRIF content was higher in infected than control calves. Chromatography of tissue extracts on Sephadex G-50 revealed that the increase in SRIF was accounted for, in part, by molecular forms larger than cyclic SRIF-14. Data suggest that peripheral SRIF is increased in calves during S. cruzi infection. The increase in SRIF is not solely related to plane of nutrition. Altered levels of gut SRIF(s) may be associated with perturbed metabolic regulation in parasitized animals through direct effects on the gut.

Effects of Sarcocystis miescheriana infection on carcass quality and on the water-binding capacity of the meat of halothane-tested fattening pigs.

Ten halothane-positive pigs (stress sensitive, group A) and ten halothane-negative pigs (stress insensitive, Group C) with a mean body weight of 36 kg were each inoculated orally with 50,000 sporocysts of Sarcocystis miescheriana. Twelve halthane-positive pigs (Group B) and ten halothane-negative pigs (Group D) served as non-infected controls. Thirteen weeks post infection (p.i.) the lean: fat ratios of the pigs of the infected groups A and C were lower (A, 1:0.41 +/- 0.09; C, 1:0.50 +/- 0.10) than those of the pigs of the non-infected groups B and D (B, 1:0.50 +/- 0.08; D, 1:0.55 +/- 0.08). The back-fat thickness, the fat thickness 'A' and the fat thickness 'B' were thinner in infected pigs than in non-infected pigs. The difference in Lendenstärkespeckquotient (Loin Fat Thickness Quotient) (LSQ) between infected and non-infected pigs was not statistically significant. The values of the water-holding capacity were lower in infected pigs than in non-infected pigs, the difference being statistically significant only in the halothane-negative groups (C, 0.45 +/- 0.02; D, 0.48 +/- 0.04). The water-absorbing capacity was significantly higher in the infected groups (A, 5.92 +/- 3.99%; B, 2.26 +/- 1.08%; C, 8.96 +/- 2.90%; D, 4.97 +/- 2.51%). In conclusion, it can be said that there was a slight tendency towards a better carcass quality and a better water-binding capacity in infected pigs, although this was combined with reduced growth rates.

The effects of Sarcocystis miescheriana infections on blood enzymes and weight gain of stress-sensitive and stress-insensitive pigs.

The effects of mild Sarcocystis miescheriana infection on blood enzymes and body weight were compared in stress-sensitive (halothane positive) and in stress-insensitive (halothane negative) pigs. Ten halothane-positive pigs (Group A) and ten halothane-negative pigs (Group C) with a mean body weight of 36 kg were each inoculated orally with 50,000 sporocysts of S. miescheriana. Twelve halothane-positive pigs (Group B) and ten halothane-negative pigs (Group D) served as non-infected controls. Five days before infection (a.i.) and 58 days post infection (p.i.) all pigs were myostress challenged (creatine kinase test). During the whole period of the experiment (3 weeks a.i. to 13 weeks p.i.) body weights were determined and blood samples taken at weekly intervals. Creatine kinase (CK), aspartate aminotransferase (ASAT) and Sarcocystis antibody titres were determined. The infection induced slightly elevated temperatures (max. 42.8 degrees C) and transient reduced food intake during the 2nd and 3rd week p.i. The CK values of the infected pigs in Groups A and C increased from Day 28 p.i. onwards, and were significantly higher than those of the pigs of the non-infected controls (Days 35-77 p.i.). The ASAT values of the infected groups (A and C) increased from Day 21 p.i. onwards, and were significantly higher than those of the non-infected controls from Days 28-77 p.i. The myostress injection at 5 days a.i. (1st CK test) resulted in significantly higher CK and ASAT values in stress-sensitive pigs.(ABSTRACT TRUNCATED AT 250 WORDS)