G. Savini

The epidemiology of Sarcocystis spp. in cattle of Western Australia

Oesophagus samples from 714 cattle from Western Australia were examined by artificial digestion to detect the presence of Sarcocystis spp. The overall prevalence of infection was 52%. The prevalence of infection increased with age and was highest in the entire males (92%). The prevalence was lower in cattle which originated from arid and semiarid regions (9 and 31% respectively) than those from tropical (87%) and temperate (60%) regions. Possible reasons for these differences are discussed and it is concluded that environmental and management factors as well as host age and sex influence the prevalence of infection with Sarcocystis spp. in cattle.

Viability of the sporocysts of Sarcocystis cruzi after exposure to different temperatures and relative humidities

The effect of temperature and relative humidity (RH) on the survival of sporocysts of S. cruzi were investigated in vitro. Under all experimental conditions (temperature of 4 degrees C, 37 degrees C, or room temperature; RH of 18%, 75%, or 100%) some sporozoites retained their viability to excyst for at least 90 days. The best conditions for survival were 4 degrees C at 100% RH (more than 240 days) and 37 degrees C at 18% RH (more than 180 days). Sporocysts maintained at room temperature at all humidities had the lowest level of survival. It is concluded that sporocysts of S. cruzi are able to survive in most environments for several months and that the fluctuation of the daily ambient temperature is likely to influence the viability of the sporocysts.

STUDIES ON PATHOGENESIS, TISSUE INFECTION AND CONGENITAL TRANSMISSION IN COWS EXPERIMENTALLY INFECTED WITH SARCOCYSTIS CRUZI BY VARIOUS ROUTES

A group of nine cows, naturally infected with Sarcocystis, were challenged with Sarcocystis cruzi: three intrarumenally with sporocysts, two intrarumenally with water (controls), two intravenously with merozoites grown in vitro and two intravenously with saline solution (controls). The animals intrarumenally challenged with sporocysts developed acute sarcocystiosis and produced stillborn calves, whereas those intravenously challenged with merozoites suffered from subclinical sarcocystiosis with premature births. No parasites were found in calves from cows challenged with sporocysts; however, a meront of Sarcocystis was found within a macrophage in the cerebrospinal fluid of a calf from a cow intravenously inoculated with merozoites of S. cruzi. This is the first time that merozoites of S. cruzi grown in vitro have been demonstrated to retain the ability to infect their natural intermediate host and complete their life cycle.

Excystation rates and infectivity of sporocysts of Sarcocystis cruzi exposed to different treatments and storages

The effect of some chemical and anatomical factors on the excystability and infectivity in cell culture of sporocysts of Sarcocystis cruzi was investigated. A significantly (P < 0.001) higher excystation rate (ER) occurred when sodium bicarbonate was added to the excysting fluid at concentrations between 0.075 and 0.3 M. Sporocysts collected from different dogs had different ER. In contrast, although not uniformly distributed along the intestinal lumen, the sporocysts collected from the different tracts of the intestine showed similar ER. The period of storage did not affect the excystability of the sporocysts; however, it influenced the pattern of growth of the sporozoites in cell culture. Sporozoites excysted from sporocysts stored for approximately 2 years grew slower and produced significantly fewer merozoites compared to those excysted from sporocysts stored for 5-7 months.

Class-specific antibody responses in cattle following experimental challenge with sporocysts or merozoites of Sarcocystis cruzi.

An ELISA using antigen produced from merozoites of Sarcocystis cruzi was developed to monitor specific IgM and IgG antibody, following challenge of cattle with either merozoites or sporocysts of S. cruzi. This assay was compared with an ELISA using antigen produced from the cystozoite stage of the parasite. Both ELISAs were able to detect significant increases in levels of circulating IgM and IgG antibodies against Sarcocystis in all challenged cows; however, the magnitude of the titres was greater in the ELISA which used the antigen derived from the merozoites. This immunoassay also detected increases in the levels of IgG earlier than did the assay using antigen derived from cystozoites of S. cruzi. Since this rise coincided with the presence of clinical signs, and was persistent for several weeks, the IgG-ELISA using antigen derived from merozoites appears to be suitable for the diagnosis of acute sarcocystiosis in cattle. Furthermore, since significant increases in the levels of circulating IgM and IgG antibodies against Sarcocystis were detected in the cows infected with merozoites of S. cruzi, it is evident that merozoites of S. cruzi cultured in vitro maintain their capability to replicate in the natural intermediate host.

Sensitivities and specificities of two ELISA tests for detecting infection with Sarcocystis in cattle of Western Australia

The accuracies of two ELISAs, one using antigen from merozoites of S. cruzi grown in vitro and the other using antigen from cystozoites of S. cruzi, for detecting infection of cattle with Sarcocystis, were evaluated by testing the sera of 303 cattle from 36 Western Australian herds. The results were compared with those obtained by digestion of oesophageal samples collected from the same animals. A similar proportion of infected animals were detected by the three methods. The sensitivity of the assays for detecting infected cattle was comparable (98 and 95% for the assay using antigen from merozoites and cystozoites, respectively), however the specificity (97%) of the assay which used antigen derived from merozoites was significantly higher (P < 0.001) than that (84%) which used antigen from cystozoites. When herds which had at least five animals sampled were considered, the same infected and non-infected herds were detected by the ELISA employing antigen from merozoites and the digestion methods (sensitivity and specificity of 100%). The sensitivity and specificity of the assay using cystozoite antigen were 100 and 67%, respectively. The kappa values for agreement beyond chance between the two ELISAs were calculated as 78% for the animal-based data and 72.5% for the herd-based data. We conclude that because of the high sensitivity and specificity of the ELISA using antigen derived from merozoites, this assay would be a useful and reliable tool for general sero-epidemiological studies into infection with Sarcocystis.

Risk factors associated with the occurence of sarcocystosis in Western Australia: results of a postal survey

A postal survey was conducted to identify important risk factors associated with the presence or absence of bovine sarcocystosis in a sample of 127 Western Australian cattle farmers. Seventy-three replies were received and analysed using the odds ratio (OR) method. A smaller proportion (P<0.0001) of herds were infected in the northern area (31.2%) than in the southern area (84.2%) and a significantly (P<0.025) higher stocking rate was found in herds infected by Sarcocystis (View the MathML source) than in non-infected herds (View the MathML source). Herds infected by Sarcocystis spp. had higher odds (OR=5.4) (P<0.05) of reporting one or more cases of bovine abortion than non-infected herds. Positive associations were found between infected herds and dogs fed raw meat, the presence of working dogs and the practice of leaving carcases in the field. Foxes were found to be strongly associated (OR=9.17, P<0.01) with Sarcocystis infection in those herds where carcases were not properly disposed of, whereas dingoes and feral dogs (OR=0.20) and feral cats (OR=0.32) were found to be associated with low odds of Sarcocystis infection.