Protection studies with a globin-enriched protein fraction of Ostertagia ostertagi.

The protective capacity of an adult stage Ostertagia ostertagi globin antigen was tested in four vaccination experiments in cattle. In a preliminary experiment, calves were vaccinated three times intraperitoneally with 250 microg globin in Freund's adjuvant and challenged with a trickled infection of 25,000 infective larvae. In three subsequent field studies, calves were vaccinated twice or three times intramuscularly with 80-100 microg globin in Quil A and challenged with a natural gastrointestinal nematode infection on pasture. Higher globin-specific antibody levels were detected in the vaccinated calves than in the control animals in all vaccine trials. In the preliminary experiment, geometric mean cumulative egg counts in the globin group were reduced by 52% and total worm burdens were reduced by 28%, compared to the controls. In the first field trial cumulative faecal egg counts were reduced by 63% in the vaccinated calves. However, the reduction in faecal egg output in these two experiments was not statistically significant and no reduction in faecal egg counts was observed in the vaccinated animals in the two last field trials. In conclusion, vaccination of calves with O. ostertagi globin resulted in highly variable protection levels after challenge infection.

Inverse expression of peroxisomes and connexin-43 in the granulosa cells of the quail follicle.

Studying the regulation of peroxisome (Px) expression could improve our understanding of human peroxisomal disorders. The granulosa of the largest preovulatory quail follicles proved to be a relevant model because (a) Px expression changes according to the follicular maturation stage and (b) Px expression varies regionally according to the distance of the granulosa relative to the germinal disc region containing the female gamete (oocyte). The question was asked whether Px expression is related to the extent of metabolic cell coupling and whether zonal Px variation is causally related to oocytal factors. This was evaluated by the presence of catalase and Cx-43 (marker proteins for peroxisomes and gap junctions, respectively) and by in vitro experiments with granulosa explants. The data obtained show that the expression of Cx-43 and Px is inversely correlated both temporally and spatially. Uncoupling of gap junctions results in an upregulation of alpha-catalase immunofluorescence. This is in agreement with reports that gap junctions are often negatively affected by Px proliferators. The zonal gradient in Px expression appears to be imposed by the oocyte, as is the case for steroidogenesis and proliferative capacity in the granulosa epithelium. (J Histochem Cytochem 48:167-177, 2000)

Pathophysiological aspects of Mecistocirrus digitatus (Nematoda: Trichostrongylidae) infection in calves.

Three groups of four calves were experimentally infected with infectious larvae of Mecistocirrus digitatus. One group received a trickle infection of 5000 L3 per day for 8 days, the other two groups received a single infection of 5000 and 40,000 L3, respectively. All animals were necropsied 120 days after infection. Prepatent periods varied between 61 and 79 days, and maximal faecal egg output was reached between 80 and 100 days after infection. Repeated infections were not additive and worm counts at 120 days after infection varied between 100 and 440 in the first two groups and between 120 and 1700 in the last group. There was no significant difference in worm counts between the different infection regimes. However, there was a significant positive relationship between worm burden and faecal egg counts. In addition, there were significant negative relationships between worm burden and packed cell volume, and weight gain. The relative decreases in packed cell volume and weight gain emerged 70-80 days after infection. Serum pepsinogen levels were significantly elevated by the end of the trial, but the observed positive relationship between worm burden and pepsinogen was not significant. An enzyme immunoassay based on crude adult antigen was able to detect M. digitatus infection at 90 and 100 days after infection, but again there was no significant association between worm burden and antibody levels. Therefore, anaemia and a reduction in weight gain caused by the haematophagous activity of adult stages seem to be the most important pathogenic effects of M. digitatus infection in calves.

Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of sarcoptic mange in swine.

An enzyme-linked immunosorbent assay (ELISA) using an extract from Sarcoptes scabiei var. culpes as antigen was evaluated for its usefulness in monitoring antibodies to S. scabiei var. suis in naturally infected pigs. Five hundred and fifty-eight serum samples from certified Sarcoptes-free pigs (120 from weaned piglets, 218 from fatteners and 220 from adult sows) and 94 samples from certified Sarcoptes-infected pigs (35 from weaned piglets, 45 from fatteners and 14 from adult sows) were examined. The cut-off optical density (OD) values, considered to reflect the presence of antibodies to S. scabiei var. suis, were determined to be 0.059, 0.213 and 0.374 for weaners, fatteners and sows, respectively. The specificity of the ELISA was high (> or = 98%) for all age groups. Sensitivity was highest in the piglets (80%), followed by fatteners (78%) and sows (50%). These results indicate that an ELISA may be a simple and valuable alternative to monitor S. seabiei var. suis infections in naturally infected weaners and fatteners.

Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults.

Western blot analysis using an anti-globin rabbit serum Rb94 revealed a major band of 17 kDa in extracts of Ostertagia ostertagi adults and 4th-stage larvae. The adult stage globin-like antigen (OoAdGlb) was purified from total worm extracts by liquid chromatography. The protein has an estimated molecular mass of 36 kDa under non-reducing conditions, suggesting a dimeric structure containing 2 non-covalently linked 17 kDa monomers. Tryptic peptides were sequenced and showed strong similarities with the globins of free-living and parasitic nematodes. Immunolocalization revealed the presence of this globin-like antigen in the body wall musculature and/or the cuticle of O. ostertagi adults. An enzyme-linked immunosorbent assay based on the purified OoAdGlb showed no differences in response between calves infected by O. ostertagi and/or Cooperia oncophora and the negative controls.