Structure of Ostertagia ostertagi ASP-1: insights into disulfide-mediated cyclization and dimerization.

The cysteine-rich secretory/antigen 5/pathogenesis-related 1 (CAP) protein superfamily is composed of a functionally diverse group of members that are found in both eukaryotes and prokaryotes. The excretome/secretome of numerous helminths (parasitic nematodes) contains abundant amounts of CAP members termed activation-associated secreted proteins (ASPs). Although ASPs are necessary for the parasitic life cycle in the host, the current lack of structural and functional information limits both understanding of their actual role in host-parasite interactions and the development of new routes in controlling parasitic infections and diseases. Alleviating this knowledge gap, a 1.85 Å resolution structure of recombinantly produced Oo-ASP-1 from Ostertagia ostertagi, which is one of the most prevalent gastrointestinal parasites in cattle worldwide, was solved. Overall, Oo-ASP-1 displays the common hallmark architecture shared by all CAP-superfamily members, including the N-terminal CAP and C-terminal cysteine-rich domains, but it also reveals a number of highly peculiar features. In agreement with studies of the natively produced protein, the crystal structure shows that Oo-ASP-1 forms a stable dimer that has been found to be primarily maintained via an intermolecular disulfide bridge, hence the small interaction surface of only 306.8 Å(2). Moreover, unlike any other ASP described to date, an additional intramolecular disulfide bridge links the N- and C-termini of each monomer, thereby yielding a quasi-cyclic molecule. Taken together, the insights presented here form an initial step towards a better understanding of the actual biological role(s) that this ASP plays in host-parasite interactions. The structure is also essential to help to define the key regions of the protein suitable for development of ASP-based vaccines, which would enable the current issues surrounding anthelmintic resistance in the treatment of parasitic infections and diseases to be circumvented.

Development and distribution of bovine trichostrongyle infective larvae on a pasture irrigated by flooding or by spraying.

To compare the ecological effects of irrigation of a pasture, by flooding or by spraying, on the gastro-intestinal parasites of calves, two contiguous plots, one flooded and the other sprayed, were grazed for 10 days by naturally infected animals. Faeces, grass and soil under pats were sampled for 17-20 weeks for larval counts. The experiment was performed once in the spring and once in the summer. In the spring the maximal development rates in sprayed and flooded plots were 7.8 and 12.6 stage 3 larvae (L3)/100 eggs, respectively, for Cooperia and 3.5 and 3.2 L3/100 eggs, respectively for Ostertagia. During the summer, development rates were less than 1%. The main differences were found in the vertical distribution of the larvae: spraying favoured migration on grass and flooding favoured migration in soil, this finding being similar for both genera. The spatial distribution, studied by dividing each plot into 18 subplots, was a little more regular on grass in the sprayed plot. We conclude that the infection risk for calves is lower in flooded pasture than in sprayed pasture.

Ostertagia ostertagi in neonatal calves: establishment of infection in ruminating and non-ruminating calves.

An experiment was carried out to study the role of the ruminal function in the establishment of Ostertagia ostertagi in neonatal calves. Three groups of calves were fed either milk only (groups A and C), or hay and concentrate in addition to milk (group B) from birth. At the time of infection, ruminal function was negligible in groups A and C, whereas it was well developed in group B. Calves of groups A and B were each given 25,000 normal ensheathed infective larvae of O ostertagi and those of group C were given 25,000 infective larvae exsheathed in vitro. Daily faecal egg output and post mortem worm counts 28 days after infection were higher in calves with well developed ruminal function than those having only negligible ruminal function. In the latter group, exsheathed larvae established at a lower rate than did ensheathed larvae. The results suggest that the degree of development of the ruminal function influences the establishment of O ostertagi.

The influence of a low cobalt intake on the neutrophil function and severity of Ostertagia infection in cattle.

Two trials involving housed cattle examined the effect of Co depletion and supplementation on immune status as assessed by the neutrophil function test which measures the ability of isolated neutrophils to kill the yeast Candida albicans. A third trial investigated the extent to which Co status influenced the severity of Ostertagia ostertagi infection. In the first two trials liveweight gains were unaffected until some 40-60 weeks on the low dietary Co intake despite very low serum vitamin B12 values being recorded after 10 weeks. However, the immune status as measured by the neutrophil function test was reduced within 10 weeks of commencing the low Co diet. On administration of Ostertagia ostertagi larvae, Co-depleted cattle showed a greater weight loss than Co-supplemented cattle but showed no difference in the length of the prepatent period, worm egg production or serum gastrin and pepsinogen concentrations. After anthelmintic treatment both groups showed a similar response. It is postulated that the lowered immune response of Co-depleted cattle resulted in the greater severity of the Ostertagia ostertagi infection.

Efficacy of ivermectin against benzimidazole-resistant nematodes of sheep.

Two trials involving a total of 36 Dorset horn lambs were conducted to assess the anthelmintic efficacy of ivermectin against experimental infections of benzimidazole-resistant strains of Haemonchus contortus and Ostertagia circumcincta. Two resistant strains of each of the two species were used and in each trial the lambs were allocated to three groups. One group was given 200 micrograms ivermectin/kg bodyweight orally, the second group was given 5 mg oxfendazole/kg bodyweight orally and the third group remained untreated as controls. Fourteen days after treatment the lambs were necropsied. Ivermectin was found to be more than 99 per cent to 100 per cent effective against all four benzimidazole-resistant strains, whereas oxfendazole was 78.6 per cent and 83.8 per cent effective against the H contortus strains, and 25.6 per cent and 39.8 per cent effective against the O circumcincta strains.

Adverse immune reactions and the pathogenesis of Ostertagia ostertagi infections in calves.

The possible development of type-1 hypersensitivity reactions in the abomasal mucosa caused by soluble L3 products of Ostertagia ostertagi was studied in 4-month-old calves sensitized by repeated exposure to L3 over a 50-day period followed by anthelmintic treatment. Four groups each of 4 calves were used. Group 1 served as nonsensitized controls and group 2 as sensitized controls, group 3 was challenge exposed at 2-week intervals beginning at week 10 with a soluble L3 product (OAG), and group 4 was challenge exposed at 2-week intervals with an oral dose of L3, followed by anthelmintic treatment 3 days later. All calves infected with L3 became sensitized, as indicated by a positive reaction to an intradermal skin test. However, a passive cutaneous anaphylaxis was only partly effective in indicating the presence of homocytotropic antibody in the infected calves. Sensitized calves had significantly (P less than 0.05) higher eosinophil counts and plasma pepsinogen values for the entire 14 weeks than uninfected controls. Globule leukocyte and mast cell counts from the abomasal mucosa were also significantly (P less than 0.05) higher. Studies for possible immunomodulation revealed that lymphocyte counts decreased between every 2-week challenge-exposure period for groups-3 and -4 calves. A transient depression of blood lymphocyte (BL) responses to phytohemagglutinin (PHA), a T-cell mitogen, was observed over the first 8 weeks in the infected calves. Increases in BL responses to OAG were also observed. Differences were not observed in BL responses to pokeweed mitogen, a T- and B-cell mitogen. Blood lymphocyte responses to PHA in group-3 calves were low following the initial challenge exposure with OAG.(ABSTRACT TRUNCATED AT 250 WORDS)

Pathogenesis of simulated natural infections with Ostertagia ostertagi in calves.

The possibility of a mucosal hypersensitivity reaction and its relationship to the pathogenesis of simulated natural infections with Ostertagia ostertagi were studied in calves. Four groups of 4 calves each were used. One group was used as noninfected control; a 2nd group was given increasing doses of infective larvae; a 3rd group was given increasing doses of larvae and these were removed by succeeding treatment with an anthelmintic; and a 4th group was given an initial dose of larvae which was then eliminated with an anthelmintic. All calves given larvae became sensitized, as shown by an intradermal skin test. The continuously infected calves had significantly (P less than 0.05) higher fecal egg counts, eosinophil counts, plasma pepsinogen values, and worm burdens and significantly (P less than 0.05) lower lymphocyte counts than did the other groups of calves. These animals also had the most extensive mucosal pathologic changes. The group given intermittent larval challenge exposures followed by an anthelmintic showed decreased lymphocyte values, but these were not significant. Plasma pepsinogen values of this group increased between every challenge exposure and treatment, a 3-day period. This indicated that a mucosal hypersensitivity reaction had occurred in these calves at these times, because they were shown to have been sensitized, and challenge-exposure infections were not present for sufficient time to have produced direct pathologic effects. It therefore seems that a part of the pathologic changes in O ostertagi infections may be the result of the continuous challenge exposure experienced by the animals through a constant intake of larvae from pasture and the intestinal reaction to this challenge exposure.