SUMMARY Previous experiments have shown that genetic resistance to infection by Teladorsagia circumcincta in sheep can be measured by the level of IgA in gastric mucus, jointly with other parameters. The aim of this study has been to observe the influence of IgA on adult worms. The experiment was carried out with Churra sheep experimentally infected with T. circumcincta. At slaughter, gastric content, gastric mucus, blood samples and faeces were recovered to determine the number of eggs in utero, length of adult females, worm burden, number of L4, titre of serum pepsinogen, peripheral eosinophilia and eggs per gram (epg). IgA activity in gastric mucus, serum, nasal secretions and saliva were tested against somatic antigen from fourth-stage larvae (L4), somatic antigen from the adult stage and excretory-secretory (E/S) antigen from the adult stage. The results showed a significant correlation between serum IgA and gastric mucus (P<0.01) as well as in nasal secretions (P<0.01). We found negative correlations between IgA activity in gastric mucus with the eggs in utero and with adult female length. Furthermore there were also strong relationships between the peripheral eosinophilia with serum (P<0.01) and gastric mucus IgA activity (P<0.01). Moreover serum pepsinogen and the number of L4 at slaughter were related (P<0.01).

The liver fluke Fasciola hepatica is a worldwide distributed trematode parasite of great veterinary importance. In Spain, the prevalence of the infection is up to 56% in sheep. Its control is mainly based on the use of anthelmitics, although some resistant populations have been described. Genetic studies of the genus Fasciola have been focused on the 28S ribosomal deoxyribonucleic acid (rDNA) gene, with no intraspecific variations and a few interspecific nucleotide differences. We have designed a specific polymerase chain reaction to amplify and sequence a 510-bp-long fragment of the gene to characterize several F. hepatica ovine isolates from the northwest of Spain. All the isolates rendered the expected results on the agarose gel, while sequencing revealed some differences in the 34 site. The anthelmintic-resistant specimens showed the same nucleotide differing of the sensitive ones. Comparing to other studies, our results conclude that there are some genetic variations in F. hepatica (at the level of the 28S rDNA), including heterozygous specimens. In conclusion, a genetic heterogeneity has been found in the F. hepatica populations of ovine origin in the northwest of Spain, using an assay that could be useful for further epidemiology and characterization studies.

The aim of this experiment was to find an antigen from Teladorsagia circumcincta to measure the level of IgA in ovine serum samples. Previous experiments used the titre of IgA to select sheep resistant to infection with T. circumcincta. By Western blotting 22 experimentally infected ewes recognized many proteins, but not all of them were recognized by all animals. Sheep were divided into two groups of animals: those that recognized a protein in Western blotting and those that did not. Among all proteins, three showed significant associations between protein recognition and the titre of IgA in the serum.

SUMMARY The aim of this study was to understand the influence of a low protein diet on the resistance of the Churra breed sheep to infection with Teladorsagia circumcincta. A previous experiment in our department demonstrated significant differences between resistant and susceptible sheep infected with T. circumcincta, in eggs per gram of faeces, serum pepsinogen, serum IgA and peripheral eosinophilia (unpublished). The sheep were fed with a balanced protein diet. In the current assay there were significant differences between the resistant and susceptible groups in egg output, serum pepsinogen and worm burden. The optical densities (OD) of the IgA in blood samples, nasal secretions and saliva were tested throughout the study and the differences between groups were not significant. At necropsy adult worm length, the number of eggs in utero in adult females, the number of fourth stage larvae and the OD of gastric mucus IgA did not differ between groups either. Moreover, due to diet there was a loss of weight in both groups but this was not significant. Our results support the view that low protein diets could influence immune function and as a result resistant genotypes do not show any superiority in comparison with susceptible ones.

The objectives of this study are to find an appropriate ELISA technique to monitor IgA activity during infection by Teladorsagia circumcincta in sheep as well as to identify the degree of resistance to the infection. Previous studies have shown the relationship between IgA in infected sheep and the resistance to the infection. In this study, the titre of IgA in resistant and susceptible infected ewes was tested in blood, nasal secretions and saliva. ELISAs were carried out against somatic antigen from L4 and against a 203 amino acid fragment of a recombinant protein disulphide isomerase of T. circumcincta (Tc-PDI). The results showed that, in general, the titres of IgA against a fragment of a Tc-PDI were lower than against the somatic antigen. These lower levels are clearly shown in blood samples and nasal secretions in contrast to saliva samples where the titre reached higher values testing the IgA against Tc-PDI. This could be due to the fact that, in the detection of IgA against Tc-PDI, the antibody-antigen reaction is more specific than against the whole somatic antigen. To differentiate between resistant and susceptible animals, the significant differences were more present with the somatic antigen. Nevertheless, the ELISA technique using the somatic antigen cannot be standardized, and the antigen-antibody reactions are more unspecific; therefore, the recombinant Tc-PDI could be a better option.