Haemonchus contortus resistance to ivermectin and netobimin in Brazilian sheep.

Suffolk, Texel, Hampshire Down and Ile de France sheep from the municipalities of Porto Amazonas, Piraquara and Araucaria in the State of Paraná, and Bagé in the State of Rio Grande do Sul were brought to Sobral, State of Ceará, to be used in a cross-breeding project. On arrival they had clinical signs of nematode parasitosis, and one Suffolk female died. The animals were treated orally with ivermectin (0.2 mg kg-1) and fifteen days later with netobimin (20.0 mg kg-1). Neither drug reduced the egg counts (measured in eggs per gram, EPG) significantly, and this suggested that the nematodes in the sheep were resistant to the anthelmintics used. Haemonchus contortus was the species involved. The egg counts were reduced after oral treatment with trichlorfon (100.0 mg kg-1). Haemonchus contortus larvae obtained from these animals before trichlorfon treatment and passaged through two nematode-free sheep were used in a further experiment. Twenty 6- to 9-month-old nematode-free lambs were infected with the H. contortus larvae (10,000 per animal) and after the infection was confirmed, were randomly divided into four groups of five animals. Group I was orally treated with ivermectin at 0.2 mg kg-1, Group II with oral netobimin at 20.0 mg kg-1, Group III with oral trichlorfon at 100.0 mg kg-1 and Group IV was a non-treated control. Egg counts and faecal cultures were taken before dosing on the day of treatment and seven days later when all animals were necropsied and the nematodes were collected from the abomasa and counted.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypergastrinaemia of sheep infected with Haemonchus contortus

Radioimmunoassay of plasma gastrin has shown that a hypergastrinaemia occurs in yearling sheep, after infection with 10(6) infective third stage larvae of Haemonchus contortus. Plasma gastrin concentrations began to rise two to four days after infection, that is, at about the same time as, or slightly before, the abomasal pH was observed to increase.     

Selection for resistance to macrocyclic lactones by Haemonchus contortus in sheep.

An anthelmintic-sensitive Haemonchus contortus strain was selected for moxidectin and ivermectin resistance concurrently for 22 generations. Treatment with 0.002 mg moxidectin/kg BW or 0.02 mg ivermectin/kg BW produced >99% efficacy against the susceptible parent strain passaged for 22 generations without any anthelmintic exposure. However, to obtain similar efficacy the moxidectin-selected and the ivermectin-selected strains of H. contortus required 0.05 mg moxidectin/kg BW or 0.4 mg ivermectin/kg BW. These results indicate that development of resistance to one macrocyclic lactone, simultaneously results in resistance to another macrocyclic lactone. However, rates of resistance development differ between compounds and occurs more slowly with moxidectin than with ivermectin.     

Towards the eradication of Haemonchus contortus from sheep flocks in Sweden

Previous studies have shown that between-year transmission of Haemonchus contortus in Swedish sheep flocks is almost entirely as over-wintered populations within housed animals, and not on pasture. This suggests that eradication of this parasite is a realistic possibility. Thus, two sheep farms with a history of H. contortus infection on the Swedish island of Oland were selected for study. During the winter housing period of 2003/2004 all ruminants (sheep and cattle) on both farms were treated with ivermectin. Monitoring by faecal egg counts and infective larval differentials of ewes and lambs for the subsequent two grazing seasons, together with total abomasal worm counts of 10 lambs from each farm at the end of the first grazing year, showed that this objective was achieved.     

Experimental Haemonchus contortus infection in three breeds of sheep

Nine- to 10-month-old Florida Native, St. Croix and Dorset/Rambouillet sheep were infected with Haemonchus contortus. The primary infection was terminated by anthelmintic treatment 9 weeks later and animals were then reinfected with H. contortus. Dorset/Rambouillet sheep showed higher fecal egg counts and decreased packed cell volumes and plasma protein levels compared with the other two breeds during the primary infection. However, no breed differences were found in total worm burdens in either primary or secondary infection. Differences between breeds were also not detected in lymphocyte responsiveness to parasite antigen, H. contortus specific mucosal antibody levels, numbers of abomasal mast cells and globule leukocytes or abomasal histamine levels.     

Wool production of sheep chronically infected with Haemonchus contortus

A chronic infection was established in a group of twenty 18-month-old non-reproductive Merino ewes by oral administration of 3000 infective H. contortus larvae twice weekly for 12 weeks. Their live-weights and wool production were compared with those of 20 uninfected ewes grazing the same pasture. In the infected sheep, faecal egg counts increased over a period of 9 weeks to reach a mean of 5000 eggs per gram, accompanied by small, but significant effects on packed-cell volume and live-weight. The effects of infection on wool growth were also small, and not statistically significant, although there was evidence of a faster seasonal decline in wool growth in infected sheep. It was concluded that mortality induced by acute infection is the most important economic effect of this parasite.     

Serum and abomasal antibody response of sheep to infections with Haemonchus contortus

Serum and abomasal IgA, IgG and IgM antibody response against adult worm, L3 and egg antigens of Haemonchus contortus was monitored by the ELISA technique after one or two infections with this nematode. Following the first infection, antibody levels in serum did not change materially. After administration of a challenge dose of infective larvae, antibodies of the three immunoglobulin classes in infected animals rose slightly, but this rise appeared later than the fall in the faecal egg counts. In contrast, in abomasal mucosa, IgA anti-larval antibody levels, which did not increase materially after the primary infection, rose rapidly after a transient inhibition when sheep were challenged. A close temporal relationship was observed between the rise in local anti-worm IgA antibodies and the self-cure reaction, but antibody levels fell rapidly after worm diminution. The local antibody response was thus considered to be related to immunity of sheep to H. contortus.     

Functional investigation of a QTL affecting resistance to Haemonchus contortus in sheep

This study reports a functional characterization of a limited segment (QTL) of sheep chromosome 12 associated with resistance to the abomasal nematode Haemonchus contortus. The first objective was to validate the identified QTL through the comparison of genetically susceptible (N) and resistant (R) sheep produced from Martinik × Romane back-cross sheep. The R and N genotype groups were then experimentally infected with 10 000 H. contortus larvae and measured for FEC (every three days from 18 to 30 days post-challenge), haematocrit, worm burden and fertility. Significant differences in FEC and haematocrit drop were found between R and N sheep. In addition, the female worms recovered from R sheep were less fecund. The second step of the characterization was to investigate functional mechanisms associated with the QTL, thanks to a gene expression analysis performed on the abomasal mucosa and the abomasal lymph node. The gene expression level of a candidate gene lying within the QTL region (PAPP-A2) was measured. In addition, putative interactions between the chromosome segment under study and the top ten differentially expressed genes between resistant MBB and susceptible RMN sheep highlighted in a previous microarray experiment were investigated. We found an induction of Th-2 related cytokine genes expression in the abomasal mucosa of R sheep. Down-regulation of the PAPP-A2 gene expression was observed between naïve and challenged sheep although no differential expression was recorded between challenged R and N sheep. The genotyping of this limited region should contribute to the ability to predict the intrinsic resistance level of sheep.     

A novel approach to the control of anthelmintic-resistant Haemonchus contortus in sheep

This paper reports attempts to control a resistant strain of Haemonchus contortus on pasture by replacing it with a susceptible strain. By making use of artificially infected donor sheep, six camps (paddocks) were seeded with a resistant field strain of H. contortus until it was confirmed by means of worm-free tracer lambs that the grazing had become infective. Thereafter, using donor sheep infected with a susceptible laboratory strain of H. contortus for seeding the pasture, attempts were made at various times of the year to replace the resistant strain on the pasture with the susceptible strain in five of the camps. The sixth remained as a control camp, in which no attempt was made to replace the resistant strain. In two of the five test camps, the susceptible strain was introduced in the autumn after 8-10 weeks of nil grazing; in the remaining three camps the introduction was made in spring (two camps) or summer without having a period of nil grazing. The susceptibility of the worm strains introduced initially, as well as of those that developed in the various camps, was gauged both by controlled non-parametric anthelmintic slaughter trials at the beginning and at the conclusion of the trial, and by an in vitro egg hatch test. A reversion to susceptibility occurred in three of the five camps. These included both of the camps infested with the susceptible strain in the spring and one of the two infested in the autumn. Possible reasons for the failure to replace the resistant strain in the remaining two test camps are discussed.     

Haemonchus contortus with low inhibited development in sheep from the highveld of Zimbabwe

The abomasa of sheep grazing on natural pastures in the highveld of Zimbabwe were examined for Haemonchus contortus. Of 304 abomasa, 213 (70%) harboured H. contortus. The worm burden increased during the rains to reach the peak in February-March. This was followed by a decline with low worm loads throughout the dry season. The fourth stage larvae (L4) accounted for 1-7% of the total H. contortus population, except during July and August when they comprised 22 and 54% of the worm burden, respectively. It appears that the inhibition of H. contortus is not common in the commercial farming sector where sheep are treated very frequently.     

Serial passage of Haemonchus contortus in resistant and susceptible sheep

A recently isolated field strain of Haemonchus contortus was passaged through resistant (repeatedly reinfected) and susceptible (immunosuppressed) sheep for six and nine generations, respectively. Infectivity of the resulting serially passaged strains was not significantly different when tested in groups of 1-year-old susceptible sheep.     

Immunoprotective effect of cysteine proteinase fractions from two Haemonchus contortus strains adapted to sheep and goats

A preliminary analysis of the significance of genetic diversity in cysteine proteinase genes has been performed simultaneously in sheep and goats, with regard to the immunological control using these enzymes against haemonchosis. For this purpose, we have studied the cross-immunoprotective effect of cysteine protease-enriched protein fractions (CPFs) in adult worms of two Haemonchus contortus strains from North America and Spain that are adapted to sheep and goats, respectively. Previous genetic analysis of cysteine proteinase genes in both strains has shown that some of loci are polymorphic and these differences are translated into changes in the amino acid sequences. However, our results show that CPFs from H. contortus adult worms have a protective effect against the parasite in both sheep and goats. These results are similar regardless of whether they were obtained from sheep or goat-adapted H. contortus strains, which could be very important in case H. contortus CPFs were commercially used in different countries, as vaccines to prevent the negative effects of this parasite. Interestingly, this experimental inoculation of both species with a heterologous strain of H. contortus contributes to the idea shown in previous studies about how difficult is the interpretation and the comparison of vaccination where strains not adapted to a specific host are used. Therefore, the challenger of using heterologous strains could provide similar results to those observed in immunised animals. This study suggests the possibility of exploring the mechanisms involved in natural protection against non-adapted strains, in order to develop strategies to control haemonchosis.     

Genetic parameters for resistance to Haemonchus contortus in Merino sheep in South Africa

Resistance to natural infections of Haemonchus contortus (wireworm) was studied in Merino sheep. From February 1995 to July 1996 data were collected from the OTK Merino Stud on the farm Wildebeesfontein near Hendrina in Mpumalanga province. From March 1998 to January 2000 another Merino flock at Irene in Gauteng province was included in the study. In both cases, 50 randomly-chosen animals from each group of lambs weaned were sampled weekly for faecal egg count (FEC) until a mean of 2500 eggs per gram faeces (epg) was reached. At this stage, all lambs in the group were sampled for FEC, packed cell volume (PCV) and live mass in the case of the Wildebeesfontein flock. Animals were monitored from weaning to about 1 year of age. Data from 5 different groups of lambs (583 animals and 1722 records in total) with genetic links were accumulated and analysed. Variance components and resulting heritabilities for logFEC and PCV were estimated using a bivariate animal model with repeated records. The estimated heritabilities were 0.24 +/- 0.02 for logFEC and 0.14 +/- 0.02 for PCV. A strong negative correlation (-0.84 +/- 0.06) existed between the 2 traits. The genetic correlation between PCV and live mass for the Wildebeesfontein flock was low (0.28 +/- 0.09), while that between logFEC and live mass was negative, although not significantly different from zero (-0.13 +/- 0.09). These results suggest that FEC can be used as a selection criterion for resistance to H. contortus infections in Merino sheep.