S.A. Bisset

Breeding sheep in New Zealand that are less reliant on anthelmintics to maintain health and productivity

Abstract Breeding sheep that are less reliant on the use of anthelmintics to maintain acceptable health and productivity is one of several options that may assist farmers to manage the growing anthelmintic-resistance problem in New Zealand and meet consumer demands to minimise drug usage in livestock. Although it has been known for many years that genetic factors contribute to the ability of sheep to cope with roundworm challenge, attempts to selectively breed for such factors have occurred only recently. Two host traits, ‘resistance’ and ‘resilience’, are thought to be involved in limiting the deleterious effects of roundworms on the health and productivity of sheep, and the associated need for treatment with anthelmintics. Over the last decade, considerable advances have been made in our understanding of the feasibility and implications of breeding for these traits under practical conditions. The experimental breeding lines developed as part of this research are now proving to be a valuable resource for further work directed at identifying genes and/or genetic markers associated with host resistance and understanding the mechanisms of host immunity to nematode parasites in ruminants. This review provides an up-to-date summary of the results of research in this field, with particular reference to dual-purpose sheep in New Zealand, and describes how this information is being applied in practical breeding programmes.

Managing anthelmintic resistance: Is it feasible in New Zealand to delay the emergence of resistance to a new anthelmintic class?

Abstract The recent registration in New Zealand of the first new class of broad-spectrum anthelmintic, for use against nematode parasites of ruminants, in nearly three decades has raised the possibility that parasite management practices could be improved to minimise the emergence of resistance to the new drug. A review of knowledge pertaining to the selection of anthelmintic resistance in nematode parasites of sheep highlights a number of management practices which could be altered to achieve this. A number of previously common practices such as whole-flock treatment of adult ewes around lambing, and treatment of lambs as they are moved onto pastures with low parasite contamination have been clearly identified as high risk for selecting resistant parasites. Once high-risk practices have been identified steps can be taken to either eliminate their use or mitigate the associated risk. Much of the focus on the management of resistance around the world is on the retention of susceptible genotypes in refugia. While approaches to retaining unselected parasites are likely to vary around the world, empirical studies indicate that the practice is likely to be effective at slowing the development of resistance. The challenge for farmers and advisors will be to strike a balance between retaining sufficient susceptible para-sites to usefully delay the development of resistance while not unduly compromising animal performance and farm profitability. The merits of combining different classes of anthelmintic in order to slow the development of resistance remains somewhat contentious in some countries. However, the attributes of oral anthelmintics are such that they seem likely to meet most, if not all, of the criteria for combinations to be highly effective at slowing the build-up of resistance in nematode parasites. It is evident that considerable progress has been made in understanding the factors involved in selecting anthelmintic-resistant nematodes since the last broad-spectrum anthelmintic class was released in the early 1980s. Therefore, it should be possible to manage a new class of anthelmintic in such a way as to significantly extend its effective life. The challenge is likely to be in convincing farmers of the merits of adopting such pro-active strategies.

Discovery of quantitative trait loci for resistance to parasitic nematode infection in sheep: I. Analysis of outcross pedigrees

BackgroundCurrently most pastoral farmers rely on anthelmintic drenches to control gastrointestinal parasitic nematodes in sheep. Resistance to anthelmintics is rapidly increasing in nematode populations such that on some farms none of the drench families are now completely effective. It is well established that host resistance to nematode infection is a moderately heritable trait. This study was undertaken to identify regions of the genome, quantitative trait loci (QTL) that contain genes affecting resistance to parasitic nematodes.ResultsRams obtained from crossing nematode parasite resistant and susceptible selection lines were used to derive five large half-sib families comprising between 348 and 101 offspring per sire. Total offspring comprised 940 lambs. Extensive measurements for a range of parasite burden and immune function traits in all offspring allowed each lamb in each pedigree to be ranked for relative resistance to nematode parasites.Initially the 22 most resistant and 22 most susceptible progeny from each pedigree were used in a genome scan that used 203 microsatellite markers spread across all sheep autosomes. This study identified 9 chromosomes with regions showing sufficient linkage to warrant the genotyping of all offspring. After genotyping all offspring with markers covering Chromosomes 1, 3, 4, 5, 8, 12, 13, 22 and 23, the telomeric end of chromosome 8 was identified as having a significant QTL for parasite resistance as measured by the number of Trichostrongylus spp. adults in the abomasum and small intestine at the end of the second parasite challenge. Two further QTL for associated immune function traits of total serum IgE and T. colubiformis specific serum IgG, at the end of the second parasite challenge, were identified on chromosome 23.ConclusionDespite parasite resistance being a moderately heritable trait, this large study was able to identify only a single significant QTL associated with it. The QTL concerned adult parasite burdens at the end of the second parasite challenge when the lambs were approximately 6 months old. Our failure to discover more QTL suggests that most of the genes controlling this trait are of relatively small effect. The large number of suggestive QTL discovered (more than one per family per trait than would be expected by chance) also supports this conclusion.

Epidemiology of nematodosis in Romney lambs selectively bred for resistance or susceptibility to nematode infection

Field trials were undertaken to compare nematode population dynamics, lamb productivity and levels of breech soiling in experimental flocks of Romney lambs selectively bred for increased resistance or susceptibility to nematode infection. In each year of the 2 year study, spring-born ewe lambs derived from Wallaceville Animal Research Centres divergent nematode-resistant and nematode-susceptible breeding lines were grazed as separate flocks on matched farmlets from weaning (at 3 months old) until they were approximately 10-11 months old. Allocation of farmlets was reversed between Years 1 and 2 of the study to account for any possible paddock-related effects. Within each year both flocks were subjected to identical management conditions, including anthelmintic treatment (which was administered only when the overall mean faecal worm egg count measured across both genotypes reached 1500 eggs g-1). In both years, by mid-autumn (April) nematode larval infestation levels on pasture were approximately 5-6-fold greater on the farmlet grazed by susceptible (S) genotype lambs than on that grazed by their resistant (R) counterparts (Year 1: 2506 cf. 544 larvae kg-1 herbage; Year 2: 431 cf. 74 larvae kg-1 herbage). This led to 51-fold and 56-fold differences in faecal egg count between R and S lambs by late autumn (May) and winter (July) in Years 1 and 2, respectively. Although mean growth rates were similar in the R and S lambs over summer (while pasture infestation levels on the farmlets were still in the process of diverging), significantly higher growth rates occurred in the R than in the S lambs over autumn-winter in both years of the study (P < 0.01). In contrast, no significant differences in growth rate occurred in either year between male lambs derived from the nematode-resistant and nematode-susceptible breeding lines which were grazed together on another area of the Wallaceville farm from weaning until late autumn. Despite the substantially lower pasture infestation levels encountered by the R ewe lambs, they nevertheless temporarily suffered more breech soiling (dags) than their S counterparts (P < 0.01) in both years. Yearling fleece-weights of the R and S genotypes did not differ significantly in either year. Although the results of our study confirmed that there are potentially significant epidemiological benefits to be derived from breeding sheep for resistance to nematode infection, these benefits did not appear to be associated with large advantages in animal performance. Further work is needed to establish how these results should be interpreted with respect to anthelmintic drench requirements of genetically resistant animals.

Genetic and phenotypic relationships among Trichostrongylus colubriformis-specific immunoglobulin E, anti-Trichostrongylus colubriformis antibody, immunoglobulin G1, faecal egg count and body weight traits in grazing Romney lambs

Variation in total and Trichostrongylus colubriformis-specific immunoglobulin E (IgE) was investigated in four- and six-month old Romney lambs reared on pasture, where they were exposed to natural challenge with nematode parasites. The lambs, which were from an experimental progeny test flock (n=64 sires), were born in spring over three years and weaned at an average age of three months (December). Data from the flock were analysed to obtain heritability estimates for IgE and genetic correlations between IgE and other immunological, parasitological and production traits. In addition, correlated responses of IgE to long-term genetic selection for high, control or low faecal nematode egg count (FEC) were investigated in a set of experimental breeding lines. Repeatabilities and heritabilities of IgE traits in the progeny-test flock were similar to those calculated previously for anti-T. colubriformis antibody (Ab), immunoglobulin G1 (IgG1) and FEC (generally >0.30). Genetic correlations of the loge transformed IgE traits (January and March samples) with loge (FEC+100) on three sampling occasions were all negative (−0.22 to −0.37). However, positive correlations (0.17 and 0.43) were found between loge IgE and dag score, indicating a tendency for there to be more severe breech soiling in lambs with elevated serum IgE. In the 1995 and 1996 selection-line lamb crops (after 17 and 18 years of selective breeding for high or low FEC), both total and T. colubriformis-specific IgE levels were higher (by between 59 and 103%) in the low than in the high line (P<0.001). IgE levels in the controls were intermediate between those of the high and low lines but closer to those of the low line. The results are in line with other evidence which suggests that greater genetic resistance to gastrointestinal nematode infection in Romney lambs is associated with an increase in activity of the TH2 arm of the immune system which mediates inflammatory responses against multicellular parasites.

Resistance to therapeutic treatment with macrocyclic lactone anthelmintics in Ostertagia circumcincta.

Eighty-eight lambs were allocated to one of four groups which were then dosed with 10,000 infective-stage larvae (L3) of one of four populations of Ostertagia circumcincta; the first (S) was an isolate known to be anthelmintic-susceptible; the second (OR) was a multiple anthelmintic-resistant isolate which had been recovered from the field following therapeutic failure of both ivermectin and moxidectin and subsequently maintained in the laboratory without further anthelmintic selection. The third (R) was derived from OR but had been passaged for five generations in the laboratory with each generation being screened with all three broad-spectrum drench families; the fourth (R x S) was an F1 cross between the S and R isolates. On patency, each of the four infection groups was sub-divided into five treatment groups, one of which received no anthelmintic while the others were administered either oral ivermectin (IVM-oral), controlled-release capsules containing ivermectin (IVM-CRCs), oral moxidectin (MOX-oral) or injectable MOX (MOX-inj). Neither formulation of IVM reduced FEC in the R, R x S and OR infected lambs compared to their untreated controls, but significant reductions were observed in all cases following MOX-oral or MOX-inj treatment. Similarly, neither IVM formulation significantly reduced the numbers of R or R x S worms compared to their untreated controls, although the numbers of OR worms were reduced in both cases (P<0.05). Direct comparisons of efficacy across the isolates, however, indicated that neither formulation was any more effective against R x S or OR worms than against the more highly selected R worms. In contrast, both MOX formulations significantly reduced worm numbers of all the resistant isolates compared to their respective untreated controls; furthermore, worm burdens of R x S were reduced significantly more than burdens of R (P<0.05). Reductions in OR burdens, which were intermediate between the two, did not differ significantly from either. The results are consistent with published work on Haemonchus contortus, which suggests that macrocyclic lactone (ML) resistance is expressed as a dominant trait under treatment with IVM. However, these data differ from the H. contortus studies in suggesting that ML resistance in O. circumcincta may effectively be rendered incompletely dominant or recessive by treatment with MOX.

Resistance to prophylactic treatment with macrocyclic lactone anthelmintics in Teladorsagia circumcincta.

Sixty-four Romney ewe lambs were allocated to 12 groups on the basis of liveweight. Four groups (n=5) were administered oral ivermectin (IVM), 4 (n=6) oral moxidectin (MOX) and the remaining 4 (n=5) controlled-release capsules containing IVM (IVM-CRCs). Nine and 10 days later, the groups within each treatment type were challenged with infective-stage larvae (L3) of 1 of 4 different isolates of Teladorsagia circumcincta (two doses each of 5000 L3). The first of these (S) was known to be anthelmintic-susceptible; the second (OR) was a multiple anthelmintic-resistant strain recovered from the field following therapeutic failure of both ivermectin and moxidectin and subsequently maintained in the laboratory without further anthelmintic selection; the third (R) was derived from OR but had been passaged for five generations indoors with each generation being screened with all three broad-spectrum anthelmintic classes; and the fourth (RxS) was an F1 cross between the R and S isolates. As anticipated, because of its limited residual activity, IVM had no significant effect on the establishment, 9 and 10 days post-treatment, of any of the parasite isolates. In contrast MOX, which has greater residual activity, was highly effective at preventing the establishment of the S isolate but showed no significant effect against the OR, R or RxS isolates. The IVM-CRC was also highly effective at preventing the establishment of the S isolate and furthermore it significantly reduced establishment of both the OR and RxS isolates, although it had no significant effect against the R isolate. The results suggest that with respect to the establishment of T. circumcincta L3s following anthelmintic treatment, macrocyclic lactone (ML) resistance is effectively a dominant trait in the presence of MOX, while it behaves as a partially dominant/recessive trait under treatment with IVM-CRCs. The potential implications of this finding in relation to selection for ML resistance in T. circumcincta are discussed.

Selecting for resilience in Romney sheep under nematode parasite challenge, 1994–2007

Abstract Selective breeding for increased resilience to nematode parasite challenge in New Zealand Romney sheep is reported for the period 1994–2007. Resilience was defined as the age at which a first post-weaning anthelmintic treatment was required to maintain acceptable growth in lambs grazing nematode-contaminated pasture. An experimental line selected for resilience (Rsl) was established in 1994. This was expanded in 1997 for a study comparing animal performance under conventional and low-chemical management systems. An elite resilient line (ER) was re-established in 1999. Selection increased the average ‘age-at-first-drench’ in ER lambs by 23.6 days relative to their control line counterparts. This was accompanied by a 4.5 kg increase in 6-month live weight, and a significant reduction in breech-soiling at 5 months of age. No significant change in faecal worm egg count occurred. Although the Rsl lambs still required some anthelmintic intervention under low-chemical management, the study nevertheless provided useful insights into the potential value of resilience to the sheep industry. Some practical difficulties currently associated with on-farm selection of lambs for resilience may be overcome if appropriate selectable genetic markers or biomarkers can be identified.

Anthelmintic resistance to two drench families in a dairy goat herd: Suggestions for future control options

Abstract Extract Madam:— In early March 1988 a dairy goat farm in the Wellington area was visited, at the request of the owner, to investigate the possible involvement of nematode parasitism in causing persistent diarrhoea in several of the does.

Genomic introgression mapping of field-derived multiple-anthelmintic resistance in Teladorsagia circumcincta.

Preventive chemotherapy has long been practiced against nematode parasites of livestock, leading to widespread drug resistance, and is increasingly being adopted for eradication of human parasitic nematodes even though it is similarly likely to lead to drug resistance. Given that the genetic architecture of resistance is poorly understood for any nematode, we have analyzed multidrug resistant Teladorsagia circumcincta, a major parasite of sheep, as a model for analysis of resistance selection. We introgressed a field-derived multiresistant genotype into a partially inbred susceptible genetic background (through repeated backcrossing and drug selection) and performed genome-wide scans in the backcross progeny and drug-selected F2 populations to identify the major genes responsible for the multidrug resistance. We identified variation linking candidate resistance genes to each drug class. Putative mechanisms included target site polymorphism, changes in likely regulatory regions and copy number variation in efflux transporters. This work elucidates the genetic architecture of multiple anthelmintic resistance in a parasitic nematode for the first time and establishes a framework for future studies of anthelmintic resistance in nematode parasites of humans.