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.     

Probable multigeneric resistance to macrocyclic lactone anthelmintics in cattle in New Zealand

AIM: To establish the efficacy of topical formulations of eprinomectin and abamectin against naturally acquired abomasal and small intestinal nematode infections in cattle purchased from a North Island bull-beef property. METHODS: A controlled slaughter trial, involving eighteen 6-8-month-old mixed breed calves, was conducted in May 2002.The animals were randomly allocated on the basis of faecal egg count to one of three equal-sized groups (n=6), consisting of an untreated control group and two treatment groups. One of the treatment groups was treated with a topical formulation of eprinomectin, the other with abamectin. Both anthelmintics were administered as a single topical treatment on an individual liveweight basis, at the manufacturer's recommended dose rates of 0.5 mg/kg. All calves were housed in separate groups with no access to pasture throughout the entire trial and were slaughtered 7-10 days after treatment. RESULTS: Both anthelmintic treatments were highly effective(worm count reduction >98%) against Ostertagia ostertagi, Trichostrongylus axei and Cooperia punctata, but were not effective at reducing worm counts significantly of either Cooperia oncophora or Trichostrongylus longispicularis. Against these latter two parasites, worm count reductions of only 72% and 79%, and 81% and 76%, respectively, were recorded following treatment with eprinomectin or abamectin, respectively (all p>0.05). CONCLUSIONS: These results demonstrate evidence of resistance to macrocyclic lactone anthelmintics by C. oncophora and probably T. longispicularis also. CLINICAL RELEVANCE: As well as perhaps providing the first record of resistance to any anthelmintic by T. longispicularis, the present findings may also represent the first case of resistance to macrocyclic lactone anthelmintics exhibited by more than one parasite species at a time in cattle in New Zealand. KEYWORDS: Eprinomectin, abamectin, macrocyclic lactone, anthelmintic resistance, cattle, Cooperia, Trichostrongylus.     

Resistance to macrocyclic lactone anthelmintics by Haemonchus contortus and Ostertagia circumcincta in sheep in New Zealand

AIM: To establish the efficacy of oral formulations of ivermectin and moxidectin against naturally acquired abomasal nematode infections on a North Island sheep farm. METHODS: Two controlled slaughter trials were undertaken. In the first, 30 sheep on pasture were randomly allocated on the basis of faecal egg count to 1 of 3 groups, comprising an untreated control group and 2 treatment groups. One treatment group was given a single oral dose of ivermectin and the other a single oral dose of moxidectin, both at the manufacturer's recommended dose rates of 0.2 mg/kg liveweight. Six days after treatment, all animals were slaughtered and their abomasa recovered for worm counting. The second trial, which involved 47 animals, was essentially the same as the first except that, as well as involving the slaughter of 30 sheep from all 3 groups, 6 days after treatment, it also included a further 8 untreated control animals and 9 moxidectin treated animals which were slaughtered 27 days after treatment. RESULTS: At 6 days after treatment, moxidectin was highly effective against all 3 of the abomasal nematodes present. While ivermectin was similarly effective against Trichostrongylus axei 6 days after treatment, it was not effective against either Ostertagia circumcinta or Haemonchus contortus, against which average efficacies of only 63.6% and 61.6%, respectively, were recorded. At 27 days after treatment, moxidectin, was also highly effective against T. axei (97.3% reduction) but not against either H. contortus (71.4% reduction) or O. circumcinta (61.0% reduction). CONCLUSIONS: These results provide the first record of macrocyclic lactone resistance in H. contortus in sheep or in any other host in New Zealand, and the first case where such resistance has been exhibited in more than one parasite species at a time. Although the therapeutic efficacy of moxidectin was high against these resistant H. contortus and O. circumcincta strains, resistance to moxidectin was indicated by its diminished prophylactic activity against them. It is suggested that this reduction in the prophylactic activity of moxidectin is also likely to reduce its apparent current high therapeutic efficacy. CLINICAL RELEVANCE: As well as providing further evidence that it can no longer be automatically assumed that macrocylic lactone anthelmintics will be effective on sheep farms in this country, these findings also present a warning that increasingly complex parasite control options may have to be faced in the future.     

Resistance to macrocyclic lactone anthelmintics and associated risk factors on sheep farms in the lower North Island of New Zealand

AIM: To estimate the prevalence of resistance to macrocyclic lactone (ML) anthelmintics over a 3-year period on sheep farms in the Taihape area, and to determine associations between the presence of ML resistance and farm management practices. METHODS: All farmers with >1,000 lambing ewes served by one veterinary practice in the Taihape area of the central North Island of New Zealand were invited to take part in the study (n=157); respondents were selected on their willingness to participate. Resistance to ML was measured during 2004-2006, using a standard faecal egg count reduction test (FECRT) and a half dose of oral ivermectin (IVM; 0.1 mg/kg; n=84). Additional FECRTs were conducted separately on some farms, using full doses of IVM (n=47), levamisole (LEV; n=49), a benzimidazole (BZ; n=76), and a BZ/LEV combination (n=42). Resistance was defined as <95% faecal nematode egg count reduction (FECR) 7-10 days post-treatment. Larval cultures were performed on pooled faecal samples from all pre- and post-treatment groups for which any positive faecal nematode egg counts (FEC) were recorded. To ascertain whether particular management or drenching strategies were associated with the presence of resistance, a questionnaire comprising 50 questions was developed, and each farmer interviewed individually. RESULTS: Of all farmers invited, 84/157 (54%) participated in this study. Based on undifferentiated FEC, ML resistance to the half-dose IVM was evident on 46/84 (55%) participating farms. The majority of these cases (71%) involved Ostertagia (Teladorsagia) circumcincta. Resistance was also evident to full doses of IVM on 22/47 (47%), BZ on 41/76 (54%), LEV on 18/49 (37%), and BZ/LEV on 2/42 (5%) farms. Positive associations (p<0.05) were evident between the presence of ML resistance and management practices, such as: weaning over half of the lambs onto paddocks not grazed by lambing ewes since June; not always returning lambs to the same paddock after drenching; and the use of visual signs to assess 'worminess'. Measuring drench efficacy in January to March rather than later in the year, and presence of resistance to BZ or LEV were associated with a higher apparent prevalence of ML resistance (p<0.05). CONCLUSIONS: Anthelmintic resistance, and particularly ML resistance, was widespread on sheep farms in the Taihape area, and mostly involved O. circumcincta. The prevalence of resistance was lower on farms on which management practices, particularly those involving lambs, maintained refugia of unselected nematodes. Further investigation is required to develop control strategies which minimise selection for resistant worms, and how these may vary with climate.     

Resistance to benzimidazole anthelmintics in field strains of Ostertagia and Nematodirus in sheep

SUMMARY: Resistance to a range of benzimidazole anthelmintics was investigated in 2 strains of Ostertagia spp. One strain (SRBO) had been exposed to fenbendazole, oxfendazole and thiabendazole, the other strain (KR79) only to thiabendazole. Both strains showed a high degree of resistance to albendazole, fenbendazole, oxfendazole and thiabendazole, which had efficiencies of 11–38% and 0–60% against all developmental stages of SR80 and KR79, respectively. There was no significant reduction in the KR79 worm count by thiabendazole at 132 mg kg-1 given either as a single dose or divided into 12 equal portions, one administered every 6h. Nematodirus spp were also found to be resistant to all benzimidazoles tested. Levamisole at 7 mg kg^-1 and naphthalophos at 30 mg kg-1 had efficiences of 89 and 66%, respectively, against SR80 Ostertagia and 99 and 19% against Nematodirus, all but the last of these being significant reductions in worm burdens. The resistant SR80 Ostertagia occurred on a research station, but possibly originated from a property which 3 years earlier had supplied ewes to the station. The use of benzimidazole anthelmintics and subsequent grazing on worm-free pasture may have enhanced the level of resistance. A relationship was established between egg counts and adult worm counts 10 days post-treatment, which suggested that for Ostertagia the worm count could be predicted from the geometric mean egg count from about 10 animals. Thus, where an Ostertagia population is suspected of being resistant, an anthelmintic efficiency assay using pre-and post-treatment faecal egg counts should provide a satisfactory diagnostic procedure.     

Correlation between loss of efficacy of macrocyclic lactone heartworm anthelmintics and P-glycoprotein genotype

Macrocyclic lactone (ML) molecules have been used for heartworm control for more than 25 years. However, in recent years, there have been reports of loss of efficacy of ML heartworm preventatives against Dirofilaria immitis in some locations in the United States. Macrocyclic lactone resistance is a common problem in nematode parasites of livestock, and more recently, evidence of ivermectin resistance has been reported in the human filarial nematode Onchocerca volvulus. In this study, four D. immitis sample groups from the United States with different treatment histories were investigated for evidence of ML-driven genetic selection. DNA from individual adult worms and microfilariae was amplified by polymerase chain reaction to investigate a gene encoding a P-glycoprotein, a protein class known to be involved in ML pharmacology. A significant correlation of a GG-GG genotype with ivermectin response phenotype was found. Moreover, a significant loss of heterozygosity was found in a low responder group; loss of heterozygosity is commonly seen in loci when a population has been under selection. Further studies are required to confirm ML resistance in heartworm populations. However, the genetic changes observed in this study may be useful as a marker to monitor for ML resistance in D. immitis.     

Strategic use of anthelmintics for parasitic nematodes in cattle and sheep

Treatment regimes for the control of parasitic nematodes in cattle and sheep are reviewed. The advantages and disadvantages of each strategy are outlined and the veterinarian's judgement of the management and economic constraints involved when recommending a particular system to a client is emphasised.     

Resistance to benzimidazole anthelmintics by trichostrongyles in sheep and goats in north-west Cameroon.

Following the routine use of tiabendazole at monthly intervals for several years, the trichostrongyle parasites of sheep and goats on a government research station in North-West Cameroon had become resistant to benzimidazole anthelmintics.     

Multigeneric resistance to benzimidazole anthelmintics in sheep

Benzimidazole resistance involving several gastrointestinal nematode genera on a single sheep farm in New Zealand is reported for the first time. A controlled slaughter trial showed that at the recommended dose rate of 12.5 mg/kg, mebendazole had efficacies of 0, 60, 66, 90, 54 and 38% against Haemonchus contortus, Ostertagia spp., Nematodirus spp., intestinal Trichostrongylus spp., Strongyloides spp. and Oesophagostomum venulosum, respectively. The relevance of such multigeneric resistance to possible future options for controlling anthelmintic resistant sheep trichostrongylids is briefly discussed.     

Ivermectin-resistant Ostertagia circumcincta from sheep in the lower North Island and their susceptibility to other macrocyclic lactone anthelmintics

Aim: To confirm the ivermectin resistance status of a strain of Ostertagia circumcincta which was isolated from a sheep farm in the lower North Island of New Zealand and to assess the susceptibility of this strain to other macrocycliclactone anthelmintics.

Methods: Twenty-five lambs housed indoors were each infected with 12,000 L3 larvae of the above parasite strain. Approximately 3 weeks after infection the lambs were allocated to 1 of 4 treatment groups (3 groups of 6, and 1 group of 7 lambs), one of which remained untreated while the others were drenched orally with ivermectin, moxidectin or abamectin at 0.2 mg/kg liveweight. Faecal egg counts (FECs) before and after treatment, and post-mortem worm burdens 10 days after treatment were examined to assess efficacies of each anthelmintic.

Results: Treatment with ivermectin reduced the mean FEC by only 18% and the mean worm burden by only 42%, whereas moxidectin and abamectin reduced FECs by > 92% and worm burdens by >95%.

Conclusion: These results, together with a similar case described recently from the South Island, confirm the emergence of ivermectin resistance in nematode parasites of sheep in New Zealand. The superior efficacy of moxidectin and abamectin in this case indicates that, following the emergence of resistance to ivermectin, some short-term practical use may still be made of these other anthelmintics. However, their continued use will undoubtedly result in increased levels of resistance and eventual therapeutic failure of these products also.     

Ivermectin-resistant Ostertagia circumcincta from sheep in the lower North Island and their susceptibility to other macrocyclic lactone anthelmintics

AIM: To confirm the ivermectin resistance status of a strain of Ostertagia circumcincta which was isolated from a sheep farm in the lower North Island of New Zealand and to assess the susceptibility of this strain to other macrocycliclactone anthelmintics. METHODS: Twenty-five lambs housed indoors were each infected with 12,000 L3 larvae of the above parasite strain. Approximately 3 weeks after infection the lambs were allocated to 1 of 4 treatment groups (3 groups of 6, and 1 group of 7 lambs), one of which remained untreated while the others were drenched orally with ivermectin, moxidectin or abamectin at 0.2 mg/kg liveweight. Faecal egg counts (FECs) before and after treatment, and post-mortem worm burdens 10 days after treatment were examined to assess efficacies of each anthelmintic. RESULTS: Treatment with ivermectin reduced the mean FEC by only 18% and the mean worm burden by only 42%, whereas moxidectin and abamectin reduced FECs by 92% and worm burdens by 95%. CONCLUSION: These results, together with a similar case described recently from the South Island , confirm the emergence of ivermectin resistance in nematode parasites of sheep in New Zealand. The superior efficacy of moxidectin and abamectin in this case indicates that, following the emergence of resistance to ivermectin, some short-term practical use may still be made of these other anthelmintics. However, their continued use will undoubtedly result in increased levels of resistance and eventual therapeutic failure of these products also.     

A survey of the prevalence of emerging macrocyclic lactone resistance and of benzimidazole resistance in sheep nematodes in the lower North Island of New Zealand

AIM: To investigate the occurrence of emerging macrocyclic lactone (ML) resistance and of resistance to benzimidazole anthelmintics on a number of sheep farms in the North Island of New Zealand. METHODS: On commercial sheep farms (n=30) in the Taihape district in the North Island of New Zealand, 30 animals were randomly allocated to one of two equal-sized groups and treated with either half of the recommended dose rate of ivermectin (half of 0.2 mg/kg), or with the full recommended dose rate of oxfendazole (4.5 mg/kg). The ivermectin treatment only was used on a further six properties. Faecal egg counts, accompanied by pooled larval cultures, were conducted on all samples at the time of treatment and 7-10 days later. RESULTS: Resistance, as indicated by a <95% faecal egg count reduction (FECR) in both instances, was found to oxfendazole on 13/30 (43%) farms and to a half dose of ivermectin on 12/36 (33%) properties. For oxfendazole, such resistance was found to involve all six nematode genera whereas for ivermectin it was almost entirely restricted to Ostertagia and Cooperia infections. CONCLUSIONS: These results indicate that emerging ML resistance may be more common on sheep farms in New Zealand than is generally realised. They also suggest that the half-dose ivermectin faecal egg count reduction test (FECRT) may offer some very practical benefits for parasite control by providing early warning of developing resistance to ML drenches and by signalling the possible imminent failure of these at their therapeutic dose rates. The sensitivity and reliability of this procedure may be further enhanced by the inclusion of larval cultures.     

A survey of the prevalence of emerging macrocyclic lactone resistance and of benzimidazole resistance in sheep nematodes in the lower North Island of New Zealand

AIM: To investigate the occurrence of emerging macrocyclic lactone (ML) resistance and of resistance to benzimidazole anthelmintics on a number of sheep farms in the North Island of New Zealand.

METHODS: On commercial sheep farms (n=30) in the Taihape district in the North Island of New Zealand, 30 animals were randomly allocated to one of two equal-sized groups and treated with either half of the recommended dose rate of ivermectin (half of 0.2 mg/kg), or with the full recommended dose rate of oxfendazole (4.5 mg/kg). The ivermectin treatment only was used on a further six properties. Faecal egg counts, accompanied by pooled larval cultures, were conducted on all samples at the time of treatment and 7–10 days later.

RESULTS: Resistance, as indicated by a <95% faecal egg count reduction (FECR) in both instances, was found to oxfendazole on 13/30 (43%) farms and to a half dose of ivermectin on 12/36 (33%) properties. For oxfendazole, such resistance was found to involve all six nematode genera whereas for ivermectin it was almost entirely restricted to Ostertagia and Cooperia infections.

CONCLUSIONS: These results indicate that emerging ML resistance may be more common on sheep farms in New Zealand than is generally realised. They also suggest that the half-dose ivermectin faecal egg count reduction test (FECRT) may offer some very practical benefits for parasite control by providing early warning of developing resistance to ML drenches and by signalling the possible imminent failure of these at their therapeutic dose rates. The sensitivity and reliability of this procedure may be further enhanced by the inclusion of larval cultures.     

Resistance to benzimidazole anthelmintics in small strongyles (Cyathostominae) of horses in Denmark.

This study was undertaken to establish whether anthelmintic resistance was present in nematode parasites of horses in Denmark. Sixteen horse farms were selected for faecal egg count reduction (FECR) tests to measure the efficacy of the anthelmintic used. Resistance to benzimidazole anthelmintics was found on 13 of the 16 farms, with FECR values ranging from 80.0% to -101.3%. On the remaining 3 farms FECR was 100.0%, 99.3% and 97.2%. Results of a questionnaire study on anthelmintic usage, parasite control measures and management practices showed that horses in this study were treated on average 7.1 times/year. Horse owners changed between preparations of drugs but almost only within the same class of anthelmintics. Nine owners gave an anthelmintic treatment to purchased horses before they were introduced on the farm. On 14 farms, the same paddock was grazed every year and the average stocking rate was estimated to be 2.4 horses/ha. Strategies to avoid development of anthelmintic resistance are discussed and recommendations of parasite control on horse farms are presented.     

Multigeneric resistance to benzimidazole anthelmintics in four sheep flocks

Clinical parasitism associated with multigeneric resistance to benzimidazole anthelmintics was investigated in four sheep flocks during February and March 1989. The genera considered resistant were Haemonchus and Nematodirus spp. on all four farms and Trichostrongylus and Ostertagia spp. on three of the four farms. It is suggested that when investigating cases of suspected drench resistance, postdrenching worm counts are useful to help identify the resistant worm species present.     

Preliminary trials using a macrocyclic lactone against psoroptic scabies of cattle

A macrocyclic lactone, LL-F28249 alpha, C36H52O8, was evaluated as a one- or two-injection treatment in different formulations and concentrations against cattle infested with Psoroptes ovis (Hering). One injection of 2 mg kg-1 or two injections of 1 mg kg-1 of 3% active material in either an aqueous or oil formulation failed to eliminate all of the mites. A single injection of 3 mg kg-1 as a 5% aqueous formulation eliminated all mites for 35 days post-injection. At lower dosages of the aqueous formulation (3%), the male mites appeared to be less susceptible to the acaricide than either the female or immature mites.