Cross infection of Haemonchus contortus between calves and lambs.

Haemonchus contortus can frequently be found infecting pre-weaned beef calves on sheep and beef farms around the North Island of New Zealand. The purpose of this study was to determine whether parasites cycling in young cattle constitute a potentially important source of infection for sheep. A field isolate of H. contortus was cycled through either calves or lambs for 3 generations. The larvae resulting from the third cycle of infection were then used to infect both lambs and calves and the resulting faecal nematode egg count (FEC), worm burden, adult worm length and in utero egg count were measured. Larvae derived from lambs inoculated into calves exhibited lower establishment rates, the adult worms were shorter, had lower in utero egg counts, and the resulting faecal egg counts were also lower than when inoculated into lambs (p < 0.01). H. contortus' lack of ability to passage freely between lambs and calves indicates that large populations are unlikely to occur under mixed grazing, resulting in limited potential as a source of infection in sheep. However, indications of an ability to adapt to the alternative host suggest that some investigation of infection in cattle dominant farming operations in the north of the country might be warranted.

The production costs of Haemonchus contortus and other nematode parasites in pre-weaned beef calves in New Zealand.

Haemonchus contortus can frequently be found infecting pre-weaned beef calves on sheep and beef farms around the North Island of New Zealand. The purpose of this study was to consider whether the presence of this parasite alone, or as part of a mixed infection, could be impacting growth rates of young animals, on three commercial farms in the North Island of New Zealand. Trials were conducted on commercial sheep and beef farms in each of the Northland, King Country and Gisborne regions, in late summer/autumn (February to April) of 2016 to measure the effect of treatment with narrow and broad spectrum anthelmintics on liveweight gain of spring-born calves pre-weaning. Each farm was chosen based on the presence of Haemonchus and that it was a beef cow/calf system with the cows and calves grazing the same pastures as sheep at some stage. Three sampling visits were made to each farm with the animals being weighed, faecal sampled and treated with one of two anthelmintics (Closantel alone to remove only Haemonchus or a triple combination containing moxidectin, levamisole and oxfendazole to remove all nematodes) or left untreated, on each of the first two visits. There was no significant difference in liveweight gain between any of the treatment groups, hence there was no evidence for an impact of Haemonchus alone, or a mixed nematode infection, on pre-weaned calf growth rates on these farms. It remains unclear whether there may be a justification to consider treatment of calves should they constitute a significant source of pasture larval infestation with H. contortus, in an integrated cattle-sheep system.

Route of administration influences the concentration of ivermectin reaching nematode parasites in the gastrointestinal tract of cattle.

An animal trial was conducted to measure the concentrations of ivermectin occurring in abomasal and small intestinal contents and mucosa, and in the target parasites (Ostertagia ostertagi and Cooperia oncophora) following administration by subcutaneous, oral and pour-on routes. Twenty-five steers were infected with ivermectin-resistant isolates of O. ostertagi and C. oncophora and following patency randomly allocated to 3 treatment groups of 7 and 1 untreated control group of four. On day 0, animals in the treatment groups were administered ivermectin via the oral, injectable or pour-on routes. On days 1, 2, 3, 4, 5, 6 and 8, blood samples were collected from all live animals, one animal from each treatment group was euthanised and the abomasum and small intestine recovered. Control animals were euthanised on each of days 4, 5, 6 and 8. Samples of gastrointestinal tract organs, their contents, mucosa and parasites were collected and assayed for ivermectin concentration using HPLC. The highest plasma concentrations occurred following subcutaneous administration. In the gastrointestinal contents the highest levels occurred following oral administration, although one high value occurred following pour-on administration, which was attributed to self-licking by the treated animal. The lowest GI content levels followed subcutaneous injection. Ivermectin concentrations in the gastrointestinal mucosa were highest following subcutaneous injection. Drug levels in the abomasal parasite O. ostertagi were most closely correlated with levels in the abomasal mucosa whereas levels in the intestinal C. oncophora were most closely correlated with those in the intestinal contents. Thus, the maximun levels of drug reached C. oncophora in the small intestine following oral administration. In contrast, the highest levels of ivermectin in O. ostertagi followed subcutaneous injection. Therefore, route of administration is likely to influence the exposure to ivermectin for different parasite species.

Nematode parasites in young cattle: what role for unexpected species?

AIMS To investigate the timing of infection of beef calves with sheep nematode species on three sheep and beef farms, and to determine the prevalence of cross-infection in calves before weaning across a larger number of farms. METHODS Farms in the Far North, Gisborne and Tararua districts, in the North Island of New Zealand, were enrolled in 2014. Fresh faecal samples were collected from approximately 10 calves on each farm between birth and up to 5 months after weaning. In 2016, faecal samples were collected from calves before weaning from 22 farms across the upper North Island. For both trials faecal samples were assessed for faecal nematode egg counts and cultured to determine parasite genus. For samples from the three farms, larvae were identified to species using a multiplex PCR assay. RESULTS On the three farms, the median percentage of sheep nematode species detected in faecal cultures was 25 (min 3, max 77)%. The main sheep species detected were Cooperia curticei and Haemonchus spp. (putatively contortus). In faecal samples collected before weaning from 22 farms, Haemonchus spp. were present in 19/22 samples, and the median prevalence was 15 (min 0, max 73)% of the total larvae cultured. CONCLUSIONS AND CLINICAL RELEVANCE The implications of sheep nematode species being present in calves should be considered by farmers and veterinarians when undertaking anthelmintic efficacy testing, as they may contribute to false conclusions regarding anthelmintic efficacy. Pre-weaning calves may also be a possible source of contamination and/or refugia for Haemonchus spp. on farms and should be considered when developing parasite control plans for sheep.

The efficacy and plasma profiles of abamectin plus levamisole combination anthelmintics administered as oral and pour-on formulations to cattle.

In phase I, faecal egg count reduction tests (FECRT) were conducted on six commercial cattle farms to compare the performance of two pour-on and one oral combination anthelmintic. Groups of 12-15 calves were sampled for faecal nematode egg count (FEC) before treatment with either abamectin oral, levamisole oral, an abamectin+levamisole oral combination or one of two abamectin+levamisole combination pour-ons. Samples were collected again 14days after treatment to calculate the percentage reduction in FEC. The proportions of infective stage larvae (L3) in faecal cultures were used to apportion egg counts to, and calculate efficacy against, the main parasite genera. Abamectin oral was effective against Ostertagia except on one farm where resistance was indicated, but had reduced efficacy against Cooperia on four farms. Levamisole oral was effective against Cooperia on all farms, but had variable efficacy against Ostertagia. The abamectin+levamisole oral was effective against both species on all farms. The abamectin+levamisole pour-ons were effective on some farms but not on others. In particular, pour-on 2 failed to achieve 95% efficacy in 45% of evaluations, 4/6 against Cooperia and 1/5 against Ostertagia. On some farms the combination pour-ons were less effective than their constituent actives administered alone as orals. In phase II, 8 groups of 6 calves, grazing parasite-free pasture, were infected with putatively ML-resistant isolates of Cooperia oncophora and Ostertagia ostertagi. Once infections were patent groups were treated with oral or pour-on formulations of abamectin alone, levamisole alone, abamectin+levamisole (two pour-ons) or remained untreated. Blood samples were collected for analysis and after 8days all calves were euthanized and abomasa and intestines recovered for worm counts. All treatments were effective against O. ostertagi and all treatments containing levamisole were effective against C. oncophora. Animals treated with the oral combination had higher Cmax and AUC values for abamectin in plasma than animals treated orally with abamectin alone. In contrast, animals treated with the combination pour-ons tended to have lower plasma levels for abamectin than those treated with abamectin alone as a pour-on, with differences in the Cmax and AUC values approaching statistical significance (p-values ≤0.07). There were no differences detected in plasma concentrations of levamisole. The inconsistent and sometimes poor efficacy of the combination pour-ons on-farm is likely due to reduced levels of abamectin in the plasma and hence less active reaching the target worms in the gut.

The in vitro response of field strains of sheep blowflies Lucilia sericata and L. cuprina (Calliphoridae) in New Zealand to dicyclanil and triflumuron.

AIM: To evaluate the in vitro efficacy of triflumuron and dicyclanil against field strains of Lucilia sericata and L. cuprina in New Zealand to establish whether anecdotal reports of declining protection periods afforded by dips containing these active ingredients had validity. METHODS: Strains of field-derived blowflies (Lucilia cuprina and L. sericata) were obtained from adult flies from flytraps or larvae from sheep with flystrike submitted from seven regions of the North and two regions of the South Islands of New Zealand. A total of 27 strains of L. sericata and 19 of L. cuprina were established in culture and first instar larvae obtained were exposed in vitro to dilution series of technical-grade dicyclanil (a pyrimidine carbonitrile) and triflumuron (a benzoyl urea). Concentrations lethal to 50% (LC50) and 95% (LC95) of each strain were calculated and tested by probit analysis, with the ratio of LC50 of each strain divided by LC50 of the most susceptible strain (resistance factor) derived in order to facilitate between-strain comparisons of susceptibility. RESULTS: There was considerable variation between strains of L. sericata larvae in their susceptibility to triflumuron, with those least susceptible originating from the Waikato region. Strains from both Tararua and Manawatu regions were most susceptible and East Coast strains intermediate. All strains of the species tested showed a narrow band of susceptibility to dicyclanil with no regional trends apparent. CONCLUSIONS: Representative field-strains of sheep blowflies exhibited a range of susceptibilities to both triflumuron and dicyclanil, suggestive of both genetic heterogeneity among fly populations and the spectrum of dip use in their source areas. This means there should be concerns for the long-term benefits of benzoyl urea insecticides for sheep farmers in New Zealand, but not yet for dicyclanil-based formulations. CLINICAL RELEVANCE: Flystrike is a debilitating and occasionally lethal disease in sheep such that efficacious sheep dips have prophylactic value in reducing the prevalence of the condition.

Managing anthelmintic resistance--use of a combination anthelmintic and leaving some lambs untreated to slow the development of resistance to ivermectin.

A field study was conducted to test the hypotheses that use of a combination anthelmintic and/or increasing the pool of unselected worms 'in refugia' by leaving a proportion of lambs untreated would slow the development of resistance to ivermectin. Twelve suites of four paddocks (farmlets) were seeded with a mixture of resistant and susceptible isolates of both Trichostrongylus colubriformis and Teladorsagia (Ostertagia) circumcincta calculated to yield a 95% reduction in faecal nematode egg count (FEC) after treatment with ivermectin. Each year for three years the farmlets were stocked in the spring with mobs of lambs which were treated five times at 28-day intervals with either ivermectin or an ivermectin+levamisole combination. In addition, in half the mobs the heaviest 10% of lambs remained untreated at each treatment occasion, resulting in a 2 × 2 factorial treatment structure (i.e. two drench types × two percentage treated) with three complete replicates. The development of resistance to ivermectin, and at the end to levamisole, was measured by larval development assays (LDA) and worm counts from treated and untreated tracer lambs. For T. colubriformis the development of resistance to ivermectin, as measured by tracer lamb worm burdens, was delayed by treatment with the combination and by leaving 10% of lambs untreated. In addition, the interaction between these factors approached significance (p=0.052). Similarly, results of the LDAs indicated a slower development of resistance when lambs were treated with the combination and when 10% of lambs were left untreated. For T. circumcincta, results were compromised by the rapid development of resistance, which appears to be the result of low viability in the field of the susceptible isolate used to contaminate the pastures. Although a small delay in the development of resistance to ivermectin was indicated, this was off-set by an increase in the level of resistance to levamisole. A post-study modelling experiment simulating the conditions of the field study and the starting efficacies for the two nematode species produced equivalent outputs to those measured in the field. Overall, results support the conclusions that use of combination anthelmintics and deliberately increasing 'refugia' of unselected genotypes will slow the development of anthelmintic resistance. However, as indicated in modelling studies, once resistance to all the constituent actives is well developed (efficacy<70%) the value of combinations for slowing the development of resistance is largely lost.

The production cost of anthelmintic resistance in lambs.

The economic impact of anthelmintic resistance was investigated in lambs by comparing productivity parameters in groups of animals treated either with a highly effective anthelmintic, or an anthelmintic to which three species of resistant worms were known to be present. Ten farmlets, each stocked with 30 lambs, were rotationally grazed for 5 months, with monthly treatments of either albendazole, to which resistance existed, or a new combination product containing derquantel and abamectin (DQL-ABA) to which there was no resistance. Stock on five farmlets were treated with each anthelmintic and productivity measures, including liveweights, body condition and faecal soiling were assessed throughout. In addition, fleece weights and information on carcass weight and quality was collected at the end of the trial. Anthelmintic efficacy was measured at the last two treatment dates by faecal egg count reduction test with larval cultures. Albendazole demonstrated efficacies of 48.4% and 40.9% for Trichostrongylus spp. and Teladorsagia circumcincta respectively. By contrast, the DQL-ABA treatments were >99% effective against all genera. The difference in live-weight gain was 9 kg in favour of the DQL-ABA treatments. This translated into a 4.7 kg increase in carcass weight with a 10.4% increase in carcass value. Significant differences in body condition scores, faecal breech soiling and fleece weights were also recorded, all in favour of the DQL-ABA treatments. The time required for 50% of the animals to reach a target live-weight of 38 kg was significantly shorter (by 17 days) in those animals treated with DQL-ABA. The results show that the production cost of subclinical parasitism as a result of using an anthelmintic product which is less than fully effective due to resistance can greatly exceed the cost of routine testing of anthelmintic efficacy and the adoption of new anthelmintic classes. There is a strong case for many farmers to re-evaluate their position on some of these issues in order to optimise financial performance.

Development and spatial distribution of the free-living stages of Teladorsagia circumcincta and Trichostrongylus colubriformis on pasture: a pilot study.

AIMS: To measure the development of Teladorsagia (=Ostertagia) circumcincta and Trichostrongylus colubriformis eggs to third-stage infective larvae (L3) at different times of the year. Also, to measure the spatial distribution of L3 across herbage, soil and faeces, in order to assess whether spatial issues could be important in larval dynamics on pasture. METHODS: Field plots were contaminated with sheep faeces containing approximately 20,000 eggs of each of T. circumcincta and T. colubriformis on five separate occasions, viz 01 December 1996 (summer), 18 March 1998 (autumn), 17 June 1998 (winter), 15 October 1998 (spring), and 23 July 1999 (winter). Replicate plots (n=10) were harvested at intervals for up to 12 months after deposition of faeces, and the number and distribution of L3 were measured. Larvae were sampled from faeces (where these remained), herbage, and three soil zones to a depth of 145 mm. RESULTS: There were large differences between contamination dates in the percentage of eggs that developed to L3. For both species the highest percentage development was for eggs deposited in December (7.8% and 25.9% for T. circumcincta and T. colubriformis, respectively) and the lowest for June (0.4% and 0.03% T. circumcincta and T. colubriformis, respectively). Development in winter was often delayed, and this was always associated with a low yield of larvae, probably due to compounding mortalities associated with long periods of exposure to low temperatures. The relative distribution of L3 present on herbage, in faeces or in the soil varied between sampling times. However, overall the most L3 were recovered from soil (74% and 66% for T.circumcincta and T. colubriformis, respectively, averaged over all samples), and the lowest recoveries were from the herbage. CONCLUSIONS: Although the data are limited, the results indicated that the highest percentage of eggs developed to infective larvae in summer and only minimal development occurred in winter. The data do not support the view that substantial contamination of pastures with sheep parasites occurs over winter. Large numbers of larvae were recovered from soil, which indicates that, assuming they can subsequently migrate onto herbage, soil is a potentially important reservoir of infective larvae in New Zealand. Therefore, the spatial distribution of L3 on pasture may affect both the dynamics and transmission of parasite populations. Further work on both these issues is warranted.

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 1. Patterns of seasonal development.

AIM: To describe the seasonal pattern of development of third-stage infective larvae (L3) from eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus on pasture in the North Island of New Zealand. METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28-31 days after deposition on 13-18 occasions, between January 2005 and July 2006. Analysis of the number of larvae recovered used a mixed model including number of eggs deposited, weight of faeces recovered (an assumed indicator of earthworm activity), site, contamination date, and position of deposited faeces, i.e. on the surface or buried. RESULTS: There was a significant effect of contamination date on development of all three species, with maximum numbers of L3 developing between late spring (November) and early autumn (March), and minimum numbers in June and July. There were large differences between species, with H. contortus exhibiting a long period (April to October) where development was close to zero, whereas T. circumcincta developed to some extent all year round. Development of T. colubriformis was intermediate between the other two species. Burying faeces containing nematode eggs increased the number of L3 recovered compared with surface deposition (p ≤ 0.001), although there were a small number of exceptions involving only T. colubriformis. The weight of faeces recovered at harvest, which was assumed to be an indication of earthworm activity, was correlated with the number of L3 recovered for all species (p<0.001). In a separate analysis, earthworms were assumed to have been active if <5 g faeces remained at harvest. Where this occurred, the number of L3 of T. colubriformis and T.circumcincta recovered was reduced by 56% and 58%, respectively (p<0.001). CONCLUSIONS: A marked seasonal pattern of development was observed for all three species, with the most larvae developing in spring-early autumn and the least in winter. This seasonal pattern was most pronounced in H. contortus and least obvious in T. circumcincta. Burying faeces containing eggs generally resulted in more L3 being recovered, whilst the apparent activity of earthworms resulted in fewer larvae being recovered.

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 2. Weather variables associated with development.

AIM: To identify weather variables associated with the development of eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus to third-stage infective larvae (L3) under a range of climatic conditions on pasture in the North Island of New Zealand. METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28-31 days after deposition, on 13-18 occasions between January 2005 until July 2006. Weather data were recorded at each site, and the association between weather variables and number of L3 recovered was analysed using subsets regression to select best-fitting models from several candidate variables, after adjustment for efficiency of recovery of L3. A multiple linear regression model was then developed for each species, to select weather variables that had both significant and substantive effects on the number of L3 recovered. RESULTS: For all species, mean daily temperature was the best predictor of the number of L3 recovered (p=0.001). For T.circumcincta the final model included mean daily temperature and soil temperature (R²=51%), and for T. colubriformis the model only included mean daily temperature (R²=55%). For development of H. contortus, mean daily temperature was the most significant variable, but moisture in the form of rainfall entropy, i.e. the temporal distribution of rainfall, over the first 14 days was also significant in the final model (R²=34%). CONCLUSIONS: Temperature was the most important determinant of developmental success of free-living nematodes on pasture at the study sites, and probably also for other parts of New Zealand with similar climates. Moisture was not significant in the development of T. circumcincta or T.colubriformis, implying that under the generally moist temperate climate in New Zealand, moisture is seldom limiting for development of these species. Haemonchus contortus appeared to be sensitive to moisture availability and precipitation in the first 14 days after deposition of faeces. The results of this study will be further developed as part of species-specific climate-driven models of parasite development in New Zealand.

Acquired immunity to endoparasites in sheep interacts with anthelmintic treatment to influence selection for anthelmintic resistance.

AIM: To test the hypothesis that anthelmintic treatment of animals with a well-developed immunity to endoparasites will be more selective for anthelmintic resistance than treatment of animals that are not immunocompetent. METHODS: In Experiment 1, five pregnant, mixed-aged ewes, and five newly weaned 3-month-old lambs were housed indoors, and infected three times a week with a mixture of albendazole resistant and -susceptible Teladorsagia (=Ostertagia) circumcincta larvae for 12 weeks. In Experiment 2, six two-tooth ewes, six 9-month-old lambs, and 10 newly weaned 3-month-old lambs were infected three times a week with a mixture of albendazole resistant and -susceptible Trichostrongylus colubriformis larvae for 10 weeks over the winter months of June to August. Egg-hatch assays (EHA) were performed on a weekly basis, to determine the resistance status of eggs passed before and after a treatment with albendazole, that was given at least one week after the infections became patent. RESULTS: Faecal nematode egg counts (FEC) indicated that mature ewes and immunocompetent lambs restricted the establishment of parasite larvae, especially after the anthelmintic treatment. Although EHA were completed for T. circumcincta, the numbers of eggs recovered from the ewes were too few to allow accurate comparisons of anthelmintic resistance status between ewes and lambs. In Experiment 2, viable infections of T. colubriformis only developed in five 9-month-old and five 3-month-old lambs after anthelmintic treatment. Comparison of the concentrations of anthelmintic needed to kill 50% of the eggs (LC(50)) from the lambs showed that where host immunity restricted the establishment of new infection after anthelmintic treatment, the resistance status of eggs passed remained high until the end of the study. In contrast, where new infections established, the resistance status of eggs in the faeces declined to levels close to those prior to anthelmintic treatment. CONCLUSIONS: The results support the conclusion that older animals are more refractory to the establishment of ingested parasite larvae, and that this reduces the dilution of resistant worms surviving an anthelmintic treatment. Surviving resistant worms are therefore likely to make a greater proportional contribution to the resulting population of parasites on pasture. The findings support the view that treating mature sheep with an anthelmintic should be considered a higher-risk practice for selecting anthelmintic-resistant parasites, than treating lambs.

Drench-and-shift is a high-risk practice in the absence of refugia.

AIM: To examine the effect of an anthelmintic treatment to lambs, followed immediately by a shift onto pastures with differing levels of larval contamination, on the development of anthelmintic resistance, in order to support recommendations to farmers regarding drench-and-shift practices for sustainable worm control. METHODS: Newly weaned Romney lambs (n=72) were dosed with third-stage infective larvae (L3) of two nematode parasite species, Teladorsagia (=Ostertagia) circumcincta and Trichostrongylus colubriformis, comprising benzimidazole-resistant and -susceptible isolates, calculated to yield, after treatment with albendazole, a 95% reduction in faecal nematode egg count (FEC). Once infections became patent (Day 0), lambs were randomised into nine groups of eight animals, treated with albendazole at the manufacturer's recommended dose rate, and moved to individual pastures each previously prepared to have one of three different levels of parasite larval infestation (Treatment 1 = low contamination, Treatment 2 = medium contamination, and Treatment 3 = high contamination), and grazed on those pastures before receiving a second treatment with albendazole at Day 47. Anthelmintic resistance status in each group of lambs was measured using FEC reduction (FECR) and egg-hatch assays (EHA) after the first anthelmintic treatment, and FECR after the second treatment. RESULTS: Egg-hatch assays demonstrated significant differences between treatments. The concentration of anthelmintic required to kill 50% of the eggs (LC50) for Treatment 1, comprising the least contaminated pastures, was significantly higher than for Treatments 2 and 3 on Days 33 and 40. Treatment 1 also had a significantly lower FECR at the final anthelmintic treatment, and significantly lower FEC than the other two treatments from Days 26 to 47. CONCLUSIONS: The results demonstrated that the populations of T. circumcincta and T. colubriformis in lambs treated with anthelmintic had significantly higher levels of albendazole resistance at the end of the grazing period in lambs moved onto pastures with relatively low levels of parasite contamination than those moved onto pastures with relatively higher contamination, confirming drench-and-shift onto 'clean' pasture as a high-risk practice for the selection for anthelmintic resistance. While this does not necessarily preclude the use of this practice it does emphasise the importance of taking appropriate remedial action to minimise the risk.

Carbendazim, at concentrations used on pasture for facial eczema control, reduces development of Trichostrongylus colubriformis when sprayed onto infected sheep faeces.

AIM: To determine whether the fungicide, carbendazim, as applied to pastures for controlling facial eczema (FE), would inhibit development of the free-living stages of the gastrointestinal nematode parasite Trichostrongylus colubriformis. METHODS: Two studies were conducted, using sheep faeces containing eggs of T. colubriformis. In the first, the faeces were either exposed or not to an application of carbendazim sprayed at the recommended rate for FE control. After spraying, dishes containing the faeces were incubated at 20 degrees C for 14 days, and the resulting third-stage infective larvae (L3) extracted by baermannisation and counted. In addition, naturally infested pasture was also sprayed, and the number of L3 present 7 days later was assessed by cutting herbage samples and extracting larvae by soaking in water and baermannisation. In the second, the faeces were incubated at 20 degrees C for 0, 3 or 7 days before being exposed to no, one or two applications of carbendazim. After further incubation for 14, 11 or 7 days, L3 were similarly extracted by baermannisation and counted. RESULTS: In the first study, there was a 74% reduction in the number of T. colubriformis larvae recovered from faeces exposed to carbendazim compared with faeces not exposed, but there was no reduction in the number of L3 recovered from herbage. In the second study, faeces incubated for 0 or 3 days prior to exposure to a single application of carbendazim yielded 98% or 89% fewer larvae, respectively, than faeces not exposed. Faeces incubated for 7 days prior to exposure yielded similar numbers of larvae to faeces not exposed. CONCLUSION: Treatment of pastures with carbendazim for FE control is likely to result in reduced development of the larvae of T. colubriformis, and by inference those of other species, where the application coincides with the presence of freshly deposited faeces containing eggs and developing larvae. However, no effect of treatment on L3 was indicated. The significance of this for on-farm nematode parasite control remains to be determined, as does any potential for strategic applications of carbendazim to pasture aimed at reducing numbers of parasite larvae on pasture. The latter should not be contemplated without due consideration of the implications for the development of anthelmintic resistance.

Brave or gullible: testing the concept that leaving susceptible parasites in refugia will slow the development of anthelmintic resistance.

AIM: To test the theory that creating a reservoir of unselected worms by leaving a proportion of lambs in a flock untreated with anthelmintic, i.e. in refugia, will slow the development of anthelmintic resistance in nematode parasite populations. METHODS: Newly weaned Romney lambs (n=180) were infected with two nematode parasite species, Teladorsagia (= Ostertagia) circumcincta and Trichostrongylus colubriformis. For each species, the challenge doses contained a mixture of infective larvae from benzimidazole-resistant and -susceptible isolates calculated to yield, from the combined population, a 95% reduction in faecal nematode egg counts (FEC) following treatment with albendazole. Once the infections were patent, the lambs were divided into nine groups of 20 animals, and each group was allocated to one of three treatments. In Treatments 1, 2 and 3, 100%, 90% and 80% of animals were treated with an anthelmintic, respectively. For treatments 2 and 3, the heaviest animals remained untreated. Following treatment, each group was moved to its own previously prepared low-contamination pasture. Lambs grazed this pasture for 7 weeks before again being treated and moved to new low-contamination pastures (Shift 1 and Shift 2). The parasite populations on pasture resulting from the different treatments were subsequently sampled using tracer lambs, and worm eggs derived from these were used in both egg-hatch assays (EHA) and larval development assays (LDA), to measure albendazole-resistance status. RESULTS: Treating all animals each time the groups were moved to new low-contamination pastures resulted in higher levels of albendazole resistance (p<0.05), measured using EHA and LDA, in subsequent parasite generations than when either 10 or 20% of animals were left untreated. However, higher FEC in the tracer lambs grazed on pastures in Treatments 2 and 3, compared with Treatment 1, indicated an increased level of pasture contamination as a result of leaving some animals untreated. CONCLUSIONS: The results demonstrate that creating a reservoir of unselected parasites slows the development of anthelmintic resistance, and emphasises the risk of treating all animals prior to a shift on to low-contamination pasture. However, higher levels of pasture contamination, resulting from untreated animals, indicate the difficulty in managing both worm control and resistance.

Managing anthelmintic resistance: untreated adult ewes as a source of unselected parasites, and their role in reducing parasite populations.

AIMS: To test the hypotheses that when untreated adult ewes are rotationally grazed (follow behind) on pastures after lambs receiving routine anthelmintic treatments, the ewes can function as a source of unselected parasites in refugia, capable of slowing the development of anthelmintic resistance, and suppress the build-up of parasites resulting from the development of anthelmintic resistance. METHODS: Firstly, the potential of untreated adult ewes to slow the development of anthelmintic resistance, and to suppress parasite populations under differing levels of anthelmintic efficacy, was investigated using a simulation model. Secondly, a field trial with three replicates of each treatment compared two grazing systems (lambs only vs lambs followed by ewes) and two types of anthelmintic, viz albendazole (ALB), to which resistance was present (faecal nematode egg count reduction (FECR)=57-59%) and ivermectin plus levamisole (IL), to which resistance was absent (FECR=97-99%), in a factorial treatment structure. Parasite populations were monitored using faecal nematode egg counts (FEC), faecal larval cultures, pasture larval sampling, and slaughter of tracer lambs. Animal performance was measured using liveweight, dag score, body condition score, and fleece weights. RESULTS: Model simulations indicated that parasites cycling in the untreated ewes could slow the development of resistance being selected for by the anthelmintic treatments given to lambs and this could occur without a nett increase in larval numbers on pasture. Further, as worm control in the lambs declined with increasing levels of anthelmintic resistance the ewes increasingly functioned as nett removers of parasite larvae, effectively reducing parasite population size. In the field trial, untreated adult ewes contributed to pasture infestations of most parasite species, but not Nematodirus spp. Parasite species on pasture and infecting lambs changed when ewes were present, but larval populations on pasture in the autumn were no greater than when lambs grazed alone. In the presence of anthelmintic resistance, parasite populations were reduced when ewes grazed in rotation with lambs, implicating the ewes as nett removers of parasite challenge. CONCLUSIONS: Untreated adult ewes were a source of unselected genotypes, capable of slowing the development of anthelmintic resistance in most, but not all, parasite species. Further, the potential of adult ewes to remove from pasture more parasite larvae than they contribute through faecal contamination indicates a potentially useful role in suppressing parasite populations, particularly when worm control in lambs is less effective as a result of anthelmintic resistance.

Management of gastrointestinal nematode parasites on sheep farms in New Zealand.

AIM: To report current farmer opinions and farming practices relating to control of gastrointestinal nematodes and anthelmintic resistance on sheep farms in New Zealand. METHODS: An interview-based cross-sectional study of grazing management and anthelmintic usage was conducted by veterinarians on 80 randomly selected sheep farms in New Zealand. RESULTS: Useable data were returned by 74/80 (92%) farmers who participated in the study. However, despite contacting 400 farmers the target sample size of 100 farms was not reached. The results indicated that only 31% of farms had previously tested for drench resistance, that effective quarantine-drenching of imported stock was not always carried out, and that farmers were more likely to integrate cattle than ewes into their grazing management of lambs. Furthermore, the number of drenches given to lambs had changed little in 25 years. The use of faecal egg counting by farmers has increased. CONCLUSIONS: Dependence on anthelmintics continues to be high on sheep farms in New Zealand. Whilst the number of drench treatments has changed little, there is more widespread use of persistent or long-acting treatments. Farmers need to be encouraged to monitor the resistance status of nematode populations on their farms and use this information to develop strategies aimed at maintaining susceptible alleles within the parasite populations and conserving the efficacy of existing drug families.

Prevalence of anthelmintic resistance on sheep farms in New Zealand.

AIM: To establish the prevalence of anthelmintic resistance in parasitic nematodes on sheep farms in New Zealand. METHODS: A cross-sectional prevalence study was conducted, using a standardised faecal nematode egg count (FEC) reduction (FECR) test (FECRT) for ivermectin, at a full (0.2 mg/kg) and half (0.1 mg/kg) dose rate, and albendazole, levamisole and albendazole-levamisole in combination, on 60 lambs (n=10 per group) on farms selected from throughout New Zealand. Farms that conformed with selection criteria were chosen at random (n=80) or with a history of suspected resistance to macrocyclic lactone (ML) anthelmintics (n=32). Resistance to an anthelmintic was inferred when there was <95% reduction in FEC 7-10 days after treatment. Larval cultures were performed for all control groups and for treated groups for which resistance was evident. RESULTS: Of the farms randomly selected, 36% showed > or =95% FECR for all anthelmintics tested; resistance to ivermectin at 0.1 and 0.2 mg/kg liveweight was evident on 36% and 25% of these farms, respectively. Resistance to both ivermectin (0.2 mg/kg) and levamisole was evident on 8/80 (10%) farms, to ivermectin and albendazole on 10/80 (13%) farms, and to ivermectin, levamisole and albendazole on 6/80 (8%) farms. The prevalence of resistance to a half dose of ivermectin tended to be more prevalent on farms with a history of suspected ML resistance (p=0.06). Resistance to albendazole was seen across all the main parasite genera, and to levamisole in Nematodirus, Ostertagia (= Teladorsagia) and Trichostrongylus species. Resistance to ivermectin was dominated by Ostertagia spp, although Cooperia, Nematodirus and Trichostrongylus species were also implicated. CONCLUSION: Anthelmintic resistance in parasitic nematodes of sheep is common in New Zealand. Not only was resistance to albendazole and levamisole common, but resistance to the ML, ivermectin, was at a higher prevalence than expected. Sheep farmers and advisors in New Zealand need to re-evaluate the way they manage parasites, and more research is urgently needed if the steady decline in anthelmintic susceptibility is to be halted.

Prevalence of anthelmintic resistance on 62 beef cattle farms in the North Island of New Zealand.

AIM: To establish the prevalence of anthelmintic resistance in parasitic nematodes on a random sample of beef cattle herds in the North Island of New Zealand. METHODS: A cross-sectional prevalence study was conducted using a standardised faecal nematode egg count (FEC) reduction (FECR) test (FECRT) for ivermectin, levamisole and albendazole on 60 calves on each of 62 farms in the North Island chosen at random from farms that conformed with the selection criteria. Resistance to an anthelmintic was inferred when there was <95% reduction in FEC 7-10 days after treatment. Larval cultures were performed for all control groups and for treated groups for which resistance was evident. RESULTS: Of the farms that completed the FECRT, 4/61 (7%) showed > or =95% reduction in FEC for all anthelmintics tested. Resistance to ivermectin was evident on 56/61 (92%) farms, to albendazole on 47/62 (76%) farms, and to both ivermectin and albendazole on 45/61 (74%) farms. Resistance to levamisole was evident on only 4/62 (6%) farms. The parasites most prevalent in resistant populations cultured were Cooperia spp. On 45/61 (74%) farms where Cooperia spp were present in sufficient numbers, resistance to both ivermectin and albendazole was evident. No cases of levamisole-resistant Cooperia spp were detected. Resistance of Ostertagia spp to ivermectin was evident on 4/45 (9%) farms, to albendazole on 15/46 (35%) farms, and to levamisole on 4/46 (9%) farms. CONCLUSION: Anthelmintic resistance in parasitic nematodes of cattle is common in the North Island of New Zealand. Beef farmers need to be aware of the risks posed by anthelmintic resistance, and routine FECR testing is recommended to ensure optimal productivity and to guide decision-making when purchasing anthelmintics to be used on-farm.

Anthelmintic resistance and management of nematode parasites on beef cattle-rearing farms in the North Island of New Zealand.

AIM: To provide information on current farmers' opinions and farming practices thought to be related to anthelmintic resistance, and to test for associations between the presence of anthelmintic resistance and management practices on beef-cattle rearing farms in the North Island of New Zealand. METHODS: A study using an interview-based questionnaire about management of internal parasites was conducted on 62 beef cattle-rearing farms in the North Island of New Zealand, using case-control analyses to test for associations between management practices and the presence or absence of resistance to ivermectin or albendazole. Resistance was inferred from faecal nematode egg count (FEC) reduction (FECR) tests (FECRTs) when there was <90% reduction in FEC 7-10 days after treatment of calves <12 months of age. RESULTS: Of the 59 farmers who completed the questionnaire, most (n=40) ranked parasites highly, and at about the same level as quality and quantity of feed, as important production-limiting factors for their enterprises. In contrast, anthelmintic resistance was not perceived to be a problem on 13 farms, and its importance was rated low on 24, moderate on 15, and high on only six farms. Despite all farms having planned parasite control programmes, there was heavy reliance on clinical signs of parasitism to determine frequency of treatments. About one in three farmers with beef breeder cows routinely treated their calves at marking, one in five treated mixed-age cows, and almost half treated rising 2-year-old cows before calving. One in four farmers used anthelmintics on calves on 8-12 occasions in their first year of life. Co-grazing with other species was rare, but follow-on grazing within 3 months after older cattle or sheep was common. On most farms, grazing cattle was restricted to part of the farm, a finding with implications for parasite control and persistence of larvae in refugia. Macrocyclic lactone (ML) anthelmintics or their combinations with other action families were currently, and for the past 5 years, used more frequently than benzimidazoles and levamisole, and benzimidazole-levamisole combinations. The prevalence of resistance to ivermectin was high (82%) and no plausible model of associations could be constructed from the data. The prevalence of resistance to albendazole was 60%, and the risk of resistance increased as the number of rising 1-year-old cattle present mid-winter increased, and decreased as the number of breeding cows >2 years old present mid-winter increased. CONCLUSION: It is clear that in practice anthelmintic resistance is a secondary consideration to obtaining productivity advantages from the use of anthelmintics in beef cattle. Farmers' opinions were divided on many issues and the overall impression was of confused and diverse thinking regarding the principles of the use of anthelmintics. The overall outlook regarding anthelmintic resistance in cattle is bleak unless the need for integrated and long-term research activities is acted upon soon.