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.     

Determination of anthelmintic efficacy against equine cyathostomins and Parascaris equorum in France

This article reports the results of a faecal egg count reduction test on 4 farms in France, as an integrated part of the routine deworming strategy against horse cyathostomins and Parascaris equorum. Treatment with fenbendazole (FBZ) or ivermectin (IVM) was evaluated in yearlings on Farms 1 and 2 and treatment with pyrantel embonate (PYR) was tested on Farms 3 and 4. Calculation of the arithmetic mean faecal egg count reduction and the 95% confidence intervals (95% CI) around the mean was performed using bootstrap analysis. For equine cyathostomins, resistance to FBZ was found with an arithmetic mean reduction of 48.8% (95% CI: 1.9–69.3%). On Farms 1 and 2, horses with reduced efficacy were identified. PYR was found to be effective against cyathostomins, with an arithmetic mean reduction of 95.3% (95% CI: 84.6–99.8%), as well as IVM (100%). For P. equorum, both FBZ and PYR were effective (100% reduction). The efficacy of IVM, however, was low (45.5%; 95% CI: 0–96.3%). These results confirm that FBZ resistance in equine cyathostomins is present in France and that anthelmintic resistance to IVM is present in P. equorum. This study underlines the necessity to evaluate the efficacy of horse deworming strategies on a regular basis under field conditions.     

Managing anthelmintic resistance: Modelling strategic use of a new anthelmintic class to slow the development of resistance to existing classes

AIM: To test the hypothesis that a single strategic treatment with a new class of anthelmintic could slow the development of resistance to existing classes of anthelmintic.

METHODS: An existing model was used to simulate nematode parasite dynamics and the development of anthelmintic resistance. Variations on a five-drench preventive programme of treatments for lambs, in which either zero, the first, third or fifth treatment was substituted with a different class of drug, were compared for the time to reach treatment failure (defined as efficacy <95%). The sensitivity to variations in the death rate of adult worms, that varied from 1 to 5%, and the dominance of resistance genes were also assessed.

RESULTS: Replacing one of the five treatments with a different class of anthelmintic almost always slowed the development of resistance, and was never worse than using the same drug for all treatments. Further, there were large differences in the relative time to treatment failure depending on which treatment was substituted. Changing the first treatment always had the least benefit, whereas changing the fifth treatment always had the greatest. This pattern was independent of the daily death rate of adult worms, and was not influenced by the dominance of resistance under treatment.

CONCLUSIONS: The results indicated that strategic substitution of a single treatment with a new class of anthelmintic, at the end of a series of preventive treatments to lambs using an existing class, could slow the further development of resistance to the latter. This strategic use of a new anthelmintic class has the potential to greatly extend the life of existing anthelmintics if these are still effective.     

Has the optimum time for faecal nematode egg count reduction testing in sheep in New Zealand changed?

Abstract

AIM: To determine what, if any, changes have taken place in the optimum time, for undertaking faecal egg count reduction tests (FECRT) in sheep in New Zealand.

METHODS: A comparison was made between the numbers and types of nematode genera adequately represented for testing purposes (faecal nematode egg count (FEC) of >50 epg) in initial FECRT case submissions to a veterinary laboratory in New Zealand, during two 4-year periods, in 1992–1995 and 2006–2009.

RESULTS: Although there were some minor differences between them, the seasonal patterns of occurrence remained basically the same for all parasite genera in both datasets, with their individual peak periods of representation occurring during February to May in all instances. Not surprisingly, this period of maximum seasonal occurrence for each parasite genus also coincided with those months of the year when the greatest numbers of worm genera were adequately represented for faecal nematode egg count reduction (FECR) testing.

CONCLUSIONS: The results of this study indicate that the optimum time for conducting FECRT in sheep in New Zealand continues to be during the late summer-autumn months of February to May. However, they also reaffirmed that even during this optimal period there are still likely to be many occasions when relatively few nematode genera may be sufficiently well represented for satisfactory FECR testing. Accordingly, veterinary practitioners ought to be aware that, in order to obtain a complete picture of the resistance status of all worm genera on a particular property, such testing may need to be carried out on more than one occasion.     

用丙硫苯咪唑检测羊线虫抗药性的体外虫卵孵化试验

为探讨用丙硫苯咪唑进行体外虫卵孵化试验检测羊线虫对苯并咪唑类药物的抗药性,对23个羊场的25份田间样品用丙硫苯咪唑和噻苯咪唑进行了虫卵孵化试验并与先前减卵试验（faecal egg count re-duction test,FECRT）的结果比较。结果表明,25份样品中,丙硫苯咪唑和噻苯咪唑对受检虫卵的半数致死量（LD50）均值分别为0.050 1和0.054 0μg/mL,差异不显著。FECRT检测有抗药性的4个羊场,75%的样品对丙硫苯咪唑和噻苯咪唑的LD50均值均在0.1μg/mL以上,只有1个羊场的的LD50值分别为0.068 9μg/mL（丙硫苯咪唑）和0.071 2μg/mL（噻苯咪唑）。检测为可疑的2个羊场,其样品的LD50值为0.04～0.07μg/mL;FECRT检测敏感的蠕虫群体的LD50均在0.04μg/mL以下。此外,丙硫苯咪唑用纯的二甲基亚砜溶解和稀释后于4℃保存7 d,LD50值变化不大,提示药效无明显下降,而保存14 d后药效有下降趋势。     

An evaluation of anthelmintic properties,assessed using faecal nematode egg counts,of New Zealand native flax (Phormium tenax)

AIM: To investigate the anthelmintic properties of New Zealand native flax (Phormium tenax) for cattle, using a faecal egg count reduction test (FECRT).

METHODS: Twenty-six heifer calves with high (>300 epg) faecal nematode egg counts (FEC) were paired into two groups and fed either chopped flax leaves ad libitum, or barley straw at an equivalent dry matter (DM) rate, from Day 0–7; all were supplemented with 1.5 kg/head/day cereal feed supplement. On Days 8–10, all heifers were fed a common diet of 3.5 kg DM/ head/day barley straw and 1.5 kg/head/day cereal feed supplement. Heifers were weighed and faecal samples were collected on Days —6, 0, 7, and 10; samples were analysed in triplicate for FEC. The nutritive value of the flax and straw was analysed.

RESULTS: Both the flax and straw had low dry matter digestibility (DMD) and protein content. Although the flax-fed claves grew more than the controls, this may have been related to gut fill. Total daily faecal egg output was similar for flax and straw groups on Day 7 (14.7 vs 15.0 x 10⁶ eggs/day, respectively) and Day 10 (14.9 vs 15.1 x 10⁶ eggs/day, respectively). There was no difference in the change in FEC with time between the calves fed flax or straw diets.

CONCLUSION: Consumption of flax leaves did not reduce FEC in calves with a mixed nematode infection.     

Comparison of in vitro methods and faecal egg count reduction test for the detection of benzimidazole resistance in small strongyles of horses

The objective of the study was to compare the in vitro egg hatch test (EHT), larval development test (LDT) and in vivo faecal egg count reduction test (FECR test) for the detection of benzimidazole resistance in equine strongyles. The presence of resistant or susceptible strongyle populations was determined in 25 stud farms using the in vivo FECR test and in vitro EHT. On the basis of the FECR values, resistance to fenbendazole was detected on 15 of the 25 farms (60%). The ED₅₀ value (anthelmintic concentration producing 50% inhibition of hatching) for suspected resistant populations varied from 0.110 to 0.222 g/ml thiabendazole (TBZ). Final LD₅₀ values (anthelmintic concentration inhibiting development of 50% of eggs into L₃ infective larvae) above 0.029 g/ml TBZ in the in vitro larval development test on samples from 11 stud farms revealed the presence of populations of small strongyles suspected of being benzimidazole-resistant.     

Field studies investigating anthelmintic resistance in young cattle on five farms in New Zealand

AIM: To investigate the efficacy of pour-on anthelmintics against field strains of parasitic nematodes in young cattle on five farms in New Zealand.

METHODS: Faecal nematode egg count (FEC) reduction (FECR) tests were carried out on five calf-rearing farms using pour-on formulations of levamisole, ivermectin, eprinomectin, and the simultaneous administration of levamisole and ivermec- tin. Faecal samples were collected per rectum before treatment and about 7, 14, 21 and 28 days after treatment, for FEC and faecal nematode larval culture.

RESULTS: Resistance (i.e. <95% reduction in FEC) of Cooperia oncophora to ivermectin and eprinomectin was identified on all five farms. There was limited evidence of possible emerging resistance in Ostertagia spp to ivermectin but not eprinomectin, in short-tailed larvae of Cooperia spp to ivermectin and eprinomec- tin, and in Trichostrongylus spp to ivermectin, eprinomectin and levamisole used separately. Levamisole was effective against C. oncophora, but had variable efficacy against Ostertagia spp in the calves in this study. Simultaneous treatment with levamisole and ivermectin pour-on formulations were effective against all genera on all farms.

CONCLUSIONS: To effectively manage roundworm parasites in their calves farmers need to be aware of the resistance status of the parasites on their farms. Levamisole is likely to be an effective anthelmintic on most farms at times of the year when the impact of Ostertagia spp is not high. Simultaneous administration of levamisole and ivermectin pour-on anthelmintics to cattle is likely to control both ML-resistant C. oncophora and stages of Ostertagia spp that are not controlled by levamisole alone.     

Faecal worm egg count analysis for targeting anthelmintic treatment in horses: Points to consider

Equine gastrointestinal nematodes are ubiquitous; in horses that graze contaminated pasture and that are not treated appropriately, large numbers of worms can accumulate, which can lead to serious clinical disease. Nematode control has traditionally followed interval treatment regimens, which involve regular anthelmintic administration to all horses based on the strongyle egg reappearance periods of each drug, usually defined around the time of licensing. Interval treatment programmes have resulted in substantial reductions in large strongyle disease, but have made major contributions to the development of anthelmintic resistance, particularly in cyathostomins. Cyathostomin resistance to 2 of the 3 available anthelmintic classes is widespread, and resistance to both classes in single populations is not uncommon. Reduced efficacy of the most commonly used macrocyclic lactone anthelmintics, as measured by shortened egg reappearance periods after treatment, is emerging in cyathostomins. Macrocyclic lactone resistance is also now commonly reported in Parascaris equorum on stud farms. Faecal worm egg counts (FWEC) are increasingly being used as part of targeted approaches to parasite control, whereby only those horses with moderate to high FWEC within a group are treated with an anthelmintic. The objective of this approach is to reduce environmental contamination, while leaving a proportion of the worm population in some horses unexposed to selection pressure for anthelmintic resistance. This article reviews recent findings in equine parasitology research that will underpin guidelines for control, with a particular focus on how to optimise the value of FWEC methodologies and anthelmintic efficacy analyses.     

A survey on anthelmintic resistance in nematode parasites of sheep in the Slovak Republic

The prevalence of anthelmintic resistance on 27 sheep farms in Slovakia was investigated in 2003 and 2004 using the faecal egg count reduction test (FECRT) according to the WAAVP guidelines. Resistance to albendazole was detected on one farm (3.7%) and suspected on two farms (7.4%) out of 27 sheep flocks. Resistance to ivermectin was tested on 26 farms. On six (23.1%) farms, results indicated the presence of ivermectin resistance. Resistance to ivermectin was suspected on eight farms (30.8%). However, it is also possible that generic ivermectin anthelmintics used in survey have a lower efficacy against sheep nematodes.     

The relationship of training milestones with racing success in a population of Standardbred horses in New Zealand

AIM: To gather information on the repeatability of a faecal nematode egg count (FEC) reduction (FECR) test (FECRT), evaluating both different methods of calculating efficacy and variations within a method, in order to supply veterinarians and other advisors with sufficient information to apply some level of confidence around a diagnosis of anthelmintic resistance based on FECRT results.

METHODS: Two commercial sheep farms were selected on the basis of having previously recorded FECR <95% after treatment with ivermectin (Farm 1) or albendazole (Farm 2). On each farm at least 250 lambs, managed as a single mob, were individually ear-tagged and sampled for FEC. The resulting counts were used, 3—4 days later, to sort the lambs into 24 groups of 10. First, the animals were split into three groups of 80, having high, medium or low FEC. Second, within each of these groups the 80 animals were further divided into four replicate mobs of 20 (each with the same mean count). Third, each of these replicates was further split into two groups of 10: those that would be drenched and those that would remain as untreated controls. All animals were again faecal-sampled and those in the drenched groups were dosed, using a syringe, to their individual liveweight, with ivermectin (Farm 1) or albendazole (Farm 2). Ten days after treatment all animals were individually faecal- sampled again. FEC and larval cultures were undertaken for all 24 groups from both pre- and post-treatment samples. Ef- ficacy (FECR) of the undifferentiated FECRT was calculated using three different equations, and efficacy by genus was also calculated.

RESULTS: Calculated efficacies differed between equations, and the equation which did not utilise an untreated control yielded significantly lower efficacy estimates on both farms. Faecal cultures varied considerably in the proportions of parasite genera recovered. In general, this did not differ between FEC groups, except on Farm 1 where Haemonchus spp were more common and Cooperia spp less common in high-FEC samples. Estimated efficacies against individual genera varied considerably or very little, depending on the level of resistance. On both farms, differing proportions of tests against some genera passed or failed FECRTs based on a threshold pass mark of ≥95% FECR.

CONCLUSION: There was considerable variability in the outcomes of FECRTs and in larval culture results. Caution is warranted in interpreting the results of FECRTs when efficacy values fall into the 90—95% range. Further, the possibility of a test returning a false-negative result is raised, indicating that even an efficacy estimated ≥95% may not guarantee the absence of resistant parasites.     

Minimising the development of anthelmintic resistance, and optimising the use of the novel anthelmintic monepantel, for the sustainable control of nematode parasites in Australian sheep grazing systems

Objective To compare the risk of different treatment scenarios on selecting for anthelmintic resistance on Australian sheep farms. Design A computer simulation model predicted populations of Trichostrongylus colubriformis, Haemonchus contortus or Teladorsagia (Ostertagia) circumcincta, and the frequency of anthelmintic resistance genes. Method Nematode populations and the progression of drug resistance for a variety of treatment options and management practices in sheep‐rearing areas of Western Australia (WA), Victoria (VIC) and New South Wales (NSW) were simulated. A scoring system was devised to measure the success of each option in delaying resistance to each anthelmintic and in controlling nematode populations. Results The best option at all sites was combining the new anthelmintic (monepantel) with a triple mixture of benzimidazole, levamisole and abamectin (COM). The next best option was: in NSW, rotation at each treatment between monepantel, moxidectin and COM; in VIC, rotation at each treatment between monepantel and COM; and in WA, rotation at each treatment between monepantel (used in winter) and COM or moxidectin (used in summer–autumn). In WA, rapid selection for resistance occurred as a consequence of summer–autumn treatments; however, if a small percentage of adult stock were left untreated then this selection could be greatly reduced. Despite purposely assuming relatively high resistance to benzimidazole and levamisole, COM was still effective in controlling worms and delaying resistance. Conclusions Because of cost constraints, it may not be feasible or profitable for producers to always use the combination of all drugs. However, the second‐ and third‐best options still considerably slowed the development of anthelmintic resistance.     

A multi-species model to assess the effect of refugia on worm control and anthelmintic resistance in sheep grazing systems

Objective Develop a computer simulation model that uses daily meteorological data and farm management practices to predict populations of Trichostrongylus colubriformis, Haemonchus contortus and Teladorsagia (Ostertagia) circumcincta and the evolution of anthelmintic resistance within a sheep flock. Use the model to explore if increased refugia, provided by leaving some adult sheep untreated, would delay development of anthelmintic resistance without compromising nematode control. Procedures Compare model predictions with field observations from a breeding flock in Armidale, NSW. Simulate the impact of leaving 1–10% of adult sheep untreated in diverse sheep‐grazing systems. Results Predicted populations of Tr. colubriformis and T. circumcincta were less than those observed in the field, attributed to nutritional stress experienced by the sheep during drought and not accounted for by the model. Observed variation in faecal egg counts explained by the model (R²) for these species was 40–50%. The H. contortus populations and R² were both low. Leaving some sheep untreated worked best in situations where animals were already grazing or were moved onto pastures with low populations of infective larvae. In those cases, anthelmintic resistance was delayed and nematode control was maintained when 1–4% of adult stock remained untreated. Conclusions In general, the model predicted that leaving more than 4% of adults untreated did not sufficiently delay the development of anthelmintic resistance to justify the increased production risk from such a strategy. The choice of a drug rotation strategy had an equal or larger effect on nematode control, and selection for resistance, than leaving 1–10% of adults untreated.     

A Field Study on the Effect of Some Anthelmintics on Cyathostomins of Horses in Sweden

The objective of the study was to investigate different aspects on the efficacy of three anthelmintics on cyathostomin nematodes of Swedish horses. A faecal egg count reduction (FECR) test was performed on 26 farms. Horses were treated orally with recommended doses of ivermectin, pyrantel pamoate or fenbendazole. Faecal samples were collected on the day of deworming and 7, 14 and 21 days later. No resistance was shown against ivermectin; the FECR was constantly >99%. The effect of pyrantel was assessed as equivocal in 6 farms 14 days after treatment; the mean FECR was 99%. As many as 72% of the fenbendazole-treated groups met the criteria for resistance; the mean FECR was 86%, ranging from 56% to 100%. A re-investigation of two farms where pyrantel resistance had been suspected clearly revealed unsatisfactory efficacy of pyrantel on one of these farms; the FECR varied from 72% to 89%. Twenty-six of the horses previously dosed with pyrantel or fenbendazole, and which still excreted ≥150 eggs per gram of faeces 14 days after treatment, were dewormed with ivermectin and fenbendazole or pyrantel in order to eliminate the remaining cyathostomins. A total of 13 cyathostomin species were identified from horses that initially received fenbendazole and seven species were identified from pyrantel-treated individuals. The egg reappearance period (ERP) following treatment with ivermectin and pyrantel was investigated on two farms. The shortest ERP after ivermectin treatment was 8 weeks and after pyrantel was 5 weeks. We conclude that no substantial reversion to benzimidazole susceptibility had taken place, although these drugs have scarcely been used (<5%) in horses for the last 10 years. Pyrantel-resistant populations of cyathostomins are present on Swedish horse farms, but the overall efficacy of pyrantel is still acceptable.     

An evaluation of techniques used for the detection of anthelmintic resistance in nematode parasites of domestic livestock

In vitro and in vivo techniques used for the detection of anthelmintic resistance are reviewed in terms of their versatility, reliability, accuracy, cost and simplicity. The faecal egg count reduction test is considered the most suitable for field screening of resistance but the larval development assay is likely to prove a valuable adjunct in such investigations. Other in vitro techniques, such as egg hatch tests, larval motility and tubulin binding assays are recommended as suitable only for research investigations. Similarly, in vivo procedures such as the critical and controlled anthelmintic efficacy tests are considered to be appropriate only for specific research purposes because of the costs involved in such studies.