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.     

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.     

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.     

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。     

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.     

Molecular mechanisms for anthelmintic resistance in strongyle nematode parasites of veterinary importance

Veterinarians rely on a relatively limited spectrum of anthelmintic agents to control nematode parasites in domestic animals. Unfortunately, anthelmintic resistance has been an emerging problem in veterinary medicine. In particular, resistance has emerged among the strongyles, a group of gastrointestinal nematodes that infect a variety of hosts that range from large herbivores to small companion animals. Over the last several decades, a great deal of research effort has been directed toward developing an understanding of the mechanisms conferring resistance against the three major groups of anthelmintics: macrocyclic lactones, benzimidazoles, and nicotinic agonists. Our understanding of anthelmintic resistance has been largely formed by determining the mechanism of action for each drug class and then evaluating drug‐resistant nematode isolates for mutations or differences in expression of target genes. More recently, drug efflux pumps have been recognized for their potential contribution to anthelmintic resistance. In this mini‐review, we summarize the evidence for mechanisms of resistance in strongyle nematodes.     

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.     

Assessment of anthelmintic resistance in nematode parasites of sheep and goats owned by smallholder farmers in eastern Ethiopia

The anthelmintic resistance status was investigated of nematode parasites of sheep and goats owned by smallholder farmers in communities that received breeding stock from a source where a high level of anthelmintic resistance has been reported. The investigation used the faecal egg count reduction technique, whereby suitable animals within each of eight separate communities were pooled to achieve the numbers required to conduct separate tests for both sheep and goats. Anthelmintics tested were albendazole (ABZ), tetramisole (TET), a combination (ABZ $+$ TET) and ivermectin (IVM), at the manufacturers' recommended dose rates. Results showed that there was no evidence of anthelmintic resistance in nematode parasites of either sheep an goats in any community. This indicates that dilution of resistant parasites imported with introduced breeding stock, and the low selection pressure imposed by the smallholder farmers themselves, has prevented anthelmintic resistance from emerging in nematode parasites of small ruminants in these communities.     

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-levami- sole 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 impres- sion 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.     

Anthelmintic resistance in New Zealand

Anthelmintic resistance was first confirmed in New Zealand in 1979 and since then has become common-place; more than 50 % of sheep farms now have detectable levels of resistance to one or more chemical classes of anthelmintic. Farmer drenching practices have changed little over the last 15–20 years and are clearly exerting a significant level of selection for resistance. In the absence of new chemical classes of anthelmintics, current parasite control practices will be unsustainable in the long-term. Once substantial resistance has developed, significant reversion to susceptibility is unlikely and re-introduction of failed drugs is likely to result in the rapid re-emergence of control problems. The number of anthelmintic treatments applied is not necessarily a reliable indicator of selection pressure and should not be the only factor considered in strategies for minimising the development of resistance. The relative potential of the different anthelmintics now available, particularly the long- acting products, to select for resistance varies with the way they are used and with other epidemiological and management factors; generalisations about their respective roles in the development of resistance are often unreliable. In many cases, literal extrapolation of recommendations for the management of resistance from Australia to New Zealand is unsupportable, given the differences in climate, parasite ecology and farming practices between the 2 countries. In the absence of a refuge for susceptible genotypes, as occurs when anthelmintic treatments are used as a means of generating low-contamination ‘safe’ pasture for young stock, the rapid development of resistance is likely. Anthelmintic treatments applied to animals with a high level of immunity, or which become immune while the anthelmintic is active, are likely to select for resistance faster than treatments applied to non-immune stock.     

A survey on anthelmintic resistance in nematode parasites of sheep in Lusaka,Zambia

While surveys in Southern Africa indicate anthelmintic resistance of gastrointestinal nematodes to be common in small ruminants in South Africa, Kenya and Zimbabwe there have been no reports of resistance in Zambia. The objective of this study was to determine whether anthelmintic resistance occurs in Zambia, and to obtain information on nematode control practices in the country. During the rainy season six commercial sheep farms were selected in and around Lusaka and Chisamba. Worm control practices were gauged by means of a questionnaire, and the Faecal Egg Count Reduction Test was performed for assessing anthelmintic efficacy of albendazole, levamisole and ivermectin. On all farms anthelmintic treatment was the only approach used to control nematode infections. Frequency of treatment varied from twice a year to every 6 weeks and drugs of different anthelmintic groups were alternated within the same year. There was a wide range in faecal egg counts of individual sheep before treatment, with some individual counts of up to 87000. Larval identification showed that Haemonchus was almost the only genus recovered from the faecal cultures before and after treatment. Albendazole resistance was found on five of the six farms. On each of the four farms where ivermectin gave less than 95% reduction in egg counts, there was resistance to albendazole as well. Levamisole showed an efficacy of 95% or higher on all six farms.     

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 macrocy- clic lactone (ML) anthelmintics (n=32). Resistance to an an- thelmintic 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 ≥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 iver- mectin, 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 nema-todes 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.     

Use of two in vitro methods for the detection of anthelmintic resistant nematode parasites on Slovak sheep farms

A survey of the prevalence of anthelmintic resistant nematode populations was conducted on 32 sheep farms in the Slovak Republic. In vitro egg hatch test and larval development tests were used for the detection of resistance to benzimidazole anthelmintics and they were compared with in vivo faecal egg count reduction tests. There was agreement in the declaration of resistance between the faecal egg count reduction test and both in vitro tests. The presence of resistant populations was determined on two farms using egg hatch test. In both farms, the LD(50) values were higher than 0.1 microg TBZ/ml, indicating resistance. By using LD(99) values it might be possible to reveal relatively small proportion of resistant larvae in the population. The prevalence of benzimidazole resistance has not change on Slovak sheep farms during last decade.     

An egg-hatch assay for resistance to levamisole in trichostrongyloid nematode parasites

An in vitro technique is described for detecting resistance of nematodes to the anthelmintic levamisole hydrochloride. Samples of eggs are developed under controlled temperature conditions until just prior to the commencement of hatching. They are then exposed to different concentrations of the drug and, when hatching is almost complete, the test samples are killed and preserved. The proportion of unhatched eggs at each drug concentration can then be counted at leisure. Provided a suitable range of drug concentrations is chosen for each test isolate, this assay provides results which may be satisfactorily fitted to a log-concentration-probit regression model. Comparisons with in vivo anthelmintic assays have shown that the technique provides an accurate reflection of the resistance status of parasite populations.     

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 ≥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 suffi cient 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.     

A field study on the anthelmintic resistance of Parascaris spp. in Arab foals in the Riyadh region,Saudi Arabia

Background: In the last decade, Parascaris spp. resistance to anthelmintics has been recorded in many countries. In Saudi Arabia, there are limited data available on Parascaris spp. resistance to anthelmintics.

Objective: To determine the current status of ivermectin, abamectin and praziquantel combined, and fenbendazole resistance to Parascaris spp. in horses in Saudi Arabia.

Methods: Three hundred and forty-one foals from eleven different farms were examined by faecal egg count (FEC). The foals were all Arab horses aged 17.2 ± 4.5 (SD) months. Ivermectin (n = 46 foals), abamectin and praziquantel combined (n = 46), and fenbendazole (n = 46) were administered on day 0 and faeces were collected on day 14. The study comprised 41 untreated foals as controls. Animals that have FEC of ≥100 eggs per gram (EPG) were used to measure anthelmintic efficacy. Parascaris spp. populations were considered susceptible when faecal egg count reduction (FECR) was ≥95% associated with a lower 95% confidence limit (LCL) >90%, suspected resistant when FECR ≤90% or LCL <90% and resistant when FECR <90% and LCL <90%.

Results: Prevalence of Parascaris spp. infection was 53% (179/341 horses). Anthelmintic resistance to Parascaris spp. were highest following fenbendazole (55% of farms and 65% of foals) and to a lower extent following ivermectin or the combination of abamectin and praziquantel which comprised 27% of farms (and 46% of foals) and 18% of farms (and 10% of foals), respectively.

Conclusion: These data indicate that anthelmintics-resistant Parascaris spp. populations are present on horse farms in Saudi Arabia.