Pyrantel resistance in two herds of donkey in the UK

Resistance to currently available anthelmintics is a serious phenomenon which is prevalent globally. Cyathostomins are one of the major parasites, and are of primary concern in donkeys. There have been reports of emerging resistance to pyrantel, but the status of pyrantel resistance in donkey populations in the UK is largely unknown. This report investigates pyrantel resistance in two geographically isolated donkey herds in the South West of England. The first herd had suspected pyrantel resistance, with already established resistance to other anthelmintics. In the second herd the efficacy of pyrantel was not suspected at the time the study took place. Faecal Egg Count Reduction Test (FECRT) was carried out, revealing large scale resistance. Eighty one percent of the first herd and 73% of the second herd had a FEC of less than 95% after treatment, and anthelmintic resistance was confirmed using the World Association for the Advancement of Veterinary Parasitology guidelines. These findings indicate that anthelmintic resistance to pyrantel exists in both tested donkey populations and illustrate the continuing development of resistance through different classes of chemotherapeutics.     

High level of treatment failure with commonly used anthelmintics on Irish sheep farms

Background

In 2013 a Technology Adoption Program for sheep farmers was established to encourage the implementation of best management practices on sheep farms in Ireland. There were 4,500 participants in this programme in 2013. As part of this programme, farmers had the option to carry out a drench test to establish the efficacy of their anthelmintic treatment.

Results

Flock faecal samples were collected before and after treatment administration and gastrointestinal nematode eggs enumerated. In total there were 1,893 participants in the task, however only 1,585 included both a pre- and post-treatment faecal sample. Of those, 1,308 provided information on the anthelmintic product that they used with 46%, 23% and 28% using a benzimidazole (BZ), levamisole (LEV) and macrocyclic lactone (ML) product respectively. The remaining farmers used a product inapplicable for inclusion in the task such as a flukicide or BZ/LEV combination product. Samples were included for analysis of drench efficacy if the pre-treatment flock egg count was ≥200 eggs per gram and the interval post-sampling was 10–14 days for BZ products, 4–7 days for LEV products and 14–18 days for ML products. These criteria reduced the number of valid tests to 369, 19.5% of all tests conducted. If the reduction post-treatment was ≥95% the treatment was considered effective. Only 51% of treatments were considered effective using this criterion. There was a significant difference in efficacy between the anthelmintic drug classes with BZ effective in only 30% of treatments, LEV effective in 52% of cases and ML effective in 76% of cases.

Conclusions

Gastrointestinal nematode anthelmintic treatments, as practiced on Irish farms, have a high failure rate. There was a significant difference between the efficacies of the anthelmintic classes with BZ the least effective and ML the most effective.     

Methods to determine resistance to anthelmintics when continuing larval development occurs

The in vivo faecal egg count reduction test (FECRT) is the most commonly used test to detect anthelmintic resistance (AR) in gastrointestinal nematodes (GIN) of ruminants in pasture based systems. However, there are several variations on the method, some more appropriate than others in specific circumstances. While in some cases labour and time can be saved by just collecting post-drench faecal worm egg counts (FEC) of treatment groups with controls, or pre- and post-drench FEC of a treatment group with no controls, there are circumstances when pre- and post-drench FEC of an untreated control group as well as from the treatment groups are necessary. Computer simulation techniques were used to determine the most appropriate of several methods for calculating AR when there is continuing larval development during the testing period, as often occurs when anthelmintic treatments against genera of GIN with high biotic potential or high re-infection rates, such as Haemonchus contortus of sheep and Cooperia punctata of cattle, are less than 100% efficacious. Three field FECRT experimental designs were investigated: (I) post-drench FEC of treatment and controls groups, (II) pre- and post-drench FEC of a treatment group only and (III) pre- and post-drench FEC of treatment and control groups.To investigate the performance of methods of indicating AR for each of these designs, simulated animal FEC were generated from negative binominal distributions with subsequent sampling from the binomial distributions to account for drench effect, with varying parameters for worm burden, larval development and drench resistance. Calculations of percent reductions and confidence limits were based on those of the Standing Committee for Agriculture (SCA) guidelines. For the two field methods with pre-drench FEC, confidence limits were also determined from cumulative inverse Beta distributions of FEC, for eggs per gram (epg) and the number of eggs counted at detection levels of 50 and 25. Two rules for determining AR: (1) %reduction (%R) < 95% and lower confidence limit <90%; and (2) upper confidence limit <95%, were also assessed. For each combination of worm burden, larval development and drench resistance parameters, 1000 simulations were run to determine the number of times the theoretical percent reduction fell within the estimated confidence limits and the number of times resistance would have been declared.When continuing larval development occurs during the testing period of the FECRT, the simulations showed AR should be calculated from pre- and post-drench worm egg counts of an untreated control group as well as from the treatment group. If the widely used resistance rule 1 is used to assess resistance, rule 2 should also be applied, especially when %R is in the range 90 to 95% and resistance is suspected.     

数种驱虫药和化学药物对体内外鸡蛔虫产卵和虫卵发育的影响

为鉴定男性不育药和有些驱虫药对寄生蠕虫的不育作用,对培养在试管内的鸡蛔虫和卵胚,以及人工感染蛔虫的鸡分别进行了体内和体外试验。棉酚和硝基呋喃类的呋喃唑酮、呋喃西林、S_(73031)和S_(72055)均无抑制产卵和卵胚发育的作用,苯并咪唑类的丙硫咪唑、甲苯咪唑和噻苯咪唑以及吩噻嗪均有抑制产卵和卵胚发育的作用,它们的EMIC_(50)分别为0.006,1.326,0.029和0.052ppm。抑制产卵所需浓度远大于抑制卵胚发育所需浓度。吡喹酮和硫双二氯酚也不具有抑制产卵和卵胚成熟的作用。放线菌酮仅对试管内蛔虫有抑制产卵作用,但对体内蛔虫却无影响。     

Anthelmintic resistance in non-strongylid parasites of horses

Since 2002, selected populations of Parascaris equorum in several countries have been reported to survive treatment with macrocyclic lactone (M/L) anthelmintics. Clinical treatment failures are characterized by negligible fecal egg count reduction, but M/L resistance has been confirmed in ascarids by controlled efficacy testing. Resistance was selected by current parasite control practices for foals, which often include exclusive and excessively frequent use of M/L dewormers, thereby minimizing refugia within the host and in the environment. Chemical control of M/L-resistant isolates can be accomplished with pyrimidine and/or benzimidazole anthelmintics, but a few M/L-resistant populations have recently exhibited resistance to pyrantel pamoate as well. Some specimens of Oxyuris equi regularly survive treatment with macrocyclic lactones, but it is uncertain whether this constitutes resistance or merely confirms the incomplete oxyuricidal efficacy of virtually all broad spectrum equine anthelmintics. Variations in other biological parameters of Oxyuris and Parascaris, specifically atypical infection of older hosts and shorter prepatent periods, have been reported anecdotally. These changes may represent genetic modifications that have evolved in parallel with resistance as a result of anthelmintic selection pressure.     

The in vitro diagnosis of anthelmintic resistance in cyathostomins

Cyathostomins are the primary parasitic pathogens of equids. For over 40 years, these nematodes have been controlled using broad spectrum anthelmintics. Three classes of anthelmintic are currently available for this use but, unfortunately, resistance to each of these has now been recorded in cyathostomin populations. As part of an optimal strategy to control cyathostomin infections in the field, it will be important to identify drug-resistant worms at as early a stage as possible. This objective needs to be supported by methodologies that will allow the accurate comparison of anthelmintic resistance in different nematode populations. At present, the faecal egg count reduction test is considered the most suitable method for initial screening for anthelmintic resistance in equine nematode populations. However, in its current state, this test lacks sensitivity. It is also costly and time-consuming to perform. Laboratory-based techniques, such as the egg hatch assay, larval development assay, larval migration inhibition assay and the larval feeding inhibition assay offer alternative options for assessing anthelmintic resistance in nematode populations. All of these tests have been investigated for their utility in measuring drug resistance in sheep nematode populations and some have proven useful. The egg hatch assay, larval development assay and larval migration inhibition assay have been investigated for use in measuring levels of drug resistance in equine nematode populations. However, at best, the results obtained thus far indicate that these tests require further refinement.     

Advances in laboratory diagnosis of parasitic infections of sheep

Parasitic infections constitute an important group of diseases in sheep concerning the health status, welfare and productivity. On a global scale, there are considerable differences concerning the epidemiological situation with respect of the various parasite species. However, there are also numerous species, which occur on all continents and, potentially, in every country. Accordingly, the present review aims to providing an overview about the recent developments in methods and technologies for the laboratory diagnosis of parasite infections in sheep. Following in principle a systematic order the review encompasses publications addressing the diagnosis of helminthes (i.e., trematodes, cestodes and nematodes) and arthropod species. New approaches using conventional (e.g., microscopic), immunological and molecular techniques are being considered. The diagnosis of anthelmintic resistance is highlighted separately, due to its significant importance. The review ends with an outlook into the future by discussing most recent technological advances, which might become of use for the diagnosis of parasite infections in sheep in the future.     

Resistance to a triple-combination anthelmintic in Trichostrongylus spp. on a commercial sheep farm in New Zealand

Abstract

AIM: To evaluate resistance to anthelmintics containing abamectin, levamisole, and oxfendazole (AB-LEV-OX), derquantal and abamectin (DEQ-AB), moxidectin, and monepantel in naturally acquired gastrointestinal nematodes present on a sheep farm.

METHODS: Faecal nematode egg count reduction tests (FECRT) were carried out on lambs that were approximately 7 months-old and infected with naturally acquired nematodes. Lambs were randomly allocated to one of five groups (n=15 per group): treatment with 2?mg/kg derquantel and 0.2?mg/kg abamectin; 0.2?mg/kg abamectin, 8?mg/kg levamisole HCl and 4.5?mg/kg oxfendazole; 2.5?mg/kg monepantel; 0.2?mg/kg moxidectin, or no treatment. Post-treatment samples were collected 12 days later. Abomasa and small intestines were collected from two slaughtered lambs from each of the DEQ-AB, AB-LEV-OX, moxidectin and control groups 15 days after treatment, for nematode counting.

RESULTS: The FECRT demonstrated that efficacy was 90.3 (95% CI=84.2–94.1)% for AB-LEV-OX, 54.5 (95% CI=28.4–71.1)% for moxidectin, 99.2 (95% CI=97.4–99.8)% for DEQ-AB and 100% for monepantel, across all genera. For Trichostrongylus spp. efficacy was 85.5% for AB-LEV-OX and 46.7% for moxidectin. Haemonchus spp. were fully susceptible to all treatments. Post-treatment nematode counts indicated that the resistant Trichostrongylus spp. were from the small intestine.

CONCLUSIONS: Anthelmintic resistance to both AB-LEV-OX and moxidectin was present in the Trichostrongylus genus on a commercial sheep farm. Monepantel and DEQ-AB were both effective against Trichostrongylus spp. based on FECRT results.

CLINICAL RELEVANCE: This finding of resistance to an AB-LEV-OX triple-combination anthelmintic in the Trichostrongylus genus in sheep in New Zealand further limits anthelmintic treatment options available, and calls into question whether this combination is suitable for use as a quarantine treatment.     

The response of vaccinated rams to experimental challenge using Brucella ovis

One hundred and thirty eight rams were allocated to four experimental groups. An inactivated Br. ovis vaccine was administered either once by the intraperitoneal route (1 i/p), twice by the intraperitoneal route (2 i/p), or twice by the subcutaneous route (2 s/c), and the last group was left unvaccinated. They were then challenged by the intravenous inoculation of between 123 and 1.23 × 10⁸ live Br. ovis bacteria. The number of rams that succumbed to infection within the four groups was 4/35 (11%) for the 2 s/c, 7/33 (21%) for the 2 i/p, 9/35 (26%) for the 1 i/p and 18/35 (51%) for the unvaccinated rams. Vaccination reduced the number of rams that succumbed to experimental challenge and although there were differences between the vaccinated groups, these were not statistically significant.

Following challenge, unvaccinated rams were the first to excrete Br. ovis in their semen; three weeks following inoculation. Next, those vaccinated by either one or two doses by the intraperitoneal technique began to excrete the organism (five weeks) and then finally those rams vaccinated twice by the subcutaneous route (seven weeks).