Search

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.

†‡ § # Joint last authors. Summary Reasons for performing study: Anthelmintic resistance in equine gastrointestinal nematodes is a threat to equine health and welfare. Detailed knowledge of anthelmintic use and parasite control methods is a prerequisite to identification of potential risk factors for resistance. Objectives: To identify parasite control practices employed by equine owners in Scotland and investigate management factors associated with anthelmintic resistance. Study design: Questionnaire study of equine parasite control in Scotland. Methods: Questionnaires were available electronically, distributed at a conference and mailed to clients. Key areas explored included general background, grazing management, anthelmintic treatment practices and use of diagnostic tests. Results: A total of 193 responses detailing information on parasite control programmes of 993 equids were analysed. Moxidectin (MOX) and ivermectin or related combination products were the most commonly administered anthelmintics in the preceding 12 months. Treatments licensed for use against cyathostomin encysted larvae and tapeworms were administered by 80% and 90% of respondents, respectively. This was often achieved through indiscriminate use of MOX and MOX-praziquantel products. Faecal egg count (FEC) analysis had been performed on 62% of yards and regular use of FECs reduced annual anthelmintic treatment frequency. Veterinarians had the greatest influence on control practices. While 40% of respondents believed that they practised targeted dosing, this was not associated with delaying treatment beyond the egg reappearance period of the anthelmintic used. Conclusions: Responses indicated increasing veterinary involvement and use of FECs. The majority of respondents administered anthelmintics licensed against cyathostomin encysted larvae and tapeworms. However, responses suggested that owners did not understand the definition of ‘targeted’ dosing regimens. Potential relevance: The high frequency of MOX use represents a potential risk factor for macrocyclic lactone resistance. As veterinarians were the most influential factor in anthelmintic choice, awareness of macrocyclic lactone resistance and potential risk factors for its development and spread should be incorporated into client advice.

Summary Reasons for performing study Cyathostomins comprise 50 helminth species, considered the most problematic equine endoparasites. Three classes of anthelmintic are currently licensed for their control, namely the benzimidazoles (fenbendazole), tetrahydropyrimidines (pyrantel) and macrocyclic lactones (ivermectin and moxidectin). Anthelmintic resistance in cyathostomins is common. With no new classes expected in the near future, it is essential to determine the efficacy of the available anthelmintics to inform future control programmes. Objectives To determine the efficacy of all 3 anthelmintic classes against strongyles in equids on livery yards in east and central Scotland. Study design Anthelmintic efficacy testing using the faecal egg count reduction test (FECRT). Methods FECRTs were performed on equids with initial strongyle faecal egg counts (FECs) of ≥50 eggs per gram. Efficacy was determined by comparing pretreatment (Day 0) and 14 days post treatment (Day 14) FECs. The following chemicals were tested: fenbendazole, pyrantel, ivermectin and moxidectin. Group arithmetic mean FECR of >90% for fenbendazole and pyrantel, and >95% for ivermectin and moxidectin, represented efficacy, whereas lower mean FECR indicated potential resistance. Results A total of 447 FECRTs were performed on 15 yards, as follows (the numbers in parentheses represent the number of yards each anthelmintic was tested on): 55 equids (7 yards) fenbendazole, 111 (8 yards) pyrantel, 163 (13 yards) ivermectin and 118 (10 yards) moxidectin. Fenbendazole resistance was documented on all yards (range of mean FECR, 15.8–83.4%), whereas pyrantel (90.4–99.6%), ivermectin (99.5–100%) and moxidectin (99.4–100%) treatment had acceptable efficacy. Conclusions Reduced efficacy of fenbendazole was widespread, whereas >90% efficacy was found after pyrantel, and >95% efficacy after ivermectin and moxidectin. Overall, efficacies were higher than reported previously in Europe and the USA, potentially reflecting differences in management and anthelmintic use on the yards surveyed. Potential relevance The use of fenbendazole for strongyle control in Scotland should be questioned. Targeted use of pyrantel should be encouraged to reduce reliance on macrocyclic lactones. Further work to correlate management practices with the presence of anthelmintic resistance is warranted. The Summary is available in Chinese – see Supporting information.

Summary Roundworms and flatworms that affect donkeys can cause disease. All common helminth parasites that affect horses also infect donkeys, so animals that co-graze can act as a source of infection for either species. Of the gastrointestinal nematodes, those belonging to the cyathostomin (small strongyle) group are the most problematic in UK donkeys. Most grazing animals are exposed to these parasites and some animals will be infected all of their lives. Control is threatened by anthelmintic resistance: resistance to all 3 available anthelmintic classes has now been recorded in UK donkeys. The lungworm, Dictyocaulus arnfieldi, is also problematical, particularly when donkeys co-graze with horses. Mature horses are not permissive hosts to the full life cycle of this parasite, but develop clinical signs on infection. In contrast, donkeys are permissive hosts without displaying overt clinical signs and act as a source of infection to co-grazing horses. Donkeys are also susceptible to the fluke, Fasciola hepatica. This flatworm can be transmitted, via snails and the environment, from ruminants. As with cyathostomins, anthelmintic resistance is increasing in fluke populations in the UK. A number of the anthelmintic products available for horses do not have a licence for use in donkeys, and this complicates the design of parasite control programmes. As no new equine anthelmintic classes appear to be near market, it is important that the efficacy of currently effective drugs is maintained. It is important that strategies are used that attempt to preserve anthelmintic efficacy. These strategies should be based on the concept that the proportion of worms in a population not exposed to anthelmintic at each treatment act as a source of ‘refugia’. The latter is an important factor in the rate at which resistance develops. Thus, it is imperative that parasite control programmes take into account the need to balance therapy to control helminth-associated disease with the requirement to preserve anthelmintic effectiveness.

Summary Reasons for performing study: Improved education of veterinarians and equine owners/managers is essential in implementing parasite control strategies that are less reliant on chemicals. Methods: This questionnaire study, conducted on 61 UK Thoroughbred (TB) establishments during 2009 and 2010, was designed to obtain an understanding of current helminth control practices on studs. To our knowledge, this is the first occasion that statements obtained from TB studs via questionnaire have been supported by statistical analysis. Results: Despite many respondents indicating high levels of concern regarding anthelmintic resistance, 56% of these establishments that received visiting equines co-grazed these animals with permanent stock and 25% of respondents), indicates that many stud owners/managers are not aware of the strong risk factors for the development of anthelmintic resistance. Few studs had conducted faecal egg count (FEC) analysis in the past and only 22% indicated that they considered this form of analysis beneficial in determining anthelmintic choice. Conclusions and potential relevance: The challenge now is to convince stud owners/managers to deviate from their current practices to control strategies that are more likely to preserve anthelmintic efficacy. Veterinarians need to get more involved in implementing these control strategies, with better emphasis placed on the role of diagnostic tests in facilitating targeted treatments and in investigating anthelmintic sensitivity in the associated nematode populations.

Summary Horses worldwide are exposed to a complex mixture of intestinal parasitic helminths. When burdens are high, these parasites can seriously compromise health and welfare. Some helminth species have an extremely high prevalence and are difficult to control, not least because there is a limited understanding of their most basic biology. Furthermore, levels of resistance to some of the commonly used anthelmintics are widespread and increasing. The cyathostomins are the most common nematode species affecting equids worldwide. Within this group of parasites are more than 50 different species. Until recent research activities, little was known about the contribution that individual species make to clinical disease, parasite epidemiology and anthelmintic resistance. This review describes some of the recent research advances in the understanding of cyathostomins in these areas. As part of the research effort, molecular tools were developed to facilitate identification of the non-parasitic stages of cyathostomins. These tools have proved invaluable in the investigation of the relative contributions that individual species make to the pathology and epidemiology of mixed infections. At the more applied level, research has also progressed in the development of a diagnostic test that will allow numbers of cyathostomin encysted larvae to be estimated. This test utilises cyathostomin-specific serum antibody responses as markers of infection. As anthelmintic resistance will be the major constraint on parasite control in future, researchers are actively investigating mechanisms of drug resistance and how to improve the detection of resistance in the field. Recent developments in these areas are also outlined.

SUMMARY Groups of yearling sheep, which had been trickle infected with Teladorsagia circumcincta for 8 weeks and then drenched, were challenged with 50 000 T. circumcincta larvae together with groups of worm-free controls. Fewer parasites and a greater proportion of early fourth stage larvae were recovered from previously infected sheep compared to controls. Worm loss and arrested development were evident by 5 days after challenge whereas growth retardation of developing worms was observed by day 10. In the previously infected sheep a secondary IgA response was observed in the efferent gastric lymph from 5 days post-infection. Western blot analysis showed the lymph IgA to be predominantly dimeric and nonsecretory in nature and that the somatic antigens recognized were predominantly in the 100‐250 kDa range. The concentration of IgA in lymph was always higher than in blood and in the previously infected sheep increased fivefold 8 days post-challenge in contrast to blood where IgA levels were unchanged. The timing of the response suggested that it occurred too late to have been the cause of worm loss or arrested development, though it may have retarded the growth of developing parasites .

Tumour necrosis factor alpha (TNF-alpha) is a pro-inflammatory cytokine that plays a crucial role in the regulation of inflammatory and immune responses. In all vertebrate species the genes encoding TNF-alpha are located within the major histocompatability complex. In the horse TNF-alpha has been ascribed a role in a variety of important disease processes. Previously two single nucleotide polymorphisms (SNPs) have been reported within the 5' un-translated region of the equine TNF-alpha gene. We have examined the equine TNF-alpha promoter region further for additional SNPs by analysing DNA from 131 horses (Equus caballus), 19 donkeys (E. asinus), 2 Grant's zebras (E. burchellii boehmi) and one onager (E. hemionus). Two further SNPs were identified at nucleotide positions 24 (T/G) and 452 (T/C) relative to the first nucleotide of the 522 bp polymerase chain reaction product. A sequence variant at position 51 was observed between equidae. SNaPSHOTtrade mark genotyping assays for these and the two previously reported SNPs were performed on 457 horses comprising seven different breeds and 23 donkeys to determine the gene frequencies. SNP frequencies varied considerably between different horse breeds and also between the equine species. In total, nine different TNF-alpha promoter SNP haplotypes and their frequencies were established amongst the various equidae examined, with some haplotypes being found only in horses and others only in donkeys or zebras. The haplotype frequencies observed varied greatly between different horse breeds. Such haplotypes may relate to levels of TNF-alpha production and disease susceptibility and further investigation is required to identify associations between particular haplotypes and altered risk of disease.

Reasons for performing study Anthelmintic resistance is a global problem and constitutes a major threat to the welfare of equids worldwide. The cyathostomins are the most numerous and pathogenic gastrointestinal nematode (GIN) of equids in the developed world. Cyathostomins show widespread resistance to 2 out of 3 of the major classes of anthelmintic and recently there are reports of reduced efficacy to the potent macrocyclic lactones (MLs). None of the 3 novel classes of anthelmintic that have emerged in the last decade are licensed for use in equids. The cysteine proteases (CPs) are plant proteins that have shown potent activity against GINs in vivo in sheep and pigs. Objectives This study aimed to evaluate the anthelmintic effect of the CP papain on cyathostomins in vitro using the egg hatch assay (EHA) and larval migration inhibition test (LMIT). Methods Samples of cyathostomin eggs and third stage larvae were collected and cultured from a population of equids that have recently shown reduced ML efficacy in vivo. The EHA and LMIT were performed on repeated samples with increasing concentrations of papain. Dose–response curves were plotted and PROBIT analysis performed on the data to give EC-50 values (concentration that gives 50% of the maximal response). Results Papain caused a dose dependent inhibition of both egg hatch and larval migration. The EC-50 values were 2 μmol/l and 100 μmol/l in the EHA and LMIT respectively, indicating a more potent effect on egg hatch. Conclusions The CP papain shows potent anthelmintic activity against cyathostomins in vitro. Good evidence of anthelmintic effect against GINs in other host species is supportive of its potential use in equids. Further work is indicated to evaluate safety and in vivo efficacy. Ethical animal research: The study was approved by the University of Liverpool and Donkey Sanctuary Ethics Committee. Explicit owner informed consent for participation in this study was not stated. Sources of funding: The Donkey Sanctuary, University of Liverpool – Institute of Infection and Global Health. Competing interests: None.

Fourteen dental cysts containing intra-epithelial hyaline bodies were stained for keratin, Factor VIII-related antigen, haemoglobin and fibrinogen using immunoperoxidase methods. Although the hyaline bodies were negative for all tested antigens fibrinogen was detected in the cores of some circular and polycyclic forms. These observations do not provide support for the proposed keratin, capillary endothelium or erythrocyte nature of these structures. The core staining for fibrinogen may support the theory that hyaline bodies are produced by a cellular reaction to extravasated serum.

Attempts to measure the electrification produced by the melting of ice and snow, known as the Dinger-Gunn effect, have been made with three different experimental arrangements. In one of these snow crystals were grown and melted under very clean conditions inside a diffusion cloud chamber. The results of the three experiments are consistent in failing to detect a separation of charge and in indicating that any charge produced was < 10−2 e.s.u./g or two orders of magnitude smaller than reported by Dinger and Gunn. This result was unaffected by varying the purity of the ice, its air content and the rates of freezing and melting, and the pH value of the water. Accordingly, it is concluded that charging associated with the melting of snow or hail is unlikely to be an important feature in the electrification of clouds and precipitation.