anthelmintic

Papaya latex supernatant has a potent effect on the free-living stages of equid cyathostomins in vitro

Citation

Laura Peachey, Gina L. Pinchbeck, Jacqui. B. Matthews, Faith A. Burden, J.M. Behnke, Jane Hodgkinson. August 2016. Papaya latex supernatant has a potent effect on the free-living stages of equid cyathostomins in vitro. Veterinary Parasitology.

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Publication date: 
1 August 2016
DOI number: 
10.1016/j.vetpar.2016.07.036
Abstract

The control of equid gastrointestinal nematodes in developed countries, in particular the cyathostomins, is threatened by high levels of anthelmintic resistance. In recent years, there has been increasing interest in the evaluation of traditional ‘ethnoveterinary’ medicines as alternatives to chemical anthelmintics. The cysteine proteinases (CPs), a group of enzymes derived from fruits such as papaya (Carica papaya), pineapple (Ananas comosus) and figs (Ficus spp.), have shown good efficacy against adult stages of a range of parasitic nematodes, in vitro and in vivo. The efficacy of CPs against cyathostomins remains to be explored. In this study, the efficacy of a crude preparation of CPs, papaya latex supernatant (PLS), against the free-living stages of cyathostomins was evaluated using two in vitro tests, the egg hatch test (EHT) and the larval migration inhibition test (LMIT). It was demonstrated that PLS had a potent effect in the EHT, with EC-50 values in the range of 0.12-0.22 μM. At concentrations above 6.25 μM the eggs did not develop, below this concentration the L1 developed but they lost integrity of the cuticle upon hatching. These effects were inhibited by pre-incubation of PLS with the CP inhibitor L-trans-epoxysuccinyl-L-leucylamido-(4-guanidino butane) (E64), indicating that CPs were responsible for the anti-parasitic activity. A dose-dependent inhibition of migration of third stage larvae (L3) in the LMIT was demonstrated at higher concentrations of PLS, with EC-50 values in the range of 67.35-106.31 μM. Incubation of PLS with E64 prior to use in the LMIT did not reverse the anti-migratory effect, suggesting that CPs were not responsible for the reduced migration of cyathostomin L3 and that PLS also contains an additional active compound. This is the first report of PLS and/or CPs showing activity against the free-living stages of a parasitic helminth. In addition, it suggests that cyathostomins are highly sensitive to the effects of CPs and further evaluation of their efficacy against parasitic stages and in vivo are strongly indicated.

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Development of the larval migration inhibition test for comparative analysis of ivermectin sensitivity in cyathostomin populations

Citation

Claire McArthur, Ian G Handel, Ailie Robinson, Jane Hodgkinson, Barend MdeC Bronsvoort, Faith A. Burden, Ray Kaplan, Jacqui. B. Matthews. June 2015. Development of the larval migration inhibition test for comparative analysis of ivermectin sensitivity in cyathostomin populations. Veterinary Parasitology.

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Publication date: 
20 June 2015
DOI number: 
10.1016/j.vetpar.2015.06.019
Abstract

Cyathostomins are the most prevalent parasitic pathogens of equids worldwide. These nematodes have been controlled using broad-spectrum anthelmintics; however, cyathostomin resistance to each anthelmintic class has been reported and populations insensitive to more than one class are relatively commonplace. The faecal egg count reduction test (FECRT) is considered the most suitable method for screening anthelmintic sensitivity in horses, but is subject to variation and is relatively time-consuming to perform. Here, we describe a larval migration inhibition test (LMIT) to assess ivermectin (IVM) sensitivity in cyathostomin populations. This test measures the paralysing effect of IVM on the ability of third stage larvae (L3) to migrate through a pore mesh. When L3 from a single faecal sample were examined on multiple occasions, variation in migration was observed: this was associated with the length of time that the L3 had been stored before testing but the association was not significant. Half maximal effective concentration (EC50) values were then obtained for cyathostomin L3 from six populations of horses or donkeys that showed varying sensitivity to IVM in previous FECRTs. Larvae from populations indicated as IVM resistant by FECRT displayed significantly higher EC50 values in the LMIT than L3 from populations classified as IVM sensitive or L3 from populations that had not been previously exposed to IVM or had limited prior exposure. The analysis also showed that EC50 values obtained using L3 from animals in which IVM faecal egg count reduction (FECR) levels had been recorded as <95% were significantly higher than EC50 values obtained using L3 from animals for which FECR was measured as >95%. For one of the populations, time that had elapsed since IVM administration had an effect on the EC50 value obtained, with a longer time since treatment associated with lower EC50 values. These results indicate that the LMIT has value in discriminating IVM sensitivity amongst cyathostomin populations, but several factors were identified that need to be taken into account when executing the test and interpreting the derived data.

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A Potential Novel Anthelmintic? The Cysteine Proteases Show Potent Anthelmintic Activity Against Cyathostomins In Vitro

Citation

Laura Peachey, Jacqui. B. Matthews, Gina L. Pinchbeck, J. Benkhe, Faith A. Burden, Jane Hodgkinson. September 2014. A Potential Novel Anthelmintic? The Cysteine Proteases Show Potent Anthelmintic Activity Against Cyathostomins In Vitro. Equine Veterinary Journal. 46. 23.

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Publication details
Publication date: 
8 September 2014
Volume: 
46
Page numbers: 
23
DOI number: 
10.1111/evj.12323_52
Abstract

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.

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Clinical trial on the efficacy of moxidectin oral gel formulation on donkeys naturally infected by cyathostominae

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Presentation details
Date presented: 
Saturday 7 February 2015
Abstract

Donkeys and horses share several parasites including the small strongyles, Cyathostominae. Moxidectin (MOX), a compound of macrocyclic lactones, has a wide range of ecto and endoparasitic activity in many species. For horses, MOX is available as oral gel formulation that provides excellent and long-lasting efficacy against nematodes such as large and small strongyles. There is a paucity of data available on the efficacy of anthelmintics used in donkeys (Veneziano et al., 2011). Therapeutics, such as antiparasitic compounds, are often administered to donkeys on the basis of dosage and intervals recommended for horses, because very few drugs have donkey-specific label indications (Grosenbaugh et al., 2011). The objective of the present study was to evaluate the field efficacy and Egg Reappearance Period (ERP) of MOX oral gel up to 84 days at horse dose against natural infection of Cyathostominae in donkeys.

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Participatory study of medicinal plants used in the control of gastrointestinal parasites in donkeys in Eastern Shewa and Arsi zones of Oromia region, Ethiopia

Citation

Claire Scantlebury, Laura Peachey, Jane Hodgkinson, Jacqui. B. Matthews, Andrew F. Trawford, Mulugeta Getachew, Gebre Tefera, Gina L. Pinchbeck. September 2013. Participatory study of medicinal plants used in the control of gastrointestinal parasites in donkeys in Eastern Shewa and Arsi zones of Oromia region, Ethiopia. BMC Veterinary Research. 9:179. 1-12.

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Publication details
Publication date: 
1 September 2013
Volume: 
9
Issue: 
179
Page numbers: 
1-12
DOI number: 
doi:10.1186/1746-6148-9-179
Abstract

Background

Gastrointestinal nematode infections constitute a threat to the health and welfare of donkeys worldwide. Their primary means of control is via anthelmintic treatments; however, use of these drugs has constraints in developing countries, including cost, limited availability, access to cheaper generic forms of variable quality and potential anthelmintic resistance. As an alternative, bioactive plants have been proposed as an option to treat and control gastrointestinal helminths in donkeys. This study aimed to use participatory methodology to explore donkey owner knowledge, attitudes and beliefs relating to the use of plant-based treatments for gastrointestinal parasites of donkeys in Ethiopia.

Results

In focus groups, 22/29 groups stated they knew of plants used for the treatment of gastrointestinal parasites in donkeys. All groups volunteered plants that were used in cattle and/or small ruminants. In total, 21 plants were named by participants. ‘Koso’ (Hagenia abyssinica) ‘Grawa’ (Vernonia amygdalina) and a mixed roots and leaves preparation were the most frequently named plant preparations. ‘Enkoko’ (Embelia shimperi) and ‘a mixture of roots and leaves’ were ranked highly for effectiveness in donkeys. However, ‘Grawa’ and ‘Koso’ were the highest ranked when taking into account both the rank position and the number of groups ranking the plant.

Thematic analysis of participants’ current attitudes and beliefs surrounding traditional plant-based remedies for gastrointestinal parasites revealed that anthelmintics obtained from clinics were generally favoured due to their ease of administration and perceived higher effectiveness. There was doubt surrounding the effectiveness of some plant-based treatments, but there were also perceived advantages including their low cost, ease of cultivation and availability. However, plant-based treatments were considered a “past trend” and people favoured “modern” medicine, particularly among the younger generation.

Conclusions

There was extensive knowledge of plant-based treatments for gastrointestinal parasites in livestock in Ethiopia. In donkeys, Koso (Hagenia abyssinica), Grawa (Vernonia amygdalina), Enkoko (Embelia shimperi) and ‘mixed roots and leaves’ were the most frequently named and/or highest ranked plants with reported efficacy against gastrointestinal parasites. Further in vitro and in vivo investigation of these plants is now required to determine viable alternatives for the treatment and control of gastrointestinal parasites in Ethiopia.

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Drug Resistant Cyathostomins in Donkey Herds; Lessons in Management for All Equids

Citation

Andrew F. Trawford, Faith A. Burden. Drug Resistant Cyathostomins in Donkey Herds; Lessons in Management for All Equids. Presented at International Conference on Equine Infectious Diseases IX. (21 October - 26 October 2012). Kentucky, USA.

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Presentation details
Date presented: 
Sunday 23 October 2011
Abstract

The Donkey Sanctuary has over 2500 donkeys on its farms in the UK. Endoparasite control in these herds has been a major focus for the last 10 years. Monitoring of the herds has identified significant levels of anthelmintic resistance in the cyathostomins parasitising the donkey herds. Lack of efficacy of ivermectin, moxidectin, pyrantel and fenbendazole has been identified on multiple sites and in a number of cases treatment of donkeys harbouring such parasites is becoming increasingly challenging. Changes in management practices and approach to parasite treatment is discussed. Research to establish the extent of drug resistance and clinical significance of parasitism in donkeys is of primary concern; projects have focussed on identification of drug resistance, impacts of pasture management, validation of thresholds of faecal egg counts for treatment and alternatives to anthelmintics. Significant changes in management practices have been based upon results of these studies and new ideas and techniques are being developed. Treatment of all donkeys is carried out on the basis of a strongyle faecal egg count which is carried out on a 4-weekly basis throughout the year. Preliminary data and clinical experience has shown that increasing the FEC treatment threshold for healthy, mature donkeys to ≥ 1000 strongyle epg is sufficient to control clinical disease whilst reducing pasture contamination. Application of new treatment thresholds have reduced the reliance on chemical treatment. The mean number of anthelmintic treatments per year per animal was one in 2011; this is a significant reduction from previous dosing regimes where animals would be treated a minimum of four times per year. Animals are monitored frequently with problematic herds being 100% sampled every four weeks; such monitoring allows individualised treatment programmes and highlights reduced egg reappearance periods early. Reduced ERPs are investigated immediately with suspect animals undergoing a faecal egg count reduction test (FECRT) at the next treatment. Failed FECRTs trigger an alternative approach to treatment and further investigation. It is evident that some individual cases are becoming increasingly difficult to treat effectively; future trials of combinations of anthelmintics are being considered. Effective parasite control has also led to an increased emphasis on pasture management; dung is closely managed and carefully composted. Pasture is also rested in 6 month blocks with mixed species grazing instigated where possible. Good pasture management practices produce significant drops in pasture larval counts and is the primary mechanism used to reduce parasitism. Research and practical experience has emphasised the importance of good pasture management, treatment based upon faecal egg counts and routine assessment of efficacy of treatment. Researching and trialling higher dosing threshold has also led to significant decreases in the reliance of the organisation on anthelmintics with no recorded clinical effects. The lessons learnt from management of these large herds of donkeys may provide salient and important foundations for future equid herds that undoubtedly will experience similar problems in the future.

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