United Kingdom

The evaluation of ethnoveterinary medicines for treating gastrointestinal nematodes in working equids

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Date presented: 
Wednesday 2 July 2014
Abstract

Introduction: Herbal medicines have been used in human and animal medicine for centuries to treat parasitic diseases; few examples have been investigated for genuine anti-parasitic activity. In developing countries access to effective anthelmintic treatment for livestock is often limited by cost, availability and variable quality. Reports of resistance to benzimadazoles in ruminants in Ethiopia serve as a warning that anthelmintic resistance may also be an emerging problem [1,2]. In light of these issues there is increasing interest in plant remedies as alternatives to synthetic anthelmintics. This study used a participatory rural appraisal (PRA) to identify plants with potential anthelmintic activity in the Oromia region of Ethiopia; five plant extracts were shortlisted and tested for efficacy against cyathostomins using in vitro assays. Current attitudes to ethnoveterinary medicine were discussed.

Methods: Focus group discussions with 29 groups of donkey owners from the Oromia region of Ethiopia explored the use of plants to treat GI parasites in livestock. Current attitudes to herbal medicines were discussed and recorded using thematic analysis. Plants of interest were collected and identified at the National Herbarium, Addis Ababa, Ethiopia. Plants were shortlisted for in vitro tests based on four criteria; ranking in the PRA, supportive literature, no evidence of toxicity and availability. Hydro-alcoholic extraction of dried plant material from shortlisted species was performed. The efficacy of extracts was evaluated in the egg hatch assay (EHA) using cyathostomin eggs recovered from the faeces of donkeys at the Donkey Sanctuary, UK. Dose response curves were produced and ED-50 values calculated using probit analysis.

Results: The focus groups identified 21 plants used as anthelmintics in livestock. A general move away from traditional medicines in the younger generation was observed, although when asked if they would use plants in future many would consider this if they had been tested scientifically and were approved by professionals. The five plants shortlisted for in vitro analysis were Acacia nilotica, Cucumis prophetarum, Rumex abysinnicus, Vernonia amygdalinia and Withania somnifera. Three showed efficacy in the EHA; Acacia nilotica, Cucumis prophetarum and Rumex abysinnicus , with EC-50 values of 0.7, 1.1 and 1.3mg/ml respectively.

Conclusion: Three out of five of the plants identified in the PRA showed efficacy in vitro suggesting that plant remedies used by livestock owners in the Oromia region of Ethiopia may contain compounds with genuine anthelmintic activity. Evaluation of current attitudes suggests that plant remedies are not used unless there is no other option, but that they would be considered should scientific evidence of efficacy and safety be presented to them by animal health professionals [3]. It is therefore essential that a randomised controlled trial is used to verify whether in vitro anthelmintic activity can be translated in vivo and thus whether the plants identified in this study have potential as safe alternatives to synthetic anthelmintic drugs. This study has highlighted that local practices pertaining to the health of working equids are a rich source of information that may help to inform sustainable and effective treatment strategies in future.

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The P-glycoprotein inhibitor ketoconazole causes a reversion to ivermectin sensitivity in cyathostomins in vitro

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Laura Peachey, Jacqui. B. Matthews, Gina L. Pinchbeck, Faith A. Burden, Nikki Stradling, Jane Hodgkinson. The P-glycoprotein inhibitor ketoconazole causes a reversion to ivermectin sensitivity in cyathostomins in vitro. Presented at British Society for Parasitology Spring Meeting.

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Date presented: 
Monday 7 April 2014
Abstract

Anthelmintic resistance is a major veterinary and public health issue globally, of most concern is the level of resistance to the macrocyclic lactones. Recent studies have identified a role in resistance for the ATP binding cassette (ABC) drug transporters, P-glycoproteins (P-gps). This study demonstrates the effect of the P-gp inhibitor ketoconazole on the efficacy of ivermectin (IVM) against equid cyathostomin larvae using the larval migration inhibition test (LMIT). Third stage cyathostomin larvae (L3) were cultured from two populations; 1) with recent history of IVM resistance in vivo and 2) naive to anthelmintic exposure. The sensitivity to IVM in each group (n=8) was characterised using the LMIT. The IVM LMIT was repeated for each sample with and without the addition of 10µM ketoconazole. Probit analysis was performed on grouped data from each population to give LC-50 values. The LC-50 value for IVM in Populations 1 and 2 was 4.9 and 2.4µg/ml respectively indicating that Population 1 has a resistant phenotype in comparison to Population 2. Addition of 10µM ketoconazole to IVM in Population 1 caused a drop in LC-50 value from 5.8 to 1.6µg/ml. In Population 2 the effect of the addition of ketoconazole was negligible (1.1 to 0.9µg/ml). This study demonstrates that the P-gp inhibitor ketoconazole causes reversion to a sensitive phenotype in IVM-resistant cyathostomins, inferring that P-gps play a role in their resistance to IVM. This work will be corroborated by investigation into P-gp genes and their expression in cyathostomins.

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Quantifying the effects of individual animal characteristics and climatological factors on faecal worm egg count shedding in donkeys

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Christopher J Corbett, Sandy Love, Giles T. Innocent, James McKendrick, Jacqui. B. Matthews, Louise Matthews, Matthew Denwood, Faith A. Burden. Quantifying the effects of individual animal characteristics and climatological factors on faecal worm egg count shedding in donkeys. Presented at British Society for Parasitology Spring Meeting.

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Date presented: 
Monday 7 April 2014
Abstract

Cyathostomins, the predominant parasitic nematodes of equids, have developed varying degrees of resistance to all three classes of anthelmintic licensed for use in horses. It is essential that the effectiveness of alternative methods of control for these pathogens are quantified, including incorporating climatic data and the commonly advocated practice of removal of faeces from pasture. Here, we obtained monthly faecal worm egg counts (FWEC, n=4,460 individual counts) from 803 donkeys based at The Donkey Sanctuary (Devon, UK). The dataset also included age, sex, field, FWEC history and previous anthelmintic administrations in each individual, as well as the pasture hygiene management method applied in the field where the donkey was grazed. FWEC were analysed alongside local climatic data using a generalised linear mixed model to assess associations between these variables and each observed monthly FWEC. The preferred model was identified using a model selection algorithm based on penalised likelihoods, and associated a 2.1% decrease in FWEC per day with air frost two calendar months ago (p<0.001) and a 38% lower FWEC in groups with twice weekly manual faecal removal compared to those with no faecal removal (p=0.004). Other weather effects, both alone and as interaction terms with the average FWEC of the field were included in the model, alongside individual FWEC history with anthelmintic administration as interaction terms and date as a single term. Our study identifies factors that may be useful as part of on-going predictive modelling based methods of improving targeted selective therapy.

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Chronic pleuropulmonary fibrosis and elastosis of aged donkeys - similarities to human pleuroparenchymal fibroelastosis (PPFE)

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Amy Miele, K Dhaliwal, Nicole du Toit, J Murchison, C Dhaliwal, Harriet Brooks, Sionagh H. Smith, N Hirani, T Schwarz, C Haslett, W Wallace, Bruce McGorum. March 2014. Chronic pleuropulmonary fibrosis and elastosis of aged donkeys - similarities to human pleuroparenchymal fibroelastosis (PPFE). Chest.

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Publication date: 
6 March 2014
Journal: 
Chest
DOI number: 
10.1378/chest.13-1306
Abstract

BACKGROUND:
Donkey Pulmonary Fibrosis (DPF) is a spontaneous syndrome of aged donkeys with high prevalence (35%). No previous detailed characterisation of DPF has been performed. We sought to determine the similarities of DPF to recognised patterns of human pulmonary fibrosis.

METHODS:
Whole lungs were collected from 32 aged donkeys at routine necropsy. Gross examination revealed pulmonary fibrosis in 19 donkeys (DPF cases), while 13 (controls) had grossly normal lungs. Eighteen whole inflated ex vivo lungs (11 DPF, 7 controls) were imaged with high resolution computed tomography (HRCT), while the remainder were sectioned and photographed. Tissue samples were collected from all lungs for histopathological evaluation using a standardised protocol. HRCT images and histology sections were reviewed independently and blindly. Lung tissue was analysed for herpes virus, fungal hyphae, mycobacteria and dust content.

RESULTS:
Ten of 19 DPF lungs were categorised as being 'consistent with' pleuroparenchymal fibroelastosis (PPFE) according to previously defined histological and imaging criteria. All 10 PPFE-like lungs had marked pleural and subpleural fibrosis, predominantly within the upper lung zone, with accompanying intra-alveolar fibrosis and elastosis. Asinine herpesvirus (AsHV) was ubiquitously expressed within control and DPF lung tissue. No other aetiological agents were identified.

CONCLUSIONS:
Many cases of DPF share key pathological and imaging features with human PPFE, a rare interstitial pneumonia. Consequently, further study of DPF may help elucidate the aetiopathogenesis of human PPFE.

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The P-glycoprotein inhibitor ketoconazole causes a reversion to sensitivity in ivermectin resistant cyathostomins in vitro

Citation

Laura Peachey, Jacqui. B. Matthews, Gina L. Pinchbeck, Faith A. Burden, Jane Hodgkinson. The P-glycoprotein inhibitor ketoconazole causes a reversion to sensitivity in ivermectin resistant cyathostomins in vitro. Presented at Anthelmintics: From Discovery to Resistance. (5 February - 7 February 2014). San Francisco, USA.

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Date presented: 
Thursday 6 February 2014
Abstract

Anthelmintic resistance is a growing problem in both the developed and developing world; of most concern is the level of resistance detected against the potent macrocylic lactone (ML) anthelmintics. To identify and target a common mechanism of resistance to anthelmintics would allow potential modification of existing drugs, and may even enable the prediction and prevention of the development of resistance to novel drugs. There is a growing body of evidence that P-glycoproteins (P-gps) are involved in resistance to the MLs in many parasitic nematodes of humans and veterinary species (Ardelli et al, 2011). P-gps belong to class two of the ATP binding cassette (ABC) transporter protein superfamily; they are responsible for the active removal of xenobiotic compounds from cells. The cyathostomins are gastrointestinal nematodes of equids that cause significant pathology. Recently resistance to MLs has been described in cyathostomins (Molento et al, 2008), its mechanisms have not yet been elucidated.

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The effectiveness of faecal removal methods of pasture management to control the cyathostomin burden of donkeys

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Christopher J Corbett, Sandy Love, Anna Moore, Faith A. Burden, Jacqui. B. Matthews, Matthew Denwood. January 2014. The effectiveness of faecal removal methods of pasture management to control the cyathostomin burden of donkeys. Parasites and Vectors. 7:48.

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Publication date: 
24 January 2014
Volume: 
7
Issue: 
48
DOI number: 
doi:10.1186/1756-3305-7-48
Abstract

Background: The level of anthelmintic resistance within some cyathostomin parasite populations has increased to the level where sole reliance on anthelmintic-based control protocols is not possible. Management-based nematode control methods, including removal of faeces from pasture, are widely recommended for use in association with a reduction in anthelmintic use to reduce selection pressure for drug resistance; however, very little work has been performed to quantitatively assess the effectiveness of such methods.

Methods: We analysed data obtained from 345 donkeys at The Donkey Sanctuary (Devon, UK), managed under three different pasture management techniques, to investigate the effectiveness of faeces removal in strongyle control in equids. The management groups were as follows: no removal of faeces from pasture, manual, twice weekly removal of faeces from pasture and automatic, twice-weekly removal of faeces from pasture (using a mechanical pasture sweeper). From turn-out onto pasture in May, monthly faecal egg counts were obtained for each donkey and the dataset subjected to an auto regressive moving average model.

Results: There was little to no difference in faecal egg counts between the two methods of faecal removal; both resulted in significantly improved cyathostomin control compared to the results obtained from the donkeys that grazed pasture from which there was no faecal removal.

Conclusions: This study represents a valuable and unique assessment of the effectiveness of the removal of equine faeces from pasture, and provides an evidence base from which to advocate twice-weekly removal of faeces from pasture as an adjunct for equid nematode control. Widespread adoption of this practice could substantially reduce anthelmintic usage, and hence reduce selection pressure for nematode resistance to the currently effective anthelmintic products.

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Expression of PGP 9.5 by Enteric Neurons in Horses and Donkeys with and without Intestinal Disease

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Neil Hudson, G.T. Pearson, I.G. Mayhew, Christopher Proudman, Faith A. Burden, Constance Fintl. November 2013. Expression of PGP 9.5 by Enteric Neurons in Horses and Donkeys with and without Intestinal Disease. Journal of Comparative Pathology.

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Publication date: 
26 November 2013
DOI number: 
doi.org/10.1016/j.jcpa.2013.11.203
Abstract

Intestinal motility disorders are an important problem in horses and donkeys and this study was carried out in order to evaluate the enteric neurons in animals with and without intestinal disease. Surplus intestinal tissue samples were collected from 28 horses undergoing exploratory laparotomy for colic. In addition, surplus intestinal samples from 17 control horses were collected immediately following humane destruction for clinical conditions not relating to the intestinal tract. Similar samples were also collected during routine post-mortem examinations from 12 aged donkeys; six animals were humanely destroyed for conditions related to the intestinal tract, while the remaining six were humanely destroyed for other reasons including dental and orthopaedic diseases. Tissue samples were fixed in formalin and immunohistochemical labelling was performed targeting the enteric neurons using a polyclonal antibody specific for the neuronal marker PGP 9.5. The distribution and density of neuronal networks were assessed qualitatively and semiquantitatively. There was strong PGP 9.5 expression in both the horse and donkey samples and labelling was detected throughout the tissue sections. In both species, PGP 9.5-immunoreactive nerve fibres were detected in all layers of the intestinal tract, both in large and small intestinal samples. Networks of enteric neurons were present in the donkey with a similar distribution to that seen in the horse. There was no demonstrable difference in enteric neuronal density and distribution in the groups of animals with intestinal disease compared with those without, apart from two (out of 28) horses with intestinal disease that showed a marked reduction in PGP 9.5 immunoreactivity. Apart from these two animals, this total cohort analysis differs from some previously observed findings in horses with intestinal disease and may therefore reflect the different pathophysiological processes occurring in varying intestinal conditions resulting in colic both in the donkey and the horse.

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Donkey assisted therapy

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David Cook, Stephen Blakeway. Donkey assisted therapy. Presented at Donkey Welfare Symposium. (1 November 2013). California, USA.

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Date presented: 
Saturday 3 November 2012

Handling untrained and poorly trained donkeys

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Ben Hart. Handling untrained and poorly trained donkeys. Presented at Donkey Welfare Symposium. (1 November 2013). California, USA.

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Date presented: 
Friday 1 November 2013
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