Kristopher Hughes

Type 2 diabetes mellitus with pancreatic β cell dysfunction in 3 horses confirmed with minimal model analysis.

REASONS FOR PERFORMING STUDY Type 2 diabetes mellitus (T2DM) is diagnosed rarely in equine practice although it may be under-recognised. A greater awareness of the condition and therapeutic considerations would be to the benefit of such cases presenting in practice. More investigation into the pharmacological management of these cases is needed. OBJECTIVES Three cases of diabetes mellitus were investigated using a specific test for insulin sensitivity and pancreatic beta cell function in order to define accurately and characterise the existence of T2DM in all 3 subjects. METHODS The insulin-modified frequently sampled i.v. glucose tolerance test was performed in each case and the data so obtained were subject to minimal model analysis of insulin-glucose dynamics. Cases were then monitored following treatment using a combination of dietary modification, metformin, glibenclamide and pergolide. RESULTS Marked insulin resistance was identified in each case and, furthermore, severe pancreatic beta cell dysfunction was present therefore classifying each case as end stage T2DM. Treatment was nevertheless associated with restoration of normoglycaemia in all cases. CONCLUSIONS T2DM in horses may be more common than generally considered.In some cases individuals may respond to therapy aimed at restoring insulin sensitivity and pancreatic function. Drugs used in other species for the treatment of T2DM have not yet been adequately tested in horses. POTENTIAL RELEVANCE T2DM should be considered as an important differential diagnosis in mature to elderly horses and ponies suffering from weight loss, polydipsia and polyuria. Clinicians should be encouraged to offer treatment and management advice when such cases are encountered.

Assessment of subclinical venous catheter-related diseases in horses and associated risk factors

A total of 102 horses that had a catheter introduced intravenously to facilitate treatment had the catheterised jugular vein and contralateral vein examined by ultrasound every 48 hours. Subclinical complications were defined by thrombus formation or thickening of the venous wall, and the data were analysed to establish risk factors for the development of these complications. The horses with a rectal temperature above 38·5°C when the catheter was introduced were four times more likely to develop complications, than the horses with a lower temperature. The administration of a NSAID while the catheter was in place reduced the risk of complications developing.

Evaluation of cytokine mRNA expression in bronchoalveolar lavage cells from horses with inflammatory airway disease

Inflammatory airway disease (IAD) is a common disorder of performance horses and is associated with poor performance and accumulation of mucus and inflammatory cells in lower airway secretions. Horses with IAD frequently have increased relative counts of neutrophils in bronchoalveolar lavage fluid (BALF); less commonly relative counts of eosinophils and/or mast cells may be increased. The aetiopathogenesis of IAD is unknown and may involve innate and/or acquired immune responses to various factors including respirable dust constituents, micro-organisms, noxious gases and unconditioned air. The molecular pathways and role of the immune system in the pathogenesis of IAD remain poorly defined and it is unknown whether polarised T cell responses occur in the disease, as have been reported to occur in equine recurrent airway obstruction and asthma in humans. Elucidating cytokine responses that develop in horses with IAD may allow a greater understanding of the possible aetiopathological pathway(s) involved and could contribute to development of novel treatments. We compared the mRNA expression of tumour necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-8, IL-13, IL-17 and IL-23 in cell pellets extracted from BALF of horses with IAD (n=21) and horses free of respiratory tract disease (n=17). Horses with IAD had significantly increased levels of TNF-α, IL-1β and IL-23 mRNA; no significant differences in the other cytokine mRNAs were detected. The results of this study indicate that IAD of horses is associated with increased mRNA expression of pro-inflammatory cytokines in BALF cells, which may reflect stimulation of the innate immune responses to inhaled antigens. There was no evidence of a polarised T-cell cytokine response suggesting hypersensitivity responses may not be involved in the aetiopathogenesis of IAD.

Investigation of rhythms of secretion and repeatability of plasma adrenocorticotropic hormone concentrations in healthy horses and horses with pituitary pars intermedia dysfunction

REASONS FOR PERFORMING STUDY There is little published information on whether measurement of plasma ACTH concentration at a single timepoint is a repeatable indicator of pituitary pars intermedia dysfunction (PPID). OBJECTIVES To determine whether ultradian or circadian fluctuations in ACTH production influence plasma ACTH concentration in normal horses and horses with PPID. STUDY DESIGN Prospective observational study. METHODS Plasma ACTH concentration in 8 non-PPID horses and 8 horses with PPID was measured at 08.00, 11.00, 14.00 and 17.00 h on 5 nonconsecutive days within a 3 week period. In addition, at 08.30 h on one day, 6 samples were collected from each horse at precisely 5 min intervals over a period of 25 min. Descriptive and graphical analysis was performed and a linear mixed effects model was fitted to assess the effect of time of day on ACTH concentration in non-PPID and PPID horses. RESULTS Evidence of ultradian fluctuation in ACTH production was not identified in either non-PPID or PPID horses. Evidence for circadian fluctuation was identified in non-PPID horses; plasma ACTH concentrations were highest at 08.00 h and decreased through the day. There was no evidence of circadian fluctuation in PPID horses. In non-PPID horses, the magnitude of circadian changes in ACTH concentration was smaller than variations in concentration that occurred at random. Intrahorse variability of ACTH concentration was greater in PPID horses than in non-PPID horses. CONCLUSIONS Ultradian and circadian fluctuations in ACTH concentration are unlikely to influence clinical decision making; however, variations of potential clinical relevance do occur in individual horses, for reasons that remain to be determined, and increase in magnitude with progression of PPID. Results of the current study indicate that when an ACTH concentration between 19 and 40 pg/ml is measured, further testing should be considered to increase the accuracy of PPID diagnosis.

The efficacy of ivermectin, pyrantel and fenbendazole against Parascaris equorum infection in foals on farms in Australia

This study was performed to estimate the prevalence of patent Parascaris equorum infections and determine the efficacy of ivermectin, pyrantel and fenbendazole against P. equorum infection in foals on farms in southern Australia. Foals aged >3 months on five farms in the south-western slopes region of New South Wales were used. Faeces were collected from each foal and foals with a P. equorum faecal egg count (FEC) of >100 eggs per gram (EPG) were used to measure anthelmintic efficacy using the FEC reduction (FECR) test, after random allocation to a control group or an ivermectin, pyrantel embonate or fenbendazole treatment group. Treatment was administered on day 0 and faeces were collected on day 14 and a FEC was performed. For determination of anthelmintic efficacy, FECRs and lower 95% confidence intervals (LCL) were calculated using previously described methods, based on individual or group FECRs. P. equorum populations were considered susceptible when FECR was >90% and LCL >90%, suspected resistant when FECR was FECR was 80-90% and LCL <90% and resistant when FECR was <80% and LCL <90%. A Poisson distribution quality control method was applied to the data to remove suspected erroneous FECR results. Prevalence of patent P. equorum infection was 58.3% (147/252 foals) and 89 foals on 5 farms were included in the FECR study. Resistance of P. equorum to ≥ 1 anthelmintic was present on all five farms prior to and on four farms after application of the quality control method. Two farms had evidence of multiple drug resistance. Ivermectin was effective and ineffective on two and three farms, respectively. Fenbendazole was effective on two farms, equivocal on one farm and ineffective on one farm. Pyrantel embonate was effective on three farms and ineffective on one farm. These data indicate that anthelmintic-resistant P. equorum populations are present on farms in Australia and multiple drug resistance may occur on individual farms.

Comparative study of doramectin and fipronil in the treatment of equine chorioptic mange.

In equids, chorioptic mange is a common dermatitis for which there are no licensed medications in the uk. Doramectin and fipronil are licensed for the control of ectoparasites in other species and were evaluated for the treatment of 17 cases of chorioptic mange in 13 equids. Equids were included if clinical findings were indicative of chorioptic mange, chorioptes mites were positively identified and concurrent disease that could affect response to disease was not evident. A random number table was used to allocate subjects to receive doramectin (group D, eight animals) or fipronil (group F, nine animals). Each of the horses in group D were given 0·3 mg/kg doramectin (Dectomax; Pfizer) on two occasions 14 days apart by subcutaneous injection. All limbs of the horses in group F were sprayed with fipronil 0·25 per cent solution (Frontline; Merial) to the level of the stifles and elbows. Both groups were examined on the day of treatment and 14 and 28 days later to assess the behavioural signs of pruritus and the severity of the dermatological lesions. Acetate tape impressions were collected from the distal limbs and the degree of mite infestation was assessed. By day 28 there were no behavioural signs of chorioptic mange in any of the animals, and there were significant reductions in the numbers of mites in both groups. However, there were no significant reductions in the mean lesion score in either group. There was no significant difference between the effectiveness of the two treatments.

The application of exhaled breath gas and exhaled breath condensate analysis in the investigation of the lower respiratory tract in veterinary medicine: A review.

The analysis of biomarkers in exhaled breath (EB) and exhaled breath condensate (EBC) may allow non-invasive and repeatable assessment of respiratory health and disease in mammals. Compared to human medicine, however, research data from EB and EBC analysis in veterinary medicine are limited and more patient variables influencing concentrations of EB/EBC analytes may be present. In addition, variations in methodologies between studies may influence results. A comparison of the approaches used in veterinary research by different groups may aid in the identification of potentially reliable and repeatable biomarkers suitable for further investigation. To date, changes in acid-base status and increased concentrations of inflammatory mediators have been the main findings in studies of pulmonary disease states in animals. Whilst these biomarkers are unlikely to represent specific and sensitive diagnostic parameters, they do have potential application in monitoring disease progression and treatment response.

Assessing techniques for disinfecting sites for inserting intravenous catheters into the jugular veins of horses

The sites of insertion of catheters into the jugular veins of six horses were investigated to determine common isolates and to assess the effectiveness of two disinfection protocols with the hair coat left long, clipped or shaved. Skin commensals (Staphylococcus, Streptococcus and Micrococcus species) and environmental contaminants (Bacillus, Enterobacteriaceae, Aspergillus and Mucor species) were the microorganisms most frequently isolated. Chlorhexidine gluconate and povidone-iodine-based skin disinfection protocols resulted in significant reductions in the number of bacterial isolates from clipped sites. With chlorhexidine, there were no significant differences between the reductions observed at sites with the hair coat left long, clipped or shaved.

Stability of common biochemistry analytes in equine blood stored at room temperature

REASONS FOR PERFORMING STUDY Time delays between collection of blood samples and biochemical analysis of equine blood are unavoidably common in equine practice. The effect that delays may have on the accuracy of results of blood biochemical analyses is not well established. HYPOTHESIS Delays in processing of blood of up to 72 h results in alterations in measured levels of common biochemical analytes that are of potential clinical relevance. Separation of serum prior to storage is protective against the effects of time delays. METHODS Samples of clotted blood, separated serum and oxalate fluoride plasma from 20 horses were stored and analysed at 0, 24, 48 and 72 h. Graphical exploration of each analyte was undertaken. General linear models with fixed effects were fitted for the whole blood data. The mean bias and 95% limits of agreement were calculated, using bootstrapped data, to assess agreement between pairs of samples analysed at 0 h and other time points. Bland-Altman plots were used to explore general trends in the data. Paired t tests were used to compare the results from whole blood and separated serum. RESULTS Delays in processing equine blood resulted in significant increases in measured concentrations of aspartate aminotransferase, creatine kinase, lactate dehydrogenase, total bile acids and magnesium. A significant decrease in concentration was identified for glucose (serum and oxalate fluoride preserved plasma). Separation of serum immediately following clot formation resulted in nonsignificant increases in accuracy for some analytes. CONCLUSIONS AND PRACTICAL SIGNIFICANCE: Delays in processing of blood samples may result in biochemical changes of clinical relevance in individual cases; however, in the majority of cases, where delays are only a few days and a number of analytes are assessed concurrently, delays are unlikely to have an effect on the interpretation of results. Separation of serum following clot formation is of limited benefit. Clinical samples in which a delay in processing has occurred may be interpreted with reference to the data presented.

Multicentric T-cell lymphoma presenting as inferior palpebral swelling in a Standardbred mare

A 6-year-old Standardbred mare that presented with bilateral palpebral swelling and a left corneal ulcer was diagnosed with a multicentric T-cell lymphoma. Other clinical findings included submandibular lymphadenopathy and oedema, pharyngeal oedema and rhinitis. Prior to euthanasia the horse developed keratomalacia, the onset of which was coincidental with the use of high doses of topical and systemic corticosteroids. Although an unusual presentation, palpebral swelling should be considered as a clinical sign of lymphoma.