R. J. Naylor

Comparison of flunixin meglumine and meloxicam for post operative management of horses with strangulating small intestinal lesions

REASONS FOR PERFORMING STUDY Ex vivo evidence suggests that cyclo-oxygenase (COX) 2-preferential inhibitor nonsteroidal anti-inflammatory drugs (NSAIDs), such as meloxicam, have a less detrimental effect on intestinal healing than flunixin meglumine (FM). Whether this translates to a beneficial effect in horses with naturally occurring strangulating small intestinal (SSI) lesions is unknown. OBJECTIVES To compare the clinical outcome of horses with naturally occurring SSI lesions treated with meloxicam or FM. STUDY DESIGN Randomised prospective study. METHODS Cases presenting to the Royal Veterinary College Equine Referral Hospital and Bell Equine Veterinary Clinic during 2010 and 2011 in which an SSI lesion was identified at exploratory laparotomy were eligible for inclusion. Horses received either 1.1 mg/kg bwt FM or 0.6 mg/kg bwt meloxicam i.v. q. 12 h. Clinical outcomes and clinical and laboratory parameters associated with endotoxaemia were compared between groups. RESULTS Sixty cases were enrolled, 32 horses received FM and 28 received meloxicam. There was no difference in signalment, physical examination or surgical factors between groups. The overall survival to discharge was 81%; there was no difference in survival (P = 0.14) or incidence of post operative ileus (P = 0.25) between groups. There was no significant difference between the plasma lipopolysaccharide (LPS) concentrations at 0 h (P = 0.18) or 48 h (P = 0.60); however, there was a significant difference between neutrophil count at 48 h (P<0.05) and at 96 h (P<0.01) with significantly greater cell numbers in horses receiving meloxicam compared with FM. Blinded pain score evaluation showed that more horses receiving meloxicam showed gross signs of pain than those treated with FM (P = 0.04). CONCLUSIONS Nonsteroidal anti-inflammatory drug choice did not affect major clinical outcomes in horses with SSI lesions but had some effects on signs of pain. This study provides no evidence to recommend one NSAID treatment above another based on survival or the incidence of ileus; however, evaluation of a larger number of cases is required.

Histopathology and computed tomography of age‐associated degeneration of the equine temporohyoid joint

REASONS FOR PERFORMING STUDY The aetiology of temporohyoid osteoarthropathy (THO) is unknown; both primary infectious and degenerative causes have been suggested. HYPOTHESIS There is a significant association between increasing age and severity of temporohyoid joint degeneration. To examine the histopathology of the temporohyoid articulation in aged horses and to compare the appearance of the joint with computed tomography (CT) and peripheral quantitative CT (pQCT). METHODS pQCT scans of the temporohyoid articulations were obtained bilaterally from 31 horses (range age 1-44 years) post mortem and images were graded by 2 blinded observers on 2 occasions for the presence of osteophytes, irregularity of the joint surface and mineralisation. Eight heads had been examined previously by CT, with the images similarly graded for the shape and density of the proximal stylohyoid bones, bone proliferation surrounding the joint, mineralisation of the tympanohyoid cartilage and the relationship of the petrous temporal bone to the stylohyoid bone. Sixteen temporohyoid joints were then evaluated histologically. RESULTS There was significant association between the mean pQCT degeneration score and age (rho = 0.75; P<0.0001), between the pQCT and CT score (rho = 0.63; P = 0.01) and between the degenerative changes identified within each temporohyoid joint within each horse (rho = 0.81; P<0.0001). Age-associated changes included the development of a club shape by the proximal stylohyoid bone, rounding of the synostosis with the petrous temporal bone and extension of osteophytes from the petrous temporal bone to envelope the stylohyoid head and bridge the joint. In no horse was there any evidence of osteomyelitis within the petrous temporal bone, stylohyoid bone or tympanohyoid cartilage. CONCLUSIONS This study provides evidence that age is associated with increasing severity of degenerative changes in the equine temporohyoid joint and that similar changes are commonly found bilaterally. POTENTIAL RELEVANCE The changes identified appear similar, albeit milder to the changes reported in horses with THO, suggesting that degenerative, rather than infectious causes may underlie the aetiology of THO. Future work should be directed at examining the histopathology of clinical THO cases.

Allele Copy Number and Underlying Pathology Are Associated with Subclinical Severity in Equine Type 1 Polysaccharide Storage Myopathy (PSSM1)

Equine type 1 polysaccharide storage myopathy (PSSM1), a common glycogenosis associated with an R309H founder mutation in the glycogen synthase 1 gene (GYS1), shares pathological features with several human myopathies. In common with related human disorders, the pathogenesis remains unclear in particular, the marked phenotypic variability between affected animals. Given that affected animals accumulate glycogen and alpha-crystalline polysaccharide within their muscles, it is possible that physical disruption associated with the presence of this material could exacerbate the phenotype. The aim of this study was to compare the histopathological changes in horses with PSSM1, and specifically, to investigate the hypothesis that the severity of underlying pathology, (e.g. vacuolation and inclusion formation) would (1) be higher in homozygotes than heterozygotes and (2) correlate with clinical severity. Resting and post-exercise plasma creatine kinase (CK) and aspartate aminotransferase (AST) enzyme activity measurements and muscle pathology were assessed in matched cohorts of PSSM1 homozygotes, heterozygotes or control horses. Median (interquartile range (IR)) resting CK activities were 364 (332–764) U/L for homozygotes, 301 (222–377) U/L for heterozygotes and 260 (216–320) U/L for controls, and mean (+/− SD) AST activity for homozygotes were 502 (+/116) U/L, for heterozygotes, 357 (+/−92) U/L and for controls, 311 (+/−64) U/L and were significantly different between groups (P = 0.04 and P = 0.01 respectively). Resting plasma AST activity was significantly associated with the severity of subsarcolemmal vacuolation (rho = 0.816; P = 0.01) and cytoplasmic inclusions (rho = 0.766; P = 0.01). There were fewer type 2× and more type 2a muscle fibres in PSSM1-affected horses. Our results indicate that PSSM1 has incomplete dominance. Furthermore, the association between plasma muscle enzyme activity and severity of underlying pathology suggests that physical disruption of myofibres may contribute to the myopathic phenotype. This work provides insight into PSSM1 pathogenesis and has implications for related human glycogenoses.

Blood lactate concentrations in ponies and miniature horses with gastrointestinal disease.

REASONS FOR PERFORMING STUDY Clinical impression suggested that pony and miniature breeds (collectively referred to as ponies) presenting to a referral hospital for investigation of gastrointestinal disease had higher blood lactate concentrations on admission than large breed horses. OBJECTIVES The study tested the hypothesis that ponies with gastrointestinal disease had higher blood lactate concentrations on admission than large breed horses with similar disease severity. STUDY DESIGN Retrospective case-control study. METHODS Medical records from September 2006 to July 2011 were reviewed for ponies with a primary presenting complaint of gastrointestinal disease. Two larger breed horses with gastrointestinal disease were selected as controls for each case. Data collected included case details, historical and clinicopathological findings, diagnosis and outcome. RESULTS Information was collected on 50 ponies and 100 horses. Ponies had higher mean ± s.d. respiratory rates (27 ± 13 vs. 21 ± 13 beats/min; P = 0.01) and rectal temperatures (37.9 ± 0.6 vs. 37.4 ± 0.6°C; P = 0.006) and a longer median duration of clinical signs prior to presentation (10 h [1-72 h] vs. 6 h [1-120]; P<0.001). Median blood lactate concentrations on admission were higher in ponies than in horses (2.8 mmol/l [0.7-18.0] vs. 1.6 mmol/l [0.4-8.1]; P = 0.001). All other parameters relating to colic severity were not significantly different between groups, although more horses underwent exploratory laparotomy (19/50 ponies and 55/100 horses; P = 0.05). Median blood lactate concentrations in ponies with large intestinal disease, nonstrangulating lesions, undergoing medical treatment and surviving ponies were significantly higher than in horses in the same category. In contrast to horses, no differences in blood lactate concentrations exist between ponies with medical vs. surgical treatment, strangulating and nonstrangulating lesions and surviving and nonsurviving ponies. CONCLUSION AND POTENTIAL RELEVANCE Ponies might present with higher blood lactate concentrations than horses and might falsely be suspected of having a surgical lesion or a poorer prognosis if veterinarians are not aware of breed differences.

Evaluation of Cardiac Phenotype in Horses with Type 1 Polysaccharide Storage Myopathy

BACKGROUND Type 1 polysaccharide storage myopathy (PSSM1), an equine glycogen storage disorder caused by a gain of function mutation (R309H) in the gene encoding glycogen synthase (GYS1), is associated with the accumulation of amylase-resistant alpha-crystalline polysaccharide inclusions within skeletal muscle. Several glycogenoses in humans have a cardiac phenotype, and reports exist of horses with PSSM and polysaccharide inclusions in cardiac muscle. HYPOTHESIS/OBJECTIVES To investigate the hypothesis that horses with PSSM1 display a cardiac phenotype. Our objectives were to compare plasma cardiac troponin I (cTnI) concentration and the incidence of cardiac arrhythmias in PSSM1 homozygotes, heterozygotes, and control horses. METHODS One hundred and twenty-five Belgian and Percheron horses under the same management were genotyped for the R309H GYS1 mutation. From these, 8 age-, breed-, and sex-matched cohorts of each genotype were identified. Plasma cTnI concentration and incidence of cardiac arrhythmias (determined by 24-hour Holter ECG) were compared between the groups. RESULTS Although some PSSM1-affected horses had mildly increased plasma cTnI concentrations, there was no significant difference in cTnI concentrations between groups. There were no significant differences in the incidence of ectopic beats, cardiac conduction intervals or mean heart rate between groups. CONCLUSIONS AND CLINICAL IMPORTANCE We found no evidence of clinically relevant cardiac myocyte injury or arrhythmias in horses with PSSM1. Additional study is required to determine whether myocardial function may be compromised in this disorder.

Suspected acorn toxicity in nine horses

REASONS FOR PERFORMING STUDY Acorn toxicity has been anecdotally reported to cause fatal colitis and colic in horses but reports in the scientific literature are sparse. OBJECTIVES This study reports the diagnosis, treatment, prognosis and outcome of 9 cases with suspected acorn toxicity admitted to 2 referral hospitals. STUDY DESIGN Retrospective case series. METHODS Case records from 2004 to 2013 were reviewed. Horses were included in the study if they met 3 of 4 criteria: exposure to acorns; clinical and laboratory data suggesting alimentary or renal dysfunction; acorn husks in the faeces or gastrointestinal tract; and necropsy and histopathological findings consistent with acorn toxicity. Data collected included case history, clinical presentation, clinicopathological data, ultrasonographic findings, case progression, and necropsy and histopathological findings. RESULTS Nine horses met the inclusion criteria. Five cases presenting with haemorrhagic diarrhoea deteriorated rapidly and were subjected to euthanasia or died. Four cases showed signs of colic with gas distension, displacement of the large colon and diarrhoea. Three of these (33%) survived with medical management, the fourth was subjected to euthanasia. Post mortem examination of 6 cases demonstrated submucosal oedema of the large intestine and caecum (n = 6), acute tubular nephrosis (n = 6), diffuse necrohaemorrhagic and ulcerative typhlocolitis and enteritis (n = 4), and small intestinal oedema (n = 3). CONCLUSIONS Acorn ingestion may be associated with typhylocolitis leading to diarrhoea, colic and acute renal tubular nephrosis. Recovery is possible in mildly affected cases; more severe cases show hypovolaemia, intractable pain, renal dysfunction and cardiovascular failure, and often succumb to the disease process. Disease is only seen in a small proportion of the population exposed to acorns and there seems to be an increased occurrence in certain years. Further investigation into factors predisposing to disease is required, but limiting exposure to acorns in the autumn seems prudent.

The treatment of diarrhoea in the adult horse

Summary The priority in treating the equine patient with acute diarrhoea is to stabilise the haemodynamic aberrations secondary to the fluid and electrolyte losses. Once this has been initiated and the patient is stabilised ancillary treatments may be beneficial. Besides the well established effects of NSAIDs and polymixin B on systemic inflammation, recent studies suggest that the use of DTOS to bind bacterial toxins and Saccharomyces boulardii to reduce the severity and duration of diarrhoea may be beneficial. The justification for using probiotic products is scant. There is no evidence to suggest that systemic use of antimicrobials benefits equine patients with colitis, with the exception of metronidazole in cases of clostridial diarrhoea. In light of their potentially detrimental effects, their use can, in the opinion of the authors, not be advocated. Better understanding of the pathways of systemic inflammation and more selective anti-inflammatory drugs may be of great benefit in the future.

Standing CT and clinical progression of equine cholesterol granulomata.

CHOLESTEROL granulomata – benign choroid plexus masses often detected at equine postmortem examinations – are sometimes associated with central neurological signs (Ivoghli and others 1977, Johnson and others 1993, Jackson and others 1994, King 1997). Antemortem diagnosis was impossible until advent of magnetic resonance imaging (Maulet and others 2008) and CT (Vink-Nooteboom and others 1998, Vanschandevijl and others 2008). These two cases are the first to illustrate diagnosis using standing (unanaesthetised) CT (CT Lightspeed QX/i; GE medical systems) and describe the long-term follow-up of one case, revealing that the prognosis for some cases is fair. Case 1: A 12-year-old Irish Draught cross gelding was presented with a 10-day history of intermittent lethargy and abnormal head carriage. On presentation, episodes of generalised muscle rigidity and ataxia were noticed by the owner; the horse was obtunded with reduced tongue tone and drooping of the lower lip and reluctant to flex or extend the neck and back. Neurological signs had improved following administration of methylprednisolone, penicillin and flunixin meglumine. On presentation to the Royal Veterinary College, the horse was obtunded; menace responses were reduced bilaterally, but pupillary light reflexes were intact. Mild superficial gluteal muscle fasciculations were evident. The horse had an exaggerated guarding response to deep palpation of the neck at C5 to C7. The neuroanatomical localisation suggested forebrain involvement, but based on cervical discomfort on …

Development of a clonal equine myoblast cell line capable of terminal differentiation into mature myotubes in vitro

OBJECTIVE To produce a clonal equine myoblast cell line that retains the ability to divide for multiple passages and differentiate into multinucleated myotubes during specific conditions. SAMPLE Cultured primary equine skeletal muscle-derived cells from a healthy Thoroughbred. PROCEDURES Cell cultures were transfected by electroporation with a plasmid (pNIT) that expresses the temperature-sensitive simian vacuolating virus 40 large T antigen (TAg), which can be controlled by a doxycycline-responsive promoter. Cells that stably integrated the TAg were selected and expanded to passage 25. For each passage, differentiation and fusion properties of the cells were determined and immunocytochemical analyses were performed to evaluate expression of TAg and other muscle-specific proteins. Optimum conditions that led to cell differentiation into myotubes were also determined. RESULTS Compared with nontransfected control cells, myogenic, desmin-positive cells expressed the TAg when incubated at 33°C and could be maintained in culture for numerous passages. Reduced expression of TAg was identified in cells incubated at 37°C or when incubated with doxycycline at 33°C. Expression of TAg was not detected when cells were incubated with doxycycline at 37°C, and when serum was withdrawn from the culture medium, those clones differentiated into a pure population of multinucleated myotubes. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that production of an immortalized clonal equine skeletal muscle cell line was possible. A clonal equine skeletal muscle cell line will be a valuable in vitro tool for use in equine physiology and disease research.

Severe hyperkalaemia associated with renal dysplasia in a 2-day-old foal

Summary A 2-day-old filly foal presented with signs of depression, recumbency and inappetence. Blood analyses revealed hypoalbuminaemia, hyperfibrinogenaemia, hyperglycaemia and hyperkalaemia. The foal deteriorated despite intensive treatment and was subjected to euthanasia. At post mortem examination, the urinary bladder, ureters and kidneys appeared normal grossly. Histologically both kidneys showed disorganised development with the presence of structures inappropriate for a foal of this age, including primitive glomeruli, immature renal tubules and persistent metanephric ducts. Based on these findings a diagnosis of bilateral renal dysplasia was made.