Identification and kinetics of microsomal and recombinant equine liver cytochrome P450 enzymes responsible for in vitro metabolism of omeprazole.

In humans, omeprazole is metabolised by cytochrome P450 (CYP450) CYP2C19 and CYP3A4 with differences in CYP2C19 genotypes leading to variable response to therapy. Despite a wide use of omeprazole in horses with evidence of variable therapeutic efficiency, information regarding enzymatic metabolism is not currently available. This study aims to describe the in vitro kinetics of omeprazole metabolism and determine which enzyme(s) are responsible for omeprazole metabolism in horses. Omeprazole (0-800 uM) was incubated with liver microsomes and a panel of equine recombinant CYP450s (eq-rCYP). Metabolite concentrations were quantified by LC-MS and the kinetics of metabolites' formation were calculated by non-linear regression analysis. The in vitro liver microsomes formed three metabolites (5-hydroxy-omeprazole, 5-O-desmethyl-omeprazole and omeprazole-sulfone). The 5-O-desmesthyl-omeprazole formation was best fitted to a two enzyme Michaelis-Menten (MM) model with the high affinity site Clint double that of the low affinity site. For 5-hydroxy-omeprazole the best fit was to a 1 enzyme MM model with a Clint higher than for 5-O-desmesthyl-omeprazole (0.12 vs 0.09 pmol/min/pmol P450). The formation of omeprazole-sulfone was negligible. Recombinant CYP3A89 and CYP3A97 produced substantial amounts of 5-hydroxy-omeprazole (1551.72 ng/mL and 1665.33 ng/mL, respectively), while 5-O-desmethyl-omeprazole and omeprazole-sulfone were formed to a much lesser extent by multiple eq-rCYP from the CYP2C and CYP3A family. In vitro metabolism of omeprazole in horses is different to that in humans, with major metabolites produced by the CYP3A family. The current study provides the basis for further investigations of CYP450 single nucleotide polymorphisms that could affect omeprazole metabolism and therapeutic efficacy.

Horse Grimace Scale Does Not Detect Pain in Horses with Equine Gastric Ulcer Syndrome.

Equine gastric ulcer syndrome (EGUS) is a highly prevalent and presumptively painful condition, although the amount of pain horses might experience is currently unknown. The aims of this study were to determine if the Horse Grimace Scale (HGS) could identify pain behaviours in horses with and without EGUS and if severity would be positively associated with the HGS score. Horse grimace scale scores were assessed blindly using facial photographs by seven observers and involved evaluation of 6 facial action units as 0 (not present), 1 (moderately present) and 2 (obviously present). Lameness examination, serum amyloid A (SAA) measurement and gastroscopy evaluation were performed on all horses. Horses (n = 61) were divided into two and three groups based on the presence (yes, no) and severity (none, mild, moderate-severe) of EGUS, respectively. Presence of lameness and elevated SAA (≥50 µg/mL) were used as exclusion criteria. Inter-observer reliability was analyzed by intra-class correlation coefficients (ICC). HGS scores between groups were compared using Welch's and Brown Forsythe tests (p < 0.05). Overall, HGS ICC was "excellent" (0.75). No significant differences (p = 0.566) were observed in HGS scores between horses with and without gastric lesions (mean, 95% CI; 3.36, 2.76-3.95 and 3, 1.79-4.20, respectively). HGS was not influenced by the presence or severity of EGUS in this current study. Further studies investigating the use of different pain scales in horses with EGUS are needed.

Eosinophilic Inflammation and Equine Herpesvirus-1 Associated With Haemorrhagic Cystitis in a Horse. Case Report.

Equine idiopathic haemorrhagic cystitis (EIHC) is a recently described form of aseptic cystitis in horses in which there is no discernible underlying cause. This case report describes a 9-year-old Thoroughbred gelding that presented with stranguria, pollakiuria, and haematuria. Cystoscopy revealed ulceration and haemorrhage of the bladder mucosa, diffuse mural hyperaemia and marked urine sedimentation. Histopathological evaluation of the bladder revealed chronic active ulcerative neutrophilic, lymphoplasmacytic, and eosinophilic cystitis. There was no bacterial or fungal growth upon culture but polymerase chain reaction (PCR) testing and sequencing for equine herpesvirus-1 (EHV-1) on bladder mucosa was positive. Conservative therapy with broad spectrum antimicrobials and non-steroidal anti-inflammatory therapy yielded complete resolution of clinical signs with significant improvement of macroscopic lesions in 14 days. Although a positive EHV-1 PCR suggests a viral cause, the horse's clinical signs, histology and recovery rate are more consistent with equine idiopathic haemorrhagic cystitis (EIHC). Neutrophilic and lymphoplasmacytic inflammation is a known feature of EIHC but eosinophilic infiltrates have not been previously described. The significance of the eosinophilic involvement is not certain; however, their presence has been associated with fungal, viral, parasitic, and immune-mediated aetiologies in other body systems. This is the first report of a horse with possible EIHC in Australia, as well as the first case with eosinophilic infiltrates and testing positive for EHV-1.

Bilateral thyroid adenomas in an alpaca.

A 7-year-old neutered male alpaca (Vicugna pacos) was presented for evaluation of a 3-year history of large, bilateral, firm ventral cervical masses causing esophageal and tracheal impingement. Ultrasound examination, radiographic evaluation, histopathological findings, and magnetic resonance imaging confirmed the masses to be bilateral thyroid adenomas. Conservative medical treatment by unilateral chemical ablation, using 10% formalin by aspiration technique, was performed on the left mass. Chemical ablation proved to be effective in decreasing the size of the mass, with no apparent adverse effects. To our knowledge, this case is the first known report of bilateral thyroid adenomas in an alpaca, a condition previously described in humans, horses, dogs, and cats.