Gabriele Rossi

Background
Lipoprotein fractions are reported to be unstable in stored human samples, and there is a paucity of information on the analytical precision of electrophoretic separation of lipoproteins in canine serum samples.

Objective
The aim of this study was to assess the effects of intra- and inter-assay imprecision and of storage conditions on the electrophoretic separation of canine lipoproteins.

Methods
Imprecision was assessed by calculating the coefficient of variation (CV) of five replicates of six serum samples run in two sequential runs of agarose gel lipoprotein electrophoresis. The effect of storage was assessed with a Friedmann test by comparing the results of samples analyzed after sampling (T0) and after 24 and 48 h at room temperature or stored at 4°C and after 7, 14, 21 days, 1, 2, and 3 months at −20°C or at −80°C. Moreover, electrophoretograms obtained after storage were visually analyzed by two observers in a blind manner to assess whether storage alters the electrophoretic profile.

Results
The imprecision of high-density lipoproteins (HDL), low-density lipoproteins (LDL), very low-density lipoprotein (VLDL), and chylomicrons were respectively 0.8%–11.5%, 2.4%–22.7%, 2.3%–11.5%, and 12.5%–105.2%. Compared with T0, HDL significantly decreased, and LDL significantly increased over time in all the storage conditions, whereas VLDL significantly increased only in frozen samples, and chylomicrons did not significantly differ. In frozen samples, deviations from baseline values were lower than the imprecision of the method, and visual misclassifications of electrophoretograms were rare.

Conclusions
Despite minimal variation in the percentage of some fractions, freezing does not influence the interpretation of canine lipidograms.

Objective:
To explore changes in urinary biomarkers of acute kidney injury (AKI) in healthy dogs experiencing intra-operative hypotension and explore the relationship between blood pressure and urinary biomarkers.

Study Design:
Observational cohort study.

Animals:
A group of 50 client owned dogs.

Methods:
Urine and blood samples were collected prior to anaesthesia (T0), within 24 hours after anaesthesia (T1) and 10 days post-surgery (T10). During anaesthesia, the lowest mean arterial pressure (MAP) in each dog. Impact of duration was explored by categorising according to arbitrary thresholds of MAP < 50, < 60, < 70 and < 80 mmHg and calculating duration (minutes) within each category Serum creatinine (Cr) and validated biomarkers of AKI including urinary gamma-glutamyl transferase (uGGT), urinary neutrophil gelatinase-associated lipocalin (uNGAL), and urinary cystatin C (uCystatin C) were measured. Biomarker measurements were standardised to urinary Cr. The frequency of dogs with proportional increases between T1 and T0 and between T10 and T0 was recorded. Multiple regression analysis was performed to determine the simplest subset of independent variables (lowest MAP, duration with each MAP category) to best explain the variance in the proportional change of each biomarker.

Results:
Hypotension, defined as MAP < 60 mmHg was observed in 38/50 (76 %) of the dogs. Between T1 and T0, increases in uGGT/Cr, uCystatin C/Cr, and uNGAL/Cr were observed in 37 (82%), 17 (41 %) and 19 (35%) of 50 dogs, respectively. Of the variance observed in uGGT/Cr at T1/T0, 62% could be explained by the lowest MAP recorded when combined with duration MAP < 50mmHg (adjusted R2 0.62).

Conclusion and clinical relevance:
In this clinical model of intra-operative hypotension uGGT/Cr demonstrated potential for diagnosis of early AKI in healthy dogs. Increases in validated biomarkers uCystatin C/Cr, and uNGAL/Cr support their use in future studies investigating different causes or severity of AKI.

Through evaluation of serum and plasma buterylcholinesterase (BChE) and brain acetylcholinesterase (AChE) activity, we investigated the possibility of the involvement of an acute organophosphate toxicosis in the pathogenesis of ongoing annual outbreaks of paresis and paralysis that in some cases progress to death, in endangered Western Australian Carnaby's cockatoos (Zanda latirostris). The condition, named Carnaby's hindlimb paralysis syndrome (CHiPS), was first described in 2012. Following initial investigations involving clinical, epidemiologic, toxicologic, gross necropsy, and histologic evaluation, a toxic etiology, specifically an organophosphate toxicosis, was considered most likely. The study aimed to validate the BChE assay for use in serum and plasma in Carnaby's cockatoos. This study found no evidence of changes in serum or plasma BChE or brain AChE that indicate an acute organophosphate toxicosis as the cause of CHiPS. Although these results render an acute organophosphate toxicosis unlikely, an organophosphate-induced delayed neuropathy has not been ruled out. Based on the results from the BChE validation study, the authors can recommend this assay for the evaluation of BChE measurement in plasma and serum from Carnaby's cockatoos with results showing excellent accuracy and precision.

Introduction: Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are prescribed to manage hyperinsulinemia but the effects of dapagliflozin have not been investigated. Although hyperlipaemia is very rare, hypertriglyceridemia is commonly associated with SGLT2i treatment and investigation of the lipoprotein profiles is warranted.

Methods: Retrospective analysis of clinical records and stored serum from horses with hyperinsulinemia that received dapagliflozin (0.02 mg/kg, [n = 34]) or ertugliflozin (0.05 mg/kg [n = 24]) PO SID for 30 days. Within-horse changes, correlations between variables, and differences between treatments were assessed using Wilcoxon signed-rank, Spearman's rank correlation coefficient (rho) and the medians tests, respectively.

Results: Between day 0 (pre-treatment) and day 30 within-horse changes (median, inter-quartile range [IQR]) were: basal serum [Insulin] (uU/mL) reduced from 170 (92-280) to 28.7 (14.5-90) (P < .0001), lameness grade (scale 0-12) reduced from 6 (4-10) to 2 (0-2) (P < .001), serum [triglyceride] (mmol/L) increased from 0.5 (0.3-0.6) to 1.0 (0.6-1.56), [β-hydroxybutyrate] (μmol/L) increased from 0.22 (0.17-2.7) to 0.30 (0.24-0.35) (P < .0001), [total cholesterol] (mmol/l) increased from 2.36 (2-2.6) to 2.84 (2.4-3.7) (P < .0001) and, as a percentage of serum lipids, high-density lipoprotein (HDL) reduced from 52.4% (47.9%-61.0%) to 50% (41%-54.8%) (P = .034), very-low density lipoprotein (VLDL) increased from 10.4% (6.4%-14.4%) to 12.3% (9.9%-16.8%) (P = .005). Differences between ertugliflozin and dapagliflozin groups in these parameters were not significant at day 0 or 30. At day 30, 10/48 (21%) cases had [triglycerides] >2.0 mmol/L (maximum = 10.8 mmol/L). Day 30 [triglyceride] was correlated with day 0: basal insulin (P < .001, rho = 0.47), [triglyceride] (P = .003, rho = 0.42) and %VLDL (P = .019, rho = 0.34) and day 30: [total cholesterol] (P < .001, rho = 0.67), %HDL (rho = −0.432, P = .014) and %VLDL (rho = 0.708, P < .001).

Discussion and clinical relevance: Dapagliflozin or ertugliflozin treatment is associated with reductions in [insulin] and lameness grade. Changes in [triglyceride] and lipoprotein profiles were usually minor with occasional marked hypertriglyceridemia. [β-hydroxybutyrate] increased indicating ketosis, a metabolic pathway previously not thought to be relevant in horses.

ORAL PRESENTATION ACVIM Resident Research Abstract Award Winner

The metabolic and lipid profiles of horses treated with sodium-glucose cotransporter 2 inhibitors are not well understood. This retrospective study evaluated blood parameters in hyperinsulinemic horses treated with either ertugliflozin (0.05 mg/kg) or dapagliflozin (0.02 mg/kg) orally once daily. Blood samples were collected at baseline (day 0) and after 7 and/or 30 days of treatment. Statistical analyses were conducted using Wilcoxon signed-rank, Mann-Whitney and Spearman's rank correlation tests. Thirty-four horses received dapagliflozin and 24 received ertugliflozin. Significant (p<0.05) within-horse changes between day 0 and day 30 included [median, inter-quartile range (IQR)]: basal serum [Insulin] (uU/ml) reduced 170 (92-280) to 28.7 (14.5-90); [triglycerides] (mmol/l) increased 0.5 (0.3-0.6) to 1.0 (0.6-1.56), [β-hydroxybutyrate] (umol/l) increased 0.22 (0.17-2.7) to 0.30 (0.24-0.35); [total cholesterol] (mmol/l) increased 2.36 (2-2.6) to 2.84 (2.4-3.7); and GGT (IU/ml) increased 21 (16-32) to 25 (18-38). As a percentage of total serum lipids, high-density lipoprotein (HDL) reduced 52.4 % (47.9 %-61.0 %) to 50 % (41 %-54.8 %) and very-low density lipoprotein (VLDL) increased 10.4 % (6.4 %-14.4 %) to 12.3 % (9.9 %-16.8 %) (all p<0.05). Differences between ertugliflozin and dapagliflozin groups were not significant in any of these parameters at days 0, 7 or 30. At day 30, 10/48 (21 %) cases had [triglycerides] > 2.0 mmol/l (maximum = 10.8mmol/l). Day 30 [triglyceride] correlated with day 0: basal insulin (rho=0.47); [triglyceride] (rho=0.42); %VLDL (rho=0.34) day 30: [total cholesterol] (rho=0.67), %HDL (rho=-0.432) and %VLDL (rho=0.708). Our findings suggest that SGLT2 inhibitors induce minor changes in lipid profiles, with occasional cases of marked hypertriglyceridemia, and that dapagliflozin and ertugliflozin exhibit similar biochemical effects.

Background
Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are being used increasingly in equine practice. While there is emerging clinical evidence of the safety and efficacy of these drugs, there are currently no reports to document owner experiences with treatment.

Objective
The objective of the study was to report owner experiences and observations following treatment with SGLT2i in horses.

Study design
A cross-sectional online survey.

Methods
Horse owners were recruited via social media, online forums and their veterinarians to participate in an anonymous online survey to document their experiences and observations when treating their horses with SGLT2i.

Results
Three hundred forty-two responses met the inclusion criteria. Ertugliflozin was the most commonly prescribed SGLT2i (79.8%), and the most common reasons for treatment were high insulin concentrations (84.2%) and active laminitis (59.7%). 85.3% of owners reported their horses had an improved quality of life after commencing treatment, while 9.4% reported no change and 5.3% reported a worsening of clinical signs. Of owners who had considered euthanasia prior to treatment (n = 77), 80.5% reported their horse's level of pain to be either mild or absent after 30 days of treatment and 94.8% reported their horse's quality of life to be improved. Most owners (n = 220, 64.7%) reported they were either extremely satisfied or somewhat satisfied (n = 72, 21.2%) with treatment. Treatment concerns included safety/side effects, medication cost, availability and long-term efficacy. 114 owners (33.3%) reported one or more initial adverse effects upon induction onto the medication, particularly excessive urination (n = 70, 20.5%), excessive drinking (n = 38, 11.1%), excessive weight loss (n = 34, 9.9%) and dullness (n = 26, 7.6%).

Main limitation
Sampling bias through social media and veterinary practices and reliance on subjective owner reports.

Conclusion
The use of SGLT2i in horses was associated with excellent rates of owner satisfaction and owner-reported improved quality of life for the horse; however, some adverse effects were observed.

Malaria and other haemosporidian parasites are common in reptiles. During baseline health surveys of sea turtles in Western Australia (WA), haemosporidian parasites were detected in flatback (Natator depressus) and green (Chelonia mydas) turtle erythrocytes during routine blood film examination. 130 blood samples were screened via polymerase chain reaction (PCR), including 105 N. depressus, 20 C. mydas, and 5 olive ridley turtles (Lepidochelys olivacea). A novel Haemocystidium sp. was identified, detected exclusively in foraging turtles and not in nesting turtles. The combined prevalence by microscopic and molecular methods was 16.9% (22/130), primarily affecting immature C. mydas (77.3%; 17/22). Mature N. depressus were also affected (22.7%; 5/22). DNA sequencing of a partial fragment of the mitochondrial cytochrome b (cytb) gene together with phylogenetic analysis identified two different Haemocystidium sp. genotypes, A and B, with genotype A being most prevalent. The phylogenetic analysis showed close genetic relationships to Haemocystidium sp. in freshwater and terrestrial turtles, suggesting a shared evolutionary lineage despite ecological differences. Preliminary analysis indicates that this parasite is incidental, as no association between health and parasite presence or grade was detected. This study provides the first formal detection of haemosporidian parasites in sea turtles, contributing essential baseline data while highlighting their evolutionary significance and host–parasite ecological relationships.

Sarcoptic mange, caused by epidermal infection with Sarcoptes scabiei, negatively impacts the health, welfare, and local abundance of bare-nosed wombats (Vombatus ursinus) in Australia. Improved understanding of the host immune response to disease and its contribution to pathophysiology could be used to inform management actions for this species in and ex situ. To evaluate the immune response of bare-nosed wombats to sarcoptic mange, we validated three assays (haptoglobin, agarose gel electrophoresis, and micro-erythrocyte sedimentation rate) measuring non-specific markers of inflammation using serum samples from free-living wombats from Tasmania (n = 33). We then analysed correlations between the assay results for each non-specific marker of inflammation and wombat’s sarcoptic mange scores, and performed histopathological examinations to investigate association of the acute phase response with systemic amyloidosis. We present evidence that haptoglobin and erythrocyte sedimentation rate increased, and albumin decreased, in association with sarcoptic mange scores. This research demonstrates links between the acute phase response and sarcoptic mange severity in bare-nosed wombats, highlighting the utility of non-specific markers of inflammation for aiding assessment of the systemic effects of mange. Showing the value of agarose gel electrophoresis, we also identified specific acute phase proteins warranting future evaluation and found evidence of an immunoglobulin response in mange-affected wombats, revealed by increasing γ-globulins in association with apparent disease severity. Meanwhile, owing to its relatively low resource requirements and rapidity, the erythrocyte sedimentation rate assay may be useful as a point-of-care test to support therapeutic decisions in the field. Our methods and findings are likely to be applicable to a range of other clinical and population health scenarios in captive and free-living wombats, and species impacted by sarcoptic mange globally.

Introduction: Dopaminergic agonists are accepted as the most effective treatment for pituitary pars intermedia dysfunction. However, some horses are refractory to daily oral pergolide, the recommended registered treatment. Extended-release cabergoline (ERC) injection may offer an alternative. The objective of this retrospective case series was to describe clinical and endocrinological responses to ERC.

Methods: Medical records of horses treated with weekly intramuscular injections of ERC (5 mg/mL, BOVA Aus) at either 0.01 mg/kg (high dose, HD) (n = 10) or 0.005 mg/kg (low dose, LD) (n = 30) were reviewed. Short-term ACTH responses were assessed at 5–8 days using a Wilcoxon signed ranked test. Longer-term ACTH responses (30 to 365 days) were assessed using generalised estimating equations.

Results: Five to eight days after the first dose of LDERC, median adrenocorticotropic hormone (ACTH) concentration was lower (p = 0.001), changing from 153 pg/mL (IQR: 78, 331) to 57 pg/mL (IQR: 30, 102). With HDERC, median ACTH concentration was also 153 pg/mL (IQR: 96, 185) before and then 56 pg/mL (IQR: 29, 86) after 5–8 days of treatment (p = 0.047). Over 12 months of treatment, ACTH concentration ranged from 14 to >1,250 pg/mL (median: 51 pg/mL) in horses treated with LDERC and 20 to 472 pg/mL (median: 50 pg/mL) in horses treated with HDERC. Measurements remained above the seasonal reference range in 39.3 and 52.3% of horses treated with LDERC and HDERC, respectively. Clinical improvement was reported by owners in 78.3 and 100% of horses treated with LDERC and HDERC, respectively. Partial, self-limiting inappetence was reported in 30.0% of LDERC and 60% HDERC cases. Seven horses exhibited lethargy (5 LDERC, 2 HDERC). Insulin concentrations measured 30 days post-ERC treatment were no different from baseline.

Discussion: Clinical and endocrinological responses were consistent with results of previous reports of oral pergolide treatment. Weekly injection of ERC may be an effective alternative to pergolide; the 0.005 mg/kg dose appeared to be as effective, with less risk of inappetence, than the 0.01 mg/kg dose that has been reported previously.

Laminitis associated with hyperinsulinaemia is a significant cause of morbidity and mortality in horses with equine metabolic syndrome. The diagnosis and management of hyperinsulinaemia are therefore critical to prevent the development of laminitis. This review article aims to help primary care clinicians manage patients with hyperinsulinaemia by providing an overview of diagnostics, management strategies and new therapies that are available.