Metabolic modeling using statistical and spreadsheet software: Application to the glucose minimal model.

Kinetic non-linear metabolic models are used extensively in medical research and increasingly for clinical diagnostic purposes. An example of such a model is the Glucose Minimal Model by Bergman and colleagues [1]. This model is similar to pharmacokinetic/pharmacodynamic models in that like pharmacokinetic/pharmacodynamic models, it is based on a small number of fairly simple ordinary differential equations and it aims to determine how the changing concentration of one blood constituent influences the concentration of another constituent. Although such models may appear prima facie, to be relatively simple, they have gained a reputation of being difficult to fit to data, especially in a consistent and repeatable fashion. Consequently, researchers and clinicians have generally relied on dedicated software packages to do this type of modeling. This article describes the use of statistical and spreadsheet software for fitting the Glucose Minimal Model to data from an insulin modified intravenous glucose tolerance test (IM-IVGTT). A novel aspect of the modeling is that the differential equations that are normally used to describe insulin action and the disposition of plasma glucose are first solved and expressed in their explicit forms so as to facilitate the estimation of Glucose Minimal Model parameters using the nonlinear (nl) optimization procedure within statistical and spreadsheet software. The most important clinical parameter obtained from the Glucose Minimal Model is insulin sensitivity (SI). Using IM-IVGTT data from 42 horses in one experiment and 48 horses in a second experiment, we demonstrate that estimates of SI derived from the Glucose Minimal Model fitted to data using STATA and Excel, are highly concordant with SI estimates obtained using the industry standard software, MinMod Millennium. This work demonstrates that there is potential for statistical and spreadsheet software to be applied to a wide range of kinetic non-linear modeling problems.

A multifaceted analytical approach for detecting effects on semen quality when using small sample sizes.

Driven by technical, logistical and economic limitations, detection of treatment effects on semen quality typically include the design and collection of small sample datasets. A consequence of these small sample studies is that they suffer low statistical power. Historically, researchers faced with small sample size studies have relied upon non-parametric analysis; however, this approach is still unlikely to tease out a true statistical significance based upon limited sample size. Here we propose a novel methodology that can be applied in small samples study situations that combines repeated measures ANOVA and Mixed-Effects linear regression models with Bayesian Linear regression modeling when evaluating for treatment effects on quantitative semen quality parameters. Using this methodology, we show that investigating the data with this multifaceted analytical technique results in improved reproducibility and sensitivity of the findings while minimizing the likelihood of Type 1 errors when combining the inference statistics from multiple models/methodologies using Bayes Factor analysis.

Effects of dry period energy intake on insulin resistance, metabolic adaptation, and production responses in transition dairy cows on grass silage-based diets.

High energy intake in the dry period has reportedly had adverse effects on mobilization of body reserves, dry matter intake, and productivity of dairy cows. We investigated whether grass silage (GS) fed ad libitum (high energy intake, HEI; 141% of daily metabolizable energy requirements) in an 8-wk dry period affects metabolic adaptation-specifically, peripheral insulin resistance-compared with a total mixed ration consisting of GS, wheat straw, and rapeseed meal (55/40/5%; controlled energy intake, CEI; 108% of metabolizable energy/d) fed ad libitum. Multiparous Ayrshire dairy cows (n = 16) were used in a randomized complete block design until 8 wk after parturition. Commercial concentrates were fed 1 and 2 kg/d during the last 10 to 6 and 5 to 0 d before the expected calving date, respectively. Postpartum, a similar lactation diet with ad libitum access to GS and increasing concentrate allowance (maximum of 16 kg/d) was offered to all. The HEI group gained more body weight and had higher plasma insulin, glucose, and ß-hydroxybutyrate concentrations than the CEI group prepartum. Postpartal plasma glucose tended to be higher and milk yield was greater from wk 5 onward for HEI compared with CEI cows. An intravenous glucose tolerance test (IVGTT) was performed at -13 ± 5 d and 9 ± 1 d relative to calving. The HEI cows had greater insulin response to glucose load and smaller area under the response curve for glucose than CEI cows in prepartal IVGTT. Thus, compensatory insulin secretion adapted to changes in insulin sensitivity of the peripheral tissues, preserving glucose tolerance of HEI cows. Higher insulin levels were needed in HEI cows than in CEI cows to elicit a similar decrement of nonesterified fatty acid concentration in prepartal IVGTT, suggesting reduced inhibition of lipolysis by insulin in HEI cows before parturition. In conclusion, high energy intake of moderately digestible GS with low concentrate feeding in the close-up dry period did not have adverse effects on metabolic adaptation, insulin sensitivity, and body mobilization after parturition. Instead, this feeding regimen was more beneficial to early-lactation performance than GS-based total mixed ration diluted with wheat straw.

Pharmacokinetics, disposition, and plasma concentrations of dimethyl sulfoxide (DMSO) in the horse following topical, oral, and intravenous administration.

Compartmental models were used to investigate the pharmacokinetics of intravenous (i.v.), oral (p.o.), and topical (TOP) administration of dimethyl sulfoxide (DMSO). The plasma concentration-time curve following a 15-min i.v. infusion of DMSO was described by a two-compartment model. Median and range of alpha (t1/2α ) and beta (t1/2ß ) half-lives were 0.029 (0.026-0.093) and 14.1 (6.6-16.4) hr, respectively. Plasma concentration-time curves of DMSO following p.o. and TOP administration were best described by one-compartment absorption and elimination models. Following the p.o. administration, median absorption (t1/2ab ) and elimination (t1/2e ) half-lives were 0.15 (0.01-0.77) and 15.5 (8.5-25.2) hr, respectively. The plasma concentrations of DMSO were 47.4-129.9 µg/ml, occurring between 15 min and 4 hr. The fractional absorption (F) during a 24-hr period was 47.4 (22.7-98.1)%. Following TOP administrations, the median t1/2ab and t1/2e were 1.2 (0.49-2.3) and 4.5 (2.1-11.0) hr, respectively. Plasma concentrations were 1.2-8.2 µg/ml occurring at 2-4 hr. Fractional absorption following TOP administration was 0.48 (0.315-4.4)% of the dose administered. Clearance (Cl) of DMSO following the i.v. administration was 3.2 (2.2-6.7) ml hr-1  kg-1 . The corrected clearances (ClF ) for p.o. and TOP administrations were 2.9 (1.1-5.5) and 4.5 (0.52-18.2) ml hr-1  kg-1 .

Overground endoscopy in 311 Thoroughbred racehorses: findings and correlation to resting laryngeal function.

OBJECTIVE: To review a large number of equine overground endoscopy (OGE) examinations to determine the incidence of dynamic upper airway obstructions (DUAO); correlations were explored with laryngeal endoscopy findings at rest and abnormal exercising respiratory noise. METHODS: Retrospective analysis of horses presenting for OGE because of perceived poor performance and/or history of abnormal exercising respiratory noise between 2010 and 2014. Signalment, history and examination findings during resting laryngeal endoscopy and OGE were reviewed. RESULTS: Of the total examinations, 311 were reviewed. One or more DUAO were found in 249/311 horses. From 210 males (colts and geldings), 121 had arytenoid cartilage collapse (ACC) and 111 had vocal fold collapse (VFC). From 101 females, 25 had intermittent dorsal displacement of the soft palate (DDSP). Resting laryngeal function grade 4 was found in 121/311 of the study population and 92/210 of males. An association was found between horses with lower resting arytenoid abduction ability to dynamic ACC and higher resting arytenoid abduction ability with DDSP. Abnormal exercising respiratory noise was positively associated with the presence of DUAO. CONCLUSIONS: Multiple DUAO in association with abnormal exercising respiratory noise was a common finding in horses examined for poor performance. This study highlights the importance of OGE in accurately diagnosing the nature of DUAO associated with poor performance.

Effects of dietary energy allowance and decline in dry matter intake during the dry period on responses to glucose and insulin in transition dairy cows.

We assessed whether high energy intake during the early dry period [144% of metabolizable energy (ME) requirements/d] followed by a gradual restriction of energy intake in the close-up dry period (119% of ME/d; HEI) impaired whole-body insulin sensitivity compared with a controlled energy intake (100% of ME/d; CEI) throughout the 6-wk dry period. Multiparous Ayrshire dairy cows (n = 16) were blocked by body weight, body condition score, and expected date of parturition and were used in a randomized complete block design until 10 d after parturition. Cows were fed either HEI or CEI diets based on grass silage during the first 3 wk of the dry period and grass silage supplemented with a commercial concentrate (30% of ME intake) during the final 3 wk of gestation. After calving, all cows were fed grass silage ad libitum and an increasing amount of commercial concentrate (maximum 9 kg at d 10 postpartum). Intravenous glucose tolerance tests (IVGTT) and intravenous insulin challenges were performed -10 ± 5 d (n = 15) and +10 ± 1 d (n = 14) relative to parturition. Following glucose injection, we did not find any treatment effects on glucose and insulin responses. The prepartal nonesterified fatty acid (NEFA) response of the HEI group was blunted, basal NEFA and the decrement of NEFA were smaller, and the area under the response curve (AUC) of NEFA was less negative in HEI cows than in CEI cows. The NEFA response reversed after parturition; the NEFA AUC of the HEI group was more negative than that of the CEI group. We did not find similar responses after insulin injection. Across the treatments, NEFA AUC correlated strongly with the basal NEFA concentration during the IVGTT pre- and postpartum. Calculated and model-based indices characterizing the overall glucose tolerance and ß-cell function and the insulin sensitivity were higher after parturition than during the dry period. Consistent with the lower basal insulin, the acute insulin release after the glucose infusion was smaller in postpartal IVGTT than in prepartal IVGTT. The results suggest that whole-body insulin sensitivity of the cows increased after parturition. However, the role of peripheral insulin sensitivity in the regulation of glucose partitioning seems to be minor relative to the major change in insulin secretion and clearance during the periparturient period.

Determining insulin sensitivity from glucose tolerance tests in sheep.

A mathematical model of the dynamics of insulin and glucose during a frequently sampled intravenous glucose tolerance test (IVGTT) in sheep was developed that characterizes the large second-phase insulin secretion response in sheep during IVGTT. The model was fit to measurements of the glucose and insulin dynamics during standard IVGTT ( = 42) and modified IVGTT ( = 40), where insulin was injected 60 min after the initiation of the IVGTT. The correlation between log insulin sensitivity determined by hyperglycemic clamps (HGC) and standard IVGTT was = 0.43 ( = 0.005). The correlation between log insulin sensitivity determined by HGC and modified IVGTT was = 0.51 ( = 0.002). The model, therefore, provides a method to determine insulin sensitivity through a cheaper and more easily performed IVGTT. We validated our estimation procedure using 2 independent experiments on the effect of 1) pregnancy and 2) being born preterm and exposed to dextrose or dextrose with insulin on HGC-derived insulin sensitivity. The IVGTT-derived insulin sensitivity was significantly greater in pregnant ewes than in prepregnant ewes (difference of 0.39 ± 0.12 log n ng mL; < 0.05), and this was consistent with the significantly greater hyperinsulinemic euglycemic clamp-derived insulin sensitivity in pregnant ewes than in prepregnant ewes (difference of 4.03 ± 0.66 µmol mL kg min ng; < 0.001). There was no significant effect of being born preterm on IVGTT/HGC-derived insulin sensitivity. Basal insulin, insulin sensitivity, insulin production, and insulin clearance were lower in prepregnant ewes ( < 0.05). That is, prepregnant ewes have a lower insulin equilibrium status and less responsive insulin turnover. There was also a significant effect of insulin therapy on the rate of insulin clearance in preterm lambs ( < 0.05). This effect was independently significant of its covariance with all other model parameters. Therefore, it can be interpreted as a direct effect on the rate of insulin clearance by the insulin treatment. All other parameter responses to the insulin treatment effect can be regarded as being due to the covariance between these parameters. These analyses demonstrate that treatment effects on insulin sensitivity can be detected using IVGTT experiments.

Streptococcus equi Detection Polymerase Chain Reaction Assay for Equine Nasopharyngeal and Guttural Pouch Wash Samples.

BACKGROUND: Bacterial culture and polymerase chain reaction (PCR) assays for the detection of Streptococcus equi in nasopharyngeal washes (NPW) and guttural pouch lavage (GPL) samples have low sensitivity. In human diagnostics, processing of samples with flocked swabs has improved recovery rates of bacterial agents because of improved surface area and elution factors. HYPOTHESIS: For S. equi subsp. equi (S. equi) detection in NPW and GPL samples we hypothesized that: direct-PCR would be more reliable than flocked swab culture (FS culture); flocked swab PCR (FS-PCR) would be equivalent to direct-PCR; and FS culture would be more reliable than traditional culture. SAMPLES: A total of 193 samples (134 NPW and 59 GPL) from 113 horses with either suspected S. equi infection, convalescing from a known S. equi infection, or asymptomatic horses screened for S. equi. METHODS: Prospective study. Samples were submitted for S. equi direct-PCR. Using logistic regression, direct-PCR (gold standard) was compared to FS culture, traditional culture, and FS-PCR also performed. RESULTS: Direct-PCR was statistically more sensitive than FS-PCR, FS culture, and traditional culture (P < .001). All methods had sensitivities <70% relative to the direct-PCR. FS culture had a similar sensitivity relative to traditional culture. The odds of GPL samples being positive on direct-PCR (P = .030) and FS-PCR were greater than those for NPW samples (P = .021). CONCLUSIONS AND CLINICAL IMPORTANCE: Use of flocked swabs during laboratory preprocessing did not improve detection of S. equi via either PCR or bacterial culture from samples. Direct-PCR is the preferred method of detection of S. equi.

Survival Time of Cross-Match Incompatible Red Blood Cells in Adult Horses.

BACKGROUND: There is a markedly reduced half-life of transfused RBCs when donor and recipient cats or humans are cross-match incompatible. Only 10-20% of horses have naturally occurring alloantibodies. Therefore, cross-match testing before blood transfusion is not always performed. HYPOTHESIS: Cross-match incompatibility predicts shortened RBC survival time as compared to that of compatible or autologous blood. ANIMALS: Twenty healthy adult horses. METHODS: Prospective trial. Blood type, anti-RBC antibody screen (before and 1 month after transfusion) and major and minor cross-match determined 10 donor-recipient pairs. Two pairs were cross-match compatible, the remainder incompatible. Donor blood (4 L) was collected into citrate phosphate dextrose adenine-1, labeled with NHS-biotin, and transfused into recipients. Samples were collected at 1 hour and 1, 2, 3, 5, 7, 14, 21, 28, and 35 days after transfusion, and biotinylated RBCs were detected by flow cytometry. Horses were monitored for transfusion reaction during transfusion and daily for 5 days. RESULTS: Cross-match incompatibility was significantly associated with decreased RBC survival time (P < .001). The half-life of transfused incompatible (cross-match >1+) allogenic equine RBCs was 4.7 (95% CI, 3.2-6.2) days versus 33.5 (24-43) days for compatible pairings. Cross-match incompatibility was associated with acute febrile transfusion reaction (P = .0083). At day 30, only 1 horse had developed novel anti-RBC antibodies. CONCLUSIONS AND CLINICAL IMPORTANCE: Cross-match incompatibility was predictive of febrile transfusion reaction and shortened transfused RBC survival, but did not result in production of anti-RBC antibodies at 30 days. Cross-match testing before transfusion is recommended.

Plasma Peak and Trough Gentamicin Concentrations in Hospitalized Horses Receiving Intravenously Administered Gentamicin.

BACKGROUND: Gentamicin is an aminoglycoside antimicrobial commonly used in horses at 6.6 mg/kg IV once daily. Therapeutic drug monitoring (TDM) can confirm desired peak concentration is reached for common bacterial isolates, and detect toxicosis associated with high trough values. OBJECTIVES: Determine the relationship between gentamicin dose and plasma concentration in hospitalized horses, and identify a starting dose range to achieve peaks > 32 µg/mL. ANIMALS: Sixty-five horses (2002-2010) receiving once-daily gentamicin with TDM performed (N = 99 sets). METHODS: Retrospective study. Data from hospitalized horses including weight, dose, plasma peak, and trough gentamicin concentration, creatinine concentrations and presence of focal or systemic disease were collected from medical records. Peak concentrations measured 25-35 minutes after administration were included (N = 77). Data were divided into low (<7.7 mg/kg), medium (7.7-9.7 mg/kg) and high (>9.7 mg/kg) dose groups, and were grouped by the horse having focal or systemic disease. RESULTS: Peak concentrations resulting from doses ≥7.7 mg/kg were 5.74 µg/mL (SE 2.1 µg/mL) greater than peaks from doses <7.7 mg/kg (P = .007). Peak concentrations was 3.6 times more likely to be >32 µg/mL if dose was ≥7.7 mg/kg (P = .04). There were no significant effects of dose on trough or creatinine concentration. At a given dose, horses with focal disease had higher peaks than those with systemic disease (P = .039). CONCLUSIONS AND CLINICAL IMPORTANCE: These data suggest gentamicin dosage should be individually determined in horses using TDM, but support an initial once-daily dose of 7.7-9.7 mg/kg IV to achieve peaks >32 µg/mL and trough concentrations <2 µg/mL. Further studies evaluating the safety of doses >6.6 mg/kg are required.

The Use of Recombinant Tissue Plasminogen Activator (rTPA) in The Treatment of Fibrinous Pleuropneumonia in Horses: 25 Cases (2007-2012).

BACKGROUND: Information about treatment protocols, adverse effects and outcomes with intrapleural recombinant tissue plasminogen activator (rTPA) use in horses with fibrinous pleuropneumonia is limited. HYPOTHESIS/OBJECTIVES: Describe factors that contribute to clinical response and survival of horses treated with rTPA intrapleurally. ANIMALS: Horses with bacterial pneumonia and fibrinous pleural effusion diagnosed by ultrasonography, that were treated with rTPA intrapleurally. METHODS: Retrospective multicenter case series from 2007-2012. Signalment, history, clinical and laboratory evaluation, treatment, and outcome obtained from medical records. Regression analysis used to identify associations between treatments and outcomes. RESULTS: Thirty three hemithoraces were treated in 25 horses, with 55 separate treatments. Recombinant tissue plasminogen activator (375-20,000 µg/hemithorax) was administered 1-4 times. Sonographically visible reduction in fibrin mat thickness, loculations, fluid depth, or some combination of these was seen in 32/49 (65%) treatments. Response to at least 1 treatment was seen in 17/20 (85%) horses with sonographic follow-up evaluation after every treatment. Earlier onset of rTPA treatment associated with increased survival odds. No association was found between cumulative rTPA dose or number of rTPA doses and survival, development of complications, duration of hospitalization or total charges. Clinical evidence of hypocoagulability or bleeding was not observed. Eighteen horses (72%) survived to discharge. CONCLUSIONS AND CLINICAL IMPORTANCE: Treatment with rTPA appeared safe and resulted in variable changes in fibrin quantity and organization within the pleural space. Recombinant tissue plasminogen activator could be a useful adjunct to standard treatment of fibrinous pleuropneumonia, but optimal case selection and dosing regimen remain to be elucidated.

The Association of Fibrinous Pleural Effusion with Survival and Complications in Horses with Pleuropneumonia (2002-2012): 74 Cases.

BACKGROUND: Fibrinous parapneumonic pleural effusions are associated with decreased efficacy of pleural fluid drainage and increased risk of medical treatment failure in people, but similar associations have not been established in horses. HYPOTHESIS/OBJECTIVES: We hypothesized that fibrin deposition in the pleural cavity of horses with parapneumonic effusions increases the risk of poor outcome. ANIMALS: Seventy four horses with bacterial pleuropneumonia diagnosed by culture and cytology of tracheal aspirates, pleural fluid, or both, and pleural effusion diagnosed by ultrasonographic examination. METHODS: Retrospective study of cases was from 2002 to 2012. Information obtained from the medical records included signalment, history, sonographic findings, treatments, and outcome. The primary outcome investigated was survival and secondary outcomes were development of complications and surgical intervention. Fisher's exact test and logistic regression were applied for categorical variables. A t-test was used to find differences in continuous variables between groups. RESULTS: Seventy four horses met study criteria and 50 (68%) survived. Fibrinous pleural effusion was associated with higher respiratory rate and pleural fluid height at admission, necrotizing pneumonia, increased number of indwelling thoracic drains required for treatment, and decreased survival. CONCLUSIONS AND CLINICAL IMPORTANCE: Fibrin accumulation in parapneumonic effusions is associated with increased mortality. Direct fibrinolytic treatment might be indicated in affected horses.

Relationship of adiponectin and its multimers to metabolic indices in cats during weight change.

Adiponectin is an important anti-inflammatory hormone secreted from adipose tissue. The high-molecular-weight form of adiponectin (HMW) closely correlates with insulin sensitivity in human beings. This study uses a novel method of size-exclusion gel chromatography combined with enzyme-linked immunosorbent assay to measure HMW feline adiponectin and determine its relationship to leptin, cholesterol, and insulin sensitivity as cats gain and lose weight. In addition, total adiponectin and its messenger RNA expression in subcutaneous adipose tissue were measured. No correlations were found between total serum adiponectin and subcutaneous adipose messenger RNA expression, fat mass, or measures of insulin sensitivity. This study demonstrates that cats have high percentages of HMW adiponectin. Although weak correlations between HMW adiponectin and fat mass were detected, additional cats are needed to determine if the correlations are significant.

Experimental hyperlipidemia induces insulin resistance in sheep.

This study aimed to evaluate the effects of intravenous infusion of a soybean-based lipid emulsion on some blood energy-related metabolites and insulin sensitivity indexes in sheep. Four clinically healthy ewes were assigned into a 2-treatment, 2-period cross-over design. Either normal saline (NS) or lipid emulsion (LE) was intravenously introduced at a rate of 0.025 mL·kg(-1) min(-1) for 6 h. The concentrations of blood nonesterified fatty acid (NEFA), beta-hydroxybutyrate, triglyceride, cholesterol, urea, creatinine, cortisol, glucose, and insulin were measured at different time points. After 6 h, intravenous glucose tolerance test was performed. Lipid infusion elicited an increase (P < 0.05) in the NEFA, beta-hydroxybutyrate, and triglyceride concentrations compared with the baseline value and NS infusion. Infusion of NS did not influence blood glucose concentration; however, LE infusion increased plasma glucose concentration (P < 0.05). At time point 12 h, serum insulin concentrations were increased (P < 0.05) in NS treatment; however, such an increase was not observed in the LE treatment. Insulin sensitivity index for the LE infusion was lower (P < 0.05) than that for the NS treatment. The glucose effectiveness was not (P > 0.05) different among treatments. In the LE treatment, acute-phase insulin responses increased (P < 0.05) and disposition index decreased (P < 0.001) compared with NS treatment. The results showed that experimentally induced NEFA in blood could cause insulin resistance in sheep. The current model could be used to evaluate the pathogenesis of conditions associated with increased lipid mobilization and insulin resistance.

Preliminary investigation of the area under the L-lactate concentration-time curve (LACArea) in critically ill equine neonates.

BACKGROUND: A variety of measures of L-lactate concentration ([LAC]) in the blood of critically ill neonatal foals have shown utility as prognostic indicators. These measures, evaluating either the severity of hyperlactatemia or the duration of exposure to hyperlactatemia, perform fairly well and have correctly classified 75-80% of foals examined in several studies. The area under the L-lactate concentration versus time curve (LACArea) encompasses both severity and duration of hyperlactatemia and should improve correct classification of patient survival. HYPOTHESIS/OBJECTIVES: LACArea is larger in nonsurviving critically ill neonatal foals. ANIMALS: Forty-nine foals admitted for critical illness to 1 of 4 referral hospitals. METHODS: Whole blood was obtained at admission and 6, 12, 18, and 24 hours after admission for measurement of L-lactate using a handheld lactate meter. LACArea was calculated for: admission-6, 6-12, 12-18, 18-24 hours, and admission-24 hours using the trapezoidal method and summing the 6-hours interval areas to determine total 24 hours area. Differences between survivors and nonsurvivors were determined using robust regression and Kruskal-Wallis testing, P < .05. RESULTS: LACArea was significantly larger in nonsurviving foals (n = 9) than in surviving foals (n = 40) at all time periods examined. CONCLUSIONS AND CLINICAL IMPORTANCE: Differences in LACArea between surviving and nonsurviving critically ill neonatal foals are large and support further investigation of this method as an improved biomarker for survival in critically ill neonatal foals is indicated.

Analysis of stress fractures associated with lameness in Thoroughbred flat racehorses training on different track surfaces undergoing nuclear scintigraphic examination.

REASONS FOR PERFORMING STUDY: There is limited information regarding the impact of training track surface on the occurrence of stress fractures. OBJECTIVES: To evaluate the impact of training track surface on the proportion of long bone and pelvic stress fractures associated with lameness in Thoroughbred horses in flat race training undergoing nuclear scintigraphic examination. STUDY DESIGN: Retrospective study. METHODS: Scintigraphic examinations of Thoroughbred flat racehorses were evaluated from 2 hospitals (hospital A [Toronto Equine Hospital], 2003-2009, and hospital B [George D. Widener Hospital for Large Animals, School of Veterinary Medicine, University of Pennsylvania], 1994-2006). Horses admitted to hospital A trained at a single track, at which the main training surface changed from dirt to synthetic on 27 August 2006. Two distinct populations existed at hospital B: horses that trained on dirt (numerous trainers) and those that trained on turf (single trainer). All scintigraphic images were evaluated by a blinded reviewer. Fisher's exact test and logistic regression were used when appropriate, and significance was set at P<0.05. RESULTS: When reviewing 528 scintigraphic examinations from hospital A (257 dirt and 271 synthetic, numerous trainers), there was a greater proportion of stress fractures detected in scintigraphic examinations from horses training on a synthetic surface (31.7%) in comparison to scintigraphic examinations from horses training on a dirt surface (23.0%) at an earlier point in time (P = 0.03). There was a greater proportion of hindlimb/pelvic and tibial stress fractures diagnosed in horses from the synthetic surface-trained group than from the dirt-trained group at hospital A (P<0.04 and P = 0.03, respectively). CONCLUSIONS: This study provides evidence that training surface may affect the proportion of stress fractures diagnosed, but other factors, such as training philosophy, appear to be important. Future prospective investigations to fully elucidate the relationship between training track surface and the proportion of stress fractures and other nonfatal musculoskeletal injuries are warranted.

Occurrence of cardiac arrhythmias in Standardbred racehorses.

REASONS FOR PERFORMING STUDY: Cardiac arrhythmias are a recognised but poorly characterised problem in the Standardbred racehorse. Frequency data could aid the development of cardiac arrhythmia screening programmes. OBJECTIVES: To characterise the occurrence of cardiac arrhythmias in Standardbreds prior to racing and in the late post race period using a handheld, noncontinuous recording device. STUDY DESIGN: Prospective, observational study, convenience sampling. METHODS: Noncontinuous electrocardiographic recordings were obtained over a 12 week period from Standardbred horses competing at a single racetrack. Electrocardiograms were obtained before racing and between 6 and 29 min after the race using a handheld recording device. Prevalence of arrhythmias was calculated for all horses and overall frequency of arrhythmias was calculated for race starts and poor performers. Univariate logistic regression analysis was used to identify risk factors for cardiac arrhythmias. RESULTS: A total of 8657 electrocardiogram recordings were obtained from 1816 horses. Six horses had atrial fibrillation after racing (prevalence = 0.11%, frequency = 0.14%), one horse had supraventricular tachycardia before racing (prevalence = 0.06%, frequency = 0.02%), and 2 horses had ventricular tachyarrhythmias after racing (prevalence = 0.06%, frequency = 0.05%). The frequency of atrial fibrillation among race starts with poor performance was 1.3-2.0%. Increasing age was a significant risk factor for the presence of atrial premature contractions before racing and atrial fibrillation and ventricular ectopy after racing. CONCLUSIONS: Both physiological and pathological cardiac arrhythmias can be detected in apparently healthy Standardbred horses in the prerace and late post race period using noncontinuous recording methods. Future studies should examine cumulative training or racing hours as a risk factor for cardiac arrhythmia. The prevalence and frequency information may be useful for track veterinarians and regulatory personnel following trends in cardiac arrhythmias.

Sequential L-lactate concentration in hospitalised equine neonates: A prospective multicentre study.

REASONS FOR PERFORMING STUDY: Evaluation of serial blood lactate concentrations [LAC] are of prognostic value for morbidity and mortality in critically ill human patients and neonatal foals, but have not been prospectively evaluated in a large multicentre study of critically ill neonatal foals. OBJECTIVES: To prospectively evaluate the prognostic value of sequential [LAC] analysis in critically ill neonatal foals with risk of mortality. STUDY DESIGN: Prospective, observational study. METHODS: Thirteen university and private equine referral hospitals enrolled 643 foals over the 2008 foaling season and [LAC] was measured at admission ([LAC]ADMIT ) and 24 ([LAC]24 ), 48 ([LAC]48 ), 72 ([LAC]72 ), 96 ([LAC]96 ) and 120 h ([LAC]120 ) after admission. [LAC] changes over time ([LAC]Δ) were calculated between sampling points. RESULTS: Nonsurvivors had significantly greater [LAC]ADMIT , [LAC]24 and [LAC]48 compared with surviving foals (P<0.001). In nonsurviving foals [LAC]Δ did not decrease over time while survivors showed significant positive [LAC]Δ between [LAC]ADM -24 and all other time periods (P<0.001). Logistic regression analysis showed that the odds of survival decreased for each 1 mmol/l [LAC] increase at all time points for all critically ill foals, independent of major final diagnoses as potential confounders. Septic foals had significantly greater [LAC] at all time points compared with nonseptic foals (P<0.001) and [LAC]Δ in septic foals was significantly more positive (suggesting better clearance of lactate from the blood) only at [LAC]ADM -24 and [LAC]72-96 (P<0.01), while in nonseptic foals [LAC]Δ was significantly positive between [LAC]ADM -24 compared with all other time periods (P<0.001). CONCLUSIONS: Blood lactate concentration is a strong, independent biomarker used to predict mortality in critically ill foals. Lactate metabolism is impaired in nonsurviving and septic foals and [LAC]Δ can be utilised to identify patients at high risk for mortality.

Diurnal patterns of grazing behavior and humoral factors in supplemented dairy cows.

Offering feed supplements to grazing dairy cows results in a reduction in grazing time. However, the effect differs depending on the time of day that feeds are offered. To understand the physiological basis for this, associations among circulating factors known to be associated with intake regulation in monogastric species and grazing behavior in the dairy cow were investigated. Seventeen multiparous cows at 28 ± 5 d in milk grazed together and consumed 4.4 kg of dry matter/d of a pelleted concentrate feed supplement, equally split, at the a.m. and p.m. milking. Grazing behavior was recorded over 4 consecutive days in all 17 cows. Blood was sampled from 10 of the 17 cows every 4h over a 48-h period following the grazing behavior measurements; sampling times were staggered by 2h to provide a diurnal profile of humoral factors. Grazing profiles illustrated major grazing bouts after the a.m. and p.m. milking; however, the p.m. grazing bout was characterized as the most intensive and time spent grazing was unaffected by supplementation. Associations among proportion of cows grazing and circulating hormones and metabolites differed throughout the day. During the a.m., relationships were consistent with those reported in monogastric species, with ghrelin and nonesterified fatty acids decreasing and insulin increasing with feeding. In comparison, during the major grazing bout predusk, ghrelin concentrations increased until sunset, despite the large proportion of cows grazing, before declining; this is consistent with ghrelin stimulating the predusk grazing bout. Results indicate that humoral factors known to affect hunger and satiety in monogastric animals may also have a potential role in the physiological regulation of diurnal and feeding behavior in ruminants.

Comparison of force plate gait analysis and owner assessment of pain using the Canine Brief Pain Inventory in dogs with osteoarthritis.

BACKGROUND: Lameness assessment using force plate gait analysis (FPGA) and owner assessment of chronic pain using the Canine Brief Pain Inventory (CBPI) are valid and reliable methods of evaluating canine osteoarthritis. There are no studies comparing these 2 outcome measures. OBJECTIVE: Evaluate the relationship between CBPI pain severity (PS) and interference (PI) scores with the vertical forces of FPGA as efficacy measures in canine osteoarthritis. ANIMALS: Sixty-eight client-owned dogs with osteoarthritis (50 hind limb and 18 forelimb). METHODS: Double-blind, randomized. Owners completed the CBPI, and dogs underwent FPGA on days 0 and 14. Dogs received carprofen or placebo on days 1 through 14. The change in PS and PI scores from day 0 to 14 were compared to the change in peak vertical force (PVF) and vertical impulse (VI). RESULTS: PS and PI scores significantly decreased in carprofen- compared with placebo-treated dogs (P = .002 and P = .03, respectively). PVF and VI significantly increased in carprofen- compared with placebo-treated dogs (P = .006 and P = .02, respectively). There was no correlation or concordance between the PS or PI score changes and change in PVF or VI. CONCLUSIONS AND CLINICAL IMPORTANCE: In these dogs with hind limb or forelimb osteoarthritis, owner assessment of chronic pain using the CBPI and assessment of lameness using FPGA detected significant improvement in dogs treated with carprofen. The lack of correlation or concordance between the change in owner scores and vertical forces suggests that owners were focused on behaviors other than lameness when making efficacy evaluations in their dogs.