OBJECTIVE To compare the degree of white line separation created by increasing physiologic loads between bovine claws with and without toe-tip necrosis (TTN). SAMPLE Cadaveric bovine hind limbs with (n = 10) and without (10) TTN. PROCEDURES Hind limbs in which 1 or both claws had evidence of apical white line separation were considered to have TTN. Hind limbs in which neither claw had evidence of white line separation were considered controls. Each hind limb was mounted in a materials testing system with the bottom surface of the hoof angled at approximately 5° to the horizontal plane such that the apex of the claws made initial contact with the clear testing surface to simulate physiologic loading conditions. A digital camera mounted underneath the testing surface was used to obtain images of the bottom of the hoof during the application of each of 3 increasing static loads (1, 2, and 3 kN). The images were analyzed with commercial image-processing software to quantify white line separation area. RESULTS White line separation area was significantly greater for claws with TTN than for control claws and increased as the applied load increased. White line separation was almost nonexistent in control claws and was not affected by increasing load. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that mechanical loading exacerbated TTN, but compressive loading alone, even at excessive levels, did not initiate the condition. Interventions (eg, hoof blocks) that decrease loading of affected claws may be beneficial for the treatment of TTN at its earliest stages.

OBJECTIVE To biomechanically assess outcomes for 2 percutaneous gastropexy techniques and determine the amount of time necessary to perform the techniques for securing percutaneous endoscopic gastrostomy (PEG) tubes in canine cadavers. ANIMALS 18 canine cadavers. PROCEDURES 6 cadavers were assigned to each of 3 groups (PEG tube only, PEG tube with T-fastener gastropexy, and PEG tube with U-stitch gastropexy). Time to completion of placement of a PEG tube and gastropexy was recorded. After tubes were placed, the stomach and body wall on the left side of the abdomen were removed and biomechanically tested. Maximum tension at the time of initial failure of the PEG tube or gastropexy was recorded. RESULTS Significantly more force was required to induce failure for the T-fastener and U-stitch techniques than for the PEG tube only technique. In addition, both the T-fastener and U-stitch techniques required significantly more time for placement than did the PEG tube only technique. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that performing a T-fastener or U-stitch gastropexy may decrease the risk of early dislodgement of a PEG tube in dogs. However, studies conducted with these techniques in live dogs would be necessary to confirm this hypothesis. The additional amount of time needed to perform the T-fastener or U-stitch gastropexy would likely be clinically unimportant.

OBJECTIVE To assess effects of catheter diameter and injection rate of flush solution (saline [0.9% NaCl] solution) on renal contrast-enhanced ultrasonography (CEUS) with perfluorobutane in dogs. ANIMALS 5 healthy Beagles. PROCEDURES CEUS of the kidneys was performed by IV injection of contrast medium (0.0125 mL/kg) followed by injection of 5 mL of saline solution at rates of 1, 3, and 5 mL/s through a 20-gauge or 24-gauge catheter; thus, CEUS was repeated 3 times for each catheter diameter. Time-intensity curves were created for regions of interest drawn in the renal cortex and medulla. Repeatability was determined by calculating the coefficient of variation (CV). Statistical analysis was used to assess whether perfusion variables or CV of the perfusion variables was associated with catheter diameter or injection rate. RESULTS Perfusion variables did not differ significantly between catheter diameters. Time to peak enhancement (TTP) in the renal cortex was affected by injection rate, and there were significantly lower values for TTP at higher injection rates. The CEUS variables with the lowest CVs among injection rates were TTP for the renal cortex; the CV for TTP of the renal cortex was the lowest at an injection rate of 5 mL/s. CONCLUSIONS AND CLINICAL RELEVANCE Use of a 24-gauge catheter did not alter CEUS with perfluorobutane; therefore, such catheters could be used for CEUS of the kidneys of small dogs. Moreover, a rate of 5 mL/s is recommended for injection of flush solution to obtain greater accuracy for renal CEUS in Beagles.

OBJECTIVE To evaluate coagulation factors in units of leukoreduced (LR) and nonleukoreduced (non-LR) canine fresh-frozen plasma (cFFP). ANIMALS 8 healthy research dogs. PROCEDURES In a crossover study, dogs were randomly assigned to 1 of 2 groups from which blood was collected and either did or did not undergo leukoreduction. After a recovery period of ≥ 28 days, the dogs were switched between protocols. After each collection, blood samples were centrifuged, and cFFP was stored frozen for later comparative analysis of coagulation factors, antithrombin, and protein C activities (reported as comparative percentages of the corresponding activities determined in a canine pooled plasma standard); prothrombin and activated partial thromboplastin times; and fibrinogen concentration. RESULTS There were no significant differences detected between results for LR cFFP, compared with those for non-LR cFFP. CONCLUSIONS AND CLINICAL RELEVANCE Although there was variation among residual activities of coagulation factors in LR and non-LR cFFP, the variations and differences were considered unlikely to impact the efficacy of LR cFFP transfused for coagulation factor replacement in dogs. However, owing to the small sample size and high variability of results in the present study, additional research with a larger sample size is required for definitive conclusions on the effects of leukoreduction on coagulation factors in cFFP and to develop treatment guidelines for LR cFFP use in dogs with congenital and acquired coagulopathies.

Porcine reproductive and respiratory syndrome (PRRS), which is caused by the PRRS virus (PRRSV), has resulted in large economic losses for the swine industry. The virus has shown remarkable genetic diversity since its discovery. In our study, we investigated mutation types in the evolution of PRRSV for both in vivo and in vitro passaging of the virus. Sequence alignment analysis demonstrated that the most common hypermutations expressed were A→G/U→C and G→A/C→U. The data provide a new theoretical basis for PRRSV evolution.

OBJECTIVE To evaluate the use of a modified passive leg-raising maneuver (PLRM) to predict fluid responsiveness during experimental induction and correction of hypovolemia in isoflurane-anesthetized pigs. ANIMALS 6 healthy male Landrace pigs. PROCEDURES Pigs were anesthetized with isoflurane, positioned in dorsal recumbency, and instrumented. Following induction of a neuromuscular blockade, pigs were mechanically ventilated throughout 5 sequential experimental stages during which the blood volume was manipulated so that subjects transitioned from normovolemia (baseline) to hypovolemia (blood volume depletion, 20% and 40%), back to normovolemia, and then to hypervolemia. During each stage, hemodynamic variables were measured before and 3 minutes after a PLRM and 1 minute after the pelvic limbs were returned to their original position. The PLRM consisted of raising the pelvic limbs and caudal portion of the abdomen to a 15° angle relative to the horizontal plane. RESULTS Hemodynamic variables did not vary in response to the PLRM when pigs were normovolemic or hypervolemic. When pigs were hypovolemic, the PLRM resulted in a significant increase in cardiac output and decrease in plethysomographic variability index and pulse pressure variation. When the pelvic limbs were returned to their original position, cardiac output and pulse pressure variation rapidly returned to their pre-PLRM values, but the plethysomographic variability index did not. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested a modified PLRM might be useful for identification of hemodynamically unstable animals that are likely to respond to fluid therapy. Further research is necessary to validate the described PLRM for prediction of fluid responsiveness in clinically ill animals.

OBJECTIVE To evaluate the in vitro effect of 20% N-acetylcysteine (NAC) on the viscosity of normal canine bile. ANIMALS Bile samples obtained from 10 adult dogs euthanized for reasons unrelated to biliary disease. PROCEDURES Each sample was centrifuged to remove particulates, then divided into 3 aliquots. One aliquot remained untreated (control). Each of the other aliquots was diluted 1:4 with 20% NAC or sterile water. The viscosity of all samples was measured with a rotational viscometer at 25°C. Viscosity of control samples was measured immediately after centrifugation and at 1 and 24 hours after treatment application to the diluted samples. Viscosity of diluted samples was measured at 1 and 24 hours after treatment application. RESULTS Mean viscosity differed significantly among the 3 groups at both 1 and 24 hours after treatment application. Relative to control samples, the addition of NAC and sterile water decreased the viscosity by approximately 3.35 mPa·s (95% confidence interval [CI], 1.58 to 5.12 mPa·s) and 2.74 mPa·s (95% CI, 1.33 to 4.14 mPa·s), respectively. Mean viscosity of the NAC-treated samples was approximately 0.61 mPa·s (95% CI, 0.21 to 1.01 mPa·s) less than that for the sterile water–treated samples. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that in vitro dilution of canine bile 1:4 with 20% NAC significantly decreased the viscosity of the resulting mixture. Further research is necessary to determine whether NAC is a safe and effective noninvasive treatment for dogs with persistent biliary sludge or gallbladder mucoceles.

OBJECTIVE To determine the relationship between acute volume overload and echocardiographic indices of right ventricular (RV) function and dyssynchrony in healthy dogs. ANIMALS 7 healthy Beagles. PROCEDURES Right heart catheterization and echocardiography were performed in 7 healthy anesthetized Beagles at baseline and after induction of volume overload. Volume overload was induced by IV infusion of lactated Ringer solution (150 mL/kg/h for 90 minutes). Echocardiographic indices of RV function, including peak velocity of systolic tricuspid annular motion, tricuspid annulus plane systolic excursion, fractional area change, RV Tei index, RV longitudinal strain (RVLS), and systolic RV longitudinal strain rate (RVLSR), were obtained by use of speckle tracking echocardiography (STE). In addition, SD of the systolic shortening time of the right ventricle for the 6 segments (RV-SD6) was determined with STE. RESULTS Volume overload significantly increased the RV end-diastolic pressure, compared with the baseline value. Echocardiographic indices of RV function, except for septal RVLSR, were significantly enhanced by volume overload. In contrast, RV-SD6 did not change with volume overload. Although echocardiographic indices of RV function, except for septal RVLSR, were correlated with RV end-diastolic pressure, RV-SD6 was not correlated. CONCLUSIONS AND CLINICAL RELEVANCE Echocardiographic indices of RV function, including RVLS and RVLSR, were affected by acute short-term volume overload. Therefore, results for assessment of RV function by use of STE in dogs with clinical conditions associated with right-sided chronic volume overload, such as tricuspid and pulmonic regurgitation, should be interpreted with caution.

With acrylic external-fixation frames for fracture repair the acrylic columns can be contoured to allow greater versatility in the placement of transfixation pins, thus minimizing damage to the surrounding soft tissue and making mandibular and transarticular fixation easier. However, contouring affects the stiffness and ultimate strength of the construct under axial compression. In this study, polymethylmethacrylate columns 21 mm in diameter angled at 0°, 30°, 45°, 60°, or 90° with clamps were constructed. For each angulation group, pins 3.2-mm long were placed in 6 columns, 2 pins at each end, 1.5 cm from each other, and 6 columns had no pins. Each column was allowed to polymerize for a minimum of 10 min, then was placed in a biomechanical-testing machine, the load cell at the bottom end of the column and the actuator on top, with a preload of 10 to 12 N to prevent slippage. The columns underwent axial loading at a rate of 8 mm/s until catastrophic failure occurred. Data on force and deformation were collected every 0.025 s. Both stiffness and ultimate strength of the column decreased significantly (P < 0.01), up to 77% and 70%, respectively, with each increase of angulation. The columns with pins were significantly less stiff (P < 0.05) than those without pins at angulations of 45° and 60°. However, the columns with pins did not show significant differences in ultimate strength from the columns without pins at any of the angulations. The point of failure was always at the angle of the column, demonstrating that in axial compression the weakest point is not the pin-acrylic interface when pins are eccentrically located within the column.

OBJECTIVE To compare ultracentrifugation, precipitation, and membrane affinity chromatography methods for isolation of extracellular vesicles (EVs) from canine plasma samples and to identify suitable reference genes for incorporation into a quantitative reverse transcription PCR assay of microRNA expression in plasma EVs of healthy dogs. ANIMALS 6 healthy Beagles. PROCEDURES Plasma samples were obtained from each dog, and EVs were isolated from 0.3 mL of these samples via ultracentrifugation, precipitation, and membrane-affinity chromatographic methods. Nanoparticle tracking analysis was performed to determine the concentration and size distribution of EVs isolated by the ultracentrifugation method. Expression levels (cycle threshold values) of 4 microRNAs (let-7a, miR-16, miR-26a, and miR-103) were then compared by means of quantitative reverse transcription PCR assay. Three statistical programs were used to identify the microRNAs most suitable for use as reference genes. RESULTS Results indicated that ultracentrifugation was the most stable of all 3 methods for isolating microRNAs from 0.3 mL of plasma. Nanoparticle tracking revealed that EV samples obtained by the ultracentrifugation method contained a mean ± SD of approximately 1.59 × 10(10) vesicles/mL ± 4.2 × 10(8) vesicles/mL. Of the 4 microRNAs in plasma EVs isolated by ultracentrifugation, miR-103 was the most stable. CONCLUSIONS AND CLINICAL RELEVANCE The ultracentrifugation method has potential as a stable method for isolating EVs from canine plasma samples with a high recovery rate, and miR-103 may provide the most stable reference gene for normalizing microRNA expression data pertaining to plasma EVs isolated by ultracentrifugation.