A fast, sensitive, and specific reverse-transcription (RT) loop-mediated isothermal amplification (RT-LAMP) assay was developed that involved a single tube and a 1-step reaction for detecting infectious bursal disease virus (IBDV). Four specific primers were used for amplification of the VP2 gene of IBDV. The amplified LAMP products were detected by DNA electrophoresis and by direct observation with the naked eye in the presence of SYBR Green I. The sensitivity of RT-LAMP was determined to be 0.01 fg of IBDV viral RNA. This assay for IBDV is more sensitive than the conventional RT-polymerase chain reaction assay, which has a detection limit of 1 ng. The LAMP assay was also assessed for specificity and was found to precisely discriminate between positive and negative test samples. This newly established LAMP assay, combined with RT, is a practical diagnostic tool because IBDV-infected and uninfected clinical samples collected from an experimental farm could be discriminated. Full verification of a sample’s IBDV status was obtained within 40 min of extraction of the viral RNA, which could then be directly added to the RT-LAMP reaction mixture.

Objective: To evaluate influence of electrode position on cardioversion energy (CE; energy delivered in the shock at which cardioversion was achieved) during transvenous electrical cardioversion (TVEC) in horses with atrial fibrillation. Animals: 37 horses with atrial fibrillation (41 cardioversion events). Procedures: Records were reviewed to identify horses that underwent TVEC for treatment of atrial fibrillation. Signalment and CE were recorded. Electrode positions in the right atrium and pulmonary artery were identified on intraoperative radiographs. An orthogonal coordinate space was created, and electrode y- and z-axis coordinates and shadow lengths were determined. Trigonometric modeling was used to estimate x-axis electrode positions that resulted in observed shadows. Postmortem casts of catheterized horses were used to assess electrode paths and anatomic relationships. Model assumptions were tested by use of these and a theoretical data set. Relationships between signalment, electrode position, and CE were assessed via multivariate analysis. Results: Sex and y-axis differences between electrode positions were significant predictors of CE. Population stratification based on examination of residuals improved model strength; populations differed in z-axis variables and in CE. Decreasing distance between electrodes and pulmonary artery electrode positions ventral to the right atrium were associated with increased CE. Agreement between estimated and actual x-axis coordinates was poor. Conclusions and Clinical Relevance: Optimal electrode positioning can reduce the energy requirement for successful TVEC and may eventually support application of TVEC under short-term IV anesthesia and potentially increase chances of treatment response. Further investigation into these relationships is warranted.

Objective—To develop and determine the feasibility of a novel minimally invasive technique for percutaneous catheterization and embolization of the thoracic duct (PCETD) in dogs and to determine thoricic duct TD pressure at rest and during short-term balloon occlusion of the cranial vena cava (CrVC). Animals—Fifteen 7- to 11-month-old healthy mixed-breed dogs. Procedures—Efferent intestinal lymphangiography was performed, and the cisterna chyli was punctured with a trochar needle percutaneously under fluoroscopic guidance. When access was successful, a guide wire was directed into the TD through the needle and a vascular access sheath was advanced over the guide wire. Thoracic duct pressure was measured at rest and during acute balloon occlusion of the CrVC. The TD was then embolized cranial to the diaphragm with a combination of microcoils and cyanoacrylate or ethylene vinyl alcohol. Results—Successful puncture of the cisterna chyli with advancement of a wire into the TD was possible in 9 of 15 dogs, but successful catheterization was possible in only 5 of 9 dogs. Acute balloon occlusion of the CrVC led to a substantial TD pressure increase in 4 of 4 dogs, and embolization of the TD was successful in 4 of 4 dogs. Conclusions and Clinical Relevance—PCETD can successfully be performed in healthy dogs; however, this minimally invasive technique cannot currently be recommended for routine treatment of chylothorax, in part because of the technically demanding nature of the procedure. An increase in jugular venous pressure led to an increase in TD pressure, potentially predisposing some dogs to developing chylothorax.

Objective—To determine expression of cyclooxygenase (COX) genes 1 and 2 (also called prostaglandin-endoperoxide synthases 1 and 2) and stability of housekeeping gene expression during low-flow ischemia and reperfusion in the jejunum of horses. Animals—5 healthy adult horses. Procedures—Horses were anesthetized, and two 30-cm segments of jejunum were surgically exteriorized. Blood flow was maintained at baseline (untreated) values in 1 (control) segment and was decreased to 20% of baseline (low-flow ischemia) for 75 minutes, followed by 75 minutes of reperfusion, in the other (experimental) segment. Biopsy samples were collected from experimental segments at baseline (T0), after 75 minutes of ischemia (T1), and after 75 minutes of reperfusion (T2); samples were collected from control segments at T0 and T2. Horses were euthanized 24 hours after induction of ischemia (T3), and additional samples were collected. Samples were evaluated histologically. Total RNA was extracted; expression of COX genes and stability of 8 housekeeping genes were determined via quantitative real-time PCR assays. Results—COX-1 and COX-2 genes were constitutively expressed in baseline samples. Low-flow ischemia resulted in significant upregulation of COX-2 gene expression at each subsequent time point, compared with baseline values. The most stably expressed reference genes were β-actin and hypoxanthine phosphoribosyltransferase, whereas glyceraldehyde 3-phosphate dehydrogenase and β-2 microglobulin were the least stably expressed. Conclusions and Clinical Relevance—Low-flow ischemia resulted in upregulation of COX-2 gene expression in the jejunum of horses. Housekeeping genes traditionally used as internal standards may not be stable in this tissue during arterial low-flow ischemia and reperfusion.

A useful approach for evaluating antitussive drugs in humans is to determine the sensitivity of the cough reflex to a standard challenge. The purpose of this study was to determine if methods used to induce coughing in humans would be effective when used on awake, untrained, healthy dogs for future application in therapeutic trials involving dogs with spontaneous disease. Methods tested were: mechanically stimulating the trachea by digital compression as well as by vibration from an electric shaver, neck massager, and palm sander (11 dogs), and administering nebulized irritant (3000 micromolar capsaicin), acidic (1 M citric acid), and hypotonic (deionized water) solutions using face masks (4 dogs). The threshold for success was defined as induction of at least 2 moderate or strong coughs in at least 75% of the dogs. None of the methods tested was successful. Digital compression induced soft (n = 2) or moderate (n = 1) coughing in 3 of 11 dogs tested. Nebulization of citric acid induced 1 soft cough in 1 of 4 dogs. It was concluded that coughing cannot be successfully induced in awake, healthy dogs using methods that are successful in humans. Other strategies must be developed so that cough sensitivity can be objectively and non-invasively measured in dogs for clinical research purposes.

Objective: To determine whether therapeutic concentrations of levetiracetam can be achieved in cats and to establish reasonable IV and oral dosing intervals that would not be associated with adverse effects in cats. Animals: 10 healthy purpose-bred cats. Procedures: In a randomized crossover study, levetiracetam (20 mg/kg) was administered orally and IV to each cat. Blood samples were collected 0, 10, 20, and 40 minutes and 1, 1.5, 2, 3, 4, 6, 9, 12, and 24 hours after administration. Plasma levetiracetam concentrations were determined via high-performance liquid chromatography. Results: Mean ± SD peak concentration was 25.54 ± 7.97 μg/mL. The mean y-intercept for IV administration was 37.52 ± 6.79 μg/mL. Half-life (harmonic mean ± pseudo-SD) was 2.95 ± 0.95 hours and 2.86 ± 0.65 hours for oral and IV administration, respectively. Mean volume of distribution at steady state was 0.52 ± 0.09 L/kg, and mean clearance was 2.0 ± 0.60 mL/kg/min. Mean oral bioavailability was 102 ± 39%. Plasma drug concentrations were maintained in the therapeutic range reported for humans (5 to 45 μg/mL) for at least 9 hours after administration in 7 of 10 cats. Only mild, transient hypersalivation was evident in some cats after oral administration. Conclusions and Clinical Relevance: Levetiracetam (20 mg/kg) administered orally or IV to cats every 8 hours should achieve and maintain concentrations within the therapeutic range for humans. Levetiracetam administration has favorable pharmacokinetics for clinical use, was apparently tolerated well, and may be a reasonable alternative antiepileptic drug in cats.

Objective—To evaluate symmetry of the hind limbs in orthopedically normal trotting dogs. Animals—19 orthopedically normal Labrador Retrievers with no history of lameness. Procedures—Retroreflective markers were applied to the hind limb joints, and a 4-camera kinematic system captured positional data at 200 Hz in tandem with force platform data collection while the dogs trotted. Morphometric data were combined with kinematic and force data in an inverse dynamics method to calculate net joint moments and powers at the joints as well as total support moment for each limb. Dogs were identified as right or left dominant when their total support moment was > 10% asymmetric between sides. Results—10 of the 19 dogs were mechanically dominant in the right hind limb as determined by their total support moments. One dog was left dominant, and the remaining 8 were symmetric. Right-dominant dogs had larger net joint moments at the right hip, tarsal, and metatarsophalangeal joints and a smaller moment at the right stifle joint, compared with values for the left hind limb. The 1 left-dominant dog had the exact opposite findings. Hip and stifle joint moments and powers varied between limbs of the right-dominant and left-dominant groups in the timing of their transition from negative to positive, and power amplitudes varied at the hip, tarsal, and metatarsophalangeal joints but not the stifle joint. Conclusions and Clinical Relevance—Sound trotting dogs can have asymmetries in limb and joint mechanics. These natural mechanical asymmetries should be taken into account when considering models to evaluate stresses at joints and when considering surgery for cruciate ligament rupture.

Escherichia coli O157:H7 remains a threat to humans via cattle-derived fecal contamination of food and water. Preharvest intervention strategies represent a means of reducing the pathogen burden before harvest. In this study, the efficacy of a commercially produced type III secreted protein (TTSP) vaccine was evaluated with the use of a commingled experimental calf infection model (30 placebo-treated animals and 30 vaccinates). The calves were vaccinated on days 0, 21, and 42 and infected with 109 colony-forming units (CFU) of E. coli O157 by oral–gastric intubation on day 56. Fecal shedding was monitored daily for 14 d. Serologic assessment revealed a robust immune response to vaccination; the serum titers of antibodies against EspA, Tir, and total TTSPs were significantly higher in the vaccinates than in the placebo-treated animals on days 21, 42, 56, and 70. Significantly less (P = 0.011) of the challenge organism was shed by the vaccinates than by the placebo-treated animals on days 3 to 10. Peak shedding occurred in both groups on days 3 to 6; during this period the vaccinates showed a mean log reduction of 1.4 (P = 0.002) and a mitigated fraction of 51%. The number of animals shedding was significantly lower among the vaccinates compared with the placebo group on days 3 to 6 (P ≤ 0.05), with a mean prevented fraction of 21%. No differences in the duration of shedding were observed. Owing to the low challenge shedding in both groups on days 11 to 14 (mean CFU/g < 10; median = 0), no significant differences were observed. These data indicate that TTSP vaccination had protective effects through significant reductions in the number of animals shedding and the number of challenge organisms shed per animal and provides evidence that TTSP vaccination is an effective preharvest intervention strategy against E. coli O157.

Objective—To determine whether water content in a canned food diet induces decreases in voluntary energy intake (EI) or body weight (BW) in cats fed ad libitum. Animals—16 sexually intact male domestic shorthair cats. Procedures—Maintenance EI was determined for 2 months in 10 weight-stable cats consuming a control diet (typical colony diet). Cats were allocated into 2 groups of equal BW and fed a canned diet (with-water [WW] diet) or a freeze-dried version of the canned diet (low-water [LW] diet) twice daily. Diets were identical in nutrient profile on a dry-matter basis. Each dietary treatment period of the crossover experiment lasted 3 weeks, with a 3-week washout period between diets. Body composition measurements were determined by use of deuterium oxide at the end of each dietary treatment. Daily food intake was measured for determination of dry-matter intake and EI. Six other cats were used in preference tests for the 3 diets. Results—EI was significantly decreased for the WW diet (mean ± SD, 1,053.0 ± 274.9 kJ/d), compared with EI for the LW diet (1,413.8 ± 345.8 kJ/d). Cats had a significant decrease in BW during consumption of the WW diet. Body composition was unaltered by diet. In short-term preference tests, cats ate significantly more of the WW than the LW diet. Conclusions and Clinical Relevance—Bulk water in the WW diet stimulated decreases in EI and BW in cats. The impact of water content on energy density and food consumption may help promote weight loss in cats.

Objective: To determine the impact of a free-choice diet on nutritional intake and body condition of feral horses. Animals: Cadavers of 41 feral horses from 5 Australian locations. Procedures: Body condition score (BCS) was determined (scale of 1 to 9), and the stomach was removed from horses during postmortem examination. Stomach contents were analyzed for nutritional variables and macroelement and microelement concentrations. Data were compared among the locations and also compared with recommended daily intakes for horses. Results: Mean BCS varied by location; all horses were judged to be moderately thin. The BCS for males was 1 to 3 points higher than that of females. Amount of protein in the stomach contents varied from 4.3% to 14.9% and was significantly associated with BCS. Amounts of water-soluble carbohydrate and ethanol-soluble carbohydrate in stomach contents of feral horses from all 5 locations were higher than those expected for horses eating high-quality forage. Some macroelement and microelement concentrations were grossly excessive, whereas others were grossly deficient. There was no evidence of ill health among the horses. Conclusions and Clinical Relevance: Results suggested that the diet for several populations of feral horses in Australia appeared less than optimal. However, neither low BCS nor trace mineral deficiency appeared to affect survival of the horses. Additional studies on food sources in these regions, including analysis of water-soluble carbohydrate, ethanol-soluble carbohydrate, and mineral concentrations, are warranted to determine the provenance of such rich sources of nutrients. Determination of the optimal diet for horses may need revision.