peer reviewed | The purpose of this study was to assess the impact of freezing and thawing on MR images of equine feet examined ex vivo. Nine equine cadaver digits were first imaged at room temperature (T0). Among the 9 digits, 3 (group 1) were imaged in a 3 Tesla MR system after one and after 2 freezing-thawing cycles. Digits of group 1 were thawed in a cold room at 4°C for 36h. Three other digits (group 2) were imaged after one freezing-thawing cycle. Digits of group 2 were thawed in a cold room at 4°C and then rescanned after 24h at room temperature. The last 3 digits (group 3) were scanned after one freezing-thawing cycle. Digits of group 3 were thawed at room temperature for 24h. Sequences used were Spin Echo (SE) T1, Turbo Spin Echo (TSE) T2 and proton density (PD), Short Tau Inversion Recovery (STIR), Double Echo Steady State (DESS), 3D Gradient Echo (GE) T1 and 2D GE T2*. Images obtained on the fresh limbs at room temperature were subjectively compared side by side to images obtained at the different freezing-thawing cycles. A quantitative analysis to assess signal change between examinations was realized by measuring signal to noise ratio (SNR). Visibility and margination of the anatomical structures of the foot and overall image quality were subjectively considered unchanged except for the hoof where the lamina was considered less visible distally after freezing and thawing in the GE T2* and in TSE T2 and PD sequences. Quantitative analysis demonstrated SNR changes in the bone marrow only in the distal phalanx in the SE T1 sequence when the feet were thawed at room temperature. When the feet were thawed in a cold room at 4°C, bone marrow SNR changes were present in the SE T1, GE T1 and TSE PD sequences. Signal changes were significant in the synovial recess when the thawing process was made at 4°C and not when the thawing process was at ambient temperature. The soft tissue structures and the hoof capsule showed significant changes with an increase of SNR, except in STIR, after freezing and thawing at 4°C and at room temperature. SNR changes in the soft tissues were mainly present in GE sequences.

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 changes in the cortical bone of the proximal phalanx of the fore-limbs of Thoroughbreds in response to training. Animals—Twenty-seven 2-year-old Thoroughbreds (20 females, 2 males, and 5 geldings). Procedures—Horses were principally in training for races in a straight line and in a clockwise direction. Lateromedial and dorsopalmar radiographic views of each metacarpophalangeal joint were obtained before the horses started training and 1 year after starting exercise and racing. Width of the dorsal, palmar, lateral, and medial cortex and the width and thickness of the medulla were measured. Ratios (rather than absolute values) were used to remove the effect of differences in bone size among horses. Results—10 horses were lost from the study. Radiographs were obtained for 17 horses 1 year after starting training (9 horses raced in a clockwise direction, and 8 raced in clockwise and counterclockwise directions). There was no difference between the cortical bone in the right and left forelimbs at the start of the study. After training for 1 year, the palmar cortex in the right forelimb was significantly thicker than that in the left forelimb. Conclusions and Clinical Relevance—The strain patterns, biomechanics of rapid exercise, and type of training most probably determined differences in the adaptive responses of the proximal phalanx. The data reported here can be used in the evaluation of weight-bearing distribution along the proximal phalanx and evaluation of the relationship between exercise and bone remodelling of the proximal phalanx.

Objective—To compare results of body condition scoring by use of a 9-point scale with body composition determined by dual-energy x-ray absorptiometry (DEXA) in indoor-confined neutered domestic shorthair (DSH) pet cats. Animals—72 indoor-confined, adult neutered DSH pet cats (38 females and 34 males). Procedures—All cats underwent a physical examination including assessment of body weight (BW), body condition score (BCS; 1 = emaciated, 5 = ideal, and 9 = grossly obese), and girth. Urinalysis, CBC, and serum biochemical analysis were also performed. After the cats were confirmed healthy, they were anesthetized for body composition measurement via DEXA. Lean body mass, fat mass, and percentage body fat (%BF) were then evaluated. Results—The correlation between %BF and BCS (r = 0.87) was superior to the correlations between %BFand BW (r = 0.74) and between %BF and girth (r = 0.78). Values for %BF differed significantly between all pairs of BCSs except BCSs 8 and 9. Within a BCS, the %BF was similar for male and female cats. The mean %BF for cats with a BCS of 5 was 32, which exceeded the upper reference limit of %BF generally considered ideal (30). Conclusions and Clinical Relevance—The 9-point BCS scale appears useful for assessing %BF in DSH pet cats. Nevertheless, study findings could indicate a need for redefining the ideal BCS for inactive neutered cats to include a BCS of 4.

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 investigate the density of the primary epidermal lamellae (PEL) around the solar circumference of the forefeet of near-term fetal feral and nonferal (ie, domesticated) horses. Sample—Left forefeet from near-term Australian feral (n = 14) and domesticated (4) horse fetuses. Procedures—Near-term feral horse fetuses were obtained from culled mares within 10 minutes of death; fetuses that had died in utero 2 weeks prior to anticipated birth date and were delivered from live Thoroughbred mares were also obtained. Following disarticulation at the carpus, the left forefoot of each fetus was frozen during dissection and data collection. In a standard section of each hoof, the stratum internum PEL density was calculated at the midline center (12 o'clock) and the medial and lateral break-over points (11 and 1 o'clock), toe quarters (10 and 2 o'clock), and quarters (4 and 6 o'clock). Values for matching lateral and medial zones were averaged and expressed as 1 density. Density differences at the 4 locations between the feral and domesticated horse feet were assessed by use of imaging software analysis. Results—In fetal domesticated horse feet, PEL density did not differ among the 4 locations. In fetal feral horse feet, PEL density differed significantly among locations, with a pattern of gradual reduction from the dorsal to the palmar aspect of the foot. The PEL density distribution differed significantly between fetal domesticated and feral horse feet. Conclusions and Clinical Relevance—Results indicated that PEL density distribution differs between fetal feral and domesticated horse feet, suggestive of an adaptation of feral horses to environment challenges.

Objective--To evaluate 5 commercially available ELISAs for determination of leptin concentrations in serum samples from dogs. Sample Population--Serum samples from overweight-obese and thin–ideal weight clientowned dogs. Procedures--Serum samples with high and low leptin concentrations (n = 7 samples each) were used for validation of the assays. Intra- and interassay precision, linearity under dilution, spiking recovery, and limit of quantification were determined. In addition, leptin concentrations in thin–ideal weight (n = 8) and overweight-obese (37) dogs were quantified. Results--Use of 2 of the 5 ELISAs (A and B) revealed reactivity with canine leptin. Intra-and interassay coefficients of variation were < 6.1% and 76%, respectively, for assay A and 14.0% and 13.7%, respectively, for assay B. In assays A and B, dilutions of canine serum pools were used to determine linear regression equations. Recoveries were 77% to 101% for assay A and 67% to 125% for assay B. Significant differences in leptin concentrations between thin–ideal weight and overweight-obese dogs were detected only when analyzed with assay A. Conclusions and Clinical Relevance--Among 5 leptin ELISAs evaluated, a canine-specific leptin ELISA had adequate precision, linearity, and ability to discriminate between high and low leptin concentrations corresponding to overweight-obese and thin–ideal weight dogs, respectively.

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 noninvasively evaluate physiologic postprandial adaptations of the heart in snakes. Animals: 6 juvenile Paraguay anacondas (Eunectes notaeus). Procedures: The heart of each anaconda was echocardiographically evaluated after food was withheld for 28 days as well as 3 and 10 days after feeding. Physical measurements included body length, weight, and circumference at the level of the heart. Echocardiographic measurements included heart rate and 2-D total and internal ventricular area. From these measurements, total ventricular volume as well as the myocardial area as a surrogate of myocardial mass was calculated. Results: No significant changes in body length, weight, and circumference were found. Significant increases in heart rate (from 45 to 58 beats/min), total ventricular volume (from 4.63 to 5.54 mL), and myocardial area (from 0.7 to 0.81 cm2) were detected 10 days after feeding, compared with results obtained prior to feeding after food had been withheld for 28 days. No pericardial effusion was detected at any time point. Conclusions and Clinical Relevance: Echocardiographic evaluation of the heart of anacondas was performed, and feeding resulted in concentric cardiac hypertrophy. Physiologic fluctuation of cardiac dimensions should be considered when cardiac imaging is performed in snakes.

Objective—To evaluate the diagnostic value of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentrations in Doberman Pinschers in various stages of dilated cardiomyopathy (DCM). Animals—328 Doberman Pinschers. Procedures—Staging of DCM was determined via analysis of results of physical examinations, 24-hour ambulatory ECG (Holter) recordings, and echocardiographic evaluations. Plasma samples for NT-proBNP assays were obtained at each examination. Concentrations of NT-proBNP were measured in 337 samples obtained from 196 healthy Doberman Pinschers (control dogs) and in 195 samples obtained from 132 Doberman Pinschers in various stages of DCM. These included dogs that had ventricular premature contractions (VPCs; 79 samples), echocardiographic changes (23 samples), or both (51 samples); 16 samples were from dogs with overt DCM, and 26 were from dogs that were considered normal during initial examination but developed DCM within 1.5 years after this assessment. Receiver operating characteristic curves were analyzed to determine sensitivity and specificity of NT-proBNP concentrations for detection of DCM. Results—NT-proBNP concentrations in dogs that had or developed DCM were significantly higher than those of control dogs. Sensitivity and specificity of NT-proBNP concentrations (cutoff value, > 400 pmol/L) to detect all stages of DCM were 81.1 % and 75.0%, respectively; sensitivity was 90.0% and specificity was 75.0% to predict echocardiographic changes. Specificity to detect echocardiographic changes was 90.4% at a cutoff value of 550 pmol/L. Conclusions and Clinical Relevance—Plasma concentrations of NT-proBNP were increased in dogs with DCM and in apparently healthy dogs that developed DCM within 1.5 years after samples were obtained, compared with concentrations in control dogs.