Objective—To determine the correlation between activity as measured by an accelerometer and videographic measurements of movement and mobility in healthy dogs.
Animals—4 healthy dogs.
Procedures—After determination that accelerometers had good agreement, 5 identical accelerometers were used simultaneously to test their output at 8 locations (rotated among collar, vest, and forelimb stocking locations) on each dog. Movement and mobility for each dog were recorded continuously with a computerized videography system for 7-hour ses-sions on 4 consecutive days. Accelerometer values were combined into 439 fifteen-minute intervals and compared with 3 videographic measurements of movement and mobility (distance traveled, time spent walking > 20 cm/s, and time spent changing position by > 12% of 2-dimensional surface area during 1.5 seconds).
Results—96% of values compared between the most discordant pair of accelerometers were within 2 SDs of the mean value from all 5 accelerometers. All mounting locations provided acceptable correlation with videographic measurements of movement and mobility, and the ventral portion of the collar was determined to be the most convenient location.
Conclusions and Clinical Relevance—Use of an accelerometer was adequate for at-home activity monitoring, an important end point in clinical trials of treatment for chronic disease, and provided information about daily activity that is unattainable by other methods.
Objective—To develop a clinically applicable assay for detection of serum anti-neutrophil antibodies in dogs.
Sample Population—Serum samples of 20 healthy dogs and 20 sick dogs.
Procedures—An indirect immunofluorescence assay was developed in which canine serum was incubated with paraformaldehyde-fixed neutrophils and subsequently incubated with fluorescein-conjugated rabbit anti-dog IgG. Neutrophil median fluorescence intensity and the percentage of neutrophils with an increase in fluorescence intensity were determined by use of a flow cytometer.
Results—Neutrophils incubated with serum from healthy and sick dogs had a normally distributed curve when displayed as a histogram. Alloantibodies or immune complexes that significantly affected test results were not detected. Hyperglobulinemia did not appear to affect test results. The neutrophil donor did not significantly affect test results. With 1 exception, results for the sick dogs did not differ appreciably from those for healthy dogs. Serum from a dog with steroid-responsive neutropenia had a greater neutrophil fluorescence value and percentage of neutrophils with an increase in fluorescence intensity, compared with either healthy or sick dogs.
Conclusions and Clinical Relevance—The indirect immunofluorescence test gave consistent results for healthy and sick dogs and detected anti-neutrophil antibodies in a dog with steroid-responsive neutropenia. Definitive evaluation of the test will be dependent on evaluation of persistently neutropenic dogs and correlation of test results with a response to immunosuppressive therapy.
Objective—To assess agreement between a commercially available Geiger-Meuller (GM) survey meter and millirem tissue-equivalent (TE) meter for measuring radioactivity in cats treated with sodium iodine I 131 (131I).
Animals—15 cats with hyperthyroidism and undergoing 131I treatment.
Procedures—Duplicate measurements were obtained at a distance of 30 cm from the thyroid region of each cat's neck by 2 observers who used both meters on day 3 or 5 after131I administration. Comparisons of measurements between meters and observers were made, with limits of agreement defined as the mean difference ± 2 SDs of the differences.
Results—For observer 1, the mean of the differences in the 2 meters' measurements in all cats was 0.012 mSv/h (SD, 0.011 mSv/h). For observer 2, the mean of the differences in measurements was 0.012 mSv/h (SD, 0.010 mSv/h). For the GM meter, the mean of the differences of the 2 observers for all cats was 0.003 mSv/h (SD, 0.011 mSv/h). For the TE meter, the mean of the differences of the 2 observers for all cats was 0.003 mSv/h (SD, 0.007 mSv/h).
Conclusions and Clinical Relevance—Results indicate that there was considerable agreement between meters and observers in measurements of radioactivity in cats treated with 131I. Measurements obtained by use of the GM meter may be approximately 0.01 mSv/h less than or 0.03 mSv/h higher than those obtained with the TE meter. If this range is acceptable for an institution's release criteria, the 2 meters should be considered interchangeable and acceptable for clinical use.
Objective—To evaluate the use of urinary biomarkers to assess exposure of cats to environmental tobacco smoke (ETS).
Animals—61 healthy client-owned cats (19 from households in which smoking was reported and 42 from households in which there was no smoking).
Procedures—Urine samples were obtained from each cat and assayed for total nicotine (nicotine plus nicotine glucuronide) and total cotinine (cotinine plus cotinine glucuronide) content by use of gas chromatography-mass spectrometry. In addition, total urinary content of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a major metabolite of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, was measured by use of gas chromatography with nitrosamine-selective detection.
Results—Cats from households in which smoking was reported had significantly higher concentrations of total nicotine (70.4 ng/mL), total cotinine (8.53 ng/mL), and total NNAL (0.0562 pmol/mL) in urine, compared with concentrations for cats that lived in households in which there was no smoking (4.89 ng/mL, 0.74 ng/mL, and 0.0182 pmol/mL, respectively).
Conclusions and Clinical Relevance—Analysis of these data provided biochemical evidence of exposure to ETS and uptake of tobacco-specific carcinogens by cats that live in households with smokers. Biomarkers could facilitate investigation of the health effects of ETS in cats and other species.
Objective—To develop quantitative PCR (qPCR) assays with allele-specific primers to provide a rapid and accurate diagnostic and screening test for the 3 mutations identified as causes of gangliosidoses in domestic cats.
Sample Population—DNA samples obtained from archived feline blood samples submitted for GM1 and GM2 testing.
Procedures—A qPCR assay was developed for each mutation to monitor the efficiency of PCR amplification. Results were determined on the basis of the fluorescent intensity of DNA staining.
Results—Samples from 60 cats were screened by use of the 3 qPCR assays. Of these, 59 qPCR results agreed with the sequence-derived genotypes. The phenotype (affected) for the other cat agreed with results for the qPCR assay, which indicated that interpretation of the sequence-based result was incorrect.
Conclusions and Clinical Relevance—The qPCR assays offer a sensitive, rapid, and reproducible technique for allelic discrimination without the need for complicated processing steps, such as hybridization or sequencing, after PCR procedures. These assays may prove beneficial for a rapid diagnosis of gangliosidoses in cats and could also provide a means for reliable large-scale screening for the carrier state, thereby accelerating the eradication of these debilitating diseases from feline populations.
Objective—To estimate the sensitivity (Se) and specificity (Sp) for an enhanced direct-fecal PCR procedure, bacterial culture of feces (BCF), and a serum ELISA for detecting Mycobacterium avium subsp paratuberculosis (MAP) infection in adult dairy cattle.
Sample Population—Fecal and serum samples were collected from 669 adult cattle randomly selected from a 4,000-cow dairy herd known to contain animals infected with MAP.
Procedures—Serum samples were evaluated for MAP-specific antibodies via ELISA. Fecal samples were evaluated by BCF and enhanced PCR methods (both gel-based [GB]-PCR and quantitative real-time [qRT]-PCR assays). Fecal samples also were pooled (5:1) and then subjected to GB-PCR assay. Bayesian statistical methods were used to estimate Se and Sp for each diagnostic test without knowledge concerning true MAP infection status.
Results—Adjusting for Se conditional dependence between serum ELISA and BCR, overall Se and Sp were estimated at 33.7% and 95.9%, 51.3% and 99.0%, and 32.2% and 100% for serum ELISA, qRT-PCR, and BCF, respectively.The GB-PCR assay yielded positive results for 38.3% of the pools known to contain feces from at least 1 cow that had positive GBPCR results.
Conclusions and Clinical Relevance—Estimated Se values for the serum ELISA and BCF were slightly lower than those reported elsewhere. The enhanced qRT-PCR method offered relative improvements in Se of 52% and 59% over serum ELISA and microbial culture, respectively. Pooling of fecal samples and testing with the GB-PCR assay are not recommended. Additional studies with qRT-PCR and fecal pools are required.
Objective—To analyze a centrifugation-based, point-of-care device that concentrates canine platelets and bone marrow–derived cells.
Animals—19 adult sexually intact dogs.
Procedures—Anticoagulated peripheral blood (60 mL) and 60 mL of anticoagulated bone marrow aspirate (BMA) were concentrated by centrifugation with the centrifugation-based, point-of-care device to form a platelet and a bone marrow concentrate (BMC) from 11 dogs. Blood samples were analyzed on the basis of hemograms, platelet count, and PCV. The BMA and BMC were analyzed to determine PCV, total nucleated cell count, RBC count, and differential cell counts. The BMC stromal cells were cultured in an osteoinductive medium. Eight additional dogs were used to compare the BMC yield with that in which heparin was infused into the bone marrow before aspiration.
Results—The centrifugation-based, point-of-care device concentrated platelets by 6-fold over baseline (median recovery, 63.1%) with a median of 1,336 × 103 platelets/μL in the 7-mL concentrate. The nucleated cells in BMCs increased 7-fold (median recovery, 42.9%) with a median of 720 × 103 cells/μL in the 4-mL concentrate. The myeloid nucleated cells and mononuclear cells increased significantly in BMCs with a significant decrease in PCV, compared with that of BMAs. Stromal cell cultures expressed an osteoblastic phenotype in culture. Infusion of heparin into the bone marrow eliminated clot formation and created less variation in the yield (median recovery, 61.9%).
Conclusions and Clinical Relevance—Bone marrow–derived cell and platelet-rich concentrates may form bone if delivered in an engineered graft, thus decreasing the need for cancellous bone grafts.
Objective—To compare 2 methods of quantitating chondrocyte viability and to determine chondrocyte response to thermal injury over time.
Sample Population—108 stifle joints from 54 adult rats.
Procedures—Cartilage from the distal aspect of the femur was treated ex vivo with radiofrequency energy at a probe setting that would result in immediate partial-thickness chondrocyte death; untreated sections served as controls. Explants were cultured, and cell viability was compared by use of lactate dehydrogenase (LDH) histochemical staining and calcein AM and ethidium homodimer-1 confocal laser microscopy (CLM) cell viability staining. Terminal deoxynucleotidyl transferase–mediated X-dUTP nick end labeling (TUNEL) was used to detect apoptosis. All labeling studies were performed 0, 1, 3, 7, 14, and 21 days after treatment.
Results—In the treated tissues, a greater percentage of viable cells were found with CLM, compared with LDH staining. This result contrasted that of control tissues in which LDH staining indicated a greater percentage of live cells than CLM. The greatest number of TUNEL-positive chondrocytes was present at day 3, declining at later time intervals.
Conclusions and Clinical Relevance—CLM and LDH histochemistry techniques yield different absolute numbers of live and dead cells, resulting in differing percentages of live or dead cells with each technique. These differences may be related to the enzymes responsible for activation in each technique and the susceptibility of these enzymes to thermal injury. Results of TUNEL indicate that apoptosis contributes to chondrocyte death after thermal injury, with a peak signal identified 3 days after insult.
Objective—To determine an optimal dose of carbon 13 (13C)-labeled aminopyrine for use in a 13C-aminopyrine demethylation blood test in healthy dogs.
Animals—9 adult dogs.
Procedures—Food was withheld from each dog for 12 hours. A 2-mL baseline blood sample was obtained from each dog and placed into an evacuated tube containing sodium heparin. Carbon 13-labeled aminopyrine was administered IV at doses of 1, 2, 5, or 10 mg/kg. Additional blood samples (2 mL) were obtained and placed into evacuated tubes containing sodium heparin 30, 45, 60, and 75 minutes after 13C-aminopyrine administration. Hydrochloric acid was used to extract CO2 from blood samples. The extracted gas was analyzed by fractional mass spectrometry to determine the percentage dose of 13C administered as 13C-aminopyrine and recovered in extracted gas (PCD).
Results—Gross evidence of clinical adverse effects was not detected in any dog after administration of 13C-aminopyrine. The mean coefficient of variation (CV) for PCD was significantly lower than the mean CV for the summation of PCD values up to a given sampling time (CUMPCD). Mean PCD values among the 4 doses for each sample time were not significantly different. Administration of 13C-aminopyrine at a dose of 2 mg/kg resulted in the lowest interindividual variability.
Conclusions and Clinical Relevance—The PCD is superior to CUMPCD for the quantification of aminopyrine demethylation. Administration of 13C-13C-aminopyrine at a dose of 2 mg/kg is appropriate for use in the 13C-aminopyrine demethylation blood test in healthy dogs.
Objective—To determine the effects of indwelling nasogastric intubation on the gastric emptying rate of liquid in horses.
Animals—6 healthy horses.
Procedures—Horses were assigned to treatment and control groups in a prospective randomized crossover study with a washout period of at least 4 weeks between trials. Acetaminophen (20 mg/kg) diluted in 1 L of distilled water was administered via nasogastric tube at time points of 0, 12, 30, 48, and 72 hours to evaluate the liquid-phase gastric emptying rate. In control horses, nasogastric tubes were removed after administration of acetaminophen. In horses receiving treatment, the tube was left indwelling and maintained for 72 hours. A 10-mL sample of blood was collected from a jugular vein immediately before and 20, 40, 60, 80, 100, 120, and 180 minutes after acetaminophen administration. Serum acetaminophen concentrations were measured by use of a colorimetric method.
Results—Peak serum acetaminophen concentration was significantly higher in the control group (38.11 μg/mL) than in the treatment group (29.09 μg/mL), and the time required to reach peak serum acetaminophen concentration was significantly shorter in the control group (22.79 minutes) than in the treatment group (35.95 minutes).
Conclusions and Clinical Relevance—Results indicated that indwelling nasogastric intubation has a delaying effect on the gastric emptying rate of liquids. Veterinarians should consider the potential for delayed gastric emptying when placing and maintaining an indwelling nasogastric tube for an extended period of time after surgery. Repeated nasogastric intubation may be better than maintenance of an indwelling tube in horses with ileus.