Validation of Ultrasonography for Assessment of Gastric Emptying Time in Healthy Cats by Radionuclide Scintigraphy.

BACKGROUND: The prevalence of gastric emptying (GE) disorders in cats is unknown due to lack of clinically applicable diagnostic tests. OBJECTIVES: The principal aim of this study was to assess correlation between scintigraphic and ultrasonographic measurements of GE time (GET) in healthy cats. Additionally, variability of ultrasonographic GET, and correlation between scintigraphy and ultrasonographic parameters of gastric motility were evaluated. ANIMALS: Eight healthy domestic shorthair cats. METHODS: Prospective study. Scintigraphic GET was determined using a solid test meal containing 4 mCi 99m Tc-mebrofenin. Each cat had 3 separate ultrasonographic assessments of GE, performed independent of scintigraphic assessment, after solid test meal consumption. The motility index (MI) of antral contractions was plotted against time and time for each fraction of the area under the MI curve determined. Ultrasonographic GET and MI were correlated to scintigraphic GET. RESULTS: Scintigraphic GET (mean ± SD) for 25, 50, and 75% GE was 103 ± 32 minutes, 196 ± 45 minutes, and 288 ± 62 minutes, whereas sonographic GET for 25, 50, and 75% GE was 106 ± 13 minutes, 203 ± 19 minutes, and 305 ± 27 minutes. There was good correlation between scintigraphic and sonographic GET (r = 0.72-0.82) at 45-90% fractional GE and between scintigraphic GET and time of corresponding MI curve fraction (r = 0.78-0.86) at 40-90% fraction of the MI curve. There was moderate intraindividual variability for sonographic GET and MI curve fraction times as well as significant variation among individuals. CONCLUSIONS AND CLINICAL IMPORTANCE: Ultrasonography is a valid alternative to scintigraphy for assessment of solid-phase GE and allows assessment of postprandial gastric motility in healthy cats.

Effect of endoscopic forceps on quality of duodenal mucosal biopsy in healthy dogs.

BACKGROUND: Limited data exist regarding influence of endoscopic forceps on duodenal mucosal biopsy quality and adequacy for histologic examination/assessment in dogs. HYPOTHESIS/OBJECTIVES: Hypothesizing that larger forceps would procure superior specimens, we evaluated effect of 6 disposable forceps on duodenal biopsy weight, depth, crush artifact, and adequacy for histologic examination/assessment. ANIMALS: Seventeen healthy adult dogs. METHODS: Prospective study. Two operators each obtained 4 duodenal specimens from each dog with each forceps. Lightest sample discarded. One pathologist evaluated blindly other 3 specimens. A total of 612 specimens evaluated. Results analyzed by one-way ANOVA of forceps effects with dog as blocking factor. Posthoc pairwise comparisons examined with Tukey's test when indicated. RESULTS: Biopsies performed with large capacity forceps heavier (10.56 ± 0.90 and 11.6 ± 0.62 mg (mean ± SD) versus 5.55 ± 0.53 to 8.61 ± 0.49; P < .0001) and adequacy for histologic examination/assessment superior to standard oval and 'pediatric' (scores 2.52 ± 0.41 and 2.58 ± 0.37 versus 2.08 ± 0.33 and 2.14 ± 0.29; P < .0001). No statistically significant difference in depth scores. Large capacity forceps with spike associated with less crush artifact than all smaller forceps (scores 1.19 ± 0.16 versus 1.38 ± 0.21 to 1.52 ± 0.21; P < .0001). In same size forceps, presence of spike had no effect on crush artifact and adequacy for histologic examination/assessment (P < .0001). CONCLUSIONS AND CLINICAL IMPORTANCE: Large capacity forceps are superior, providing higher quality and greater numbers of samples achieving adequacy for histologic examination/assessment. Choice of endoscopic biopsy forceps for duodenal samples influences sample volume and diagnostic utility.

Evaluation of a wireless ambulatory capsule (SmartPill®) to measure gastrointestinal tract pH, luminal pressure and temperature, and transit time in ponies.

REASONS FOR PERFORMING STUDY: This study investigated the use of a wireless ambulatory capsule (WAC; SmartPill(®) pH.p GI Monitoring System) to determine WAC-gastric emptying time (GET) in ponies. OBJECTIVES: To measure WAC-GET and compare it to those findings with GET assessed by nuclear scintigraphy (S-GET). HYPOTHESIS: WAC-GET will be slower than S-GET, but will be significantly correlated. METHODS: Seven healthy adult mixed-breed pony mares were used in this study. Feed was withheld for 12 h prior to the WAC administration. After administration, a complete-feed diet was fed to allow the WAC to pass into the stomach. Luminal pH, temperature and pressure were collected by a modified receiver secured to the pony. Once the pH reached a value of ≥ 8.0, it was determined that gastric emptying had occurred, and ponies were fed grass hay. After 5 days, data were downloaded and analysed using proprietary software. During the second period of the study, after at least 2 weeks, 4 of the ponies underwent a standard S-GET test. RESULTS: The WAC was successfully administered, and data were collected from all ponies. The mean percentage of data packets collected by the receiver was 84.9 ± 3.51% (range 66.8-95.1%). Mean WAC-GET was 7.38 h (range 0.15-46.65 h). Mean gastric pH was 4.75 (range 2.07-6.99). Mean small intestinal transit time was 4.6 h. The mean pH for the small intestine was 8.0. The mean S-GET time (in hours) when 10% of the radioactive feed is present in the stomach (T-90%) was 2.3 h. The S-GET did not correlate significantly with the WAC-GET. CONCLUSIONS AND POTENTIAL RELEVANCE: The WAC was safely administered to ponies, and data were collected using a modified receiver. The WAC-GET varied considerably between ponies, but was ≤ 3 h in 5 of the 6 ponies. The WAC used in this study provided a noninvasive technique that produced novel information about the pony gastrointestinal tract, but owing to the substantial variability in GET values and long transit time it may not be a reliable clinical tool at this time.

Characterization of Anaplasma phagocytophilum major surface protein 5 and the extent of its cross-reactivity with A. marginale.

Major surface protein 5 (Msp5) of Anaplasma marginale is highly conserved in the genus Anaplasma and the antigen used in a commercially available competitive enzyme-linked immunosorbent assay (cELISA) for serologic identification of cattle with anaplasmosis. This study analyzes the degrees of conservation of Msp5 among various isolates of Anaplasma phagocytophilum and the extent of serologic cross-reactivity between recombinant Msp5 (rMsp5) of Anaplasma marginale and A. phagocytophilum. The msp5 genes from various isolates of A. phagocytophilum were sequenced and compared. rMsp5 proteins of A. phagocytophilum and A. marginale were used separately in an indirect ELISA to detect cross-reactivity in serum samples from humans and dogs infected with A. phagocytophilum and cattle infected with A. marginale. Serum samples were also tested with a commercially available competitive ELISA that uses monoclonal antibody ANAF16C1. There were 100% sequence identities in the msp5 genes among all of the A. phagocytophilum isolates from the United States and a horse isolate from Sweden. Sheep isolates from Norway and dog isolates from Sweden were 99% identical to one another but differed in 17 base pairs from the United States isolates and the horse isolate. Serologic cross-reactivity was identified when serum samples from cattle infected with A. marginale were reacted with rMsp5 of A. phagocytophilum and when serum samples from humans and dogs infected with A. phagocytophilum were reacted with rMsp5 of A. marginale in an indirect-ELISA format. Serum samples from dogs or humans infected with A. phagocytophilum did not cross-react with rMsp5 of A. marginale when tested with the commercially available cELISA. These results suggest that rMsp5 of A. phagocytophilum is highly conserved among United States and European isolates and that serologic distinction between A. phagocytophilum and A. marginale infections cannot be accomplished if rMsp5 from either organism is used in an indirect ELISA.

Mast cell proliferation and alterations in bFGF amount and localization are involved in the response of muscle to dystrophin deficiency in hypertrophic feline dystrophy.

To test the hypothesis that basic fibroblast growth factor and mast cells play a key role in the phenotypic differences between human dystrophinopathies and hypertrophic feline muscular dystrophy, serial sections of dystrophin-deficient, carrier and normal cat muscle biopsy specimens were examined. They were stained immunohistochemically for dystrophin and different markers of differentiation such as desmin, vimentin and utrophin. Basic fibroblast growth factor was increased in the myofibers of dystrophic cats compared to normal controls and carriers. An association of basic fibroblast growth factor with fiber regeneration and necrosis was shown. The amount of mast cells was markedly increased in muscle tissue of dystrophic cats with a clear predominance of tryptase-positive cells present in large amounts in the endomysium. Mast cells, like basic fibroblast growth factor, were concentrated in areas of muscle fiber regeneration and necrosis. Our data concerning basic fibroblast growth factor and mast cells are consistent with a highly abnormal cellular environment in feline dystrophic muscle with very high levels of basic fibroblast growth factor which is likely modulated by mast cells.

Effects of long-term phenobarbital treatment on the thyroid and adrenal axis and adrenal function tests in dogs.

Phenobarbital can interfere with the thyroid axis in human beings and rats by accelerating hepatic thyroxine metabolism because of enzyme induction. In human beings, it also can interfere with the low-dose dexamethasone suppression test (LDDST) used to assess adrenal function by accelerating dexamethasone metabolism. This effect can cause a lack of suppression of pituitary ACTH and subsequent adrenal cortisol release after dexamethasone administration. The effects of phenobarbital on the thyroid axis, the adrenal axis, and adrenal function tests were prospectively investigated in 12 normal, adult dogs. Phenobarbital was administered at 5 mg per kilogram of body weight (range, 4.8-6.6 mg/kg) PO q12h for 29 weeks, resulting in therapeutic serum concentrations (20-40 microg/mL). Serum total thyroxine (TT4), free thyroxine (FT4) by equilibrium dialysis, total triiodothyronine (TT3), thyrotropin (TSH), and cholesterol were determined before and during phenobarbital treatment. LDDST, ACTH stimulation tests, and ultrasonographic evaluation of the adrenal glands were performed before and during treatment. TT4 and FT4 decreased significantly (P < or = .05), TT3 had minimal fluctuation, TSH had only a delayed compensatory increase, and cholesterol increased during phenobarbital treatment. The delayed increase in TSH, despite persistent hypothyroxinemia, suggests that accelerated hepatic thyroxine elimination may not be the only effect of phenobarbital on the thyroid axis. There was no significant effect of phenobarbital on either of the adrenal function tests. With the methods employed, we did not find any effects of the drug on the hormonal equilibrium of the adrenal axis.

Determination of normal values using an automated coagulation timer for activated coagulation time and its application in dogs with hemophilia.

The present study was performed to determine normal values for the Medtronic HemoTec automated activated coagulation time (ACT) analyzer (Medtronic HemoTec Inc, Parker, CO, distributed in Switzerland by Convergenza AG, Vaduz, Liechtenstein), and to evaluate its ability to detect dogs with hemophilia. ACT was measured in 43 healthy dogs presented to the Companion Animal Hospital, University of Bern, Bern, Switzerland, with the Medtronic HemoTec ACT analyzer to determine normal values. The mean +/- 2 standard deviations (SDs) of the values obtained was defined as the normal range. ACT was measured 8-10 times on the same day in 6 dogs to determine repeatability. ACT also was measured in 11 dogs with hemophilia and compared with a conventional visual ACT measurement test and with the activated partial thromboplastin time (APTT). ACT values of the 43 dogs used to determine normal values ranged from 66.5 to 97.0 seconds (mean, 79.3 seconds; SD, 7.35 seconds; median, 78.5 seconds). A range of 64-95 seconds (mean +/- 2 SDs) was defined as the normal range for the tested device. Repeatability was poor (r = 0.256). ACT values measured with the automated device did not correlate with ACT values measured with a conventional visual test or with APTT Sensitivity of the test was 90.9%, specificity was 98.0%, and accuracy was 96.7%. Variability in the test results was large and may lead to incorrect results. The automated measurement device was not superior to the conventional visual method in evaluating dogs with hemophilia.

Cardiomyopathy in dystrophin-deficient hypertrophic feline muscular dystrophy.

A colony of cats affected with hypertrophic feline muscular dystrophy was used to study the occurrence of cardiomyopathy associated with dystrophin deficiency. Affected male and female cats, obligate carrier females, and unaffected healthy littermates were followed from 12 weeks of age into adulthood. Thoracic radiography, 2-D echocardiography, and 2-D-derived M-mode echocardiography were performed at 3-month intervals until 12 months of age and regularly thereafter. From 9 months of age, all affected cats had larger hearts than normal and carrier animals. Left ventricular wall thickness in systole and in diastole and interventricular septal thickness in systole were greater in affected cats 12 months and older when compared with normal or heterozygous animals (P < .05). The myocardium of affected cats was diffusely hypoechoic and thickened. Multiple hyperechoic foci were in the myocardium and papillary musculature. Shortening fraction was normal in all cats. Changes seen in carrier females included enlargement and hyperechogenicity of the papillary musculature after the age of 2 years. Gross and light microscopic examination revealed left ventricular wall thickening with multiple foci of mineralization in 2 of 5 hearts from dystrophin-deficient cats. Although approximately 10% of the normal dystrophin amount was present in the skeletal muscle, dystrophin could not be detected in the myocardium. Early onset concentric myocardial hypertrophy was present in all adult cats. Lesions were mainly localized in the myocardium of the left ventricular free wall and interventricular septum, papillary musculature, and the endocardium. Clinical signs of heart failure developed only infrequently in cats with hypertrophic feline muscular dystrophy.

Dystrophin deficiency causes lethal muscle hypertrophy in cats.

Two 5-month-old male Domestic Shorthair littermates showed general skeletal muscle hypertrophy, multifocal submucosal lingual calcification with lingual enlargement, and excessive salivation. Both cats had a reduced level of activity, walked with a stiff gait, and tended to "bunny hop" when they ran. These clinical features were similar to those of previously reported dystrophin-deficient cats. Using multiple dystrophin antibodies, we found that the cats described in this report also showed marked dystrophin deficiency. The histopathology was remarkable for hypertrophy and splitting of fibers, and progressive accumulation of calcium deposits within the muscle. There was little or no endomysial fibrosis at 2 years of age. The natural history of dystrophin-deficiency in cats has not been described: both previous cats had been euthanized at 2 years of age prior to experiencing any life-threatening problems. At 6 months of age, one of the new cats developed megaesophagus because of severe progressive hypertrophy of the diaphragmatic muscles. The diaphragm completely occluded the esophagus, and the cat was euthanized for humane reasons. The second cat remained in good condition until age 18 months when it developed acute renal failure attributed to severe prolonged dehydration and hyperosmolality. The cat recovered after receiving supportive treatment but was unable to maintain fluid homeostasis. The insufficient water intake was attributed to glossal hypertrophy and dysfunction. At age 2 years, the cat received regular subcutaneous injections of low-sodium fluids to maintain proper hydration. The clinical consequence of dystrophin deficiency in cats is lethal muscle hypertrophy. We have called the feline disease "hypertrophic feline muscular dystrophy" (HFMD).