In vitro effects of bethanechol on specimens of intestinal smooth muscle obtained from the duodenum and jejunum of healthy dairy cows

OBJECTIVE: To describe the in vitro effects of bethanechol on contractility of smooth muscle preparations from the small intestines of healthy cows and define the muscarinic receptor subtypes involved in mediating contraction. SAMPLE POPULATION: Tissue samples from the duodenum and jejunum collected immediately after slaughter of 40 healthy cows. PROCEDURES: Cumulative concentration-response curves were determined for the muscarinic receptor agonist bethanechol with or without prior incubation with subtype-specific receptor antagonists in an organ bath. Effects of bethanechol and antagonists and the influence of intestinal location on basal tone, maximal amplitude (A(max)), and area under the curve (AUC) were evaluated. RESULTS: Bethanechol induced a significant, concentration-dependent increase in all preparations and variables. The effect of bethanechol was more pronounced in jejunal than in duodenal samples and in circular than in longitudinal preparations. Significant inhibition of the effects of bethanechol was observed after prior incubation with muscarinic receptor subtype M(3) antagonists (more commonly for basal tone than for A(max) and AUC). The M(2) receptor antagonists partly inhibited the response to bethanechol, especially for basal tone. The M(3) receptor antagonists were generally more potent than the M(2) receptor antagonists. In a protection experiment, an M(3) receptor antagonist was less potent than when used in combination with an M(2) receptor antagonist. Receptor antagonists for M(1) and M(4) did not affect contractility variables. CONCLUSIONS AND CLINICAL RELEVANCE: Bethanechol acting on muscarinic receptor sub-types M(2) and M(3) may be of clinical use as a prokinetic drug for motility disorders of the duodenum and jejunum in dairy cows.     

Does the computed tomographic appearance of the lung differ between young and old dogs?

In computed tomographic (CT) images of humans, decreased lung attenuation, bronchial dilation, and/or thickening, air trapping, cysts, and thickened interlobular septa have been associated with increasing age. To determine if there are differences in the CT appearance of the lungs of young and old dogs that could affect interpretation of diagnostic studies, pulmonary CT images of dogs with conditions unrelated to the thorax were reviewed retrospectively in a case–control study. Computed tomography studies of 42 young dogs (range 0.3–4.8 years) and 47 old dogs (range 9–15.1 years) were jumbled and reviewed by an observer blinded to dog age. Computed tomography was performed under sedation in 62 (70%) dogs and under general anesthesia in 27 (30%). Heterotopic bone was more prevalent (62% vs. 14%) in old dogs. Lung collapse was significantly associated with old age, greater body weight, and anesthesia. There were no significant differences in median lung attenuation or occurrence of ground glass pattern, cysts, bronchial thickening, bronchial dilation, or degree of tracheal calcification. No examples of reticular pattern, emphysema, pleural thickening, or septal thickening were observed in any dog. Despite previous studies describing age‐related changes in the radiographic appearance of the lungs of old dogs, it appears that there are minimal observable differences in CT images. Old dogs are more likely to have visible foci of heterotopic bone and may be more prone to lung lobe collapse than young dogs, but neither of these differences should contribute to misdiagnosis of pulmonary disease.     

ULTRASONOGRAPHIC FINDINGS IN CHOLECYSTOKININ-INDUCED PANCREATITIS IN DOGS

Pancreatitis can be induced in dogs by intravenous infusion of supraphysiologic doses of synthetic cholecystokinin octapeptide (CCK-8). Eight anesthetized female beagles were given either CCK-8 or saline in a randomized, blinded fashion and had abdominal ultrasonography to study the development of pancreatic lesions. Pancreatic lesions were apparent 2 hours after the start of CCK-8 infusion and included swelling, interlobular and subcapsular fluid accumulation and patchy hypoechogenicity of the gland parenchyma. CCK-8 had a variable effect on gallbladder emptying. No signs of biliary obstruction or duodenal lesions were identified. Dogs given CCK-8 had severe edematous pancreatitis at necropsy six hours after the start of the infusion. Compared to the oleic acid model of pancreatitis, CCK-8 infusion induces a rapid onset of pancreatitis and an ultrasonographic appearance that reflects diffuse edema of the gland. The short time-scale of the experiment may account for the relative absence of secondary lesions affecting the biliary tract or duodenum compared to the naturally-occurring disease.     

Diagnostic and public health investigation of Mycobacterium tuberculosis infection in a dog in Ontario,Canada

A 4-y-old, female mixed-breed dog was presented to the Ontario Veterinary College for further evaluation of multiple pulmonary and hepatic masses, intrathoracic lymphadenitis, and recent development of a pyogranulomatous pleural effusion. Along with other comprehensive tests, a thoracic lymph node biopsy was performed, and Mycobacterium tuberculosis complex infection was confirmed by real-time PCR. The dog’s condition declined post-operatively, and euthanasia was elected. Postmortem examination confirmed severe granulomatous pneumonia, hepatitis, intrathoracic and intraabdominal lymphadenitis, omentitis, and nephritis. Line-probe assays performed on samples collected postmortem confirmed the species as M. tuberculosis. 24-loci MIRU-VNTR genotyping, spoligotyping, and whole-genome sequencing revealed relations to known human isolates, but no epidemiologic link to these cases was investigated. Given the concern for potential human exposure during this animal’s disease course, a public health investigation was initiated; 45 individuals were tested for M. tuberculosis exposure, and no subsequent human infections related to this animal were identified. Our case highlights the need for more readily available, minimally invasive testing for the diagnosis of canine mycobacteriosis, and highlights the ability of canid species to act as potential contributors to the epidemiology of M. tuberculosis infections.     

MAGNETIC RESONANCE IMAGING OF CEREBRAL CORTICAL NECROSIS (POLIOENCEPHALOMALACIA) IN A DOG

A 3-year-old neutered female mixed breed dog was examined because of severe, generalized seizure activity, tetraparesis, and encephalopathic signs. Cerebrospinal fluid (CSF) evaluation was unremarkable except for a mild increase in protein. Serum and CSF titers for infectious diseases were negative. Magnetic resonance (MR) imaging examination of the brain was performed and lesions were found within the cerebral gray matter of the temporal and parietal lobes. The lesions had increased signal intensity on T1, T2, and proton density-weighted images. There was mild inhomogeneous enhancement following intravenous contrast medium administration. Neurologic status improved and the seizures were well controlled, but the dog never regained normal mentation and euthanasia was performed 10 weeks after initial evaluation. At necropsy, severe cerebral cortical necrosis was found in the regions corresponding to the lesions seen on MR imaging examination. Large numbers of fat-containing macrophages (gitter cells) were found within these areas, and are thought to be responsible for the characteristic hyperintensity seen on the MR images.     

Evaluation of cystatin C as an endogenous marker of glomerular filtration rate in dogs

Cystatin C is a cysteine protease inhibitor produced by all nucleated cells. It is freely filtered by the glomerulus and is unaffected by nonrenal factors such as inflammation and gender. Because of greater sensitivity and specificity, cystatin C has been proposed to replace creatinine as a marker of glomerular filtration rate (GFR) in humans. The aims of this study were to validate an automated assay in canine plasma and to evaluate the usefulness of cystatin C as a marker of GFR in dogs. Western blotting was used to demonstrate cross-reactivity of an anti-human cystatin C antibody. An immunoturbidimetric assay was used to detect cystatin C in 25 clinically healthy dogs and 25 dogs with renal failure. Mean cystatin C concentration in the healthy dogs and the dogs with renal failure was 1.08 +/- 0.16 mg/L and 4.37 +/- 1.79 mg/L respectively. Intra- and interassay variability was <5%. The assay was linear (r = .974) between 0.14 and 7.53 mg/L. Both cystatin C and creatinine concentrations were measured in banked, frozen serum from 20 remnant kidney model dogs and 10 volume-depleted dogs for which GFR measurements by exogenous creatinine clearance had been determined previously. In the remnant kidney model, cystatin C was better correlated with GFR than creatinine (r = .79 versus .54) but was less well correlated with GFR in volume-depleted dogs (r = .54 versus .95). GFR measurements were repeated in the remnant kidney model dogs 60 days after initial GFR measurements. At this time, cystatin C and creatinine concentrations correlated equally well with GFR (r = .891 versus .894, respectively). Cystatin C concentration is a reasonable alternative to creatinine for screening dogs with decreased GFR due to chronic renal failure.     

DOES MEASUREMENT OF SMALL INTESTINAL DIAMETER INCREASE DIAGNOSTIC ACCURACY OF RADIOGRAPHY IN DOGS WITH SUSPECTED INTESTINAL OBSTRUCTION?

The ratio between maximal small intestinal (SI) diameter and the height of the body of the fifth lumbar vertebra (L5) in radiographs has been reported as a diagnostic test in dogs with suspected intestinal obstruction. In order to assess the effect of the SI/L5 ratio on the accuracy of radiographic diagnosis of intestinal obstruction, lateral abdominal radiographs of 37 dogs with small intestinal obstruction and 48 nonobstructed dogs were mixed and examined independently by six observers who were unaware of the final diagnosis and who represented a range of experience. Observers first examined radiographs subjectively and stated the likelihood of obstruction (definitely not, probably not, equivocal, probably, definitely). Observers subsequently reexamined the radiographs, determined the SI/L5 ratio, and again stated the likelihood of obstruction. The most frequent cause of obstruction was foreign body (29/37, 78%). Dogs with SI obstruction had a significantly larger median SI/L5 ratio than nonobstructed dogs (P = 0.0002). Using an SI/L5 ratio of 1.7 for diagnosis of intestinal obstruction, sensitivity and specificity were 66%. Use of the SI/L5 ratio was not associated with increased accuracy of diagnosis for any observer, regardless of experience, hence this test may have no diagnostic impact.     

Histological and morphometric lesions in the pre-clinical,developmental phase of insulin-induced laminitis in Standardbred horses

Lamellar pathology in experimentally-induced equine laminitis associated with euglycaemic hyperinsulinaemia is substantial by the acute, clinical phase (～48 h post-induction). However, lamellar pathology of the developmental, pre-clinical phase requires evaluation. The aim of this study was to analyse lamellar lesions both qualitatively and quantitatively, 6, 12 and 24 h after the commencement of hyperinsulinaemia. Histological and histomorphometrical analyses of lamellar pathology at each time-point included assessment of lamellar length and width, epidermal cell proliferation and death, basement membrane (BM) pathology and leucocyte infiltration. Archived lamellar tissue from control horses and those with acute, insulin-induced laminitis (48 h) was also assessed for cellular proliferative activity by counting the number of cells showing positive nuclear immuno labelling for TPX2.Decreased secondary epidermal lamellar (SEL) width and increased histomorphological evidence of SEL epidermal basal (and supra-basal) cell death occurred early in disease progression (6 h). Increased cellular proliferation in SELs, infiltration of the dermis with small numbers of leucocytes and BM damage occurred later (24 and 48 h). Some lesions, such as narrowing of the SELs, were progressive over this time period (6–48 h). Cellular pathology preceded leucocyte infiltration and BM pathology, indicating that the latter changes may be secondary or downstream events in hyperinsulinaemic laminitis.     

Tono-Pen XL calibration curves for cats, cows and sheep

The objective of this study was to provide calibration curves for correcting intraocular pressure (IOP) measurements obtained using the Tono-Pen XL tonometer in cats, cows and sheep. Twelve eyes from 9 cats, 13 eyes from 7 cows, 10 eyes from 5 sheep were used. The anterior chamber of the eye was cannulated in vivo, in situ (immediately post mortem) or ex vivo with a fine needle and IOP was varied from 10 to 90 mmHg in steps of 10 mmHg by adjusting the height of a saline reservoir connected to the needle. For each pressure setting, several readings of IOP were made using the tonometer. The relationship between Tono-Pen reading and manometer setting was linear over the full range of measurement. However, the slope of the data regression line deviated significantly from 1 and indicated that the instrument systematically underestimated IOP. For cats the average slope was 0.62 and for cows and sheep it was 0.72 and 0.69, respectively. For the latter animals, the regression line also had a nonzero intercept of approximately 4.5 mmHg. Similar results were obtained from in vivo and ex vivo eyes and with different Tono-Pen XL tonometers. Although developed for use on humans, the Tono-Pen XL can provide reproducible and accurate measurement of IOP in cats, cows and sheep when suitably calibrated by manometry. The calibration curves provided here, and by implication those reported for other animals using this tonometer, differ in slope from those measured with earlier models of the Tono-Pen. The reproducibility of the curves we obtained implies that they can be used to correct IOP readings from the Tono-Pen XL when manometry is not possible.