Isoamylases in clinically normal dogs

Isoenzymes of canine serum amylase were evaluated by electrophoresis on cellulose acetate membranes in a discontinuous buffer system. Two isoamylase groups were identified in the serum of clinically normal dogs. Most of the serum amylase activity was the more cathodal isoamylase. The anodal isoamylase was rarely observed in serum from normal dogs and, when present, accounted for little of the enzymatic activity. Amylase activities of various tissues were determined and isoenzymes were identified. The anodal isoenzyme was found in the pancreas and uterus. The cathodal isoamylase had its origin mainly from the intestinal tract. Activities of other tissues were not greater than was serum amylase activity. Effects of feeding upon total serum amylase activity and isoenzyme composition also were examined over a period of 5 minutes to 7 hours. Feeding did not markedly alter the serum amylase activity.     

Synovial fluid cell counts and total protein concentration in clinically normal fetlock joints of young dromedarian camels

Twenty-seven 9-12 months old healthy male dromedarian camels were used to determine total nucleated leucocyte count (TNCC), absolute and percentages of polymorphonuclear (PMN) and mononuclear leucocytes, and total protein (TP) concentration in synovial fluid from grossly and radiographically normal fetlock joints. Arthrocentesis was performed bilaterally from the fetlock joints of the forelimbs and hindlimbs. Blood contaminated samples and samples obtained from grossly or radiographically abnormal joints were excluded. The mean +/- SD of TNCC in 108 samples of fetlock joint synovial fluids was 500 +/- 400 cells/microl. Monocytes/macrophages were the predominant cell type. There were no significant differences in mean TNCC, absolute numbers and percentages of various leucocytes and TP concentrations between the right and left fetlock joints of the forelimbs and hindlimbs or between the fetlock joints of the forelimbs and hindlimbs. The mean +/- SD of absolute numbers and percentages of various cell types were: PMN leucocytes 1 +/- 2 cells/microl (2%), lymphocytes 116 +/- 167 cells/microl (26%), and monocytes/macrophages 383 +/- 323 cells/microl (72%). The mean +/- SD of TP concentration was 2 +/- 1 g/dl.     

Ureterocolonic anastomosis in clinically normal dogs

Ureterocolonic anastomosis was evaluated in 13 clinically normal dogs. Urinary continence was maintained after surgery, and the procedure was completed without technique errors in all but 2 dogs. Three dogs died within 5 weeks (2 of undetermined causes and 1 of aspiration pneumonia and neurologic disease), and 1 dog was euthanatized 4 months after surgery because of neurologic signs. Two healthy dogs were euthanatized 3 months after surgery for light microscopic evaluation of their kidneys. Five dogs were euthanatized 6 months after surgery for light microscopic evaluation of their kidneys. Gastrointestinal and neurologic disturbances developed in 4 dogs at various postoperative intervals. Plasma ammonia concentration measured in 2 dogs with neurologic signs was increased. Plasma ammonia concentration measured in 5 dogs without neurologic signs was within normal limits. All 5 dogs, in which metabolic acidosis was diagnosed, had high normal or above normal serum chloride concentration. Serum urea nitrogen values were increased after surgery because of colonic absorption of urea. Serum creatinine concentration was increased in 1 dog 6 months after surgery. Individual kidney glomerular filtration rate was reduced in 38% (3/8) of the kidneys from 4 other dogs at 6 months after surgery. Of 5 dogs euthanatized at 3 to 4 months after surgery, 4 had bilateral pyelitis, and 1 had unilateral pyelonephritis. Six months after surgery, pyelonephritis was diagnosed in 40% (4/10) of the kidneys from 5 dogs. The ureterocolonic anastomosis procedure is a salvage procedure that should allow complete cystectomy. However, variable degrees of metabolic acidosis, hyperammonemia, and neurologic disease may result.     

Diseased dogs isoamylases in clinically normal and diseased dogs

Isoamylases in normal canine sera were separated on cellulose acetate membranes using a discontinuous buffer system without EDTA. Four peaks of amylase activity were present in 17 of 24 sera. Normal values were established. The majority of activity was present in Peak 4 (cathodal isoamylase). Tissue extracts of pancreas, duodenum, kidney, lung, testis, spleen and uterus-ovaries contained Peak 4 isoamylase. Liver and salivary gland lacked all isoamylase activity. Pancreas contained Peak 3 in addition to Peak 4 isoamylase. A tissue origin for Peaks 1 and 2 was not identified. An overall lack of resolution resulted from the inclusion of EDTA in the electrophoresis buffer system. This may account for previous findings suggesting that pancreatic amylase is not present in normal canine serum. An increase in the Peak 3 isoamylase was present in dogs with pancreatitis while dogs with pancreatic atrophy had a decrease in all isoamylases. Total amylase activity was significantly (p < 0.05) decreased in dogs with pancreatic atrophy.     

Impedance audiometric measurements in clinically normal dogs

OBJECTIVE: To measure impedance audiometric values in clinically normal dogs that were sedated or anesthetized, evaluate effects of ear flushing on tympanometric measurements, and determine effects of performing acoustic reflex testing in a sound-attenuated room. ANIMALS: 35 mixed-breed and purebred client-owned dogs and 21 laboratory-bred Beagles. PROCEDURES: Tympanometry and acoustic reflex testing were performed on 27 mixed-breed and purebred dogs under isoflurane anesthesia in a non-sound-attenuated room and 21 Beagles under sedation in a sound-attenuated room. Tympanometry was performed on 8 mixed-breed dogs under halothane anesthesia before and after ear canal flushing. RESULTS: Among impedance audiometric values, ear canal volume and compliance peak were smaller in Beagles than in mixed-breed dogs; differences among other values were not detected. Ear canal volume was dependent on body weight. Differences were not found for tympanometric values measured before and after ear canal flushing. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study established reference range values for impedance audiometric measurements in clinically normal dogs under isoflurane anesthesia or sedation. Acoustic reflex testing does not need to be performed in a sound-attenuated room. The ear canals of clinically normal dogs can be flushed prior to performing tympanometry without altering the results. Impedance audiometry may be a useful noninvasive procedure for the diagnosis of otitis media in dogs.     

Telomerase activity in clinically normal dogs and dogs with malignant lymphoma

OBJECTIVES: To determine whether telomerase activity was present in lymph nodes, buffy coat, and serum samples from dogs with malignant lymphoma (ML) and in liver, lymph node, buffy coat, and serum samples from clinically normal dogs SAMPLE POPULATION: Tissue specimens and blood samples were obtained from 11 clinically normal adult dogs (age range, 1 to 4 years) and 14 client-owned dogs with ML. PROCEDURE: The telomere repeat amplification protocol assay was used to quantify telomerase activity in the tissues from clinically normal dogs and dogs with ML. RESULTS: Of 11 clinically normal dogs, 8 had lymph node samples, 5 had liver samples, and 1 had buffy coat samples with detectable telomerase activity. None of the serum samples from the clinically normal dogs had detectable telomerase activity. Of 14 dogs with ML, 9 had lymph node samples, 3 had buffy coat samples, and 1 had serum samples with measurable telomerase activity. CONCLUSIONS AND CLINICAL RELEVANCE: Telomerase activity was not specific to tumor cells and overlapped with that found in cells from clinically normal dogs. Telomerase activity in neoplastic lymph nodes was not substantially different from that found in lymph nodes from clinically normal dogs. The determination of telomerase activity cannot be used as a sole diagnostic test for cancer. Therapeutic modalities directed toward the telomerase enzyme may not be feasible in dogs, because somatic tissues from clinically normal dogs possess variable amounts of telomerase activity.     

Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis

Background: Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. Objectives: The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Methods: Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–Whitney U test. Results: Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α₁‐globulin (1.9–3.1 g/L), α₂‐globulin (5.3–8.7 g/L), β₁‐globulin (2.8–7.3g/L), β₂‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65). Conclusion: Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.     

Urine metabolite values in fed and nonfed clinically normal beagles.

Twenty-four-hour excretion of urine metabolites was determined in 33 clinically normal Beagles during periods of consumption of a standard diet and when food was withheld. The goal was to determine normal canine values for urine analytes incriminated in the genesis of calcium oxalate uroliths. During periods when dogs consumed food, dairy urinary excretion of calcium, uric acid, sodium, potassium, magnesium, ammonium, and hydrogen ions were significantly (P = 0.0004, 0.0038, 0.001, 0.0001, 0.0004, 0.0001, and 0.024, respectively) higher than when food was withheld. Urinary excretion of phosphorus, oxalate, and citrate were not significantly different between samples obtained during periods of food consumption and when food was withheld. Male dogs excreted significantly higher quantities of urine oxalate than females during fed (P = 0.003) and nonfed (P = 0.003) conditions. When food was withheld, urinary uric acid excretion was significantly higher in males than females (P = 0.01). Females excreted significantly more urine calcium than males when food was withheld (P = 0.003). Our results indicated that dietary conditions influence the quantity of sodium, potassium, calcium, magnesium, and uric acid excreted in the urine of clinically normal dogs; therefore, dietary conditions should be considered when measuring the concentration of these analytes in urine.     

Variability of estradiol concentration in normal dogs

Estradiol concentrations are evaluated in canine serum as part of an adrenal panel used to diagnose atypical Cushing’s syndrome and other endocrine abnormalities. Estradiol concentrations are often elevated in dogs without clinical signs of hyperestrogenism, and the significance of this elevation is unknown. The purpose of this study was to estimate the variation in estradiol concentrations in normal dogs. Ten neutered male and female dogs were enrolled in the study. Blood was collected from each dog at 2 h intervals, four times during a given day. This was repeated approximately 1 (week 2) and 5 weeks later (week 6). There was no attempt for a given dog to be started at the exact time or day each week. Results showed that estradiol concentrations ranged from 44.6 to 120.3 pg/mL with a mean of 70.4 pg/mL, which is greater than the upper limit of normal for our laboratory (69 pg/mL). The mean difference between the highest and lowest concentrations for each dog was 28.8 pg/mL, with a range of 12.5–53.5 pg/mL. Mean estradiol concentrations from week 6 (63.2 pg/mL) were significantly lower than those from week 1 (71.4 pg/mL; P = 0.015) and week 2 (76.5 pg/mL; P = 0.0004). These data show a wide range of variability in estradiol concentration both within and between dogs and that these measurements often exceed the normal ranges established by the laboratory. Therefore, diagnosis of hyperestrogenism or atypical Cushing’s syndrome based on increased estradiol concentrations should require compatible clinical presentation of hyperestrogenism together with elevated serum estradiol.     

Osmotic fragility of erythrocytes in clinically normal dogs and dogs infected with parasites

The osmotic fragility of the erythrocytes was measured in blood samples collected from randomly selected healthy and infected dogs at a dogs' rescue shelter. The dogs were classified into six groups on the basis of the final diagnoses from clinical, post mortem and laboratory findings. The minimum (less than 5 per cent) and maximum (more than 90 per cent) haemolysis of the erythrocytes of the clinically normal dogs (group 1), occurred in 0·60 per cent and 0·30 per cent solutions of sodium chloride (NaCI). For the non-anaemic hookworm-infected dogs (group 2a) the respective values were 0·8 per cent and 0·4 per cent NaCl, and for the anaemic hookworm-infected dogs (group 2b) they were 0·85 per cent and 0·5 per cent NaCl, respectively. The erythrocytes from dogs with Babesia canis (group 3), concurrent hookworm and B canis (group 4) and Ehrlichia canis infections (group 5) had minimum haemolysis in 0·75 per cent NaCl and maximum haemolysis at between 0·20 per cent and 0·35 per cent NaCI solutions. The derivative fragiligrams for groups 2a, 2b, 3 and 4 were shifted to the left, whereas the fragiligram for group 5 was similar to that for the clinically normal dogs (group 1). The left shift for the hookworm-infected dogs was due to the increased osmotic fragility of a minor sub-population of the erythrocytes, but for the dogs, infected with B canis major sub-populations of the erythrocytes had an increased osmotic fragility.     

Nuclear scintigraphic assessment of liver size in clinically normal dogs

Measurement of liver size was made on nuclear scintigraphic images obtained from 16 clinically normal, anesthetized dogs in ventral, dorsal, right and left lateral, and left dorsal oblique positions after administration of technetium 99m-sulfur colloid. Linear measurements of liver length and width were made from each scintiscan, and liver surface area was determined by setting a region of interest manually and by use of a computer count of pixels above a minimal intensity (threshold method). All linear measurements had a statistically significant (P less than 0.05) correlation with liver and body weight, with the exception of the measurements of liver width made on dogs in dorsal and left lateral dorsal oblique positions. The highest correlation (r = 0.89) between the scintigraphic measurements and liver weight was the multiplication of measurements of liver width and length from the right lateral view. Although all area determinations were significantly (P less than 0.05) correlated with liver and body weight, for most views, the manual method of determining the region of interest had a slightly better correlation with the liver weight than did the threshold method.     

Nitrogen balance in clinically normal dogs receiving parenteral nutrition solutions

OBJECTIVE: To determine nitrogen balance in clinically normal dogs receiving parenteral nutrition solutions. ANIMALS: 8 clinically normal female Beagles. PROCEDURE: Dogs were randomly assigned to receive 4 treatments in random order. Treatment A consisted of IV administration of nonlactated Ringer's solution. Treatments B, C, and D consisted of IV administration of isocaloric parenteral solutions containing 0, 1.36, and 2.04 g of amino acids/kg of body weight/d, respectively, for 7 consecutive days. Urine and feces were collected on days 5, 6, and 7 of each treatment period, and Kjeldahl analysis was used to determine nitrogen balance. RESULTS: Mean nitrogen balance was negative with treatments A and B but was not significantly different from 0 with treatments C and D. Dogs had the lowest nitrogen balance values and lost the most weight while receiving treatment A. Dogs were able to conserve protein and had higher nitrogen balance values when receiving treatment B, compared with treatment A. Dogs lost the least amount of weight while receiving treatment D. Regression analysis indicated that an IV amino acid intake of 2.32 g/kg/d (95% confidence interval, 2.00 to 2.81 g/kg/d), as supplied by the commercial product used in this study, would result in zero nitrogen balance in clinically normal dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that IV amino acid requirement of clinically normal dogs is approximately 2.3 g/kg/d.     

Effect of mitotane on pituitary corticotrophs in clinically normal dogs

OBJECTIVE: To evaluate the effects of mitotane administration on the function and morphology of pituitary corticotrophs in clinically normal dogs. ANIMALS: 12 clinically normal adult Beagles. PROCEDURES: Dogs were randomly assigned to the control group or the mitotane treatment group. In mitotane treatment group dogs, mitotane was administered for 1 month. In both groups, ACTH stimulation testing and corticotrophin-releasing hormone (CRH) stimulation testing were performed. Magnetic resonance imaging (MRI) of the pituitary gland and brain was performed in mitotane treatment group dogs before and after administration of mitotane. After CRH stimulation testing and MRI, dogs were euthanatized and the pituitary gland and adrenal glands were excised for gross and histologic examination. RESULTS: ACTH concentrations in mitotane treatment group dogs were significantly higher than in the control group dogs following CRH stimulation. Magnetic resonance imaging revealed that pituitary glands were significantly larger in treatment group dogs after administration of mitotane, compared with before administration. On gross and histologic examinations, the adrenal cortex was markedly atrophied. Immunohistochemistry revealed hypertrophy of corticotrophs in pituitary glands of mitotane treatment group dogs. CONCLUSIONS AND CLINICAL RELEVANCE: These findings indicate that inhibition of the adrenal cortex by continuous administration of mitotane leads to functional amplification and morphologic enhancement of corticotrophs in clinically normal dogs. In instances of corticotroph adenoma, hypertrophy of individual corticotrophs induced by mitotane may greatly facilitate enlargement of the pituitary gland and increases in ACTH secretion.     

Quantitative noninvasive assessment of liver size in clinically normal dogs

Measurements of liver size were made from radiographs of 16 clinically normal anesthetized dogs. Two measurements were made from each of 3 views: right and left lateral and ventrodorsal. Each measurement was correlated with liver weight and volume. Liver weight and volume were also correlated with both measurements from the same radiograph multiplied together. All measurements, with the exception of ventrodorsal depth, correlated significantly (P less than 0.05) with liver weight and volume. The multiplication of the 2 measurements made from the right lateral radiograph correlated most highly with liver weight. These data were then compared with those from similar studies involving liver measurements obtained from nuclear scintigraphic and ultrasonographic scans on the same dogs. The radiographic measurements made from the right lateral view had the highest correlation with liver weight, followed by the nuclear scintigraphic measurements made from the same position.     

Electromyographic and esophagomanometric findings in clinically normal dogs and in dogs with idiopathic megaesophagus.

Electromyography of 12 clinically normal dogs and 7 dogs with idiopathic megaesophagus revealed trains of positive sharp waves in the muscles of facial expression and in the lingual muscles of both groups. Positive waves are usually indicative of motor-unit disease; however, they are clinically insignificant in these muscles. Positive sharp waves were detected in the esophageal muscle of one dog with congenital megaesophagus. Esophageal electromyograms obtained in a dog with congenital megaesophagus and in 2 clinically normal dogs were normal. Resting caudal esophageal sphincter pressure was similar in both clinically normal dogs (mean, 22.3 mm of Hg; range, 15--37 mm of Hg) and in dogs with congenital or acquired idiopathic megaesophagus (mean, 29.6 mm of Hg; range, 20--50 mm of Hg).     

Fluorophotometric determination of aqueous humor flow rate in clinically normal dogs

OBJECTIVE: To determine aqueous humor flow rate in clinically normal dogs, using fluorophotometry. ANIMALS: 20 clinically normal Beagles. PROCEDURE: A study was performed on 5 dogs to establish an optimal protocol for fluorophotometric determination of aqueous humor flow rate. This protocol then was used to measure aqueous humor flow rate in 15 dogs. Corneas were loaded with fluorescein by topical application, and corneal and aqueous humor fluorescein concentrations were measured 5, 6.5, and 8 hours after application. Concentration-versus-time plots were generated, and slopes and ratios of the fluorescein concentration in the cornea and aqueous humor from these graphs were used to calculate flow rates. Calculations were performed by use of automated software provided with the fluorophotometer and by manual computation, and the 2 calculation methods were compared. RESULTS: The protocol established for the 5 dogs resulted in semilogarithmic and parallel decay of corneal and aqueous humor concentrations. Manually calculated mean +/- SD aqueous humor flow rates for left, right, and both eyes were 5.58 +/- 2.42, 4.86 +/- 2.49, and 5.22 +/- 1.87 microl/min, respectively, whereas corresponding flow rates calculated by use of the automated software were 4.54 +/- 3.08, 4.54 +/- 3.10, and 4.54 +/- 2.57 microl/min, respectively. Values for the left eye were significantly different between the 2 computation methods. CONCLUSIONS AND CLINICAL RELEVANCE: Aqueous humor flow rates can be determined in dogs, using fluorophotometry. This technique can be used to assess pathologic states and medical and surgical treatments that alter aqueous humor dynamics.     

Ultrasonographic and laboratory screening in clinically normal mature golden retriever dogs

Wellness and pre-anesthetic screening of blood and urine of geriatric companion animals are routinely recommended. In addition, there are occasional references to the use of imaging in clinically normal geriatric patients. However, the utility of wellness testing is not known, and there is limited information regarding the value of pre-anesthetic testing. Wellness testing, including complete blood cell count, biochemical profile, urinalysis, and abdominal ultrasound, was performed on 53 clinically normal, mature golden retriever dogs. Laboratory analysis revealed abnormalities in 54.7% (29/53) of the dogs. Abdominal ultrasound screening demonstrated abnormalities in 64.2% (34/53) of the dogs. As only a small number of dogs had follow-up diagnostic testing available, the significance of these abnormalities is unknown. Further study involving a larger cohort of animals and analysis of follow-up data is necessary to determine the utility of laboratory and imaging studies in clinically normal geriatric patients.