Effect of dietary protein on lean body wasting in dogs: correlation between loss of lean mass and markers of proteasome-dependent proteolysis.

To determine the effect of dietary protein intake on lean body wasting in adult canines a study was undertaken to investigate the Ubiquitin Proteasome (UP) pathway and concurrent changes in lean and fat body mass of canines fed variable sources and concentrations of dietary protein. Purpose-bred, intact female canines (56) between the ages of 2 and 3 years were fed either 12 or 28% protein diet for 10 weeks. Each diet contained variable amounts of corn gluten meal and chicken protein sources in ratios of 100 : 0, 67 : 33, 33 : 67 and 0 : 100 per cent (w/w), respectively. All diets were isocaloric with calories coming from protein : fat : carbohydrate at the respective ratios of 12 : 40 : 48% for the 12% diets, and 28 : 40 : 32% for the 28% diets. Standard dual energy X-ray absorptiometry was performed to assess total body lean and fat mass at weeks 0 and 10 of the dietary trial. Muscle biopsies were also taken and processed for protein determination and standard gel electrophoresis with subsequent Western blotting for 20S proteasome and PA700 regulatory cap subunit p31. Statistical analysis revealed a moderate degree of correlation between increasing quantities of corn gluten, which is low in essential amino acids (i.e. lysine, tryptophan), and increasing loss of lean body mass over the 10-week study (R = 0.56). Furthermore, a moderate degree of correlation was observed between increasing concentrations of corn gluten protein and decreased expression of the p31 subunit of the 26S proteasome (R = 0.49). Additionally, the dogs consuming the 12% protein diets had a significant increase in fat mass regardless of the protein source. These findings suggest that lean body wasting in adult canines can be associated with the consumption of low protein diets consisting of predominantly corn gluten, which is likely due to imbalances or subclinical deficiencies of specific essential amino acids, and that low protein diets may augment accumulation of adipose tissue. Although the mechanisms remain unclear, alteration of molecular targets of skeletal muscle proteolysis, specifically involving the UP pathway occur.

Effect of postexercise carbohydrate supplementation on muscle glycogen repletion in trained sled dogs.

OBJECTIVE: To evaluate the effect of immediate postexercise carbohydrate supplementation on muscle glycogen (MG) repletion during the first 4 hours of recovery in sled dogs. ANIMALS: 24 Alaskan Huskies. PROCEDURE: Dogs were assigned to 1 of 3 treatment groups, and a muscle biopsy specimen was obtained 1 hour before and immediately (group A) or 4 hours (groups B and C) after a 30-km run. Immediately after exercise, dogs in group A and group C were given water; dogs in group B were given a glucose polymer solution (1.5 g/kg of body weight) in water. RESULTS: At 4 hours after exercise, MG concentration was significantly greater in group-B than in group-C dogs; the value in group-C dogs was not different from the value in group-A dogs immediately after exercise. Assuming similar rates of glycogen depletion between treatment groups, during the first 4 hours of recovery, group-B dogs replaced 49% of the glycogen used during exercise. Plasma glucose concentration was significantly greater in group-B than in group-A and group-C dogs at 100 minutes after exercise. CONCLUSIONS: Immediate postexercise carbohydrate supplementation in sled dogs leads to increased glucose concentration, which in turn promotes more rapid rate of MG repletion in the first 4 hours of recovery than is observed in dogs not given supplements. CLINICAL RELEVANCE: For dogs running in multiple heats on a single day or over several consecutive days, immediate postexercise carbohydrate supplementation may promote more rapid and complete recovery between bouts of exercise.

Effect of diet and training on muscle glycogen storage and utilization in sled dogs.

Two groups of eight Alaskan huskies fed either a high-fat (HFD; 60% kcal from fat and 15% kcal from carbohydrate) or a high-carbohydrate diet (HCD; 60% kcal from carbohydrate and 15% kcal from fat) performed standard aerobic (1 h at 4 m/s on a 0% slope) and anaerobic (3 min at 6.7 m/s on a 10% slope) tests before and after training. Before and immediately after each exercise test, venous blood samples were collected and analyzed for lactate and pyruvate, and muscle biopsies were obtained under local anesthesia from the semitendinosus muscle and analyzed for total muscle glycogen (TMG) concentration. Training was associated with a significant increase in preexercise TMG in both diet groups; this effect was most marked in the HCD. There was no effect of diet or training on TMG utilization during the aerobic tests. The rate of TMG utilization during the anaerobic tests was between 20 and 40 times greater than that measured during the aerobic tests. The pre- to postexercise change in TMG was dependent on preexercise TMG in the HCD and HFD for both anaerobic tests (HCD: P < 0.01, r = 0.81; HFD: P < or = 0.03, r = 0.66). It is concluded that the increased glycogen storage associated with the HCD was more than offset by the more rapid rate of glycogen utilization in this group. HFD facilitated carbohydrate sparing during intense exercise and should thus be a better dietary strategy for endurance in sled dogs.

New approach to percutaneous muscle biopsy in dogs.

The size and quality of muscle specimens obtained by use of a percutaneous biopsy technique were studied. All biopsies were performed under local anesthesia, using an 11-gauge biopsy needle. The mean +/- SEM size of specimens obtained from 128 biopsies of the semitendinosus muscles of 16 Alaskan Huskies was 23.8 +/- 4.4 mg. All biopsy specimens were of sufficient quality to permit histochemical differentiation of the fiber types by use of myosin ATPase staining. An additional 8 biopsy specimens were obtained from 1 dog and analyzed for muscle glycogen content. These specimens contained 50.6 +/- 7.2 mmol of glucose/kg of muscle wet weight. This modified biopsy procedure was free of notable complications, and repeatable use produced specimens of adequate size and quality for histologic and biochemical analysis. It is concluded that this procedure is a safe and reliable alternative to open biopsy for diagnosis and management of neuromuscular, metabolic, and nutritional myopathies.

Use of scintimetry to assess effects of exercise and polysulfated glycosaminoglycan on equine carpal joints with osteochondral defects.

Combined blood pool and delayed images produced by use of 99mTc-methylene diphosphonate (99mTcMDP) were evaluated as an objective measurement of the response of equine joints with osteochondral defects to postoperative exercise and intra-articularly administered polysulfated glycosaminoglycan (PSGAG). Osteochondral defects (approx 2.4 x 0.9 cm) were induced arthroscopically in the dorsodistal radial carpal bones of 18 ponies. These ponies were randomized (while balancing for age [range 2 to 15; median, 5.0; mean, 5.1 years]) to 2 treatment groups. Nine ponies were assigned to be exercised, and 9 were stall-rested. Six ponies in each group were administered PSGAG (250 mg) in 1 joint (medicated) and lactated Ringer's solution (LRS) in the contralateral joint. The 3 remaining ponies in each group were administered LRS in both joints (nonmedicated). Medication was given at surgery, then weekly for 4 weeks. The exercise protocol (begun at postoperative day 6 and conducted twice daily) started with 30 minutes walking (approx 0.7 m/s), and, by postoperative month 3, the ponies were being walked for 15 minutes and trotted (approx 1.6 m/s) for 25 minutes. Simultaneous dorsal images of both carpi were made 2 to 3 minutes after IV administration of 99mTcMDP (blood pool image) and 90 to 120 minutes later (delayed image). Scintimetry, in counts per minute per pixel per millicurie, was done before, and at 1, 2, 4, 8, 10, 13, and 17 weeks after surgery, prior to euthanasia. Radionuclide uptake on blood pool images decreased faster than that on delayed images, in which uptake remained high for 17 weeks. This indicated that bone was metabolically active for at least 17 weeks after surgery. Exercise significantly (P < 0.05) decreased uptake on the blood pool images of medicated joints up to 1 month after surgery. Thus, exercise (in the presence of PSGAG) probably had a transient, beneficial effect on soft tissues of the joint. Exercise, without PSGAG, promoted increased bone remodeling, because the highest uptake on delayed images was observed in exercised, nonmedicated ponies up to 3 months after surgery. This was consistent with development of osteoarthritis in these ponies. Medication alone stimulated bone remodeling, and data indicated that an identical effect may take place in contralateral LRS-injected joints, because of systemic circulation of the drug. However, the combination of exercise and medication appeared to moderate the independent effects of each. The combination of exercise and medication in individual joints resulted in notably (P < 0.05) decreased bone remodeling.(ABSTRACT TRUNCATED AT 400 WORDS)

Functional analysis of ectopic parathyroid activity in cats.

The ability of ectopic parathyroid tissue to support calcium homeostasis was evaluated by measuring serum concentrations of calcium, phosphorus, albumin, magnesium, and parathyroid hormone before and for 12 weeks after bilateral thyroparathyroidectomy in 14 cats. During the immediate postoperative period, significant decrease was observed in serum calcium, magnesium, and parathyroid hormone (PTH) concentrations. Serum PTH concentration remained subnormal and did not significantly increase during the 12-week observation period. Despite persistent hypoparathyroidism, serum calcium and magnesium concentrations gradually increased. Ectopic parathyroid tissue is not capable of maintaining normal serum calcium concentration immediately after thyroparathyroidectomy. Serum calcium concentration gradually normalizes after thyroparathyroidectomy, apparently by means of a PTH-independent mechanism.

Dietary taurine deficiency and dilated cardiomyopathy in the fox.

Taurine deficiency has been implicated as a potential cause of dilated cardiomyopathy. However, the relationship between taurine and myocardial function is presently unclear. The purpose of this study was to determine whether dilated cardiomyopathy in the fox is associated with dietary taurine deficiency. A total of 68 foxes from farms with a history of death caused by dilated cardiomyopathy and 14 foxes from a farm with no history of dilated cardiomyopathy were studied. Dilated cardiomyopathy was diagnosed by echocardiography in 48% of the foxes from one farm with a positive history and in none of the foxes from the control farm. Foxes less than 9 months of age were more commonly affected than older foxes (p = 0.03). Plasma taurine concentrations were significantly less (p less than 0.01) in foxes that had dilated cardiomyopathy (26.8 +/- 16.4 nmol/ml) than in the control foxes (99.3 +/- 60.2 nmol/ml). A significantly higher (p less than 0.01) incidence of dilated cardiomyopathy was present in foxes with a history of a sibling or offspring that died of dilated cardiomyopathy than in foxes without a family history of cardiac death. In one fox with dilated cardiomyopathy that was tested, the myocardial taurine concentration was lower (1.7 mumol/gm wet weight) than that of control foxes (7.3 +/- 1.6 mumol/gm wet weight). Hepatic cysteinesulfinic acid decarboxylase activity was significantly less (p less than 0.001) in foxes with dilated cardiomyopathy (0.97 +/- 0.2 nmol/mm.mg protein) than in control foxes (2.11 +/- 0.07 nmol CO2/mm.mg protein).(ABSTRACT TRUNCATED AT 250 WORDS)

Severe masseter myonecrosis in a horse.

A 6-year-old Quarter Horse was examined because of acute, severely swollen masseter muscles (palpation of which elicited pain response), exophthalmos, severe chemosis, and protrusion of the third eyelids. Blood selenium and vitamin E concentrations, and results of feed analysis and muscle biopsy supported a diagnosis of nutritional myopathy. The horse was treated and was clinically normal 2 weeks after discharge from the hospital.

Local anesthetics and nuclear medical bone images of the equine fore limb.

The effects of two local anesthetic agents on the diagnostic quality of nuclear medical bone images (NMBIs) of distal parts of the equine fore limb were investigated. Local effects on bone uptake of technetium 99m methylene diphosphonate (99mTc-MDP) 4 and 24 hours after perineural and intraarticular injection of mepivacaine hydrochloride and bupivacaine hydrochloride were evaluated in the carpal and metacarpophalangeal regions of 12 horses and ponies. Neither mepivacaine hydrochloride nor bupivacaine hydrochloride significantly altered the diagnostic quality of the NMBIs. The injection and subsequent action of local anesthetics do not appear to influence local bone uptake of 99mTc-MDP significantly.

Nuclear medical bone imaging and targeted radiography for evaluation of skeletal neoplasms in 23 dogs.

Nuclear medical bone imaging was used in combination with targeted radiography to detect metastatic or multicentric lesions in 23 dogs with skeletal neoplasms. Each dog with a radiographically diagnosed skeletal neoplasm was injected with Technetium-99m labeled methylene diphosphonate (99mTc-MDP) (15.0 mCi intravenously) for whole body imaging. All areas with increased uptake of 99mTc-MDP were radiographed. In 19 dogs, the amount of bone shaft involvement in primary sites indicated by bone imaging was larger than the amount indicated by radiography. Eighteen dogs had secondary areas of increased 99mTc-MDP uptake, six of which had secondary areas that were suspected radiographically to be neoplastic. Four dogs had lesions characterized histologically as neoplasia, one as bone infarction and one as normal cortical bone. Positive predictive value for this strategy was 66.7%. Results of this study suggest that nuclear medical bone imaging with targeted radiography is a quick, noninvasive technique with a good positive predictive value for evaluation of the skeleton for metastatic or multicentric sites of neoplasia.

Influence of magnesium, water, and sodium chloride on urolithiasis in veal calves.

Forty veal calves were fed milk replacers containing differing concentrations of Mg in a 16-w study. The concentrations were .1% (basal), .3%, .6% Mg, and .6% Mg plus 2% NaCl. The four groups were further subdivided into two groups fed reconstituted milk replacer at 14 or 17.9% DM. Weight gain was greater at the two lower concentrations of Mg. Calves fed .6% Mg or .6% Mg plus NaCl excreted significantly more Mg than calves fed .1 or .3% magnesium. The low water group (17.9% DM) excreted a more concentrated urine relative to Mg, Ca, and P than did the high water group (14% DM). No urethral obstructions were observed. Seventy percent of calves at .6% Mg and 30% of calves at .6% Mg plus NaCl had stones in their kidneys consisting primarily of calcium apatite and secondarily of struvite. Added NaCl reduced kidney and bladder calculi formation. When high amounts of water were fed, added NaCl prevented calculi formation completely. All calves fed the NRC level of .1% Mg performed normally.

Reproductive patterns in cyclic and pregnant thyroidectomized mares.

Three Quarter-horse mares were thyroidectomized at about 1.5 years of age. Three similar intact mares served as controls. The study continued through two breeding seasons. The thyroidectomized mares were lethargic, rear limbs were oedematous and hair coats were coarse. They displayed a tranquil oestrous behaviour when exposed to a stallion and were only mildly antagonistic when not in oestrus. Length of oestrous cycles varied but most often they were 19-24 days long. Duration of oestrus (mean +/- s.e.m.) for the control and thyroidectomized mares was 12.9 +/- 2.9 and 11.7 +/- 2.2 days respectively (P greater than 0.05). The peak of LH during oestrus was as high as 60 ng/ml blood serum with no difference between the two groups. Peak progestagen on Day 7 after ovulation for controls was 9.0 +/- 1.6 ng/ml and was not different (P greater than 0.05) from the peak of 6.3 +/- 1.7 ng/ml for thyroidectomized mares on Day 8. Pregnancy was achieved in both groups of mares, including the use of semen from a thyroidectomized stallion. Thyroxine was detectable in one pregnant thyroidectomized mare during the last two-thirds of pregnancy only.

Scintigraphic diagnosis of primary hepatocellular carcinoma in the woodchuck (Marmota monax).

Hepatic imaging with 99mTc-sulfur colloid was used to diagnose primary hepatocellular carcinoma (PHC) in woodchucks infected with woodchuck hepatitis virus (WHV). Based on imaging results, 6 of 12 WHV-infected woodchucks had space-occupying hepatic lesions, and all 6 had PHC. Of the remaining woodchucks, 2 did not have PHC, 2 had discrete tumors (less than 1 cm diameter), 1 had miliary small tumors, and 1 had tumors located near the great vessels. Hepatic imaging was a valuable technique for diagnosis of PHC in WHV-infected woodchucks.

Familial nonspherocytic hemolytic anemia in poodles.

Nonspherocytic hemolytic anemia, characterized by marked reticulocytosis, hepatosplenomegaly, hemosiderosis of reticuloendothelial organs and bone marrow myelofibrosis, and osteosclerosis, was diagnosed in 5 related Poodles. The unremitting anemia was clinically evident by 1 year of age, and was fatal as early as 3 years of age. Despite intense diagnostic endeavors including RBC fragility studies, RBC enzyme assays, and hemoglobin electrophoresis, the cause of this nonspherocytic hemolytic anemia remains to be determined.

Repair of a delayed-union stress fracture of the dorsal cortex of a metacarpal bone in a horse.

A delayed union metacarpal stress fracture was repaired in a horse, using lag-screw fixation in conjunction with dc electrical stimulation. Twelve weeks after surgery, radiographic and scintigraphic evaluations revealed that the fracture line was not discernible and that there was a decrease in radiopharmaceutic uptake, as compared with that in previous bone imaging studies. In this horse, DC stimulation of a delayed union stress fracture in a bone resulted in a healing pattern similar to that in human beings with delayed union fractures when treated with electrical stimulation.

Capacitively coupled electrical stimulation of bone healing in the horse: in vivo study with a Salter type IV osteotomy model with stainless steel surface electrodes.

The use of capacitively coupled low-voltage signals for stimulation of osteogenesis has been reported in a variety of animal models. Electrically induced osteogenesis was investigated with a capacitively coupled electric field on a radius (distal-lateral orientation) osteotomy model, in conjunction with internal fixation and postoperative loading. Twelve adult horses of either sex were allotted to 2 groups of 6; 1 group was given electrical stimulation and the other served as controls. A low-voltage high-frequency capacitively coupled electrical signal was locally and continuously applied to the electrically stimulated group for 60 days through external, bare stainless steel surface electrodes which were placed on the skin in circuit with a small, portable power source. Harness compatibility and stimulator and battery durability were excellent. However, stainless steel electrodes required a rigid maintenance schedule to maintain consistent current levels. Synovial fluid evaluation demonstrated intra-articular inflammation (decreased viscosity, hyaluronic acid, and increased protein concentration) 1 week postoperatively that generally improved during subsequent weeks and no distinction between groups was observed at 60 days. Radiographically, there was no difference in the appearance of the healing process of control and that of stimulated horses during the 60 days. Angiography showed bridging blood vessels in both groups. Uptake of a bone seeking radiopharmaceutical peaked at 3 weeks in both groups and was 1.92 +/- 0.6 cps/pixel/mCi and 1.26 +/- 0.40 csp/pixel/mCi for control and stimulated horses, respectively. At any given observation period, uptake in the lesion area was greater in the control group. Ultimate strengths of trabecular bone in 60-day control radii and stimulated radii were 12.64 +/- 3.013 and 9.60 +/- 3.95 MN/m2, and the flexural moduli of elasticity were 698.0 +/- 423 and 402.0 +/- 523 MN/m2, respectively. Porosity index was similar for all specimens. Gross, histologic, and microradiographic evaluations indicated that controls healed more efficiently than stimulated horses. A capacitively coupled applied voltage of 2.2 V RMS (mean) producing a current of 17.32 mA (mean) did not stimulate sufficient bone production in a metaphyseal osteotomy model to affect the mechanical properties of the bone or accelerate the healing process.

Some nutritional problems of horses.

The effects of overfeeding, calcium-phosphorus imbalance, misuse of supplements and false advertising on equine nutrition are discussed. Overfeeding is known to cause disorders in several species but, although a similar relationship has been suggested on clinical evidence, no controlled trials on horses have been reported. It has also been suggested that overfeeding is a problem only for those horses with a genetic predisposition to skeletal problems. The importance of adequate calcium and phosphorus levels has been known for many years but severe cases of calcium deficiency still occur. Client education is important and should not be neglected. Excessive use of supplements containing high levels of trace minerals (eg, iodine and selenium) or fat soluble vitamins (eg, vitamin A and vitamin D) can be harmful. Some manufacturers advertise supplements in terms which may inadvertently or intentionally misrepresent their products. Supplements should, therefore, be selected carefully to ensure that they meet the particular requirements of the individual.

Radionuclide joint imaging: an ancillary technique in the diagnosis of canine hip dysplasia.

Radionuclide joint imaging has been adapted for use in the diagnosis of canine hip dysplasia. Studies were done on 29 growing Labrador Retriever pups between 3 months and 1.5 years of age and on two adult dogs. Initally, pelvic radiographs and joint images were done on pups 12 to 16 weeks of age and were repeated at 2-week intervals thereafter. 99mTechnetium methylene diphosphonate (99mTcMDP), an osteotrphic agent, was the radiopharmaceutical used for all imaging studies. Inteference due to 99mTcMDP accumulation in the urinary bladder was avoided by static abdominal pressure. In 19 of the 21 dogs identified as abnormal by pelvic radiography, a similar diagnosis based on joint imaging was made for a true positive ratio of 0.90. Eight of ten dogs were identified as normal by means of both procedures, for a true negative ratio of 0.80. The overall accuracy was 0.87. Joint imaging was thus shown to be of similar value as pelvic radiography in the diagnosis of hip dysplasia. Joint images, however, displayed information on bone metabolic activity that could not be determined by pelvic radiography.

Ascorbic acid deficiency and hypertrophic osteodystrophy in the dog: a rebuttal.

Plasma ascorbic acid (PAA) in normal Labrador Retriever dogs less than one year of age averaged 1.22 +/- 0.05 mg/dl (x +/- sem) and was significantly higher than the value of 0.89 +/- 0.03, for Labrador Retrievers two years of age and older. No significant diurnal variation in PAA was observed. Oral or intravenous administration of 0.5 or 1.0 g of ascorbic acid (AA) elevated PAA for less than 8 hours. Injection of ACTH caused a significant decline in PAA for the initial 2 days, with variable results thereafter. Labrador Retriever puppies fed a ration high in protein, energy and calcium developed the typical skeletal diseases of overnutrition, including hypertrophic osteodystrophy (HOD). The addition or oral AA (0.5 g twice daily) had no ameliorating effect on the skeletal lesions. Instead AA supplementation resulted in relatively higher serum calcium values which, presumably by enhanced hypercalcitoninism, decreased bone resorption. Thus, AA treatment of dogs with HOD is contraindicated, as it can only aggravate the osseous lesions of HOD. The decreased PAA reported in dogs with HOD is interpreted to be the result of stress from pain.