Alkaline phosphatase and its isoenzymes in the tissues and sera of normal dogs

The total alkaline phosphatase (AP) activity and the pattern of its isoenzymes were studied in the tissues and sera of normal adult dogs. Small intestine mucosa showed the greatest total AP activity followed by kidney, bone, pancreas, liver, lung, skeletal muscle and heart muscle. After separation by agarose gel electrophoresis, each tissue showed only one isoenzyme except lung which showed two. The tissue isoenzymes, in decreasing order of migration distance towards the anode, were as follows: fast lung isoenzyme, liver or slow lung isoenzyme, the group consisting of skeletal muscle, bone, small intestine and pancreas isoenzymes and, finally, the kidney isoenzyme. Two isoenzymes occurred in serum. The major band corresponded to liver and the slow lung isoenzyme, while the minor band was considered to be the corticosteroid-induced isoenzyme, previously thought to be absent from normal serum. The AP isoenzyme patterns in lung and skeletal muscle and the presence of an isoenzyme migrating an identical distance to the corticosteroid-induced isoenzyme do not appear to have been reported before in normal dogs.     

Alkaline phosphatase and its isoenzymes in the tissues and sera of normal dogs

The total alkaline phosphatase (AP) activity and the pattern of its isoenzymes were studied in the tissues and sera of normal adult dogs. Small intestine mucosa showed the greatest total AP activity followed by kidney, bone, pancreas, liver, lung, skeletal muscle and heart muscle. After separation by agarose gel electrophoresis, each tissue showed only one isoenzyme except lung which showed two. The tissue isoenzymes, in decreasing order of migration distance towards the anode, were as follows: fast lung isoenzyme, liver or slow lung isoenzyme, the group consisting of skeletal muscle, bone, small intestine and pancreas isoenzymes and, finally, the kidney isoenzyme. Two isoenzymes occurred in serum. The major band corresponded to liver and the slow lung isoenzyme, while the minor band was considered to be the corticosteroid-induced isoenzyme, previously thought to be absent from normal serum.The AP isoenzyme patterns in lung and skeletal muscle and the presence of an isoenzyme migrating an identical distance to the corticosteroid-induced isoenzyme do not appear to have been reported before in normal dogs.     

Lactate dehydrogenase and its isoenzymes in the tissues and sera of clinically normal dogs

The total activity of lactate dehydrogenase (LDH) and the percentage distribution of its isoenzymes in the tissues and sera of clinically normal adult dogs are presented. Total LDH activity was greatest in skeletal muscle followed by heart muscle, kidney, small intestinal mucosa, liver, lung, pancreas and bone. Each tissue had a unique isoenzyme pattern and the proportions of the isoenzymes in serum suggested that liver is the source of normal serum LDH. The tissue isoenzyme patterns were similar to those obtained by other authors in human beings, horses, cattle, sheep and cats although in liver, differences between ruminants and monogastric animals including dogs were evident. The data presented provide a basis for the interpretation of serum LDH isoenzyme patterns in canine disease.     

Gamma-glutamyl transpeptidase and its multiple forms in the tissues and sera of normal dogs

The total gamma-glutamyl transpeptidase activity and the proportions of its multiple forms in canine tissues and serum have been measured. The kidney showed the greatest activity, followed by the pancreas and small intestine. After separation by cellulose acetate electrophoresis, two main areas of activity were observed in tissues, an insoluble fraction which remained at the origin and a soluble fraction which migrated towards the anode. The soluble fraction was divisible into six zones differing in mobility and, with the exception of liver and lung, the tissues differed in the distances migrated by these soluble fractions. Serum gamma glutamyl transpeptidase formed one zone corresponding to the soluble fraction of liver and lung. The separation of the multiple forms is of potential value in determining the source of increased serum activities of the enzyme in the dog.     

ELISA to determine anti-K99 pilus antibody in the sera of normal and diarrhoeic calves

An ELISA to detect circulating antibodies against K99 pili, a major attachment factor to intestinal epithelial cells of Escherichia coli in calves, was performed. Two methods of K99 pili purification were attempted. Best results in terms of purity of the K99 antigen were achieved following the method described by Karkhanis and Bhogal (1986). This procedure included a heat shock at 65°C during 25 min to release the pili and ultracentrifugation steps to purify the antigen. SDS-PAGE showed an 18 KDa major band, identified as the K99 pilus antigen after immunoblotting against reference antisera. The purified K99 antigen was then adsorbed to the ELISA microplates. High optical density was obtained in the ELISA using a pool of sera from immunized cows. No differences in antibody levels (P ≥ 0.05) could be detected between clinically healthy calves and those showing diarrhoea.     

Antibodies against Helicobacter felis in sera of cats and dogs.

Serum samples from 61 dogs and 49 cats were screened for circulating antibodies against Helicobacter felis by an enzyme-linked immunosorbent assay (ELISA) using sonicated bacteria as an antigen. To improve the specificity of the ELISA, sera were absorbed with Campylobacter jejuni subsp. jejuni H. pylori as well as H. felis. Sera from 26 dogs (43%) and 19 cats (39%) revealed clear positive absorbance readings determined as an optical density (OD) that was statistically significant above the OD mean value [P < 0.025 (one-tailed); log10]. The absorbance pattern of ELISA-positive sera corresponded to results obtained with bovine and human reference sera. Furthermore, a correlation between the immune response and results from histopathological examination of gastric specimens from 22 dogs was demonstrated. It could be shown that antibodies against H. felis in sera of cats and dogs can easily be detected using an ELISA. The diagnostic value of this test must be evaluated in further investigations.     

Electrophoretic evaluation of sera from dogs with cancer.

A quantitative electrophoretic method was developed for evaluating sera from dogs with naturally occurring malignant tumors. Electrophoretic migration ratios (EMR) were devised to standardize characteristic protein components and to analyze alterations in mobility of these protein components, using a computerized densitometric technique. Sera values from dogs with lymphosarcomas, osteosarcomas, and mammary tumors were compared, using conventional methods of statistical analysis. Results indicate a statistical difference existed between mean values of EMR and alpha2-globulins of sera from dogs with tumors and those values from normal dogs. In addition, unique EMR and concentration of protein components were identified for dogs with lymphosarcomas, osteosarcomas, and mammary tumors.     

Detection of anti-Babesia gibsoni heat shock protein 70 antibody and anti-canine heat shock protein 70 antibody in sera from Babesia gibsoni-infected dogs

Antibodies that recognized either Babesia gibsoni or canine red blood cell (RBC) 70-kilodalton (kDa) protein were detected in serum from acutely and chronically B. gibsoni-infected. In those sera, antibodies that reacted with recombinant B. gibsoni and canine heat shock protein 70 (rBgHsp70 and rcHsp70) were detected; therefore, B. gibsoni and canine RBC 70-kDa proteins seemed to be BgHsp70 and cHsp70, respectively. In infected dogs, the amounts of these antibodies increased after infection. Interestingly, polyclonal antibody raised against rBgHsp70 in two rabbits reacted not only with rBgHsp70 but also with rcHsp70 and native cHsp70 from canine RBCs. Because BgHsp70 showed high homology with cHsp70 (70.8%), anti-rBgHsp70 antibody might cross-react with cHsp70. Additionally, the localizations of both BgHsp70 and cHsp70 were observed by indirect fluorescence assay. As a result, cHsp70 was not found on the membrane surface of erythrocytes, suggesting that erythrocytes would not be targets of anti-cHsp70 antibody. Meanwhile, only exoerythrocytic parasites were stained by anti-rBgHsp70 antibody. This result showed that BgHsp70 would be expressed on the surface of parasites during the exoerythrocytic stage. These results indicated that BgHsp70 was a highly immunogenic protein in canine B. gibsoni infection, and that exoerythrocytic parasites might be targets of anti-BgHsp70 antibody.     

Lipids and lipoproteins in normal dogs and in dogs with secondary hyperlipoproteinemia.

Concentrations of serum cholesterol, triglyceride, and free glycerol were neasured, and serum lipoprotein electrophoresis was performed in normal dogs and in dogs with hypothyroidism, diabetes mellitus, and acute pancreatitis. Twelve privately owned dogs and 20 Basset Hounds from a research colony served as normal subjects. Privately owned had higher serum lipid concentrations than did research dogs. On electrophorisis of normal dog serum, lipoprotein bands were detected at the beta, pre-beta, alpha-2, and alpha-1 positions. Hypercholesterolemia was associated with increased intensity of the alpha-2-lipoprotein band, and hypertriglyceridemia occurred with increased lipoprotein at the origin, or the beta or pre-beta positions. Hypothyroid dogs had normal lipid values and lipoprotein electrophoretic patterns, hypercholesterolemia with increased intensity of the alpha-2-lipoprotein band, or hypercholesterolemia and hypertiglyceridemia with prominent beta-, pre-beta, and alpha-2-lipoprotein electrophoretic patterns were changed to near-normal values after levothyroxine administration. Diabetic dogs had increased serum cholesterol and triglyceride content and a predominance of lower density lipoproteins, as detected by electrophoresis. Insulin therapy caused the lipid concentrations to decrease and the electrophoretic pattern to revert to near normal. Dogs with acute pancreatitis had moderately increased serum lipid content and electrophoretic patterns characterized by increased intensity of the beta-lipoprotein band and by altered migration of alpha-1-lipoproteins.     

Detection of adenovirus precipitating antibodies in the sera of Polo horses in Nigeria.

Serum samples obtained from 107 Polo horses showing clinical signs of viral respiratory disease were tested for precipitating antibodies to adenovirus by agar gel precipitation test and counter-immunoelectrophoresis method. The results obtained demonstrate serological evidence of adenovirus infection in Polo horses in Nigeria. The counter-immunoelectrophoresis method was observed to be about 3 times more sensitive than the agar gel precipitation test with 19.3 vs 64.5%. It could thus be used to screen a large number of serum samples within a short period.     

Immunoperoxidase study of adenovirus pneumonia in dogs

Summary

The pathology of adenovirus pneumonia in 16 dogs is described. Clinically, these dogs had been severely ill, with severe dyspnoea and listlessness, but only faint coughing.

Histopathological lesions could be associated directly with the presence of adenovirus antigens in the lungs of these dogs by using an unlabelled immunoperoxidase technique on paraffin tissue sections. The lesions were focal and located in alveoli and bronchioles. Infected cells were mostly alveolar macrophages and less frequently type I and 2 pneumocytes and bronchiolar epithelial cells. Infiltrating neutrophils and lymphocytes were not observed to be infected.

This type of pneumonia appears to be a fairly well defined clinical and pathological entity in kennel dogs.     

Plasma adrenocorticotropin levels in normal dogs.

Plasma concentrations of adrenocorticotropin (ACTH; corticotropin) were measured in 31 normal dogs (house pets) at rest. The mean concentration was 45.77 pg/ml of blood, with individual values ranging from 17 to 98 pg/ml of blood. The measurement of this hormone secreted by the anterior lobe of the pituitary gland (adenohypophysis) may aid in the diagnosis of spontaneous canine adrenal cortical disorders, both primary and secondary.     

Tumour-associated antigens in bovine ocular squamous cell carcinoma: studies with sera from tumour-bearing animals

Sera from 21 cattle (10 bovine ocular squamous cell carcinoma (BOSCC) bearing and 11 controls) were tested for antibody reactivity against various cultured cells (BOSCC, normal skin and tumours other than BOSCC), by an indirect immunofluorescence technique. Most of the BOSCC sera reacted with autologous and allogeneic BOSCC cells and did not show any significant reactivity with normal skin cells (autologous or allogeneic). These sera when further tested on 7 different allogeneic or xenogeneic tumour cell types (other than BOSCC), showed significant reactivity only with cultured equine sarcoid and cutaneous papilloma cells. Three of the BOSCC sera did not react with autologous BOSCC cells but reacted indiscriminately with all other allogeneic BOSCC, normal cells and other tumour cells. Most of the control sera did not show any significant reactivity against BOSCC cells except sera from one cow bearing ocular papilloma and 2 healthy normals that were in contact with BOSCC bearing animals. This study suggests that BOSCC cells possess tumour associated antigens that are common to most (if not all) BOSCC cells.     

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).     

Encephalomyocarditis virus antibodies in sera from apparently normal pigs.

Antibodies which neutralise and precipitate encephalomyocarditis virus habe been found in the serum of more than 28 per cent of normal pigs in Britain. Neutralising activity was found in association with several size classes of antibody.     

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.