Serum isoamylases and canine pancreatic insufficiency.
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Biomedical subjects
Publications and source records attributed to D L Doxey.
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In human medicine it has been shown that the bromocresol green (BCG) dye-binding method for the determination of serum albumin is not entirely specific, the dye reacting also with certain human serum globulins. This causes over-estimation of albumin when reaction times are prolonged beyond 30 seconds. In the present study, serum albumin values obtained from three animal species by the "immediate", i.e. less than 30 seconds, BCG reaction were compared with those by the 10-minute BCG reaction. Albumin-depleted sera were prepared using an affinity chromatography technique and their reactions and those of purified gamma globulin preparations with the dye were studied. In cattle, sheep and horses, serum albumin values obtained by the 10-minute reaction were higher than those obtained by the "immediate" BCG reaction, the differences being statistically significant. Purified gamma globulin did not react with the BCG dye after 10 minutes, but other globulins did. There were differences between the species in the magnitude of the reaction of their globulins with BCG dye.
The effect of age, sex and growth rate on total serum lactate dehydrogenase (LDH) and its isoenzymes was assessed in Suffolk X Halfbred lambs from 1.5 days to 10 weeks old. The total LDH activity was higher at 1.5 days than in older lambs and the percentage of LDH1 tended to increase while LDH4 and LDH5 decreased with age from 2 weeks, indicating that the isoenzyme distribution was changing towards the adult pattern. In female lambs, the percentage of LDH1 was higher than in male lambs at 1.5 days, but little importance was attached to this finding since the difference was not significant at 10 weeks. No clear relationships existed between the level of total LDH or its isoenzymes and daily liveweight gain. The age-related changes were considered to be of greater significance than those related to sex and growth rate when interpreting serum LDH levels in lambs.
Total lactate dehydrogenase and the absolute and percentage levels of its isoenzymes were measured in lung lesions and macroscopically normal areas of lung from lambs with chronic proliferative exudative pneumonia and acute pasteurella pneumonia. Lung lesions had a higher total enzyme activity which was associated mainly with increases in the activity of the LDH4 and LDH5 isoenzymes, particularly in chronic pneumonia, and gave lung lesions a considerable potential for altering the serum isoenzyme distribution. Thus, the nature of any changes in the serum isoenzyme distribution will depend on whether the isoenzymes are released from abnormal or normal areas of lung. This appears to be the first report on lactate dehydrogenase isoenzymes in ovine pneumonia.
Estimations were made of the serum isoamylase values of normal dogs and of dogs with confirmed exocrine pancreatic insufficiency. A statistically significant difference was demonstrated between the two groups in respect of the values of one of the isoamylase fractions measured. Further study has confirmed that canine salivary tissue lacks amylase activity and that the source of the isoamylase fractions was the pancreas. This knowledge has potential value in the diagnosis of canine exocrine pancreatic insufficiency.
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Lactate dehydrogenase in ovine tissues was separated by electrophoresis, one dimensional isoelectric focusing (IEF) and a two dimensional technique. Tissues showed five zones of enzyme activity consisting of multiple bands after IEF. The IEF zymograms were unique for each tissue and differed from those of other species. Two dimensional separation revealed that the five zones of activity observed on IEF corresponded to the five isoenzymes separable by conventional electrophoresis.
Two types of glucosephosphate isomerase (GPI) are described for the first time in sheep. Type T produced a 3-band pattern on polyacrylamide gel electrophoresis of serum samples, whereas type M produced a pattern of 5-7 bands. When 23 Scottish Halfbred ewes showing type T were mated to 3 Suffolk rams showing type M, their 32 offspring had frequencies of 0.72 and 0.28 for T and M respectively. There was no conclusive evidence that the types were controlled by codominant alleles or by simple Mendelian inheritance and neither was sex-linked. The frequency of the rarer type (M) was sufficiently high to provide conclusive evidence of true enzyme polymorphism.
Serum albumin concentrations were measured by the immediate reacting bromocresol green (BCG) method and by agarose gel electrophoresis in healthy cattle, sheep and horses. No statistically significant differences were found between the values obtained by the two methods. The immediate reacting BCG method is quicker and cheaper when used under the conditions described.
A new method of quantifying fat absorption based on the estimation of serum triglyceride levels is described. When long chain triglyceride (LCT) was fed to normal dogs a significant elevation of serum triglyceride concentration was recorded which was not observed in dogs with exocrine pancreatic insufficiency. When the test was repeated on the dogs with pancreatic insufficiency using the LCT together with lipase the serum triglyceride concentration increased, suggesting the failure in the initial test was caused by a deficiency in pancreatic lipase. Feeding medium chain triglyceride (MCT) did not raise the serum triglyceride levels in normal dogs.
The normal activities of lactate dehydrogenase and creatine kinase isoenzymes in the tissues and serum of clinically normal five-and-a-half-month-old lambs are presented. The percentage distribution of the lactate dehydrogenase isoenzymes in serum, liver, heart, lung and skeletal muscle were in agreement with previous studies but the distribution in ovine abomasum, small intestine, large intestine and red blood cells has not been previously described. Up to five creatine kinase isoenzymes were detected in ovine tissues and four in serum. One creatine kinase isoenzyme present in serum and tissues was thought to represent a mitochondrial isoenzyme which is absent from the normal serum of other species. Estimations of both lactate dehydrogenase and creatine kinase isoenzymes allowed all the tissues examined to be distinguished and were therefore more likely to allow tissue-specific isoenzyme patterns to be detected in serum than the estimation of the isoenzymes of either enzyme alone.
Serum samples from five species of domestic animals were subjected to protein electrophoresis on commercially prepared agarose gel films. After processing, the strained electrophoretic patterns and densitometric traces exhibited certain characteristics for each species. The features which were considered distinctive of each species are illustrated.
Samples of serum from both young and adult normal cattle, sheep and horses were subjected to protein electrophoresis on agarose gel films. After processing, the stained electrophoretic patterns and densitometric traces of each species exhibited certain specific characteristics. The separations also revealed differences between young and adult animals of the same species. These age-related differences are described here.
Studies involving 49 ewes which were well nourished during the last eight weeks of pregnancy indicated that ewe weight loss during pregnancy and lamb birth weights followed the recognised pattern and were within acceptable "normal" limits. Colostrum production ranged from 1238 to 4593 g per ewe during the 48 h following the first suckling and there was evidence that production levels were related to demand by the lambs. Colostrum consumption by individual lambs was related to litter size. The smaller the litter size the greater was the amount ingested during the first 48 h of life. No clear relationship was established between the amount of colostrum consumed and the level of immunoglobulin in a lamb's circulation. Possible reasons for this are discussed.
The immunoglobulin content of ewe colostrum falls rapidly after the first suckling and reaches a low level 36 h after the first feed. The whey content of colostrum will rise by more than 50 per cent during the same period. The production of colostral immunoglobulin during the 24 h following the first feed was related, in this work, to the amount of colostrum produced; the greater the colostrum production, the greater the immunoglobulin production. Colostrum production ranged from 1216 to 4493 g and immunoglobulin production from 22-21 to 86-34 g during the eight feeds studied. Colostrum production was related to demand by the lambs. There was a clear positive correlation between the total amount of immunoglobulin the lambs' circulation 30 h after the first feed and the immunoglobulin consumed. Approximately 20 to 25 per cent of the immunoglobulin ingested was present in the lambs' circulation at this time.
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A total of 351 single-suckled beef calves were blood sampled at twice-weekly intervals for the first two and a half weeks of life. Twenty three of them died, 13 of a syndrome characterised by acute collapse and 10 of diarrhoea which had persisted for several days before death. Those which died acutely showed a sudden terminal rise in blood levels of potassium, magnesium, inorganic phosphate and total protein. Those which died after several days of diarrhoea showed a more gradual increase in blood chloride and urea concentrations and in packed cell volume values. It is suggested that these changes indicate a difference in the pathogenesis of the two situations. Calves which died had lower blood glucose levels before the onset of clinical signs than those which survived. It is suggested that this may have been a contributory factor in their mortality.
An experiment was conducted to investigate the mechanisms responsible for the brain lesions, diarrhoea and haemorrhages produced by amprolium poisoning of preruminant lambs. The encephalopathy was preceded by a reduction of cerebrocortical transketolase activity. Diarrhoea was not associated with histological evidence of pathological change in the small intestine, and in lambs with severe diarrhoea the small intestinal levels of alkaline phosphatase and lactase were unaffected. Haemorrhages were associated with a thrombocytopenia which was attributed to degeneration of bone marrow megakaryocytes. The bone marrow of the majority of the lambs which had received amprolium for 3 wk or more was severely depopulated, erythrocyte precursors being the most severely affected. The marrow depopulation was atrributed to decreased cell production, as the majority of the remaining cells showed little evidence of degeneration and the number of mitotic figures in the marrow of amprolium-treated lambs was considerably reduced as compared with the controls.