Value of measurements of the serum activities of creatine phosphokinase, aspartate aminotransferase and lactate dehydrogenase for predicting whether recumbent dairy cows will recover.

Over a period of seven years, 262 recumbent dairy cows were investigated, and serum samples were analysed for creatine phosphokinase (CPK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) activities. The results from cows for which all three measurements were available, were analysed statistically by the non-parametric receiver operating characteristic (ROC) method, to evaluate the value of the serum enzyme activities for predicting a failure to recover. The sensitivity and specificity of the activity levels were calculated over all cut-off points, and ROC curves were created by plotting sensitivity as a function of 1--specificity at each cut-off point for samples grouped by the interval after the cows became recumbent. The predictive values of the tests were compared by calculating the areas under the curves, and the positive and negative predictive values of the tests were calculated and plotted against the prevalence of a failure to recover. The results from each of the enzymes were significantly better than from a random test with no discriminatory ability. The optimal cut-off points maximising the sensitivity and specificity of the tests were 2330, 2225 and 171 U/litre for CPK, LDH and AST, respectively. The predictive value of AST was significantly better than that of CPK or LDH, and measures made on the second and third day of recumbency were significantly better with optimal cut-off points of 128 and 189 U/litre, respectively.

Evaluation of buffalo colostrum quality by estimation of enzyme activity levels.

A study to evaluate the value and potential use of colostral enzymes as markers for the evaluation of buffalo colostrum quality was conducted. The enzymes gamma-glutamyltransferase (GGT), lactic dehydrogenase (LDH), and alkaline phosphatase (ALP) in buffalo's colostrum were measured spectrophotometrically, and their activities were correlated with the gamma-globulin content. Gamma-globulin concentration was determined following the electrophoretic separation of the colostral proteins and quantified with a densitometer. Colostrum was obtained from 15 dams, soon after calving. Means, standard deviations, correlation coefficients, and degree of significance were calculated using the general linear model procedure of the Statistical Analysis Systems program. The activity of GGT in the colostrum was the highest, followed by LDH and ALP. A significant correlation (r = 0.86; P < 0.001) was seen between GGT and gamma-globulin concentration in the colostrum, supporting the suggestion of using this enzyme as a marker for the evaluation of colostrum quality.

Buffalo-milk enzyme levels, their sensitivity to heat inactivation, and their possible use as markers for pasteurization.

The activities and rates of inactivation of four enzymes in raw buffalo milk were measured in relation to the process of heating to determine the value of these enzymes as markers for the evaluation of milk pasteurization. The activities of the enzymes alkaline phosphatase (ALP), lactic dehydrogenase (LDH), gamma-glutamyltransferase (GGT), and aspartate aminotransferase (AST) were measured before and after heating at 50, 60, 70, and 80 degrees C for 1, 3, 5, 10, 20, and 30 min. The enzyme GGT showed the highest activity (712 +/- 601 IU/liter), followed by LDH (386 +/- 183 IU/liter), ALP (295 +/- 164 IU/liter), and AST (18 +/- 4 IU/liter). Heating the milk at 50 degrees C for 1 to 30 min resulted in no effect on the activity of any of the enzymes. At 60 degrees C, ALP showed the highest sensitivity to heat inactivation, whereas all other enzymes showed resistance. At 70 degrees C, ALP activity was abolished completely after 1 min, whereas GGT and LDH lost most activity after 10 min, and AST still maintained 50% activity even after 30 min. At 80 degrees C, the activities of LDH and GGT were lost, whereas AST still retained some of its activity. The results suggest that in addition to ALP, LDH and GGT, but not AST, are potential markers for heat denaturation in buffalo milk, with GGT having the advantage that its concentration is the highest.

Biochemical alterations secondary to disuse atrophy in the rat's serum and limb tissues.

Disuse atrophy has been the subject of research studies of an animal model in which single-limb immobilization induces atrophic changes in the immobilized limb. These reveal systemic changes in the experimental animals that go far beyond the local response expected in that situation and are not fully understood as yet. We therefore performed a biochemical study on the effect of hind-limb immobilization on the serum and tissues of rats. The experiment was carried out on 70 young Sprague-Dawley male rats. In one group of 35 rats, the left hind-limb was immobilized for 3 weeks. Another group of 35 rats served as controls. Serum total protein, albumin, urea, creatinine, and calcium were found to be reduced during immobilization. Serum cholesterol levels, on the other hand, increased to a considerable extent. No changes were recorded with phosphate, bilirubin, and magnesium. Serum alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were both reduced in activity. The activity of muscle aspartate aminotransferase (AST) and bone alkaline phosphatase (ALP) was also decreased. Lactate dehydrogenase (LDH) remained unchanged in both serum and muscle. We discuss our findings in the light of previous knowledge regarding the atrophic process.

The effect of single hind-limb immobilization on the contralateral limb in the rat: a morphometric and biochemical study.

Disuse atrophy has been the subject of research studies that make the use of an animal model in which single-limb immobilization induces atrophic changes in the immobilized limb. The effect upon the nonimmobilized contralateral limb is, however, still unclear, and findings in the literature are inconclusive. We therefore performed a multidisciplinary study to clear this issue. The experimental population was 70 young male Sprague-Dawley rats. In one group of 35 rats, the left hind limb was immobilized for 3 weeks. Another group of 35 rats served as controls. limb and tibial bone weights (both "dry" and "ash") were found to be reduced in both hind limbs. Bone alkaline phosphatase (ALP) activity was likewise reduced in both limbs. We conclude that the contralateral hind limb in the rat is clearly subjected to atrophic changes that are similar in magnitude or severity to those experienced by the immobilized limb. We further discuss these findings in view of previous knowledge regarding the atrophic process.

Lactate dehydrogenase isoenzyme distribution and patterns in chicken organs.

Unlike most mammals, chicken lactate dehydrogenase isoenzymes cannot be separated using the 'Titan-Gel' electrophoresis. However, using isoelectric focusing at a pH range of 3.0 to 9.0, a good and clear separation of all five isoenzymes was achieved. Generally, three characteristic groups were seen: (a) those having a cathodic domination (breast muscle and serum) with mainly lactate dehydrogenase-5 (b) those having an anodic domination (heart, muscle, liver, pancreas, kidney, erythrocytes) of mainly lactate dehydrogenase - 1 and 2 and (c) those with a more uniform distribution (spleen, lung, and brain). The total lactate dehydrogenase activity was the highest in the breast muscle, followed by the heart muscle, liver and serum with the lowest activities in the lung and pancreas.

Laminitis and dermatitis in heifers associated with excessive carbohydrate intake: skin lesions and biochemical findings.

The effects of a sudden addition of a large quantity of readily fermentable carbohydrate to the feed ration of pregnant heifers are described. Clinical and pathological changes caused by the resulting disease were confined to the digits and skin. The 4 acutely affected heifers were reluctant to get up or move (group II). They tended to lie down or stand with feet bunched together and the back arched, often shifting weight from limb to limb. They walked stiffly with great tenderness and pain in the digits. Extreme pain was noticed when the digits were examined. In 4 of 8 heifers, separation of the sole at the heel, with leakage of exudate, and under-running of the sole were observed. Necrotic dermatitis of the legs, alopecia and hyperkeratosis of the tail were noticed in all 8 heifers. Skin lesions appeared simultaneously. Four of the heifers (group I) recovered, and the other 4 (group II) were sent to slaughter. No post mortem examination was performed. The biochemical findings revealed a significantly higher concentration of total serum globulins and sodium, and increased activity, in CK, LDH and AST. A significantly decreasing pattern was noted in blood urea concentration, cholesterol, triglycerides, albumin and calcium. No significant differences among the various groups were found in the activities of amylase, GGT, and concentration of creatinine, total bilirubin, inorganic phosphorus, magnesium and potassium.

Microscopic alterations in the physis of long bones in response to hind limb immobilization in the rat.

BACKGROUND: The growth plate increases its activity in response to exercise. Likewise, decreased physical activity exerts a negative effect on bone growth and development, leading to rarefaction of the subepiphyseal bone. Limb immobilization inhibits the growth plate's activity, indirectly shown by a recorded arrest in longitudinal growth of the long bones. However, there is no direct evidence concerning the growth plate itself. OBJECTIVE: To determine whether the growth plate exhibits measurable microstructural changes in response to decreased levels of physical activity. METHODS: Histomorphometric analysis was used to qualitatively and quantitatively assess the changes in the epiphyseal plate in response to single hind limb immobilization in the rat. In 16 of 25 Sprague-Dawley male rats the left hind limb was immobilized for 3 weeks; the remaining 9 rats served as controls. The left proximal tibia of each animal was examined by computerized image analysis. RESULTS: There was a decrease in epiphyseal height, cell column density and subepiphyseal trabecular area--all indices of growth plate activity. Metaphyseal cortical thickness was also depressed, thereby confirming the efficacy of the immobilization method applied. CONCLUSIONS: Limb immobilization in the rat induces inhibitory histological changes in the epiphyseal growth plate, which are in contrast to the excitatory microscopic changes seen with exercise. These changes can be assessed quantitatively. Their potential for reversibility remains to be determined by future experiments.

Metabolic and productive responses of dairy cows to increased ion supplementation at early lactation in warm weather.

We found previously that the current recommendations for Na+, K+, and Cl- contents in the diet do not meet the needs of lactating cows. The response of cows receiving a ration with increased amounts of Na+, K+, and Cl- (E cows) were compared with those of cows consuming the same ration with a fixed concentration of these ions (C cows) between weeks 2 and 8 post partum (PP). Milk, protein, fat and lactose yields, and dry matter intake between weeks 2 and 4 PP were higher in E than in C cows. These differences did not occur between weeks 4 and 8 PP, mainly because of a higher incidence of PP complications in E cows. A greater increase in plasma insulin-like growth factor-1 concentration in E than in C animals during weeks 2 and 3 PP was consistent with the milk responses. A reduction in aldosterone concentration in E cows in weeks 2 and 3 PP was a consequence of their Na+ requirements being satisfied as a result of their enhanced Na+ intake. A subsequent elevation in aldosterone concentration in E animals was probably related to a moderate excess in K+ intake. This increase in aldosterone explains the urinary potassium loss that was detected at week 6 PP. The absence of differences between E and C cows in plasma renin activity was consistent with an absence of differences in urine volume and with the apparent utilization of the enhanced ion intake for body functions.

Probable toxicosis in cattle in Israel caused by the oak Quercus calliprinos.

A probable outbreak of oak (Quercus calliprinos) toxicosis in a herd of beef cattle--heifers and first-calving cows--grazing in the Judean foothills of Israel is described. Toxicosis probably occurred because of the consumption of oak leaves and buds during a period of pasture scarcity without any feed supplementation. A progressive syndrome of wasting, dullness, anorexia, polyuria, nephrosis, constipation and recumbency, culminating in death, was seen. A high mortality rate of 83% (38/46 animals) was noted. The clinical-pathological findings revealed increases in blood urea, creatinine, aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), creatine kinase (CK), lactate dehydrogenase (LDH) and inorganic phosphorus. Decreases were found in alkaline phosphatase (ALP), total serum protein, albumin (ALB), triglyceride (TG), calcium (Ca), magnesium (Mg), sodium (Na) and chloride (CI). The main pathological findings were severe nephrosis, chronic interstitial nephritis, and occasional intestinal ulceration. On the basis of epidemiology, clinical signs, clinical-pathological and pathological findings and renal histology, a tentative diagnosis of oak toxicosis was made.

Clinical, clinico-pathological and serological studies of Babesia ovis in experimentally infected sheep.

Clinical, clinico-pathological and serological studies were performed in sheep experimentally infected with Babesia ovis. Acute babesiosis occurred in all the lambs infested with adult Rhipicephalus bursa ticks and in one lamb infested with the larvae. The rate of parasitaemia and the degree of anaemia were not correlated. Decrease in the packed-cell volume ranged from 30 to 40%. Parasitized erythrocytes were not observed to block capillaries in the brain, which explained the absence of nervous symptoms in acute babesiosis. The kidneys were the most severely affected organs, exhibiting acute glomerulonephritis. The lesions observed were suggestive of vascular alteration and vascular stasis, leading to anoxia of the tissues. A disseminated intravascular coagulation (DIC) syndrome was recorded in sheep infected with babesiosis. A marked increase in the enzymes of the transaminase groups, mainly aspartate aminotransferase (AST), was observed. Enzymatic changes (increases in AST, alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) and decreases in sorbitol dehydrogenase (SDH), alkaline phosphatase (ALP) and malic enzyme (MEZ)), decreases in total proteins and albumin, and increases in urea and creatinine might reflect the degree of severity of the damage to the liver and kidney tissues. Most of the lambs (85%) that were infested with larvae, and all lambs infested with adult R. bursa ticks, reacted serologically to B. ovis antigen. The serological reactions following infestation with the larvae occurred much later than those following infestation with the adult stage. The lambs which were infested with larvae showed mild clinical reactions when challenged by infected R. bursa adults, as compared with the reactions to the challenge in naive control animals. The serological findings, in addition to the fact that one splenectomized lamb reacted to larval infestation with acute ovine babesiosis, show that the preimaginal stages of R. bursa can transmit B. ovis, usually causing a sub-clinical disease. It is suggested that infections derived from preimaginal ticks in the winter can preimmunize sheep for the subsequent more severe infections derived from adult ticks in the summer. Furthermore, in the absence of a reliable vaccine against B. ovis, grazing flocks in the enzootic regions should be exposed to the preimaginal stages during their activity period (October-February) before exposure to the adult ticks in spring and summer (April-July).

Gamma-glutamyltransferase as a marker for the pasteurization of raw milk.

The degree and rate of inactivation of gamma-glutamyltransferase in raw cow's milk by heating at 50, 60, 70, and 80 degrees C for 1, 2, 3, 5, 10, 15, 20, 25, and 30 min were measured to evaluate the suitability of this enzyme as a marker for the pasteurization of milk. The enzymes alkaline phosphatase and lactate dehydrogenase were also measured under similar conditions for comparison. The patterns of heat inactivation of gamma-glutamyltransferase and alkaline phosphatase were similar, with only a minimal inactivation of the enzymes at 50 degrees C. The rate of inactivation increased as a result of increasing temperatures and time. A complete inactivation of both enzymes was seen at 70 degrees C after 10 min and at 80 degrees C after 1 min. Lactate dehydrogenase showed a higher heat resistance with almost complete inactivation at 70 degrees C for 30 min, and compete inactivation at 80 degrees C for 3 min. No activities of these enzymes were found in commercially pasteurized or heat-treated milk. The levels of gamma-glutamyltransferase in raw milk were between 8 and 10% higher than those of alkaline phosphatase and lactate dehydrogenase, making it more sensitive and accurate as a testing marker. It seems that gamma-glutamyltransferase may serve as a good pasteurization marker. Furthermore, the simplicity of testing and the availability of commercial kits for testing by both wet and dry chemistry make it an attractive choice, especially because dry chemistry procedures overcome the difficulties originating from the turbidity of milk, which interferes with spectrophotometric procedures.

The toxic effects of monensin and chloramphenicol on laying turkey breeder hens.

A case report of choramphenicol and monensin poisoning in turkey breeder hens is presented in which anorexia and a fall in egg production were features. An experiment study was then conducted in turkey breeding hens given increasing levels of monensin and chloramphenicol singly or in combination. Monensin fed at levels of 42 or 85 ppm had no adverse effect on egg production but chloramphenicol in the drinking water at 500 mg/l for 4 days caused a 9.2% fall in egg production compared to control untreated birds. When 500 mg/l chloramphenicol and 42 ppm monensin were given together for 8 days there was lameness, 12% mortality, but no drop in egg production. When 70 ppm monensin and 500 mg/l chloramphenicol were given together for 4 days, 14 of 22 birds died and egg production in the remainder ceased. Withdrawal of monensin arrested mortality but egg production did not recover. Serum creatine phosphokinase levels in this group were 20 times greater than those of the controls and were similar to birds studied in the case report.

Acute maduramicin toxicity in calves.

A herd of 277 beef-breed calves in three age groups was mistakenly given the poultry coccidiostat maduramicin in a total mixed ration. It caused an acute toxicosis in which sudden death was the sole clinical finding in most cases. One group of 212 calves aged five to eight months suffered a mortality of 51 per cent in eight days and a total mortality of 56 per cent during the 40 days in which mortality was recorded. Mortality of only 3 per cent was recorded in two other groups of calves aged nine to 16 months in eight days and a total mortality of 11 per cent over the 40-day period.

Characterization of the subclinical phase of canine ehrlichiosis in experimentally infected beagle dogs.

Beagle dogs were examined during the subclinical phase of canine ehrlichiosis under controlled conditions. Emphasis was placed on gathering data before artificial inoculation with Ehrlichia canis, and comparing these data with those of the subclinical phase of the disease. In this study all dogs were clinically healthy throughout the 6 month examination period. All subclinically infected dogs had IFA antibody titers to E. canis at a dilution varying from 1:2560 to 1:20480. The most prominent haematological finding was mild thrombocytopenia with a concomitant increase in platelet size, seen in eight of the nine dogs examined. Leukocyte counts were statistically significantly reduced in 78% of the dogs, compared with their preinfection values, with 71% of dogs having significantly reduced absolute neutrophil counts. None of the dogs were either leukopenic nor neutropenic. Six of the nine dogs had increased serum gamma-globulin concentrations. No dogs were overtly anemic, although declines in packed cell volume, haemoglobin concentration and total erythrocyte count were detected in an inconsistent manner among the dogs. It was concluded that, the most reliable parameters for judging possible subclinical ehrlichial infection in beagle dogs was mild thrombocytopenia, together with a persistently high antibody titer to E. canis. Hypergammaglobulinemia would increase the suspicion further. Based on the results presented, routine testing of dogs in E. canis endemic areas is recommended in order to identify and treat dogs in the subclinical phase of the disease.

Severe hepatopathy in geese and broilers associated with ochratoxin in their feed.

The feeding of a shipment of imported corn was associated with a severe reduction in growth and increased mortality in geese, and increased mortality in broilers. Pathological examinations revealed hepatopathy, visceral gout and mild nephropathy in geese, and in broilers an hepatopathy, which was often severe, and ascites. Samples of feed from affected geese farms were examined for up to 24 mycotoxins, and ochratoxin was found in 6 of 15 samples at levels up to 930 ng/g. The syndrome was experimentally reproduced by feeding geese and broilers suspect feeds with the natural ochratoxin contamination. It is believed that another, unidentified, mycotoxin was the major cause of the hepatotoxicity, and that ochratoxin served in this case as an indicator of a multi-mycotoxin involvement.

Sex and genotype dependence on the effects of long-term high environmental temperatures on cellular enzyme activities from chicken organs.

The effects of long term hyperthermia on enzyme levels in the chicken heart and breast muscles, brain, kidney, liver and lung, in relation to sex and degree of feathering, were studied. The enzymes studied were alanine and aspartate amino-transferases, alkaline phosphatase, creatine kinase, lactic dehydrogenase and gamma-glutamyltransferase. Double heterozygote frizzled naked neck and normally feathered male and female broilers were exposed to 24 degrees C (control group) and 32 degrees C (experimental group), for 5 weeks, starting at the age of 3 weeks. The birds were killed, the tested organ removed, homogenized and cell-free supernatant was obtained by centrifugation. Enzyme activities were measured with an autoanalyser and specific activities were calculated. Prolonged heat stress resulted in changes of enzyme activities in all the tissues studied. No significant differences were seen in the cellular enzyme levels from the various organs between male and female birds unexposed to heat stress. Following heat stress, however, greater changes in enzyme levels were seen in the brain, heart muscle and kidney of males compared to females. No significant differences were seen in the cellular enzymes studied in the tissues between the normal and frizzled naked neck chicken. Following prolonged heat stress, there were some differences in the degree of response between the frizzled naked neck and control groups. These differences did not show a consistent or clear pattern indicative of the degree of stress in each of the groups.

Age-dependent variations of lactate dehydrogenase and creatine kinase activities in water buffalo calf serum.

The electrophoretic patterns of the serum enzymes lactate dehydrogenase and creatine kinase from water buffalo calves are described. Differences in total activities as well as their relative distribution were seen at ages ranging from 1 to 10 weeks. While total lactate dehydrogenase activity increased by over 100%, total creatine kinase increased by almost 400%. The relative activities of lactate dehydrogenase 1 and 5 decreased with age. Lactate dehydrogenase 2 and 3 increased and lactate dehydrogenase 4 did not change. In relation to creatine kinase, the prevalent isoenzyme was creatine kinase-MM, but it's relative activity gradually decreased in comparison to the other two isoenzymes (creatine kinase-MB and creatine kinase-BB). Creatine kinase-BB was completely absent until the 3rd week of age. The percentage modifications of creatine kinase isoenzymes were correlated to age. The results suggest that isoenzymatic separation and characterization of lactate dehydrogenase and creatine kinase in relation to the various tissues can significantly contribute to the diagnosis of diseases which are linked to tissue damage.

gamma-Glutamyltransferase and serum proteins in buffalo calves following colostral ingestion.

Catalytic activity concentrations of gamma-glutamyltransferase in blood were determined for the evaluation of colostral ingestion. gamma-Glutamyltransferase levels were compared with those of total proteins, albumins and total globulins in the serum, showing good correlation at 2-3 days after birth. gamma-Glutamyltransferase was determined using the Boehringer Mannheim "Reflotron" dry chemistry enzyme strips. After colostral ingestion, blood gamma-glutamyltransferase increased from 35 +/- 19 U/l at birth to 1563 +/- 1283 U/l. Electrophoretic separation of total proteins showed an increase of serum gamma-globulins from 4.2 g/l before ingestion to 32.9 g/l after ingestion. The same analytes were determined in colostrum showing the major protein fraction to be gamma-globulins. gamma-Globulins and gamma-glutamyltransferase permeate to the blood stream, and there is a good correlation between them. The enzyme gamma-glutamyltransferase can be used as an equivalent for the determination of gamma-globulins in the calf serum and the degree of colostral ingestion.

Serum protein alterations in canine ehrlichiosis.

Serum protein electrophoresis was performed in 42 dogs with naturally occurring Ehrlichia canis infection and in 15 clinically healthy dogs (control dogs). The infected dogs were found to have a significant hypoalbuminaemia, hyperglobulinaemia and hypergammaglobulinaemia compared to the control dogs (P < 0.001). A polyclonal gammopathy was found in all but one of the infected dogs which presented a monoclonal gammopathy. alpha-1 globulin was lower while alpha-2 and beta-2 globulin concentrations were significantly higher in the infected dogs (P < 0.0001, P < 0.05 and P < 0.005, respectively). The infected dogs were divided into two subgroups according to haematological parameters, defined as pancytopenic (n = 13) and non-pancytopenic (n = 29). When compared, the pancytopenic group revealed significantly lower concentrations of total protein, total globulin and gammaglobulin (P < 0.01, P < 0.05 and P < 0.005 respectively). The lower concentrations of the gammaglobulins coupled with the pancytopenia suggest that the immune state of the pancytopenic E. canis infected dogs is more compromised, and therefore secondary infections should be expected more frequently in these dogs.