Changes in the concentrations of bile acids in the plasma of sheep with liver damage

The concentration of total plasma bile acids was measured in normal sheep and in sheep in which liver damage was induced by chronic copper poisoning, ligated bile ducts or induced ketosis. All three treatments produced a rise in total bile acid concentration in plasma which was proportional to the degree of hepatic damage seen histologically and which tended to parallel changes in activity of iditol, and glutamate dehydrogenase and aspartate amino-transferase in plasma. Plasma bile acid concentration was a more sensitive method of detecting these types of liver damage than was the measurement of total plasma bilirubin concentration, and could be used to assess alterations in liver function in sheep.     

Copper poisoning in four llamas

Copper poisoning developed in 1 adult and 3 juvenile llamas after excessive dietary intake of copper resulted in an incorrect copper:molybdenum ratio. Total dietary copper was determined to be 36 mg/kg of feed, with a copper:molybdenum ratio of 16.6:1. Clinical signs associated with the toxicosis included acute onset of anorexia and lethargy. Liver enzyme activities (aspartate amino transaminase, lactate dehydrogenase, gamma-glutamyl transferase) and serum copper concentration were high in specimens obtained within 48 hours before death. Gross necropsy findings were limited to mild hepatomegaly. Histologically, hepatic lesions included acute massive necrosis of hepatocytes with and without bile duct proliferation, double hepatic plates with loss of orientation, anisocytosis, anisokaryosis, and an intralobular mosaic pattern of necrosis involving half of the hepatocytes. Analysis of hepatic copper concentrations suggested that juvenile llamas develop signs of poisoning at lower hepatic copper concentrations, compared with adults.     

Evaluation of total plasma bile acid concentrations for the diagnosis of hepatobiliary disease in horses

Plasma bile acid concentrations were measured in normal horses. There was no diurnal variation in values, and age and sex had no effect. There was no significant difference between serum and plasma bile acid concentrations in clinically normal horses. Plasma bile acids were stable on storage for one month at -20 degrees C. The total plasma bile acid concentrations together with total and direct bilirubin concentrations and plasma activities of aspartate aminotransferase, glutamate and iditol dehydrogenase were evaluated in horses with various types of hepatobiliary disease (hepatic necrosis, lipidosis, neoplasia and cirrhosis), gastrointestinal disease, cardiovascular, orthopaedic and various other conditions not affecting the liver. Total plasma bile acids together with plasma glutamate and iditol dehydrogenase activities were the best indicators of liver disease. Total plasma bile acid concentrations were the most sensitive indicator of a wide variety of hepatic diseases but alone were unhelpful in differential diagnosis and were of more value when combined with the other tests of hepatic disease.     

A screening test for subclinical liver disease in horses affected by pyrrolizidine alkaloid toxicosis

Objective: To evaluate various biochemical tests as indicators of subclinical liver disease in horses exposed to pyrrolizidine alkaloid toxicosis.
Design: A clinical pathology field study.
Animals: Twenty-two clinically normal horses from four properties in the Kimberley region of Western Australia.
Procedure: Serum samples from each horse were assayed for gamma glutamyltransferase, alkaline phosphatase and aspartate aminotransferase activities, and for serum bile acid concentration, albumin and total protein. Serum protein electrophoresis was performed and their amino acid profiles determined. Bromosulphophihalein halfclearance times were measured. Horses were then subjected to a single liver biopsy. Results were analysed by, variance of group means, the Fisher-Irwin exact test, and by sensitivity and specificity calculation.
Results: Horses were classified into 2 groups, of 10 unaffected and 12 subclinically affected, on the basis of liver histology. Significant differences between the unaffected and subclinical groups were observed for gamma glutamyltransferase and alkaline phosphatase activities (P < 0.01). Gamma glutamyltransferase had sufficient sensitivity (75%) and specificity (90%) to function as a primary screening test for subclinical liver disease in horses exposed to pyrrolizidine alkaloids. Alkaline phosphatase was useful, but with lower sensitivity (58%).
Conclusion: Serum gamma glutamyltransferase activity is a useful screening test for detecting subclinical liver disease in horses exposed to pyrrolizidine alkaloids under field conditions in northern Australia.     

The excretion of phylloerythrin and bilirubin by calves and sheep.

Under general anaesthesia the common bile duct was ligated in two sheep and two calves. Occlusion of the duct was permanent and was followed by portal fibrosis, proliferation of bile ducts and intrahepatic bile stasis. Mild hepatic cell damage was accompanied by the release of glutamate dehydrogenase, sorbitol dehydrogenase and arginase into serum. The release of gamma-glutamyl transpeptidase was slower but more continuous. One sheep and one calf developed peritonitis associated with the leakage of bile from a biopsy wound in the live. One of these animals and the other two on which biopsy was not performed became photosensitised on exposure to sunlight. The concentration of phylloerythrin was high in serum and urine. All animals became jaundiced and the increased concentration of bilirubin in serum and urine was mainly direct reacting, ie, conjugated with glucuronic acid.     

Serum bile acids as an indicator of liver disease in dogs

Total serum bile acids were determined in 62 dogs with different primary or secondary liver diseases, using 3α-hydroxysteroid dehydrogenase coupled to nitrobluetetrazolium in a centrifugal analyzer. A reaction time of 4 min was sufficient, yielding a within run coefficient of variation of 7% at 6 µmol/1 and 3% at 27 µmol/1. A reference range of 0–4.4 µmol/1 2 h post prandially was observed. The sensitivity of bile acids as a liver function test was superior to that of alanine and aspartate aminotransferase, alkaline phosphatase, γ-glutamyltransferase and combinations of two of these. The bile acids test detected 36 of 39 patients with a morphological or clinical liver diagnosis. For dogs with heart failure the bile acids test was a markedly more sensitive indicator of secondary liver involvement than alanine aminotransferase or alkaline phosphatase. For secondary liver affections associated with pyometra or epilepsy medication the opposite was the case. Bile acid values in the pooled patient material was not correlated to any of the 4 enzymes measured. For cirrhosis there was positive correlation, however, with the amino transferase values.     

Hepatic Lipidosis in Anorectic, Lactating Holstein Cattle: A Retrospective Study of Serum Biochemical Abnormalities

The association between hepatic lipidosis (HL) and disease in 59 anorectic, ketotic, lactating Holstein heifers and cows was investigated. Severe HL, as determined by histologic evaluation of liver tissue, was present in 46 animals; only half of these animals required intensive treatment for ketosis, and only half had serum biochemical evidence of liver disease, as determined by the presence of a test value of 2-fold or greater than the upper limit of the reference range for at least 2 of the 4 serum tests: gamma-glutamyl transferase, aspartate aminotransferase, and sorbitol dehydrogenase activities and bile acid concentrations. Most cattle with biochemical evidence of liver disease and severe HL had been lactating for 14 or more days. Cows that required intensive treatment inconsistently had serum biochemical evidence of liver disease.
Although cattle with severe HL had significantly higher serum bilirubin concentrations and aspartate aminotransferase and sorbitol dehydrogenase activities than cattle with less severe lipidosis, the specificity of abnormally high serum sorbitol dehydrogenase activity or bilirubin concentration for severe lipidosis was only 8%. Abnormally high serum aspartate aminotransferase activity was 83% sensitive and 62% specific for severe lipidosis. Serum glucose and total carbon dioxide concentrations were significantly lower in cattle with severe lipidosis than in those with mild or moderate lipidosis, and low serum glucose or total carbon dioxide concentrations were rare in cattle without severe lipidosis. From these data, we conclude that the use of a single biochemical or histopathologic criterion to define severity of disease or degree of liver compromise in anorectic, ketotic cows results in the misidentification of many animals.     

Clinicopathologic study of horses surviving pyrrolizidine alkaloid (Senecio vulgaris) toxicosis

Twenty horses of various ages had inadvertently ingested alfalfa hay contaminated with Senecio vulgaris. Among them, 4 died of liver disease. Blood was collected from affected horses at monthly intervals for 7 months and at the 9th and 14th months. The following serum enzymes and chemical items were assayed: aspartate aminotransferase, lactate dehydrogenase, alkaline phosphatase, gamma-glutamyl transferase, sorbitol dehydrogenase, total bilirubin, BUN, glucose, cholesterol, inorganic phosphate, calcium, total protein, and albumin. Amino acid profiles, conjugated bile acids, sulfobromophthalein clearance times, and liver histopathologic changes via serial biopsies were also monitored. Liver histopathologic changes revealed lesions progressively increasing in severity. Aspartate aminotransferase and plasma amino acid ratios indicated chronic liver degeneration (0.05 level of significance). gamma-Glutamyl transferase and lactate dehydrogenase as well as BUN values fluctuated, but returned to within reference values. Horses appeared clinically normal 14 months after intoxication, but were unable to tolerate stress of exercise.     

Serum and tissue enzyme profiles of goats

Serum enzyme activities of sorbitol dehydrogenase, glutamate dehydrogenase, gamma glutamyltransferase, alkaline phosphatase, aspartic aminotransferase, and creatine kinase, were measured in five clinically normal mixed-breed goats. Tissue activities of these enzymes were also measured in two goats. These basal serum values were then used to determine the response to treatment with carbon tetrachloride (CCl4). The basal value for serum and hepatic tissue sorbitol dehydrogenase were appreciably greater for goats than previously reported for sheep and cattle. The change in the above serum enzymes after CCl4 treatment resembled that in sheep, but the amount of sorbitol dehydrogenase increase was less than that in sheep. This study established basal tissue and serum enzyme activity values and demonstrated the efficacy of the use of changes in serum S.D.H. and G.D.H. activity as indicators of acute hepatopathy in goats.     

Evaluation of ornithine carbamoyl transferase and other serum and liver-derived analytes in diagnosis of fatty liver and postsurgical outcome of left-displaced abomasum in dairy cows

OBJECTIVE: To evaluate postsurgical outcome in dairy cows with left-displaced abomasum (LDA) with regard to severity of fatty liver and assess the usefulness of preoperative determination of serum ornithine carbamoyl transferase (OCT) activity, bile acids concentration, and other variables for evaluating liver function during the postsurgical convalescence period. DESIGN: Prospective study. ANIMALS: 68 Holstein cows. PROCEDURES: Blood and liver biopsy specimens were obtained during standing LDA surgery. Liver tissue was examined histologically and classified by severity of fatty change. Serum activities of liver-derived enzymes and concentrations of total lipids, triglycerides, bile acids, glucose, beta-hydroxybutyric acid, bilirubin, and nonesterified fatty acids were determined. RESULTS: Most cows with LDA and cows with severe fatty liver were detected within the first month after calving. Postsurgical outcome was related to severity of fatty liver. All cows that died had severe fatty liver. Serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration were sensitive indicators of fatty liver. Serum bile acids concentration was not an accurate indicator of fatty liver. CONCLUSIONS AND CLINICAL RELEVANCE: Postsurgical outcome of cows undergoing surgery to correct LDA was related to fatty liver severity. Assessment of serum activities of OCT, aspartate aminotransferase, and glutamate dehydrogenase and serum total bilirubin concentration is recommended for diagnosis of fatty liver in dairy cows with LDA, whereas determination of bile acids concentration is not. The strong correlation between OCT activity and degree of hepatocellular damage supports use of this enzyme for assessing severity of fatty liver and predicting postsurgical outcome in cows with LDA.     

Protective effects of methanolic leaf extracts of Monanthotaxis caffra against aflatoxin B1-induced hepatotoxicity in rats

Aflatoxins are potent hepatotoxic and carcinogenic secondary metabolites produced by toxigenic fungi. The present study investigated the protective effect of methanolic leaf extracts of Monanthotaxis caffra (MLEMC) against aflatoxin B1-induced toxicity in male Sprague-Dawley rats. The rats were randomly divided into 6 groups of 8 animals each. Five groups were administered orally for seven days with three different concentrations of MLEMC (100 mg/kg, 200 mg/kg and 300 mg/kg), curcumin (10 mg/kg) or vehicle (25% propylene glycol). The following day, these groups were administered 1 mg/kg b.w. of aflatoxin B₁ (AFB₁). The experiment was terminated three days after administration of AFB₁. Group 6 represented untreated healthy control. Serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, creatinine and liver histopathology were evaluated. Methanolic leaf extracts of M. caffra decreased the levels of aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and creatinine in the sera of rats as compared with the AFB₁ intoxicated group. Co-administration of MLEMC improved the histological characteristics of the hepatocytes in contrast to the AFB₁ treated group, which had mild to severe hepatocellular injuries including bile duct proliferation, bile duct hyperplasia, lymphoplasmacytic infiltrate and fibrosis. Extracts of M. caffra were beneficial in mitigating the hepatotoxic effects of AFB₁ in rats by reducing the levels of liver enzymes and preventing hepatic injury.     

Acute aflatoxicosis in swine: clinical pathology, histopathology, and electron microscopy

Feeder pigs weighing 12 to 15 kg each were given a single oral dose of aflatoxin, 1.2 mg/kg of body weight. Liver-specific serum enzyme activities were compared with gross, microscopic, and ultrastructural hepatic changes in individual pigs euthanatized at 24, 48, and 72 hours after they were given aflatoxin. The greater the morphologic change in liver of the treated pigs, the greater the increase in liver-specific serum enzyme activities. Isocitric dehydrogenase, alkaline phosphatase, sorbitol dehydrogenase, and aspartate aminotransferase activities increased in 6 of 8 treated pigs by 24 hours. Increase in gamma-glutamyl transpeptidase activity was not significant. Microscopic and ultrastructural changes in centrilobular hepatocytes included glycogen deletion, mitochondrial and endoplasmic reticulum swelling, membrane disruption, and nuclear fragmentation at 24 hours. The centrilobular areas had marked extravasation of erythrocytes at 24 hours without basal lamina changes. At 72 hours, the centrilobular hepatocytes had increased lipid vacuoles and acceptable amounts of glycogen. Marked infiltrations of monocytes, plasma cells, and lymphocytes were also present at this time.     

Plasma bile acid elevation following CCI4 induced liver damage in dogs, sheep, calves and ponies.

Plasma bile acid concentration was determined in normal dogs,sheep, calves and ponies for three days before and six days after liver damage, induced by carbon tetrachloride. In all species, a significant increase in plasma bile acid concentration was associated with a concomitant significant increase in plasma sorbitol dehydrogenase and transferase activity. Plasma bilirubin also significantly increased in all animals except the dogs. Results suggested that plasma bile acid levels could be used to test liver function in domestic animals.     

Copper toxicosis in a Boer goat

A 1‐year‐old female Boer goat was presented with a 1‐day history of pigmenturia, anorexia, and shivering. Anemia was not present initially, but progressive hemolytic anemia developed subsequently and was characterized by the finding of Heinz bodies in both intact RBCs and in ghost cells and the presence of atypical fusiform RBCs. Plasma biochemical analysis revealed increased activities of aspartate aminotransferase and gamma‐glutamyltransferase, hyperbilirubinemia, and azotemia. Histopathologic examination of a liver biopsy revealed necrosis of individual hepatocytes and intracytoplasmic rhodamine‐positive granules, consistent with copper. Copper concentration in ante‐mortem hepatic tissue was increased, and a diagnosis of copper toxicosis was made. Despite supportive therapy, the goat continued to decline and was euthanized. Necropsy findings included hepatic necrosis and hemoglobinuric nephrosis. Freshly collected specimens of liver and kidney had markedly increased copper concentrations. The mineral composition of the water, grass hay, and goat chow was evaluated, and toxins and significant mineral imbalances were not found. The underlying cause of the hepatic accumulation and subsequent release of copper remains unclear in this goat. Recently, Boer goats have been recognized as being prone to copper toxicosis and may be more susceptible than other breeds; similar to sheep, Boer goats may experience a hemolytic crisis secondary to copper toxicosis.     

Plasma bile acids, lactate dehydrogenase and sulphobromophthalein retention test in canine carbon tetrachloride intoxication

This paper studies the progression of various hepatic function tests (fasting and post prandial plasma bile acids, lactate dehydrogenase [LDH], hydroxybutyrate dehydrogenase [HBDH], LDH isoenzymogram, and 45 minutes sulphobromophthalein retention) in a classic model of canine hepatic injury. Sulphobromphthalein and plasma bile acid determination are excellent indicators of hepatic damage. Post prandial bile acid quantification does not improve the diagnostic capacity of this test. LDH alters only in the first 48 hours after poisoning and the study of the HBDH/LDH ratio makes it possible to differentiate the origin of plasma LDH. LDH isoenzymogram is not a practical test for routine clinical diagnosis.     

Clinicopathologic evaluation of hepatic lipidosis in periparturient dairy cattle

BACKGROUND: Fatty change of the liver (FCL) is very common in dairy cattle periparturiently. Many laboratory methods have been implicated in order to assist the diagnosis. HYPOTHESIS: To investigate whether FCL in dairy cattle could be evaluated by assessment of ornithine carbamoyl transferase (OCT) by means of an assay modified for bovine serum, other enzyme activity, serum bile acids (SBA) concentration, or other biochemical constituents. ANIMALS: A total of 187 dairy cattle were included: 106 were suspected to have liver dysfunction and were examined after referral by veterinarians; 70 were clinically healthy with mild FCL; and 11 were clinically healthy without FCL. METHODS: Blood and liver biopsy samples were obtained after clinical examination. Histologic examination by light microscopy and classification of samples according to the severity of FCL was done, and total lipid and triglyceride concentration was measured. In serum, OCT, aspartate aminotransferase (AST), alanine aminotransferase (ALT), sorbitol dehydrogenase (SDH), glutamate dehydrogenase (GDH), alkaline phosphatase (ALP), and gamma-glutamyltransferase (gamma-GT) activity as well as SBA, glucose, ketones, total bilirubin (tBIL), and nonesterified fatty acids (NEFA) concentration were measured. RESULTS: OCT and AST activity and tBIL concentration correlate well with the degree of FCL. SBA concentration does not contribute well to FCL diagnosis. The majority of FCL cases appeared within the first 21 days-in-milk (DIM). The majority of moderate-to-severe and severe FCL cases arose in the first 7 DIM. CONCLUSIONS AND CLINICAL IMPORTANCE: Except for OCT, AST, and tBIL, none of the biochemical tests used, including SBA, had sufficient discriminatory power to differentiate reliably between mild and severe FCL because of poor sensitivity. A weak correlation between clinical signs and the extent of FCL was evident.     

Plasma amino acid analysis in dogs with experimentally induced hepatocellular and obstructive jaundice

Plasma amino acid concentrations were determined weekly for 4 weeks in 6 sham-operated control dogs, in 6 sham-operated dogs after induction of liver disease with dimethylnitrosamine (DMNA), and in 6 dogs after surgical ligation of the common bile duct. Results were compared with standard biochemical indices of liver function and with histologic changes in serial liver biopsy specimens. Concentrations of most amino acids increased after 2 weeks in DMNA-treated dogs, whereas they were normal or decreased in bile duct-ligated dogs. With increasing severity of the liver disease, the molar ratio (normal mean +/- SEM = 3.48 +/- 0.14) in DMNA-treated dogs decreased to 2.42 +/- 0.19 by 2 weeks and to 1.17 +/- 0.09 by 4 weeks. The ratio remained normal in bile duct-ligated dogs. Molar ratio and aromatic amino acid concentrations correlated better with hepatic necrosis and inflammation than did standard biochemical indices (eg, bilirubin, liver enzymes). Therefore, plasma amino acid analysis and determination of the molar ratio may be useful in the differential diagnosis of hepatocellular and obstructive jaundice in dogs. A decrease in the molar ratio may reflect portal hypertension and hepatocellular disease.     

Increased Serum Alanine Aminotransferase Activity Associated With Muscle Necrosis in the Dog

Serum activity of alanine aminotransferase (ALT) was consistently increased in dogs with canine X-linked muscular dystrophy (CXMD), a primary myopathy characterized by profound and on-going skeletal muscle necrosis. In order to determine whether the ALT was of liver origin, serum activity of creatine kinase (CK), aspartate aminotransferase (AST), ALT, and sorbitol dehydrogenase (SDH) obtained from dystrophic dogs was compared with enzyme activity present in clinically normal dogs. In dystrophic dogs at all ages tested, serum activity of CK, AST, and ALT was increased, and significant increases were present in dogs four weeks or older. In contrast, SDH activity in dystrophic dogs was not statistically different from values in clinically normal dogs. Ultrastructural examination of liver tissue revealed no evidence of hepatic degeneration in dystrophic dogs. It was concluded that increased serum activity of ALT in the dog may be associated with severe skeletal muscle degeneration, without concurrent hepatocellular necrosis.     

Metabolic changes and induction of hepatic lipidosis during feed restriction in llamas.

OBJECTIVES: To determine whether feed restriction induces hepatic lipidosis (HL) in llamas and to evaluate the metabolic changes that develop during feed restriction. ANIMALS: 8 healthy adult female llamas. PROCEDURE: Llamas were fed grass hay at a rate of 0.25% of their body weight per day for 13 to 28 days. Llamas were monitored by use of clinical observation, serum biochemical analyses, and ultrasound-guided liver biopsies. RESULTS: All 8 llamas lost weight and mobilized fat. Five llamas developed HL, including 4 that were nursing crias. During the period of feed restriction, mean serum concentration of bile acids and activities of aspartate aminotransferase (AST), sorbitol dehydrogenase (SDH), and gamma-glutamyl transferase (GGT) were significantly higher in llamas that developed HL, compared with llamas that did not. Mean insulin-to-cortisol concentration ratios were lower in llamas with HL before and up to 7 days of feed restriction, compared with those that did not develop HL. CONCLUSIONS AND CLINICAL RELEVANCE: HL in llamas may be induced by severe feed restriction, particularly in the face of increased energy demand. Llamas with weight loss attributable to inadequate dietary intake may develop biochemical evidence of hepatopathy and HL. Increases in serum concentration of bile acids and activities of GGT, AST, and SDH may indicate the development of HL in llamas and identify affected animals for aggressive therapeutic intervention.