Comparison of biochemical values in serum and plasma, fresh and frozen plasma, and hemolyzed samples from orange-winged Amazon parrots (Amazona amazonica)

BACKGROUND: To the authors' knowledge, on the basis of sample type, storage condition, or hemolysis, differences in serum and plasma biochemical values have not been evaluated in orange-winged Amazon parrots (Amazona amazonica). OBJECTIVES: The purpose of this study was to compare values for biochemical analytes in serum vs plasma, fresh vs frozen plasma, and nonhemolyzed vs hemolyzed samples in orange-winged Amazon parrots. We also compared differences in serum and plasma yield from whole-blood aliquots. METHODS: Fifteen biochemical analytes were evaluated in paired serum and plasma, fresh and frozen plasma, nonhemolyzed and hemolyzed serum and plasma samples from orange-winged Amazon parrots (n = 10) using a wet reagent analyzer. Hemolysis was assessed qualitatively (visually) and quantitatively (hemoglobin [Hgb] measured spectrophotometrically). Serum and plasma yields from 500-microl whole-blood aliquots were determined from centrifuged samples. RESULTS: Analyte values significantly differed among sample groups, but were still within published reference intervals, with the exception of increases in potassium concentration in markedly hemolyzed serum and plasma samples. Clinically important changes in hemolyzed serum and plasma samples included increases in potassium, phosphorus, and albumin concentrations and lactate dehydrogenase activity. The degree of hemolysis assigned qualitatively did not correlate with quantitative Hgb concentration. A significantly greater yield of plasma (288 +/- 13 microL) than serum (241 +/- 44 microL) was obtained. CONCLUSIONS: Significant differences may occur in different sample types, however, only changes in potassium, phosphorus, albumin, and lactate dehydrogenase values in hemolyzed samples were considered clinically relevant. Lack of agreement between qualitative and quantitative Hgb concentration indicates the unreliability of visual estimation. Based on higher sample yield, and lack of clinically relevant differences from serum, plasma is a better sample choice for clinical chemistry analysis in birds.     

Hemolysis as a factor in clinical chemistry and hematology of the dog

Serum biochemical, hemostatic, and hematologic analytes were determined by four laboratories on dog serum or plasmas containing increasing amounts of hemolysate. From the results of testing, interferographs were prepared to aid in decision making and to enhance visualization of the effects of hemolysis on the determination of the analytes. Heniktsus consistently interfered with the analysis of creatinine phosphokinase, lactic dehydrogenase, aspartate aminotransferase, lipase, and albumin, all of which appeared to increase with increasing hemolysis. The results for the remainder of the biochemical, hemostatic, and hematologic analytes varied greatly among analyzers or methods, rarely in a predictable manner, indicating that each laboratory should evaluate the effects of hemolysis on each analyzer and for each method used, in order to make informed decisions on the use of hemolyzed but irreplaceable samples.     

Evaluation of five clinical chemistry analyzers for use in health assessment in sea turtles

OBJECTIVE: To compare blood biochemical values obtained from a handheld analyzer, 2 tabletop analyzers, and 2 diagnostic laboratories by use of replicate samples of sea turtle blood. DESIGN: Validation study. ANIMALS: 22 captive juvenile sea turtles. PROCEDURES: Sea turtles (18 loggerhead turtles [Caretta caretta], 3 green turtles [Chelonia mydas], and 1 Kemp's ridley turtle [Lepidochelys kempii]) were manually restrained, and a single blood sample was obtained from each turtle and divided for analysis by use of the 5 analyzers. Hematocrit and concentrations or activities of aspartate aminotransferase, creatine kinase, glucose, total protein, albumin, BUN, uric acid, P, Ca, K, Na, Cl, lactate dehydrogenase, and alkaline phosphatase were determined. Median values for each analyte were compared among the analyzers. RESULTS: Significant differences were found among the analyzers for most values; however, data obtained from the 2 diagnostic laboratories were similar for all analytes. The magnitude of difference between the diagnostic laboratories and in-house units was > or = 10% for 10 of the 15 analytes. CONCLUSIONS AND CLINICAL RELEVANCE: Variance in the results could be attributed in part to differences in analyzer methodology. It is important to identify the specific methodology used when reporting and interpreting biochemical data. Depending on the variable and specific case, this magnitude of difference could conceivably influence patient management.     

Effects of heparin, citrate, and EDTA on plasma biochemistry of sheep: Comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals, but limited information is existing for sheep plasma biochemistry. Ten clinically healthy Baloochi breed of sheep were blood sampled in different tubes containing each anticoagulants and plain tube for harvesting plasma and serum. The concentrations of glucose, cholesterol, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, and magnesium and the activity of aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of GGT, bilirubin and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than that of serum. For corrected citrated plasma significant differences were seen for the concentrations of glucose, creatinine, calcium and the activity of ALP.Most parameters did not show any difference, but significant increase was seen for albumin concentration when heparin was used as an anticoagulant. Using EDTA as anticoagulant caused a significant difference for the concentrations of some of the measured parameters in plasma except glucose, GGT, cholesterol, albumin, bilirubin, CK, and inorganic phosphorus comparing with serum.     

Analyte comparisons between 2 clinical chemistry analyzers

The purpose of this study was to assess agreement between a wet reagent and a dry reagent analyzer. Thirteen analytes (albumin, globulin, alkaline phosphatase, alanine aminotransferase, amylase, urea nitrogen, calcium, cholesterol, creatinine, glucose, potassium, total bilirubin, and total protein) for both canine and feline serum were evaluated. Concordance correlations, linear regression, and plots of difference against mean were used to analyze the data. Concordance correlations were excellent for 8 of 13 analytes (r > or = 0.90); the correlations for albumin, potassium, and calcium were clinically unreliable. The linear regression analysis revealed that several analytes had slopes significantly different from unity, which was likely related to methodological differences. Compared to the wet reagent analyzer, the dry reagent analyzer showed excellent agreement for alkaline phosphatase, alanine aminotransferase, amylase (feline), urea nitrogen, cholesterol, creatinine, glucose, total bilirubin (canine), and total protein. However, it showed only slight to substantial agreement for amylase (canine), calcium, albumin, potassium, and total bilirubin (feline).     

Plasma biochemistry of one-humped camel (Camelus dromedarius): effects of anticoagulants and comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals but limited informations are existing for camel plasma biochemistry. Eleven clinically healthy one-humped camels were blood sampled in different tubes containing different anticoagulants and plain tubes for harvesting plasma and serum. The concentrations of glucose, cholesterol, triglyceride, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, magnesium, and chloride and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of AST, ALT, CK, total bilirubin, and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than in serum. Most parameters did not show any difference, but significant increase for CK activity and significant decrease for GGT, cholesterol, creatinine and chloride were seen when heparin was used as anticoagulant. Using EDTA as an anticoagulant caused a significant difference in the amounts of some measured parameters in plasma except glucose, AST, ALT, GGT, cholesterol, albumin, total protein, bilirubin, and triglyceride in comparison with serum.     

Plasma biochemistry of one-humped camel (Camelus dromedarius): Effects of anticoagulants and comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals but limited informations are existing for camel plasma biochemistry. Eleven clinically healthy one-humped camels were blood sampled in different tubes containing different anticoagulants and plain tubes for harvesting plasma and serum. The concentrations of glucose, cholesterol, triglyceride, total bilirubin, urea, creatinine, total protein, albumin, calcium, inorganic phosphorus, magnesium, and chloride and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK) and gamma glutamyl transferase (GGT) were measured. Except for the amounts of AST, ALT, CK, total bilirubin, and inorganic phosphorus, other measured parameters were significantly lower in citrated plasma than in serum. Most parameters did not show any difference, but significant increase for CK activity and significant decrease for GGT, cholesterol, creatinine and chloride were seen when heparin was used as anticoagulant. Using EDTA as an anticoagulant caused a significant difference in the amounts of some measured parameters in plasma except glucose, AST, ALT, GGT, cholesterol, albumin, total protein, bilirubin, and triglyceride in comparison with serum.     

Effects of Common Anticoagulants on Routine Plasma Biochemistry of Horse and Comparison with Serum

The effects of various types of anticoagulants on plasma biochemistry have been studied in humans and various animals; however, limited information exists for effects on horse plasma biochemistry. Ten clinically healthy Thoroughbred horses were blood sampled in tubes containing different anticoagulants as well as no anticoagulant. The concentrations of glucose, cholesterol, triglyceride, total bilirubin, blood urea nitrogen (BUN), creatinine, total protein, albumin, calcium, inorganic phosphorus, magnesium and iron, and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatine kinase (CK), and gamma glutamyl transferase (GGT) were measured. Except for the amounts of inorganic phosphorus, magnesium, and GGT, other measured parameters were significantly lower in citrated plasma than in serum. When corrected for dilution, significant differences were seen for the amounts of glucose, cholesterol, triglyceride, bilirubin, calcium, iron, and the activity of ALT, ALP, and CK in citrated plasma. Most parameters did not show any difference; however, significant decreases in BUN, total bilirubin, ALT, and CK activity were seen when heparin was used as an anticoagulant. When compared with serum, using ethylenediaminetetra-acetic acid (EDTA) as an anticoagulant produced a significant difference in the amounts of measured plasma parameters with the exception of GGT, albumin, creatinine, inorganic phosphorus, and triglyceride.     

Plasma biochemistry of ostrich (<Emphasis Type="Italic">Struthio camelus</Emphasis>): effects of anticoagulants and comparison with serum

The effects of various types of anticoagulants on plasma biochemistry were studied in man and various animals, but limited information is existing for ostrich plasma biochemistry. Ten clinically healthy ostrich were blood sampled in different tubes containing each anticoagulant and plain tube for harvesting plasma and serum. The concentrations of glucose, cholesterol, uric acid, creatinine, total protein, albumin, calcium, inorganic phosphorus, and magnesium and the activity of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma glutamyl transferase (GGT) were measured. The concentrations of glucose, uric acid, total protein, and calcium were significantly lower in citrated plasma than that of serum. For dilution corrected citrated plasma significant differences were only seen for the concentration of uric acid. Most parameters did not show any differences, but significant increase were seen for glucose, total protein, albumin, and phosphorus concentrations when heparin was used as an anticoagulant.     

Relationship between separation time of plasma from heparinized whole blood on plasma biochemical analytes of loggerhead sea turtles (Caretta caretta)

Concentrations and activities of selected biochemicals of loggerhead sea turtles (Caretta caretta) were determined for plasma that was separated from whole blood samples that were kept up to 96 hr post collection (PC) in a refrigerator. Blood samples collected from seven juvenile captive loggerhead sea turtles were added to tubes containing lithium heparin and were placed on ice. Equal amounts of anticoagulated whole blood from the lithium heparin tubes were then aliquoted into plastic tubes and stored as whole blood under refrigeration until they were centrifuged at 0, 4, 24, 48, and 96 hr PC. Plasma was removed and the analytes that were measured were alkaline phosphatase (ALP), aspartate aminotransferase (AST), gamma glutamyl transferase (GGT), creatine kinase (CK), sodium, chloride, potassium, magnesium, calcium, phosphorus, cholesterol, glucose, urea nitrogen, uric acid, total protein, albumin, and globulin. Compared with values at 0 time, the only analyte to be significantly different at 24 hr PC was GGT (activity decreased by 25%). Compared with values at 0 time, significant differences at 96 hr PC were only seen in AST (2% increase), GGT (25% decrease), glucose (7% decrease), and uric acid (25% increase). Although a statistically significant difference was found in concentrations of phosphorus and cholesterol over time by repeated measures analysis of variance (ANOVA), the follow-up multiple comparison procedure could not define the specific time points at which significant differences occurred. For all other analytes, significant differences over the time course of the study were not found. In these instances, the power of the ANOVA was sufficient (> or = 0.80) to detect any arithmetic differences of a clinically relevant magnitude. Although plasma should be separated from the cellular component of blood as soon as possible PC, in a field situation in which a centrifuge is unavailable, samples can be stored in a portable cooler up to 24 hr without appreciable change in select biochemical analytes.     

Effects of storage time on chemistry results from canine whole blood,heparinized whole blood,serum and heparinized plasma

The stability and storage characteristics of 24 blood constituents from dogs including nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Conditions studied included storing of nonanticoagulated and heparinized whole blood for 3 days (Part A), and storing of serum and heparinized plasma for 3 days (Part B). The storage temperature for both studies was +4 degrees C from day 0 to day 1, and +20 degrees C, from day 1 to day 2 and 3. Eight of 24 analytes showed no significant differences (p > 0.05) for three days in whole blood. However, the stability of all 24 analytes greatly improved by storing serum or heparinized plasma compared to nonanticoagulated or heparinized whole blood. In stored serum or heparinized plasma, 20 of 24 analytes showed no significant differences (p < 0.05) for 3 days. Nine of 24 analytes showed significant differences (p < 0.05) between serum and heparinized plasma, where CK, LDH, GGT, and potassium showed differences of possible clinical importance. This study strongly supports the practice of separating serum/plasma from clot/cells as promptly as possible to achieve improved stability for most analytes under test.     

冷季牧归补饲精料对高海拔地区藏系绵羊血液生化指标的影响

选取9月龄欧拉型藏系绵羊羯羊（30.21 kg±1.42 kg）40只,随机分为4组,A组冷季放牧,B、C和D组冷季牧归每天每只分别补饲0.10、0.20和0.30 kg混合精料,试验期210 d,研究冷季牧归补饲精料对其血清生化指标的影响。结果表明,补饲精料提高了欧拉型藏系绵羊血清总蛋白（TP）、白蛋白（ALB）、球蛋白（GLB）、钙（Ca）的含量,钙/无机磷（Ca/IP）比值及肌酸激酶（CK）、乳酸脱氢酶（LDH）、谷草转氨酶（AST）、谷丙转氨酶（ALT）和γ-谷氨酰胺基转移酶（GGT）活性;随着精料补饲量增大,绵羊血清葡萄糖（GLU）、胆固醇（CHO）和甘油三酯（TG）含量,超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性,生长激素（GH）、胰岛素样生长因子-1（IGF-1）、缩胆囊素（CCK）、胃泌素及免疫球蛋白A（IgA）、免疫球蛋白G（IgG）及免疫球蛋白M（IgM）的浓度增大,但补饲0.30 kg组试验羊IgG浓度较C组略有下降（P>0.05）;与A组相比,补饲精料显著提高了试验羊碱性磷酸酶（AMP）活性、尿素氮（BUN）浓度和BUN/CRE的值,显著降低肌酐（CRE）和尿酸（UA）浓度（P<0.05）。综合上述结果,冷季归牧后补饲适量精料可改善欧拉型藏系绵羊能量和脂肪的代谢能力,增强肝脏蛋白合成,提高机体免疫力和抗氧化力,调节钙磷代谢。     

Effects of in vitro hemolysis on serum biochemistry values of the bottlenose dolphin (Tursiops truncatus).

The effects of in vitro hemolysis on 23 biochemical analytes were assessed in sera from 14 clinically healthy Atlantic bottlenose dolphins (Tursiops truncatus). Each serum sample was divided into three portions for analysis: 1) nonhemolyzed control; 2) moderate hemolysis, simulated by adding hemolyzed serum to a final concentration of approximately 150 mg/dl Hb; and 3) severe hemolysis, simulated by adding hemolyzed serum to a final concentration of approximately 500 mg/dl Hb. Moderate hemolysis resulted in statistically significant increases in the mean values of iron, lactate dehydrogenase, potassium, and uric acid and a decrease in creatinine (P < 0.001). Severe hemolysis resulted in statistically significant changes in the mean values of the above analytes in addition to the following increases: alanine aminotransferase, calcium, and serum globulins (P < 0.001) and albumin and total protein (P < 0.01). Total bilirubin and gamma glutamyl transferase levels were lower in the severely hemolyzed sample (P < 0.001). Differences in mean values for alkaline phosphatase between nonhemolyzed and hemolyzed serum were not significant but did show a downward trend in the hemolyzed sera. The presence and severity of hemolysis must be considered in the interpretation of the serum chemistry values.     

Effects of storage time and freezing temperature on clinical chemical parameters from canine serum and heparinized plasma

To assess changes in 24 blood constituents in frozen serum and heparinized plasma, blood samples were drawn from 10 clinically normal German Shepherd army dogs. The storage characteristics of nine enzymes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, lipase), and 15 metabolites and minerals (albumin, bile acids, bilirubin, calcium, cholesterol, creatinine, fructosamine, glucose, magnesium, phosphate, potassium, protein, sodium, triglycerides, urea) were studied. Parallel samples of serum and heparinized plasma were stored for 90 and 240 days at two different storage temperatures, -200 degrees C and -700 degrees C. Sixteen of the 24 analytes (ALP, ALT, amylase, AST, CK, GGT, GLDH, LDH, bile acids, calcium, cholesterol, creatinine, fructosamine, magnesium, phosphate, urea) showed statistically significant (p < 0.05) changes during the storage period related to storage time, storage temperature, and sample type. Seven of the analytes (amylase, GGT, GLDH, LDH, bile acids, fructosamine, magnesium) showed changes of possible clinical importance with mean differences from baseline larger than 20% for the enzymes and 10% for the metabolites and minerals during the storage periods.     

Age-related changes in plasma biochemical values of farmed emus (Dromaius novaehollandiae)

SUMMARY Blood samples were collected from 40 emus (Dromaius novaehollandiae) of 4 different age groups ranging from 1 week to 14 months. Plasma values of glucose, cholesterol, uric acid, total protein, albumin, creatine kinase, aspartate amino transferase, alkaline phosphatase, calcium, phosphorus and magnesium were measured. Fourteen-month-old birds had lower plasma glucose values and enzyme activities and higher plasma protein values than younger birds. One-week-old birds had higher cholesterol and uric acid values than other age groups. Plasma calcium, phosphorus and magnesium values did not differ across the age profiles sampled.     

Reference serum biochemical values for emus and ostriches.

Reference serum biochemical values were determined in blood samples from 15 male, 18 female, and 4 unsexed emus (Dromaius novaehollandiae) 1 to 48 months old. Serum biochemical values also were obtained for 19 male, 26 female, and 4 unsexed ostriches (Struthio camelus) 1 to 60 months old. Parametric (mean +/- 2 SD) and non-parametric (fifth to 95th percentile) reference ranges and linear trends as influenced by age were determined for enzyme activities and concentrations of glucose, inorganic phosphate, BUN, uric acid, creatinine, triglyceride, cholesterol, total protein, and albumin. Species differences for all analytes, except cholesterol and inorganic phosphate concentrations, were detected. Creatine kinase values in ostriches were higher than those in emus. There were no linear relationships between age and analyte values in emus, and sex did not significantly (P < 0.05) affect the values in emus. Analyte values in ostriches tended to increase with age, but cholesterol, creatine kinase, inorganic phosphate, and alkaline phosphatase concentrations decreased with age. Glucose, triglyceride, gamma-glutamyltransferase, and cholinesterase concentrations in ostriches were not linearly associated with age. Age had a greater effect on the analyte values of female ostriches than it did on male ostriches. Concentrations generally increased with age in female ostriches, except for cholesterol, cholinesterase, inorganic phosphate, and alkaline phosphatase concentrations, which decreased with age.     

Comparison of two dry chemistry analyzers and a wet chemistry analyzer using canine serum

Canine serum was used to compare seven chemistry analytes on two tabletop clinical dry chemistry analyzers, Boehringer's Reflotron and Kodak's Ektachem. Results were compared to those obtained on a wet chemistry reference analyzer, Roche Diagnostic's Cobas Mira. Analytes measured were urea nitrogen (BUN), creatinine, glucose, aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholesterol and bilirubin. Nine to 12 canine sera with values in the low, normal, and high range were evaluated. The correlations were acceptable for all comparisons with correlation coefficients greater than 0.98 for all analytes. Regression analysis resulted in significant differences for both tabletop analyzers when compared to the reference analyzer for cholesterol and bilirubin, and for glucose and AST on the Kodak Ektachem. Differences appeared to result from proportional systematic error occurring at high analyte concentrations.     

Gender Differences–Induced Changes in Serum Hematologic and Biochemical Variables in Mangalarga Marchador Horses After a Marcha Gait Competition

The aim of this study was to investigate whether marcha exercise results in changes in serum hematologic and biochemical variables and whether this condition differs in male and female horses. Thirty-five Mangalarga Marchador horses, 18 males and 17 females were included in the study. Blood samples were obtained before and immediately after a marcha competition. Samples were used for measurement of hematocrit (HCT), hemoglobin concentration (HGB), red blood cell (RBC), total and differential white blood cell (WBC), total protein (TP), albumin (Alb), aspartate aminotransferase (AST), gamma glutamyltransferase (GGT), creatine kinase (CK), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), creatinine (Crea), and urea (BUN) levels. Data were submitted to analysis of variance using the SAS statistical program, and means were compared by the Tukey test (P < .05). In both males and females, RBC, HCT, AST, and GGT activity increased in response to exercise, but there were no significant differences between genders. In females, significant increases in TP, BUN, Crea, CK, and LDH levels were detected after the marcha gait, but no such changes were detected in males. Total WBC concentrations increased in response to the exercise in both, males and females; however, there were no differences between genders. Plasma HGB, Alb, and ALP levels after marcha gait did not differ in both genders. Marcha exercise–induced significant hematologic and biochemical changes in serum of Mangalarga Marchador horses, independently of gender. However, females presented more blood changes after marcha gait. Gender differences should be taken into consideration in experiments with athletic horses.     

Serum constituents analyses in dairy cows: effects of duration and temperature of the storage of clotted blood

We studied the effects of storage time and temperature of clotted whole blood on the amounts of 17 analytes in bovine blood serum. Serum separated after blood was allowed to stand for 2, 4, 6, 12 and 24h at room temperature or on ice. Results obtained for phosphorous, magnesium, urea, cholesterol, beta-hydroxybutyrate (BHBA), triglyceride, albumin, total protein and gamma-glutamyletransferase (GGT) were not influenced by storage at room temperature or on ice for as long as 24h. Duration of the clotted whole blood storage had a significant effect on calcium, glucose concentrations, creatine kinase (CK) and aspartate aminotransferase (AST) activities and temperature had a significant effect on glucose, non-esterified fatty acids, CK and bilirubin concentrations.     

Effect of storage on some constituents of camel serum

SUMMARY Effects of storage at room temperature (23–25°C) and refrigeration (4–5°C) on various biochemical constituents of camel serum were investigated. Albumin, globulin, calcium, phosphorus, cholesterol, aspartate aminotransferase (AST), alkaline phosphatase (AP) and gamma glutamyltransferase (GGT) did not change over 9 days when stored at 4–5°C. At 4–5°C, creatinine, iron and glucose in camel sera remained stable for 6 days; total protein for 7 days; and blood urea nitrogen for 8 days. Decreased activities in creatine kinase (CK), lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) were apparent after 1, 6 and 7 days, respectively. At room temperature, total protein, albumin, globulin, calcium and phosphorus were stable throughout the 9 days. Changes in glucose and iron occurred after 3 days. Stability at room temperature for LDH was 1 day; AST, 3 days; GGT and ALT, 6 days; and AP, 8 days. CK activity had already declined by 4 hours and by 9 days, only 34% activity remained.