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.     

Mitogen-induced lymphocyte proliferation in loggerhead sea turtles: comparison of methods and effects of gender, plasma testosterone concentration, and body condition on immunity

A fully functioning immune system is vital to the survival of threatened and endangered sea turtles. Immunological protection against diseases in any organism can be reduced by a number of natural and anthropogenic factors, such as seasonal changes, malnutrition, disease states, and contaminant exposure. These factors are even more critical when they occur in endangered species or populations. To identify alterations in the immunological health of loggerhead sea turtles (Caretta caretta), the mitogen-induced lymphocyte proliferation (LP) assay was developed using peripheral blood leukocytes (PBLs). Collection and culture conditions were optimized for this assay using non-lethal blood samples collected from free-ranging turtles along the southeastern US coast. During the collection, two anticoagulants (sodium heparin and lithium heparin) were compared to determine effects of different ions on assay results. Optimal culture conditions were established for loggerhead PBLs while two different methods of measuring LP were compared: (1) the traditional radioactive (3)H-thymidine assay and (2) a non-radioactive, colorimetric method utilizing 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium (MTT). The results indicate that the (3)H-thymidine and the non-radioactive MTT methods did not correlate with each other and that the use of heparin type did not influence the results of the LP assay. Lastly, using these optimized methods, we investigated the effect of gender, plasma testosterone concentration, and body condition on LP in loggerhead turtles and found that none of the parameters largely influenced LP.     

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.     

Effects of anticoagulant and autoanalyzer on blood biochemical values of loggerhead sea turtles (Caretta caretta).

We evaluated the effect of anticoagulant (lithium heparin, sodium heparin, or none) and type of autoanalyzer on selected blood biochemical values of the loggerhead sea turtle (Caretta caretta). More differences were observed between the analytes in serum and those in the 2 types of plasma than were observed between the 2 types of plasma. Differences in electrolyte concentrations were not significant when plasma from sodium-heparinized blood was compared with plasma from lithium-heparinized blood. Serum is not recommended for reptilian studies because clot formation is unpredictable and because the time required for clotting may allow substantial changes in the chemical composition of the sample. For most determinants, values varied more between the 2 types of autoanalyzers than among the 3 anticoagulant treatments. These sources of variation must be considered when performing comparative studies.     

Successive changes of hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta).

Hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta) from the ages of 1 mo to 3 yr were obtained to establish baseline values. Five clinically normal loggerhead turtles were selected from the same clutch and raised in an indoor artificial nesting beach. Blood samples were successively collected and examined for various blood characteristics for a maximum total of 15 times. Hematologic characteristics, including packed cell volume, white blood cell counts, and white blood cell differentials; and plasma chemistry values, including total bilirubin, total protein, albumin, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, gamma-glutamic transpeptidase, creatinine, blood urea nitrogen, uric acid, alkaline phosphatase, amylase, triglyceride, total cholesterol, ionized sodium, ionized potassium and ionized chlorine, were measured. These results were used to establish a hematology and blood chemistry baseline for captive juvenile loggerhead turtles and will aid in their medical management.     

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.     

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 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.     

Plasma concentrations of enrofloxacin after single-dose oral administration in loggerhead sea turtles (Caretta caretta).

Plasma concentrations and pharmacokinetics of enrofloxacin were determined in 12 loggerhead sea turtles (Caretta caretta) after oral administration. Six turtles in group 1 and group 2 received enrofloxacin at 10 mg/kg and 20 mg/kg of body weight, respectively. Blood was collected from the cervical sinus before administration and at timed intervals up to 168 hr following administration. Plasma concentrations of enrofloxacin were determined using a microbiologic assay. The mean peak plasma concentration (Cmax) was 4.07 microg/ml and 21.30 microg/ml for groups 1 and 2, respectively. Plasma levels were detectable at 168 hr postadministration, with mean values of 0.380 microg/ml for group I and 2.769 microg/ml for group 2. The mean elimination half-life for enrofloxacin was 37.80 hr for group I and 54.42 hr for group 2. These findings indicated that enrofloxacin is absorbed following oral administration in loggerhead sea turtles, and blood levels are maintained up to 168 hr following administration.     

Evaluation of hematology and serum biochemistry of cold-stunned green sea turtles (Chelonia mydas) in North Carolina, U.S.A

Hypothermia or cold-stunning is a condition in which the body temperature of an animal decreases below normal physiologic range and which has been linked to severe morbidity in sea turtles. Reports have focused on the physiologic changes caused by cold-stunning in Kemp's Ridley sea turtles (Lepidochelys kempii) and loggerhead sea turtles (Caretta caretta), but few have evaluated the green sea turtle (Chelonia mydas). This study evaluated hematologic and serum biochemical profiles of cold-stunned green sea turtles in North Carolina, USA. When compared with healthy, free-ranging juvenile green turtles from the same region, cold-stunned turtles exhibited hypoglycemia, hypocalcemia (both total and ionized calcium), hyponatremia, hypokalemia, hypoproteinemia, hypoalbuminemia, hyperphosphatemia, and elevations in uric acid and blood urea nitrogen. These findings contrast with some previously reported changes in cold-stunned Kemp's Ridley and loggerhead sea turtles. These results emphasize the importance of basing therapeutic regimens on biochemical analyses in cold-stunned sea turtles.     

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.     

Temporal effects of 3 commonly used anticoagulants on hematologic and biochemical variables in blood samples from macaws and Burmese pythons

BACKGROUND: Few studies have been done to evaluate anticoagulants for use with blood samples from birds and reptiles. Heparin currently is the most commonly used anticoagulant in practice, but may adversely affect blood cell staining and quantitation. OBJECTIVE: The purpose of this study was to evaluate the effects of lithium heparin, K3-EDTA, and sodium citrate, with and without the addition of albumin, on hematologic variables in macaw (Ara sp) and python (Python molurus bivittatus) blood samples. METHODS: Blood samples from 10 macaws and 10 Burmese pythons were collected in heparin-coated syringes and placed into tubes containing either lithium heparin, K3-EDTA, or sodium citrate with and without the addition of 0.25 mL of a 22% bovine serum albumin solution. Cell lysis was determined by counting the number of lysed cells/200 WBCs in Wright's-Giemsa-stained blood smears and by qualitative evaluation of pink plasma in microhematocrit tubes. A CBC was done after 3, 12, and 24 hours of storage at 4 degrees C in anticoagulant-containing tubes and results were compared with those obtained at 0 hour for the heparin-coated syringe sample. A biochemical panel also was done at each time point in similarly stored lithium-heparin samples. RESULTS: Hemolysis was significantly increased in citrated samples from both macaws and pythons beginning at 12 hours. At 24 hours, 19 of 30 (63%) macaw samples in all anticoagulants had >100 lysed cells/200 WBCs. There were no significant differences in hematologic values in samples from pythons collected in heparin or EDTA at any time point. No significant differences were found in the number of lysed cells or in other hematologic data in samples with albumin. Glucose concentration decreased and potassium concentration increased significantly over time in heparinized blood samples. CONCLUSIONS: Based on the results of this study, whole blood samples anticoagulated with lithium heparin or EDTA should be evaluated within 12 hours (macaws) or 24 hours (pythons) of collection and stored at 4 degrees C for best results. Citrate should be avoided as it may result in increased cell lysis. The addition of albumin does not prevent cell lysis.     

Relationship between barnacle epibiotic load and hematologic parameters in loggerhead sea turtles (Caretta caretta), a comparison between migratory and residential animals in Pamlico Sound, North Carolina.

Health status of a total of 57 loggerhead sea turtles (Caretta caretta; 42 migratory and 15 residential turtles) was analyzed using body condition and hematologic parameters. A subset of 18 juvenile migratory loggerhead sea turtles in the fall of 1997 and 15 residential turtles in the summer of 2000 were analyzed for barnacle epibiota. The migratory group had significantly higher red blood cell counts and percent heterophils and significantly lower percent lymphocyte and absolute eosinophil counts, as well as significantly lower plasma concentrations of calcium, sodium, chloride, potassium, glucose, alkaline phosphatase, and anion gap. Many of these variations may be because of physiology of migration. A positive association between turtle weight and hematocrit was detected and may be because of larger turtles diving for longer periods of time. There were no significant differences of epibiota load, health of the turtles, or condition index between turtles captured during the two events.     

Morphologic and cytochemical characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

A morphologic classification based on the cytochemical characteristics of blood cells of 35 juvenile loggerhead sea turtles (Caretta caretta) is described. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff, and toluidine blue. The morphologic characteristics of erythrocytes were similar to those reported in green turtles. Six types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes, monocytes and thrombocytes. Except for the basophils, the rest of the white blood cells from loggerhead turtles had different cytochemical characteristics compared to blood cells from other sea turtle species. The leukocyte differential count was different from that reported for other sea turtle species. Heterophils were the most numerous leukocytes from these loggerhead turtles, followed by lymphocytes, eosinophils, monocytes and basophils. This paper provides a morphologic classification of blood cells of loggerhead sea turtles that is useful for veterinary surgeons involved in sea turtle conservation.     

Comparison of serum and plasma for determination of blood biochemical values in Malaysian flying foxes (Pteropus vampyrus).

The effects of the anticoagulant sodium heparin and time of centrifugation on 20 biochemical analytes in the blood of Malaysian flying foxes (Pteropus vampyrus) were evaluated. Paired plasma and serum samples were centrifuged at 1 hr and 6 hr postcollection. Heparinization and time of centrifugation did not significantly affect albumin, cholesterol, triglycerides, amylase, blood urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransaminase, alkaline phosphatase, calcium, sodium, and total carbon dioxide levels. Plasma was associated with higher globulin and lower potassium values. Glucose and chloride levels decreased significantly over time, whereas phosphorus levels increased. Serum creatine kinase activity at 6 hr postcollection was significantly higher than the other creatine kinase means. Sodium levels were not significantly affected by sodium heparin as used as an anticoagulant in this study.     

Ultrastructural characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

Ultrastructural characteristics of erythrocytes, heterophils, eosinophils, lymphocytes, monocytes and thrombocytes of the loggerhead sea turtle (Caretta caretta) were evaluated, using blood samples from 15 healthy juvenile animals. Except for the eosinophils, the rest of the white blood cells from loggerhead turtles had similar ultrastructural characteristics compared with blood cells from other sea turtle species. Eosinophils from loggerhead turtles were homogeneous in size, and no crystalline structures were observed within the granules. This paper provides an ultrastructural characterization of blood cells of loggerhead sea turtles, as a reference for future haematological studies of this species.     

Pharmacokinetics of ticarcillin in the loggerhead sea turtle (Caretta caretta) after single intravenous and intramuscular injections.

Three captive loggerhead sea turtles, Caretta caretta, were used in four trials, one i.v. and three i.m., to determine the pharmacokinetic properties of a single dose of ticarcillin. For the i.v. study, each turtle received a single 50 mg/kg dose and blood samples were collected at 0, 0.5, 1, 2, 4, 6, 8, and 12 hr and at 1, 1.5, 2, 2.5, 3, 4, 6, 8, 10, and 14 days after administration. For the i.m. study, each turtle received one of three dosages (25, 50, or 100 mg/kg) in a randomized complete block design and blood samples were collected at the same time intervals. Each trial was separated by a minimum of 28 days to allow for complete drug clearance. Drug concentration in plasma was determined by a validated liquid chromatography-mass spectrometry assay. For the i.v. study, the elimination half-life was 5.0 hr. The apparent volume of distribution and plasma clearance were 0.17 L/kg and 0.0218 L/hr/kg, respectively. For the i.m. study, mean time to maximum plasma concentrations ranged from 1.7 ( +/- 0.58) hr in the 50 mg/kg group to 3.7 (+/- 2.5) hr in the 100 mg/kg group. Mean bioavailability ranged from 45% ( +/- 15%) in the 50 mg/kg group to 58% (+/- 12%) in the 100 mg/kg group, and the mean residence time ranged from 7.5 ( +/- 2.6) hr in the 25 mg/kg group to 16 (+/- 6.8) hr in the 100 mg/kg group. Two turtles had slight alanine aminotransferase elevations that were not clinically apparent at two different dosages, but otherwise, blood chemistries were unaffected. Possible i.m. dosage regimens for loggerhead sea turtles are 50 mg/kg q24 hr or 100 mg/kg q48 hr. Liver enzymes should be monitored during treatment.     

Relationship Between Fertility at Foal Heat and Blood Biochemistry Parameters Monitored During the Peripartum Period in Thoroughbred Mares

This study aimed to examine fertility at foal heat and its relevance to body condition score (BCS) and blood nutritional metabolites in Thoroughbred mares. Thoroughbred mares foaled from 2006 to 2009 were included and classified into two groups: group C (conception; n = 34), which included mares that conceived during foal heat (within 3 weeks after foaling), and group NC (nonconception; n = 39), which included mares that did not conceive despite mating during their foal heat. BCS and blood samples were obtained 1 month before the expected foaling date and 1, 2, 3, and 4 weeks after foaling. Total protein (TP), albumin (Alb), blood urea nitrogen (BUN), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), total cholesterol (T-Cho), triglyceride (TG), nonesterified fatty acid (NEFA), calcium (Ca), inorganic phosphate (iP), and magnesium (Mg) levels were measured using an automatic clinical chemistry analyzer. Repeated measures analysis of variance (ANOVA) and Student t-test were used to examine the differences between the two groups. No significant differences were observed between the two groups in any of the above-mentioned parameters at 1 month before the expected foaling date (Student t-test). Furthermore, no significant differences were observed between the two groups in serum levels of TP, Alb, AST, GGT, T-Cho, NEFA, Ca, and Mg and BCS at postpartum periods (repeated measures ANOVA). Serum TG, BUN, and iP levels remained lower in group NC than in group C after foaling (P < .05, repeated measures ANOVA). Although the mechanism by which these nutritional factors affect a decline in reproductive performance remains unclear, our results suggest that blood biochemical tests can detect potential imbalances in nutrition and metabolism, even if there is no difference in BCS.