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[Determination of plasma hemoglobin in fresh plasma for therapeutic use by the TMB (3,3',5,5'-tetramethylbenzidine) method].

The maximum level of plasma hemoglobin in standard fresh frozen plasma before freezing has been mixed at 50 mg/l by french regulations. As no reference method is specified by the standards, we have adapted a technique where pseudo-peroxydasic activity of the hemoglobin is revealed by 3,3'-5,5', tetramethylbenzidine, a non carcinogenic chromogen derivate from benzidine. A 4 points reference scale is plotted for each plasma to be tested, thus reducing eventual interferences between pseudo-peroxydasic activity of the hemoglobin and other plasma proteins. We compared our modified technique to other existing ones: Vanzetti's extraction technique [8] whose main drawbacks are the use of flammable solvents and carcinogenic chromogen. Standefer's technique [10] involving plasma preincubation in H2O2 solution, which reveals protein interferences but is imprecise as it relies on only one reference point and does not show whether chromogen saturation is present. The level fixed by regulations is higher than the mean level indicated by our method, based on 118 samples of fresh plasma obtained by double centrifugation of whole blood collected on CPD as anticoagulant. It is also higher than the level indicated by the same method for plasma samples drawn from continuous or discontinuous flow cytapheresis (IBM 2997; Haemonetics V 50) using ACD - A or ACD - AA 16 as anticoagulant. Our method measures with precision low levels in plasma. Furthermore it is cheap, simple and easy to run in a blood transfusion center.

Benzidines↗

Plasma oxygen during cardiopulmonary bypass: a comparison of blood oxygen levels with oxygen present in plasma lipid.

1. Although not often appreciated, it is a fact that molecular oxygen is more soluble in lipids than in aqueous solution. We have recently developed a method to monitor oxygen within the lipid content of plasma. Monitoring plasma oxygen is one essential element during open heart surgery using a cardiopulmonary bypass pump and oxygenator. Currently oxygen is monitored electrochemically and is based upon monitoring the partial pressure of oxygen in a gas equilibrated with whole blood. 2. To determine the relative importance of lipid-associated oxygen in blood and assess the potential use of such a measurement we present comparisons of changes in oxygen associated with whole blood and lipid content of plasma before, during and after cardiac surgery. 3. In a limited number of patients studied (n = 28), aged between 34 and 86 years, oxygen in lipid increased with decreased extracorporeal blood temperature during cardiopulmonary bypass, increased in proportion to oxygen supplied and appeared to be a better monitor of oxygen than conventional electrochemical systems currently in use. Oxygen associated with whole blood and plasma lipid was markedly below normal on aortic declamping after cardiopulmonary bypass, suggesting an hypoxic episode at this point. Levels of oxygen in the lipid phase of plasma returned to normal presurgical values 6-8 h after surgery. 4. Calculation of the concentration of lipid-associated oxygen present in plasma suggests that plasma lipids contain up to 25% of that typically ascribed to haemoglobin. Thus, we suggest that monitoring lipid-associated oxygen may prove a better alternative to current methods of measuring oxygen status. Furthermore, we suggest that plasma lipid is a hitherto unsuspected pool of circulating oxygen which may play a significant role in tissue oxygen supply.

Adult↗

The use of tri(n-butyl)phosphate detergent mixtures to inactivate hepatitis viruses and human immunodeficiency virus in plasma and plasma's subsequent fractionation.

The treatment of plasma with organic solvent/detergent mixtures at the time of plasma collection or pooling could reduce the exposure of technical staff to infectious viruses and enhance the viral safety of the final product. Treatment of plasma for 4 hours with 2-percent tri(n-butyl)phosphate (TNBP) at 37 degrees C, with 1-percent TNBP and 1-percent polyoxyethylensorbitan monooleate (Tween 80) at 30 degrees C, or with 1-percent TNBP and 1-percent polyoxyethylene ethers, (Triton X-45) at 30 degrees C resulted in the rapid and complete inactivation of greater than or equal to 10(4) tissue culture-infectious doses (TCID50) of vesicular stomatitis and Sindbis viruses, which are used as surrogates. Treatment of plasma with TNBP and TNBP and Tween-80 was shown to inactivate greater than or equal to 10(4) TCID50 of human immunodeficiency virus. TNBP treatment of plasma contaminated with 10(6) chimpanzee-infectious doses (CID50) of hepatitis B virus and 10(5) CID50 of non-A,non-B hepatitis virus prevented the transmission of hepatitis to chimpanzees. Immediately after treatment of plasma with 2-percent TNBP, the recovery of factors VIII, IX, and V and antithrombin III was 80, 90, 40, and 100 percent, respectively. Recovery of all factors was greater than or equal to 90 percent after treatment with TNBP and detergent mixtures. Treated plasma was fractionated by standard techniques into antihemophilic factor and prothrombin complex concentrates, immune globulin, and albumin. Prior treatment with TNBP or TNBP and detergent did not affect the separations of desired proteins. Therefore, it appears possible to inactivate viruses in plasma before the execution of standard fractionation procedures.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Proteins↗

Quantitation of genomic DNA in plasma and serum samples: higher concentrations of genomic DNA found in serum than in plasma.

BACKGROUND: Plasma and serum samples have been used to detect cell-free genomic DNA in serum or plasma in certain pathologic conditions such as systemic lupus erythematosus, pulmonary embolism, and malignancies, as well as in fetal cell chimerisms in maternal serum and/or plasma. In this study, baseline concentrations of cell-free DNA in serum and plasma samples were evaluated for the study of posttransfusion chimerism. STUDY DESIGN AND METHODS: DNA was extracted from fresh or stored (4 degrees C for 1-6 days) normal donor serum or plasma samples (ACD; EDTA) by using reagents from an HIV assay kit. After incubation and washing of samples, purified DNA was amplified with HLA DQ-alpha primers (GH26 and 27) or human Y-chromosome primers (SA and SD) to quantitate the concentration of genomic DNA. RESULTS: Fresh serum samples had concentrations of cell-free DNA that were about 20-fold higher than the concentrations in fresh plasma samples. The concentration of cell-free genomic DNA in serum samples increased daily, to a level more than 100 times baseline after clotted blood tubes were stored at 4 degrees C for 4 to 5 days. There was a small increase in cell-free plasma DNA in stored ACD whole blood samples. Male WBCs, spiked into fresh nonanticoagulated female blood, were lysed during the process of clotting, with male DNA liberated into the serum samples. CONCLUSION: Most cell-free DNA in serum samples is generated during the process of clotting in the original collection tube. The concentration of cell-free genomic DNA in fresh plasma is probably the same as that in circulation. Consequently, while serum samples should not be used to monitor the concentration of cell-free DNA in a patient's circulation, serum collected from sample tubes containing clots (i.e., without anticoagulant), 3 to 5 days after the date of phlebotomy, could be useful as a source of DNA with which to screen for posttransfusion microchimerism.

Blood Donors↗

Evaluation of plasma retinol-binding protein as a surrogate measure for plasma retinol concentrations.

Plasma retinol-binding protein (RBP) concentrations have been suggested as surrogate indicators for plasma retinol concentrations in the assessment of vitamin A status in less technologically developed settings. Plasma RBP was measured by radial immunodiffusion and plasma retinol by high performance liquid chromatography in a cross-sectional study of 900 pregnant women at the Queen Elizabeth Central Hospital, Blantyre, Malawi. The Spearman correlation coefficient between plasma RBP and retinol concentrations was 0.95 (p<0.0001). By linear regression, 0.70 micromol l(-1) retinol was equivalent to 21.1 mg l(-1) RBP. With these cut-off points for defining vitamin A deficiency, there was high concordance between categorical descriptions of deficiency using retinol and RBP by chi-square analysis (p<0.001). Measurement of plasma RBP by radial immunodiffusion is simple, inexpensive, and does not require expensive instrumentation. Plasma RBP concentrations measured by radial immunodiffusion are highly correlated with plasma retinol and can be used as a simple surrogate measure for vitamin A concentrations in large field studies.

Adult↗

Coagulation factor content of solvent/detergent plasma compared with fresh frozen plasma.

Solvent/detergent (S/D) plasma is being increasingly widely used in clinical practice, as it carries significantly lower risk of lipid-enveloped viral transmission than standard fresh frozen plasma (FFP). However, previous reports have suggested that S/D processing might influence the coagulation factor content of plasma. We have investigated this question by measuring procoagulant factors (fibrinogen, factor V and factor VIII), anticoagulant factors (protein C and protein S) and routine coagulation screening tests (prothrombin time and activated partial thromboplastin time) in 48 single-donor units of FFP, and in 16 units of S/D plasma (Octaplas). All routine coagulation screening tests, factor VII and protein C levels were within the normal reference range for both S/D plasma and FFP. However, we found significant reductions in factor V (31%), factor VIII (28%) and protein S (50%) in S/D plasma. The observed quantitative differences in coagulation factor levels may be further exacerbated by the lower volume of solvent detergent plasma units (200 ml) compared with units of standard fresh frozen plasma (250 ml). These findings are of potential clinical significance, particularly in those patients with liver disease, constitutional factor V deficiency and congenital or acquired protein S deficiency.

Blood Coagulation Factors↗

Leukaemic peripheral blood plasma and bone marrow plasma: comparison of influence on lymphocyte proliferation.

Peripheral blood plasma from some children with untreated acute lymphoblastic leukaemia (ALL) exerted an inhibitory effect in vitro on phytohaemagglutinin-induced lymphocyte transformation of normal peripheral blood lymphocytes. This occurred at concentrations beyond that required for optimal response as judged by reduction of blast cell formation and tritiated thymidine and tritiated uridine incorporation into DNA and RNA, respectively. In contrast, bone marrow plasma from these patients was non-inhibitory or contained significantly less inhibitory activity. Bone marrow plasma from the majority of healthy controls was superior to their peripheral blood plasma in enhancing phytohaemagglutinin-induced mitogenesis. The difference between an individual's bone marrow- and peripheral blood-derived plasma in enhancing proliferation of patient and healthy control cells was significantly greater amongst the patients than the healthy control group; this was attributed mainly to the increased inhibitory activity of ALL peripheral blood plasma compared with normal plasma. Medium conditioned by phytohaemagglutinin-stimulated normal peripheral blood lymphocytes was effective in neutralizing the inhibitory activity of ALL peripheral blood plasma. Taken together, these in vitro results are at least suggestive that in vivo, in healthy subjects, the rapidly proliferating cells in the bone marrow and the 'resting' blood cells in the circulation may be under the influence of a fine balance of different types and/or levels of humoral growth stimulatory and inhibitory factors and that in ALL an unstable balance of these factors exists. The decreased proliferation of circulating blast cells compared with bone marrow blasts in ALL may be attributed, at least in part, to exposure to the different levels of inhibitor(s) in the circulation and bone marrow as demonstrated in vitro by our results.

Bone Marrow↗

Effect of lipoprotein concentration and lecithin: cholesterol acyltransferase activity on cholesterol esterification in human plasma after plasma exchange.

The rate of cholesterol esterification in plasma, plasma lecithin cholesterol acyltransferase (LCAT) activity and plasma lipoprotein levels have been measured in five subjects who underwent therapeutic plasma exchange to reduce their plasma cholesterol concentration. In the week following the exchange the cholesterol esterification rate and the plasma triglyceride concentration returned rapidly in parallel to pre-exchange levels, while high density lipoprotein (HDL) cholesterol and LCAT activity returned to normal more slowly but also in parallel. The data suggest that the rate-limiting factor for cholesterol esterification in plasma is unlikely to be solely the enzyme levels, but is probably a combination of factors, including the enzyme level and either substrate availabiltiy or product removal. Plasma very low density lipoprotein (VLDL) may either provide substrates for the reaction or provide a means of removing one of the products from the site of reaction.

Adult↗

Selective plasma component removal: alternatives to plasma exchange.

Plasma exchange, the unselective removal of all plasma constituents, has been applied, with varying degrees of success, to an increasing number of disorders. Because of the high cost of plasma exchange, largely the result of expensive homologous replacement solution, and other reasons, alternatives have been sought. Selective removal of plasma components permits the use of autologous plasma as replacement which is less costly, more physiologic, and in some cases, a more efficient alternative to plasma exchange. The selective removal of plasma components whose presence is associated with a disease process can be accomplished presently by physical or chemical means with either on-line of off-line systems. A state-of-the-art review of currently available alternatives to plasma exchange is described. This includes hemoperfusion and plasmaperfusion over affinity columns or filters with immobilized sorbents. On-line separation of macromolecules by membrane filtration and cryoprecipitation as well as off-line chemical precipitation and cryoprecipitation are also described. The potential and limitations of the techniques are emphasized.

Agglutinins↗

Plasma expansion does not precipitate the fall in plasma vasopressin in humans drinking isotonic fluids.

In a group of healthy humans, plasma vasopressin (AVP) levels fell on drinking either Tyrode or mannitol solutions isosmotic with plasma. Both the timing and magnitude of the fall were appropriate to account for the transient diuresis which followed the drinking. Although plasma expansion follows drinking Tyrode solution it occurred too late to account for the fall in plasma AVP. It was also too small to inhibit AVP secretion. Even though plasma volume tended to contract on drinking isosmotic mannitol solution a fall in plasma AVP and a diuresis occurred, similar to those found after drinking Tyrode solution. These findings appear to eliminate plasma volume expansion as the stimulus for the fall in plasma AVP and the associated diuresis on drinking isotonic fluids. In a further group of human subjects, bypassing the oropharynx by intragastric infusion resulted in a slower onset of diuresis after a water load. We suggest that receptors, as yet undefined, in the upper gastrointestinal tract contribute to the early stages of a water diuresis and account for the apparently inappropriate transient diuresis which follows the drinking of isotonic fluids.

Adult↗

Adverse reactions to plasma and plasma components.

Since the early days of plasma fractionation, plasma and derivatives have been identified as being responsible for a spectrum of mild to life-threatening reactions, but it is increasingly being recognised that plasma and its derivatives may be responsible for a wider range of reactions in recipients and more frequently than is commonly appreciated by clinicians. These reactions may be related to immunological differences between the donor and the recipient, either with the component being antigenic to the recipient or the plasma containing an antibody reacting with a recipient antigen. Secondly, there may be physicochemical characteristics of the plasma component such as temperature, additives, alterations due to preparative process and accumulation of metabolites or cellular release products on storage. All of these may in some patients be responsible for clinically significant reactions. This review presents a broad overview of the clinical presentations of recipient reactions due to plasma or plasma components and their underlying mechanisms. Unless there is an immediate or acute fulminant reaction to the infusion of plasma component, adverse effects of these blood products frequently go unrecognised and untreated. Some of the reactions, if undiagnosed, may have a significant effect on the outcome of the patient. There are also various preventative measures which may be taken in the preparation of blood products or their administration to minimise their potential adverse effects.

Blood Component Transfusion↗

Generation of migratory antigen-specific plasma blasts and mobilization of resident plasma cells in a secondary immune response.

Maintenance of protective humoral immunity depends on the generation and survival of antibody-secreting cells. The bone marrow provides niches for long-term survival of plasma cells generated in the course of systemic immune responses in secondary lymphoid organs. Here, we have analyzed migratory human plasma blasts and plasma cells after secondary vaccination with tetanus toxin. On days 6 and 7 after immunization, CD19(+)/CD27(high)/intracellular immunoglobulin G(high) (IgG(high))/HLA-DR(high)/CD38(high)/CD20(-)/CD95(+) tetanus toxin-specific antibody-secreting plasma blasts were released in large numbers from the secondary lymphoid organs into the blood. These cells show chemotactic responsiveness toward ligands for CXCR3 and CXCR4, probably guiding them to the bone marrow or inflamed tissue. At the same time, a population of CD19(+)/CD27(high)/intracellular IgG(high)/HLA-DR(low)/CD38(+)/CD20(-)/CD95(+) cells appeared in the blood in large numbers. These cells, with the phenotype of long-lived plasma cells, secreted antibodies of unknown specificity, not tetanus toxoid. The appearance of these plasma cells in the blood indicates successful competition for survival niches in the bone marrow between newly generated plasma blasts and resident plasma cells as a fundamental mechanism for the establishment of humoral memory and its plasticity.

Adult↗

Plasma volume substitution does not inhibit plasma noradrenaline and muscle nerve sympathetic responses to insulin-induced hypoglycaemia in healthy humans.

Microelectrode recordings of muscle nerve sympathetic activity and measurements of venous plasma noradrenaline have indicated increased sympathetic outflow during acute hypoglycaemia. Plasma volume reduction during hypoglycaemia, as evidenced by increasing peripheral venous haematocrit might underly the sympathetic activation. To study the effect of prevention of plasma volume reduction during hypoglycaemia, saline containing albumin was infused intravenously in healthy adult volunteers during hypoglycaemia. Hypoglycaemia was induced by an intravenous injection of soluble insulin in a dose of 0.15 IU/kg body weight. Peripheral venous plasma noradrenaline concentrations were identical in experiments without and with plasma volume substitution. Muscle nerve sympathetic activity increased to the same extent during hypoglycaemia with and without plasma volume substitution. It is concluded that increased plasma noradrenaline concentrations and enhanced muscle nerve sympathetic activity during insulin-induced hypoglycaemia in man are not consequences of plasma volume reduction.

Adult↗

Monitoring oral anticoagulant treatment from plasma stored for up to 48 hours and frozen plasma.

BACKGROUND AND OBJECTIVE: The number of patients being referred for lifelong anticoagulant treatment has increased markedly in the last years. The prothrombin time test is sometimes difficult to perform the same day as sample collection. The aim of this study is to determine International Normalized Ratio (INR) and vitamin-K dependent factor levels of frozen plasma and plasma stored for up 48 hours. DESIGN AND METHODS: The INR of 84 patients receiving acenocoumarol were determined fresh (0 hours), on samples stored between 2 degrees C and 8 degrees C for 24 hours and 48 hours, and on frozen samples (-40 degrees C) using 4 different thromboplastin reagents (Thromboplastin IS; Thromborel; Simplastin; and Thromboplastin D+G). In addition, factors II, VII, IX, X were determined in 34 of these patients in all these situations. We used the interclass correlation coefficient to compare the results obtained at 0 hours and the results obtained in the subsequent measurements. Both measurement and proportional errors were also estimated by linear regression analysis. RESULTS: The correlation coefficient of the INR between fresh and frozen plasma was 0.98, 0.98, 0.92 and 0.97 for IS, Thromborel, Simplastin and D+G respectively. The correlation between 0 and 24 hours was 0. 98, 0.91, 0.95 and 0.85 for IS, Thromborel, Simplastin and D+G respectively. By 48 hours although IS still had r=0.94, Thromborel, Simplastin, and D+G had r=0.55, r=0.50 and r=0.81, respectively. By 24 hours in stored plasma and in frozen plasma the activity of vitamin-K dependent factors was slightly reduced (r=0.97 at 24h/r=0. 94 with frozen plasma for factor II, r=0.92/0.96 for factor VII, r=0. 83/0.98 for factor IX, and r=0.98/0.95 for factor X). By 48 hours however, significant reductions were noted in the activity of these factors (r=0.94 for factor II, r=0.88 for factor VII, r=0.70 for factor IX, and r=0.98 for factor X). INTERPRETATION AND CONCLUSIONS: The INR can be reliable determined in frozen plasma and in plasma stored at 2-8 degrees C for up to 24 hours.

Administration, Oral↗

Survival of foals with experimentally induced Rhodococcus equi infection given either hyperimmune plasma containing R. equi antibody or normal equine plasma.

The purpose of this study was to determine if colostrum-deprived foals with experimentally induced Rhodococcus equi pneumonia have a decreased severity of the disease and decreased mortality rate when given hyperimmune (HI) R. equi antibody plasma (R. equi titer at least 100 % and virulence-associated protein A [VapA] at least 10000) prophylactically versus when given normal equine plasma (R. equi titer less than 20 % and VapA less than 160). Sixteen colostrum-deprived foals (R. equi titer less than 5 %) each received normal equine plasma in the first 24 hours of life (R. equi titer less than 20 %). At 14 days of age, six foals were given normal equine plasma and 10 foals were given HI plasma. All foals were subsequently infected intrabronchially with a pathogenic strain of R. equi (2.5 x 10 sup 8; organisms) at 21 days of age. Repeated physical examinations, weight measurements, complete blood cell counts, fibrinogen measurements, and thoracic radiographs (ventrodorsal and lateral) were performed to help determine the severity of the disease. Foals given HI plasma had significantly higher R. equi ELISA titers (42.4 %) than those given normal plasma (20.9 %) on the day of experimental infection. Mortality rates and severity of disease were statistically similar (P >.05) for the groups. Although none of the foals was treated with antibiotics, several with severe R. equi pneumonia recovered. Either HI or normal equine plasma administered to foals in the first few weeks of life caused no adverse effects and may be protective against R. equi, although the exact constituent responsible for protection is undetermined and requires further investigation.

Actinomycetales Infections↗

Modulation by chronic nifedipine of plasma atrial natriuretic peptide, cell Na+ transport and plasma volume in rats with renal hypertension.

The influence of a 6-week treatment of female Wistar rats with one-kidney, one clip (1-K, 1 C) renal hypertension, with the calcium antagonist nifedipine on plasma volume, red cell Na+ handling and plasma atrial natriuretic peptide immunoreactivity (ANP-IR) was studied. Measurements were performed at 2 and 6 weeks after surgery. In 1-K, 1 C rats plasma volume was increased and red cell Na+ pump activity was reduced only after 2 weeks. Blood pressure, heart weights and plasma ANP were massively increased after both 2 and 6 weeks. 1-K, 1 C-rats treated with nifedipine had normal plasma volume, plasma ANP, and red cell Na+ pump activity in comparison with sham-operated rats. Increases in blood pressure and heart weights were attenuated. It is concluded that 1-K, 1 C hypertension in the rat is associated with cardiomegaly, rise in plasma ANP, initial hypervolemia and depression of the membrane Na+ pump. Nifedipine prevents the occurrence of hypervolemia and secondary phenomena such as rises in plasma ANP and cardiomegaly. This may play an important contributory role in the prevention of pathological sequelae.

Animals↗

[Improved the accuracy of 99mTc-MAG3 plasma clearance method: the problem of the calculated plasma volume and its modification].

The 99mTc-MAG3 plasma clearance method (MPC method), reported by Oriuchi et al., is a simple and useful count-based gamma camera method for calculating the 99mTc-MAG3 plasma clearance (CLMAG). However, a discrepancy of CLMAG calculated by MPC method (MPC-CLMAG) from the tubular extraction rate (TER) calculated by Russell's single-sample clearance determination (Russell-TER) was noted. The calculated plasma volume is assumed to be the cause. Since the plasma volume is reported to have a linear correlation with body surface area, Dissmann's formula was applied to calculate the plasma volume. Then Dissmann's formula was replaced by Ogawa's formula in the MPC method, and the procedure was then called the modified MPC method. The CLMAG were obtained using MPC method, modified MPC method and the TER was obtained Russell's method in 95 patients with urological disorders. Then the MPC-CLMAG and modified MPC-CLMAG were compared with Russell-TER. Comparison of the MPC-CLMAG with the Russell-TER demonstrated a coefficient of correlation of 0.82, but dissociation of the slope of regression lines was found between males and females. The modified MPC-CLMAG improved the coefficient of correlation to 0.92, and diminished the dissociation of the slope of regression lines between males and females. We verified that the dissociation was due to the plasma volume calculated by Ogawa's formula. Ogawa's formula included hematocrit, body weight, body height and different coefficients for gender. The plasma volume calculated by Ogawa's formula were lower in males and higher in females than that calculated by Dissmann's formula. And marked discrepancy in the plasma volume in patients with a body surface area below 0.5 m2 was observed. So the MPC method might become more accurate by substituting Dissmann's formula for Ogawa's formula resolting in a method that is applicable to both males and females, children and adults in clinical use.

Adolescent↗

Plasma catecholamines and plasma corticosterone following restraint stress in juvenile alligators.

Ten juvenile alligators, mean body mass 793 g, hatched from artificially incubated eggs and raised under controlled conditions, were held out of water with their jaws held closed for 48 hr. An initial blood sample was taken and further samples collected at 1, 2, 4, 8, 24, and 48 hr. Epinephrine, norepinephrine, and dopamine were measured in plasma aliquots of 1.5 ml using high pressure liquid chromatography with electrochemical detection. Corticosterone was measured by radioimmunoassay. Plasma glucose was measured using the Trinder method and plasma calcium, cholesterol, and triglycerides were measured in an autoanalyzer. Epinephrine was about 4 ng/ml at the initial bleed, but declined steadily to < 0.4 ng/ml by 24 hr. Norepinephrine was also about 4 ng/ml at the initial bleed, but rose to over 8 ng/ml at 1 hr, and then declined to < 0.2 ng/ml at 24 hr. A second, but smaller increase in plasma norepinephrine was seen at 48 hr. Plasma dopamine was low at the initial bleed (< 0.7 ng/ml), rose to over 8 ng/ml at 1 hr, then declined to < 0.2 ng/ml. Plasma corticosterone rose progressively for the first 4 hr, declined at 8 hr and 24 hr, then rose again at 48 hr. Plasma glucose rose significantly by 24 hr and remained elevated for 48 hr. Plasma calcium increased at 1, 2, and 4 hr then returned to levels not significantly different from the initial sample at 24 and 48 hr. The white blood cells showed changes indicating immune system suppression. By the end of the treatment the hetorophil/lymphocyte ratio increased to 4.7. These results suggest that handling alligators, taking multiple blood samples, and keeping them restrained for more than 8 hr is a severe stress to the animals.

Alligators and Crocodiles↗