Working with rather than against macaques during blood collection.

Training macaques to cooperate during blood collection is a practicable and safe alternative to the traditional procedure implying forced restraint. It takes a cumulative total of about 1 hr to train an adult female or adult male rhesus macaque successfully to present a leg voluntarily and accept venipuncture in the homecage. Cooperative animals do not show the significant cortisol response and defensive reactions that typically occur in animals who are forcibly restrained during this common procedure.

Effect of venipuncture sites on hematologic and clinical biochemical values in desert tortoises (Gopherus agassizii).

Paired blood samples were collected from the postoccipital venous plexus and jugular vein of desert tortoises (Gopherus agassizii) for hematologic and plasma biochemical analyses. Comparison of hematologic values revealed significantly (P < or = 0.05) lower PCV, RBC count, WBC count, and hemoglobin values for samples obtained from the occipital site. When comparisons were made between plasma biochemical values for the 2 sites, significant (P < or = 0.05) differences were measured for: glucose, potassium, chloride, uric acid, calcium, phosphorous, total protein, albumin, globulin, alkaline phosphatase, aspartate transaminase, alanine transaminase, and total cholesterol. Significant differences between hematologic and plasma biochemical values from the occipital region samples vs jugular vein samples were attributed to hemodilution of the occipital region samples with extravascular fluid or lymph or both.

Jugular vein wall repair after intravenous injection in equids.

The evolution of the healing process of venous puncture wounds was studied macroscopically and light-microscopically in order to determine the time of venipuncture. Seven small equids of various ages, all in normal healthy condition, were experimentally injected in the external jugular vein with a physiological Hartmann solution at different times before euthanasia. Two types of needle diameters i.e. 16.5 G and 19 G, were used. A fairly good uniformity in the healing process was seen in the 7 animals. In most cases the lesions induced within 5 weeks before death were macroscopically visible, whereas older perforation sites became indistinct. Light-microscopic study enabled determination of the time lapse after intravenous injection. The needle diameter seemed hereby relatively unimportant. Points of reference were endothelium repair, presence of a fibrin plug containing red and white blood cells, neovascularization and healing of the connective tissue fibers.

Polycythaemia vera in a dog treated by repeated phlebotomies.

Polycythaemia vera (PV) was diagnosed in a dog by demonstration of an increased red cell mass in association with normal arterial oxygen saturation and the absence of conditions known to be associated with secondary polycythaemia. The dog was treated exclusively by repeated phlebotomies and replacement of the removed volume by colloid and crystalloid solutions. It survived for one year and was generally free of signs.

Shipping stress and social status effects on pig performance, plasma cortisol, natural killer cell activity, and leukocyte numbers.

Crossbred pigs were used to evaluate the effects of shipping stress on natural killer (NK) cell activity, leukocyte numbers, plasma cortisol, and BW changes. In the first study, pigs were bled at a commercial farm and, after shipping, resident and shipped pigs were bled again. Plasma cortisol concentrations were not different (P > .10) because of large variation in cortisol concentrations. Furthermore, NK cytotoxicity was nondetectable among all pigs. A second study showed that plasma cortisol concentration rose by approximately 2.6 ng/mL (P = .018) for each minute after pigs were aroused. In the third, more controlled study, pigs were housed in pens of three pigs each. Video recordings were made during the first 24 h pigs were grouped to identify socially dominant, intermediate, and submissive pigs. At time zero (before shipping), resident pigs and those to be shipped had similar plasma cortisol concentrations. However, after the 4-h shipping experience, shipped pigs had elevated (P < .05) plasma cortisol compared with resident control pigs. Shipped pigs lost 5.1% of their BW (P < .05) compared with resident pigs, which gained .02% of their BW. Body weight change during shipping and plasma cortisol were negatively correlated (r = -.34, P = .04), indicating pigs that had greater adrenal response to shipping also lost more weight during shipping. Shipping reduced (P < .05) NK cytotoxicity among pigs of intermediate and submissive social status compared with shipped, dominant pigs. At the end of shipping or control treatments, the correlation between NK cytotoxicity and plasma cortisol was positive (r = .35, P = .036), indicating that pigs with greater cortisol response had greater NK cytotoxicity.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of phlebotomy-induced erythropoietin production in the dog.

Erythropoietin (EPO) production in dogs was studied by reducing red blood cells with phlebotomy. In this study, the hemoglobin reduction rate (delta %Hb) was newly taken into account as the regulating factor for EPO production, and its usefulness to estimate the stimulating intensity to EPO production was examined. As the result, plasma EPO was highly correlated with delta %Hb showing the importance related to regulation of EPO production, in the increasing plasma EPO by different degrees of phlebotomy, in the change of plasma EPO through the anemia progress and recovery period after severe phlebotomy, and in the initial variation of plasma EPO induced by chronic mild phlebotomy. On the other hand, increasing EPO production appeared at least within 6 hr after acute severe phlebotomy, which revealed significantly higher plasma level compared with the mild chronic phlebotomy, suggesting the effect of time leading to red blood cell reduction on EPO production response. Simultaneously, considering an in vivo EPO half life of 8.4 hr calculated from plasma EPO disappearance after bilateral nephrectomy, endogenous plasma EPO accumulation should be taken into consideration in rapidly increasing of delta %Hb.

Practical venipuncture techniques for the ferret.

As the number of ferrets (Mustela putorius furo) used in research and kept as pets continues to rise, so does the need for simple, humane research and diagnostic techniques. We have developed venipuncture methods for the ferret utilizing the jugular and cephalic veins. Using these methods it is possible to repeatedly sample moderate volumes of blood and to perform intravenous injections in both conscious and sedated ferrets.

Effects of chronic blood loss in a marsupial (Monodelphis domestica).

Monodelphis domestica, the gray short-tailed opossum, is used increasingly as an animal model in studies that require repeated blood sampling. Consequently, it is important to establish safe bleeding regimens. We investigated the effects of repeated blood loss on various hematologic values and on different organs in this species. Approximately 2 ml of blood were taken weekly from 20 animals for 13 weeks. The animals were then necropsied; members of an age- and sex-matched control group were bled (2 ml) once and necropsied immediately to obtain baseline data. Ultimately, each animal in the experimental group lost approximately three times its total blood volume. After the first bleeding in the chronically bled group, the red blood cell counts, hemoglobin, and hematocrit values decreased significantly but remained constant thereafter. In another experimental group bled only once, the hematologic values rose to higher than baseline levels after a rest of 2 weeks. Thereafter the values slowly returned to baseline levels. A notable increase in Howell-Jolly bodies occurred in the chronically bled group. Histologically, there was marked erythroid hyperplasia in the bone marrow and extramedullary hematopoiesis in the spleen, but none in the liver. Because there were no obvious detrimental physiologic effects, we conclude that M. domestica is markedly tolerant of chronic blood loss.

Effects of methods used for blood collection on plasma concentrations of luteinising hormone, testosterone, and cortisol in male dogs.

The effects of two putative stressors relative to the collection of blood, namely the environment of the treatment room and the pain associated with venepuncture, on plasma levels of luteinising hormone (LH), testosterone and cortisol were examined in six trained male experimental dogs. Blood samples were collected from the dogs in a treatment room as well as in the kennels (control), and by venepuncture as well as via an indwelling intravenous catheter (control). No significant influence of either stressor on plasma levels of LH, testosterone or cortisol was found. Plasma concentrations of these hormones varied considerably both between and within dogs. Mean (+/- SEM; n = 6) plasma concentrations were 4.3 +/- 1.0 micrograms/l for LH, 4.6 +/- 1.9 nmol/l for testosterone and 68 +/- 10 nmol/l for cortisol. It was concluded that the putative stressors, the environment of the treatment room and the pain associated with venepuncture, did not significantly influence plasma levels of LH, testosterone or cortisol in trained male experimental dogs. This conclusion implies that under the experimental conditions described, the validity of results will not be affected by the method of blood collection used.

Effects of standard anticoagulants and storage procedures on plasma glucose values in seals.

Standard methods for the preservation of vertebrate blood samples for glucose analysis include collecting and storing the blood in evacuated tubes containing sodium fluoride (glycolytic inhibitor) and potassium oxalate (anticoagulant). We found that blood collected from 5 seals by venipuncture and transferred into these tubes had a significantly (P less than 0.05) lower plasma glucose value than blood transferred into tubes containing heparin. In species in which RBC glucose concentration is significantly less than that in the plasma, fluoride and oxalate-induced hemolysis dilutes the plasma with cytoplasm and lowers the measured concentration of glucose in plasma. Therefore, although plasma glucose is used extensively in experimental and clinical analyses, standard techniques for handling the blood may create errors that could confuse comparisons between individuals or species.

Swine models for cardiovascular research: a low stress transport and restraint system for large swine.

A restraint and transport system was developed for handling large swine during cardiovascular research studies. The major design criteria provided for comfortable, low stress restraint of the swine, safety for laboratory personnel and ability to perform a wide variety of hemodynamic and echocardiographic measurements in the standing, supported standing and sedated, or in Panepinto sling positions. A head gate is provided for venipuncture procedures, and an auxiliary feeding and watering front panel can replace the head gate for use of the system as a post-operative "recovery room". Using this system animals weighing 22 to 150 kg can be easily managed.

Effect of repeated phlebotomy on iron status of rhesus monkeys (Macaca mulatta).

Iron status, as determined by hematologic values, serum iron concentration, total iron-binding capacity, and zinc protoporphyrin concentration, was determined in 2 groups of 6 nonpregnant monkeys. Monkeys of groups 1 and 2 had 10 and 5%, respectively, of their blood volume withdrawn per week for up to 10 weeks or until blood hemoglobin concentration was less than or equal to 10 g/dl. A third group of 6 monkeys served as controls. The majority (8/12) of the monkeys became anemic (hemoglobin concentration, less than or equal to 10 g/dl) after approximately 30 to 70% (mean, 49%) of their blood volume was removed. Anemia was accompanied by decrease in serum iron concentration and percentage of transferrin saturation. Microcytosis, hypochromasia, and increased zinc protoporphyrin concentration, all hematologic characteristics of iron deficiency, developed later. The calculated iron stores ranged from 1 to 133 mg, with mean value of 51 mg. Iron-depleted monkeys had mean calculated available iron store of 20.8 mg, whereas iron-replete monkeys had mean available iron store of 114.0 mg. Changes were not observed in monkeys of the control group during the study period. None of the baseline hematologic or biochemical analytes measured were good predictors of iron stores. The diet used at the research center did not provide sufficient iron to prevent iron deficiency in most of the monkeys from which a total amount of 30 to 70% of blood volume at 5 or 10%/week was withdrawn. Studies requiring that much blood may need to be modified to include iron supplementation, reduction of sample volume, or iron replacement after termination of projects.