Blood Collection Techniques Research Articles

Sampling of adult boars during early infection with porcine reproductive and respiratory syndrome virus for testing by polymerase chain reaction using a new blood collection technique (blood-swab method)

Objectives: To evaluate the effectiveness of a skin puncture technique (blood-swab method) to collect blood for detection of porcine reproductive and respiratory syndrome virus (PRRSV) by polymerase chain reaction (PCR) in experimentally infected boars, and to examine the effect of sampling method (blood swab, serum, semen, or mouth swab) and rectal temperature on the ability to detect PRRSV in boars during the first 6 days after experimental infection. Materials and methods: Twenty-one mature boars were inoculated with PRRSV variant MNB04. Serum samples, blood swabs, mouth swabs, and semen samples were collected from two groups of 10 boars on a rotating basis, one group every 24 hours for 6 days. Rectal temperatures were recorded for all boars every 24 hours. Using the blood-swab method, a vein (usually the auricular vein) was punctured with a needle and blood was collected with a polyester swab. The swab was then put into a tube containing 1 mL of saline solution, which was tested for PRRSV virus by PCR. Results: Sixty of 61 samples were PCR-positive using the blood-swab method compared with 61 of 61 PCR-positive using traditional serum collection methods. Testing of both serum and blood swabs detected PRRS-positive boars earlier and with higher frequency than testing semen samples (27 of 60 positive) or mouth-swab samples (19 of 61 positive). Implications: The blood-swab method is a reliable alternative to traditional venipuncture during the first 6 days after infection with PRRSV, and PCR-testing blood swabs is more sensitive than testing semen or mouth swabs.

Acute normovolemic hemodilution in orthopedic surgery

SUMMARYAcute normovolemic hemodilution (ANH) is an autologous blood collection technique that involves the removal of whole blood from a patient on the day of surgery shortly before an anticipated significant surgical blood loss. ANH is one of the least expensive blood‐conservation practices because it is a ‘point‐of‐care’ strategy that is performed in the operating room at the time of surgery. As ANH and preoperative autologous donation (PAD) have equivalent efficacy but ANH is much less expensive than PAD, ANH is the logical choice for the procurement of autologous blood.

A nondamaging blood sampling technique for waterfowl embryos

Development of minimally invasive techniques to collect blood in free-living birds is desirable for both ethical and conservation reasons. We describe a new, simple technique for collecting blood samples from waterfowl eggs that involves recovering blood from a small hemorrhage after puncturing the shell of eggs showing the first signs of hatching. This technique allowed us to obtain a large number of blood samples from hatchling Greater Snow Geese (Chen caerulescens atlantica) and did not require the sacrifice or retention of any individuals, thus minimizing stress. High-quality blood samples for genetic studies were obtained, with no adverse effects on either embryos or the post-hatch survival of young geese. SINOPSIS. Una nueva t´ ecnica para obtener muestras de sangre de embriones de gansos Esy deseable, en aras de conservar, el desarrollar tpoco invasivas para tomar muestras de sangre de aves vivas. Describimos una tpara tomar muestras de sangre de huevos de gansos que consiste en tomar sangre de la pequehemorragia producida por una puncidel cascaron cuando se observan lo primeros signos de eclosionar. Este tpermiti´ o tomar una muestra considerable de sangre de huevos de ganso (Chen caerulescens atlantica) sin requerir sacrificar o retener a ning´ un individuo, minimizando de esta forma el estrcausando por lo ´ ultimo. Las muestras tomadas fueron de alta calidad para angensin que se observara efecto adverso alguno en los embriones o la supervivencia de los gansitos producto de los huevos manipulados y que eclosionaron.

Improving the bacteriological safety of platelet transfusions

Despite the increased application of aseptic techniques for blood collection and the preparation of platelet concentrates, morbidity and mortality arising from the transfusion of bacterially contaminated allogeneic platelet products persist. This problem exists because stored platelet concentrates represent a nearly ideal growth medium for bacteria and because they are stored at temperatures (22° ± 2°C) that facilitate bacterial growth. The presence of bacteria in blood components including platelets has been a problem for many decades and currently is the most common microbiological cause of transfusion-associated morbidity and mortality. A variety of strategies have been devised and/or proposed in an attempt to try to reduce the risk of transfusion-associated sepsis. These include pretransfusion bacterial detection, efforts to reduce the likelihood of bacterial contamination, the optimization of blood product processing and storage, reducing recipient exposure, and the introduction of pathogen inactivation methodology. With regard to doing bacterial detection, a number of automated detection systems have become available to test for contaminated platelet components, but their utility to some extent is restricted by the time they take to indicate the presence of bacteria and/or their lack of sensitivity to detect initially low bacterial loads. A variety of other approaches has been shown to reduce the risk of bacterial contamination and include filtration to remove leukocytes and bacteria, diversion of the initial aliquot of blood during donation, and improved donor skin disinfection. Platelet pathogen inactivation methods under investigation include the addition of L-carnitine, γ-irradiation, riboflavin plus UVA irradiation, and amotosalen HCl plus UVA irradiation. The latter process is licensed for clinical use with platelets in some countries in Europe. All of these approaches, either collectively or individually, hold considerable promise that the prevalence of adverse events associated with bacteria in platelet products will decline significantly in the very foreseeable future.

Specificity of the neuroendocrine response to orgasm during sexual arousal in men.

We have demonstrated that sexual activity produces transient sympathoadrenal activation and a pronounced, long-lasting increase in prolactin in men and women. However, by analyzing endocrine alterations at 10-min intervals, a precise assignment of these changes to the pre-, peri- and postorgasmic periods was not possible. Thus, the current study aimed to accurately differentiate the endocrine response to sexual arousal and orgasm in men using an automatic blood collection technique with 2-min sampling intervals. Blood was drawn continuously before, during and after orgasm over a total period of 40 min in 10 healthy subjects and were compared with samples obtained under a control condition. Sexual activity induced transient increases of plasma epinephrine and norepinephrine levels during orgasm with a rapid decline thereafter. In contrast, prolactin levels increased immediately after orgasm and remained elevated throughout the experiment. Although oxytocin was acutely increased after orgasm, these changes were not consistent and did not reach statistical significance. Vasopressin, LH, FSH and testosterone plasma concentrations remained unaltered during sexual arousal and orgasm. These data confirm that prolactin is secreted after orgasm and, compared with oxytocin, seems to represent a more reliable and sustained marker for orgasm in man. The results further reinforce a role for prolactin either as a neuroendocrine reproductive reflex or as a feedback mechanism modulating dopaminergic systems in the central nervous system that are responsible for appetitive behavior.

Differences in Umbilical Cord Blood Units Collected during Cesarean Section, before or after the Delivery of the Placenta

Umbilical cord blood is largely employed as an alternative source of stem cells in the treatment of hemato-oncological diseases. Current results show that the success rate of purified umbilical cord blood engraftment is comparable to that obtained using bone marrow, and it is directly related to the number of pluripotent stem cells transplanted. The technique of fetal blood collection varies among different umbilical cord blood banks. Many authors collect umbilical cord blood during vaginal delivery, after placental detachment, while others collect it while the placenta is still within the uterus. In a previous randomized trial, we showed a greater collection of umbilical cord blood before placental detachment during vaginal delivery. The present study was performed to determine whether umbilical cord blood collection before placental detachment (group A) during cesarean section is superior to that after placental delivery (group B) puncturing the umbilical vein once and using a closed bag system. To accomplish this, 47 pregnant women subjected to cesarean section were enrolled in the study. Twenty-one of them were allocated to group A, while the remaining 26 formed group B. The volume of umbilical cord blood collected from the patients of group A was greater than that collected from patients of group B. The cord blood volume collected was 90.7 ± 6.0 versus 60.9 ± 13.7 ml; the cord blood nucleated cell number was 10.1 ± 1.2 × 10⁸ vs. 7.1 ± 0.8 × 10⁸; and the mean cord blood CD34+ cell number was 20.0 ± 6.0 × 10⁵ vs. 16.4 ± 2.4 × 10⁵, respectively.

Ex vivo induction of cytokine mRNA expression in human blood samples

The interest in the quantitative analysis of cytokine mRNA profiles has increased substantially in recent years. This is based on the potential use of basal cytokine mRNA expression as sensitive markers for in vivo lymphocyte activation in a variety of clinical settings. However, it is less well known to what extent differences in blood collection and preparation techniques may cause ex vivo alteration of quantitative cytokine mRNA levels. We therefore evaluated the effect of blood sampling and the impact of cell separation on interleukin (IL)-2, IL-4, interferon (IFN)-γ and tumour necrosis factor (TNF)-α mRNA expression in an intraindividual study design ( n=8). Two different blood sampling procedures were applied. A whole blood sample 1 was collected by constant moderate blood flow into a blood collection tube containing lithium–heparin. Moreover, a second sample from the same donor was collected by a 5-fold acceleration of blood flow. Furthermore, peripheral blood mononuclear cell (PBMC) were isolated from the first whole blood sample by density separation over Ficoll-Hypaque. The quantification of cytokine mRNA expression was performed by real-time PCR in native whole blood/PBMC samples or unstimulated cultures. We found a significant increase of IL-2, IL-4 and TNF-α mRNA expression ( P=0.018, P=0.028, P=0.018) in whole blood samples collected by rapid sampling. The isolation of PBMC by density gradient separation prompted on upregulation of the mRNA levels of IL-2, IL-4 and TNF-α 5–9-fold ( P=0.018, P=0.018, P=0.018). In contrast, IFN-γ mRNA expression was not significantly influenced by differences in blood sample preparation. Our data clearly demonstrate that differences in the blood sampling technique or cell separation should be considered as important factors for non-physiological ex vivo induction of cytokine mRNA expression. The current data emphasize the need for data on the impact of ex vivo variation in order to extract reliable and consistent information, particularly when cytokine mRNA expression data from healthy blood donors are included in clinical studies.

Blood conservation in patients undergoing non-cardiac surgery

Concerns about the safety, inventory, and cost of allogeneic blood have led to a renewed interest in blood conservation. Autologous blood collection techniques, including preoperative autologous donation, acute normovolemic hemodilution, and perioperative blood recovery are routinely used as alternatives to allogeneic transfusion. In the future, these techniques may be combined with pharmacological strategies, such as presurgical erythropoietin therapy or red cell substitutes, to reduce further the need for allogeneic blood.

Measurement of blood serum cyclosporine levels using capillary "fingerstick" sampling: a validation study

Abstract Capillary blood sampling as a means of monitoring blood cyclosporine levels has replaced venipuncture in some medical centers. As the validity of capillary venipuncture for analysis of cyclosporine has not been documented, we sought to validate the capillary blood collection technique by comparing it with serum samples collected simultaneously by venous phlebotomy. Forty paired capillary-and venous samples were collected from 36 cardiac transplantation patients and analyzed, using a polyclonal immunoassay. The values obtained were compared using regression correlation. The correlation coefficient for all 40 samples was 0.859. However, we discovered that the first 7 capillary specimens were processed incorrectly. The correlation coefficient for the other 33 samples was 0.995 (99 % confidence interval 0.987-0.998). The excellent correlation between serum samples obtained from capillary sampling and from venous sampling, together with the ease of obtaining capillary blood specimens, make “fingerstick” sampling the method of choice for monitoring cyclosporine levels in infants and children.

Sample Collection and Clinical Pathology of Reptiles

Clinical pathology techniques can guide the clinician to diagnose and treat the reptile patient successfully. Text, tables, pictures, and diagrams in this article describe blood and cytology collection techniques and interpretation for the common pet reptile species.

Ethylenediaminetetraacetic Acid-Dependent Pseudothrombocytopenia in a Cardiac Surgical Patient

Ethylenediaminetetraacetic Acid-Dependent Pseudothrombocytopenia in a Cardiac Surgical Patient

Evidence that the mediobasal hypothalamus is the primary site of action of estradiol in inducing the preovulatory gonadotropin releasing hormone surge in the ewe.

Although a neural site of action for estradiol in inducing a LH surge via a surge of GnRH is now well established in sheep, the precise target(s) for estrogen within the brain is unknown. To address this issue, two experiments were conducted during the breeding season using an artificial model of the follicular phase. In the first experiment, bilateral 17beta-estradiol microimplants were positioned in either the medial preoptic area (MPOA) or the mediobasal hypothalamus (MBH), and LH secretion was monitored. An initial negative feedback inhibition of LH secretion was observed in ewes that had estradiol microimplants located in the MPOA (6 of 6 ewes) or caudal MBH in the vicinity of the arcuate nucleus (4 of 4). In contrast, a normal LH surge was only found in animals bearing estradiol microimplants in the MBH (5 of 10). Detailed analysis of estradiol microimplant location with respect to the estrogen receptor-alpha-immunoreactive cells of the hypothalamus revealed that 4 of the 5 ewes exhibiting a LH surge had microimplants located bilaterally within or adjacent to the area of estrogen receptor-expressing cells of the ventromedial nucleus. Two of these ewes exhibited a LH surge without showing any form of estrogen negative feedback. In the second experiment, we used the technique of hypophyseal portal blood collection to monitor GnRH secretion directly at the time of the LH surge induced by estradiol delivered either centrally or peripherally. Central estradiol implants induced the GnRH surge. The duration and mean plasma concentration of GnRH during the surge were not different between animals given peripheral or central MBH estradiol implants. Cholesterol-filled MBH microimplants did not evoke a GnRH surge. We conclude that the ventromedial nucleus is the primary site of action for estradiol in stimulating the preovulatory GnRH surge of the ewe, whereas the MPOA and possibly the caudal MBH are sites at which estrogen can act to inhibit LH secretion. These data provide evidence for the sites within the ovine hypothalamus responsible for mediating the bimodal influence of estradiol on GnRH secretion and suggest that different, and possibly independent, neuronal cell populations are responsible for the negative and positive feedback actions of estradiol.

Blood transfusions in the dog and cat Part 1. Blood collection techniques

THE use of blood transfusions in veterinary practice has increased dramatically in recent years. Practitioners therefore need to be able to collect and administer blood safely when a transfusion is...

THE EFFECTS OF ANTICOAGULATION AND PROCESSING ON ASSAYS OF IL-6, sIL-6R, sIL-2R AND SOLUBLE TRANSFERRIN RECEPTOR

Levels of plasma cytokines, their receptors or immune-related peptides, are used in experimental and clinical medicine. However, no standards are available regarding the use of anticoagulants or blood processing including the technique of blood collection or time delay between blood sampling and centrifugation.Blood was collected from 10 patients in order to assay interleukin 6, soluble interleukin 6 receptor, soluble interleukin 2 receptor and soluble transferrin receptor by enzyme-linked immunosorbent assay (ELISA). At the time of sampling, five different collecting tubes were used: allowing coagulation, in the presence of EDTA, with EDTA being administered to the serum after its separation, in citrated and in heparinized blood. Blood was centrifuged 10 min after collection and separated serum or plasma was immediately frozen at −20°C. To study the effect of time delay between blood sampling and processing, blood from 11 other patients was sampled, allowing coagulation; or in EDTA tubes and stored at 4°C. The blood was centrifuged 10 min, 8 h and 24 h later, and the separated serum or EDTA plasma was stored at −20°C until thawed for protein determination.Plasma interleukin 6 and soluble interleukin 6 receptor concentrations did not depend on the type of anticoagulant used or the time delay between sampling and processing. Soluble interleukin 2 receptor concentrations were not influenced by time delay before centrifugation, but concentrations were increased 10-fold in EDTA plasma exclusively when EDTA had contact with blood cells. Soluble transferrin receptor concentrations rose progressively with storage time before centrifugation. Contact of EDTA to whole blood or serum resulted in the same increase of soluble transferrin receptor concentration.The results suggest that standardization of the use of anticoagulant and time delay before centrifugation and serum or plasma separation is necessary for soluble interleukin 2 receptor and soluble transferrin receptor.

Capillary blood collection for neonatal screening tests should require a certificate of competence

Fourteen years ago, Patricia Townshend (1980) suggested that, “blood letting by heel prick should become a procedure that requires a certificate of competence”. To date, her suggestion has not been adopted, as competency is not a requirement to undertake this procedure. However, over the years many institutions have introduced accreditation for similar skills, such as diabetic monitoring and venepuncture. As a consequence, I believe standards of patient care have improved immensely. No longer can we assume all midwives are using the same techniques for neonatal capillary blood collection. From my observations of varied practice, such as twisting the blade and making multiple punctures for the one screening test, coupled with the poor quality of blood spot specimens frequently received by the central screening laboratory, it could be suggested that the introduction of accrediation for this skill, is well justified.

Hypophyseal Portal Blood Collection for Analysis of GnRH Neurosecretion in Shiba Goats.

Sequential sampling of hypophyseal portal blood was performed in the Shiba goat. The technique for portal blood collection originally developed for sheep was adapted with some modification. Portal blood was withdrawn by lesioning a part of the pituitary portal vessels via a collection apparatus placed beforehand at the rostral surface of the anterior pituitary gland. In four ovariectomized goats, portal blood was collected successively at 5 min intervals for several hours to examine the neurosecretory dynamics of hypothalamic GnRH. A pulsatile pattern of GnRH discharge into the pituitary portal circulation was clearly observed. Mean ± SD inter-pulse intervals in each of the 4 animals were 29.0 ± 2.0 min, 38.8 ± 2.2 min, 29.3 ± 1.7 min and 23.9 ± 2.1 min. Each GnRH pulse was associated with an LH pulse, and temporal correlation between the GnRH and LH pulses was consistent despite differing pulse frequencies among individual animals. Thus the technique for continuous pituitary portal blood collection from conscious goat has become available.

Neuroendocrine and metabolic adaptation of two cattle breeds to the physical stress of climbing to mountain pastures

One of the most important stress factors in the semi-extensive systems of cattle husbandry in the Pyrenees is when animals walk up to the pasture in the spring. At this time, several diseases, such as degenerative myopathies, may appear, and breed differences in sensitivity to pathological conditions have been observed. The aim of our experiment was to study the neuroendocrine and metabolic responses of Pirenaica and Parda Alpina breeds when they were submitted to the physical stress of climbing to mountain pastures after spending the winter housed indoors without exercise. Both rustic breeds were grown under exactly the same nutritional and handling conditions for meat production. Blood samples of 32 cows of each breed and their calves were obtained the day before walking up to the pasture (A), just when they arrived (B) and the day after (C). Animals were adapted to the blood collection technique, which was always carried out around 12:00 h. Cows and calves travelled by foot 14 km, climbing a vertical height of 600 m in approximately 3 h. Plasma cortisol values in Pirenaica cows were significantly higher (P < 0.001) than in Parda Alpina cows under basal conditions (A: 23.8 f 13.1 vs. 12.1 + 5.9). After exercise there were no significant differences (B: 14.1 rf: 7.4 vs. 13.4 f 8.0; C: 9.1 + 4.7 vs. 8.4 f 3.6). As animals were habituated to blood collection, probably Pirenaica cows have a true high baseline cortisol concentration, which is not attributable to a more intensive reaction to the act of being restrained and subjected to venipuncture. This fact might be related to the nervous character of the Pirenaica breed. However, these differences were not found in both breeds of calves. In young animals the more important differences were observed in muscle enzyme activities (creatine phosphokinase, lactate dehydrogenase, aspartate aminotransferase) which were always higher in Pirenaica calves in basal conditions and after exercise. This might explain why Pirenaica calves often present cardiomuscular problems when they walk up to the pasture. In cows, there were also significant differences in haematological values between both breeds, these values being parameters always higher in Pirenaica cows. Our results show that the Pirenaica breed displays in general higher neuroendocrine and metabolic indices of stress, but further studies must be done in order to define more precisely the psychobiological profile of both breeds and improve the adaptation of animals to extensive conditions.

Two capillary blood-collection techniques for estimating glycohemoglobin compared

Journal Article Two capillary blood-collection techniques for estimating glycohemoglobin compared Get access S K Baglin, S K Baglin Search for other works by this author on: Oxford Academic Google Scholar A S Brown A S Brown Search for other works by this author on: Oxford Academic Google Scholar Clinical Chemistry, Volume 41, Issue 2, 1 February 1995, Pages 330–332, https://doi.org/10.1093/clinchem/41.2.330 Published: 01 February 1995

L'érythroaphérèse préopératoire permet-elle d'éviter la transfusion homologue ?

The risk of transfusion-related viral infections promotes techniques intended to reduce such a risk. Preoperative erythroapheresis is an autologous blood collection technique which minimizes the inconvenience of repeated predeposit autologous donations. Erythroapheresis has been performed in 451 patients, 24 to 72 hours before surgery using a discontinuous flow cell separator (PCS+Haemonetics). The mean collected red cells volume was 485 ml (250-900) and was replaced by hydroxy-ethyl-starch (ELOHES). Homologous blood transfusion was avoided in 431 patients (95,6%). This technique is indicated for surgery with an expected blood loss between 1000 and 1500 ml or when the patient cannot be included in a preoperative blood donation programme. Preoperative erythrocytapheresis is a simple and well tolerated procedure. To have recourse to apheresis techniques allows to increase the number of autologous transfused patients.

Patterns of release of oxytocin, prolactin, insulin and LH in lactating sows, studied using continuous blood collection technique.

To describe the patterns of release of oxytocin, prolactin, insulin and LH in relation to suckling and to investigate relationships between them, blood was collected continuously from four sows at a rate of 1 ml per min for 12 h (0900-2100 h) on day 10 (D10) and day 20 (D20) of lactation. The 12-h period was divided into four smaller periods: P1 (0900-1200 h), P2 (1200-1500 h), P3 (1500-1800 h) and P4 (1800-2100 h). The number of nursings occurring during a 12-h period tended to decrease between D10 (17.3 +/- 0.3) and D20 (16.0 +/- 0.3). About 40-50% of all sucklings induced a significant rise in plasma oxytocin, while prolactin gradually increased following each suckling. Decrease in the levels of oxytocin (14.1 +/- 0.4 vs 7.1 +/- 0.4 pmol/l, P < 0.001), prolactin (12.9 +/- 0.3 vs 7.9 +/- 0.1 micrograms/l, P < 0.001) and insulin (24.8 +/- 1.7 vs 14.4 +/- 1.7 mU/l, P < 0.001) were observed between D10 and D20. Average and basal levels of LH and the number of LH pulses increased (P < 0.01) between D10 and D20. A negative correlation between number of sucklings and number of LH pulses was demonstrated on D20 (r = -0.59, P = 0.07). There was a positive correlation between levels of LH and oxytocin on D10 (r = 0.54, P = 0.001) and between prolactin and insulin levels on D20 (r = 0.34, P = 0.05).