Ultrasound-guided blood sampling of rabbit fetuses.

A rabbit animal model for hemolytic disease of the newborn has been previously described. However, evaluating the effects of this disease was limited to histologic and hematologic examinations of liveborn kitlings. To assess the feasibility of in utero blood sampling, we performed ultrasound-guided cardiac sampling of 50 fetuses in 16 New Zealand White does on days 26 and 27 of gestation. The overall rate of successful sampling was 80%. The procedure-related mortality declined to 35% by the third phase of the study. The mean (+/- SD) hematocrit (%) and reticulocyte values (#/100 RBCs) on day 26 were 26.3 +/- 3.3 and 35.6 +/- 5.1, respectively; values on day 27 were 31.3 +/- 4.9 and 27.5 +/- 7.6. The results of this study suggest that hematologic data can be obtained from rabbit fetuses in the majority of cases with only moderate fetal loss.     

Lateral flow devices for samples collected by straw sampling method for postmortem canine rabies diagnosis

The direct fluorescent antibody test (dFAT) using brain sample after opening the skull is the standard rabies diagnostic test in animal rabies. However, it is not feasible in many resource-limited settings. Lateral flow devices (LFD) combined with a simple sampling methodology is quicker, simpler, and less hazardous than the standard test and can be a useful tool. We conducted a prospective on-site study to evaluate the diagnostic accuracy of the LFD with the straw sampling method compared with that of the dFAT with the skull opening procedure for post-mortem canine rabies diagnosis. We collected 97 rabies-suspected animals between December 1, 2020 and March 31, 2021. Among the 97 samples, 53 and 50 cases were positive tests for dFAT and LFD, respectively. The sensitivity and specificity of LFD with straw sampling method were 94.3% (95% confidence interval [CI], 84.3–98.8%) and 100% (95% CI, 92.0–100%), respectively. The performance of LFD by the straw sampling method showed relatively high sensitivity and 100% specificity compared with that of dFAT performed on samples collected after opening the skull. This methodology can be beneficial and is a strong tool to overcome limited animal surveillance in remote areas. However, because of our limited sample size, more data using fresh samples on-site and the optimizations are urgently needed for the further implementation in endemic areas.     

Use of arterialized venous blood sampling during incremental exercise tests.

Close agreement between arterialized venous and arterial pH, PCO2, and lactate has previously been demonstrated during steady-state exercise. The purpose of the present study was to compare arterialized venous and arterial pH, PCO2, K+, lactate, pyruvate, and epinephrine during the constantly changing circumstances of an incremental exercise test. Eight normal subjects undertook an incremental exercise test (increasing by 20 W/min) to exhaustion on a cycle ergometer during which simultaneous arterial and arterialized venous samples were drawn over the last 20 s of each work load. Linear regression of arterialized venous on arterial values showed that r varied from 0.97 to 0.99 for the variables examined and, therefore, showed that accurate estimates of arterial values could be made from the arterialized venous results during incremental testing. For many purposes it could be assumed that arterialized venous values equaled arterial values without serious error.     

Simultaneous blood and biliary sampling of esculetin by microdialysis in the rat

Biliary excretion and intestinal reabsorption in enterohepatic circulation play major dispositional roles for some drugs. To circumvent multiple blood sampling and interruption of enterohepatic circulation in conventional biliary cannulation, the present study utilized the minimally invasive sampling technique of microdialysis in pharmacokinetics and biliary excretion studies. Microdialysis probes were inserted into the jugular vein and bile duct in the anesthetized rat for simultaneous and continuous sampling following intravenous administration of esculetin, a bioactive coumarin derivative. Placements of the microdialysis probes were designed to minimize obstruction to normal flows of the body fluids. Separation and quantitation of esculetin in the dialysates were achieved using high performance liquid chromatography (HPLC) coupled to UV detection. The results indicated higher drug concentrations in the bile than in the blood, suggesting active biliary excretion. The study also provided an example of successful application of in vivo microdialysis as an interesting and feasible alternative for pharmacokinetics and biliary drug excretion studies.     

A refined method for sequential blood sampling by tail incision in rats

Levels of endogenous or administered substances can be estimated by blood sampling. This allows an evaluation of the relationship between clinical signs, physiological parameters, pharmacological treatments and behaviour of the animal. We show that blood samples can be taken occasionally as well as sequentially by means of a small incision at the end of the rats' tails. Up to 300 microl of blood can be collected within 90 s. The advantages of this method are: (i) anaesthesia and surgery or restraint of the animal are not necessary; (ii) the procedure can be considered stress-free as indicated by the low, basal levels of the stress hormone corticosterone, even with frequent sequential blood sampling over 3 h; and (iii) it can be used for longitudinal studies allowing intra-individual comparisons over months and even years. Blood samples collected via an intravenous catheter and, at the same time, by our tail incision method resulted in comparable amounts of corticosterone. Moreover, we consider the tail incision method for rats to be 'animal-friendly' and a real alternative to other conventionally used blood sampling techniques.     

Home blood sampling for plasma glucose assay in control of diabetes.

Estimation of plasma glucose in home blood samples is needed to improve diabetic control. Sufficiently precise measurements on capillary blood were obtained by (a) storing Reflotest glucose-oxidase strips in a desiccant container before reading and (b) collecting blood samples into a simple vacuum bottle containing potassium fluoride (assay of sodium content indicating volume of plasma collected). The precision of the methods (+/- 1 SD) was +/-0.35 mmol/1 (+/-6.3 mg/100 ml). Clinical reliability was assessed by measuring the basal plasma glucose concentration at home on different mornings in patients with maturity-onset diabetes, the day-to-day variation (+/- 1 SD) being +/-0.73 and +/-0.92 mmol/1 (+/-13.2 and +/-16.6 mg/100 ml) respectively. The mean basal plasma glucose concentration in all 84 patients with maturity-onset diabetes from three general practices was 8 mmol/1 (144 mg/100 ml), 44 of the values exceeding 6 mmol/1 (108 mg/100 ml). Improving control by monitoring the basal plasma glucose concentration in maturity-onset diabetes might help to prevent diabetic complications.     

A blood sampling technique for prehatched chicks

ABSTRACT Collecting blood from neonatal or prehatched chicks is sometimes necessary, requiring specialized blood collection techniques and skills. However, such sampling can be difficult and could potentially have adverse effects. I developed a method for collecting blood from chicks still in an egg. The technique involves enlarging the pipping hole, removing the chick's head to take blood from a jugular vein, and returning the chick into its original position in the egg to complete hatching back in the nest. I used this technique on Crested Tern (Thalasseus bergii) chicks (N= 13), with no apparent adverse effects to growth or short‐term survival. The amount of blood extracted (0.1–0.15 ml) was sufficient for a blood smear, hematocrit, and total white blood cell count. This method should be useful for eggs as small as 40 mm in length; eggs smaller than this would likely not be good candidates for this technique because eggshells may be too thin to safely handle. This technique may be useful for investigators studying the immunological and endocrinological transition between embryos and hatchlings.     

Long-term blood sampling technique in the marmoset

A simple and reliable method for long-term blood sampling from the restrained marmoset is described. Cannulae were chronically inserted into the internal jugular vein, exteriorized and housed in a backpack system. The cannulae remained patent for 1-7 months. This system allows for rapid collection of blood samples and hence detailed analysis of circulating hormone levels.     

The effect of sampling technique on some blood parameters in the rat.

When used to restrain rats while obtaining cardiac blood, there was little difference between the effects of ether, pentobarbitone sodium, and fentanyl plus droperidol on acid-base balance, haemoglobin, haematocrit, plasma protein, calcium and magnesium. The use of manual restraint increased blood acidity, plasma protein, calcium and magnesium, but decreased blood glucose. Haemoglobin and haematocrit values were higher for tail blood than for blood removed from the heart or abdominal aorta.     

A technique for repeated sampling of the blood of individual resting fish.

A dorsal cannulation technique is described. It has been employed for repeated blood sampling in unanaesthetized rainbow trout (Salmo gairdneri) kept singly in special receptacles described in the paper. The level of the studied haematological parameters (Haematocrit, Hb, glucose, lactate, K+, Na+, Ca2+) differed between fish kept in receptacles for 1 week and free-swimming fish, most probably owing to differences in the motility of the fish. The receptacle seems to minimize visual and handling disturbances, and permits both the standardization of experimental conditions and quick and easy sampling via the dorsal aorta cannula. The general variation in the blood parameter values was very small compared with the previously reported variation in such values for rainbow trout.     

An improved cannulation technique for prolonged blood sampling of the American bullfrog.

A T-cannula was used to collect serial blood samples from the sciatic artery of the American bullfrog (Rana catesbeiana). The horizontal arm of the cannula was implanted in the artery and the vertical arm employed to collect blood samples. The T-cannula allowed blood flow to continue in the cannulated artery and provided long-term, repetitive blood samples from conscious, caged, but otherwise unrestrained and undisturbed animals. The cannula functioned for at least 14 days, the maximum period investigated. Athough developed for studies on amphibians and reptiles, the technique appeared to be applicable, with appropriate modifications, to a variety of small animals.     

Thyrotrophin-releasing hormone inactivation by human postmortem brain

The mechanisms of inactivation of thyrotrophin-releasing hormone (TRH) by peptidases in several areas of normal human postmortem brain have been investigated by radioimmunoassay and high-performance liquid chromatography. Of the several brain regions studied, the cerebral cortex (Brodman's area, BA10) had the highest TRH-degrading activity in both subcellular fractions. Deamidated-TRH (TRH-OH) was the only product formed by the soluble fraction whereas the histidyl-proline diketopiperazine, cyclo(His-Pro), and a small amount of TRH-OH were formed by the particulate fraction. Several centrally acting TRH analogues showed varying degrees of resistance to degradation by the peptidases in the two fractions, the most stable analogue being RX77368 (pGlu-His-3,3'-dimethyl(ProNH2]. Areas of human postmortem brain appear to contain two of the enzymes capable of degrading TRH, a proline endopeptidase forming TRH-OH and a pyroglutamyl aminopeptidase forming cyclo(His-Pro). The use of the assay procedures in further studies on the inactivation of TRH by peptidases from brain areas of patients with neurological disorders may provide complementary information on the dynamics of TRH in these disorders. The stability of the centrally acting TRH analogues may prove useful in examining their therapeutic potential.