A simple technic for repeated collection of blood samples from mice.

A device for repeated collection of small blood samples from mice was constructed from a plastic syringe. Blood was collected into a 3.33 lambda capillary tube. Bleeding was stopped by a hemostat made from a rubber stopper. This technic allows easy collection of approximately 20 serial samples within an 8-hr period.     

A simple technique for blood collection in the opossum (Didelphis virginiana)

The left and right brachiocephalic veins were used for rapid collection of 5-10 ml blood samples in adult opossums. These vessels were also used to collect 0.5-1 ml blood samples in 120-day-old opossums. The animals were anaesthetized with 30 mg/kg of ketamine hydrochloride given intramuscularly prior to blood collection.     

A simple technique for electroelution of DNA from environmental samples.

A novel method was developed for the extraction of DNA from environmental material. Soil or sediment samples were encapsulated in agarose blocks and, following treatment with lysis reagents, the DNA was extracted by electroelution.     

An improved method for concentrating rotavirus from water samples

A modified adsorption-elution method for the concentration of seeded rotavirus from water samples was used to determine various factors which affected the virus recovery. An enzyme-linked immunosorbent assay was used to detect the rotavirus antigen after concentration. Of the various eluents compared, 0.05M glycine, pH 11.5 gave the highest rotavirus antigen recovery using negatively charged membrane filtration whereas 2.9% tryptose phosphate broth containing 6% glycine; pH 9.0 was found to give the greatest elution efficiency when a positively charged membrane was used. Reconcentration of water samples by a speedVac concentrator showed significantly higher rotavirus recovery than polyethylene glycol precipitation through both negatively and positively charged filters (p-value <0.001). In addition, speedVac concentration using negatively charged filtration resulted in greater rotavirus recovery than that using positively charged filtration (p-value = 0.004). Thirty eight environmental water samples were collected from river, domestic sewage, canals receiving raw sewage drains, and tap water collected in containers for domestic use, all from congested areas of Bangkok. In addition, several samples of commercial drinking water were analyzed. All samples were concentrated and examined for rotavirus antigen. Coliforms and fecal coliforms (0->1,800 MPN/100 ml) were observed but rotavirus was not detected in any sample. This study suggests that the speedVac reconcentration method gives the most efficient rotavirus recovery from water samples.     

A simple and efficient method for obtaining urine samples from rats

A method for collection of urine from rats was developed that is simple, reliable, and efficient. A 5 ml. polystyrene beaker is placed over the urethra and the base of the tail is stimulated with the fingers of one hand. Depending on the quality and quantity of urine needed the perineal area may be shaved and the beaker may be held by hand or attached with tape.     

A simple and inexpensive device for collecting urine samples from rats

Many studies require collection of metabolic wastes from laboratory animals, and oftentimes it is important that feces and urine be collected separately. The authors describe an easily assembled and inexpensive device that can be used to collect urine samples from rats without any invasive operations. The device affords reasonable separation of feces and urine.     

Unassisted blood collection from unanesthetized rats and gerbils

Blood collection is a common procedure in animal research. This column describes humane methods of collecting blood from the rat and the gerbil that can be carried out by a single person on an awake animal.     

A simple method for obtaining blood samples from mature frogs

A technique was developed for obtaining periodic blood samples from mature frogs. Up to 150 microliter of blood was collected at weekly intervals. Anesthetization was not required. The data compiled with this technique indicated that activity-induced hyperglycemia was minimal.     

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.     

Retro-orbital technique for blood collection from the ferret (Mustela putorius furo)

Blood samples were obtained from anesthetized ferrets of various ages, by venipuncture of the retro-orbital plexus with a heparinized capillary pipette. It is recommended that a cumulative collection of 20% of total blood volume not be exceeded for any two week period. This technique, when carefully performed on alternate orbits and at a frequency dictated by body weight and volume of blood needed, has been used repeatedly for months without apparent harm or distress to the animal.     

Rapid and simple screening of transgenic mice: novel extraction-free, filter-based PCR genotyping from blood samples

We evaluated the effectiveness of using Flinders Technology Associates (FTA) filter paper for the polymerase chain reaction (PCR) genotyping of transgenic mice. Tail prick blood sample dried on an FTA filter disc was processed for genomic PCR. It is easy and rapid to prepare DNA templates because the protocol is extraction-free and only requires minimal handling of wash briefly bloodstained FTA filter discs. Progeny of a transgene-positive founder mated with wild-type mice was screened for the presence of the transgene by the filter-based PCR using transgene-specific primers. The resulting amplicons with expected sizes of 3134 bp, 1152 bp, 877 bp and 688 bp were robust and reproducible, allowing a distinction between transgenic (n=44) and wild-type (n=47) mice showing no signal. The filter-based PCR screening took only half a day. The present study confirmed the validity and usefulness of the novel rapid extraction-free genotyping method.     

An improved and easy technique for polyamine determination in biological samples : Application to cell-free system from hypertrophied rat heart

An accurate, improved cation-exchange chromatographic method using o-phthalaldehyde and ultraviolet detection at 280 nm for the determination of free polyamines (putrescine, spermidine, spermine) has been developed. Different samples, such as the 105,000 g supernatant of reticulocyte or heart muscle, and KCl ribosomal wash containing initiation factors, can be analysed. The minor modification of reagents results in a good precision and sensitivity, which is demonstrated by a relative standard deviation of 5–9% and recoveries of 98%. This technique is of particular interest because it allows polyamine determination in biological samples with high concentrations of salt.     

A rapid method for repeated collection of blood from the tail vein of rats

A method was developed for obtaining serial blood samples from the tail vein of rats. Seventy microliters of blood were collected at the rate of approximately one sample per minute for the quantitative determination of whole blood clearance of radiolabeled compounds. The technique was rapid, reproducible, and kept stress to the test animals at a minimum.