Evaluation of 3 Alcohol-based Agents for Presurgical Skin Preparation in Mice.

Appropriate aseptic technique is a crucial component of rodent survival surgery. Ease of technique, surgical space constraint, batch surgery, and cost are factors that may affect researcher compliance with appropriate aseptic technique. The first part of this study compared 3 antiseptic preparation agents with the standard triplicate application of povidone-iodine and alcohol. Euthanized mice (n = 40) were shaved on the dorsum, and culture swabs were taken for RODAC plating and bacterial identification. Shaved sites were prepared by using one of the 4 antiseptic preparation agents. Culture samples were obtained immediately and at 20 min after antiseptic preparation. In the 2nd part of the study, 8 mice (n = 2 per group) were prepared for a survival surgical procedure by using one of the 4 antiseptic preparation agents to evaluate whether the antiseptic preparation agents caused skin irritation or impaired healing. Results from this study indicated that all 3 of the antiseptic agents evaluated were equally effective at reducing bacterial populations immediately and at 20 min after preparation. Histopathologic examination of the incision sites revealed signs of normal healing without lesions adjacent to the incision site. We conclude that all 3 of the products evaluated are comparable to traditional povidone-iodine and alcohol as agents for aseptic preparation of surgical sites.

Bias due to Preanalytical Dilution of Rodent Serum for Biochemical Analysis on the Siemens Dimension Xpand Plus.

Clinical pathology testing of rodents is often challenging due to insufficient sample volume. One solution in clinical veterinary and exploratory research environments is dilution of samples prior to analysis. However, published information on the impact of preanalytical sample dilution on rodent biochemical data is incomplete. The objective of this study was to evaluate the effects of preanalytical sample dilution on biochemical analysis of mouse and rat serum samples utilizing the Siemens Dimension Xpand Plus. Rats were obtained from end of study research projects. Mice were obtained from sentinel testing programs. For both, whole blood was collected via terminal cardiocentesis into empty tubes and serum was harvested. Biochemical parameters were measured on fresh and thawed frozen samples run straight and at dilution factors 2-10. Dilutions were performed manually, utilizing either ultrapure water or enzyme diluent per manufacturer recommendations. All diluted samples were generated directly from the undiluted sample. Preanalytical dilution caused clinically unacceptable bias in most analytes at dilution factors four and above. Dilution-induced bias in total calcium, creatinine, total bilirubin, and uric acid was considered unacceptable with any degree of dilution, based on the more conservative of two definitions of acceptability. Dilution often caused electrolyte values to fall below assay range precluding evaluation of bias. Dilution-induced bias occurred in most biochemical parameters to varying degrees and may render dilution unacceptable in the exploratory research and clinical veterinary environments. Additionally, differences between results obtained at different dilution factors may confound statistical comparisons in research settings. Comparison of data obtained at a single dilution factor is highly recommended.

Alterations due to dilution and anticoagulant effects in hematologic analysis of rodent blood samples on the Sysmex XT-2000iV.

BACKGROUND: Clinical pathology of rodents is hindered by sample volume limitations. A single whole heparinized blood sample is often submitted for hematologic and clinical chemistry analysis in exploratory research settings, and sample dilution may be required. Published information on the potential impact of sample dilution and heparin use on hematology variables in rodents is sparse. OBJECTIVES: The purpose of the study was to evaluate the effects of sample dilution and of anticoagulant on hematologic analysis of mouse and rat blood samples on the Sysmex XT-2000iV. METHODS: Mice and rats were obtained from various ends of study research projects, and whole blood was collected via terminal cardiocentesis in lithium heparin, and additionally in EDTA when paired samples were obtained from rats. Hematology analytes were measured on the Sysmex XT-2000iV straight and diluted from ×2 to ×5. RESULTS: Significant differences between heparinized samples analyzed straight vs diluted were found for MCV and MCHC, with a bias for several additional variables. Significant differences between paired heparinized and EDTA-anticoagulated samples at each dilution point were found for most variables, with the largest differences found in platelet count. Evidence of platelet clumping presumably due to heparin exposure was noted in numerous samples. CONCLUSIONS: Dilution-induced changes occur in some hematologic variables and may render dilution unacceptable in the exploratory research environment. Many variables, most notably platelet count, differ based on the anticoagulant used, and values from heparinized vs EDTA-anticoagulated samples should not be directly compared.