Various puncture routes, veins, arteries, heart, are used to take blood in animals. For anatomical reasons, differences in blood composition are expected among puncture sites. However, this issue has been rarely assessed and contrasted results have been reported: strong effects of puncture site versus a lack of effect. We captured free-ranging freshwater turtles from different locations to compare the mean concentrations of 12 blood parameters (metabolites, hormone, ions, and enzyme) among three puncture sites: (1) a lateral branch of the jugular vein, (2) a dorsal subcarapacial cervical plexus (sometimes incorrectly referred as the 'cervical sinus' in the literature), and (3) a caudal plexus site (sometimes incorrectly referred as the 'caudal sinus'). Because we used very small syringes (27-30G), we were able to separate lymph, blood, or blood-lymph mixtures. Our results show very strong effects of puncture site and of mixture level (mean maximal difference between sites was 250 %). We also found strong sex and geographical effects. Typically, there were differences in concentrations of blood solutes sampled from the lateral jugular vein and subcarapacial plexus, mainly due to sampling a mixture of blood and lymph from the 'blood' at the subcarapacial site and pure blood from the lateral jugular site, and likewise, samples from the caudal site were highly variable due to often sampling a mixture of blood and lymph. These results have technical and fundamental implications, especially when performing comparative analyses. Further, by selecting precise puncture sites, physiological differences between lymph and blood compartments could be investigated.
Keywords: Corticosterone; Hemodilution; Lymph; Plasma metabolites; Turtle.