| Summary: | The cardiac puncture technique for obtaining relatively large volume (50–150 µL) blood samples from sedated rodents has been used in research for nearly a century. Historically, its use to phlebotomize and then release live rodents was more common. However, recently its use in a non-terminal capacity frequently imparts negative connotations in part because exsanguination of sedated animals via cardiac puncture is now an American Veterinary Medical Association-approved euthanasia technique. This association has resulted in ethical concerns by manuscript reviewers and in a few instances, outright refusal by some peer-reviewed journals to publish research that utilized the technique. To counter the perceived negative associations with its non-terminal use, we summarized nearly two decades (2001–2019) of capture and handling data throughout Connecticut, resulting in over 7000 cardiac punctures performed on nearly 5000 sedated, live-captured and released <i>Peromyscus</i> spp. We show that our total handling mortality rate (3.7%) was comparable, if not lower, than similar field studies that utilized other phlebotomy techniques. Many public health, integrated tick management, and vector-borne disease ecology studies require samples from individual wild-caught <i>Peromyscus</i> spp. over time to determine intervention efficacy and pathogen infection monitoring, and in such field studies, post-operative care is not an option. Proper execution of cardiac puncture does not increase susceptibility of individuals to predation upon release as can potential ocular abnormalities or infections that can occur as the result of use of other techniques. We posit that neither exsanguination nor resulting euthanasia are requirements of cardiac puncture and that its use is entirely appropriate for obtaining blood samples from live-captured and released <i>Peromyscus</i> spp. Properly performed cardiac puncture is an excellent technique to obtain blood samples from sedated, individual <i>Peromyscus</i> spp. on multiple appropriately-spaced occasions over single trapping seasons while keeping animal welfare a top priority.
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