Humane Use of Cardiac Puncture for Non-Terminal Phlebotomy of Wild-Caught and Released Peromyscus spp.

Simple Summary When researching tick-borne diseases and their management in the interest of improving public health, blood samples often need to be obtained from small rodents, which are the main source of the various pathogens that are picked up by ticks and can infect humans. In such research projects, animals are handled and released back into the environment with the least amount of harm done to ensure their continued survival. Post-sampling animal care is not an option on released animals as it is in a laboratory in a captive setting, therefore, sampling protocols need to reflect this fact. Blood sampling via cardiac puncture (sampling blood directly from the heart) tends to have a negative connotation because it is often associated with a procedure used for humane euthanasia in which sedated animals are bled to death per study protocols. We argue its use for obtaining blood samples is preferred in a field setting in which rodents are released. We show that our recapture and mortality rates rival or are better than other studies that utilize more familiar techniques. Death is not a requirement of its use and we suggest cardiac puncture for blood sampling is in the best interest of animal welfare because it does not make small rodents more prone to infection or negatively impact their vision or survival as can other blood sampling procedures. Abstract The cardiac puncture technique for obtaining relatively large volume (50-150 mu L) 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 Peromyscus 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 Peromyscus 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 Peromyscus spp. Properly performed cardiac puncture is an excellent technique to obtain blood samples from sedated, individual Peromyscus spp. on multiple appropriately-spaced occasions over single trapping seasons while keeping animal welfare a top priority.