The 2013 AVMA Guidelines for the Euthanasia of Animals recommends a chamber volume displacement rate of 10% to 30% per minute (v/min) when euthanizing small laboratory rodents with CO₂. Group euthanasia of mice is a common practice, and grouping strangers is often avoided to minimize distress; however, emotional contagion, which occurs between familiar animals but not strangers, has not been studied in the context of group CO₂ euthanasia. This study examined cagemate- and stranger-grouped mice exposed to 10%, 30%, or 50% v/min CO₂ to determine whether emotional contagion plays a role in this context and whether that role is influenced by CO₂ flow rate. Videos of adult male C57BL/6J mice exposed to different CO₂ flow rates were scored for durations of dyspnea, ataxia, and consciousness as well as the numbers of face pawing and jump behaviors. Blood was collected at time of unconsciousness and assayed for ACTH. Cagemates experienced significantly longer durations of conscious dyspnea and ataxia with 10% v/min CO₂ compared with 30% and 50% v/min. Similarly, strangers experienced significantly longer duration of conscious dyspnea with 10% v/min CO₂ compared with 30% and 50% v/min and significantly longer duration of ataxia with 10% compared with 50% v/min. Cagemates showed significantly more jumps with 10% v/min CO₂ compared with 30% and 50% v/min, whereas jumping was unaffected by CO₂ flow rate in strangers. We conclude that more potential for distress exists when cagemate and stranger mice are exposed to a 10% v/min CO₂ flow rate and that emotional contagion may contribute to distress in cagemates at this flow rate. Therefore, we propose that 30% v/min CO₂ should be used for euthanasia of mice, and that 50% v/min should also be considered humane.

Social housing of laboratory rodents is recommended whenever possible to encourage natural behavior and social dynamics. Several identification methods are used to distinguish rodents from one another. One of the most common means of identifying mice is ear punching. The effect of ear punching for identification or genotyping on the welfare of mice remains a concern, because this method negatively affects welfare in other species. To assess the influence of ear punching on the welfare of mice, we implanted telemetry units in 6 female Swiss–Webster mice and monitored heart rate, body temperature, and activity after various routine procedures. The physiologic and behavioral responses to restraint (by scruffing) only, restraint and ear punching, and routine handling for husbandry were evaluated. The mean heart rate of mice after receiving an ear punch was significantly higher than baseline values at 30 min after the procedure, and the mean body temperature was significantly increased over baseline for at least 1 h. The heart rate, body temperature, and activity levels of mice after scruffing only and routine handling did not differ from baseline values. The proportion of time mice spent head grooming, a potentially nocifensive behavior, was increased immediately after ear punching and began to decline by 60 min. We show that the physiologic stress of mice receiving an ear punch was greater than that from restraint (scruffing) alone, whereas behavioral indices of pain were unchanged, suggesting that ear punching causes a transient response in mice.

The methods and conditions for housing research mice have been the subject of many discussions and publications in recent years. At our institution, we began to explore these matters with the goal of reducing stress in the animals and yet maintaining an environment that more closely resembled their habitat in the wild and yet was acceptable to researchers and the technicians that cared for the animals. Through a series of small inhouse studies, we derived a method that allowed the animals to stay in their established environment for longer than the standard 1- or 2-wk period. After several empirical studies, we concluded that the mice could stay in the same cage for 16 wk or perhaps even longer. To achieve this outcome, we perfected a method of removing 75% of the existing cage bedding and replacing it with clean bedding every 2 wk. To substantiate the validity of the method, we conducted a major study that evaluated the conditions of the cage, cage environment and the animals for a 16-wk period. In the study, we compared all of these factors in the 16-wk cages to a set of cages that were completely replaced on a 2-wk cycle. The mice in our study appeared to experience decreased stress, and observation also revealed that the 16-wk method was associated with increased pup survival in several colonies. The revised 16-wk method appears to create mouse cage conditions that are no different than the current standard (that is, every 1 or 2 wk) methods of cage changing.

The Guide recommends sanitizing cage components, including microisolation cage tops (MCT), at a minimum of every 2 wk. Previously published data demonstrated that mouse MCT microbial loads do not increase until at least 2 wk and that sanitation can be delayed past 2 wk. How microbial loads differ on mouse compared with rat MCT, as well as across different ventilation systems, remains unclear. We hypothesized that MCT microbial loads would be higher in tops from rats compared with mice and would differ according to IVC ventilation system. We evaluated bacterial loads on MCT at serial time points to 90 d from static cages housing mice or rats and from rat and mouse cages on several ventilation systems (mice, 6; rats, 4). MCT were determined to have sufficiently elevated bacterial loads to necessitate changing based on either statistically significant changes in bacterial loads or values greater than 50 cfu. Across all ventilation systems, bacterial counts at 14 d were significantly higher on rat MCT compared with mouse MCT. Across the ventilation systems examined, rat MCT cfu remained similarly elevated from 14 d through 90 d. Mouse MCT total cfu were also stable across multiple ventilation systems yet remained lower than 50 cfu until at least 90 d. Patterns of bacterial species isolated from rat MCT were relatively consistent over time and ventilation system, whereas mice showed greater variability in both contexts. We found that 14 d is an appropriate sanitization time point for rat MCT, whereas the interval at which mouse MCT are cleaned can be extended to 90 d at least. Our data highlight interspecies differences in the accumulation of bacteria on MCT and that mouse MCT sanitation intervals for several housing systems can be extended beyond 14 d.

A stable environment is a prerequisite for animal research. The absence of a suitable laboratory rabbit environment at the gyrotron facility, with the nearest housing being 6.3 miles away, made it challenging to investigate ocular damage induced in rabbit eyes due to exposure to high-frequency millimeter waves. Because rabbits are prone to transportation stress, it was vital to keep them on-site during research. Here we describe the creation of the stable environment necessary to perform reliable and reproducible animal experiments, by using a cargo van parked at the gyrotron facility. To control the interior environment, we placed a window air conditioner, humidifier, dehumidifier, and photocatalyst deodorizer inside the cargo area without altering the original configuration of the vehicle. Rabbits were housed in individual cages for a maximum of 6 d. Microbial contamination in the air was evaluated by using a passive sampling method. The average numbers of bacterial and fungal colony forming units per dish were 0.2 and 4.7, respectively, indicating that the van was as clean as a nonbarrier animal facility. The average temperature was 20.5 ^°C (range, 17.8 to 22.6 ^°C), and the average relative humidity was 49.4% (range, 36.2% to 63.2%). The concentration of ammonia was consistently below the detection limit of 0.5 ppm. Other environmental conditions were within appropriate levels. Rabbits lost 6.4% ± 2.2% (n =52) of their initial body weight during the 13- to 14-h transport but recovered the lost weight within 48 h after arrival.

Neurophysiologic studies of NHP commonly involve their transfer from a housing enclosure to a laboratory by using a mobile chair. This transfer should be performed in a manner that is safe and minimizes stress for both animal and handler. The risk of harm associated with attempting to transfer these animals is increased when they are mature and naïve. We have modified previous chair designs and transfer methodologies to reduce this risk by maintaining a constant barrier between NHP and handler while providing control to the handler to facilitate chairing. Chair modifications were built inhouse, and a commercial, hydraulic lift table was used to dock the primate chair to home cages of different heights. The docking chair method was used with 8 adult, male rhesus macaques. A graduate student transferred the animals without complications. These modifications did not compromise existing features of the chair, they did not require training time in addition to that for the standard chairing method in our facility, and they improved safety. These refinements to a commonly used chair and transfer methodology support rapid habituation, safe transfer and reduced stress for both animal and handler. The refinements we describe mitigate the potential risk of harm during NHP transfers and thus advance animal welfare.

Federal regulations and policies require institutions to establish procedures for ongoing IACUC oversight of approved animal care and use program activities including animal procedures. To fulfill these requirements, research institutions implement postapproval monitoring (PAM) programs designed to assure compliance in animal activities. Although several references commenting on the requirement to conduct PAM are available, few publications discuss actual best practices for accomplishing PAM. Here we use information collected through a survey of large academic research institutions to identify common practices for conducting PAM reviews. Many similarities and differences exist between institutions, which may or may not influence the overall quality of an institution's PAM program.

The uncertain taxonomy of [Pasteurella] pneumotropica and other rodent Pasteurellaceae has hindered the acquisition of knowledge on the biology and disease for this group of bacteria. Recently, these organisms have been reclassified within the new genus Rodentibacter. In this study, we documented which of the new described Rodentibacter spp. are present in the mouse and rat microbiologic units of an experimental facility. Screening all of the microbiologic units populated with mice and rats yielded 51 Rodentibacter isolates. Molecular and phenotypic diagnosis indicated the colonization of mice by R. pneumotropicus and R. heylii, whereas R. ratti and R. heylii were found in rats. Overall, we document the association of laboratory rodents with 3 of the newly described Rodentibacter. Diagnostics of the Rodentibacter spp. at the species level can decisively contribute to the progress of knowledge on these bacteria.

Axolotls (Ambystoma mexicanum) from a research colony presented with multifocal, white chalky to gray skin lesions, a diffuse whitish to blue hue to the integument, and friable gill filaments. Skin scrapings and wet mounts revealed Chilodonella, Ichthyobodo, and a trichodinid species. The average overall burden (that is, all 3 species) per axolotl ranged from 0 to 25 parasites per 40 × field (p40f; mean ± 1 SD, 2.6 ± 5.5), with a prevalence of 12%, 60%, and 48%, respectively. Concurrent with husbandry modifications, axolotls were treated with an 8-h static immersion bath that contained 0.025 mL/L 37% formaldehyde. Chilodonella organisms were no longer observed after the initial treatment, and Ichthyobodo decreased from 2.4 ± 5.6 to 0.6 ± 1.8 organisms p40f. However, the average overall burden increased 4-fold to 10.5 ± 9.8 parasites p40f, and the trichodinid organisms increased 13-fold from 0.8 ± 2.3 to 10.4 ± 9.2 organisms p40f. A second treatment consisted of an 8-h immersion bath that contained 0.05 mL/L 37% formaldehyde on 2 consecutive days. A significant change was noted in the average overall burden of 0.5 ± 1.1 parasites p40f, a greater than 5- and 21-fold decrease from pretreatment and after the initial treatment, respectively. No significant change between the first and second treatment was observed for Ichthyobodo, with 0.6 ± 1.2 organisms p40f, but this number represented a significant decrease from pretreatment. After the second treatment, the trichodinid organism was detected in only one axolotl, with a low overall burden of 0.2 ± 0.4 organisms p40f and resulting in a significant decrease in the trichodinid count to 0.01 ± 0.04 organisms p40f. Treatment with formalin (37% formaldehyde), in conjunction with husbandry improvements, was effective in significantly reducing ectoparasite burden and eliminating clinical symptoms in axolotls but did not fully eliminate all protozoa.

Because human patients with monkeypox virus (MPXV) infection report painful symptoms, it is reasonable to assume that animals infected with MPXV experience some degree of pain. Understanding whether and how analgesics affect MPXV disease progression is crucial when planning in vivo challenge experiments. In the current study, we challenged prairie dogs with a low dose (4 ×10³ pfu) of MPXV and treated with meloxicam (NSAID) or buprenorphine (opioid); control animals did not receive analgesia or received analgesia without MPXV challenge. Subsets of animals from each group were serially euthanized during the course of the study. Disease progression and viral kinetics were similar between groups, but MXPVinfected, meloxicam-treated animals showed increasing trends of morbidity and mortality compared with other groups. Differences between no-analgesia MPXV-infected control animals and MPXV-infected animals treated with buprenorphine were minimal. The findings in the current study allow more informed decisions concerning the use of analgesics during experimental MPXV challenge studies, thereby improving animal welfare. In light of these findings, we have modified our pain scale for this animal model to include the use of buprenorphine for pain relief when warranted after MPXV challenge.

Opioids are essential for use in rhesus macaques (Macaca mulatta) that require multimodal analgesia or those unable to receive NSAID as part of their pain management plan. The current opioid epidemic has universally limited the availability of these vital analgesics, compelling clinicians to investigate other options including novel opioid formulations. A commercially available injectable, long-lasting, highly concentrated buprenorphine solution (HCBS) provides therapeutic plasma concentrations lasting 24 h after a single dose in cats ( Felis catus). We hypothesized that this same HCBS would achieve therapeutic concentrations (≥0.1 ng/mL) for at least 24 h in rhesus macaques. In the current study, 6 healthy, adult rhesus macaques were included in a randomized, 2-period, 2-treatment crossover study. The low dose (0.24 mg/kg SC) achieved a peak plasma concentration of 19.1 ± 5.68 ng/mL at 0.308 ± 0.077 h, with an AUC of 236.4 ± 22.5 h/ng/mL and terminal elimination half-life of 19.6 ± 4.02 h; for the high dose (0.72 mg/kg SC), these parameters were 65.2 ± 14.7 ng/mL, 0.034 ± 0.004 h, 641.3 ± 79.4 h/ng/mL, and 20.6 ± 2.30 h, respectively. The mean plasma concentrations for the low and high doses in rhesus macaques significantly exceeded the therapeutic threshold for 48 and 72 h, respectively. One macaque showed mild somnolence at both doses, and another showed mild pruritus at both doses. These findings show that subcutaneous administration of HCBS provides prolonged and long-lasting therapeutic plasma levels for 48 to 72 h dosing without problematic adverse effects and thus represents a potential new analgesic alternative.

Elizabethan collars (E-collars) are commonly used in various species to safeguard healing wounds. However, E-collars inadvertently restrict the expression of normal species-typical behaviors, including coprophagy, self-grooming, and social housing. To maintain social housing in accordance with recommendations in the 8th edition of the Guide for the Care and Use of Laboratory Animals, we implemented the use of human infant pants instead of E-collars for postsurgical protection. We retrospectively reviewed the medical records of 154 intact male New Zealand white rabbits (age, 2 to 3 mo) regarding the use of E-collars (group 1; n = 72) compared with human infant pants (group 2; n = 82) for postoperative protection after 308 femoral angioplasty procedures. Maintenance of social pairs throughout the postoperative phase, replacement rate of infant pants, and self-mutilation rates were measured. Our findings indicate that using infant pants for postoperative protection was most successful in maintaining social housing, offers a more cost-effective option to E-collars, and does not increase the rate of self-mutilation in intact male New Zealand white rabbits.