IACUCs serve a critical role in animal care and use programs, ensuring that institutions which use animals in research and teaching do so responsibly and humanely. This role is defined in part by federal regulations, policies, and guidelines that prescribe the establishment and function of these committees. Often, IACUC administrators are expected to evaluate IACUC performance to ensure that committees execute these functions effectively, and in a manner that is suitable to the institution. However, methods for IACUC performance evaluation have not been well described in the peer-reviewed literature. To address this deficit, we conducted a systematic review using MEDLINE to identify methods that have been used to assess IACUCs. The scope of this review was intentionally broad to capture evaluation methods used by other institutional committees with similar responsibilities in overseeing research conduct, including animal ethics committees (AECs), institutional biosafety committees (IBCs), and institutional review boards (IRBs). Over 100 publications that included empirical evaluation methods were identified, although only 17 evaluated IACUCs in the United States. A substantial number of the studies used qualitative methods, such as surveys or questionnaires, interviews, and observations. The IACUC functions and characteristics most often assessed in the 17 publications included components of the protocol review processes and committee membership. We compiled this information to offer IACUC administrators a source of methodologies that can be incorporated into quality improvement and IACUC performance evaluation efforts. We also suggest ways in which organizations may evaluate IACUCs using methods described in the literature for other types of committees.

Noise and vibration are present in every room of laboratory animal vivaria, with great variability from room-to-room and facility-to-facility. Such stimuli are rarely measured. As a result, the many stakeholders involved in biomedical research, (for example, funding agencies, construction personnel, equipment manufacturers, animal facility administrators, veterinarians, technicians, and scientists) have little awareness of the effects such stimuli may have on their research animals. Noise and vibration present a potential source of unrecognized animal distress, and a significant, uncontrolled and confounding variable in scientific studies. Unmeasured and unrecognized noise and vibration can therefore undermine the fundamental goals of the 3R's to refine animal models and reduce the number of animals used in biomedical and behavioral research. This overview serves to highlight the scope of this problem and proposes a series of recommended practices to limit its negative effects on research animals and the scientific data derived from them. These practices consist of developing a written plan for managing noise and vibration concerns, assessment of noise and vibration both annually and whenever unexpected changes in the facility or animals are observed, and for maintaining levels of chronic noise below thresholds that might cause animal welfare concerns or disruptions in ongoing studies.

Several diagnostic tools are available for veterinarians and fish health professionals to evaluate fish health and their abnormalities. However, reference data regarding the character and size of fish blood cells are limited. The purpose of this study was to analyze the morphology and morphometry of normal blood cells in zebrafish (Danio rerio), common carp (Cyprinus carpio carpio), and tilapia (Oreochromis niloticus). To get a representative sample, we took blood from ten 6-mo old healthy fish from each species. Fish were purchased from an ornamental fish market and the local government fish breeding center in Yogyakarta, Indonesia. A total of 100 erythrocytes and a maximum of 30 leukocytes (neutrophils, eosinophils, basophils, lymphocytes and monocytes) were randomly sampled and analyzed qualitatively and quantitatively, including morphometric analysis of both the long axis (LA) and short axis (SA) of these cells. All data obtained were analyzed using ANOVA statistical tests to compare the blood cells in each species with SPSS software. The findings revealed distinct differences in both morphology and morphometry of the blood cells among the species. Basic knowledge obtained from this research will aid in the development of biomarkers and other ancillary diagnostic tools for further hematology research, conservation, and clinical diagnosis in these 3 fish species.

The objective of this study was to optimize cryopreservation of sperm from Mauritian cynomolgus macaques (MCM) in defined conditions. Sperm viability and motility were compared between sperm cryopreserved in chemically-defined freezing media with variable osmolarity and the presence of either ethylene glycol or glycerol. The highest percentage viability (after freeze-thaw) was seen in sperm samples that were cryopreserved in medium with an osmolarity of 310 mOsm, while higher osmolarities markedly decreased sperm viability. Ethylene glycol and glycerol at concentrations of 4.6% and 5%, respectively, preserved sperm viability to an equivalent degree. Although higher motility rates and higher straight-line velocities were observed in sperm samples frozen in glycerol compared with ethylene glycol, these differences were not statistically significant. Thawed sperm frozen in defined conditions with glycerol were capable of fertilizing MCM oocytes in vitro, with development to the blastocyst stage. The protocol described here provides an effective method for cryopreservation of sperm to facilitate subsequent in vitro fertilization and genome editing of embryos in MCM species.

Control mice housed in the same room as mice with pancreatic ductal adenocarcinoma (PDAC) demonstrate decreased food intake coincident with the cachexia experienced by the mice with PDAC. Mice are considered an empathetic species, and we hypothesized that the reduced food intake in normal mice was an "empathy state" that was mediated by olfactory cues. Naïve male and female C57BL/6 mice were exposed to soiled bedding from mice experiencing PDAC induced cachexia or from control mice in the PDAC study. Body weight, food intake, and food spillage were measured across 48 h. Statistically significant differences in food consumption were found at various time points in both positive and negative directions for the 2 bedding conditions, and the direction of effect was opposite for males and females. Although analysis of data from previous PDAC studies showed differences in food spillage between PDAC mice and their controls, in this study we found no correlation between food consumption and food spillage based on bedding type. Disruption of food intake due to the "empathy state" requires testing larger numbers of animals to attain appropriate statistical power, which is contrary to the goal of using fewer animals. Empathy effects require careful consideration of sample size and cautious interpretation of results. This study also highlights the importance of sex as a biologic variable and why quantifying food spillage is important in studies of food intake.

Corncob is a common bedding material used in laboratory rodents, but little is known about differences in the effects of the 2 available sizes on rodent models and health. This study compared the effects of these 2 corncob bedding sizes on cage ammonia levels, behavior, and respiratory pathology in mice. We hypothesized that the beddings would not differ significantly in their effects on these parameters. Two strains of male mice (C57BL/6 and 129S1/Svlm) were housed in static, filter-top cages containing 1 of the 2 bedding types for the duration of the study (12 wk). Intracage ammonia was measured during 1 wk of the study on days 0, 3, 5, and 7. Behavior was evaluated by using circadian rhythm, open field, and Morris water-maze tests. Animals were euthanized with injectable euthanasia solution to collect respiratory and ocular tissues for histopathologic lesion scoring. Animals that were euthanized immediately upon arrival from the vendor served as negative controls. Bedding size did not significantly affect behavior or ammonia levels. Average intracage ammonia levels on day 7 were 525 ppm for 1/4-in. bedding and 533 ppm for 1/8-in. bedding. Regardless of the bedding size, lesions noted in both strains of mice were of similar incidence and severity, were limited to the nose, and consisted of minimal to mild suppurative rhinitis. The eyes, trachea, and lungs were not affected. In conclusion, 1/4-in. and 1/8-in. corncob beddings have comparable effects on cage ammonia levels and the behavior and respiratory pathology in male mice of the strains tested.

Hydrogen peroxide (HP) decontamination is effective for a wide spectrum of pathogenic microorganisms. However, exposure to HP causes deleterious effects on some materials. The purpose of this study was to examine material compatibilities with ionized and vaporized hydrogen peroxide (iHP and VHP). With regard to iHP, 24 kinds of materials were exposed up to 100 cycles to iHP. The tested materials included plastics, metals, woods and plated or coated goods. The procedure of iHP decontamination was as following: gas time (11 min), dwell time (15 min) and aeration time (120 min). iHP decontamination caused some damage to copper, brass, chromium plate and galvanized iron immediately after exposure. Repeated iHP decontamination caused marked damage in stainless steel and urethane-, silicone- or epoxy-coating materials. Condensation of iHP decontamination posed severe damage for the material surfaces. With regard to VHP, 36 kinds of materials were exposed for up to 200 cycles to VHP decontamination. Under dry (dehumidified) conditions, VHP decontamination caused few changes on the surfaces of resin materials in dry conditions, although some resins began to develop hardening or softening. Discoloration was found in the stainless steel and changes in its coating materials. Bleaching was also observed in wooden materials. Under condensation conditions of VHP, nylon softened and butyl rubber hardened. Condensation of VHP caused material damage such as discoloration in the stainless steel, corrosion of zinc-plated steel, and air-bubbling under the color-steel sheet. The high concentrations of HP with condensation caused severe changes in metals and resins after repeated exposure. The VHP decontamination tests provided evidence that the material damage was more severe under condensation conditions than under dry conditions. Our results demonstrate the importance of condensation of HP when using it to decontaminate equipment.

Current methods for eradicating Corynebacterium bovis, such as depopulation, embryo transfer, and cesarean rederivation followed by cross fostering, are expensive, complex, and time-consuming. We investigated a novel method to produce immunocompromised offspring free of C. bovis from infected NOD. Cg-Prkdc^scidIl2rg^tm1Wgl/SzJ (NSG) breeding pairs. Adult NSG mice were infected with C. bovis, paired, and randomly assigned to either a no-antibiotic control group (NAB, n = 8) or a group that received amoxicillin–clavulanic acid (0.375 mg/mL) in their drinking water for a mean duration of 7 wk (AB group, n = 7), spanning the time from pairing of breeders to weaning of litters. The AB group also underwent weekly cage changes for 3 wk after pairing to decrease intracage C. bovis contamination, whereas the NAB mice received bi-weekly cage changes. Antibiotics were withdrawn at the time of weaning. All litters (n = 7) in the AB group were culture- and qPCR-negative for C. bovis and remained negative for the duration of the study, whereas all litters in the NAB group (n = 6) remained C. bovis positive. A single adult from each breeding pair was sampled at weaning and at 5 and 10 wk after weaning to confirm the maintenance of (NAB) or to diagnose the reemergence (AB) of C. bovis infection. By the end of the study, C. bovis infection had returned in 3 of the 7 (43%) tested AB adults. Our data suggest that metaphylactic antibiotic use can decrease viable C. bovis organisms from adult breeder mice and protect offspring from infection. However, using antibiotics with frequent cage changing negatively affected breeding performance. Nevertheless, this technique can be used to produce C. bovis-free NSG offspring from infected adults and may be an option for salvaging infected immunocompromised strains of mice that are not easily replaced.

Accurate pain assessment methods are necessary to ensure animal welfare and reliable data collection in animal research. The Rat Grimace Scale (RGS), a facial expression pain scale, allows effective identification of pain. However, the potential confounds of this method remain mostly unexplored. General anesthesia, which is used in many laboratory procedures, suppresses thermoregulation and results in hypothermia. We investigated the effects of isoflurane-induced hypothermia on RGS scores. Twenty (10 male and 10 female) Sprague–Dawley rats each received 30 min of anesthesia, followed by 30 min of observation after the return of sternal recumbency. Rats were randomized to receive warming with an electric heating pad or no warming during both periods. Unwarmed rats became hypothermic within 15 min after isoflurane exposure began and returned to normothermia within 15 min after returning to sternal recumbency. Warmed rats did not deviate from the normothermic range. The RGS scores of unwarmed rats were significantly higher than baseline levels for 3 h after anesthesia and were higher than those of warmed rats at 5 and 180 min after anesthesia. Hypothermia resulted in a larger proportion of rats crossing a predetermined analgesic intervention threshold. Our findings show that hypothermia induced by isoflurane anesthesia presents a confound to accurate RGS scoring. These results emphasize the importance of maintaining normothermia to avoid inflated pain scores and to obtain accurate pain assessment.

An extended-release formulation of the NSAID meloxicam (MSR) is used to provide 72 h of continuous analgesia in many species, including rodents. Although standard formulations of meloxicam are frequently used in rats with no observable injection-site reactions, the potential adverse effects from MSR have not been characterized sufficiently nor has a prospective study of these effects been performed in rats. To address this deficiency, we evaluated injection-site reactions after a single subcutaneous administration of MSR (n = 16) or sterile saline (SC, n = 6) in the flank of age- and sex-matched Sprague–Dawley rats. Mass and erythema scores were measured daily for 2 wk, and injection sites were collected for histopathology after euthanasia. Rats were randomly selected for euthanasia at 7 d (n = 12) or 14 d (n = 10) after injection to capture the subacute and chronic phases of mass and erythematic lesion formation. No rats in the SC group developed lesions, whereas all 16 MSR-treated rats developed masses. The median time to first mass in the MSR treatment group was 3 d (95% CI, 2–3 d), and nearly 8 d for erythema (95% CI, 6.7–9.1 d). The trajectory of mass lesion severity showed rapid progression from score 1 at onset (day 2 or 3) to score 2 for almost all animals by day 5 or 6. Histopathology was characterized by localized inflammation with central necrosis and peripheral fibrosis, with some sections showing developing draining tracts. Given the high prevalence and severity of localized skin reactions, MSR analgesia should be considered carefully for Sprague–Dawley rats.

The purpose of this study was to compare the safety and efficacy of buffered tricaine methanesulfonate (MS-222) and isoeugenol for the anesthesia of zebrafish undergoing caudal fin clipping. Eighty 9 mo Danio rerio (AB strain) zebrafish were allocated to one of 2 equal groups: buffered MS-222 (168 mg/L, n = 40) or isoeugenol (20 mg/L, n = 40). The time to induction of anesthesia was significantly shorter in the isoeugenol group (141 ± 70 s) than in the MS-222 group (207 ± 103 s). The time to recovery from anesthesia was also shorter in the MS-222 group (373 ± 125 s) than in the isoeugenol group (491 ± 176 s). No obvious displays of distress or aversion to anesthesia were observed in either group. No difference was detected in the proportion of zebrafish that became anesthetized with either drug. One male zebrafish in the buffered MS-222 group was found dead at the 1-h post-procedural monitoring time point, but there was no difference between groups in the proportion of fish that survived anesthesia to the end of experiment. In conclusion, the safety and efficacy of buffered MS-222 (168 mg/L) and isoeugenol (20 mg/L) was similar for zebrafish undergoing anesthesia for caudal fin clipping.

In cynomolgus macaques, plasma levels of sustained-release formulations of meloxicam meet or exceed efficacious concentrations for 48 to 72 h, thereby allowing less animal handling and providing more consistent efficacy than standard formulations of meloxicam. The goal of this study was to compare the pharmacokinetics of a single subcutaneous dose of a sustained-release formulation of meloxicam (Melox-SR) with those of oral (Melox-PO) and standard subcutaneous (Melox-SC) formulations dosed every 24 h for 3 consecutive days. Dogs (5 or 6 adult male Beagles) each received the following 3 treat- ments: first, Melox-SR (10 mg/mL, 0.6 mg/kg SC once), next Melox-SC (0.2 mg/kg SC once, followed by 0.1 mg/kg SC every 24 h), and finally Melox-PO (same dosage as Melox-SC), with a washout period of at least 2 wk between formulations. Blood was collected at 0 (baseline), 1, 4, 8, 12, 24, 48, and 72 h after the initial administration of each formulation for comparison of meloxicam plasma concentrations. Blood was also collected before administration and at 48 h after Melox-SR injection for CBC and chemistry analysis. Plasma concentrations (mean ± 1 SD) of Melox-SR peaked at the 1-h time point (2180 ± 359 ng/ mL), whereas those of Melox-PO (295 ± 55 ng/mL) and Melox-SC (551 ± 112 ng/mL) peaked at the 4-h time point. Melox-SR yielded significantly higher plasma concentrations than Melox-PO and Melox-SC until the 48 and 72-h time points, respec- tively. Melox-SC plasma concentrations were significantly higher than those of Melox-PO at 4, 8, 12, 24, 48 and 72 h. No lesions were noted at the Melox-SR injection sites, and Melox-SR administration was not associated with changes in the CBC and serum chemistry panels. A single 0.6-mg/kg dose of Melox-SR can yield plasma concentrations that exceed 350 ng/mL for at least 72 h in adult male dogs.

Euthanasia is a necessary component in research and must be conducted humanely. Currently, regulated CO₂ exposure in conscious rats is acceptable, but data are divided on whether CO₂ alone is more distressing than anesthesia prior to CO₂. To evaluate distress in rats, we compared physiologic responses to CO₂ euthanasia with and without isoflurane preanesthesia. Male Sprague–Dawley rats were implanted with telemetry devices to measure mean arterial pressure (MAP), heart rate (HR), and blood glucose. Animals recovered for 2 wk and were then exposed to either 5% isoflurane (n = 6) or 100% CO₂ (n = 7; calculated 30% chamber volume/min displacement) in their home cages to induce loss of consciousness. Euthanasia was then completed with CO₂ in both groups. MAP and HR increased when the gas delivery lids were placed on the home cages of both groups. Both MAP and HR gradually decreased with isoflurane exposure. MAP increased and HR decreased with CO₂ exposure. Glucose levels remained stable throughout the procedure, except for a small drop in conscious animals initially exposed to 100% CO₂. These data suggest that both gases affect the measured parameters in a similar manner, and that environmental factors, such as gas delivery lid placement, also change these measurements.