To protect the biosecurity of research rodent colonies, research institutions frequently require a quarantine period for live animals transferred into their facilities. Quarantine practices often include antibiotic and antiparasitic treatment with drugs such as fenbendazole and macrolide lactones. The influence of these compounds on the resident gut microbiota of mice is unknown, and any effects might subsequently affect model reproducibility. To test the influence of standard quarantine procedures on the composition of the microbiota, C57BL/6 mice, purchased from 2 different commercial suppliers, were randomly assigned to treatment groups (n = 12) by vendor and treated with fenbendazole-supplemented feed, topical moxidectin, both treatments, or no treatment (control), according to our institution's standard treatment regimen and duration. Feces were collected on arrival, immediately after completing the 8-wk treatment, and at 2 and 4 wk after treatment. Fecal DNA was extracted, sequenced, and analyzed to compare the changes in the microbiota of treated and control groups. Although significant main effects of time and treatment and interactions between those variables were detected in comparisons of richness, α-diversity, and β-diversity, the effect sizes associated with any particular treatment were consistently much smaller than that associated with acclimation to a new facility in the absence of any quarantine treatments. This outcome, along with the visual evaluation of principal coordinate analysis based on multiple similarity indices, suggests that time or institution plays a larger role in alterations of the murine gut microbiota than do quarantine treatments on its composition.

Aggression among mice remains a common undesirable problem in laboratory settings, and animal welfare and scientific outcomes may become compromised depending on the severity of aggression. This study evaluated the effect of cage enrichment comprising a bilevel, mounted 'mezzanine' compared with a cotton square or shelter on intracage male aggression over a 6-wk period. Our first study involved home-cage behavioral challenges to male mice from a high-aggression substrain (BALB/cJ) and low-aggression substrain (BALB/cByJ). Aggressive interactions and locomotor activity were scored manually and then compared with measures of activity obtained by using a continuous automated home-cage monitoring system, the Digital Ventilated Caging (DVC) system. BALB/cJ mice exhibited similar levels of aggression across housing conditions, whereas BALB/cByJ mice had lower aggression when housed with a mezzanine. In the second study, videorecordings and continuous DVC automated measures were collected over 24 h and divided into 12-h light and dark phases. BALB/cByJ mice—but not BALB/cJ—mice had increased aggressive behaviors during the dark phase. However, the DVC detected higher activity levels during the dark phase, compared with the light phase, in both substrains. Elevated activity levels recorded by the DVC correlated with fighting bouts and high levels of locomotion. These results show that a bilevel structural form of enrichment reduces aggression, depending on the BALB/c substrain, and confirms higher aggression levels in the BALB/cJ substrain. In addition, our findings provide evidence that the DVC is effective in identifying mouse cages with patterns of high activity levels, signaling possible aggression incidences, thus potentially allowing for early intervention and consequently improving animal welfare.

The emotional state of domestic animals is an essential component of the assessment of their welfare. In addition, sensitivity to various rewards can be a valuable indicator when investigating these states. We aimed to design an exploration test and a contrast test that did not evoke fear and anxiety in C57BL/6N mice but that instead were perceived as positive experiences and that might be used to assess sensitivity to various rewards. The exploratory arena had a larger central area and 8 smaller sections containing various objects. Motivation (measured as latency to enter the arena under conditions of increasing weight of the entrance door), anticipation (measured as latency to enter the arena under conditions of increasing delay in opening the entrance door), and the numbers of visits to the different sections were evaluated during a 5-min session in the arena. In the contrast test, after traversing a runway, half of the mice received a tasty reward (hazelnut cream), whereas the others received a neutral reward (food pellet) at the far end. Latency to reach the reward was recorded. After baseline training, rewards were swapped for half of the mice from each category for 3 d, to establish a negative and positive contrast. Mice were both motivated and showed anticipation to enter the exploration arena; after entering, they were active and visited many sections. In the contrast test, latency during the baseline period was longer for mice given the neutral reward compared with the tasty reward. Compared with baseline, latency during the postshift phase decreased for the positive-contrast group (neutral–tasty reward pattern) but did not differ for the negative-contrast group (tasty–neutral reward pattern). Overall, both tests seemed to be positive experiences for the mice and showed potential for use to investigate reward sensitivity.

Prions are proteinaceous infectious agents that are highly resistant to denaturation. Sterilization of prion-contaminated mouse cages requires chemical agents and increased autoclave temperatures that damage traditional cages, thus increasing facility costs. Disposable cages are a possible alternative that might decrease replacement costs without compromising the environment of the mice. We compared our standard protocol of changing traditional cages and bedding once every 2 wk to an experimental protocol using disposable cages in which only the bedding was changed once every 2 wk over an 8-wk period. We hypothesized that disposable cages would retain an acceptable level of cleanliness (measured by ATP swabs and contact plates) for at least 8 wk when bedding is replaced every 14 d. Results from ATP swabs and contact plates showed no difference between the 2 protocols during the 8-wk experiment. Prolonged use (that is, as long as 8 wk) of disposable cages had no additional environmental concerns, compared with traditional cages.

NSAID analgesics may confound models that require inflammation to mimic disease development in humans. This effect presents a challenge for veterinary staff and investigators, because surgery is often necessary to create mouse models of disease and NSAID are first-line analgesics used to treat postoperative pain. We evaluated robenacoxib, a NSAID highly selective for cyclooxygenase 2, in a carrageenan paw edema (CPE) assay and surgical model of venous thrombosis (VT). We generated a mouse-specific dose–response curve by using the CPE assay for robenacoxib doses of 3.2, 10, 32 and 100 mg/kg SC. Electronic von Frey assay, calipers, and novel software for measuring open-field activity revealed that all robenacoxib doses provided, identified effective analgesia at 3 and 6 h, compared with saline. In addition, the 100-mg/kg dose had measurable antiinflammatory effects but yielded adverse clinical side effects. Because the 32-mg/kg dose was the highest analgesic dose that did not decrease paw swelling, we evaluated it further by using the same nociceptive and behavioral assays in addition to a novel nest-consolidation test, and assessment of blood clotting and hematologic parameters in the surgical VT model. A single preemptive dose of either 32 mg/kg SC robenacoxib or 5 mg/kg SC carprofen protected against secondary hyperalgesia at 24 and 48 h. Neither drug altered clot formation or hematology values in the VT model. The open-field activity software and our novel nest consolidation test both identified significant postoperative discomfort but did not differentiate between saline and analgesia groups. In light of these data, a single preemptive subcutaneous dose of 32 mg/kg of robenacoxib or 5 mg/kg of carprofen did not impede this VT mode but also failed to provide sufficient postoperative analgesia.

The injectable anesthetic mixture ketamine–xylazine is commonly used for electrophysiologic experiments in laboratory animals, especially rodents. General anesthesia can induce significant changes in systemic physiology, including those that compromise neural function, thus introducing research confounds. The extent of such concerns varies by agent. Here in mice, we compared the effects of ketamine–xylazine and urethane–xylazine anesthesia on systemic physiologic parameters and the vestibular sensory evoked potential (VsEP), a tool used commonly to assess peripheral vestibular function. Urethane–xylazine anesthesia provided longer anesthesia, prolonged survival times, and less compromised respiratory and cardiovascular function, compared with ketamine–xylazine. In the absence of countermeasures, mice anesthetized with either ketamine–xylazine or urethane–xylazine showed evidence of hypoxemia and fluctuations in brain temperature, heart rate, respiration rate, and VsEP response latency. The levels of hypoxemia had no effect on VsEP response parameters over the period of study (2 to 5 h). Hypoxemia was effectively countered with O₂ supplementation, which stabilized respiratory rates and improved mean survival times by 160% in mice anesthetized with ketamine–xylazine. Monitoring and controlling brain temperature reduced variation in VsEP latency. VsEP thresholds, latencies, and amplitudes did not differ between mice under ketamine–xylazine compared with urethane–xylazine when the brain temperature was held at the same set point. These findings demonstrate that urethane–xylazine provides improved systemic physiologic conditions during anesthesia in mice and may be substituted for ketamine–xylazine in studies using the VsEP to evaluate peripheral vestibular function. Such advantages may prove useful to research in other neuroscience areas and might reduce the number of animals used to achieve adequate sample sizes.

Anesthetic protocols may influence adrenal function. Effective methods for modulating stress are desirable to minimize secondary effects during the perioperative period. The aim of this study was to evaluate the effects of the administration of propofol with dexmedetomidine or ketamine on corticoadrenal function and heart and respiratory rates. A random treatment-order design was used: each rabbit received all treatments, with at least 14 d between experiments. Rabbits were assigned to 3 treatment groups (10 per group): group 1, 1 mL normal saline solution intravenously; group 2, propofol (3 mg/kg IV) and dexmedetomidine (0.35 mg/kg IM); and group 3, propofol (3 mg/kg IV) and ketamine (1 mg/kg IV). Dexmedetomidine was injected 15 min prior to propofol administration. Blood samples were obtained before drug administration and at 5, 10, 30, and 60 min and 24 h after injection. Serum cortisol and corticosterone levels were measured by competitive enzyme immunoassay. Serum glucocorticoid concentrations did not change in group 2. However, rabbits in group 3 showed an increase in serum cortisol (at 5-60 min) and corticosterone (at 5-120 min) when compared with all other groups at the corresponding time points. This increase probably reflected both propofol- and ketamine-associated stimulatory effects corticoadrenal function. Respiratory rate decreased in groups 2 and 3 animals, and heart rate decreased in group 2, probably due to sympathetic inhibition by propofol and dexmedetomidine. In conclusion, propofol–ketamine provides suitable cardiorespiratory stability in rabbits but enhances glucocorticoid secretion more than dexmedetomidine–propofol anesthesia. Glucocorticoid levels in anesthetized rabbits should be considered during protocol design to minimize the stress response to surgery and to avoid erroneous data interpretation.

Limited information is available regarding the efficacy of opioid analgesics in chinchillas. Here we sought to evaluate the analgesic efficacy and safety of hydromorphone in chinchillas. In a randomized, controlled, blind, complete crossover design, hydromorphone was administered at 0.5, 1, and 2 mg/kg SC to 16 chinchillas. Analgesic efficacy was determined by measuring hindlimb withdrawal latencies after a thermal noxious stimulus (Hargreaves method) at 0, 1, 2, 4, and 8 h after drug administration. Changes in daily food intake and fecal output after hydromorphone administration were recorded. At 2 mg/kg SC, but not at lower dosages, hydromorphone increased withdrawal latencies for less than 4 h. Food intake was reduced after all 3 dosages, and fecal output decreased in the 1- and 2-mg/kg groups. The decreases in these parameters were dose-dependent, with the greatest reduction measured over the first 24 h. Our current results indicate that hydromorphone at 2 mg/kg SC is an effective, short-acting analgesic drug in chinchillas that transiently reduces food intake and fecal output. Further studies are needed to evaluate the safety of hydromorphone in animals undergoing surgical procedures and general anesthesia and to determine whether lower doses provide analgesia in different nociceptive models.

Buprenorphine is routinely used in chinchillas at reported doses of 0.01 to 0.1 mg/kg IM or SC. However, these dose recommendations are based on anecdotal reports or extrapolation from studies in other species. Therefore, the purpose of this study was to evaluate the analgesic efficacy and safety of subcutaneously administered buprenorphine in chinchillas. Using a randomized, blind, controlled, complete crossover design, we evaluated buprenorphine at a single dose of 0.05, 0.1 or 0.2 mg/kg SC (experiment A) and 0.2 mg/kg SC (experiment B). Analgesic efficacy was determined by measuring limb withdrawal latencies in response to a thermal noxious stimulus (Hargreaves method) at 0, 3, 6, 12, and 24 h (experiment A) and at 0, 1, 2, 4, and 8 h (experiment B). In a third experiment, food intake and fecal output were monitored after repeated administration of buprenorphine (0.2 mg/kg SC every 6 h for 3 doses). Buprenorphine at 0.2 mg/kg SC, but not at 0.05 or 0.1 mg/kg SC, significantly increased limb withdrawal latencies for less than 4 h. Self-limiting reduction in food intake and fecal output occurred after administration at the 0.2-mg/kg dose in animals undergoing algesiometry. In chinchillas not undergoing algesiometry, the administration of 3 doses at 0.2 mg/kg SC every 6 h did not reduce food intake but significantly decreased fecal output for the first 24 h. Additional studies are needed to evaluate buprenorphine in different algesiometry models and to establish its pharmacokinetic profile in chinchillas.

Anesthesia can affect measured thyroxine (total T4) concentrations in humans and animals, but its effect in black-tailed prairie dogs (Cynomys ludovicianus) has not yet been studied. We used isoflurane to anesthetize 12 prairie dogs for 60 min. Blood samples were obtained from each animal immediately after anesthesia induction and at 30 and 60 min and used for analysis of plasma T4 concentration. The plasma T4 concentration (mean ± 1 SD) was significantly decreased from baseline (3.49 ± 0.52 μg/dL) at both 30 min (3.24 ± 0.52 μg/dL) and 60 min (3.27 ± 0.65 μg/dL) after induction. Compared with baseline, some of the T4 trends were inconsistent between animals, and individual variability in response was responsible for 86% of the overall variability. Regardless of the observed change under isoflurane anesthesia, all measurements in all prairie dogs and at all time points (2.4 to 4.4 μg/dL) were within the reported normal plasma T4 reference range for this species. In conclusion, isoflurane anesthesia appears to cause a significant but inconsistent reduction in plasma T4 concentrations in black-tailed prairie dogs, but because values remain within normal basal levels, the clinical importance of this effect is likely minimal.

Provision of liquid enteral nutrition (LEN) during the perioperative period is standard practice for rodents undergoing bariatric surgery, yet these diets are associated with several challenges, including coagulation of the liquid diet within the delivery system and decreased postoperative consumption. We investigated the use of a commercially available high-calorie dietary gel supplement (DG) as an alternative food source for mice during the perioperative period. C57BL/6J male mice were fed high-fat diet for 8 to 10 wk prior to surgery. The study groups were: vertical sleeve gastrectomy (VSG) +DG, VSG+LEN, sham surgery+DG, and sham+LEN. Food and water intakes, body weight, and body fat composition was monitored throughout the study. Mice that received DG lost significantly more weight preoperatively than those fed LEN. However, during the postoperative period, body weight, body fat composition, and water and caloric intake were similar among all experimental diet groups. Three mice in the VSG+LEN group were euthanized due to clinical illness during the course of the study. In summary, feeding a high-calorie DG to mice undergoing VSG surgery is a viable alternative to LEN, given that DG does not significantly affect the surgical model of weight loss or result in adverse clinical outcomes. We recommend additional metabolic characterization of DG supplementation to ensure that this novel diet does not confound specific research goals in the murine VSG model.

For delayed dental implantation into the mandible, the implant size should be chosen according to the characteristics of that bone. This study investigated anatomic features of the mandible in beagle dogs, to develop recommendations regarding the correct implantation region and available bone area for delayed dental implantation surgery. We used 20 healthy male beagle dogs to create delayed dental implantation models. The dogs' mandibles underwent cone beam CT (CBCT) imaging; the locations of the middle mental foramen and canine root apex were measured on CBCT images. The dogs then were euthanized and their mandibles measured by using a digital vernier caliper. In addition, the correct implantation region and available bone areas were evaluated. The data obtained by using the 2 measuring methods were compared statistically. The results showed that the positions of the middle mental foramen and canine root apex were relatively fixed, with little variation. The implantation and available bone regions showed little variation among dogs and did not differ significantly between the 2 measuring methods. In conclusion, the correct implantation region (mean ± 1 SD) in the beagle mandible for delayed dental implantation surgery was 17.53 ± 0.46 mm in width. The recommended available bone areas (height × width) were 7.22 ± 0.68 mm × 5.32 ± 0.49 mm (P2), 8.21 ± 0.71 mm × 5.81 ± 0.56 mm (P3), and 9.17 ± 0.65 mm × 6.39 ± 0.56 mm (P4) in the premolar region.