Maintaining compliance with cage density recommendations in The Guide for the Care and Use of Laboratory Animals precludes continuous trio breeding in standard-sized mouse cages. This study evaluated and compared several parameters of reproductive performance, intracage ammonia concentration, and fecal corticosterone levels in 2 strains of mice, C57BL/6J (B6) and B6.129S(Cg)-Stat1^tm1Dlv/J (STAT1^–/–), housed as continuous breeding pairs or trios in standard-sized mouse cages, and continuous breeding trios in standard-sized rat cages. Reproductive performance data indicated that STAT1^–/– trios raised in rat cages weaned significantly more pups per litter than did STAT1^–/– trios raised in mouse cages, and B6 mice had higher pup survival rates at weaning than did STAT1^–/– mice in mouse cages housing continuous breeding trios. In addition, the Production Index was significantly higher for B6 breeding trios in rat cages than for B6 trios in mouse cages. Intracage ammonia concentration increased with cage density, with significantly higher ammonia concentrations in mouse cages housing trios compared with rat cages housing trios. However, fecal corticosterone levels did not differ significantly regardless of genotype, breeding configuration, or cage size, and daily health checks revealed no clinical abnormalities under any of the conditions evaluated. These results suggest that, although continuous trio breeding in standard-sized mouse cages does not seem to compromise mouse welfare, it offers no advantage in reproductive performance compared with pair breeding, and in some cases, it might be disadvantageous in this regard. Further, high intracage ammonia in mouse cages containing breeding trios might necessitate more frequent cage changes.

Research organizations should be proactive in regularly evaluating and refining their animal care and use programs in order to advance animal welfare and minimize distress. Pigs are often used in research, but few empirical studies have examined optimal husbandry and research use practices for pigs in a research environment. We developed the Pig Welfare Working Group (PWWG) to address the need for more formal guidelines on the management and use of pigs in research. The PWWG was a stakeholder focus group whose goal was to identify challenges and opportunities relevant to improving animal welfare through collaboration, knowledge sharing, and inclusive decision-making. Through consensus building, the PWWG developed 12 recommendations for behavioral management, housing, research procedures, transportation, and rehoming programs. The recommendations were rolled out across the contract research organization, business units, sites, and countries. Follow up will be conducted regularly to assess welfare, monitor progress toward implementing the recommendations, and recognize and reward participants making changes at their site.

Infectious agents have varying susceptibilities to thermal inactivation and/or mechanical removal from cages by the use of heated, pressurized water. In this study, we tested whether 5 specific infectious organisms (Candidatus savagella [segmented filamentous bacterium (SFB)], Helicobacter sp., mouse norovirus (MNV), Tritrichomonas sp., and Entamoeba muris) could survive the cage wash process and still infect naïve mice. These 5 organisms were chosen due to their prevalence in rodent colonies, environmental stability, and/or potential to influence experimental outcomes. Cages that had housed mice shedding all 5 organisms were assigned to 1 of 3 treatment groups: 1) sanitization in a tunnel washer followed by autoclaving (121 °C [250 °F] for 20 min; n = 40 cages); 2) sanitization in a tunnel washer (82 °C [180 °F] for an average of 30 s; n = 40 cages); or 3) control (bedding change only; n = 40 cages). The presence of these agents in the cage was assessed by performing PCR on swabs of the empty soiled cage interior before and after the treatment. In addition, to determine if any residual nucleic acid was infectious, 2 Swiss outbred (J:ARC(S)) female mice were housed for 7 d in cages from each treatment group. The above procedures were then repeated so that every week each pair of J:ARC(S) mice ( n = 10 pairs of mice/treatment group) were housed in another cage that underwent the same treatment; this was done for a total of 4 consecutive, 1-wk-long periods. Swabs collected from soiled cages were PCR-positive for SFB, Helicobacter, MNV, Tritrichomonas, and Entamoeba in 99%, 97%, 39%, 63%, and 73% of the cages tested, respectively. Cages in the tunnel wash group that were PCR-positive for SFB, Helicobacter, Tritrichomonas, and Entamoeba before treatment remained PCR-positive in 8%, 15%, 43%, and 10% of positive cages, respectively. None of the cages from the autoclave group were PCR-positive for any of the agents after treatment. None of the mice housed in cages in either the autoclave or tunnel wash groups became infected with any of the agents. However, 80%, 60%, and 100% of the pairs of mice housed in untreated cages were PCR-positive for SFB, MNV, and Entamoeba, respectively. None of the mice housed in untreated cages were positive for Helicobacter or Tritrichomonas. Our results suggest that nucleic acids from these bacterial and protozoal organisms may remain in cages after mechanical cage washing, but these nucleic acids are not infectious, and autoclaving is not necessary to prevent transmission.

After detecting Giardia and Cryptosporidium infections and coinfections in 2 litters of puppies in our vivarium, our team realized that we needed a simple, quick, and economical point-of-care test for concurrent screening of asymptomatic dogs for both organisms. Periodic screening of colony dogs and of all dogs introduced into a colony can prevent the spread of Giardia and Cryptosporidium to immunologically naïve animals and help keep staff safe from these zoonotic organisms. To compare methods for diagnosing Giardia and Cryptosporidium spp. in dogs, we used a convenience sampling of feces from 2 popula- tions of dogs; samples were tested with a lateral-flow assay (QC), a commercially-available direct fluorescent assay (DFA), and an inhouse PCR test using established primers. QC results were analyzed in 2 ways: 1) relative to a reference standard that permitted comparative interpretation of DFA and PCR results; and 2) using Bayesian analysis for comparison independent of a reference standard. The QC test showed good specificity for the detection of Giardia according to both the reference standard (95%) and the Bayesian analysis (98%). Similarly, specificity of the QC for the detection of Cryptosporidium was 95% according to the reference standard and 97% according to Bayesian analysis. However, the sensitivity of the QC test was much lower for both Giardia (reference standard, 38%; Bayesian analysis, 48%) and Cryptosporidium (25% and 40%, respectively). This study demonstrates that the QC test can be used to detect both Giardia and Cryptosporidium in dogs and that positive results can be accepted with confidence, whereas negative tests should be confirmed through secondary testing methods.

Two long-acting formulations of buprenorphine are commercially available as analgesics for rodents. However, these drugs have not yet been studied in nude mice. We sought to investigate whether the manufacturer-recommended or labeled mouse doses of either drug would provide and sustain the purported therapeutic plasma concentration of buprenorphine (1 ng/mL) over 72 h in nude mice and to characterize the injection site histopathology. NU/NU nude and NU/+ heterozygous mice were subcutaneously injected with extended-release buprenorphine polymeric formulation (ER; 1 mg/kg), extendedrelease buprenorphine suspension (XR; 3.25 mg/kg), or saline (2.5 mL/kg). Plasma concentrations of buprenorphine were measured 6, 24, 48, and 72 h after injection. The injection site was examined histologically at 96 h after administration. XR dosing yielded significantly higher plasma buprenorphine concentrations than did ER dosing at every time point in both nude and heterozygous mice. No significant difference in plasma buprenorphine concentrations were detected between nude and heterozygous mice. Both formulations yielded plasma levels of buprenorphine of over 1 ng/mL at 6 h; XR sustained buprenorphine plasma levels above 1 ng/mL for over 48 h, whereas ER sustained this level for over 6 h. Injections sites of both formulations were characterized by a cystic lesion with a fibrous/fibroblastic capsule. ER induced more inflammatory infiltrates than did XR. This study indicates that while both XR and ER are suitable for use in nude mice, XR has a longer duration of likely therapeutic plasma levels and induces less subcutaneous inflammation at the injection site.

Depilatory creams are widely used in research to remove hair in preparation for surgery, imaging, and other procedures. However, few studies have evaluated the effects of these creams on mouse skin. We sought to determine the cutaneous effects of 2 different depilatory formulations of a widely used brand as related to the duration of exposure. We compared a standard body formula [BF] and a facial formula [FF] that is marketed as being more gentle on skin. The cream was applied to one flank for 15, 30, 60, or 120 s; hair on the contralateral flank was clipped and used as a control. Treatment and control skin were scored for gross lesions (erythema, ulceration, and edema), degree of depilation, and histopathologic changes. C57BL/6J (B6) and Crl:CD-1(ICR) (CD-1) mice were used to allow comparison of an inbred/pigmented strain to an outbred/albino strain. BF caused significant cutaneous injury to both strains of mice, whereas FF produced significant cutaneous injury only in CD-1 mice. Both strains showed gross skin erythema, with the most severe erythema seen in CD-1 mice treated with BF. Contact time did not affect histopathologic changes or gross erythema. Both formulations produced depilation comparable to clipping in both strains when left on for a sufficient duration. In CD-1, mice, BF required at least 15 s of exposure, whereas FF required at least 120 s. In B6 mice, BF required at least 30 s of exposure, whereas FF required at least 120 s. The 2 mouse strains did not show statistically significant differences in erythema or histopathologic lesions. Overall, these depilatory creams were comparable to clippers for hair removal from mice but they produce cutaneous injury that may affect research outcomes.

The goals of this study were to determine whether mice would adapt to a low-calorie flavored water gel as their sole source of hydration and whether the addition of acetaminophen, tramadol, meloxicam, or buprenorphine to the gel would affect their intake. Water and gel intakes were measured during a 4-phase study, each of which lasted 1 wk: phase 1, standard water bottle only; phase 2, standard water bottle and a separate tube containing water gel; phase 3, water gel only; and phase 4, water gel containing an analgesic drug. Water consumption, corrected for body mass, was not different between male and female mice when water was available (phases 1 and 2). However, the total consumption of water and water gel was higher for females than males during phase 2, and female mice consumed more gel than males during phase 3. When male and female data were combined, total corrected water intake was not different among the first 3 phases of the study. Gel intake did not change significantly after the addition of acetaminophen, meloxicam, buprenorphine or tramadol as compared with untreated water gel. These data suggest that drugs presented in the low-calorie flavored water gel may be a viable alternative to injection or gavage for the administration of analgesic drugs.

Although mice are social animals, individual housing is sometimes requested after surgery. We questioned whether pair-housing mice after surgery resulted in greater trauma to the surgical site as compared with single housing. We further evaluated the effect of individual housing after surgery on the wellbeing of mice that had previously been pair-housed. Female C57Bl/6 mice (age, 6 to 8 wk) were housed as follows: group A, individually housed before and after surgery (n = 10; all 10 mice underwent surgery); group B, pair-housed before surgery but individually housed after surgery (n = 10; all 10 mice received surgery); group C, pair-housed before and after surgery (n = 20; 10 mice underwent surgery but their cage mates did not); and group D, pair-housed before and after surgery (n = 10; all 10 mice underwent surgery). Dependent variables were body weight, body condition, grimace based on real-time scoring, nest building, time to incorporate into nest test (TINT) score, wound trauma score, and missing wound clips. Weight was significantly different between groups A and C both before and after surgery. Mean nest building scores were significantly higher for pair-housed (groups C and D) than for individually housed mice (groups A and B) after surgery while TINT scores were significantly higher for these same groups both before and after surgery. Mean values for body condition, grimace score, wound score, and number of wound clips missing did not differ significantly between any groups either before or after surgery. Taken together, these results suggest that pair housing mice after surgery benefited their wellbeing but did not increase trauma to the surgical incision site or disturb wound clips as compared with individually housed mice. Furthermore, separating previously pair-housed mice (group B) did not affect these measures as compared with individually housed mice (Group A) either before or after surgery.

Nasal drug delivery in rodents is a challenging procedure, especially for brain targeting, as the position of the material in the nasal cavity determines the success of the administration method. The objective of this study was to assess a novel intranasal administration technique for nose-to-brain delivery of biodegradable nasal films. The method was performed in C57BL/6 (n = 10; age, 8 wk) under inhaled sevoflurane. Twenty-four gauge catheters were used for the procedure. Hydroxypropyl methyl-cellulosebased film was formed in the lumen of the catheter and then delivered into the mouse nostril by pushing it out of the lumen using a trimmed and polished needle. Methylene blue was incorporated in the film-forming gel to indicate the delivery area in which the films were deposited. After administration, all mice recovered from anesthesia without incident. None of the mice showed any signs of injury, discomfort, or nose bleeding, thus allowing us to characterize the administration method as noninvasive. Furthermore, postmortem evaluation revealed olfactory-centered placement of the polymeric films, confirming the accuracy and repeatability of the method. In conclusion, this study documented the use of, a novel, noninvasive, intranasal administration technique for nose-to-brain drug delivery in biodegradable films for use in mice.

Barbiturate overdose is a common method for euthanizing pigs. However, barbiturates can cause tissue damage and may affect experimental results, so the minimal dose should be used. The minimal dose of barbiturate for euthanasia in pigs under isoflurane anesthesia has not yet been determined. In this study, we compared the effect of low and high doses of 2 barbiturates (pentobarbital, 30 or 60 mg/kg; thiopental, 20 and 40 mg/kg) on hemodynamic parameters and time to cardiac arrest in female pigs maintained on isoflurane. Acute decreases in blood pressure and end-tidal CO₂ occurred in all pigs shortly after administration of the barbiturate. However, these changes were not different between either of the high- and low dose groups. Cardiac arrest occurred significantly faster for high dose as compared with low dose thiopental groups, but this parameter was different between the 2 pentobarbital groups. The bispectral index fell immediately after dosing, in all pigs, but no significant differences were observed in the time needed to achieve 0 for the high or low-doses of either drug. In pigs maintained on isoflurane, a low dose of barbiturates is adequate for euthanasia and may result in less tissue damage.