Some performance standards for continuous trio breeding in 'shoebox' cages for inbred stocks and outbred strains of mice challenge the minimum floor space recommendations in the 8th edition of the Guide for the Care and Use of Laboratory Animals. In our study, we evaluated whether continuous trio breeding could be successfully applied to a breeding colony of genetically engineered mice housed in shoebox cages with a floor area of 67.6 in². Mice heterozygous for genetically engineered mutations to estrogen receptors and their wildtype counterparts were continuously bred as trios or pairs. Confounding environmental factors were controlled through standardized husbandry practices and husbandry, and all mice were bred simultaneously to control for temporal factors. Several measures of reproductive performance—including number of litters per female, production index, interlitter interval, litter size at birth, litter size at weaning, weaning rate, and body weight of pups at weaning— were evaluated over approximately 6 mo. Regardless of genotype, interlitter interval, litter size at birth, and litter size at weaning were significantly lower for trio-bred mice than for pair-bred mice. In addition, significant interactions emerged between genotype and breeding strategy for these reproductive measures. Furthermore, significant differences between genotypes occurred for interlitter interval and weaning rate, regardless of breeding strategy. Underlying mechanisms to account for effects of genotype on interlitter interval and the interaction of genotype with breeding strategy were unclear but may reflect effects of overcrowding and reproductive suppression.

Cleaning behavioral equipment between rodent subjects is important to prevent disease transmission and reduce odor cues from previous subjects. However, the reporting regarding the cleansing procedures used during such experiments is sporadic and often incomplete. In addition, some investigators are reluctant to clean devices between subjects because they are concerned that animals will react negatively to the smell of the cleansing agents. We hypothesized that mice tested on an elevated plus maze (EPM) soiled with excretions from conspecifics would test as being more stressed than mice tested on the same apparatus that was cleaned between animals. We tested the performance of C57BL/6J mice on an EPM sanitized with 3 common cleaning agents—isopropyl alcohol, chlorine dioxide, and bleach—and on an EPM soiled with rodent urine, feces, and presumably pheromones. We further tested the potentially aversive nature of the cleansing agents by using the classic light:dark box and a 2-choice light:dark box. Our data indicate that cleaning the EPM compared with leaving it soiled did not affect performance in male or female C57 mice, nor did cleaning agent choice. In addition, test subjects did not react to the presence of the cleaning agents when incorporated into the classic light:dark test. However, in the 2-choice light:dark test, mice given the option to avoid an area containing a cleaning agent showed aversion to all 3 agents, when all other conditions were equal. Given the lack of an observable effect of cleaning on EPM performance, we recommend cleaning of the EPM device between C57 mice to minimize the potential spread of disease.

The provision of nesting material benefits mice by reducing cold stress, improving feed conversion, increasing litter size, and improving adaptive immunity. The effects of toxins are sensitive to environmental changes, and the introduction of novel items can alter results in some toxicologic studies. We hypothesized that nesting material would reduce stress and positively alter immunologic parameters in Crl:CD1(ICR) mice, thus changing typical results from a well-studied immunomodulating drug, cyclophosphamide. A 13-wk study assessed the following treatments in a factorial design (n = 4; 32 cages total): nesting (0 or 10 g) and drug (50 mg/kg cyclophosphamide or 10 mL/kg saline; IP weekly). Detailed examinations and body weights were recorded weekly, and nests were scored twice weekly. Fecal pellets were collected at 0, 4, 6, and 12 wk for analysis of corticosterone metabolites. At study termination, clinical pathology and immune parameters were collected, a necropsy performed, and lymphoid organs and adrenal glands were submitted for histopathology. All expected results due to cyclophosphamide were observed. Nesting reduced the proportion of mice with piloerection, and body weights were highest in saline–nested male mice. No differences in hematology, clinical chemistry, or absolute lymphocyte counts were observed. Corticosterone metabolites in all nested groups were not different from baseline levels but all nonnested groups had higher levels than baseline. Nested cyclophosphamide-treated groups had significantly lower corticosterone levels than nonnested cyclophosphamide-treated groups. This study illustrates that nesting material does not alter the results of a standard toxicology study of cyclophosphamide but alleviates study-related stress and improves mouse welfare.

Swine (Sus scrofa) are often the 'gold standard' laboratory animal for ophthalmology research due to the anatomic and physiologic similarities between the porcine and human eye and retina. Despite the importance of this model, few tools for behavioral vision assessment in pigs are available. The aim of this study was to identify and validate a feasible and reproducible behavioral test to assess vision in a pig model of photoreceptor degeneration. In addition, a robust behavioral test will reduce stress and enhance enrichment by allowing animals opportunities for environmental exploration and by reducing the number of invasive experimental procedures. Two distinct behavioral approaches were tested: the obstacle-course test and temperament test. In the obstacle-course test, pigs were challenged (after an initial training period) to navigate a 10-object obstacle course; time and the number of collisions with the objects were recorded. In the temperament test, the time needed for pigs to complete 3 different tasks (human-approach, novel-object, and open-door tests) was recorded. The obstacle-course test revealed significant differences in time and number of collisions between swine with vision impairment and control animals, and the training period proved to be pivotal to avoid bias due to individual animal characteristics. In contrast, the temperament test was not altered by vision impairment but was validated to measure stress and behavioral alterations in laboratory pigs undergoing experimental procedures, thus achieving marked refinement of the study.

Because tetanus can cause significant morbidity and mortality in NHP, colonywide vaccination with tetanus toxoid is recommended for outdoor breeding colonies of rhesus macaques, with primary immunizations commonly given to infants at 6 mo of age followed by booster vaccines every 10 y. Maternal antibodies are thought to offer protective immunity to infants younger than 6 mo. However, historical colony data from the Yerkes National Primate Research Center show a higher incidence of tetanus among infants (≤ 6 mo old) born to subordinate dams. Whether this higher incidence of infantile tetanus is due to a higher incidence of trauma among subordinate animals or is a stress-induced impairment of maternal antibody protection is unknown. Studies in other NHP species suggest that chronic exposure to social stressors interferes with the receptor-mediated transplacental transfer of IgG. Therefore, the primary aim of this study was to determine whether chronic stress associated with social subordination impairs prenatal transfer of antitetanus immunity in breeding female rhesus macaques. Subjects included 26 high- and 26 low-ranking adult female rhesus macaques that were nearly 5 or 10 y after their initial immunization and their nonimmunized infants. We hypothesized that infants born to subordinate dams that were nearly 10 y after immunization would have the lowest infant-to-dam antibody ratios and thus would be at greatest risk for infection. Results revealed no significant intergroup differences in infant antitetanus IgG levels. However, infant-to-dam IgG ratios against tetanus were significantly lower among subordinate animals compared with dominant macaques, after accounting for the number of years since the dam's initial vaccination. In addition, higher maternal hair cortisol levels predicted lower infantto-dam tetanus toxoid IgG ratios. Together, these findings suggest that chronic social stress in female rhesus macaques may hamper the prenatal transfer of antitetanus immunity to offspring.

This study investigated the analgesic activity of tramadol in female C57BL/6J mice by using a single subcutaneous injection (25 mg/kg) of tramadol combined with the same dose given in drinking water for 24 h. We then evaluated the pharmacokinetics of tramadol and its active metabolite O-demethyltramadol (M1). To evaluate pain and analgesic efficacy, we performed clinical and behavioral assessment, burrowing tests, and activity analysis and measured body weight, food and water intake in mice that were untreated (control) or underwent analgesia only (T); anesthesia and surgery (AS); or anesthesia, surgery, and analgesia (AS+T). The plasma concentration of tramadol decreased rapidly whereas, for more than 18 h, the M1 level remained stable and above its minimal analgesic concentration for humans. Total food and water intake over 24 h was comparable among all groups. Although T mice consumed tramadol-treated water in sufficient amount and frequency, AS and AS+T animals showed decreased drinking frequency during the first 4 h after surgery. Compared with control and T groups, composite pain scores and burrowing latencies increased significantly in both AS and AS+T mice after surgery, suggesting postsurgical pain. However, AS and AS+T mice did not differ significantly after surgery. In conclusion, although naïve animals ingested a sufficient amount of the drug and plasma levels appeared sufficiently high, mice markedly reduced water intake immediately after surgery. Consequently, even in combination with an initial drug injection, the subsequent voluntary tramadol intake was insufficient to reduce signs of postsurgical pain significantly after laparotomy.

Various anesthetic protocols are used in laboratory swine, each with specific advantages and disadvantages. Partial intravenous anesthetic techniques (PIVA) help minimize dose-dependent cardiopulmonary effects of inhalant drugs. The aim of this study was to determine the cardiopulmonary effects of a PIVA in laboratory swine. In a prospective, nonrandomized clinical study, 8 healthy juvenile Landrace–White pigs were premedicated with azaperone (0.20 ± 0.20 mg/kg IM), dexmedetomidine (0.02 ± 0.002 mg/kg IM), and alfaxalone (2.0 ± 0.20 mg/kg IM), and anesthesia was induced with intravenous alfaxalone. Anesthesia was maintained by using constant-rate infusion of dexmedetomidine (2 μg/kg/h) and alfaxalone (25 μg/kg/min) in combination with isoflurane. After the fraction of expired isoflurane was adjusted to 1.1% to 1.5%, respiratory rate, heart rate, systemic and pulmonary arterial pressure, central venous pressure, cardiac output, bispectral index, systemic vascular resistance, and arterial and mixed venous blood gases were recorded every 10 min for 60 min. Statistical analysis consisted of repeated-measures one-way ANOVA. Significant decreases occurred in heart rate, pulmonary mean arterial pressure, pulmonary diastolic pressure, partial pressure of arterial oxygen, partial pressure of venous oxygen; significant increases occurred in respiratory rate, minute volume index, diastolic arterial blood pressure, systemic vascular resistance, and arterial pH over time. We consider that the observed statistically significant cardiopulmonary changes were clinically important and that the PIVA protocol provided hemodynamic and respiratory stability for short-term anesthesia of laboratory swine.

Many biomedical research protocols for mouse models involve serial blood collection and analysis. Two common techniques for serial blood collection in this species are the retrobulbar (RB, also called retroorbital) and facial vein (FV) methods. However, previous studies comparing these methods typically evaluated collection at a maximum of 2 time points. Here we compared hematologic values, adverse clinical effects, and histopathologic lesions in mice bled either once or serially (6 times) by using the FV or RB method. Mice (n = 48) were divided into 4 groups: single FV, single RB, serial FV and serial RB. Mice in the single-collection groups underwent a single blood collection by the indicated method, whereas those in the serial-collection groups were sampled once weekly for 6 consecutive weeks. All animals were euthanized and necropsied 2 wk after their last blood collection. Compared with all other groups, the serial FV group experienced more serious clinical adverse events, including 33% mortality, convulsions, head tilt, and hemorrhage from the ear canal and nares. In addition, mice in the FV groups had a significantly greater acute body weight loss compared with mice in the RB groups. Histologically, mice in both serial-collection groups had an increased incidence of tissue lesions compared with their respective single-collection groups. Importantly, only mice in the serial FV group had life-threatening histopathologic lesions, including cerebral hemorrhage or ischemia. Given these data, we conclude that serial blood collection in mice causes increased incidence of tissue damage compared with single sampling, and serial blood collection by the FV method causes substantial morbidity and mortality compared with the RB method.

Laboratory mice (Mus musculus) are susceptible to hypothermia, especially during anesthetic events, disease states, and exposure to environmental stressors. Thermal support devices for small mammals are numerous, but often require a power source and may be impractical to use for cages on a rack. Air-activated thermal devices (AATD) are mixtures of chemicals that cause an exothermic reaction. In this study, we examined the environmental effects of AATD on internal cage temperatures without the use of additional equipment as well as the physiologic effects of AATD as postoperative thermal support in mice. For environmental experiments, temperatures measured inside the cage and above the AATD peaked at 35.6 ± 2.5 °C (13.4 °C higher than control cages). We also demonstrated that the amount of heat produced by AATD and its temporal distribution are dependent on cage and rack types. For physiologic experiments, mice were surgically implanted with an intraperitoneal temperature telemetry device in a static cage setting. Recovery times and final body temperature at 5 h postoperatively did not differ significantly between mice with and without AATD. During the first 0 to 3 h after mice returned to their home cages, body temperature dropped markedly in mice without AATD but not in mice with AATD. Based on this result the physiologic results of our study support that AATD can be useful in providing extended thermal support for mice housed in static microisolation cages to help maintain body temperature postsurgically. Environmental results of our studies demonstrated that AATD provide local clinically relevant thermal support for 2.5 to 6 h, depending on cage set-up.

Preparing the skin of rodents for surgery often involves multiple applications of antiseptic agents. However, fewer applications may achieve the same antiseptic outcome. We evaluated the antimicrobial efficacy and effects on intraoperative body temperature of various surgical scrub agents, including novel waterless alcohol-based (WAB) options. Prior to ventral laparotomy, female C57BL/6 mice were treated with 0.9% saline (control); 70% ethanol; 10% povidone–iodine alternated with saline or 70% ethanol; 2% chlorhexidine digluconate alternated with saline or 70% ethanol; or 1 of 3 WAB products—commercial surgical scrub A, commercial surgical scrub B, or a common commercial hand sanitizer. Core temperatures were recorded, and aerobic culture swabs were collected from the surgical site at multiple time points. Intraoperative temperature trajectories for animals treated with scrub B, 10% povidone–iodine with saline, or hand sanitizer did not differ from saline (control). Temperature trajectories of mice treated with other scrub agents did differ significantly from saline. Bacteria were not detected at the operative site after 3 scrubs of 70% ethanol or 10% povidone–iodine alternated with ethanol, 2 scrubs of scrub A or B, 1 scrub of hand sanitizer, and both 1 and 3 scrubs of 2% chlorhexidine alternated with ethanol. Scrub B and 2% chlorhexidine–ethanol demonstrated prolonged antibacterial efficacy. Histology of corresponding haired skin sections revealed no differences in postoperative healing between groups, and no postoperative infections occurred. These results indicate that various novel WAB disinfectants, particularly scrub B (61% ethanol and 1% chlorhexidine gluconate), mitigate intraoperative temperature effects associated with several traditional agents and combinations. Furthermore, reduction of skin bacterial load without adverse effects on healing was seen with fewer than triplicate applications of most tested agents. Ultimately effective skin preparation can be achieved by using only 1 or 2 applications of scrub, thus rendering the triplicate skin-prep method unnecessary in laboratory mice.