Evaluation of Thermal Support during Anesthesia Induction on Body Temperature in C57BL/6 and Nude Mice.

Heat supplementation during surgery is a common practice; however, thermal support is not commonly used during anesthesia induction. Mice lose body temperature quickly, and air movement can exacerbate this, potentially putting mice at a thermal deficit before surgery. Whether the method of warming during induction affects overall heat loss during anesthesia is unknown. We hypothesized that the method of heating would affect body temperature (Tb) during anesthesia induction, maintenance, recovery, and once placed back on the rack. Mice (C57BL/6NHsd-6M/6F [C57BL/6]; Hsd:Athymic Nude-Foxn1nu [Nude]; N = 24;12M/12F) were assigned to a treatment in a factorial design: thermal chamber (TC; ambient temperature [Ta] = 28.8 °C); heating pad (HP; induction chamber placed on an electric heating pad;Ta = 28.4 °C); and control (Ctrl; Ta = 21.6 °C). During induction, one mouse at a time was anesthetized with isoflurane over a 3 min period and then maintained under anesthesia for 10 min on a hot water heating pad (33 °C). Then isoflurane was stopped and time to ambulation was recorded. Tb and activity were tracked in the home cage on the rack before and after anesthesia. During induction, Ctrl mice lost significantly more heat (-2.8 °C) than did TC (+0.2 °C) and HP mice (+0.1 °C) but TC and HP were not different. During anesthesia maintenance, Ctrl mice regained 1 °C, but their Tb was still lower than that of the treated groups. Nude mice consistently had a lower Tb than C57BL/6 mice, regardless of treatment or anesthesia phase. C57BL/6 Ctrl mice took longer to ambulate than either HP or TC mice, but the method of heating did not differentially affect Nude mice. In general, C57BL/6 as compared with Nude and females as compared with males were comparatively more active and had higher Tb during certain times of day, regardless of the heating methods. Overall, our findings support the provision of heat during anesthesia induction, regardless of method, to reduce overall Tb loss during a short anesthesia event.

Recommendations for measuring and standardizing light for laboratory mammals to improve welfare and reproducibility in animal research.

Light enables vision and exerts widespread effects on physiology and behavior, including regulating circadian rhythms, sleep, hormone synthesis, affective state, and cognitive processes. Appropriate lighting in animal facilities may support welfare and ensure that animals enter experiments in an appropriate physiological and behavioral state. Furthermore, proper consideration of light during experimentation is important both when it is explicitly employed as an independent variable and as a general feature of the environment. This Consensus View discusses metrics to use for the quantification of light appropriate for nonhuman mammals and their application to improve animal welfare and the quality of animal research. It provides methods for measuring these metrics, practical guidance for their implementation in husbandry and experimentation, and quantitative guidance on appropriate light exposure for laboratory mammals. The guidance provided has the potential to improve data quality and contribute to reduction and refinement, helping to ensure more ethical animal use.

Opinion: Common Errors, Omissions, and Presentation Problems in the Conduct and Reporting of Animal-based Research.

As experienced authors, statisticians, editors, and scientists, we present the following comments to highlight some usages or omissions that are common in research manuscripts. Consideration of these comments will improve practices of data analysis and reporting.

Evaluating the temperature preferences of sexually mature Duroc, Landrace, and Yorkshire boars.

An accurate understanding of boar temperature preferences may allow the swine industry to design and utilize environmental control systems in boar facilities more precisely. Therefore, the study objective was to determine the temperature preferences of sexually mature Duroc, Landrace, and Yorkshire boars. Eighteen, 8.57 ± 0.10-mo-old boars (N = 6 Duroc, 6 Landrace, and 6 Yorkshire; 186.25 ± 2.25 kg) were individually tested in thermal apparatuses (12.20 m × 1.52 m × 1.86 m) that allowed free choice of their preferred temperature within a 8.92 to 27.92 ºC range. For analyses, the apparatuses were divided into five thermal zones (3.71 m2/thermal zone) with temperature recorded 1.17 m above the floor in the middle of each zone. Target temperatures for thermal zones 1 to 5 were 10, 15, 20, 25, and 30 ºC, respectively. All boars were given a 24-h acclimation phase followed by a 24-h testing phase within the thermal apparatuses. Daily feed allotments (3.63 kg/d) were provided to each boar and all boars were allowed to consume all feed prior to entering the thermal apparatus. Water was provided ad libitum within the thermal apparatuses with 1 waterer per thermal zone. During testing, boars were video recorded continuously to evaluate behavior (inactive, active, or other), posture (lying, standing, or other), and thermal zone the boar occupied. All parameters were recorded in 15 min intervals using instantaneous scan sampling. Data were analyzed using GLM in JMP 15. For the analyses, only time spent lying or inactive were used because they were observed most frequently (lying 80.02%, inactive 77.64%) and were deemed to be associated with comfort based on previous research. Percent time spent active (19.73%) or standing (15.87%) were associated with latrine or drinking activity and were too low to accurately analyze as an indicator of thermal preference. Breed did not affect temperature preference (P > 0.05). A cubic regression model determined that boars spent the majority of their time inactive at 25.50 ºC (P < 0.01) and lying (both sternal and lateral) at 25.90 ºC (P < 0.01). These data suggest that boar thermal preferences did not differ by breed and that boars prefer temperatures at the upper end of current guidelines (10.00 to 25.00 ºC).

Benchmarking Enrichment Efforts in the US & Canada Across Species and Enrichment Categories.

Enrichment is important for animal welfare and data quality. Provision of enrichment opportunities varies between species and enrichment category. However, data benchmarking these differences does not exist. Our objective was to characterize enrichment provision and associated factors across species in the US and Canada. Personnel who work with research animals (n = 1098) in the US and Canada voluntarily responded to online promotions and completed a survey about enrichment used for the species they worked with most, their control of and wish for more enrichment, stress or pain in the animals they worked the most with, and demographics. All participants (except those working with rats) received the same questionnaire regardless of species to allow objectivity, as the effects of many enrichment items on some species have not yet been determined. The questionnaire asked about enrichments that were beneficial to at least one species. The provision of enrichment was allocated into 2 outcome variables: diversity and frequency per enrichment category. Results showed a significant interaction between enrichment category and species. Generally, physical, nutritional, and sensory enrichments were provided less often than social enrichment. In addition, nonhuman primates received more diverse and more frequent enrichment than did other species (twice as much as rats and mice). Enrichment was provided less frequently by personnel who wished they could do more than the status quo. Both enrichment frequency and diversity were higher in respondents from Canada, those who had more control over provision, and those who had been in the field longer. While our results cannot be used to determine the quality of enrichment provided to various species, they do provide information on current enrichment practices in the US and Canada and identify differences in implementation by species and enrichment category. The data also indicate provision of enrichment is influenced by factors such as country and individual control over enrichment. This information can also be used to identify areas for greater enrichment efforts for some species (for example, rats and mice) and categories, with the ultimate goal of improving animal welfare.

A behavior and physiology-based decision support tool to predict thermal comfort and stress in non-pregnant, mid-gestation, and late-gestation sows.

BACKGROUND: Although thermal indices have been proposed for swine, none to our knowledge differentiate by reproductive stage or predict thermal comfort using behavioral and physiological data. The study objective was to develop a behavior and physiology-based decision support tool to predict thermal comfort and stress in multiparous (3.28 ± 0.81) non-pregnant (n = 11), mid-gestation (n = 13), and late-gestation (n = 12) sows. RESULTS: Regression analyses were performed using PROC MIXED in SAS 9.4 to determine the optimal environmental indicator [dry bulb temperature (TDB) and dew point] of heat stress (HS) in non-pregnant, mid-gestation, and late-gestation sows with respiration rate (RR) and body temperature (TB) successively used as the dependent variable in a cubic function. A linear relationship was observed for skin temperature (TS) indicating that TDB rather than the sow HS response impacted TS and so TS was excluded from further analyses. Reproductive stage was significant for all analyses (P < 0.05). Heat stress thresholds for each reproductive stage were calculated using the inflections points of RR for mild HS and TB for moderate and severe HS. Mild HS inflection points differed for non-pregnant, mid-gestation, and late gestation sows and occurred at 25.5, 25.1, and 24.0 °C, respectively. Moderate HS inflection points differed for non-pregnant, mid-gestation, and late gestation sows and occurred at 28.1, 27.8, and 25.5 °C, respectively. Severe HS inflection points were similar for non-pregnant and mid-gestation sows (32.9 °C) but differed for late-gestation sows (30.8 °C). These data were integrated with previously collected behavioral thermal preference data to estimate the TDB that non-pregnant, mid-gestation, and late-gestation sows found to be cool (TDB < TDB preference range), comfortable (TDB = TDB preference range), and warm (TDB preference range < TDB < mild HS). CONCLUSIONS: The results of this study provide valuable information about thermal comfort and thermal stress thresholds in sows at three reproductive stages. The development of a behavior and physiology-based decision support tool to predict thermal comfort and stress in non-pregnant, mid-gestation, and late-gestation sows is expected to provide swine producers with a more accurate means of managing sow environments.

Evaluation of Ambient Sound, Vibration, and Light in Rodent Housing Rooms.

Excessive sound, vibration, and light are detrimental to rodent welfare, yet these parameters are rarely recorded in vivaria. Whether housing environments exceed the suggested thresholds and which specific factors may alter these parameters is generally unknown. The goal of this study was to determine how environmental factors may alter sound, vibration, and light at the room and cage levels. Measurements were made using an ultrasonic microphone, accelerometer, and light sensor. Measurement sites were 1) in open air at a central location in 64 rooms located in 9 buildings, and 2) inside an empty mouse or rat cage containing chow, water, and bedding and located on an animal transfer station (n = 51) or housing rack (n = 102). Information collected for each transfer station and rack measurement included the year of manufacture, the species on the rack, and the number of cages on the rack. For each location, a baseline measurement was taken with the transfer station turned off, followed by another measurement after the transfer station was turned on. In general, many factors influenced ambient sound, vibration, and light, indicating that values are not uniform across rodent rooms in the same institution or across cages in a single room. Sound peaks capable of startling rodents were measured in association with hallway ultrasonic motion sensors and during cage change. Vibration and light intensity were generally low when cages were located on the rack. In contrast, active transfer stations had more vibration and light intensity, reaching levels that were potentially stressful for rodents. These data reflect the ambient sound, vibration, and light that rodents experience during normal facility operations. These patterns may extend to other locations, but given the variability in all parameters, the data highlight the need for institutions to conduct their own monitoring.

Assessing the effect of compounds from plantar foot sweat, nesting material, and urine on social behavior in male mice, Mus musculus.

Home cage aggression causes poor welfare in male laboratory mice and reduces data quality. One of the few proven strategies to reduce aggression involves preserving used nesting material at cage change. Volatile organic compounds from the nesting material and several body fluids not only correlate with less home cage aggression, but with more affiliative allo-grooming behavior. To date, these compounds have not been tested for a direct influence on male mouse social behavior. This study aimed to determine if 4 previously identified volatile compounds impact home cage interactions. A factorial design was used with cages equally split between C57BL/6N and SJL male mice (N = 40). Treatments were randomly assigned across cages and administered by spraying one compound solution on each cage's nesting material. Treatments were refreshed after day 3 and during cage change on day 7. Home cage social behavior was observed throughout the study week and immediately after cage change. Several hours after cage change, feces were collected from individual mice to measure corticosterone metabolites as an index of social stress. Wound severity was also assessed after euthanasia. Measures were analyzed with mixed models. Compound treatments did not impact most study measures. For behavior, SJL mice performed more aggression and submission, and C57BL/6N mice performed more allo-grooming. Wound severity was highest in the posterior region of both strains, and the middle back region of C57BL/6N mice. Posterior wounding also increased with more observed aggression. Corticosterone metabolites were higher in C57BL/6N mice and in mice treated with 3,4-dimethyl-1,2-cyclopentanedione with more wounding. These data confirm previous strain patterns in social behavior and further validates wound assessment as a measure of escalated aggression. The lack of observed treatment effects could be due to limitations in the compound administration procedure and/or the previous correlation study, which is further discussed.

Protein profiles from used nesting material, saliva, and urine correspond with social behavior in group housed male mice, Mus musculus.

Current understanding of how odors impact intra-sex social behavior is based on those that increase intermale aggression. Yet, odors are often promoted to reduce fighting among male laboratory mice. It has been shown that a cage of male mice contains many proteins used for identification purposes. However, it is unknown if these proteins relate to social behavior or if they are uniformly produced across strains. This study aimed to compare proteomes from used nesting material and three sources (sweat, saliva, and urine) from three strains and compare levels of known protein odors with rates of social behavior. Used nesting material samples from each cage were analyzed using LC-MS/MS. Sweat, saliva, and urine samples from each cage's dominant and subordinate mouse were also analyzed. Proteomes were assessed using principal component analyses and compared to behavior by calculating correlation coefficients between PC scores and behavior proportions. Twenty-one proteins from nesting material either correlated with affiliative behavior or negatively correlated with aggression. Notably, proteins from the major urinary protein family, odorant binding protein family, and secretoglobin family displayed at least one of these patterns, making them candidates for future work. These findings provide preliminary information about how proteins can influence male mouse behavior. SIGNIFICANCE: Research on how olfactory signals influence same sex social behavior is primarily limited to those that promote intermale aggression. However, exploring how olfaction modulates a more diverse behavioral repertoire will improve our foundational understanding of this sensory modality. In this proteome analysis we identified a short list of protein signals that correspond to lower rates of aggression and higher rates of socio-positive behavior. While this study is only correlational, it sets a foundation for future work that can identify protein signals that directly influence social behavior and potentially identify new murine pheromones.

Home cage measures of Alzheimer's disease in the rTg4510 mouse model.

Alzheimer's disease affects an array of activities in patients' daily lives but measures other than memory are rarely evaluated in animal models. Home cage behavior, however, may provide an opportunity to back translate a variety of measures seen in human disease progression to animal models, providing external and face validity. The aim of this study was to evaluate if home cage measures could indicate disease in the rTg4510 mouse model. We hypothesized that sleep, nesting, and smell discrimination would be altered in mutant mice. Thirty-two transgenic mice were used in a Latin square design of four genotypes x both sexes x two diets. Half the mice received a doxycycline diet to suppress tauopathy and evaluate tau severity on various measures. At 8-, 12-, and 16-weeks old, 24 h activity/sleep patterns, nest complexity, and odor discrimination were measured. After 16-weeks, tau concentration in the brain was quantified. Mutant mice had increased tau concentration in brain tissue, but it was reduced by the doxycycline diet. However, only nest complexity was different between mutant mice and controls. Overall, tauopathy in rTg4510 mice does seem to affect these commonly observed symptoms in human patients. However, while running this study, a report showed that the rTg4510 mutant phenotype is not caused by the mutation itself, but confounding factors from transgene insertion. Combined with report findings and our data, the rTg4510 model may not be an ideal model for all aspects of human Alzheimer's disease.

Characterizing the effect of incrementally increasing dry bulb temperature on linear and nonlinear measures of heart rate variability in nonpregnant, mid-gestation, and late-gestation sows.

Characterizing the sow physiological response to an increased heat load is essential for effective heat stress mitigation. The study objective was to characterize the effects of a 400-min heating episode on sow heart rate variability (HRV) at different reproductive stages. HRV is a commonly used noninvasive proxy measure of autonomic function. Twenty-seven sows were enrolled in the study according to their gestation stage at time of selection: 1) nonpregnant (NP; n = 7), 2) mid-gestation (MID; 57.3 ± 11.8 d gestation; n = 11), and 3) late-gestation (LATE; 98.8 ± 4.9 d gestation; n = 8). The HRV data utilized in the study were collected from each pig as the dry bulb temperature in the room increased incrementally from 19.84 ± 2.15 °C to 35.54 ± 0.43 °C (range: 17.1-37.5 °C) over a 400-min period. After data collection, one 5-min set of continuous heart rate data were identified per pig for each of nine temperature intervals (19-20.99, 21-22.99, 23-24.99, 25-26.99, 27-28.99, 29-30.99, 31-32.99, 33-34.99, and 35-36.99 °C). Mean inter-beat interval length (RR), standard deviation of r-r intervals (SDNN), root mean square of successive differences (RMSSD), high frequency spectral power (HF), sample entropy (SampEn), short-term detrended fluctuation analysis (DFAα1), and three measures (%REC, DET, LMEAN) derived from recurrence quantification analysis were calculated for each data set. All data were analyzed using the PROC GLIMMIX procedure in SAS 9.4. Overall, LATE sows exhibited lower RR than NP sows (P < 0.01). The standard deviation of r-r intervals and RMSSD differed between each group (P < 0.01), with LATE sows exhibiting the lowest SDNN and RMSSD and NP sows exhibiting the greatest SDNN and RMSSD. Late-gestation sows exhibited lower HF than both MID and NP sows (P < 0.0001), greater DFA values than NP sows (P = 0.05), and greater DET compared to MID sows (P = 0.001). Late-gestation also sows exhibited greater %REC and LMEAN compared to MID (P < 0.01) and NP sows (all P < 0.01). In conclusion, LATE sows exhibited indicators of greater autonomic stress throughout the heating period compared to MID and NP sows. However, temperature by treatment interactions were not detected as dry bulb increased. Future studies are needed to fully elucidate the effect of gestational stage and increasing dry bulb temperature on sow HRV.

Pregnant pigs may be at a higher risk of poor physiological outcomes due to heat exposure compared to mature female pigs that are not pregnant. The purpose of this study was to evaluate the stress response of pregnant pigs to increasing environmental temperatures using heart rate variability, a noninvasive measure commonly used to evaluate the physiological stress response. Our findings show that pregnant pigs, particularly those who are closer to giving birth, exhibit greater evidence of physiological stress compared to pigs who are not pregnant. However, we did not find evidence that increasing environmental temperature throughout the experimental period was a primary reason for the increased stress exhibited by pregnant pigs. It is possible that the physiological changes that normally occur during pregnancy may have masked the physiological stress response typically associated with increased heat exposure.

Evaluation of sow thermal preference across three stages of reproduction.

The metabolic heat production of modern pigs has increased by an average of 16%, compared with sows of 30 years ago. Therefore, it is likely that temperature recommendations require updating to meet the needs of modern pigs. The objective of this study was to evaluate whether different reproductive stages of sows altered thermal preference and if current recommendations required updating. Twenty multiparous sows (3.4 ± 1.2 parity) in different reproductive stages (nonpregnant: n = 7; mid-gestation: 58.5 ± 5.68 d, n = 6; and late-gestation: 104.7 ± 2.8 d, n = 7) were tested. Thermal preference was individually tested, and sows could freely choose a temperature, using a thermal gradient between 10.4 and 30.5 °C. Sows were given 24 h to acclimate to the thermal apparatus. Before testing began, sows were given daily feed allotment and returned to the apparatus. Video from the 24-h test period was used to record sow behavior (time spent inactive), posture (upright and sternal and lateral lying), and location using instantaneous scan samples every 15 min. Data were analyzed using PROC MIXED procedure in SAS 9.4. A cubic regression model was used to calculate the sow's most preferred temperature based on the location, or temperature, in which they spent the most time. The preference range was calculated using peak temperature preference ±SE for each sow. The reproductive stage altered where sows spent their time within the thermal gradient (P < 0.01). Late-gestation sows preferred cooler temperatures (14.0 °C) than mid-gestation (14.8 °C; P < 0.01) and nonpregnant sows (14.8 °C; P < 0.01). In summary, sow thermal preferences were within the lower half of the current recommended range (10 to 25 °C). This indicates that temperatures at the higher end of the recommended range could be uncomfortable to sows and that the thermal comfort zone of sows may be narrower than recommendations indicate.

Who's the Boss? Assessing Convergent Validity of Aggression Based Dominance Measures in Male Laboratory Mice, Mus Musculus.

Aggression among group housed male mice continues to challenge laboratory animal researchers because mitigation strategies are generally applied at the cage level without a good understanding of how it affects the dominance hierarchy. Aggression within a group is typically displayed by the dominant mouse targeting lower ranking subordinates; thus, the strategies for preventing aggression may be more successful if applied specifically to the dominant mouse. Unfortunately, dominance rank is often not assessed because of time intensive observations or tests. Several correlates of dominance status have been identified, but none have been directly compared to home cage behavior in standard housing. This study assessed the convergent validity of three dominance correlates (urinary darcin, tube test score, preputial gland to body length ratio) with wound severity and rankings based on home cage behavior, using factor analysis. Discriminant validity with open field measures was assessed to determine if tube test scores are independent of anxiety. Cages were equally split between SJL and albino C57BL/6 strains and group sizes of 3 or 5 (N = 24). Home cage behavior was observed during the first week, and dominance measures were recorded over the second. After controlling for strain and group size, darcin and preputial ratio had strong loadings on the same factor, which was a significant predictor of home cage ranking showing strong convergent validity. Tube test scores were not significantly impacted by open field data, showing discriminant validity. Social network analysis revealed that despotic power structures were prevalent, aggressors were typically more active and rested away from cage mates, and the amount of social investigation and aggression performed by an individual were highly correlated. Data from this study show that darcin and preputial ratio are representative of home cage aggression and provide further insight into individual behavior patterns in group housed male mice.

Thermoregulatory and physiological responses of nonpregnant, mid-gestation, and late-gestation sows exposed to incrementally increasing dry bulb temperature.

Gestating sows may be more susceptible to increasing dry bulb temperatures (TDB) due to greater metabolic heat production and increased body mass, especially as gestation advances. However, there are few studies on the thermoregulatory and physiological responses of sows at differing gestation stages exposed to gradually increasing temperatures. The study objective was to determine the thermoregulatory and physiological responses of nonpregnant (n = 12; parity 3.27 ± 0.86), mid-gestation (59.7 ± 9.6 d pregnant, n = 12; parity 3.25 ± 0.83), and late-gestation (99.0 ± 4.8 d pregnant, n = 12; parity 3.33 ± 0.75) sows exposed to increasing TDB. Prior to the experiment (5.0 ± 0.7 d), jugular catheters were placed in all sows. During the experiment, the TDB was increased incrementally by 2.45 ± 0.43 °C every 60 min from 19.84 ± 2.15 to 35.54 ± 0.43 °C over 400 min, and relative humidity was recorded at 40.49 ± 18.57%. Respiration rate (RR), heart rate (HR), skin temperature, and vaginal temperature were measured, and blood samples were obtained via the jugular catheter every 20 min. Data were analyzed using PROC MIXED in SAS 9.4. RR increased at a lower TDB (P < 0.01) in late-gestation sows compared with mid-gestation and nonpregnant sows, but no differences were detected between mid-gestation and nonpregnant sows. Overall, late-gestation sows had greater RR (P < 0.01; 23 ± 2 breaths per min [brpm]) compared with mid-gestation (16 ± 2 brpm) and nonpregnant (15 ± 2 brpm) sows. Late-gestation sows had an overall greater HR (P < 0.01; 84 ± 5 beats per min [bpm]) than mid-gestation (76 ± 5 bpm) and nonpregnant (69 ± 5 bpm) sows. Late-gestation sows had overall reduced bicarbonate and total carbon dioxide levels (P = 0.02; 23.89 ± 1.97 and 25.41 ± 2.07 mmol/L, respectively) compared with mid-gestation (27.03 ± 1.97 and 28.58 ± 2.07 mmol/L, respectively) and nonpregnant (26.08 ± 1.97 and 27.58 ± 2.07 mmol/L, respectively) sows. Moreover, late-gestation sows had overall greater nitric oxide levels (P < 0.01; 248.82 ± 34.54 µM) compared with mid-gestation (110.47 ± 34.54 µM) and nonpregnant (41.55 ± 34.54 µM) sows. In summary, late-gestation sows appear to be more sensitive to increasing TDB as indicated by thermoregulatory and physiological responses when compared with mid-gestation or nonpregnant sows. The results from this study provide valuable information regarding thermoregulatory thresholds of sows at differing gestation stages.

One Is the Coldest Number: How Group Size and Body Weight Affect Thermal Preference in Weaned Pigs (3 to 15 kg).

Housing pigs within their thermal comfort zone positively impacts productivity and performance. However, fundamental information on behavioral thermoregulatory responses of individual and group-housed pigs is meager. As a gregarious species, pigs prefer to be near one another, touching and often huddling. As pigs huddle together, they decrease their heat loss to the environment by decreasing exposed surface area and increasing mass. Additionally, pigs gain weight rapidly as they age. As an individual grows, their ability to withstand lower temperatures increases. We hypothesized that group size would alter pig thermal preference and that thermal preference would change based upon body weight. Thirty-six groups of pigs (n = 2 pigs/group) were tested in a factorial design based on group size (1, 2, or 4) and weight category (small: 5.20 ± 1.15 kg; medium: 8.79 ± 1.30 kg; and large: 13.95 ± 1.26 kg) in both sexes. Treatment groups were placed inside a chamber with a controlled thermal gradient (4.6 m × 0.9 m × 0.9 m; L × W × H) that ranged in temperature from 18 to 30 °C. Pigs habituated to the gradient for 24 h. The following 24 h testing period was continuously video recorded and each pig's location during inactivity (~70% daily budget) within the thermal apparatus was recorded every 10 min via instantaneous scan sampling. Data were analyzed using a GLM and log10 + 0.001 transformed for normality. Tukey tests and Bonferroni-corrected custom tests were used for post hoc comparisons. Peak temperature preference was determined by the maximum amount of time spent at a specific temperature. Both group size (p = 0.001) and weight category (p < 0.001) influenced the thermal location choice of pigs. Individual pigs preferred 30.31 °C, which differed from a group of 2 (20.0 °C: p = 0.003) and 4 pigs (20.0 °C: p < 0.001). The peak temperature preference of the small pigs (30.2 °C) differed from the large pigs (20.0 °C: p < 0.001) but did not differ from the medium-sized pigs (28.4 °C: p > 0.05). Overall, heavier pigs and larger groups preferred cooler temperatures.

Compounds from plantar foot sweat, nesting material, and urine show strain patterns associated with agonistic and affiliative behaviors in group housed male mice, Mus musculus.

Excessive home cage aggression often results in severe injury and subsequent premature euthanasia of male laboratory mice. Aggression can be reduced by transferring used nesting material during cage cleaning, which is thought to contain aggression appeasing odors from the plantar sweat glands. However, neither the composition of plantar sweat nor the deposits on used nesting material have been evaluated. The aims of this study were to (1) identify and quantify volatile compounds deposited in the nest site and (2) determine if nest and sweat compounds correlate with social behavior. Home cage aggression and affiliative behavior were evaluated in 3 strains: SJL, C57BL/6N, and A/J. Individual social rank was assessed via the tube test, because ranking may influence compound levels. Sweat and urine from the dominant and subordinate mouse in each cage, plus cage level nest samples were analyzed for volatile compound content using gas chromatography-mass spectrometry. Behavior data and odors from the nest, sweat, and urine were statistically analyzed with separate principal component analyses (PCA). Significant components, from each sample analysis, and strain were run in mixed models to test if odors were associated with behavior. Aggressive and affiliative behaviors were primarily impacted by strain. However, compound PCs were also impacted by strain, showing that strain accounts for any relationship between odors and behavior. C57BL/6N cages displayed the most allo-grooming behavior and had high scores on sweat PC1. SJL cages displayed the most aggression, with high scores on urine PC2 and low scores on nest PC1. These data show that certain compounds in nesting material, urine, and sweat display strain specific patterns which match strain specific behavior patterns. These results provide preliminary information about the connection between home cage compounds and behavior. Salient compounds will be candidates for future controlled studies to determine their direct effect on mouse social behavior.

Identification of Sick or Dead Mice (Mus musculus) Housed with 6 Grams of Crinkle Paper Nesting Material.

Although nesting material is beneficial to the welfare of laboratory mice, provision of appropriate amounts may impair visualization of the mice. In anticipation of our academic research institution transitioning to providing 6 grams of nesting material to all mice, we conducted a 2-step prospective epidemiologic study to 1) evaluate whether 0, 2, or 6 grams of nesting material alters the ability to identify sick or dead mice, and 2) evaluate the number and severity of health concerns identified in the presence of 6 grams of crinkle paper nesting material at cage-side health check as compared with cage change. Animal Treatment Reports (ATRs) and death incidences were collected across a variety of research and breeding uses. This information was used to determine if nesting material prevented prompt identification of mice in need of veterinary attention. The clinical health condition category (CHCC) was determined based on the severity of the animal's health condition on initial veterinary exam. Additional assessment determined if the identification of the animal's condition was a success (early-stage or mild illness when first identified) or a failure (late-stage or endstage illness when first identified). Mice that died spontaneously were also assessed with regard to which observation activity was being performed at the time of the animal's identification (daily health check or cage change) and location of the mouse in relation to the nest. The results showed that nesting material did not cause a significant increase in the severity of CHCCs at the time reported for veterinary evaluation. Successful identification of health concerns occurred significantly more often than failures. Death rates were similar between all nesting groups, and dead mice were more likely to be located outside of the nest. In summary, nesting material did not hinder the ability to identify mice in need of veterinary care during routine cage-side health checks and did not critically affect the ability to identify mice that died spontaneously. These results indicate that mice can receive appropriate amounts of crinkle paper nesting material without lowering the ability of staff to recognize mice in need of veterinary attention.

Early life thermal stress: impacts on future temperature preference in weaned pigs (3 to 15 kg).

Thermal stress can result in productivity losses, morbidity, and mortality if proper management practices are not employed. A basic understanding of the relationship between animals and the thermal environment is crucial to assess the environment's impact on livestock performance. Therefore, the study objective was to evaluate whether different early life thermal stressors (ELTS) altered the temperature preference of pigs later in life. Twelve sows and their litters were randomly exposed to 1 of 3 ELTS treatments from 7 to 9 d of age: early life heat stress (ELHS; cycling 32 to 38 °C; n = 4), early life cold stress (ELCS; 25.4±1.1 °C without heating lamp; n = 4), or early life thermoneutral (ELTN; 25.4±1.1 °C with a heating lamp; n = 4) conditions. From 10 to 20 d, (weaning) all piglets were exposed to ELTN conditions. At weaning, pigs were randomly assigned to groups of 4 of the same sex and ELTS treatment. Temperature preference, where pigs freely choose a temperature, was assessed in 21 groups (n = 7 groups per ELTS treatment) using 1 of 3 thermal gradient apparatuses (22 to 40 °C). Testing began at 26 ± 1.3 d of age to give pigs time to acclimate to solid food after weaning and 1 group per ELTS treatment were tested simultaneously in each apparatus. Pigs were given 24 h to acclimate followed by a 24-h testing period. Behavior (active and inactive), posture (upright, sternal, and lateral lying), and location were documented every 20 min using instantaneous scan samples. Preferred feeding temperature was determined by the latency to empty a feeder in each location. Data were analyzed using PROC MIXED in SAS 9.4. A cubic regression model was used to calculate the peak temperature preference of pigs based on the temperature pigs spent most of their time. The preference range was calculated using peak temperature preference ±SE for each ELTS treatment group. Early life thermal stress altered where pigs spent most of their time within the thermal gradient (P = 0.03) with ELTN pigs preferring cooler temperatures (peak preference of 23.8 °C) compared with their ELCS exposed counterparts (peak preference of 26.0 °C; P < 0.01). However, ELHS exposed pigs (peak preference of 25.6 °C) did not differ in their temperature preference compared with ELTN or ELCS exposed counterparts (P > 0.05). In summary, ELCS exposure altered pig temperature preference later in life indicating that ELTS can alter temperature preference in pigs.

The epidemiology of fighting in group-housed laboratory mice.

Injurious home-cage aggression (fighting) in mice affects both animal welfare and scientific validity. It is arguably the most common potentially preventable morbidity in mouse facilities. Existing literature on mouse aggression almost exclusively examines territorial aggression induced by introducing a stimulus mouse into the home-cage of a singly housed mouse (i.e. the resident/intruder test). However, fighting occurring in mice living together in long-term groups under standard laboratory housing conditions has barely been studied. We performed a point-prevalence epidemiological survey of fighting at a research institution with an approximate 60,000 cage census. A subset of cages was sampled over the course of a year and factors potentially influencing home-cage fighting were recorded. Fighting was almost exclusively seen in group-housed male mice. Approximately 14% of group-housed male cages were observed with fighting animals in brief behavioral observations, but only 14% of those cages with fighting had skin injuries observable from cage-side. Thus simple cage-side checks may be missing the majority of fighting mice. Housing system (the combination of cage ventilation and bedding type), genetic background, time of year, cage location on the rack, and rack orientation in the room were significant risk factors predicting fighting. Of these predictors, only bedding type is easily manipulated to mitigate fighting. Cage ventilation and rack orientation often cannot be changed in modern vivaria, as they are baked in by cookie-cutter architectural approaches to facility design. This study emphasizes the need to invest in assessing the welfare costs of new housing and husbandry systems before implementing them.

Changing Human Behavior to Improve Animal Welfare: A Longitudinal Investigation of Training Laboratory Animal Personnel about Heterospecific Play or "Rat Tickling".

Despite evidence for rat tickling's animal welfare benefits, the technique is rarely implemented in part because of a lack of training. This study's purpose was to determine the efficacy of online-only or online + hands-on training programs on key outcomes for rat tickling in comparison to a waitlist control condition. After completing a baseline survey, laboratory animal personnel currently working with rats in the United States were semi-randomized to receive online-only training (n = 30), online + hands-on training (n = 34), or waitlist control (n = 32). Participants received further surveys directly after training and 2 months later. Data were analyzed using general linear mixed models. At the 2-month follow-up compared to baseline, both training groups reported increased implementation, self-efficacy, knowledge, and familiarity of rat tickling while only the online + hands-on training participants reported increased control beliefs (while the waitlist group stayed the same). At the 2-month follow-up compared to the waitlist, hands-on training participants reported increased self-efficacy and familiarity with rat tickling. Overall, findings show that both online-only and online + hands-on training can improve key outcomes for rat tickling. Although online + hands-on training is slightly more effective, the interactive online-only training has the potential to improve widescale implementation of a welfare-enhancing technique.