Transient hypertension and sustained tachycardia in mice housed individually in metabolism cages

The novel environment of a metabolic cage can be stressful for rodents, but few studies have attempted to quantify this stress-response. Therefore, we determined the effects on mean arterial pressure (MAP) and heart rate (HR), of placing mice of both sexes in metabolism cages for 2 days. After surgical implantation of a carotid artery catheter mice recovered individually in standard cages for 5 days. Mice then spent 2 days in metabolism cages. MAP and HR were monitored in the standard cage on Day 5 and in metabolism cages on Days 6-7. MAP increased by 18+/-3 and 22+/-4 %, while HR increased by 27+/-4 and 27+/-6 %, in males and females, respectively, during the first hours after cage switch. MAP decreased to baseline in the fourth and eighth h following metabolism cage switch in males and females, respectively. However, HR remained significantly elevated in both sexes during the entire two-day period in metabolism cages. Females had lower MAP than males both pre- and post-metabolism cage switch, but there were no sex differences in HR. These results demonstrate sustained changes in cardiovascular function when mice are housed in metabolism cages, which could potentially affect renal function.     

Effects of housing density on nasal pathology of breeding mice housed in individually ventilated cages

The 2011 edition of the Guide for the Care and Use of Laboratory Animals includes new recommendations for the amount of floor space that should be provided to breeding mice. When pairs or trios of continuously breeding mice are housed in shoebox cages, they may have less than this recommended amount of floor space. High housing densities may adversely affect animal health, for example, by compromising air quality inside the cage. Hence, some institutions are carefully reevaluating the microenvironments of breeding cages. The use of individually ventilated cages (IVCs) to house research mice allows for greater control over the quality of the cage microenvironment. The authors evaluated the microenvironments of shoebox cages in an IVC rack system housing breeding and non-breeding Swiss Webster mice. Ammonia concentrations were significantly higher in cages housing breeding trios with two litters. Histopathologic lesions attributable to inhaled irritants such as ammonia were found in mice housed in breeding pairs and trios. The authors conclude that the microenvironments of cages in an IVC rack system housing breeding pairs and trios may be detrimental to animal health.     

Influence of environmental enrichment and handling on the acute stress response in individually housed mice

In this study we investigated the effect of environmental enrichment and handling on the acute physiological stress response caused by short periods of restraint in individually housed female mice. Heart rate (HR) and body temperature (BT) were measured by radiotelemetry and compared with plasma corticosterone (pCORT) levels. Also, postmortem thymus weight and tyrosine hydroxylase (TH) activity were assessed. The acute stress response was seen in both HR and BT. Enrichment and handling were found to increase rather than decrease this stress response, but pCORT values, measured 90 min after restraint, suggested a lower stress response in the enriched groups. No effect was found with thymus weight or TH as parameters.     

Cage toys reduce abnormal behavior in individually housed pigtail macaques

As part of a behavioral intervention program that identifies and treats individual nonhuman primates exhibiting abnormal behavior, five individually housed pigtail macaques (Macaca nemestrina) were provided with multiple cage toys in an effort to reduce high levels of abnormal behavior. Ten 30-min observations of each subject were conducted during the baseline condition and again after novel toys were presented, both loose inside the cage and attached to the outside of the cage. The new toys were used during 27% of the observation time. Kong Toys were used most consistently by the macaques during the 5-week observation period. Significant decreases in abnormal behavior and cage-directed behavior, as well as significantly increased enrichment use, were evident after the toys were added. Several of the toys were destroyed quickly, and individual differences were evident in the levels of enrichment use and abnormal behavior. Providing multiple manipulable toys as enrichment for pigtail macaques was effective in reducing abnormal behavior and was an important part of an environmental enrichment program for monkeys who could not be housed socially.     

Effect of castration on rabbits housed in littermate pairs

Conventional solitary caging for research rabbits precludes many postural and social behaviors. Several studies that have investigated enrichment strategies for rabbit does have found intact males usually too aggressive for paired or group housing. This study investigated the possibilities that male siblings will remain compatible if never separated from birth and that castration will improve compatibility among young sibling male rabbits. As a side benefit of pair housing for all rabbits, the larger cages (modified dog cages) allowed rest and other postures not possible in conventional single cages. In addition, there was no evidence of the stereotypies observed among individually housed rabbits. Castrated, post-weanling, male, Flemish Giant x Giant Chinchilla rabbit littermates demonstrated a significantly higher level of living together compatibly than did the intact controls. All rabbits chose to rest in close proximity or contact, although this preference was stronger in castrated males. During puberty, intact males displayed more serious aggression. We conclude that young adult littermate rabbits can live compatibly as pairs and that castration improves compatibility among males. Paired rabbits exhibit many postures and behaviors that are not possible in the small, solitary cages currently listed as minimum recommendations.     

A programmable environmental system for maintenance and observation of individually housed ticks

This paper describes an inexpensive, space-saving, and labor-efficient method for housing and making repeated observations of individual ticks held under specific photoperiodic, temperature, and humidity conditions. Using 96-well assay plates to house ticks, various developmental parameters, sex, survival, etc., of several thousand individual ticks can be recorded routinely in less than 1 hr. This design also permits more than 3,500 individual ticks to be maintained in 1 small (18.9-L) aquarium that serves as a humidity chamber. A small, unilluminated constant temperature incubator was converted inexpensively into a light and temperature programmable unit to contain the aquarium by adding a timing system, light fixture, and a second thermostat.     

Population density and growth rate in laboratory mice

Home Office guidelines recommend an area of 60 cm2 per mouse for growing mice up to 30 g. However, the overall growth rate, and final adrenal weight of weanling BALB/c and MF1 strain mice was not affected by being housed at a density of down to 27 cm2 per mouse, though there was some evidence of strain differences in ability to tolerate such dense housing. The presence of cage accessories had no effect on growth rate of BALB/c and female mice, but reduced growth of MF1 and male mice, though the effect was small. It is concluded that present Home Office guidelines make a generous provision for the space requirements of growing laboratory mice, and that the use of cage accessories of varying design may be worth exploring in more detail.     

Microbiological monitoring of laboratory mice and biocontainment in individually ventilated cages: a field study

Over recent years, the use of individually ventilated cage (IVC) rack systems in laboratory rodent facilities has increased. Since every cage in an IVC rack may be assumed to be a separate microbiological unit, comprehensive microbiological monitoring of animals kept in IVCs has become a challenging task, which may be addressed by the appropriate use of sentinel mice. Traditionally, these sentinels have been exposed to soiled bedding but more recently, the concept of exposure to exhaust air has been considered. The work reported here was aimed firstly at testing the efficiency of a sentinel-based microbiological monitoring programme under field conditions in a quarantine unit and in a multi-user unit with frequent imports of mouse colonies from various sources. Secondly, it was aimed at determining biocontainment of naturally infected mice kept in an IVC rack, which included breeding of the mice. Sentinels were exposed both to soiled bedding and to exhaust air. The mice which were used in the study carried prevalent infectious agents encountered in research animal facilities including mouse hepatitis virus (MHV), mouse parvovirus (MPV), intestinal flagellates and pinworms. Our data indicate that the sentinel-based health monitoring programme allowed rapid detection of MHV, intestinal flagellates and pinworms investigated by a combination of soiled bedding and exhaust air exposure. MHV was also detected by exposure to exhaust air only. The IVC rack used in this study provided biocontainment when infected mice were kept together with non-infected mice in separate cages in the same IVC rack.     

The genetic contribution to heart rate and heart rate variability in quiescent mice

Recent studies have suggested a genetic component to heart rate (HR) and HR variability (HRV). However, a systematic examination of the genetic contribution to the variation in HR and HRV has not been performed. This study investigated the genetic contribution to HR and HRV using a wide range of inbred and recombinant inbred (RI) mouse strains. Electrocardiogram data were recorded from 30 strains of inbred mice and 29 RI strains. Significant differences in mean HR and total power (TP) HRV were identified between inbred strains and RI strains. Multiple significant differences within the strain sets in mean low-frequency (LF) and high-frequency (HF) power were also found. No statistically significant concordance was found between strain distribution patterns for HR and HRV phenotypes. Genomewide interval mapping identified a significant quantitative trait locus (QTL) for HR [LOD (likelihood of the odds) score = 3.763] on chromosome 6 [peak at 53.69 megabases (Mb); designated HR 1 (Hr1)]. Suggestive QTLs for TP were found on chromosomes 2, 4, 5, 6, and 14. A suggestive QTL for LF was found on chromosome 16; for HF, we found one significant QTL on chromosome 5 (LOD score = 3.107) [peak at 53.56 Mb; designated HRV-high-frequency 1 (Hrvhf1)] and three suggestive QTLs on chromosomes 2, 11 and 15. In conclusion, the results demonstrate a strong genetic component in the regulation of resting HR and HRV evidenced by the significant differences between strains. A lack of correlation between HR and HRV phenotypes in some inbred strains suggests that different sets of genes control the phenotypes. Furthermore, QTLs were found that will provide important insight to the genetic regulation of HR and HRV at rest.     

Nonshivering thermogenesis capacity versus basal metabolic rate in laboratory mice

I wanted to follow the correlation between level of basal metabolic rate (BMR) and maximum response to injection of noradrenaline (MMR_NA) in two lines of laboratory mice subjected to divergent, artificial selection toward high BMR (HBMR) and low BMR (LBMR). HBMR animals had heavier visceral organs (heart, liver, kidney, intestine), but their regulatory NST (MMR_NA–BMR) was lower and interscapular brown adipose tissue (IBAT) lighter than in LBMR mice. Obligatory part of nonshivering thermogenesis (NST) (in other words BMR) depended on visceral organ mass, whereas regulatory NST correlates with mass of IBAT. BMR was not correlated with total NST capacity, but phenotypic correlation between obligatory and regulatory NST was negative. This suggests possibility of substitution of obligatory NST to thermoregulation in a place of the regulatory NST. Then total thermoregulatory energy expenditures do not change.     

Sympathetic control of heart rate in nNOS knockout mice

Inhibition of neuronal nitric oxide synthase (nNOS) in cardiac postganglionic sympathetic neurons leads to enhanced cardiac sympathetic responsiveness in normal animals, as well as in animal models of cardiovascular diseases. We used isolated atria from mice with selective genetic disruption of nNOS (nNOS(-/-)) and their wild-type littermates (WT) to investigate whether sympathetic heart rate (HR) responses were dependent on nNOS. Immunohistochemistry was initially used to determine the presence of nNOS in sympathetic [tyrosine hydroxylase (TH) immunoreactive] nerve terminals in the mouse sinoatrial node (SAN). After this, the effects of postganglionic sympathetic nerve stimulation (1-10 Hz) and bath-applied norepinephrine (NE; 10(-8)-10(-4) mol/l) on HR were examined in atria from nNOS(-/-) and WT mice. In the SAN region of WT mice, TH and nNOS immunoreactivity was virtually never colocalized in nerve fibers. nNOS(-/-) atria showed significantly reduced HR responses to sympathetic nerve activation and NE (P < 0.05). Similarly, the positive chronotropic response to the adenylate cyclase activator forskolin (10(-7)-10(-5) mol/l) was attenuated in nNOS(-/-) atria (P < 0.05). Constitutive NOS inhibition with L-nitroarginine (0.1 mmol/l) did not affect the sympathetic HR responses in nNOS(-/-) and WT atria. The paucity of nNOS in the sympathetic innervation of the mouse SAN, in addition to the attenuated HR responses to neuronal and applied NE, indicates that presynaptic sympathetic neuronal NO does not modulate neuronal NE release and SAN pacemaking in this species. It appears that genetic deletion of nNOS results in the inhibition of adrenergic-adenylate cyclase signaling within SAN myocytes.     

Activity patterns of gray wolves housed in small vs. large enclosures

Free-ranging gray wolves (Canis lupus) generally inhabit large home ranges, yet they are housed in a variety of restricted spaces when in captivity. There is continual debate as to whether space restrictions alter a wolf's behavior. The purpose of these studies was to remotely measure and then compare the amount and frequency of activity of gray wolves housed in small, artificial enclosures vs. large, more natural enclosures. Test animals comprised three adult wolves housed in kennels and three and four wolves housed in separate natural enclosures. Kenneled wolves had 2.8 m² of surface area per wolf, and wolves in natural enclosures had 466.6 m² (South Pack) and 349.9 m² (North Pack) per wolf. Wolves were fitted with radiotelemetry collars containing activity sensors. Activity data were recorded every 20 min for 57 continuous hr. The amount of activity for each wolf was calculated using areas under the curve (AUCs), and the frequency of activity was analyzed by spectral analysis. There was no difference (P ≥ 0.22) in AUCs between kenneled wolves (1.399 ± 0.214 x 10⁵ radians) and South Pack wolves (1.564 ± 0.139 X 10⁵ radians) or North Pack wolves (1.617 ± 0.192 x 10⁵ radians). All three groups had similar peak spectral values at frequencies that were close to daily cycles (i.e., ω = 0.12–0.17 cycles per unit time). Peaks in coherence near the dominant spectral frequency were most significant between the natural enclosures and the least significant between the kenneled wolves and the South Pack wolves. Based on these criteria of activity and under these circumstances, enclosure size appeared to have no effect on wolf activity. However, small sample sizes and variation in the data do not make these results definitive. © 1996 Wiley-Liss, Inc.     

The effect of group housing on feeding patterns and social behaviour of previously individually housed growing pigs

Group housed pigs make less frequent feeder visits of longer duration, and eat at a faster rate than pigs housed individually. They also have lower growth rates which may be due to elevated stress levels resulting from changes in the concentrations of hormones, such as cortisol and adrenaline associated with aggression and social stress. The aim of this experiment was to investigate the effects of grouping on feeding pattern, time budgeting and the social behaviour of pigs kept as individuals from weaning until grouping. In total, 12 Large WhitexLandrace male pigs (four pigs per block) mean (+/-S.E.) start weight 22.5+/-0.7kg were housed individually for 3 weeks (Period 1) after which in two replicates (Blocks 1 and 3), pigs were combined into a group of four (Period 2) before being returned to individual housing for a further 3 weeks (Period 3). In Block 2, the four pigs remained as individuals across periods but were moved between pens at the end of Periods 1 and 2 to account for any pen effects. Feeding pattern and food intake were recorded throughout and pigs were weighed three times a week. Video recordings and live behavioural observations were made to record time budgets and social behaviour. Grouped pigs made less visits to the feeder in Period 2 than when they were housed individually in Periods 1 (P<0.001) and 3 (P<0.01). Visit duration was longer in Period 2 than in Periods 1 (P<0.01) and 3 (P<0.05). Food intake and weight gain were greater in Period 3 than in Periods 1 and 2 (P<0.001 and P<0.01, respectively). There were no significant effects of moving pigs between pens in Block 2 on feeding behaviour and timebudgets. In Period 2, grouped pigs slept more (P<0.01) and spent less time feeding (P<0.01) and rooting (P<0.01) than in Periods 1 and 3. The frequency of aggression decreased over time from mixing (P<0.001). Possible explanations for the changes in feeding behaviour when pigs are moved from individual to group housing are competition, group cohesion, or that the high frequency of feeder visits when the pigs are housed individually is a consequence of a lack of social stimulation. Of these different possibilities, the results suggest that group cohesion is most likely to have been causal in the observed changes in feeding behaviour.     

Basal metabolic rate is positively correlated with parental investment in laboratory mice

The assimilation capacity (AC) hypothesis for the evolution of endothermy predicts that the maternal basal metabolic rate (BMR) should be positively correlated with the capacity for parental investment. In this study, we provide a unique test of the AC model based on mice from a long-term selection experiment designed to produce divergent levels of BMR. By constructing experimental families with cross-fostered litters, we were able to control for the effect of the mother as well as the type of pup based on the selected lines. We found that mothers with genetically determined high levels of BMR were characterized by higher parental investment capacity, measured as the offspring growth rate. We also found higher food consumption and heavier visceral organs in the females with high BMR. These findings suggested that the high-BMR females have higher energy acquisition abilities. When the effect of the line type of a foster mother was controlled, the pup line type significantly affected the growth rate only in the first week of life, with young from the high-BMR line type growing more rapidly. Our results support the predictions of the AC model.     

Blood pressure and heart rate determination by radiotelemetry in mice

Mice are not commonly used in cardiovascular physiology, especially in space physiology because of methodological problems. The initial studies that have monitored arterial pressure and heart rate in mice used the tail cuff plethysmography method or classical catheterization techniques with a catheter liable to an external pressure transducer. But for long term arterial pressure measurements the studies have been facilitated by the development of radiotelemetry. This is a less constraining method as classical one allowing to monitor in continuous in freely moving animal blood pressure, heart rate and temperature. This technique allows to record these parameters thank to an implanted transmitter without physical connection with the monitoring system. The aim of this work was to valid the use of the radiotelemetry in mice to evaluate arterial blood pressure and heart rate during different stress conditions (but only control data are described in this paper).     

Evaluation of baroreflex control of heart rate in renovascular hypertensive mice

The objective of the present study was to evaluate the baroreflex and the autonomic control of heart rate (HR) in renovascular hypertensive mice. Experiments were carried out in conscious C57BL/6 (n = 16) mice 28 days after a 2-kidney 1-clip procedure (2K1C mice) or a sham operation (sham mice). Baroreflex sensitivity was evaluated by measuring changes in heart rate (HR) in response to increases or decreases in mean arterial pressure (MAP) induced by phenylephrine or sodium nitroprusside. Cardiac autonomic tone was determined by use of atropine and atenolol. Basal HR and MAP were significantly higher in 2K1C mice than in sham mice. The reflex tachycardia induced by decreases in MAP was greatly attenuated in 2K1C mice compared with sham mice. Consequently, the baroreflex sensitivity was greatly decreased (2.2 +/- 0.4 vs. 4.4 +/- 0.3 beats x min(-1) x mmHg(-1)) in hypertensive mice compared with sham mice. The reflex bradycardia induced by increases in MAP and the baroreflex sensitivity were similar in both groups. Evaluation of autonomic control of HR showed an increased sympathetic tone and a tendency to a decreased vagal tone in 2K1C mice compared with that in sham mice. 2K1C hypertension in mice is accompanied by resting tachycardia, increased predominance of the cardiac sympathetic tone over the cardiac vagal tone, and impairment of baroreflex sensitivity.     

The polyphyletic origin of laboratory inbred mice and their rate of evolution

The analysis of various genetic components in the standard inbred laboratory strains of mice shows that several taxonomic units of the complex species Mus musculus have contributed to their genetic background. These laboratory lines are often taken as archetypes of the mammalian genome and since their genealogy is known for around 80 years they have been used to estimate mutation rates and various other evolutionary parameters. The knowledge of their origins is important if one is to avoid erroneous interpretations. Do they possess haplotypes that could have existed in natural populations of M. musculus domesticus or are they the fruit of artificial recombinations between divergent genomes? We discuss this in the light of various genetic systems.     

The reduction of abnormal behaviors in individually housed rhesus monkeys (Macaca mulatta) with a foraging/grooming board

A new environmental enrichment device, termed a “foraging/grooming board,” was presented to 8 individually housed rhesus monkeys for the explicit purpose of reducing the level of aberrant behaviors manifested by these animals. The device, consisting of a piece of plexiglass covered with artificial fleece, had particles of food treats rubbed into it and was attached to the outside of each animal's home cage. All animals foraged from the board to the point that a significant reduction in the level of abnormal behavior was noted. Most animals also groomed the fleece covering the board, utilizing the same motor patterns that would be directed toward grooming another monkey. These boards are inexpensive to construct and easy to sanitize, and do not require placing animal facility personnel at risk to maintain them.