Microclimate in two types of rat cages

The microclimate in two types of rat cages (a Makrolon type IV with a solid floor and a stainless steel cage with a wire mesh floor (five rats per cage)) placed in the same macro-environment was compared. The temperature, relative humidity and ammonia concentration in the cages were measured twice a day for 8 days. The cages were cleaned every 4 days. The greatest difference between the cage types was in the ammonia build-up. In Makrolon cages the ammonia concentration never reached 5 ppm, whereas in steel cages it showed a constant increase and already exceeded the threshold limit for man (25 ppm for 8 h per day) on the third day after cleaning.     

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.     

Effects of cage density on behavior in young adult mice

Optimal housing conditions for mice can be achieved by minimizing environmental variables, such as those that may contribute to anxiety-like behavior. This study evaluated the effects of cage size on juvenile mice through assessment of differences in weaning weight, locomotor skills, and anxiety-like behavior. Eighteen pairs of male and pregnant female Swiss-Webster (Cr:SW) mice were housed in 3 different caging scenarios, providing 429, 505, or 729 cm2 of space. Litters were standardized to 10 pups per litter in each cage. Mice reared in each caging scenario were assessed with the open-field, light-dark exploration, and elevated plus-maze tests. No differences in weaning weight were noted. Mice reared in the 505- and 729-cm2 cages explored a significantly larger area of the open-field arena than did those in the 429-cm2 cages. Those reared in the 505-cm2 cages spent more time in the center of the open field than did those in the 729-cm2 cages, suggesting that anxiety-like behavior may be increased in the animals housed in the larger cages. This study did not establish a consistent link between decreased floor space and increased anxiety-like behavior; neither does there appear to be a consistent effect of available floor area on the development of locomotor skills on mouse pups.     

Trade-off between floor level and floor material in farmed silver foxes

Farmed silver foxes (Vulpes vulpes) were allowed to balance their known preference for an elevated floor against their presumed preference for a sand floor. In Experiment 1, foxes had to choose between two identical cages, connected with an opening. One cage had a wire floor and the other had a sand floor, but the cages either were on the same (low or elevated) or on different levels (one cage 40 cm higher than the other). In Experiment 2, the cage pairs were connected with a 1.2 m long wire-mesh tunnel, one cage was always on a higher level (50 cm) than the other. In Experiment 1, foxes always preferred the sand floor during their active time. They also preferred the sand floor for resting, if it was on the same level as wire floor, but did not show any genuine preference if the floors were on different levels. In Experiment 2, foxes never preferred the lower floor. They preferred the elevated sand floor for activity and the elevated wire floor for lying. When two floors were identical they preferred the elevated one. Their rest consisted of 11-22 bouts, a major part of these being spent in the preferred cage. They also preferred a previous lying site to a new one, often exclusively and independently of floor material. In Experiment 1 foxes preferred the sand floor whereas in Experiment 2 they preferred the elevated floor indicating that the ability of a trade-off situation to rank resources depends on the method it is inflicted.     

Effects of housing density and cage floor space on three strains of young adult inbred mice

Some recommendations in the Guide for the Care and Use of Laboratory Animals (the Guide) are based on best professional judgment. Our current efforts are directed toward replacement with data-driven standards. We demonstrated earlier that young adult C57BL/6J mice could be housed with half the floor space recommended in the Guide without discernable negative effects. This report extends that work by examining optimal housing densities for young adult male and female BALB/cJ, NOD/LtJ, and FVB/NJ mice. These 8-week studies were initiated with 3-week-old BALB/cJ and NOD/LtJ mice and 3- to 5-week-old FVB/NJ mice housed in three cage types. We adjusted the number of mice per cage to house them with the floor space recommended in the Guide (approximately 12 in2 [ca. 77 cm2] per mouse) down to 5.6 in2 [ca. 36 cm2] per mouse. Early-onset aggression occurred among FVB/NJ male mice housed at all densities in cages having 51.7 in2 (ca. 333 cm2) or 112.9 in2 (ca. 728 cm2) of space. FVB/NJ male mice housed in shoebox (67.6 in2 [ca. 436 cm2]) cages did not exhibit aggression until the fifth week. Urinary testosterone output was density-dependent only for BALB/cJ male mice in shoebox cages (output decreased with increasing density) and FVB/NJ male mice. We conclude that all but FVB/NJ male mice can be housed with half the floor space specified in the Guide. The aggression noted for male FVB/NJ mice may have been due to their age span, although this did not impact negatively on the female FVB/NJ mice.     

Olfactory discrimination of aliphatic odorants at 1 ppm: too easy for CD-1 mice to show odor structure–activity relationships?

Using an operant conditioning paradigm we tested the ability of CD-1 mice to discriminate between 25 odorants comprising members of five homologous series of aliphatic odorants (C4-C8) presented at a gas phase concentration of 1 ppm. We found (a) that all mice significantly discriminated between all 50 stimulus pairs that involved odorants sharing the same functional group, but differing in carbon chain length, as well as between all 50 stimulus pairs that involved odorants sharing the same carbon chain length but differing in functional group, (b) a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length with the acetic esters and the 2-ketones, but not with the 1-alcohols, n-aldehydes, and n-carboxylic acids tested, and (c) that odorant pairs differing in functional group were significantly better discriminated than odorant pairs differing in carbon chain length. These findings demonstrate that CD-1 mice have excellent discrimination ability for structurally related aliphatic odorants, that correlations between discrimination performance and structural similarity of odorants are odorant class-specific rather than a general phenomenon, and that both carbon chain length and type of functional group play an important role for odor quality coding in mice.     

A comparison of rodent caging systems based on microenvironmental parameters.

Four different mouse caging systems were evaluated for microenvironmental temperature, carbon dioxide, relative humidity (RH) and ammonia levels during a 7-day testing period. All caging systems evaluated had polycarbonate bases and consisted of either a molded polyester (MP) filter lid, one of two different polycarbonate filter lids, or no filter lid which served as a control. At 50% macroenvironmental RH (study I), all systems maintained an intracage temperature of 75.5 degrees F +/- 0.5 degrees. Both polycarbonate systems averaged greater than 2200 ppm of carbon dioxide more than the MP system and the controls. When compared with RH in the control cages, RH levels averaged over 20% and 5 to 8% RH greater in the polycarbonate filter lid systems and the MP system, respectively. There were no appreciable ammonia levels in either the MP or control systems. In the polycarbonate filter lid systems, ammonia levels were detectable on day 4 and were greater than 200 ppm by day 6. At 20% macroenvironmental RH (study II), there was a proportional 15 to 30% RH decrease from study I levels. Ammonia levels were undetectable in any system until day 7 and averaged only 17 ppm in one of the polycarbonate systems. Minimal differences were observed in studies III, IV and V when pine shavings were used instead of hardwood chips, a CD-1 stock instead of a DBA/2J strain, and different grades of filter inserts in the polycarbonate systems, respectively.     

The use of dirty bedding for detection of murine pathogens in sentinel mice

Sentinel Swiss (CD-1) mice, housed without filter bonnets, were seronegative for mouse hepatitis virus (MHV) for 8 consecutive months in an experimental colony of CD-1 mice. MHV titers had been detected sporadically in sentinel mice housed in this colony during a 2 year period. In an effort to determine whether MHV was still present in the colony, two methods of exposing sentinel mice to an animal room environment were compared under routine husbandry practices. Eight cages (12 mice per cage; 2 cages per rack) of experimental virus antibody free sentinel mice, housed without filter bonnets, were placed on the bottom shelf of 4 of 12 racks in the room. Twice each week, four cages of sentinel mice received a composite sample of dirty bedding (bedding used previously by mice in the room). The remaining four cages of experimental sentinels received fresh non-used bedding. Sentinel mice were bled at monthly intervals for MHV serology. After 4 months, mice from two cages which received dirty bedding seroconverted to MHV and mice from one cage were positive for Myobia musculi (mites). Three weeks later, all four cages of mice which received dirty bedding were positive for MHV and three were positive for mites. In contrast, only two of four cages of mice which received fresh bedding were positive for MHV and all were negative for mites. These findings indicate the importance of exposing sentinel mice to dirty bedding and that MHV and mites may go undetected for several months in a mouse colony when the incidence levels are low where standard sanitation procedures are used.     

Preferences of farmed blue foxes for different floor types

In the first experiment, farmed blue foxes were allowed to choose for 1 week between four standard farm cages equipped with different floor materials: plastic-coated wire mesh, dry wood, dry sand and wet (summer) or icy (winter) sand. Resting consisted of 10-15 separate bouts/day occupying 50-60% of the total 24-h. There were no other differences in the use of the cages except that the time spent on, and the duration and number of resting bouts were lower on wet or icy sand, resting periods being more affected than activity. In the second experiment, two cages were connected with a 1.2 m tunnel. One cage was always elevated (50 cm) compared to that one which was lower. One cage of each pair had a wire floor whereas the other cage either had sand or wire floor. Sand floor was preferred for active behaviours and wire floor for resting if these were on elevated level. Of two identical wire-floored cages, the elevated one had the priority. Foxes preferred to rest on the same floor where they had finished their previous resting bout, often exclusively and independently of floor material or floor level.     

Olfactory discrimination ability of CD-1 mice for aliphatic aldehydes as a function of stimulus concentration

Using an operant conditioning paradigm, we tested the ability of CD-1 mice to discriminate between members of a homologous series of aliphatic aldehydes presented at four different concentrations. We found that the mice were clearly capable of discriminating between all odorant pairs when stimuli were presented at concentrations of 1, 0.01, and 0.001 ppm (corresponding to four, two, and one log unit above the highest individual detection threshold) with no significant difference in performance between these concentrations. In contrast, the animals generally failed to discriminate above chance level when stimuli were presented at 0.0001 ppm (corresponding to the highest individual detection threshold) although stimuli were clearly detectable. Further, we found a significant negative correlation between discrimination performance and structural similarity of odorants in terms of differences in carbon chain length. These findings suggest that an increase in stimulus concentration of only one log unit above detection threshold appears to be sufficient for recruitment of additional subpopulations of odorant receptors to allow for qualitative recognition of aliphatic aldehydes.     

The effects of intracage ventilation on microenvironmental conditions in filter-top cages.

Filter-top cages, while effective in reducing cross contamination by particulate material including microbes, can also cause accumulation of the waste gases carbon dioxide and ammonia as well as increased intracage relative humidity. A prototype system which provided each cage with 23 air changes per hour through a nozzle inserted in the filter lid was evaluated. The ventilated cageing system was effective in reducing intracage carbon dioxide, ammonia and relative humidity levels. Mean weekly carbon dioxide levels were 2900 ppm lower, ammonia levels 240 ppm lower and intracage relative humidity levels 8% lower in the ventilated cages than in unventilated controls.     

Modulatory Effects of Testosterone on 1-Methyl-4-Phenyl- 1, 2, 3, 6-Tetrahydropyridine-Induced Neurotoxicity

Abstract: In this experiment, we examined the modulatory effects of testosterone on the parkinsonism-inducing drug 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) in two strains of mice. Orchidectomized male CD-1 and C57/BI mice were implanted with either empty Silastic capsules or capsules containing testosterone and subsequently treated with MPTP. A small area of the corpus striatum was removed for determination of dopamine (DA) content, whereas the remainder was superfused and used to measure L-DOPA (5 μ M )-evoked DA release. In animals treated with MPTP, L-DOPA-evoked DA release was reduced significantly in CD-1 mice, but not in C57/BI mice, treated with testosterone. No differences in L-DOPA-stimulated DA release between MPTP- versus vehicle-treated mice was observed in either the CD-1 or C57/BI mice receiving empty Silastic capsules. Corpus striatum DA contents were more severely depleted in the MPTP-sensitive C57/BI versus the CD-1 mouse strain irrespective of hormone treatment. These results confirm previous results demonstrating differences in these two mouse strains in response to the neurotoxic effects of MPTP upon corpus striatum DA content. More interestingly, they show an important differential modulatory effect of testosterone upon L-DOPA-evoked DA release as a function of MPTP treatment and indicate that testosterone significantly alters the neurotoxic effects of MPTP in the CD-1 mouse.     

Effects of housing density and cage floor space on C57BL/6J mice

The Guide for the Care and Use of Laboratory Animals (the Guide) is widely accepted as the housing standard by most Institutional Animal Care and Use Committees. The recommendations are based on best professional judgment rather than experimental data. Current efforts are directed toward replacing these guidelines with data-driven, species-appropriate standards. Our studies were undertaken to determine the optimum housing density for C57BL/6J mice, the most commonly used inbred mouse strain. Four-week-old mice were housed for 8 weeks at four densities (the recommended approximately 12 in2 [ca. 77.4 cm2]/mouse down to 5.6 in2 [ca. 36.1 cm2]/mouse) in three cage types with various amounts of floor space. Housing density did not affect a variety of physiologic parameters but did affect certain micro-environmental parameters, although these remained within accepted ranges. A second study was undertaken housing C57BL/6J mice with as little as 3.2 in2/mouse (ca. 20.6 cm2). The major effect was elevated ammonia concentrations that exceeded limits acceptable in the workplace at increased housing densities; however, the nasal passages and eyeballs of the mice remained microscopically normal. On the basis of these results, we conclude that C57BL/6J mice as large as 29 g may be housed with 5.6 in2 of floor space per mouse. This area is approximately half the floor space recommended in the Guide. The role of the Guide is to ensure that laboratory animals are well treated and housed in a species-appropriate manner. Our data suggest that current policies could be altered in order to provide the optimal habitation conditions matched to this species' social needs.     

The impact of reduced frequency of cage changes on the health of mice housed in ventilated cages

Our purpose in this investigation was to determine if we could reduce cage changing frequency without adversely affecting the health of mice. We housed mice at three different cage changing frequencies: 7, 14, and 21 days, each at three different cage ventilation rates: 30, 60 and 100 air changes per hour (ACH), for a total of nine experimental conditions. For each condition, we evaluated the health of 12 breeding pairs and 12 breeding trios of C57BL/6J mice for 7 months. Health was assessed by breeding performance, weanling weight and growth, plasma corticosterone levels, immune function, and histological examination of selected organs. Over a period of 4 months, we monitored the cage microenvironment for ammonia and carbon dioxide concentrations, relative humidity, and temperature one day prior to changing the cage. The relative humidity, carbon dioxide concentrations, and temperature of the cages at all conditions were within acceptable levels. Ammonia concentrations remained below 25 ppm (parts per million) in most cages, but, even at higher concentrations, did not adversely affect the health of mice. Frequency of cage changing had only one significant effect; pup mortality with pair matings was greater at the cage changing frequency of 7 days compared with 14 or 21 days. In addition, pup mortality with pair matings was higher at 30 ACH compared with other ventilation rates. In conclusion, under the conditions of this study, cage changes once every 14 days and ventilation rates of 60 ACH provide optimum conditions for animal health and practical husbandry.     

Age-associated changes in plasma testosterone levels in male mice and their relation to social dominance or subordinance

Plasma testosterone (T) levels were determined in male mice of the CD-1 (ICR) strain from 30 to 680 days of age. The mean plasma T was 5.2 ± 1.0 ng/ml for adult male mice (70–400 days old, 102 mice) and 1.8 ± 0.9 ng/ml for old males (450–680 days old, 11 mice). Through 140 determinations, the highest T value obtained was 52.3 ng/ml in an 80-day-old individual while the lowest was 0.08 ng/ml in a 680-day-old animal. It was found that plasma T was significantly decreased in the old age, although individual variation in T levels was considerable. On the other hand, social dominance or subordinance was examined among male mice in each cage and its relation to plasma T was investigated. Dominance or subordinance was verified by frequency of chases or attacks delivered or received, or number of fights won or lost. It was observed that dominant-subordinate relationships were present in 6 out of 10 cages and that the dominant individual had a significantly higher T level than the subordinate male; the mean T level was 10.5 ± 2.5 ng/ml for the dominant individuals vs 2.2 ± 0.8 ng/ml for the subordinate ones. Marked individual variation in plasma T titers in male mice seems to be related to the dominance/subordinance rank within a group.     

Modulatory Effects of Mild Carbon Monoxide Exposure in the Developing Mouse Cochlea

Carbon monoxide (CO) is well known as a highly toxic poison at high concentrations, yet in physiologic amounts it is an endogenous biological messenger in organs such as the internal ear and brain. In this study we tested the hypothesis that chronic very mild CO exposure at concentrations 25-ppm increases the expression of oxidative stress protecting enzymes within the cellular milieu of the developing inner ear (cochlea) of the normal CD-1 mouse. In addition we tested also the hypothesis that CO can decrease the pre-existing condition of oxidative stress in the mouse model for the human medical condition systemic lupus erythematosus by increasing two protective enzymes heme-oxygenase-1 (HO-1), and superoxide dismutase-2 (SOD-2). CD-1 and MRL/lpr mice were exposed to mild CO concentrations (25 ppm in air) from prenatal only and prenatal followed by early postnatal day 5 to postnatal day 20. The expression of cell markers specific for oxidative stress, and related neural/endothelial markers were investigated at the level of the gene products by immunohistochemistry, proteomics and mRNA expression (quantitative real time-PCR). We found that in the CD-1 and MRL/lpr mouse cochlea SOD-2 and HO-1 were upregulated. In this mouse model of autoimmune disease defense mechanism are attenuated, thus mild CO exposure is beneficial. Several genes (mRNA) and proteins detected by proteomics involved in cellular protection were upregulated in the CO exposed CD-1 mouse and the MRL/lpr mouse.     

Olfactory sensitivity for enantiomers and their racemic mixtures--a comparative study in CD-1 mice and spider monkeys

Using a conditioning paradigm, the olfactory sensitivity of six CD-1 mice for the enantiomers of carvone and of limonene as well as for their racemic mixtures was investigated. With all six stimuli, the animals significantly discriminated concentrations 相似文献   

Design and construction of cage environments for air ion and electric field research

This report describes the design and construction of cage environments suitable for chronic exposures of large groups of mice to air ions and electric fields. These environments provide defined and reproducible ion densities, ion flux, DC electric fields, sound levels, air temperature and air quality. When used during a 2 year study, these cage environments served as a durable and reliable continuous exposure system. Three environmental chambers (cubicles) housed a total of 12 cages and provided control of air temperature, air purity and lighting. Exposure cages had grounded metal exterior walls, a plexiglass door and interior walls lined with formica. An internal isolated field plate supplemented with guard wires, energized with ca 1000 VDC, created about a 2 kV/m electric field at the grounded cage floor. Air ions resulted from the beta emission of sealed tritium foils mounted on the field plate. Cages provided high ion (1.3×10⁵ ions/cc), low ion (1.6×10³ ions/cc) and field only (ion depleted < 50 ions/cc) conditions for both polarities with similar electric fields in ionized and field only cages. Detailed mapping of the floor level ion flux using 100 cm² flat probes gave average fluxes of 880 fA cm^–2 in high ion cages and 10 fA cm^–2 in low ion cages. Whole body currents measured using live anesthethized mice in high ion cages averaged 104±63 pA. Both ion flux and whole body currents remained constant over time, indicating no charge accumulation on body fur or cage wall surfaces in this exposure system.     

Forest leaf litter decomposition in the vicinity of a zinc smelter

Summary Concentrations of about 26,000 ppm Zn, 10,000 ppm Fe, 2,300 ppm Pb, 900 ppm Cd, 340 ppm Cu, and 0.40% S were measured in the O₂ litter horizon about 1 km from a zinc smelter at Palmerton, Pennsylvania. Samples taken about 6 km east of the smelter had concentrations of about 15,000 ppm Zn, 6,500 ppm Fe, 970 ppm Pb, 250 ppm Cd, 170 ppm Cu, and 0.26% S. Samples from a control area about 40 km east of the smelter had concentrations of 2,800 ppm Fe, 650 ppm Zn, 260 ppm Pb, 50 ppm Cu, 9 ppm Cd, and 0.13% S.Litter bags were used to estimate first-year weight loss in sassafras leaves and a mixture of chestnut oak/red oak leaves in the three sites. At the end of one year, average weight loss for sassafras was 39.3% in the control site, 21.8% at 6 km, and 17.5% at the 1 km site. For the chestnut oak/red oak mixture, average weight loss was 36.8% (40 km), 25.7% (6 km), and 19.1% (1 km). Numbers and diversity of soil microarthropods inhabiting the litter bags showed a corresponding decline at sites near the smelter. Concentrations of Ca, Cd, Cr, Cu, Fe, Mg, Mn, N, Na, Ni, P, Pb, S and Zn in the decomposing litter were also measured.The average amount of organic matter on the forest floor was estimated to be 3.8 kg/m² in the control site, about 3.8 kg/m² at 6 km, and about 8.1 kg/m² 1 km from the smelter. Average thickness of the litter horizons in these three sites was 6.0 cm (40 km), 7.0 cm (6 km), and 12.4 cm (1 km), suggesting a long-term depression of decomposition and mineral cycling near the smelter.     

DNA adducts of styrene-7,8-oxide in target and non-target organs for tumor induction in rat and mouse after repeated inhalation exposure to styrene

Styrene by inhalation had been shown to increase the lung tumor incidence in mice at 20 ppm and higher, but was not carcinogenic in rats at up to 1000 ppm. Styrene-7,8-oxide, the major metabolic intermediate, has weak electrophilic reactivity. Therefore, DNA adduct formation was expected at a low level and a 32P-postlabeling method for a determination of the two regioisomeric 2'-deoxyguanosyl-O6-adducts at the alpha(7)- and beta(8)-positions had been established. The first question was whether DNA adducts could be measured in the rat at the end of the 2 years exposure of a bioassay for carcinogenicity, even though tumor incidence was not increased. Liver samples of male and female CD rats were available for DNA adduct analysis. Adducts were above the limit of detection only in the highest dose group (1000 ppm), with median levels of 9 and 8 adducts per 10(7) nucleotides in males and females, respectively (sum of alpha- and beta-adducts). The result indicates that the rat liver tolerated a relatively high steady-state level of styrene-induced DNA adducts without detectable increase in tumor formation. The second question was whether different DNA adduct levels in the lung of rats and mice could account for the species difference in tumor incidence. Groups of female CD-1 mice were exposed for 2 weeks to 0, 40, and 160 ppm styrene (6h per day; 5 days per week), female CD rats were exposed to 0 and 500 ppm. In none of the lung DNA samples were adducts above a limit of detection of 1 adduct per 10(7) DNA nucleotides. The data indicate that species- and organ-specific tumor induction by styrene is not reflected by DNA adduct levels determined in tissue homogenate. The particular susceptibility of the mouse lung might have to be based on other reactive metabolites and DNA adducts, indirect DNA damage and/or cell-type specific toxicity and tumor promotion.