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.

Laboratory animal allergy is preventable in modern research facilities.

BACKGROUND: Historical data suggest 15% of laboratory animal workers develop IgE sensitisation and 10% symptoms of laboratory animal allergy (LAA), including occupational asthma. Individually ventilated cages (IVCs) are replacing conventional open cages; we sought to evaluate their impact on the development of LAA. METHODS: We surveyed 750 laboratory animal workers and measured airborne Mus m 1 (mouse allergen) levels in seven UK institutions. We compared the prevalence of sensitisation to mouse proteins (by specific IgE assay or skin prick test) and of work-related allergic symptoms in IVC-only and open cage units. RESULTS: Full-shift Mus m 1 levels were lower in IVC than open cage units (geometric mean 1.00 (95% CI 0.73-1.36) versus 8.35 (95% CI 6.97-9.95) ng·m-3; p<0.001), but varied eight-fold across the IVC units (geometric mean range 0.33-4.12 ng·m-3). Primary analyses on data from 216 participants with ≤3 years exposure to mice revealed a lower prevalence of sensitisation in those working in IVC units compared with conventional cage units (2.4% (n=2) versus 9.8% (n=13); p=0.052). Sensitisation in IVC units varied from 0% to 12.5%; the use of fitted respiratory protection was less common in IVC units where prevalence of sensitisation was higher. Work-related allergy symptoms were more frequently reported by mouse-sensitised individuals (46.7% versus 10.9%; p<0.001) and only by those working in open cage units. CONCLUSION: In contemporary practice, LAA is now largely preventable with the use of IVC systems and the judicious use of appropriate respiratory protection.

Urinary Peptides As a Novel Source of T Cell Allergen Epitopes.

Mouse allergy in both laboratory workers and in inner-city children is associated with allergic rhinitis and asthma, posing a serious public health concern. Urine is a major source of mouse allergens, as mice spray urine onto their surroundings, where the proteins dry up and become airborne on dust particles. Here, we tested whether oligopeptides that are abundant in mouse urine may contribute to mouse allergic T cell response. Over 1,300 distinct oligopeptides were detected by mass spectrometry analysis of the low molecular weight filtrate fraction of mouse urine (LoMo). Posttranslationally modified peptides were common, accounting for almost half of total peptides. A pool consisting of 225 unique oligopeptides of 13 residues or more in size identified within was tested for its capacity to elicit T cell reactivity in mouse allergic donors. Following 14-day in vitro stimulation of PBMCs, we detected responses in about 95% of donors tested, directed against 116 distinct peptides, predominantly associated with Th2 cytokines (IL-5). Peptides from non-urine related proteins such as epidermal growth factor, collagen, and Beta-globin accounted for the highest response (15.9, 9.1, and 8.1% of the total response, respectively). Peptides derived from major urinary proteins (MUPs), kidney androgen-regulated protein (KAP), and uromodulin were the main T cell targets from kidney or urine related sources. Further ex vivo analysis of enrichment of 4-1BB expressing cells demonstrated that LoMo pool-specific T cell reactivity can be detected directly ex vivo in mouse allergic but not in non-allergic donors. Further cytometric analysis of responding cells revealed a bone fide memory T cell phenotype and confirmed their Th2 polarization. Overall, these data suggest that mouse urine-derived oligopeptides are a novel target for mouse allergy-associated T cell responses, which may contribute to immunopathological mechanisms in mouse allergy.

Exposure of laboratory animal care workers to airborne mouse and rat allergens.

Urine of rats and mice is the main source of allergenic proteins that can enter the respiratory tract of laboratory animal care workers. Little is known about the levels and determinants of these exposures in the United States. We investigated the relationship between activities in animal facilities and levels of personal exposure to allergen by collecting personal breathing zone dust samples from 7 caretakers during full workdays for 1 wk. Mice and rat urinary allergens in inhalable dust were quantified via immunoassay. The activities of the sampled workers were observed, and the methods of preventing exposure to allergens were recorded. Mouse urinary allergen was detected in 20 of 39 measurements, yielding a geometric mean of 0.8 ng/m(3) with a maximum of 24 ng/m(3). Washing and cleaning cages and the number of mice handled daily were the most important determinants of personal exposure to mouse urinary allergen, as identified by using multiple linear regressions that explained 51% of total variance. Personal exposures to mouse urinary allergen were associated with day-to-day variation of tasks rather than characteristics of workers. Where potential for personal exposure is the highest, protective measures (N95 masks and cage dumping stations) appeared to be used, as is appropriate. Rat urinary allergen was detected in 4 of 39 measurements; detectable concentrations were between 0.8 and 39 ng/m(3). Only persons who handled rats were exposed to rat urinary allergen. The current findings are valuable for establishing exposure levels against which comparisons of improvement or deterioration of personal exposures can be made.

Comparison of urinary scents of two related mouse species, Mus spicilegus and Mus domesticus.

Whereas the house mouse (Mus domesticus) has been studied extensively in terms of physiology/behavior and pheromonal attributes, the evolutionarily related mound-building mouse (Mus spicilegus) has received attention only recently due to its divergent behavioral traits related to olfaction. To date, no chemical studies on urinary volatile compounds have been performed on M. spicilegus. The rationale for our investigations was to determine if there are differences in urinary volatiles of intact and castrated M. spicilegus males and to explore further whether this species could utilize the same or structurally similar pheromones as the male house mouse, M. domesticus. The use of capillary gas chromatography/mass spectrometry (GC-MS) together with sorptive stir bar extraction sampling enabled quantitative comparisons between the intact and castrated M. spicilegus urinary profiles. Additionally, through GC-MS and atomic emission (sulfur-selective) detection, we identified qualitative molecular differences between intact M. spicilegus and M. domesticus. A series of volatile and odoriferous lactones and the presence of coumarin were the unique features of M. spicilegus, as was the notable absence of 2-sec-butyl-4,5-dihydrothiazole (a prominent M. domesticus male pheromone) and other sulfur-containing compounds. Castration of M. spicilegus males eliminated several substances, including delta-hexalactone and gamma-octalactone, and substantially decreased additional compounds, suggesting their possible role in chemical communication. Some other M. domesticus pheromone components were also found in M. spicilegus urine. These comparative chemical analyses support the notion of metabolic similarities as well as the uniqueness of some volatiles for M. spicilegus, which may have a distinct physiological function in reproduction and behavior.

Dual sensitization to rat and mouse urinary allergens reflects cross-reactive molecules rather than atopy.

BACKGROUND: Sensitization to rats and mice can develop in laboratory animal workers exposed to only one species. Reasons for this dual sensitization are unclear but may reflect a genetic predisposition to developing allergy (atopy) or alternatively cross-reactivity between rat and mouse urinary allergens. We examined cross-reactivity between rat and mouse urine and the effect atopy has on dual sensitization in laboratory animal workers. METHODS: In a cross-sectional study the frequency of sensitization to rat and/or mouse was analysed in 498 employees exposed to both rat and mouse at work and 220 to rat only. RAST inhibitions, western blots and blot inhibitions were carried out on a subset of five individuals to assess cross-reactivity. RESULTS: Fourteen per cent of workers were sensitized to rats and 9% to mouse. Over half (62%) of rat sensitized individuals were also mouse sensitized and the majority (91%) of mouse sensitized individuals were also rat sensitized. IgE cross-reactivity was demonstrated between rat and mouse urine using RAST inhibitions. Rates of atopy did not differ between rat only sensitized individuals compared with those sensitized to both species. Sensitization to cats and rabbits was more common amongst those with dual sensitization. CONCLUSIONS: Dual sensitization to rat and mouse reflects IgE cross-reactivity rather than atopy. Individuals with dual sensitization are more likely to be sensitized to other animal allergens. These findings will have implications for individuals working with only one rodent species who develop sensitization and symptoms to be aware of the potential for allergy to other species.

Encoding choosiness: female attraction requires prior physical contact with individual male scents in mice.

Scents, detected through both the main and vomeronasal olfactory systems, play a crucial role in regulating reproductive behaviour in many mammals. In laboratory mice, female preference for airborne urinary scents from males (detected through the main olfactory system) is learnt through association with scents detected through the vomeronasal system during contact with the scent source. This may reflect a more complex assessment of individual males than that implied by laboratory mouse studies in which individual variation has largely been eliminated. To test this, we assessed female preference between male and female urine using wild house mice with natural individual genetic variation in urinary identity signals. We confirm that females exhibit a general preference for male over female urine when able to contact urine scents. However, they are only attracted to airborne urinary volatiles from individual males whose urine they have previously contacted. Even females with a natural exposure to many individuals of both sexes fail to develop generalized attraction to airborne male scents. This implies that information gained through contact with a specific male's scent is essential to stimulate attraction, providing a new perspective on the cues and olfactory pathways involved in sex recognition and mate assessment in rodents.

Social signals: the secret language of mice.

Animals are known to produce substances that modulate social and sexual behavior of conspecifics, but the mechanistic details underlying these phenomena have been elusive. A recent paper identifies a male-specific compound in mouse urine that activates olfactory bulb neurons and mediates behavioral attraction.

Detection of mouse and rat urinary aeroallergens with an improved ELISA.

BACKGROUND: Risk analysis of laboratory animal work presupposes allergen monitoring with sensitive methods. Commercial ELISA kits have recently become available for the detection of mouse (Mus m 1) and rat (Rat n 1) urinary allergen from settled dust samples and air samples with high allergen levels. OBJECTIVE: Our aims were to enhance the sensitivities of the commercial ELISA kits for low aeroallergen levels (less than 1 ng/m(3)) and to test these methods with air samples collected from an animal facility. METHODS: Personal and stationary air samples were collected from an animal facility during various tasks of laboratory animal work and from various premises of the animal facility. RESULTS: The sensitivities of the ELISA assays were improved with a careful choice of analysis parameters and reagents. The detection limits of 0.1 ng/m(3) for Mus m 1 and 0.8 ng/m(3) for Rat n 1 were established. The sensitized assays enabled detection of mouse and rat aeroallergens also from premises in which animals or dirty cages were not present during sampling. CONCLUSION: These sensitive assays will help to perform risk assessment in laboratory animal work. However, there remains a lack of standardized analytic procedures and occupational exposure limits for laboratory animal allergens.

[Postradiation volatile secretion of mice attractive for intact individuals]. / Attraktivnye dlia intaktnykh osobei postraditsionnye letuchie badeleniia myshei.

In early period after exposure to a sub-lethal dose (4 Gy) the exposed mice secreted volatile components (in their urine), which were attractive for intact individuals independently of genotype. The postradiation components were more attractive for allogene individuals than for singene ones. This reaction was different in exposed intact mice, which showed singene attractiveness of volatile secretion. Attractiveness of postradiation volatile components differed from the attractiveness of ones secreted after stress.

Hypersensitivity to molds in New York City in adults who have asthma.

Molds have been linked epidemiologically to asthma as a key aeroallergen in several studies. Other allergens such as cockroach have been linked to asthma in New York City (NYC). To our knowledge, however, the pattern of mold hypersensitivity has never been examined systematically in the NYC area. Thus, we sought to determine the association between mold hypersensitivity and asthma in a large group of ambulatory patients evaluated for allergic disease for the years 1993 through 2001 at a single medical center. Serological testing for mold-specific immunoglobulin E (IgE) as well as IgE specific for other aeroallergens was performed and the associations between allergen-specific IgE and the presence of asthma were examined using bivariate and multivariate analysis. Factor analysis showed that three distinct groupings of aeroallergen-specific IgE existed within the panel of allergens used. Group 1 consisted of cat dander and dust mites (Dermatophagoides farinae). Group 2 consisted of tree, grass, and ragweed pollen. Group 3 consisted of the Deuteromycetes molds, Alternaria tenuis, Aspergillus fumigatus, and Cladosporium herbarum. Patients with asthma had a highly significant increase in the incidence of hypersensitivity to cat/dust mites and to the molds. Multivariate analysis showed that the presence of hypersensitivity to either A. tenuis or C. herbarum had a significant independent association with asthma after adjustment for cat/dust mite hypersensitivity and after adjustment for other clinical factors. On the other hand, pollen hypersensitivity was not associated independently with asthma. Mold hypersensitivity was strongly correlated with hypersensitivity to cat or dust mites in patients who did not have asthma but not in patients who did have asthma. In the NYC area, recent pollen and spore counts show that mold spores are measurable in at least 75% of the year. Thus it is conceivable that mold hypersensitivity plays a contributing and independent role in initiating or perpetuating the allergic response in patients with asthma in the New York area.

Collection of mouse urine for bioassays.

Rodent bioassays often require fresh, uncontaminated urine from a large number of mice. The authors describe a quick and simple method to collect moderate to large amounts of urine from individual mice using disposable plastic trays.

Evaluation of individually ventilated cage systems for laboratory rodents: occupational health aspects.

New ventilated caging systems for laboratory animals were compared with conventional caging regarding allergen distribution, ergonomic suitability, cage environment and animal welfare. This paper presents occupational health evaluations. Mice were placed in individually ventilated cage (IVC) systems, a ventilated cabinet, and in cages on open shelves (conventional husbandry). The IVC systems were studied at negative and positive airflow. Aeroallergens were sampled on filters (n = 204, including controls) in undisturbed rooms and during cage changing. Concentrations of mouse urinary allergen (Mus m 1) in filter eluates were measured using sandwich ELISA. An ergonomic evaluation was performed with measurement of traction forces. Staff exposure during cage changing was high in all systems, range 116-4430 ng Mus m 1/m3. In undisturbed animal rooms, allergen levels were orders of magnitude higher when using conventional caging compared with ventilated systems; P < 0.001. At positive pressure both IVCs leaked allergen (median Mus m 1 concentration was < 0.08 ng/m3 at negative, but 6.5 ng/m3 (IVC1) and 0.8 ng/m3 (IVC2S) at positive pressure). The IVC systems had ergonomic disadvantages compared with the conventional husbandry and the ventilated cabinet, for instance with cages in unsuitable working heights. Ventilated husbandry solutions reduce levels of airborne allergen substantially at negative pressure, but are ergonomically less suitable. To prevent allergen exposure during cage changing, we propose that this procedure should be performed under ventilated conditions. Producers and users must cooperate in optimizing animal caging systems for both animals and staff.

Measurement of exposure to mouse urinary proteins in an epidemiological study.

OBJECTIVES: To develop an assay to measure airborne mouse urinary protein (MUP) and to assess the occupational exposure to MUP in the workforce of three establishments as part of an epidemiological study examining the influence of aeroallergen exposure on the development of allergic respiratory disease. METHODS: Personal air samples were collected from nine exposure groups during a workshift. A sensitive and reproducible competitive inhibition assay, which used rabbit antisera specific for MUP, was developed and used to measure the occupational exposure to MUP. RESULTS: The personal measurements of MUP showed that people with direct contact with mice (animal technicians) had the highest exposure followed in decreasing order by those working with anaesthetised animals or their tissue (postmortem workers and scientists) and those with indirect contact with mice (supervisors, office workers, and slide production workers). The only difference in concentrations of MUP between the three establishments were found for cage cleaners, which reflected differences in working practises for this exposure category. Air samples collected during the performance of specific tasks showed that high exposures to MUP were associated with handling mice, indirect contact with mice, and washing floors. CONCLUSIONS: Exposure to mouse urinary proteins has been measured in the occupational environment. This information can be used to determine the relation between exposure to MUP and the development of allergic and respiratory disease.

Epidermal growth factor in neonatal mouse urine: maturative effect of thyroxine.

Using a specific and sensitive epidermal growth factor (EGF) radioimmunoassay, we have shown measurable quantities of EGF in mouse urine during the neonatal period. Sephadex G-50 column chromatography demonstrated the presence of a single immunoreactive component at the position defined by standard EGF (mol wt 6045). Comparison of urine urea nitrogen and urine EGF levels in neonatal and adult mice showed adult values to be 3- and 16-fold higher, respectively. Kidney weights relative to body weight were similar in newborn and adult animals while kidney EGF concentration per mg protein was 2.5-fold higher in the adult. The relative submandibular gland (SMG) weight was slightly higher in adult female mice than in the newborn, whereas SMG-EGF concentration was 15,000-fold higher in the adult than in the newborn. Thyroxine administration to neonatal mice from day 0 to day 6 increased urine EGF concentration 7-fold compared to control pups. Though the hormone treatment elicited a significant increase in relative SMG weight, its EGF concentration like that of the kidneys remained unchanged. The results suggest that urine EGF is subject to thyroid hormone modulation in newborn animals and that the changes in urine EGF concentration are independent of changes in SMG and renal EGF levels.

Experience-based vocalization of male mice to female chemosignals.

Previous experimentation led to a suggestion that pituitary-mediated metabolites present in female urine are important determinants of stimulus potency in eliciting 70 kHz ultrasonic vocalizations from adult male mice (Mus musculus). In the present experiment this hypothesis was reexamined, taking into account the prior experience of the male test subjects. Male mice were given social experience with either hypophysectomized females or intact, normal females and were then monitored for ultrasonic vocalizations to chemosensory stimuli from both. Results indicated that males (1) will emit ultrasonic vocalizations to urine from hypophysectomized females and (2) tend to preferentially emit ultrasound to stimuli from the type of female previously encountered during the social experience regimen. It is proposed that the previously reported lack of ultrasound emission by males to chemosignals from hypophysectomized females can be accounted for via straightforward associative learning mechanisms.

False negative hyperglucosuria test-strip reactions in laboratory mice.

Inhibition of a variety of commercial test strips for hyperglucosuria was experienced in laboratory mice. All mouse strains tested were found to have sufficiently high levels of ascorbic acid to cause inhibition, and male levels were higher than those of females. A regime to obtain optimum detection of positive results is discussed.

Description of a disposable individual-mouse urine collection apparatus.

An inexpensive, individual mouse urine collection apparatus was constructed from a 250 ml conical polypropylene centrifuge tube and wire cloth. The construction did not require special tools, and was completed in 1 hour. This apparatus allowed the ad libitum feeding and watering of individual mice during the urine collection period. Volumes of urine collected ranged from 0.5-2.0 ml per 24 hours per 25 g mouse.

Water balance of small lactating rodents--III. Estimates of milk production and water recycling in lactating Mus musculus under various water regimes.

1. The exchanges of water between lactating female and young Mus musculus were modelled on the computer. 2. The model was used to estimate rates of milk production and water recycling in various litter sizes under various water regimes by following the time course of injected tritiated water. 3. The high correlation between estimated rates of milk production and actual growth rates of young was taken to indicate that the method gave if not the actual rates of milk production a very constant proportion of it. 4. Approximately 50% of the water secreted in milk is returned to the mother by recycling.