Airborne differences in odors emitted byRattus norvegicus in response to reward and nonreward

To test the hypothesis that rats (Rattus norvegicus) emit airborne, differential odors in response to reward and nonreward, donor rats received random sequences of rewarded and nonrewarded placements in small compartments and an airstream transported odors from these compartments to test rats in a separate chamber. When donors remained in the compartments during, or were removed just prior to, air transport, test rats utilized transported odors as discriminative cues signaling their own reward and nonreward for a lever-press response. When the airstream was passed through a clean compartment containing paper flooring extracted from donor compartments, test rats were not able to discriminate. Test trials to assess for control by food odors suggest that donor-produced odors, rather than food odors per se, provided the discriminative signals for test rats. Results confirm the existence of somewhat volatile, although apparently stable, odors emitted in response to reward and nonreward, and implicate a differential in amount and/or type of odor produced by donors to these two events as the source of discriminative control.     

Use of odor baits in traps to test reactions to intra- and interspecific chemical cues in house mice living in outdoor enclosures

House mice (Mus musculus) living in outdoor enclosures were tested for urinary chemical cue preferences using odor-baited traps. In the first experiment, with only volatile cues available, odors from conspecific males and females of various age classes and reproductive conditions were tested; no preferences were exhibited. In the second experiment mice had both nonvolatile and volatile cues available from the same sources as in experiment I. All age and sex class and female reproductive condition groups exhibited odor cue preferences except juvenile females. There were no specific odor cue preferences exhibited by any of the responder types with regard to odors from juvenile females. In the third and fifth experiments, mice were presented with nonvolatile plus volatile or only volatile urine odor cues, respectively, from four genera,Mus, Peromyscus, Microtus, andHomo. Mice of all age classes and both sexes preferredMus musculus odor, were neutral towardMicrotus ochrogaster odor, and avoided odors fromPeromyscus leucopus andHomo sapiens; these patterns were the same regardless of whether only volatile or both volatile and nonvolatile cues were presented. The fourth and sixth experiments involved testing volatile cues only and volatile cues plus nonvolatile cues from human sweat or feces from dogs, cats, or shrews. Mice avoided the human sweat and feces from cats and shrews, but were neutral toward the odor of dog feces. There were effects on whether mice were trapped in the interior of the enclosure or on the perimeter for some odors tested in these six experiments. The findings provide insights regarding possible functions of odor cues in the behavioral ecology of house mice. Odor-baiting traps can be an effective tool with respect to testing some, but not all questions pertaining to olfactory cues and house mouse social biology.     

Urine-marking and the response to fresh vs. aged urine in wild and domestic Norway rats

Wild and domestic Norway rats (Rattus norvegiens) were compared in regard to their tendency to investigate the odors of fresh vs. aged rat urine and to urine-mark metal rods and wooden blocks placed in their home cages. Castration, sex, and domestication had no effect on the tendency to investigate sources of fresh vs. aged urine odors, but the odor of aged urine was more attractive than fresh urine for most subjects tested. The frequency of urine marking was lower for females and castrated males but generally did not differ between wild and domestic stocks. The implications of these findings for the ecology of the species are discussed.     

Responses of Wild Norway Rats (Rattus norvegicus) to Predator Odors

If rats could be shown to avoid the odors of predators, then conservation managers could manipulate this behavior to exclude rats from important conservation sites. We evaluated the ability of six predator odors to elicit avoidance responses by wild-caught Norway rats (Rattus norvegicus) from two New Zealand populations (Kapiti Island and North Island). Kapiti Island is free of mammalian predators, while the North Island has established populations of felids, canids, and mustelids. Three of the predator odors were real and three were synthesized volatile ingredients of real animal feces or urine. We compared the rats' responses to predator odors with their responses to three natural herbivore odors. We used a Y maze, and rats were offered the choice of an odor in one arm of the maze and distilled water in the other arm. Each choice arm was ducted at the entrance to remove air and the odor. We recorded: (1) the time until the rat left the first arm of the maze, (2) the time until the rat visited each arm, (3) the number of visits to each arm, (4) the amount of time spent in each arm, and (5) a total activity score for each rat. Kapiti rats showed an aversion to five of the six predator odors, despite never having encountered them before. Kapiti rats visited herbivore odors more often than carnivore odors and were less active in the presence of carnivore odors than they were when tested with herbivore odors. In addition, Kapiti rats approached some herbivore odors more quickly than they approached carnivore odors. North Island rats appeared to avoid cat urine, but despite being predator experienced, did not show a consistent aversion to the carnivore odors we tested. Both samples of rats, but particularly the North Island group, showed high individual variation. We conclude that predator odors are unlikely to be an effective management tool for all populations of this species of rat because of this high individual variability and the likelihood that each island population will differ in its responses to a given odor.     

Mice Respond Differently to Urine and Its Major Volatile Constituents from Male and Female Ferrets

Our previous chemical investigation showed that the concentrations of urinary volatiles from males were much higher than those from females in the ferret (Mustela furo). The current study was designed to examine the behavioral significance and ecological relevance of this difference for one of the main prey of the ferret, the house mouse (Mus musculus). Our data showed that male mice displayed no difference in their response to raw male and female ferret urine. However, they showed significantly less response to female mouse urine mixed with ferret urine than to pure female mouse urine, and to female mouse urine mixed with male ferret urine than to female mouse urine mixed with female ferret urine. Furthermore, high levels of the three major volatiles (quinoline, 2,5-dimethylpyrazine, and 4-hepatanone) in male ferret urine were as effective as raw male ferret urine was in inhibiting the response of male mice. We discuss the ecological and behavioral significance of these findings in terms of chemical mimicry and cognitive feature extraction of predator odors in mice.     

Laboratory and field evaluation of predator odors as repellents for kiore (Rattus exulans) and ship rats (R. rattus)

Predator odors may serve to stop rats from entering conservation areas or to decrease predation, food consumption, and other damage by rats in areas tainted with predator odor. We compared the efficacy of real predator odors and synthetic odors (derived from the urine and feces of carnivores) as rat repellents with real herbivore odors as controls in a Y maze. We tested six predator odors: cat (Felis catus) urine and feces, mongoose (Herpestes auropunctatus) feces, n-propylthietane, S-methyl, methyl butanol, and isopentyl-methyl sulphide. The herbivore odors we used were: red deer (Cervus elaphus) urine, guinea pig (Cavia porcellus) feces, and white rabbit (Oryctolagus cuniculus) urine. Ship rats (Rattus rattus) and kiore or Polynesian rats (R. exulans) showed no aversion to any of the six predator odors when compared with herbivore odors. Ship rats, however, may have avoided synthesized odors more than real ones. We applied two odors (S-methyl, methyl butanol and n-propylthietane) to purpose-built feeders in native forest but recorded no change in either visitation rate or duration of visits for rodents [rats and mice (Mus musculus)] or possums (Trichosurus vulpecula). The consumption of maize at feeders was correlated with the number and duration of possum visits, but only weakly correlated with the number of visits by rodents. Consumption of maize was unaffected by the odor associated with the feeder. It is unlikely that the odors we tested will be useful in deterring rodents or possums from areas where they have been removed for economic, public health or conservation reasons.     

An investigation of human apocrine gland secretion for axillary odor precursors

Recently completed studies from our laboratories have demonstrated that the characteristic human male axillary odors consist of C₆ to C₁₁ normal, branched, and unsaturated aliphatic acids, with (E)-3-methyl-2-hexenoic acid being the most abundant. To investigate the mechanism by which the odor is formed, it is necessary to determine the nature of the odorless precursor(s) found in the apocrine secretion which is converted by the cutaneous microorganisms to the characteristic axillary odor. Pooled apocrine secretion was obtained from several male volunteers by intracutaneous injection of epinephrine. Partitioning this secretion into aqueous and organic soluble fractions was followed by hydrolysis of each fraction with NaOH or incubation with axillary microorganisms (cutaneous lipophilic corynebacterium). Analysis by gas chromatography/mass spectrometry (GC/MS) revealed the presence of (E)- and (Z)-3-methyl-2-hexenoic acid in the aqueous phase hydrolysate and aqueous phase incubated with bacteria; however, only a trace amount was seen in the resultant organic phase mixtures. These results suggest that a water-soluble precursor(s) is converted by the axillary flora to the characteristic axillary odors.     

Use of Chemical Communication by the Subterranean Rodent Ctenomys talarum (Tuco-Tuco) During the Breeding Season

Solitary subterranean rodents with a low frequency of direct contact between conspecifics are expected to use chemical communication to coordinate social and reproductive behavior. We examined whether reproductive tuco-tucos (Ctenomys talarum) were able to discriminate the reproductive condition, sex, and source population of conspecifics by means of chemical cues contained in urine, feces, soiled shavings, or anogenital secretions. During preference tests in which animals had direct contact with these chemical cues, tuco-tucos were able to determine the reproductive condition of opposite sex conspecifics independent of the source of odor. When only olfactory cues were available, both sexes discriminated reproductive condition of opposite sex individuals using urine. Females were also able to discriminate the reproductive condition of males using soiled shavings. Females spent more time investigating male odors than female odors; except in the case of feces, breeding males spent similar amounts of time investigating male and female odors. No preferences were detected for opposite sex urine from members of an animal's own versus another population. The role of chemical cues in territory defense and breeding performance by this highly territorial subterranean rodent is discussed.     

Responses to inter- and intraspecific scent marks in pine martens (Martes martes)

In order to analyze intra and interspecific olfactory discrimination, behavioral responses (sniffing and marking) towards various odors were observed in pine martens (Manes martes). Two adult males and two adult females were tested for intraspecific discrimination of abdominal gland odors and urine. Both sexes sniffed and marked objects carrying their own scent less than unscented objects. There were no differences in sniffing or marking objects impregnated with odors from known and unknown conspecifics of the opposite sex or objects carrying the odor of an unknown male or female. A second experiment with two adult females gave no evidence for interspecific discrimination: no differences emerged when comparing reactions towards marks of pine martens, stone martens (a closely related species), or genets. The most obvious result of this study is the reduced response of pine martens towards their own mark. It is suggested that scent marking in martens may reflect autocommunication, the primary effect being to familiarize an animal with its environment.     

Odor significance of undesirable degradation compounds in heated triolein and trilinolein

To better understand production of undesirable or negative odors such as fruity, plastic, and waxy that are characteristic of higher oleic acid-containing oils, model heated oil systems of triolein and trilinolein were studied. Identification of the odor significance of volatile compounds produced by fractionated and nonfractionated triolein and trilinolein was done by purge and trap-gas chromatography-ion trap mass spectrometry-olfactometry. The predominant odors of the triolein heated 1, 3, and 6 h at 190°C were fruity and plastic, with other negative odors of acrid and grassy. Some of the volatile compounds that produced negative odors in heated triolein, in order of increasing concentration, were hexanal (grassy), octanal (fruity), (E)-2-decenal (plastic), nonanal (fruity), and (E)-2-undecenal (plastic). Some of the negative odor compounds in trilinolein heated for 1, 3 and 6 h, in order of increasing concentration, included (E)-2-nonenal (plastic), pentanal (grassy), and hexanal (grassy). However, the amount of volatile compounds produced and the intensity levels of the odors were lower in trilinolein than in triolein. Formation of many of the volatiles was explained after identification of the volatile precursors, including epoxy, keto, and dimer oxidation products that were produced during heating. Presented at the American Oil Chemists’ Society 90th Annual Meeting & Expo, Orlando, FL, May 9–12, 1999.     

Effect of predator odors on heart rate and metabolic rate of wapiti (Cervus elaphus canadensis)

We measured the heart rate (HR) and oxygen consumption ( ) of wapiti (Cervus elaphus canadensis) before, during, and after presentation of biologically irrelevant odors (pentane, thiophene, and a perfume), artificial predator odors (an ether extract of cougar feces, and PDT, a compound found in mustelid anal gland secretion), stale predator odors (dog feces and urine and fox urine, kept at ambient temperature for a few weeks), and fresh predator odors (wolf, coyote, and cougar feces and the odor of a dead coyote, kept frozen between collection and test). Overall, responses to odors were small compared to other stressful stimuli. Individual variability was high among scents and among wapiti, but two of the fresh predator odors (cougar and wolf feces) produced larger HR and responses than the other scents and were more often successful at producing responses. As a group, fresh predator odors produced larger tachycardias and elicited a larger number of significant HR responses than biologically irrelevant novel odors. although the two classes of odors did not differ in their effect on . Although several other studies have shown that ungulates have reduced feeding levels when their food is scented with predator odors, it is not clear if this is due to reduced palatability or antipredator behavior. This study is the first demonstration that a wild ungulate species reacted more strongly to predator odors than to other odors in a nonfeeding situation.     

Odor similarity between stress-inducing odorants in wistar rats

Odor similarity to the odor of 4-mercapto-4-methyl-2-pentanone (I) was measured on eight rats by generalization of a conditioned avoidance response tocis- andtrans-8-mercapto-p-menthan-3-one (II and III), 3-mercapto-3-methyl-2-pentanone (V), andt-amyl mercaptan (VI). Previously, these odorants had been found to induce stress in rats in an open-field situation. In the present experiment, rats generalized the avoidance response learned with I, for V, VI, and to a lesser extenttrans-isomer III, implying odor similarities;cis isomer II was discriminated. Odor similarity between mercapto ketone I and mercaptan VI is surprising since VI lacks the keto group.     

Demonstration of an odorous intramale primer effect in short-tailed vole,Microtus agrestis L.

Anal (proctodeal) glands of maleMicrotus agrestis housed in social isolation undergo severe atrophy. Their weight and volume is significantly lower than those of the stock control males. The atrophied glands can be revived by subjecting deprived voles to various social odors. Atrophied glands of isolated males do not respond to the odors of male and female urine, voided feces of females, and unvoided feces of males. Atrophied anal glands of males exposed to voided male feces (which have passed the orifice of the anal gland) and soiled bedding from adult males show strong recrudescence. The mean weight and volume of the glands and plasma testosterone level are significantly higher than of males maintained in complete social isolation, although they are significantly less than those of stock control males. Atrophied glands of socially deprived males strongly respond to the odor of ethereal extract of gland secretion. In males exposed daily to anal gland secretion extract, the weight and volume of the gland and plasma testosterone level increase and are not significantly different from those of stock controls. They enjoy higher plasma testosterone levels and consequently larger and more active anal glands than complete isolates.     

Species-specific Expression of Major Urinary Proteins in the House Mice (<Emphasis Type="Italic">Mus musculus musculus</Emphasis> and <Emphasis Type="Italic">Mus musculus domesticus</Emphasis>)

The analysis of expression of pheromone-carrying major urinary proteins (MUPs) from two subspecies of house mice (Mus m. musculus, Mus m. domesticus) was studied. It has been previously shown that commensal populations of the two subspecies can discriminate on the basis of urinary signals. MUPs are predominant urinary proteins that protect pheromones from rapid degradation in a hydrophilic environment, and individuals of M. m. musculus tend to rely on these urinary cues in the process of subspecies discrimination more than M. m. domesticus individuals. Although it is not precisely known what triggers phenotypic and epigenetic changes of MUP expression, our results show that in the subspecies M. m. musculus, sex is a significant factor influencing variations in the regulation of selected MUPs in the liver. Furthermore, male M. m. musculus individuals expressed all the studied MUPs’ mRNA significantly more than females or individuals of either sex in M. m. domesticus. Correspondingly, the pattern of mRNA abundance was corroborated with the level of total MUP concentration in the urine, such that the level of sexual dimorphism was also significant and species-specific. Our finding introduces a hypothesis that quantitative variation of these proteins may be an essential part of a subspecies recognition system that maintains homospecific mixing.     

Sex-identifying urine and molt signals in lobster (Homarus americanus)

During courtship, premolt female lobsters,Homarus americanus, choose a male and initiate a pair bond by repeated approaches to his shelter. The male allows such a female to share his shelter for about one week. This knowledge formed the basis to search for quantitative evidence for lobster sex pheromone(s) used in courtship: male cues to allow premolt females to identify a preferred male, and female cues to allow males to identify a premolt mature female. In each of four 1500-liter naturalistic aquaria, the behavioral responses of one female and two male lobsters to male and female lobster urine (0.5 ml) and body odor (20 ml) stimuli were observed. These stimuli were injected once or twice per day into a continuously flowing delivery tube attached to lobster shelters. Habituation to stimulus introduction—a serious problem in earlier experiments—was apparently avoided in the more natural social and physical environment we employed in these experiments. We demonstrated that male and female molt body odors contain different chemical substances: females responded to male molt body odor and males responded to female molt body odor but not vice versa. In general, male and female intermolt urine caused strong responses; however, females responded only weakly to male urine. This suggests that male and female urine are chemically different. Female urine and molt body odor caused a typically male high-on-legs response. These results show that molt body odors and intermolt urine contain sex-specific substances, which may be used in lobster courtship as well as other social interactions.     

Laboratory Evaluation of Predator Odors for Eliciting an Avoidance Response in Roof Rats (Rattus rattus)

We evaluated eight synthetic predator odors and mongoose (Herpestes auropunctatus) feces for eliciting avoidance responses and/or reduced feeding by wild captured Hawaiian roof rats (Rattus rattus). In a bioassay arena, we recorded: (1) time until each rat entered the arena, (2) time elapsed until first eating bout, (3) time spent in each half of the arena, (4) number of eating bouts, and (5) consumption. Rats displayed a response to the predator odors in terms of increased elapsed time before initial arena entry and initial eating bout, a lower number of eating bouts, and less food consumption than in the respective control groups. The odor that produced the greatest differences in response relative to the control group was 3,3-dimethyl-1,2-dithiolane [from red fox (Vulpes vulpes) feces and mustelid anal scent gland]. Mongoose fecal odor produced different responses in four of the five variables measured while (E,Z)-2,4,5-trimethyl-³-thiazoIine (red fox feces) and 4-mercapto-4-methylpentan-2-one (red fox urine and feces) odors were different from the control group in three of the five variables measured. These laboratory responses suggest that wild Hawaiian roof rats avoid predator odors.     

Odors in traps: Does most recent occupant influence capture rates for house mice?

Responses of house mice (Mus domesticus) to odors in live traps were studied in a series of eight 0.1-ha outdoor field enclosures. It was assumed that the most recent mouse capture would provide the predominant odor in a trap for at least one week. Three different populations were tested, one in 1989 and two in 1992, involving over 800 different mice. Similar response patterns were recorded from all three groups. Two types of questions were tested: (1) Were there any biases contingent upon what had been previously caught? (2) Were there consistent responses of mice of particular age, sex, or reproductive classes to trap odors? Traps soiled by juvenile females caught adult females significantly less often than expected, but there were no consistent relationships in terms of the effects of specific residual odors on the subsequent capture at a particular trap. For various age, sex, and reproductive classes, (1) adult males preferred odors from juvenile and estrous females and avoided odors of other males significantly more than expected, (2) juvenile females selected traps with odors of other juvenile females and avoided all other types of female odors significantly more than expected, (3) nonestrous females exhibited a significant preference for adult male odor, and (4) estrous females selected traps containing odors from adult males but avoided those that had previously contained either nonestrous or pregnant/lactating females significantly more than expected. These findings have potential implications with regard to both the methods used for trapping small rodents and the social biology of house mice.     

Responses of beaver (Castor canadensis Kuhl) to predator chemicals

Free-ranging beaver (Castor canadensis) in two different beaver populations in New York State were exposed to predator chemicals to test feeding inhibition. Solvent extracts of feces were applied to stem sections of aspen, the preferred food tree of beavers, permitting smelling and tasting the samples. Predator odors were from wolf (Canis lupus), coyote (Canis latrans), dog (Canis familiaris), black bear (Ursus americanus), river otter (Lutra canadensis), lynx (Lynx canadensis), and African lion (Panthera leo). The experiment was repeated. The predator odors reduced feeding compared to untreated or solvent-treated controls. One population consumed 17.0% of the samples with predator odor and 27.0% of the controls in summer, and 48.4% and 60.0%, respectively, in autumn. The other population accepted 3.15% of the predator odor samples and 11.05% of the controls in summer. Coyote, lynx, and river otter odors had the strongest effects. Diesel oil and bitter-tasting neem extract had weaker effects. Predator odors are promising as feeding repellents for beaver.     

Δ22-β-Muricholic acid in monoassociated rats and conventional ratsacid in monoassociated rats and conventional rats

Bile acids were analyzed in the bile, small and large intestines, and feces of germ-free rats after a single inoculation with one of six intestinal bacteria that had been originally isolated from human feces.Bacteroides vulgatus andBifidobacterium longum preferentially deconjugated tauro-β-muricholic acid and taurocholic acid, respectively.Clostridium ramosum, Peptostreptococcus productus andLactobacillus gasseri deconjugated both bile acids, butEscherichia coli did not deconjugate either one. Rats inoculated with bacteria that deconjugated tauro-β-muricholic acid produced Δ²²-β-Muricholic acid in the feces. In contrast, Δ²²-cholic acid could not be detected in rats inoculated with bacteria that deconjugated taurocholic acid.     

Transformation of bile acids and sterols by clostridia (fusiform bacteria) in Wistar rats

The effects on bile acid and sterol transformation of clostridia (fusiform bacteria), the dominant intestinal bacteria in rodents (ca. 10¹⁰ counts per g wet feces) were examined in Wistar rats. After inoculation of clostridia into germ-free rats and into rats previously inoculated solely with Escherichia coli, most of the endogenous bile acids were deconjugated, and cholic acid and chenodeoxycholic acid were 7α-dehydroxylated to deoxycholic acid and lithocholic acid, respectively. Tauro-β-muricholic acid, another major bile acid in rats, was deconjugated, but only part of it (ca. 30%) was transformed into hyodeoxycholic acid. Cholesterol and sitosterol were also reduced to coprostanol and sitostanol, respectively. Escherichia coli transformed neither bile acids nor sterols. These data suggest that clostridia play an imporant role in the formation of secondary bile acids and coprostanol in rats.