Odorant Source Used in Eurasian Beaver Territory Marking

Mammals use urine, feces, or the secretion of specialized skin glands to mark their territories. These sources can carry different information and, thus, have different functions. Presently it is not known if beavers (Castor spp.) deposit castoreum (primarily a mixture of secondary metabolites from urine) from the castor sacs and secretion from the anal glands (AGS) together or alone when scent marking their territories. We hypothesized that castoreum would be the main scent signal used in the defense of beaver territories during winter and predicted that castoreum would be deposited more often than AGS. A total of 96 scent marks on snow were collected from January 1 to March 31, 1997–1999 in the Bø River, Telemark County, Norway. In order to obtain control material, we chemically analyzed AGS and castoreum from 60 dead beavers collected during January–May 1997–1999. We compared the compounds found in the dead beavers with compounds found in the scent marks on snow. Samples were analyzed by using gas chromatography–mass spectrometry (GC-MS). All 96 scent marks contained compounds from castoreum, whereas compounds from AGS were found in only four scent marks. This suggests that beavers do not specifically deposit AGS on scent mounds as they do with castoreum and that the AGS compounds we found probably were remnants of AGS from the feet or fur following pelt lubrication or coprophagy behavior. We conclude that castoreum is the main scent signal used in the defense of beaver territories during winter.     

Scent-Marking in the Eurasian Beaver (Castor fiber) as a Means of Territory Defense

Beaver (Castor spp.) normally scent-mark by depositing castoreum and/or anal gland secretion on scent mounds close to the water's edge. The aim of this study was to investigate the hypothesis that the Eurasian beaver (C. fiber) scent-marks as a means of territory defense. Scent-marking behavior was studied during an entire year (April 1, 1995–March 31, 1996) in seven adjacent territories along 9.2 km of the B River in Telemark County, Norway. The number and location of fresh scent marks were recorded biweekly. The main results showed that: (1) the number of scent marks in territories was significantly higher in spring, when dispersal of subadults normally occurs than during the rest of the year; (2) the number of scent marks was clumped near territorial borders; and (3) the number of scent marks was significantly greater upstream than downstream of the lodge.     

Increased scent marking in male Mongolian gerbils by urinary polypeptides of female conspecifics

Male Mongolian gerbils scent mark during a standardized openfield test on an individually different but more-or-less constant level in the absence of females. The presence of females in the same room but without direct contact to the males increases the marking frequencies of the males to an individually different higher level. A similar increase in marking frequency is obtained by application of female urine directly to the nostrils of the males, but not by male gerbil urine or female laboratory mouse urine. This indicates that substances present in female gerbil urine are responsible for the increase in male marking behavior. Using this bioassay, various fractions of female urine were tested for their influence on marking behavior. Our results to date show that the component increasing male scent-marking behavior is hydrophilic and nonvolatile and is contained in the nondialyzable, high-molecular-weight polypeptide fraction of female gerbil urine.     

Social Signals Involved in Scent-Marking Behavior by Cheek-Rubbing in Alpine Marmots (Marmota marmota)

The Alpine marmot Marmota marmota is a territorial rodent. Resident adults regularly scent-mark their territory by cheek-rubbing, mainly on burrow entrances and along boundaries. The purpose of this three part study was to gain further insight into this scent-marking behavior by: (1) observing the response of free-ranging marmots to foreign scent marks, (2) confirming the glandular source of the marking substance by histologic examination of the temporal gland, and (3) identifying biologically active chemical fractions of the marking substance. To allow field tests, we developed a device consisting of a glass tube placed upside down over a stake. Two devices were simultaneously placed at one burrow entrance. On one device, a clean tube was used and, on the other, a tube alternatively coated with either whole natural scent-marking substances or various fractions obtained by solvent extraction or chromatographic separation from whole scent-marking substances. Subsequent observations showed a significant difference in the duration of nose contact and number of cheek-rubbing movements. Resident adult marmots sniffed and marked tubes bearing alien marks significantly more than clean control tubes. Similar differences in behavior were observed with ethanol extracts of whole scent-marking substances. Extracts obtained with pentane and dichloromethane showed no bioactivity, suggesting that highly polar compounds are the active substances in the Alpine marmot. The temporal gland is an exocrine gland located on each side of the head with numerous pores opening at the surface of the skin in the cheek area. GC-MS analysis of individually collected samples from these glands showed that over 30 compounds were consistently present. Seven of these compounds were identified. Two fractions were obtained and used together and separately in field tests. Fraction 1 was composed mainly of short-chain alcohols and alkanes, and fraction 2 had a more acid and ester composition. The fact that these two fractions were active together but not separately strongly suggests that the active territorial signal results from a synergistic interaction between several compounds.     

Some communicatory functions of scent marking in the cotton-top tamarin (Saguinus oedipus oedipus)

The ability of cotton-top tamarins to discriminate between scents from conspecifics and those from other tamarin species, and between scents from conspecific individuals was tested. Cotton-tops scent mark with specialized skin glands in the circumgenital area. Females possess larger glands than males and show more scent-marking behavior. In the first experiment, subjects were presented with a glass rod scented with either material collected from the surface of the scent glands of a conspecific female, with scent material from a female of a related species, or with an unscented rod. Glass rods carrying scent from conspecifics were sniffed more frequently than rods carrying scent from related tamarin species or unscented rods. A second experiment offered a choice between two glass rods, one scent marked by aSaguinus o. oedipus female, the second one scent marked by aSaguinus fusdcollis female. Shelves carrying rods that had been scent marked by conspecifics were contacted more frequently than those carrying rods marked by heterospecific females. Scent marks from conspecific females were also sniffed more frequently. A third experiment compared the level of responses to rods carrying material collected from the scent glands of female individuals to which the subjects had been habituated with their responses to rods carrying scent from females to which they had not been habituated. Contacting and sniffing responses to the scents of novel females were higher than those to the scents of females to which the subjects had been habituated.     

Do Eurasian beavers smear their pelage with castoreum and anal gland secretion?

The scent-matching hypothesis postulates that scent marks provide an olfactory link between a resident owner and his territory, and that this enables intruding animals to recognize the chance of escalated conflicts. However, it is unclear if Eurasian beavers (Castor fiber) mark their own pelage with castoreum from their castor sacs (i.e., the same material used in territorial marking); and/or if beavers mark their pelage with anal gland secretion (AGS) from the anal glands to waterproof the pelage and to act as a living-scent mark. Chemical analysis (gas chromatography and mass spectrometry) of hair samples from 22 live-trapped beavers revealed that castoreum compounds were not present in any samples, AGS compounds were found from 3 animals (13.6%) around the cloaca, and the compound squalene was found in all the samples. Beavers may release castoreum directly into the water when it meets an intruder. Thereby, the scent mark in the water can provide an olfactory link between a resident owner and his territory. Squalene, in contrast to AGS, may be essential for keeping beaver pelts water-repellant.     

Chemical Characterization of Urinary Pheromones in Brown Antechinus, Antechinus stuartii

In the small dasyurid marsupial, Antechinus stuartii, males exhibit scent-marking in the form of cloacal marking of nesting areas during the breeding season. Females of this species show no such behavior. To characterize the potential male pheromonal scent signal, urine-derived volatiles from sexually active males were analyzed by GC-MS and compared to that of females and a castrated male. More than 10 urinary compounds were identified. A series of homologous methylketones was observed in both males and females, whereas aldehydes were present only in female urine. Urine from the castrate was virtually compound-free except for minute concentrations of a compound tentatively identified as 2,4-dithiapentane. This compound was also found in one of the sexually active males. The GC profiles of the sexually active males contained high concentrations of two pyrazine derivatives and four methylketones that were not detected in the profiles of either females or the castrate. These compounds may influence social communication in the brown antechinus, Antechinus stuartii.     

Factors Affecting Scent-Marking Behavior in Eurasian Beaver (Castor fiber)

We tested the hypothesis that a main function of territory marking in Eurasian beaver (Castor fiber) is defense of the territory. The results showed that: (1) beaver colonies with close neighbors scent-mark more often than isolated ones; (2) the number of scent markings increased significantly with the number of neighboring territories and individuals, the mean distance to all other territories, duration of territory occupancy and length of wooded banks within the territory; (3) the number of scent markings by a colony was not related to the number of animals in the colony, the distance to nearest neighbors' main resting-site, or age of beaver; (4) the number of scent mounds or scent markings in territories with breeding and nonbreeding beaver did not differ; (5) there was no significant difference in the number of scent markings during the season (16 April–31 August); and (6) some of the scent mounds were concentrated at feeding sites, resting sites, and near trails, but most had no apparent relationship to any of these sites. Therefore we concluded that scent marking apparently plays an important role in territory defense of the Eurasian beaver.     

Neutral compounds from male castoreum of North American beaver,Castor canadensis

North American beavers (Castor canadensis) mark their territories with castoreum, the strong-smelling paste in their castor sacs. In their own territories, beavers respond with scent marking to experimental scent marks that consist of strange castoreum (or selected components). In part, the unique odor of castoreum is due to large amounts of phenolic compounds and neutral compounds. Purified neutral compounds were analyzed by GC. GC-MS, and NMR; identities of the neutral compounds were confirmed by comparing the properties of authentic compounds with those of the isolated compounds. We identified 13 neutral compounds that had not been reported before for castoreum. Most of these are oxygen-containing monoterpenes. Of the nine neutral compounds reported by Lederer (1949), only three are confirmed in our analysis; the other six neutral compounds are either absent or are not volatile enough to be detected by our methods. Eight compounds—6-methyl-l-heptanol, 4,6-dimethyl-l-heptanol, isopinocamphone, pinocamphone, two linalool oxides, and their acetates—were synthesized for structure identification and bioassays.     

Detection and Discrimination of Conspecific Scents by the Anguid Slow-Worm Anguis fragilis

We tested the ability of male slow-worms, Anguis fragilis, a limbless anguid lizard with secretive, semifossorial habits, to detect chemical associated with conspecifics by using a T-maze in the laboratory. Male slow-worms discriminated conspecific male and female scent deposits. Males selected the arm with female scent, suggesting that scent deposits may be used to locate potential mates. Also, male slow-worms did not avoid the chemicals of other males, suggesting that they are not territorial. However, males discriminated their own scent from those of other males, and spent more time exploring the arm with other male scent, which suggests that scent marks may bear information that could be used in future intrasexual social contexts. We conclude that discrimination of conspecifics based on scents may be more widespread than previously expected among lizards inhabiting visually restricted environments.     

Scent marks as social signals inGalago crassicaudatus I. Sex and reproductive status as factors in signals and responses

In simultaneous presentations of the scent marks of male and female conspecifics, thick-tailed galagos (Galago crassicaudatus) of both sexes responded most strongly to the female scent. Males differentiated between the scents most clearly in their sniffing, females in their scent marking in response. The scent of estrus females elicited increased licking of the scent by both sexes. This greater response to female scent and the contrasts in response patterns between the sexes are discussed and related to galago social and spatial relationships in the wild.     

Scent trailing by virgin females ofPseudococcm calceolariae

Virgin females of the citrophilous mealybugPseudococcus calceolariae (Mask.) deposit scent marks as trails on the substrate on which they rest or move. These substances elicit attraction and sexual behavior by conspecific males. The same responses were obtained when males were bioassayed on extracts from filter paper disks on which females had rested. The significance of scent trailing in mealybugs is discussed.Homoptera, Coccoidea, Pseudococcidae.This work was supported by the Italian National Research Council (CNR).     

Partial chemical characterization of urinary signaling pheromone in tree shrews (Tupaia belangen)

Tree shrews of both sexes exhibit marking behavior (chinning) in response to scent marks made by the urine of fertile male conspecifics. To isolate the effective odor components, the urine was fractionated by liquid-liquid extraction and TLC, and the fractions were tested by bioassay. The results show that chinning is elicited by several lipophilic urine fractions, which are more effective in combination than alone. To characterize the complex scent signal, the lipophilic extracts from urine of the two sexes were analyzed by GC-MS and compared. The GC profile of the males is distinguished by pyrazine compounds not detected in the profile of the females. The profiles of the sexes also differ with regard to several volatile monocarboxylic acids, which are present at higher concentrations in male than in female urine. More than 30 urine components have been identified. Synthetic equivalents of these urine components were bioassayed for effectiveness in eliciting chinning and compared with one another as well as with scent substances not normally present in tree-shrew urine. Strong chinning responses were elicited by (1) certain pyrazine compounds and (2) some monocarboxylic acids, when presented at the high concentrations specific to male urine. Marking behavior is usually not elicited by scent substances not contained in tree-shrew urine or by urine components common to mammals in general. The data so far available indicate that the male-specific scent signal of tree shrews is based less on a single unique component than on the qualitative and quantitative characteristics of a multicomponent mixture. In the bioassay, tree shrews of both sexes respond equally to the male specific substances by chinning. As the scent signal represented by these substances has a different meaning to males and females (rival or potential mate, respectively), chinning probably serves several different functions.     

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.     

Volatile components of scent material from cotton-top tamarin (Saguinus o. oedipus)

The first chemical analysis of the volatile components of scent material from the cotton-top tamarin,Saguinus o. oedipus is presented. In contrast to our previous findings in saddle-back tamarins (Saguinus fuscicollis), the chemical composition of the scent marks is quite variable. Analysis of secretion expressed manually from different areas of the scent pads of sedated animals shows individual and regional variability. Only three components (squalene, cholesterol, andp-methoxybenzaldehyde) were found in all scent mark samples analyzed. However, suprapubic secretions obtained from one sedated female contained, in addition, 12 butyrate esters as well as five acetate esters which were homologous to the butyrates. The butyrate esters have been previously identified in scent material fromSaguinus fuscicollis. The variability of the composition of the scent material is discussed in relation to the scent-marking behavior of the cotton-top tamarins. Behavioral studies tested the ability of the cotton-top tamarins to discriminate between scent marks from conspecifics and scent marks from saddle-back tamarins. The group of 12 subjects discriminated between the scent marks from both species in choice tests during which they could freely contact the scent samples. However, when scent marks were presented under a screen, so that the subjects could smell but not contact the stimuli, no discrimination was shown. Four individuals displayed more interest in the screened scent stimuli than all other subjects. When these animals were tested in a second experiment for their ability to discriminate between material from the two species on the basis of only volatile cues, it became evident that they were able to do so. These results indicate that volatile cues alone enable the tamarins to recognize scent marks from conspecifics, but that additional cues perceived during contact with the scent are important for its full attractiveness and/or informational content.     

Physiological Stress Responses,Fecal Marking Behavior,and Reproduction in Wild European Pine Martens (<Emphasis Type="Italic">Martes martes</Emphasis>)

The relationship among physiological stress responses, fecal marking behavior, and reproduction in male and female European pine martens was investigated. Between July 2004 and June 2007, 145 fresh fecal samples were collected in a protected area of northwest Spain. Fecal DNA was used for specific identification by using the polymerase chain reaction-restriction fragment length polymorphism technique. Glucocorticoids (cortisol) and sex steroid hormones (P, progesterone; E, estradiol; T, testosterone) were quantified by enzyme immunoassays. Sex was assigned according to concentrations of T+P+E and the T/P ratio. Fecal cortisol concentrations were higher in males than in females. Feces with a presumptive marking function (on conspicuous substrates, above ground level, and/or in latrines) had higher mean levels of cortisol than those that were on inconspicuous substrates and/or at ground level, for both males and females. Fecal mark density was highest in spring, when mean levels of fecal cortisol were more elevated. Therefore, the higher physiological stress levels in females could be due to female physiological state (late-term pregnancy and lactation), competition for resources connected to birth, or food resources for offspring rearing. In males, the increase could be due to higher male competition for access to females during pro-estrus and estrus. Our results suggest that scent marking in European pine martens is related to reproduction and is involved in intersexual and intrasexual communication.     

Analysis of chemical signals in a primate species (Saguinus fuscicollis): Use of behavioral,chemical, and pattern recognition methods

Scent marking with specialized skin glands is a common behavior in the tamarin,Saguinus fuscicollis. The scent marks identify species, subspecies, gender, and individual, and they also contain information on the social position and hormonal condition of an animal. The marks are chemically complex, containing a large number of compounds. Analysis by means of gas chromatography-mass spectrometry has identified 16 major components (squalene and 15 esters of butyric acid). These compounds are present in the marks of males and females of two subspecies,Saguinus f. fuscicollis andSaguinus f. illigeri. Application of computerized pattern recognition techniques has shown that concentration patterns of some of the butyrates are diagnostic of the two subspecies while concentration patterns of other butyrates are diagnostic of males and females regardless of subspecies. Behavioral studies have shown that the concentration patterns of butyrates and squalene alone do not encode information on subspecies and gender. It is, however, likely that this information is partially encoded by these specific butyrate-squalene concentration patterns but that yet unidentified compounds in the scent marks serve as necessary synergists.     

Differential Display of Investigative Behavior Permits Discrimination of Scent Signatures from Familiar and Unfamiliar Socially Dominant Female Marmoset Monkeys (Callithrix jacchus)

The hypothesis was tested that marmosets have differential displays of investigatory behavior towards circumgenital scent marks from socially dominant females based on degree of familiarity. In a series of two-choice behavioral discrimination bioassays, dominant males together with subordinate and dominant females were presented concurrently with scent secretions from familiar versus unfamiliar dominant females, and their investigative behaviors towards the scent were recorded. Test animals directed significantly different amounts of investigative behaviors towards familiar versus unfamiliar scents when the scent stimuli involved complete circumgenital scent marks, glandular secretion only, or urine only. When animals discriminated between the two scent samples, they did so by directing significantly increased amounts of behavior to scent from unfamiliar females. Chemical cues promoting differential displays of behavior to familiar versus unfamiliar scents appeared to be volatile in nature and were effective in the scent mark for at least one day following deposition. The reliable ability of marmosets to consistently discriminate between odors from familiar versus unfamiliar dominant females provides strong circumstantial evidence that individual female marmosets have distinct chemical signatures. The ability of marmosets to discriminate the odor from familiar versus unfamiliar dominant females may be important in the context of intergroup relations.     

Scent Marking in Male Premating Behavior of Bombus confusus

Premating behavior in the species Bombus confusus Schenck, 1859 was studied. The marking habits of bumblebee males were revealed and the use of their labial gland secretion in scent marks was proved chemically. Identical compounds were present both in the labial gland secretion and on male-marked objects. This chemical proof is reported for the first time from natural conditions of a B. confusus locality. Males' flight activities, site tenacity, and male–male interactions are described in detail. Contrary to literature reports, male B. confusus behavior does not basically differ from that of already known perching bumblebee species.     

Variation of Floral Scent Emission and Postpollination Changes in Individual Flowers of Ophrys sphegodes Subsp. sphegodes

We investigated the scent composition of individual flowers of Ophrys sphegodes, its alteration following pollination, and of picked flowers by day and at night. Odor samples were collected by headspace sorption and analyzed by gas chromatography and mass spectrometry. To evaluate the function of postpollination odor changes, we carried out behavioral tests on the pollinator Andrena nigroaenea with pollinated and unpollinated flowers. We identified 27 volatiles in the flower scents. Aldehydes and alkanes were most frequently found. Aldehydes were the most abundant class of compounds (40–50%). When flowers were picked, they emitted significantly lower total amounts of volatiles than unpicked flowers, and their odor bouquets were significantly different. Comparison of scents released by day and at night showed no decrease in scent emission during nighttime, but the odor bouquets were significantly different. Pollinated flowers produced significantly different odor bouquets, and the total amount of scent emitted two to four days after pollination was significantly lower compared with unpollinated flowers. In addition, behavioral tests with A. nigroaenea males showed that flowers were significantly less attractive three days after pollination. This reduced attractiveness is hypothesized to guide pollinators to the unpollinated flowers within an inflorescence, and thus increase the reproductive success of the plant.