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.     

Phenolic compounds from male castoreum of the North American beaver,Castor canadensis

North American beaver (Castor canadensis) mark their territories with castoreum, the contents of their castor sacs. In their territories, beaver respond with scent marking to experimental scent marks consisting of castoreum, or selected single components. In part, the unique odor of castoreum is due to large amounts of phenolic compounds. Purified phenolic components were analyzed by GC, GC-MS, and NMR; identifications were confirmed by comparing the spectra of synthetic phenols with those of the isolated phenols. Of the 15 phenols reported elsewhere, only five were confirmed in our analysis; the other 10 phenolic compounds are either absent or are not volatile enough to be detected by our methods. In addition, 10 phenolic compounds have been identified in this study that were not reported in the previous papers concerning the constitution of castoreum.     

Beaver (Castor canadensis) responses to major phenolic and neutral compounds in castoreum

North American beaver (Castor canadensis) mark their territories with castoreum, a chemically complex secretion from their castor sacs. The phenolic and neutral fractions of castoreum have been shown to elicit specific behavioral responses from beavers in a field setting. Our objective was to identify compounds/mixtures that evoked responses similar to those stimulated by castoreum. We assayed recently identified phenolic compounds, some phenolics that had been determined to be biologically active in previous studies, the neutral compound borneol, and combinations of phenolic compounds, neutral compounds, and the two combined. Biological activity was measured by the elicitation and extent of specific responses and their strength (duration, frequency, and proportion of beavers responding). Generally, single compounds stimulated fewer responses than mixtures. A 26-compound mixture of phenolic and neutral compounds elicited responses in a similar proportion of trials as castoreum. However, responses to castoreum were stronger than to any synthetic sample. Further investigation of different measures of response, namely, elicitation, completeness, and strength, are deemed necessary to fully decipher the design of social odors.     

Bioactivity of beaver castoreum constituents using principal components analysis

North American beaver (Castor canadensis) were observed to sniff from the water and make land visits to some synthetic chemical components of castoreum placed on experimental scent mounds (ESM). In previous analysis, the elicitation (presence/absence), completeness, and/or strength (number, duration) of these key responses served as separate measures of biological activity. In this paper, we used principal components analysis (PCA) to combine linearly six related measures of observed response and one index of overnight visitation calculated over all trials. The first principal component accounted for a majority of the variation and allowed ranking of the samples based on their composite bioactivity. A second PCA, based only on response trials (excluding trials with no responses), showed that responses to the synthetic samples, once elicited, did not vary greatly in completeness or strength. None of the samples evoked responses as complete or strong as the castoreum control. Castoreum also elicited more multiple land visits (repeated visits to the ESM by the same individual or by more than one family member) than the synthetic samples, indicating that an understanding of the castoreum chemosignal requires consideration of responses by the family unit, and not just the land visit by the initial responder.     

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.     

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.     

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.     

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.     

Experimental habitat scenting inhibits colonization by beaver,Castor canadensis

Unoccupied beaver (Castor canadensis) sites in New York State were for two years experimentally scented with a mixture of beaver castoreum and anal gland secretion. These sites were colonized less often than unscented control sites. The beaver is the first mammal to have been shown experimentally to use intraspecific odor cues when settling in vacant habitat. Territorial pheromones may be useful as repellents for beaver or other rodents.     

The social role of scent marking in beaver (Castor Canadensis)

Beaver colonies with close neighbors constructed more scent mounds than did isolated colonies, and the number of scent mounds at each active lodge is correlated with the distance to the nearest occupied lodge. The scent mounds were typically located at trails, also on lodges and dams. The beaver rebuilt experimentally removed scent mounds. Experimentally scent-marked unoccupied lodges were less often visited or inhabited than unscented control lodges. Resident beaver responded to artificial scent marks near their lodges with aggressive behavior and increased activity. We conclude that scent mounds serve in delineating family territories and are effective in deterring transient beaver from utilizing existing but uninhabited lodges.     

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.     

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.     

Floral Odors of Silene otites: Their Variability and Attractiveness to Mosquitoes

Inflorescence scent samples from nine populations of dioecious Silene otites, a plant pollinated by moths and mosquitoes, were collected by dynamic headspace extraction. Sixty-three scent samples were analyzed by gas chromatography–mass spectrometry. Out of 38 found, 35 compounds were identified, most of which were monoterpenoids, fatty acid derivatives, and benzenoids. Phenyl acetaldehyde was the most dominant compound in the majority of samples. The variability in scent composition was high, and population and sex differences were found. Nevertheless, wind tunnel experiments proved similar attraction of Culex pipiens pipiens biotype molestus mosquitoes to the inflorescence odor of S. otites of different populations, indicating that different blends are similarly attractive to mosquitoes. The electrophysiological responses of mosquitoes to the 12 most common and abundant odor compounds of S. otites differed. Linalool oxide (furanoid) and linalool evoked the strongest responses in male and female mosquitoes, and (Z)-3-hexenyl acetate was strongly active in females. Medium responses were evoked in males by (Z)-3-hexenyl acetate, in females by benzaldehyde and methyl salicylate, and in both sexes by lilac aldehyde, lilac alcohol, and linalool oxide (pyranoid).     

Antennal Responses to Floral Scents in the Butterfly Heliconius melpomene

Floral scent, together with visual floral cues, are important signals to adult butterflies searching for food-rewarding plants. To identify which compounds in a floral scent are more attractive and, thus, of biological importance to foraging butterflies, we applied electrophysiological methods. Antennal responses of male and female adults of the tropical butterfly Heliconius melpomene L. (Lepidoptera: Nymphalidae: Heliconiinae) to individual compounds of natural floral scents and synthetic floral scent mixtures were investigated using gas chromatography–electroantennographic detection (GC-EAD). The natural floral scents included those of two tropical plant species, Lantana camaraL. (Verbenaceae) and Warszewiczia coccinea (Vahl) Kl. (Rubiaceae), and two temperate species, Buddleja davidii Franchet (Loganiaceae) and Cirsium arvense (L.) Scop. (Asteraceae). The two synthetic floral scent mixtures contained many of the compounds found in the natural scents, but all in equal quantities. Compounds both present in relatively high abundance in the floral scents and detected exclusively in the floral parts of the plant, such as linalool, linalool oxide (furanoid) I and II, oxoisophoroneoxide, and phenylacetaldehyde, elicited the strongest antennal responses, suggesting that they may reflect adaptations by the plant to attract butterfly pollinators. However, other compounds also present in high abundance in the floral scent, but detected in the vegetative as well as floral plant parts, either elicited strong antennal responses, such as trans--ocimene and benzaldehyde, or failed to elicit antennal responses, such as the sesquiterpenes -caryophyllene and -humulene from L. camara. The most volatile monoterpene alkenes in the synthetic scent mixtures elicited only low or no responses. Furthermore, the overall antennal responses were stronger in females than in males. The findings suggest that several floral scent volatiles, especially those of exclusively floral origin, are of high biological significance to H. melpomene butterflies. These include compounds of different biosynthetic origins belonging to the benzenoids, monoterpenoids, and irregular terpenoids.     

Phenolic compounds isolated from bitter lupine seeds and their inhibitory effects on germination and seedling growth of lettuce

The phenolic acids, including 4-hydroxybenzoic and 4-hydroxycinnamic acids, and their derivatives, such as 6,7-dihydroxycoumarin and 1,2-dihydroxybenzene, were isolated from bitter lupine seeds and were identified using gas chromatography-mass spectrometry. These compounds inhibited lettuce seed germination in the first 24 hr after sowing, but after 72 hr germination was comparable with that of the control. However, very strong suppression of seedling growth, especially the roots, was observed for higher concentrations of the lupine seed fractions containing phenolic acids. Effects observed in the lettuce germination bioassays were compared with those produced by nine pure phenolic acids previously identified in the mixture. The ethyl acetate fraction from lupine seed extract inhibited seedling growth as effectively as pure 1,2-dihydroxybenzene, the strongest inhibitor of the pure phenolic compounds studied. The possible reason for this could be the synergistic effect created in the mixture of phenolic compounds isolated from the extract.     

Identification of Floral Volatiles From Ligustrum japonicum that Stimulate Flower-Visiting by Cabbage Butterfly, Pieris rapae

Floral scent compounds of Ligustrum japonicum that affect the foraging behavior of Pieris rapae adults were examined by means of chemical analyses, electroantennogram (EAG) responses, and behavioral bioassays; the behavioral biossays consisted of two tests: reflex extension of proboscis (REP) in response to odor, and attraction to scented and unscented artificial flowers. More than 30 compounds, including 2-phenylethanol, benzyl alcohol, and methyl phenylacetate as the major components were identified from L. japonicum flowers. Of these, 22 compounds were tested for their effect on foraging behavior. Phenylacetaldehyde (PA), 2-phenylethanol (PE), and 6-methylhept-5-en-2-one (MHO) elicited the highest REP responses, and benzaldehyde (BA) and methyl phenylacetate (MPA) evoked intermediate REP responses. EAG responses were not necessarily correlated with REP activities; the three high-REP compounds gave only moderate EAG responses, whereas two other compounds (ethyl phenylacetate and 2-phenylethyl acetate) that released high EAG responses showed low REP activities. In two-choice behavioral bioassays, flower models scented with any one of these high-REP compounds attracted significantly more adults, while compounds with low REP activities exhibited weak or no appreciable attractiveness. This suggests that the REP responsiveness closely reflects the attractiveness of a compound and could be an effective measure in elucidating which chemical attractants are involved in flower-visiting. A synthetic blend of five floral chemicals (PA, PE, MHO, BA, and MPA) displayed an attractiveness that was comparable to that of the floral extract and was more effective in attractiveness than the compounds tested singly. Consequently, it is highly likely that the flower-visiting by P. rapae to L. japonicum is mediated largely by floral scent chemicals and that a synergistic effect of the five floral components would be most responsible for attraction of the butterfly to this flower. The present results also strongly suggest that specific floral volatiles may facilitate close-range flower location by P. rapae, could serve in part as a cue for recognizing food sources, and also be profoundly implicated in flower preference.     

1,4-Dimethoxybenzene,a Floral Scent Compound in Willows that Attracts an Oligolectic Bee

Many bees are oligolectic and collect pollen for their larvae only from one particular plant family or genus. Here, we identified flower scent compounds of two Salix species important for the attraction of the oligolectic bee Andrena vaga, which collects pollen only from Salix. Flower scent was collected by using dynamic-headspace methods from Salix caprea and S. atrocinerea, and the samples were subsequently analyzed by coupled gas chromatographic–electroantennographic detection (GC-EAD) to detect possible attractants of A. vaga. EAD active compounds were identified by gas chromatography coupled to mass spectrometry. Both Salix species had relatively similar scent profiles, and the antennae of male and female bees responded to at least 16 compounds, among them different benzenoids as well as oxygenated monoterpenoids and sesquiterpenoids. The strongest antennal responses were triggered by 1,4-dimethoxybenzene, and in field bioassays, this benzenoid attracted females of A. vaga at the beginning of its flight period, but not at the end.     

Flower Scent of Floral Oil-Producing Lysimachia punctata as Attractant for the Oil-Bee Macropis fulvipes

Most flowers offer nectar and/or pollen as a reward for pollinators. However, some plants are known to produce mostly fatty oil in the flowers, instead of nectar. This oil is exclusively collected by specialized oil-bees, the pollinators of the oil-plants. Little is known about chemical communication in this pollination system, especially how the bees find their hosts. We collected the floral and vegetative scent emitted by oil-producing Lysimachia punctata by dynamic headspace, and identified the compounds by gas chromatography coupled to mass spectrometry. Thirty-six compounds were detected in the scent samples, several of which were flower-specific. Pentane extracts of flowers and floral oil were tested on Macropis fulvipes in a biotest. Flower and oil extracts attracted the bees, and some of the compounds identified are seldom found in the floral scent of other plants; these may have been responsible for the attraction of the bees.     

Responses by domestic cats (Felis catus) to snake scent gland secretions

The scent gland secretions of snakes are thought to repel predators, but few predator species have been tested for responses to these exudates. Domestic cats (Felis catus) were tested for responses to scent gland secretions of the gray rat snake (Elaphe obsoleta), or to choloroform extracts of them, applied to filter paper or food. More cats salivated or rubbed on filter papers treated with scent gland secretions than on control papers. Scent gland exudates elicited rubbing and pawing in cats more frequently than did chemicals from a shed snake skin. Cats offered food pieces treated either with water or with scent gland secretions ate fewer of the latter; this result is consistent with the hypothesis that scent gland secretions deter feeding.     

Castor sacs and anal glands of the north american beaver (Castor canadensis): their histology,development, and relationship to scent communication

Both sexes of beavers possess a pair of castor sacs and a pair of anal glands located in paired subcutaneous cavities between the pelvis and the base of the tail. The castor sacs are not glandular in the histological sense, hence references to these structures as preputial glands or castor glands are misnomers. The wall of the castor sacs is plicate and comprised of three distinct zones: an outer layer of vascular connective tissue, a two-to five-cell-thick layer of mitotic epithelial cells, and several densely packed layers of cornified epithelium which grade into more widely separated sheets toward the lumen. Monocultures of a gram-positive facultatively anaerobic bacterium were present in the lumen of all castor sac preparations. Differences in the frequency of castoreum deposition were not attributable to differences in the structure of the castor sacs. The anal glands of beavers are holocrine sebaceous glands. These glands develop more rapidly than the castor sacs. Anal gland tissue from embryos exhibited cellular characteristics associated with the production of sebum. Secretory activity was evident in all preparations. The relationship of castoreum and anal gland secretion to scent communication among beavers is discussed.