Misidentification by wild rabbits,Oryctolagus cuniculus,of group members carrying the odor of foreign inguinal gland secretion

The results of experiments designed to clarify the behavioral function of the odor of the secretion from the inguinal glands of rabbits,Oryctolagus cuniculus, are presented. Members of groups of female rabbits were smeared with odoriferous materials, and the behavior of their penmates towards them was recorded. Eight sources of natural rabbit odors and a commercial perfume were used in 180 tests involving 120 adult female wild-type rabbits. A high proportion of the animals smeared with the inguinal gland secretions from unfamiliar females were attacked by their pen-mates. In contrast, treatment with inguinal gland secretions from unfamiliar male rabbits; macerated inguinal gland materials from unfamiliar males and females, respectively; urine from unfamiliar females; and macerated anal gland material from unfamiliar females resulted in only minor disruption to the normal social relationships between the group members which was not significantly different to that caused by treatment with a commercial perfume.     

Misidentification by wild rabbits,Oryctolagus cuniculus,of group members carrying the odor of foreign inguinal gland secretion

To demonstrate that the inguinal gland odor conveys information on the identity of individuals, 60 male, wild-type rabbits,Oryctolagus cuniculus, living in groups of three in outdoor pens, were used in 120 tests. Members of the groups smeared with the inguinal gland secretion from unfamiliar rabbits were attacked by their penmates. In contrast the odors of chin gland secretions and urine of strange rabbits and of a commercial perfume did not cause serious disruption to the social relationships within the groups.     

Effect of volatiles collected above fecal pellets on behavior of the rabbit,Oryctolagus cuniculus,tested in an experimental chamber

The fecal pellets of rabbits,Oryctolagus cuniculus, are coated with secretion from the anal gland, the odor from which functions as a territorial marker. The total volatiles collected from above fecal pellets and three different fractions prepared by washing the volatiles through either distilled water, 1 M sodium hydroxide, or 1 M hydrochloric acid were tested for their effect on the territorial confidence of individual animals. Tests were based on the neutral pen paradigm used in previous studies but employed a specially designed test chamber, the internal odor environment of which could be fully controlled. In the four series of bioassays a total of 140 separate tests were performed and 64 adult male rabbits were used. The results showed that the presence of the total volatiles influences the outcome of territorial competitions in pairs of male rabbits in favor of the individual from which they were derived. They were comparable with those obtained in earlier studies when fecal pellets were presented in the test pen. The effects of the different washing reagents on the total odor are illustrated by gas chromatograms, and the components most likely to be removed by the different treatments are suggested. The volatiles remaining after passing through distilled water had less effect on the rabbit's behavior than the total odor. Since the washing process not only removed water-soluble compounds but also led to a reduction in the concentrations of water-insoluble components introduced into the test chamber, either of these factors could have been responsible for the weakened effect. Fractions of the odor obtained by washing with either sodium hydroxide or hydrochloric acid did not influence the results of the tests. This suggests that the essential components of the olfactory signal include both basic and acidic compounds.     

Effect of volatiles collected above fecal pellets on behavior of the rabbit,Oryctolagus cuniculus,tested in an experimental chamber

Gas chromatographic techniques were used to trap and fractionate the volatile substances emanating from the fecal pellets of wild rabbits,Oryctolagus cuniculus, which incorporate the odor of the anal gland secretion. The volatiles were bubbled through water, trapped on Chromosorb 105, transferred to Silicone SF96 traps, passed through a GC capillary column, and retrapped on SF96. The behavioral effect of trapped and fractionated volatiles was determined in four series of bioassays involving 51 adult, male, wild-type rabbits in 112, ten-minute tests. The bioassay was based on the demonstration of territorial confidence by the rabbits under the influence of their own odor. In the bioassays of the unfractionated volatiles eluted from Chromosorb 105 and SF96 traps the animals were significantly more confident in the presence of the volatiles from their own fecal pellets. One of the two fractions of the total volatiles was ineffective while the effect of the other was less than that of the unfractionated odor. The results demonstrate that the gas chromatographic techniques can be used to manipulate complex mammalian odors with precision. Combined with a discriminative bioassay, this opens up the possibility of identifying the specific combinations of volatile substances involved in the formation of olfactory signals.     

Misidentification by wild rabbits,Oryctolagus cuniculus,of group members carrying the odor of foreign inguinal gland secretion

The experiments described in this paper are part of a series designed to clarify the behavioral function of the odor of the secretion from the inguinal glands of rabbits,Oryctolagus cuniculus. Results are presented of tests carried out on 48 mixed-sex groups consisting of 162 adult wild-type rabbits, 48 males and 114 females. The subordinate females of the groups were smeared with various odorous materials and the reactions of their pen-mates towards them recorded. Eight sources of natural rabbit odors and a commercial perfume were used in 198 tests. A high proportion (60%) of the subordinate females smeared with the inguinal gland secretions from unfamiliar males were attacked, mainly by the males, which were generally the most dominant individuals within the groups. The paper also presents statistical analyses of behavioral data collected during two earlier related experiments with single-sex groups involving 258 rabbits in 300 tests which have not previously been evaluated. The results throughout the whole study indicate clearly that the odor of the inguinal gland secretion of rabbits carries information which affects the acceptance of individuals by their companions in organized groups. Other sources of unfamiliar rabbit odor did not influence the attitude of groupmates toward the individuals smeared with them, or if they did, their effects were only marginal.     

Identification of some volatile compounds in the odor of fecal pellets of the rabbit,Oryctolagus cuniculus

A study has been made of the volatile constituents in the vapors collected from fecal pellets of wild rabbits,Oryctolagus cuniculus. Measurements of changes in the heart rates of adult male rabbits exposed to the effluent of a gas chromatographic capillary column were used to indicate the presence of compounds which may be of behavioral significance. Combined with the use of high-resolution columns in gas chromatography and gas chromatography-mass spectrometry, this approach has implicated several classes of compounds, including volatile fatty acids and phenols, which in certain absolute or relative concentrations may be involved in the formation of signals of territorial importance.     

Microflora and volatile fatty acids present in inguinal pouches of the wild rabbit,Oryctolagus cuniculus,in Australia

Contents of inguinal pouches of the wild rabbit,Oryctolagus cuniculus, have been examined for the presence of microorganisms and volatile fatty acids. A total of 245 isolates were made from 167 samples taken from 75 rabbits, and microorganisms were obtained from 93% of individuals. The most common microorganism encountered wasStaphylococcus aureus, followed by the yeastCandida kruzei. Other organisms isolated wereBacillus subtilis,Escherichia coli, andStreptococcus faecalis. Quantitative analysis of volatile fatty acids in scrapings from pouches showed considerable variation between individuals. Of the acids identified, acetic acid and isovaleric acid were the most abundant. In a liquid medium containing one of these acids, the growth of microorganisms was only slightly affected. However, growth became inhibited when these fatty acids were used in combination.     

Quantitative analysis of the effects of ultrasound from an odor sprayer on moth flight behavior

A piezoelectric sprayer was recently developed for precision release of odor stimuli in olfactory research. The device replaces conventional dispensers used to release semiochemicals in studies of moth flight toward odor sources. However, the device generates high-frequency sounds in the range that some moths can hear. Ultrasound from the standard set-up sprayer had a considerable impact on flight behavior of the silver Y moth, Autographa gamma, tested in a flight tunnel. It was affected at all behavioral stages when the dispenser was driven at 120 kHz. Only 5% of the moths reached the source when exposed to 120-kHz sound from the dispenser compared to 65% in the control group without sound. The proportion taking flight was also reduced. Hearing threshold curves obtained electrophysiologically revealed that moths were sensitive to the frequency range at which the sprayer was operated and that sound intensity from the sprayer was up to 40 dB above the moths' electrophysiological hearing threshold. The audiogram for A. gamma was similar to audiograms obtained for other noctuids. Hearing sensitivity was highest at around 15 kHz, where the threshold was 35 dB SPL (sound pressure level). The threshold increased with frequency up to 94 dB SPL at 160 kHz. We improved the sprayer to operate at 300 kHz, which is beyond the hearing ability of most insects with ears. At this high frequency, the moths' sensitivity to ultrasound is reduced considerably, and we did not observe any effect on flight behavior compared to a control group without sound. Accordingly, this new piezoelectric sprayer can be used with ultrasound-sensitive insects and insensitive insects alike.     

To fix or not to fix: the role of 2-phenoxyethanol in rabbit,Oryctolagus cuniculus,chin gland secretion

The European rabbit (Oryctolagus cuniculus) uses the secretion of the chin gland to maintain dominance hierarchies in the wild. Recent work has investigated changes in the secretion when social status is manipulated in the rabbit. When a rabbit becomes dominant, a new compound appears in his secretion, 2-phenoxyethanol. This compound is used as a fixative in the perfume industry. This study investigates whether the compound performs a similar function in the secretion of the rabbit. 2-Phenoxyethanol is not detected olfactorially by rabbits, and slows the release rate of some of the compounds that occur naturally in rabbit chin gland secretion. We suggest that when a rabbit becomes dominant, he adds a fixative to his secretion so that his scent will persist in the environment and not dissipate. He will thus come to dominate the olfactory environment, in much the same way as he does the physical environment.     

Semiochemicals and social signaling in the wild European rabbit in Australia: II. Variations in chemical composition of chin gland secretion across sampling sites

The volatile components of the chin gland secretion of the wild European rabbit, Oryctolagus cuniculus (L.), were investigated with the use of gas chromatography. Studies of the chemical nature of this secretion by previous workers demonstrated that it was important in the maintenance of social structure in this species. This study identified 34 different volatile components that consist primarily of aromatic and aliphatic hydrocarbons. Especially common are a series of alkyl-substituted benzene derivatives that provide most of the compound diversity in the secretion. Samples of chin gland secretion collected from animals at three different geographical locations, separated by more than 100 km, showed significant differences in composition. This work suggests that variation among populations needs to be considered when undertaking semiochemical research. Alternate nonparametric methods are also used for the analysis of chromatographic data.     

A comparison of the effectiveness of the odors of rabbits,Oryctolagus cuniculus,in enhancing territorial confidence

In 14 series, each of 100 tests, involving 350 individual animals it has been shown that odors derived from the anal, inguinal and chin glands and urine strengthen to different degrees the confidence of European wild rabbits,Oryctolagus cuniculus. In the tests, two rabbits were put together on otherwise neutral ground previously treated with the odor from one of the contestants. The confidence of the rabbits was determined from an assessment of the record of the time to first physical contact, the frequencies of approaches and aggression, the number of attempts to escape, and also by making a subjective assessment of their general deportment. The results confirm earlier conclusions on the territorial functions of the anal and chin glands, the role of inguinal glands in individual identification, and the behavioral role of urine. On otherwise neutral ground the rabbits' own chin secretions were found to be the most effective in stimulating the confidence of males and that of their female partners. The odor of the anal gland secretions of males stimulated a similar but slightly weaker effect. The anal gland secretions of females influenced their own confidence but not that of males. Urine seemed to play a more important role in affecting the confidence of females than of males. Inguinal gland secretions had least effect. Odor from this source generally did not influence the confidence of the experimental animals; secretion from the inguinal glands of males had a slight effect on the behavior of their female partners.     

Alert odor from skin gland in deer

Black-tailed deer (Odocoileus hemionus columbianus) discharge an odor from the metatarsal (MT) gland, located on the hindleg, when disturbed or alarmed. Freely moving, captive deer were exposed to the MT odor by means of a remote-release apparatus. Responses by males and females to MT odor from both sexes were recorded with a coding system and a video camera. When the odor was present, females became more alert and left the site more often than in the presence of control odors, odorless air, or deer urine. It is concluded that the MT secretion provides an alert signal, placing the odor into the class of alarm pheromones. There is no evidence that the closely related white-tailed deer,O. virginianus, possesses this alert (or alarm) odor system to the same degree.     

Semiochemicals and Social Signaling in the Wild European Rabbit in Australia: I. Scent Profiles of Chin Gland Secretion from the Field

The European rabbit (Oryctolagus cuniculus) uses the secretion of the chin gland in the maintenance of social status. Previous work has concentrated on secretion collected directly from the animal. In this study, the analysis was conducted by collecting scent marks made by free-ranging animals. Scent marks were found to be concentrated at the center of the area controlled by a social group, and at the boundaries between two adjacent social groups. Only the mark from dominant animals could be identified. Marks were also collected from the skin of rabbits, where they had been placed by the dominant individual. The mark found on the head of a subordinate animal may, in the future, be used to identify the dominant animal of the social group, who placed the mark.     

Behavioral and cardiac responses of the rabbit,Oryctolagus cuniculus,to chemical fractions from anal gland

Unrestrained rabbits, 2–3 months old and nestlings, were exposed to the odors of the acid-neutral, acid, neutral, and basic fractions of the anal gland of male rabbits. The acid-neutral, acid, and neutral fractions elicited the strongest response in the form of avoidance. The heart rates of the nestlings were monitored using a radio telemetry technique. Statistically significant lowering of the heart rate and an increase in variability occurred on exposure to the fractions. The extent of the changes varied consistently in relation to a given odor, being greatest for the acid and neutral fractions. The results indicate the usefulness of heart-rate monitoring as a tool in the study of the perception of odors by animals.     

Influence of odor from wood-decaying fungi on host selection behavior of deathwatch beetle,Xestobium rufovillosum

Adult females of Xestobium rufovillosum de Geer demonstrated anemotactic orientation when exposed to an odor plume containing volatiles generated by wood-decaying fungi (Coriolus versicolor, Donkioporia expansa) and decayed oak wood (Quercus petraea, Quercus robur). They did not orient towards undecayed oak wood, beech (Fagus sylvatica), or pine wood (Pinus sylvestris). Although all insects tested showed anemotactic orientation, responses were nonlinear with respect to insect age. Adult females more readily oriented upwind when they were between 10 and 16 days old. Oviposition choice bioassays showed that ovipositing females would preferentially oviposit on extract-treated cellulose paper discs that had been treated with various strains of the wood-decaying fungus, Donkioporia expansa. HPLC-fractionated mycelial extracts were attractive to ovipositing deathwatch beetles, whereas HPLC-fractionated fungal broth extracts were repellent to ovipositing females. The implications of these results are discussed in the context of timber pest management in historically important buildings.     

The effect of short-range host odor stimuli on host fruit finding and feeding behavior of plum curculio adults (Coleoptera: Curculionidae)

In laboratory assays, we investigated responses of female plum curculios (PCs),Conotrachelus nenuphar (Herbst), to host and nonhost fruit or leaf odor when PCs were crawling on experimental tree branchlets or twigs. In choice tests where test specimens were hung from the ends of a wooden crosspiece, PCs made significantly more visits to host plum fruit than to plum leaves, nonhost tomato fruit, wax models of plum fruit, or blanks (wire). In similar tests, PCs made significantly more visits to plum leaves compared to nonhost maple leaves or to blanks. PCs in test chambers that contained host or nonhost odor were significantly more prone to feed on wax plum models in the presence of odor from host fruit or host leaves compared to odor from nonhost fruit or leaves or a water blank. In choice tests offering alternating cluster types on an apple branchlet, PCs visited leaf clusters bearing a host apple fruit more than leaf clusters without a fruit. In tests to assay the distance at which PCs can detect an individual host fruit, PCs crawled from the central stem of an apple branchlet onto a side stem significantly more often when an apple fruit on a side stem was hung 2 cm from the central stem compared to 4 or 8 cm away. Our combined results suggest that PCs use host fruit odor to locate host fruit at close range.     

Volatiles from feces of wild male house mice

Volatile compounds from the fecal pellets deposited by a house mouse could be used for the purpose of communication with conspecifics. Analysis of volatiles from fresh and aged pellets by gas chromatography-mass spectrometry showed that many compounds were present and that ketones, alcohols, and carboxylic acids were the most common. Introducing volatiles from the fecal pellets of strange males into the territory of a singly housed male altered the site at which the resident mouse deposited most of its feces, and introducing the fecal pellets of a male mouse into the environment where it encountered a conspecific improved its success in an encounter with a stranger. The combined volatiles from feces of strange males had a greater effect on the heart rate of a male mouse than the combined volatiles from its own feces. Several individual volatiles from the fecal pellets of strange males affected the heart rates of test animals, but individual animals responded differently to constituents with similar retention times.     

Bioassay approaches to assessing behavioral responses of plum curculio adults (Coleoptera: Curculionidae) to host fruit odor

We evaluated several approaches to developing a simple, sensitive, and reliable laboratory bioassay of responses of overwintered adult plum curculios (PCs),Conotrachelus nenuphar (Herbst), to host fruit odor or its attractive components. A high proportion of assayed PCs responded positively to odor of wild plums under no-choice, moving-air conditions in a wind tunnel and under dual-choice, still-air conditions in enclosed Petri dishes. Positive response to controls lacking host odor, however, was much greater in the wind tunnel, arguing in favor of bioassays under dual-choice conditions in still air to provide greater PC discrimination. Response to host odor (from wild plums or hexane extract of wild plums or Liberty apples) in Petri dish bioassay chambers proved greatest: (1) during the scotophase of PCs under total dark or dim red light conditions, (2) when Petri dishes were completely enclosed, (3) when PCs were starved for 24 or 48 hr, and (4) when PCs were tested within seven weeks after apple tree petal fall. Neither the sex of a PC nor the direction in which a PC was obliged to move to find the source of host odor (upward through a port in the Petri dish lid or downward through a port in the base) had a substantial effect on level of response to host odor or discrimination of host odor from a nonodorous control. We conclude that an enclosed Petri dish bioassay chamber of the type described here should be a valuable asset in the process of chemically identifying components of host fruit odor attractive to PCs.     

A comparison of the effectiveness of predator odor and plant antifeedant in deterring small mammal feeding damage on lodgepole pine seedlings

The influence of mustelid (weasel) anal-gland compounds (semiochemicals) and a plant antifeedant (pinosylvin) on snowshoe hare (Lepus americanus) and meadow vole (Microtus pennsylvanicus) feeding damage was investigated. This study has shown conclusively that when applied to spring-planted lodgepole pine seedlings, pinosylvin is effective in significantly reducing feeding damage. Seedlings treated with pinosylvin suffered damage levels of 2.3% and 21.2% compared to the controls with damage levels of 24.5% and 41.0%. Weasel scent was found to reduce damage levels, but the reduction was not statistically significant in all blocks.     

Attraction ofCeratitis capitata (Diptera: Tephritidae) flies to odor of coffee fruit

On potted nonfruiting host trees in outdoor field cages, we evaluated attraction of released mature laboratory-cultured or wild-origin Mediterranean fruit flies (medflies) to odor of freshly picked fruit of host and nonhost plants. Odor of ripe intact or crushed coffee fruit (the presumed ancestral host of medflies) was significantly more attractive than odor of ripe intact or crushed fruit of five lower-ranking hosts and three nonhosts. Odor of crushed coffee fruit was significantly more attractive than odor of intact coffee fruit. Odor of ripe or near-ripe coffee fruit was significantly more attractive than odor of unripe coffee fruit. Immature females (without eggs) were significantly more attracted to odor of a proteinaceous food lure than to odor of ripe coffee fruit, whereas the reverse was true for mature females carrying a high egg load. In some trials, males proved as discriminating as females in favor of coffee fruit odor, but in several other trials males were less discriminating than females. Response patterns of mature laboratory-cultured females were similar to those of mature wild-origin females. In a field of coffee plants, attraction of natural-population females was significantly greater to odor of ripe coffee fruit than to water but was not greater than attraction to odor of proteinaceous food. Findings are discussed in relation to potential use of synthetic volatiles of coffee or other host fruit in traps for monitoring or controlling medflies.