Analysis of secretions from scent-producing glands of brushtail possum (Trichosorus vulpecula Kerr)

A characterization at a molecular level of the chemical composition of the secretions of the two pairs of paracloacal glands and of the sternal sebum of the brushtail possum (Trichosorus vulpecula Kerr) has been undertaken with a view to evaluating the potential of volatile pheromone components as species-specific attractants for use in novel baiting systems. Particular attention has been given to the respective fatty acid fractions produced by chemical hydrolysis, since these are believed to be the products of postemission microbial degradation (fermentation) of the secretions. In all instances, the highly complex distribution of the constituents present in these organic components of the secretions were shown to be virtually identical in adult males and females. A unique suite of low-molecular-weight branched-chain carboxylic acids has been shown to be produced by chemical degradation of the holocrine (oil-secreting) gland secretion. This odor signature is suggested to function as a unique scenting-the-habitat pheromone that might act as an attractant to all members of the species.     

Cover and efficacy of predator-based repellents for Townsend's voleMicrotus townsendii

Predator-based repellents have been used experimentally to control wildlife damage in both agriculture and forestry, but they have not always been effective. We examined the relative importance of cover and predator odors in forage patch selection by Townsend's vole,Microtus townsendii, and its behavior related to cover and predator cues. Experiments were conducted in which forage patch and area choices were related to available habitat alternatives. Outdoor enclosures were divided into halves: one side was treated and the other used as a control. Treatments consisting of cover, repellent, and cover plus repellent were compared to controls (no cover, no repellent). In the absence of cover, voles preferred to feed on the side without repellents. When cover was present, voles preferred to feed on the side with cover, regardless of whether or not repellents were present. Voles visited more feeding stations on the side without cover when repellents and cover were present than they visited during cover-only treatments. These additional feeding stations, visited outside of cover, were used only lightly as food sources. The amounts of oats eaten by voles decreased with increasing distance from cover. This inverse relationship had a steeper slope in coveronly treatments compared to cover plus repellent treatments. A selection model based on forage patch selection and a habitat preference hierarchy is proposed. We conclude that predator odors are effective as repellents, but their efficacy depends on habitat conditions. Managers intending to use predator-based repellents must ensure that alternative sites available to pests are better quality habitat than in areas to be protected.     

Avoidance of bird repellents by mice (Mus musculus)

It is believed that mammalian chemosensory irritants are not aversive to birds and vice versa. Nevertheless, few avian repellents have been tested against mammals. For that reason, we evaluated the efficacy of 1.0% w/v methyl anthranilate, orthoaminoacetophenone, 2-amino-4,5-methoxyacetophenone, 2-methoxyacetophenone, and veratryl amine as mouse repellents in 3-hr no-choice drinking tests. Relative to ingestion of plain water, all test substances significantly reduced (P < 0.05) intake. Orthoaminoaceto-phenone was the most effective repellent, with intake reduced to levels statistically indistinguishable from zero.     

Oviposition attractants and repellents of mosquitoes

Infusions of various organic materials have been known to modify ovipositional behavior of gravid female mosquitoes. Previously, we found that an infusion of Purina Laboratory Chow elicited negative ovipositional behavior inCulex pipiens quinquefasciatus Say andCulex tarsalis Coquillett. In the present chemical studies, the Purina Laboratory Chow infusion, fermented for 10 days, was distilled to give an active distillate which, upon ether extraction, gave an active ether extract. Fractionation of the ether extract yielded an active acidic fraction and inactive nonacidic fraction. Gas Chromatographie analysis on Porapak R and AT-1200-H₃PO₄ columns of the acidic fraction showed the presence of acetic, propionic, isobutyric, butyric, isovaleric, and caproic acids. In bioassay tests, these lower aliphatic carboxylic acids, individually and in combination, exhibited ovipositional repellency against the two species of mosquitoes at the concentration of 6 × 10^–2%. At this concentration, butyric acid caused 100% mortality in first-instar larvae ofC. p. quinquefasciatus. Gravid female mosquitoes might have acquired, through evolutional adaptation, the ability of avoiding ovipositing in unsuitable sites in which toxic compounds might be detrimental to the survival and development of their offspring.Diptera: Culicidae.     

Carnivore fecal chemicals suppress feeding by Alpine goats (Capra hircus)

The efficacy of carnivore and ungulate fecal chemicals in suppressing the feeding behavior of Alpine goats (Capra hircus) was examined. In the first four experiments, goats were offered food covered with paper strips treated with fecal extracts of the Bengal tiger, Siberian tiger, African lion, and brown bear, respectively; food covered with solvent-treated and untreated (plain) papers served as controls in each experiment. Goats made fewer head entries into, and ate less food from, buckets containing fecal extracts. In the fifth experiment, goats were offered food covered with paper strips treated with fecal extracts of the puma, Dorcas gazelle, white-bearded gnu, and conspecifics; food covered with solvent-treated and plain papers again served as controls. The amounts of food consumed from buckets containing puma, gazelle, gnu, and solvent treatments were statistically indistinguishable, but less food was consumed from them than from buckets containing the goat-scented or plain papers. No significant differences among treatments were detected with respect to head entries. Field experiments are needed on the use of predator-derived chemicals to reduce damage by goats to vegetation.     

Oviposition repellents of mosquitoes: Negative responses elicited by lower aliphatic carboxylic acids

Organic infusions have been shown to elicit discriminatory responses in ovipositing mosquitoes. Previously, we found that a Purina® Lab Chow infusion induces negative oviposition behavior inCulex pipiens quinquefasciatus Say andCulex tarsalis Coquillett. Six aliphatic carboxylic acids isolated from the active fraction of this infusion were acetic, propionic, isobutyric, butyric, isovaleric, and caproic. In the present studies, we have quantified the negative oviposition responses ofCulex mosquitoes to these carboxylic acids in a laboratory bioassay system and have also tested the main acid component, butyric acid, againstCuliseta, Aedes, andAnopheles mosquitoes.     

Use of predator odors as repellents to reduce feeding damage by herbivores

The effectiveness of predator odors (fecal, urine, and anal scent gland) in suppressing feeding damage by snowshoe hares was investigated in pen bioassays at the University of British Columbia Research Forest, Maple Ridge, British Columbia, Canada. A total of 28 bioassay trials tested the effects of these odors on hare consumption of willow browse and coniferous seedlings. Lynx and bobcat feces, weasel anal gland secretion, and lynx, bobcat, wolf, coyote, fox, and wolverine urines resulted in the most effective suppression of hare feeding damage. Novel odors of domestic dog urine and 2-methylbutyric acid did not reduce feeding. A field bioassay with lodgepole pine seedlings and weasel scent provided significant results comparable to the pen bioassays. The short-term (up to seven days) effectiveness of these treatments was more likely due to evaporative loss of the active repellent components of a given odor than habituation of hares to the stimulus. Predator odors as repellents have a biological basis compared with the anthropomorphic origins of commercial repellents. When encapsulated in weather-proof controlled-release devices, these odors could provide long-term protection for forestry plantations and agricultural crops which experience hare/rabbit feeding damage.     

Constraint of Feeding by Chronic Ingestion of 1,8-Cineole in the Brushtail Possum (Trichosurus vulpecula)

Eucalyptus leaf-eating marsupials such as the brushtail possum (Trichosurus vulpecula) ingest large amounts of terpenes, especially 1,8-cineole (cineole)--the major component of many eucalyptus oils. Brushtail possums were acclimated to a non-Eucalyptus diet with increasing concentrations of cineole (0.5-4.0% wet weight) added over 18 d. We measured food and cineole consumption and urinary metabolites of cineole. Food intake decreased with cineole content, indicating that it was constrained by the maximum tolerable intake of cineole that was 3.8 +/- 0.2 g kg(-1) or 5.2 +/- 0.3 g kg(-0.75) (mean +/- SE, N = 6). The pattern of metabolites was similar at all cineole intakes (56% hydroxycineolic acids, 27% cineolic acids, 13% hydroxycineoles, and 4% dihydroxycineoles). In another experiment, possums maintained on artificial diet were abruptly presented with 4% cineole for 5 d. Food intake fell by 45 +/- 6% (mean +/- SE, N = 6) and mean cineole intake was 2.9 +/- 0.3 g kg(-1). There was evidence of induction of secondary oxidative pathways, as hydroxycineoles were the major metabolites (48% total) on the first day, but rapidly dropped to 15% on subsequent days as the acid metabolites increased. These findings indicate that ingestion of cineole is not constrained by selective saturation of individual enzymes involved in its multiple pathways of oxidation, but rather the total detoxification capacity appears to limit feeding on a cineole diet.     

Use of predator odors as repellents to reduce feeding damage by herbivores

This study investigated the influence of the major anal-gland compounds from the stoat (Mustela erminea) and fecal and urine compounds from the red fox (Vulpes vulpes) in generating an avoidance response by montane voles (Microtus montanus), as well as suppressing feeding by montane and meadow (M. pennsylvanicus) voles on apple trees in orchards. In trap bioassays, a 11 mixture of 2-propylthietane and 3-propyl-1,2-dithiolane significantly reduced vole captures. Other mixtures of stoat compounds reduced the number of new voles captured but not total individuals. 2,5-Dihydro-2,4,5-trimethylthiazoline, a component of fox feces, significantly reduced vole captures in one of two bioassays. Deer mice (Peromyscus maniculatus) did not show a negative response to any predator odor. In overwinter field bioassays, mixtures of 2-propylthietane and 3-propyl-1,2-dithiolane clearly reduced vole feeding on apple trees in four test blocks. 2,5-Dihydro-2,4,5-trimethylthiazoline and a synthetic fox urine mixture also significantly reduced vole attack in respective orchard blocks. Similarly, the intensity of vole feeding, in terms of amount of bark and vascular tissues removed from trees, was reduced by 60% to 97% in predator odor treatments compared with the control. Our study reports the first long-term (four to five months) use of synthetic semiochemicals as area repellents for crop protection from vole feeding damage.     

Tests and refinements of a general structure-activity model for avian repellents

We tested the robustness of a structure-activity model for avian trigeminal chemoirritants. Fourteen benzoates and acetophenones were tested using European starlingsSturnus vulgaris as a bioassay. In general, the previously proposed model was a reasonable predictor of repellency (i.e., irritant potency). We found that the presence of a phenyl ring was critical to repellency. Basicity of the molecule is the next most critical feature influencing repellency. The presence of an acidic function within the electron-withdrawing functionality seriously detracts from repellency. The presence or absence of an electron-withdrawing or -donating group may potentiate repellent effects, but its presence is not critical, so long as the phenyl ring is electron rich. Our data suggest that there is ano-aminoacetophenone/methyl anthranilate trigeminal chemoreceptor in birds analogous to the mammalian capsaicin receptor. Both receptors contain a benzene site. However, birds seem to lack the associated thiol/hydrogen-bonding site present in mammals which is needed to activate the benzene site. Rather, birds may possess an associated exposed charged site that in turn may interact with the stimulus to activate the benzene site. These differences may explain the differential sensitivity of birds and mammals to aromatic irritants.     

Structure-activity relationship of unsaturated fatty acids as mosquito ovipositional repellents

Various straight-chain unsaturated fatty acids from C₁₄ to C₂₄ were evaluated for their ovipositional repellency against gravid females of the southern house mosquitoCulex quinquefasciatus Say, and the relationship between the structures of the fatty acids and their ovipositional repellency was determined. A double bond withZ configuration was prerequisite for an unsaturated fatty acid to be highly repellent;E isomers were less active or even inactive. No relationship was found between the repellency and the number of double bonds in the unsaturated fatty acids. In C₁₈ monounsaturated fatty acids, (Z)-9 acid was more active than (Z)-11 and (Z)-6 acids, indicating that a double bond at the 9 position rendered an acid highly repellent. Among (Z)-9-alkenoic acids of different chain lengths, the most repellent was C₁₈ acid which was also more active than (Z)-11-C₂₀, (Z)-13-C₂₂, and (Z)-15-C₂₄ acids. Oleic[(Z)-9-octadecenoic]acid, which met all these criteria, was the most ovipositionally repellent among the unsaturated fatty acids tested.Diptera: Culicidae.     

Germacrone defends labrador tea from browsing by snowshoe hares

Labrador tea (Ledum groenlandicum), a slow-growing late successional evergreen, is highly unpalatable to snowshoe hares (Lepus americanus). Germacrone, a sesquiterpene that is the major component of the essential oil ofL. groenlandicum, was shown by bioassay to be a potent antifeedant to hares. Its concentrations in leaves and intemodes of the plant are high enough to defendL. groenlandicum from hares. This chemical defense of Labrador tea from herbivory is consistent with the resource availability theory of antiherbivore defense.     

Responses of Mud Snails and Periwinkles to Environmental Odors and Disaccharide Mimics of Fish Odor

Estuarine snails, periwinkles (Littoraria irorata), and mud snails (Ilyanassa obsoleta) were tested for behavioral responses to aqueous extracts of tissue macerates, odors of living intact organisms, and to disaccharides derived from heparin. Extracts included salt-marsh cordgrass (Spartina alterniflora), pink shrimp (Penaeus duorarum), crushed periwinkles, and crushed mud snails. Odors included live periwinkles, mud snails, stone crab (Menippe mercenaria), striped hermit crab (Clibanarius vittatus), tulip snail (Fasciolaria hunteria), and mummichog (Fundulus heteroclitus). Responses were determined upon contact by snails of a ring of solution in the bottom of an otherwise dry bowl and by presenting snails in seawater with 25 l of solution on a cotton swab. In each test, the response of 30 individuals was determined. Behaviors were recorded as fed, alarm, and no response. The strongest alarm response (>80% of all snails tested) in both snails was elicited by crushed mud snails. The strongest feeding response was to shrimp and periwinkle extract for mud snails (>70%), and salt-marsh cordgrass extract attracted periwinkles. High percentages of mud snails and periwinkles fled in response to the odors of intact snail predators, stone crabs, tulip snails, and mummichogs. Mud snails and periwinkles did not flee in response to nonpredator odors, including each other's odor, as well as that of hermit crabs, shrimp, and marsh cordgrass. Heparin disaccharides were tested because other studies reported that biological activity of predatory fish odor is due to similar disaccharides originating from fish mucus. Mud snail responses to disaccharides indicate that they respond to the same kinds of molecules as crustacean larvae and that the modified amine on the disaccharide is essential for activity. The Q-tip assay is appropriate for bioassay-directed purification of alarm signals originating in fish body odor.     

Multivariate study of moose browsing in relation to phenol pattern in pine needles

Extracts of needles from Scots pine,Pinus sylvestris L., grown on fertile (A) or poor (B) soil, and from lodgepole pine,P. contorta Doug. (C), have been analyzed with HPLC and UV detection. By application of multivariate analysis, the phenol pattern was shown to vary between species and between habitats. Within groups A and B, moose food choice was highly correlated to phenol pattern. Concentrations of most of the analyzed compounds were internally correlated. Information related to pine growth rate could also be extracted from the phenolic pattern for A, B, and C. Moose food choice was not correlated to pine growth rate.     

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.     

Selected chemical constituents and deer browsing preference of Douglas fir

Douglas fir foliage from eight full-sib F₁ families was analyzed for chlorogenic acid and essential oils and ranked according to browsing preference of penned black-tailed deer. Foliage was obtained during the dormant season, and tissues for both the preference test and chemical analyses were collected from the same trees. Deer ranked the different families, and chlorogenic acid content was correlated with preference order. Families varied significantly in yield and composition of essential oils, but differences were not related to preference. Results may have application in selections for Douglas fir resistant to browsing.     

Odors as cues for orientation to mothers by weanling Virginia opossums

Three experiments were conducted to investigate whether wholebody and pouch odors facilitate social cohesion between young Virginia opossums and their mothers just prior to weaning. In experiment 1, young oriented toward plastic buckets containing their mothers, directing significantly higher levels of investigative behavior and more distress vocalizations toward them than toward buckets containing unrelated lactating females. In experiment 2, young oriented toward and investigated empty buckets containing whole-body odors of their mothers more than empty buckets containing odors of other females. Similarly, more investigative behavior was directed toward plastic bucket lids containing pouch odors from subjects' mothers than toward pouch odors from unrelated females in experiment 3. These results suggest that social odors help young didelphid marsupials maintain contact with their mothers, as in other mammals, and that whole-body and pouch gland odors are important chemical signals in this nongregarious species.     

Effects of birch phenolics on a grazing and a browsing mammal: A comparison of hares

Mountain hares (Lepus timidus L.) commonly have high proportions of birch (Betula spp.) in their winter diets, whereas European hares (Lepus europaeus) do not. The effects of a birch extract added to laboratory diets offered to mountain hares and European hares on the digestibility and sodium balance were measured. The extract added contained total phenolics equivalent in amount to that occurring in diets containing 0, 40, 60 and 80% dry matter of birch twigs. Increasing dietary phenolic concentration led to reduced voluntary food intake and apparent protein digestibility in both hare species. No effects on apparent dry matter digestibility were observed. The highest concentration of birch extract caused severe sodium losses via the urine by European hares but not by mountain hares. The effects of the birch extract within the digestive system appear to be similar in the two hare species, whereas the mountain hare appears to be better adapted to the toxic effects that disrupted sodium balance in the European hare. These preliminary results suggest that detoxification rather than digestive abilityper se may contribute to different mammalian herbivores' feeding strategies.     

The processing of new oilseed crops—An economic evaluation

The economics of pilot- to large-scale processing of the new oilssed cropsCrambe abyssinica, Euphorbia lagascae, andDimorphotheca pluvialis into oil, fatty acids, or esters were estimated. It was found that the processing costs forCrambe seed to oil is in the same range as that for rapeseed (≈0.5 U.S. $/kg). Production of fatty acid esters from vernolic and dimorphecolic acids requires some special downstream processing operations that result in processing costs of about 1 U.S. $/kg ester. Good-qualityDimorphotheca oil is much more difficult to obtain and requires supercritical carbon dioxide extraction. Processing costs can be as high as 4 U.S. $/kg oil.     

Inheritance Of Resistance to Mammalian Herbivores and of Plant Defensive Chemistry in an <Emphasis Type="Italic">Eucalyptus</Emphasis> Species

Hybridization in plants provides an opportunity to investigate the patterns of inheritance of hybrid resistance to herbivores, and of the plant mechanisms conferring this resistance such as plant secondary metabolites. We investigated how inter-race differences in resistance of Eucalyptus globulus to a generalist mammalian herbivore, Trichosurus vulpecula, are inherited in their F₁ hybrids. We assessed browsing damage of 3-year-old trees in a common environment field trial on four hybrid types of known progeny. The progeny were artificial intra-race crosses and reciprocal inter-race F₁ hybrids of two geographically distinct populations (races) of E. globulus north-eastern Tasmania and south-eastern Tasmania. Populations of trees from north-eastern Tasmania are relatively susceptible to browsing by T. vulpecula, while populations from south-eastern Tasmania are more resistant. We assessed the preferences of these trees in a series of paired feeding trials with captive animals to test the field trial results and also investigated the patterns of inheritance of plant secondary metabolites. Our results demonstrated that the phenotypic expression of resistance of the inter-race F₁ hybrids supported the additive pattern of inheritance, as these hybrids were intermediate in resistance compared to the pure parental hybrids. The expression of plant secondary metabolites in the F₁ hybrids varied among major groups of individual compounds. The most common pattern supported was dominance towards one of the parental types. Together, condensed tannins and essential oils appeared to explain the observed patterns of resistance among the four hybrid types. While both chemical groups were inherited in a dominant manner in the inter-race F₁ hybrids, the direction of dominance was opposite. Their combined concentration, however, was inherited in an additive manner, consistent with the phenotypic differences in browsing.