Tongue-flicking and biting in response to chemical food stimuli by an iguanid lizard (Dipsosaurus dorsalis) having sealed vomeronasal ducts: Vomerolfaction may mediate these behavioral responses

In the iguanid lizardDipsosaurus dorsalis, chemical food stimuli were discriminated from other odorants by vomerolfaction. This was demonstrated in a 2 × 3 experiment in which groups of lizards with sealed vomeronasal ducts or sham-sealed vomeronasal ducts responded to carrot chemical stimuli, cologne, and distilled water presented on cotton-tipped applicators. Abilities to detect and discriminate food chemicals were abolished in lizards having sealed vomeronasal ducts. For tongue-flick attack score and number of lizards biting, the sham-sealed group responded more strongly to carrot stimuli than to the control stimuli, but the group having sealed ducts did not. Lizards having sham-sealed ducts responded more strongly to carrot stimuli than did lizards having sealed ducts; responses by the two groups of lizards to control stimuli did not differ. Tongue-flicking occurred when the vomeronasal system detected a chemical stimulus from either carrot or cologne. Biting occurred only when the vomeronasal organ detected food stimuli (from carrot). Most duct-sealed lizards opened their mouths, some repeatedly. Mouth-opening thus occurs when the vomeronasal organ does not detect chemicals. It may be an attempt to stimulate or prime the vomeronasal organ or to dislodge the sealant.     

Orientation of the fiddler crab,Uca cumulanta: responses to chemical and visual cues

Behavioral responses of the fiddler crab Uca cumulanta to flat geometric shapes mimicking natural objects were measured in a circular arena by using zonal recovery as a behavioral measurement. Crabs were tested either in presence or absence of odors from two common predator species, the blue crab Callinectes sapidus, and the pufferfish Sphoeroides testudineus. The study tested the hypothesis that U. cumulanta have different behavioral responses to visual cues in the presence of chemical cues associated with predators. Escape direction tests demonstrated that U. cumulanta is able to show zonal recovery behavior based upon astronomical references. When tested in water lacking predator odor, crabs failed to exhibit a consistent orientation if a single silhouette target was interposed in the landward direction. However, when animals were tested in different predator odor concentrations, an orientation response was obtained at 10 and 20 g/liter/hr blue crab odor and 10 g/liter/hr pufferfish odor, demonstrating U. cumulanta ability to detect the potential presence of its natural predators by this odor. Thus, the hypothesis was supported, and the results suggest that behavioral responses to chemical and visual cues are involved in predator avoidance.     

Predatory response of brown tree snakes to chemical stimuli from human skin

The brown tree snake (Boiga irregularis) is an exotic pest species on Pacific islands, most notably on Guam where it has caused considerable ecological and economic damage. On Guam, the snake commonly associates with people and can be found near or in human habitations. Bites are common, approximately 1 of 1200 emergency room visits to Guam hospitals were reported to be the result of B. irregularis bites; 80% of these victims were attacked while sleeping. Most of the attacks occurred on fingers and hands and the attacks appeared to be predatory, rather than defensive, in nature. In order to characterize the mechanism releasing this unusual behavior, we measured the predatory response of B. irregularis to chemical stimuli from humans and controls using a lab population that originated from Guam and a wild population from the species' native range in Queensland, Australia. To quantify behavior we measured the proportion of snakes displaying predatory behavior to each of the stimuli, the latency to attack, and the number of tongue-flicks displayed. We quantified predatory behavior using the tongue-flick attack score for repeated measures [TFAS(R)], a common method for quantifying predatory behavior in squamate reptiles. Captive brown tree snakes responded to human skin stimuli with feeding behavior, including predatory attacks, at the same frequency as they did to prey stimuli derived from mice, while never responding to controls with such behavior. Captive snakes also responded to human skin stimuli and prey stimuli with significantly higher TFAS(R) scores than to controls, although there were no differences between the human and mouse stimuli. Wild-caught animals in Australia also responded with predatory attacks to human skin stimuli, while not showing predatory behavior to a blank control and with higher TFAS(R) scores to human skin stimuli than to the control. As B. irregularis is a generalized predator that relies heavily on chemical signals to recognize prey, we hypothesize that the snakes recognize compounds on human skin that may be shared with other prey.     

Effects of predator chemical cues and behavioral biorhythms on foraging activity of terrestrial salamanders

Red-backed salamanders, Plethodon cinereus, show a variety of alarm responses to chemical cues from eastern garter snakes, Thamnophis sirtalis. We measured the foraging activity of red-backed salamanders exposed to water soiled by a garter snake (fed P. cinereus) or to unsoiled water. Salamanders exposed to snake-soiled water showed less foraging activity than salamanders exposed to unsoiled water; therefore, predators could have nonlethal effects on salamander populations. Our results also show additional factors influenced salamander foraging activity. Salamander foraging activity and responsiveness to chemical cues do not appear to have been affected by sex or food deprivation. Salamander foraging activity does appear to have been influenced by activity biorhythms. Foraging activity of animals in both treatments showed a bimodal periodicity that is consistent with natural activity patterns controlled by internal biorhythms. Exposure to snake-soiled water significantly reduced foraging activity during periods of peak foraging activity, but had a subtler effect0 on foraging activity during natural lulls in activity. We suggest that both activity biorhythms and exposure to chemical cues are important factors affecting salamander foraging behavior.     

Timber Rattlesnakes (Crotalus horridus) Use Chemical Cues to Select Ambush Sites

Chemicals left by organisms moving through the environment are used by other organisms to mediate interspecific interactions. Most studies of chemical eavesdropping focus on prey responding to chemical cues from predators, despite the fact that chemical cues are frequently used by predators as a source of information about prey. Crotalus horridus uses a foraging strategy that is widespread among sedentary predators: the snake chooses a site where it is likely to encounter prey and remains immobile for many hours. I investigated this ambush hunting behavior in captive-raised timber rattlesnakes and provide evidence that sit-and-wait predators may discriminate among prey chemical cues, even when they have no prior experience with the prey. Snakes explored chemical cues with chemosensory behaviors, and more frequently adopted a stereotyped ambush foraging posture toward chemical cues from prey sympatric with their population of origin than either allopatric prey or sympatric nonprey species that are eaten by other viperids. These results support the notion that intra- and interspecific variation in diet may be mediated proximally by innate recognition of cues from particular prey items. This system also describes a bioassay that may be used in the isolation and identification of prey-derived kairomones. Studies such as this can be used to determine more realistic parameters for models of predator-prey interaction and foraging behavior that involve secretive, less active predators.     

Following of conspecific and avoidance of predator chemical cues by pine snakes (Pituophis melanoleucus)

The ability of hatchling pine snakes (Pituophis melanoleucus) to follow or avoid the chemical trails of conspecifics and a king snake (Lampropeltis getulus) on paper substrates was investigated inY-maze experiments. Hatchlings entered the arm with the adult conspecific trail and avoided the arm containing the king snake trail at a frequency much greater than that due to chance. The data support the hypotheses that pine snakes follow the chemical trails of adult conspecifics and avoid the chemical trails of a predator.     

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.     

Responses of ecologically dissimilar populations of the water snakeNatrix s. sipedon to chemical cues from prey

Three populations of the water snake,Natrix s. sipedon, were tested for their responses to surface water extracts prepared from various prey species. It was clearly shown that these snakes can distinguish between different prey genera on the basis of chemical cues alone. Adult snakes caught at a fish hatchery where goldfish (Carassius auratus) were abundant responded most to goldfish extract, as did laboratory-born snakes reared on goldfish for one year. However, snakes caught in a relatively undisturbed mountain stream environment and naive young born to a mountain female responded more to the extracts of various sympatric-mountain fish and amphibians. The evidence indicates that newborn snakes have genetic predispositions for sympatric species of the classes of prey normally eaten. These preferences can be enhanced or inhibited during ontogeny.     

Population and sex differences in antipredator responses of breeding fathead minnows (Pimephales promelas) to chemical stimuli from garter snakes (Thamnophis radix andT. sirtalis)

We conducted a predator bite survey on a population of fathead minnows (Pimephales promelas) considered to be under substantial predation pressure by western plains garter snakes (Thamnophis radix). Scarring, due to failed predation attempts by garter snakes and crayfish (Orconectes virilis), was observed significantly more often in breeding males than in breeding females and nonbreeding minnows. Likely, territorial nest defense under the edges of rocks along the water's edge, a habitat occupied by crayfish and frequented by snakes, caused the breeding males to be differentially vulnerable to predation. Under controlled laboratory conditions, breeding males from this population exhibited an antipredator response to chemical stimuli from live snakes (T. sirtalis andT. radix) significantly more often than breeding female minnows from the same population and breeding minnows of both sexes from a population that was presumed to be under lower predation pressure from snakes.     

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.     

Effect of birch (Betula pendula) bark and food protein level on root voles (Microtus oeconomus): I. food consumption,growth, and mortality

The effect of protein and birch bark powder (BBP) content of forage on food consumption and growth of root voles (Microtus oeconomus) was studied by feeding young voles with eight different diets for two weeks. Voles ate more when the protein content of forage was moderate (6%) or low (3%), and when BBP was added to the diets, as compared to a high (12%) protein diet without BBP addition. On high and moderate protein diets voles gained weight, whereas on low protein diets they lost weight. Addition of BBP caused reduced growth or weight loss. Consuming a diet with moderate (6%) protein and high BBP (20%) resulted in high mortality (9 of 10 died). These results show that root voles need at least 6% protein for growth and that the effect of birch bark depends on the protein content of the forage.     

Disturbance and reattachment behavior of sea anemonesCalliactis tricolor (Le Sueur): Temporal,textural and chemical mediation

Two hermit crabs,Dardanus venosus (Milne-Edwards) andPagurus pollicaris Say, have evolved mutualistic symbiotic relationships with sea anemones,Calliactis tricolor (Le Sueur). Anemones live on gastropod shells occupied by the crabs. Both partners are thought to participate in the transfer of the anemone to the crab's shell. However, neither species invariably exhibits transfer behavior when in contact with the other symbiont. Factors influencing transfer activity ofC. tricolor were studied. Anemone attachment is stimulated by disturbance and is time dependent. Anemones attached to shells more frequently immediately after the anemone was disturbed. Assays testing surface texture and surface composition showed that attachment activity of the sea anemone is dependent upon the organic matrix of the shell and is enhanced by texture. Attachment by anemones to shells with organic matrix removed is reduced, but is stimulated by adsorption of complex odor mixtures. Anemones respond to odor mixtures that signal food to crabs and to odor mixtures that stimulate shell investigation and shell switching in crabs. Thus,C. tricolor participates in shell transfer activity after disturbance. Behavior of the anemone is cued by texture and chemicals, including chemical mixtures that cause hermit crabs to change shells.     

Sheep food repellents: Efficacy of various products,habituation, and social facilitation

The study of food repulsion in ungulates is of practical importance for controlling browsing damage to forest trees and agricultural crops. Experiments with domestic sheep assessed the repulsive value of three natural and two synthetic products, using a food-choice situation. Animals were individually presented with two troughs, each containing 30 g of maize, one treated with a chemical product, and the other a control. The odor of domestic dog feces appeared to be highly repulsive, as none of the treated food was eaten, whereas 3 g ±9 and 6 g ±10 were eaten, respectively, of food treated with fetal fluids from sheep and odor of pig feces. Synthetic odors of lion feces and a commercial deer repellent were less efficient as repellents with 11 g ±13 and 14 g ±15 of food eaten, respectively. Habituation to three of the repellents was tested for a minimum of seven successive days. With dog feces, no habituation to the odor was observed. On the contrary, significant habituation (P < 0.05) was observed as early as the third trial for sheep fetal fluids and the fourth trial for the commercial repellent. To test for the effects of social facilitation, groups of four sheep including no, one, or two anosmic animals were presented with food treated with the odor of dog feces. No social facilitation was observed as none of the intact sheep ate any of the food, although the anosmic animals actively ate it throughout the test. In individual food choice tests, the odor of dog feces appeared to be an especially efficient repellent when compared with the other natural or synthetic products. Such repulsion was nearly complete, and neither habituation nor social facilitation could be observed.     

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

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

Behavioral responses of crayfish (Orconectes virilis andOrconectes rusticus) to chemical feeding stimulants

We conducted two experiments to assess how chemical stimuli affect feeding behavior, grooming, and walking in the crayfishesOrconectes virilis andOrconectes rusticus. In the first experiment,O. virilis was tested with 29 amino acids; in the second experiment,0. rusticus was tested with 12 amino acids, 13 additional single compounds, and two six-compound mixtures. InO. virilis, the following amino acids, in order of potency, elicited feeding movements:l-isoleucine, glycine, hydroxy-l-proline,l-glutamate,l-valine, and B-alanine. Grooming increased in response tol-phenylalanine,l-tryptophan,l-tyrosine,l-leucine,l-methionine, and D-aspartate. InO. rusticus, both mixtures and the following single compounds, in order of potency, elicited feeding movements: cellobiose, sucrose, glycine, maltose, glycogen, nicotinic acid methyl ester, putrescine, andl-glutamate. Grooming increased in response to putrescine only, and walking increased in response to glycogen only. The responsiveness of these crayfishes to a wide variety of chemicals may reflect the omnivorous foraging habits of these crustaceans.     

Chemical mediation of behavior in hermit crabs: Alarm and aggregation cues

Field studies with three common local species of hermit crabs,Clibanarius vittatus, Pagurus longicarpus, andPagurus pollicaris, showed that these crabs responded behaviorally to chemicals originating from crushed conspecifics. Hermit crabs are attracted specifically and in a manner similar to previously reported crab responses to odors from dead gastropods. Responses byC. vittatus to both kinds of odor are of three types: (1) aggregation/shell investigation responses (previously reported for odors from dead gastropods), characterized by increased locomotor activity, investigation of shells in the vicinity, and switching into empty shells; (2) alarm responses, in which crabs flee the area; and (3) withdrawal responses, in which crabs pull into their shells and do not come out. Studies withC. vittatus showed that the stimulatory chemicals originate from hemolymph, are less than 500 D, adsorb to octadecyl silica, and are recovered by elution with 20% methanol. Responses ofC. vittatus are dependent upon crab size, type of shell occupied, and shell fit. Chemicals originating from dead conspecifics provide a forum for shell acquisition by crabs in relatively small shells and alarm by crabs in relatively large shells.     

Chemical fractionation of shrimp extracts inducing bottom food search behavior in cod (Gadus morhua L.)

The bottom food search (BFS) feeding behavior in cod (Gadus morhua L.), has been used in a bioassay for chemical isolation of the feeding stimulant substances present in shrimp (Pandalus borealis). An aqueous methanol extract of ground shrimp was separated into acidic, neutral, and amphoteric/basic fractions by ion-exchange chromatography and into single components by preparative high-pressure liquid chromatography (HPLC). Of the isolated single components, the amino acid glycine was most potent, followed by alanine. Two unidentified substances were also highly potent. There was a synergistic effect between glycine, alanine, proline, and arginine. These four amino acids were more potent than the total amino acid pool found in the shrimp extract, indicating that there may be amino acids in this pool having an antagonistic effect.     

Chemoreceptive responses of two sympatric water snakes to extracts of commonly ingested prey species

Newborn of two locally sympatric species of water snakes,Nerodia fasciata andN. erythrogaster, were exposed to skin extracts of frequently ingested prey. In southern Louisiana, fish account for 78% of the diet of adultNerodia fasciata and about 15% forNerodia erythrogaster. Anurans comprise most of the remainder of the diets of these water snakes. Responses of naive individuals were compared to the chemoreceptive response profiles of groups of snakes reared on limited diets of fish and/or frogs. All snakes were tested seven times over a nine-month period and ontogenetic changes were apparent. NewbornNerodia fasciata have a strong chemoreceptive response for fish extract that remains unchanged by early dietary restrictions for the first six months of life. After that time their responses are more variable and can be altered by recent feeding experience. In southern Louisiana, this species is the least specialized water snake, an observation supported by these laboratory results. While newbornN. erythrogaster did not respond preferentially to any of the prey extracts, a significant response rate for fish extract is established by two months of age that persists for several months regardless of dietary restrictions. A subsequent shift of the response to frog extract at eight months of age regardless of diet may reflect a maturational process. Field studies are in agreement with these results and support the notion of ontogenetic changes related to predator size and age. This is the first demonstration of an ontogenetic change in chemoreceptive response that is not related to dietary experience.     

Antennal electrophysiological responses of three parasitic wasps to caterpillar-induced volatiles from maize (Zea mays mays), cotton (Gossypium herbaceum), and cowpea (Vigna unguiculata)

Many parasitic wasps are attracted to volatiles that are released by plants when attacked by potential hosts. The attractiveness of these semiochemicals from damaged plants has been demonstrated in many tritrophic systems, but the physiological mechanisms underlying the insect responses are poorly understood. We recorded the antennal perception by three parasitoids (Cotesia marginiventris, Microplitis rufiventris, and Campoletis sonorensis) to volatiles emitted by maize, cowpea, and cotton plants after attack by the common caterpillar pest Spodoptera littoralis. Gas chromatography-electroantennography (GC-EAG) recordings showed that wasps responded to many, but not all, of the compounds present at the physiologically relevant levels tested. Interestingly, some minor compounds, still unidentified, elicited strong responses from the wasps. These results indicate that wasps are able to detect many odorant compounds released by the plants. It remains to be determined how this information is processed and leads to the specific behavior of the parasitoids.     

Electroantennogram responses ofTrirhabda bacharides (weber) (Coleoptera: Chrysomelidae) to plant volatiles

Electroantennograms (EAGs) were recorded fromTrirhabda bacharides Le Conte (Coleoptera: Chrysomelidae) females to 28 odorants and volatiles emanating from leaves of 26 plant species including three known host plants,Baccharis halimifolia L.,B. neglecta Britt, andB. salicina T. & G. (Compositae: Asterae). Antennal receptors were highly responsive to components of the green leaf volatile complex, especially 6-carbon saturated and monounsaturated alcohols andtrans-2-hexenal. EAGs elicited by heptanal were greater than those elicited by any other saturated aldehyde. Oxygenated monoterpenes were more active than monoterpene hydrocarbons. -Bisabolol was the most active sesquiterpene. In general, EAGs to volatiles emanating fromBaccharis and other composite species were greater than those elicited by species from other families. Furthermore, when one considers only plants occurring in the environs ofT bacharides hostB. neglecta, EAGs elicited by its host plant were at least two times greater than those elicited by other coinhabiting plants.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.