Response to prey chemical cues by hatchling pine snakes (Pituophis melanoleucus): Effects of incubation temperature and experience

The ability of hatchling pine snakes (Pituophis melanoleucus) to select and follow or avoid chemical odors of prey (mice,Mus musculus) on a shavings and paper substrate was investigated in Y-maze experiments, as a function of incubation temperature and experience. Incubation temperature affected behavior in the maze, and the maze choices of naive snakes, but not of snakes that had already eaten a mouse. The data indicate that snakes that have eaten, preferentially enter the arm bearing chemical stimuli from mice, whereas those that have not eaten show no preference.     

Effect of experience with pine (Pituophis melanoleucus) and king (Lampropeltis getulus) snake odors on Y-maze behavior of pine snake hatchlings

The abilities of hatchling pine snakes (Pituophis melanoleucus) and king snakes (Lampropeltis getulus) to discriminate the chemical trails of pine and king snakes was investigated inY-maze experiments. Pine snakes were housed for 17 days either with shavings impregnated with pine snake odor, king snake odor, or no odor to test for the effect of experience on choice. Both pine and king snake hatchlings entered the arm with the pine snake odor and did not enter the arm with the king snake odor. The data support the hypothesis that hatchlings of both species can distinguish conspecific odors from other odors and that our manipulation of previous experience was without effect for pine snake hatchlings.     

Effect of conspecific and heterospecific feces on foraging and oviposition of two predatory ladybirds: role of fecal cues in predator avoidance

Growing evidence suggests a flow of chemical information from higher to lower trophic levels that affects foraging and oviposition of prey in response to potential risks from predators. This was investigated in two species of ladybird predators of aphids, Harmonia axyridis and Propylea japonica. H. axyridis is known to be the stronger intraguild predator and P. japonica to be the more frequent intraguild prey in interactions of these two species. These ladybirds share aphid prey on mugworts, hibiscus, and Italian ryegrasses in fields of northern Japan but largely avoid each other on the same plant. Fecal cues of these ladybird predators were found to contribute in their assessment of predation risk from conspecific and heterospecific competitors in common habitats. Gravid females of H. axyridis reduced rates of feeding and oviposition when exposed to feces of conspecifics, but not when exposed to feces of P. japonica. In contrast, gravid females of P. japonica reduced feeding and oviposition when exposed to feces of both H. axyridis and its own species. Females of both ladybird species exhibited similar behavior in response to water extracts of feces. For P. japonica, the influence of heterospecific feces was greater than that of conspecific feces. Our results demonstrate that feces of ladybirds contain odors that have the potential to deter the feeding and oviposition activities of conspecific as well as heterospecific ladybirds. Such deterrence allows these insects to avoid predation risk. Differences in responses of the two predators are discussed.     

Naive ophiophagus lizards recognize and avoid venomous snakes using chemical cues

Monitor lizards prey on snakes. Conversely, venomous snakes prey on juvenile monitor lizards. Immediately after hatching, monitor lizards are naive to all prey items, thus correct assessment of snake prey is paramount for survival. Experiments were conducted to determine how hatchling monitor lizards (Varanus albigularis) with no previous exposure to snakes reacted to sympatric venomous and nonvenomous snakes. Hatchling lizards attacked harmless snakes, but avoided venomous species. Lizards readily accepted meat from skinned snakes, regardless of species. When invertebrate prey covered with skin segments from venomous snakes were restrained from moving, they were usually investigated by tongue-flicking and rejected. Unrestrained skin-covered prey, however, were generally attacked and eaten without prior evaluation by tongue-flicking. Attack was inhibited in trials in which unrestrained prey were tongue-flicked, suggesting that chemical cues contained in snake skins mediate avoidance of venomous snakes. Selection for the ability to perceive snake integumental chemicals may be especially strong in species that both consume and are consumed by snakes.     

Responses by corn snakes (Elaphe guttata) to chemicals from heterospecific snakes

Young corn snakes,Elaphe guttata, were tested for responses to chemicals from heterospecific snakes. Corn snakes exhibited more tongue-flicks to swabs freshly rubbed against the skin of an ophiophagous kingsnake,Lampropeltis getulus, than to blank swabs. Responses toL. getulus and a nonophiophagous western plains garter snake,Thamnophis radix haydeni, did not differ significantly. Corn snakes exhibited more tongue-flicks to swabs treated with chloroform extracts of the shed skins ofL. getulus; an ophiophagous eastern coachwhip,Masticophis flagellum; and a nonophiophagous gray ratsnake,Elaphe obsoleta, than to blank swabs, but they did not discriminate between ophiophagous and nonophiophagous species in every case. Corn snakes, when offered shelters containing bedding from the home cages of a nonophiophagous water snake,Nerodia erythrogaster, an occasionally ophiophagous water moccasin,Agkistrodon pisdvorus; orL. getulus and untreated bedding, failed to reside under snake-scented shelters at a rate significantly different from that expected by chance. The responses of corn snakes are compared with those reported for other snakes presented with heterospecific snake chemicals.     

Avoidance of conspecific injury-released chemical cues by free-ranging Gammarus lacustris (Crustacea:Amphipoda)

Behavioral responses to chemical cues have been demonstrated for a range of aquatic animals. Injury-released chemical alarm cues from conspecifics are released when a prey's predator is actively foraging. Detection of these cues elicits antipredator behaviors that reduce the probability of predation. Amphipod crustaceans in the genus Gammarus are widespread denizens of ponds and streams. Antipredator responses by Gammarus to conspecific alarm cues, and subsequent reduction of predation risk, are known from experiments in the laboratory. Here, we verify laboratory findings by demonstrating an avoidance response to alarm cues using a field population of G. lacustris. We used small traps baited with sponge blocks containing either water (control) or injury-released cues from Gammarus. We repeated the experiment twice. In both experiments, significantly fewer Gammarus were captured in traps with alarm cue sponges than in traps with water sponges. Predatory leeches Dina parva were attracted to Gammarus traps in the first experiment but not the second experiment. In the second experiment, we measured the individual weight of captured amphipods. Two size classes were present; small (1-5 mg) and large (35-108 mg). Both sizes contributed to the avoidance response. Within the large size class, small individuals were significantly less responsive to the alarm cue than large individuals, implying that small adult Gammarus may have different cost/benefit decision criteria for risk assessment than large Gammarus.     

Avoidance response of a terrestrial salamander (Ambystoma macrodactylum) to chemical alarm cues

Organisms from a wide variety of taxonomic groups possess chemical alarm cues that are important in mediating predator avoidance. However, little is known about the presence of such alarm cues in most amphibians, and in particular terrestrial salamanders. In this study we tested whether adult long-toed salamanders (Ambystoma macrodactylum) showed an avoidance response to stimuli from injured conspecifics. Avoidance of stimuli from injured conspecifics could represent avoidance of a chemical alarm cue or, alternatively, avoidance of a territorial pheromone or conspecific predator odor. Consequently, we also tested whether salamanders avoided stimuli from noninjured conspecifics. Salamanders avoided stimuli from injured but not from noninjured conspecifics. Therefore, we concluded that the response to injured conspecifics represents avoidance of a chemical alarm cue and not avoidance of a territorial pheromone or predator cue. This is the first clear demonstration of chemical alarm signaling by a terrestrial amphibian and the first report of chemical alarm signaling in an ambystomatid salamander. By avoiding an area containing stimuli from injured conspecifics, long-toed salamanders may lower their risk of predation by avoiding areas where predators are foraging.     

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.     

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.     

Chemicals of predatory mosquitofish (Gambusia affinis) influence selection of oviposition site by Culex mosquitoes

Ovipositing insects may avoid aquatic sites where there is high predation risk to their offspring, but the proximate mechanisms that mediate avoidance behavior are poorly resolved. We conducted an experiment to determine whether mosquitoes would reduce oviposition rates in pools containing chemicals of the mosquitofish (Gambusia affinis), a voracious predator that is widely employed to control mosquitoes. Experimental treatments consisted of outdoor pools that contained known concentrations of fish chemicals (low, medium, or high) or no fish chemicals (control). The pools were arranged in a randomized block design, and the number of mosquito larvae in each pool served as the response variable to estimate relative oviposition rate. Members of the Culex pipiens complex were the main colonizers of the pools. The mean number of larvae per pool differed among treatments (P = 0.026) and was about three times greater in control pools compared with those receiving medium and high concentrations of fish chemicals. Pairwise comparisons indicate that only medium and high treatments differed significantly from controls, suggesting that a threshold concentration exists below which mosquitoes cannot reliably detect predators. Our data suggest that the effectiveness of Gambusia affinis in controlling mosquitoes may be compromised if adult mosquitoes respond to fish stocking by shifting to nearby breeding sites that lack fish. We discuss issues concerning the use of Gambusia in biological control programs within the context of these new findings.     

Purification and preliminary characterization of a frog-derived proteinaceous chemoattractant eliciting prey attack by checkered garter snakes (Thamnophis marcianus)

A potent proteinaceous chemoattractant, eliciting prey attack by checkered garter snakes (Thamnophis marcianus) was isolated from aqueous washes of the common frogRana temporaria and purified by preparative continuous-elution electrophoresis. The biological activity of the frog crude extract or of the purified chemoattractive protein, measured by a snake bioassay, was unaffected by freezing, lyophilization, or dialysis but was lost after proteolytic digestion. The purified chemoattractant is glycosylated, has an apparent molecular mass of 24 kDa, estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE), and a pI of 4.8. It gave one spot in two-dimensional electrophoresis. The bioassay showed that this protein is highly attractive to snakes. The lowest concentration yielding positive responses in the snake bioassay was approximately 25 µg/ml. These results suggest that a water-soluble Mr 24 kDa glycoprotein molecule produced by the common frog may be a vomeronasal stimulus used by checkered garter snakes for prey recognition.     

Responses to inter- and intraspecific scent marks in pine martens (Martes martes)

In order to analyze intra and interspecific olfactory discrimination, behavioral responses (sniffing and marking) towards various odors were observed in pine martens (Manes martes). Two adult males and two adult females were tested for intraspecific discrimination of abdominal gland odors and urine. Both sexes sniffed and marked objects carrying their own scent less than unscented objects. There were no differences in sniffing or marking objects impregnated with odors from known and unknown conspecifics of the opposite sex or objects carrying the odor of an unknown male or female. A second experiment with two adult females gave no evidence for interspecific discrimination: no differences emerged when comparing reactions towards marks of pine martens, stone martens (a closely related species), or genets. The most obvious result of this study is the reduced response of pine martens towards their own mark. It is suggested that scent marking in martens may reflect autocommunication, the primary effect being to familiarize an animal with its environment.     

Responses of beaver (Castor canadensis Kuhl) to predator chemicals

Free-ranging beaver (Castor canadensis) in two different beaver populations in New York State were exposed to predator chemicals to test feeding inhibition. Solvent extracts of feces were applied to stem sections of aspen, the preferred food tree of beavers, permitting smelling and tasting the samples. Predator odors were from wolf (Canis lupus), coyote (Canis latrans), dog (Canis familiaris), black bear (Ursus americanus), river otter (Lutra canadensis), lynx (Lynx canadensis), and African lion (Panthera leo). The experiment was repeated. The predator odors reduced feeding compared to untreated or solvent-treated controls. One population consumed 17.0% of the samples with predator odor and 27.0% of the controls in summer, and 48.4% and 60.0%, respectively, in autumn. The other population accepted 3.15% of the predator odor samples and 11.05% of the controls in summer. Coyote, lynx, and river otter odors had the strongest effects. Diesel oil and bitter-tasting neem extract had weaker effects. Predator odors are promising as feeding repellents for beaver.     

Comparison of earthworm- and fish-derived chemicals eliciting prey attack by garter snakes (Thamnophis)

Materials eliciting increased tongue flicking and prey attack in garter snakes were isolated from both earthworm and fish prey. New extraction methods based on chloroform-methanol mixtures are valuable adjuncts to the more typical aqueous preparations. Both high- and low-molecular weight components from earthworms and fish were active. The similarity between the active chemicals in these two classes of prey was established using several methods of analysis. These included chromatography, carbohydrate and amino acid analyses, and nuclear magnetic resonance.     

Attraction of pinyon pine bark beetle,Ips hoppingi,to conspecific andI. confusus pheromones (Coleoptera: Scolytidae)

Females of a pinyon pine bark beetle,Ips hoppingi Lanier, were less attracted by the aggregation pheromone produced by conspecific males than by the pheromone produced by the neighboring sibling species,I. confusus (LeConte). Cross-attraction was elicited by males infesting the regional pinyon pine hosts (P. discolor andP. edulis) of eitherIps species in south-eastern Arizona. Pheromonal specificity has not accompanied speciation in this species pair.     

Chemical cues used by prairie rattlesnakes (Crotalus viridis) to follow trails of rodent prey

Each of 10 prairie rattlesnakes (Crotalus viridis) was exposed to three types of trails after striking rodent prey (Mus musculus). One trail was made with mouse urine, another was made with tap water, and the third consisted of materials from mouse integument. The snakes exhibited trailing behavior only when integumentary trails were available. It was concluded that prairie rattlesnakes do not utilize urinary cues; instead they attend to materials associated with rodent skin and fur.     

Responses of Wild Norway Rats (Rattus norvegicus) to Predator Odors

If rats could be shown to avoid the odors of predators, then conservation managers could manipulate this behavior to exclude rats from important conservation sites. We evaluated the ability of six predator odors to elicit avoidance responses by wild-caught Norway rats (Rattus norvegicus) from two New Zealand populations (Kapiti Island and North Island). Kapiti Island is free of mammalian predators, while the North Island has established populations of felids, canids, and mustelids. Three of the predator odors were real and three were synthesized volatile ingredients of real animal feces or urine. We compared the rats' responses to predator odors with their responses to three natural herbivore odors. We used a Y maze, and rats were offered the choice of an odor in one arm of the maze and distilled water in the other arm. Each choice arm was ducted at the entrance to remove air and the odor. We recorded: (1) the time until the rat left the first arm of the maze, (2) the time until the rat visited each arm, (3) the number of visits to each arm, (4) the amount of time spent in each arm, and (5) a total activity score for each rat. Kapiti rats showed an aversion to five of the six predator odors, despite never having encountered them before. Kapiti rats visited herbivore odors more often than carnivore odors and were less active in the presence of carnivore odors than they were when tested with herbivore odors. In addition, Kapiti rats approached some herbivore odors more quickly than they approached carnivore odors. North Island rats appeared to avoid cat urine, but despite being predator experienced, did not show a consistent aversion to the carnivore odors we tested. Both samples of rats, but particularly the North Island group, showed high individual variation. We conclude that predator odors are unlikely to be an effective management tool for all populations of this species of rat because of this high individual variability and the likelihood that each island population will differ in its responses to a given odor.     

Analysis of chemicals from earthworms and fish that elicit prey attack by ingestively naive garter snakes (Thamnophis)

Materials previously shown to elicit increased tongue-flicking and prey attack in garter snakes (Thamnophis sirtalis) were isolated from both earthworms (Lumbricus terrestris) and fish (Pimephales promelas). Both high- and low-molecular-weight components from earthworms and fish stimulated attacks and increased tongue-flicking in previously unfed neonate garter snakes relative to distilled water controls. Earthworm collagen was also effective, but even concentrated fractions were less effective than raw extract. Conflicting reports on the effectiveness of collagen suggest that the salient chemical(s) is a smaller molecule tightly bound to collagen and resisting standard purification methods.     

Response of brown tree snakes (Boiga irregularis) to human blood

Ten specimens ofBoiga irregularis were presented with clean or bloody tampons. The latter were used by women during menses. Trial duration was 60 sec, intertrial interval was 24 hr, and the dependent variable was rate of tongue flicking (a measure of chemosensory investigation). Bloody tampons elicited significantly more tongue flicking than did control tampons. An additional snake is shown attacking and ingesting a soiled tampon, confirming that chemosensory interest was associated with predatory behavior.     

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.