Predation risk assessment by green frog (<Emphasis Type="Italic">Rana clamitans</Emphasis>) tadpoles through chemical cues produced by multiple prey

Many prey assess predation risk through predator chemical cues. Numerous studies have shown that (1) prey sometimes respond to chemical cues produced by heterospecifics and (2) that many species are capable of associative learning. This study extends this research by focusing on predation risk assessment and antipredator behavior in environments containing chemical cues produced by multiple prey species. The results show that green frog (Rana clamitans) tadpoles (1) assess risk from the chemical cue produced during predation by a heterospecific (gray tree frog, Hyla versicolor, tadpoles) and (2) can exhibit similarly strong behavioral responses to a mix of conspecific and heterospecific cues compared to conspecific cue alone, depending on their conditioning environment. I then discuss how the prey choice of the predators and the relative abundances of the prey species should influence the informational value of heterospecific cues.     

Benefit of suites of defensive behavior induced by predator chemical cues on anuran tadpoles, Hyla japonica

When predator chemical cues are present, low activity of prey is a commonly seen defensive behavior. However, few studies have explored the functional implications of the defensive behaviors and, thus, elucidated the possible linkages between behavioral responses and its consequences. In this study, we experimentally investigated how behavioral responses of Hyla japonica tadpoles to predator chemical cues affect vulnerability to a dragonfly nymph Anax parthenope julius. The frequency of tadpoles attacked by dragonfly nymphs was lower with chemical cues of predator was present than without chemical cues, and most of attacks occurred when tadpoles were mobile. When tadpoles were exposed to chemical cues, on the other hand, their swimming speed was quicker and swimming distance was longer, respectively, and the rates of being approached of the swimming tadpoles by dragonfly nymph was lower than those not exposed to chemical cues. We found that the tadpoles are induced by predator chemical cues not only to generally lower activity but also to swim in bursts as additional behavior and that the suite of their behavioral responses reduce the vulnerability against dragonfly nymph. Tadpoles can receive information about the predation risks by chemical cues and adjust their defensive behavior accordingly.     

Foods and predators of the green sea urchin Strongylocentrotus droebachiensis in Newfoundland waters

Gut analyses of the green sea urchin Strongylocentrotus droebachiensis (O. F. Müller) demonstrated that perennial phaeophytes, mostly fucoids and Alaria esculenta, were predominant in the diet. Ephemeral species, coralline algae and animals, were consumed in smaller amounts when available. Grazing by the urchins is evidently responsible for the dearth of non-coralline sublittoral algae in Newfoundland waters. Lobsters, rock crabs, purple sea stars, other urchins, and a variety of fishes and birds feed on S. droebachiensis, but predation is apparently not effective in limiting the abundance of the urchin.Studies in Biology from the Memorial University of Newfoundland No. 234.Contribution from the Marine Sciences Research Laboratory No. 66.     

Relative contribution of dietary carotenoids and vitamin E to visual and chemical sexual signals of male Iberian green lizards: an experimental test

Carotenoid-based sexual ornaments are widespread, but the role of carotenoids as honest signalers in the trade-off between coloration and antioxidant protection remains controversial. It has been suggested that the function of carotenoids might not be an antioxidant per se, but that colorful carotenoids may indirectly reflect the levels of nonpigmentary antioxidants, such as melatonin or vitamin E. We experimentally fed male Iberian green lizards (Lacerta schreiberi) additional carotenoids or vitamin E alone, or a combination of carotenoids and vitamin E dissolved in soybean oil, whereas a control group only received soybean oil. We examined the effects of the dietary supplementations on characteristics of lizard sexual coloration and of the chemical profile of femoral gland secretions. Results indicated that both carotenoids and vitamin E in the diet increased the expression of some visual signals (head and chest, but not dorsum) in comparison to controls. However, different traits were differentially affected, and in many cases, the addition of vitamin E, alone or in combination with carotenoids, had a greater effect on the expression of coloration than the addition of carotenoids alone, even for carotenoid-dependent ornaments. Our results support the idea that other nonpigmentary antioxidants, such as vitamin E, are needed in addition to carotenoids to increase the expression of coloration of L. schreiberi lizards. Therefore, coloration may only indirectly reflect the levels of nonpigmentary antioxidants. In contrast, an increase in dietary nonpigmentary antioxidant vitamin E was directly reflected in the chemical signals. Because of an observed concordance between visual and chemical signals, we suggest that both may be used in different contexts albeit conveying similar messages in different sensory channels.     

Use of chemical cues in detection of conspecific predators and prey by newts, <Emphasis Type="Italic">Notophthalmus viridescens</Emphasis>

Summary. Detection of heterospecific predators and prey via chemical cues is well known, but only a few studies have examined the potential for such discrimination in cannibalistic systems. In newts, Notophthalmus viridescens, adults are opportunistic predators of conspecific larvae. I used a laboratory bioassay to determine whether larvae and adults distinguish between chemical stimuli from members of the different age classes. Larvae distinguished between chemical stimuli from larvae and adults by decreasing their activity only following exposure to stimuli from adults. Decreased activity is consistent with an antipredator response in many prey species, including larval newts. In contrast, adults increased their activity and increased time spent in open areas in response to stimuli from larvae, but not to stimuli from adults. Increased activity is consistent with a feeding response; adults also showed increased activity and increased time in open areas in response to chemical stimuli from familiar heterospecific prey (brine shrimp). The proximate cue that allowed the newts to distinguish between the different age classes is not associated with short-term dietary differences because all stimulus animals were fed the same prey. Stimulus strength was controlled by diluting the stimulus solutions according to the volume of the stimulus animal. Therefore, there appear to be intrinsic differences in the chemical signatures of larval and adult newts.     

Hierarchical perception of fertility signals and nestmate recognition cues in two dolichoderine ants

In social insects, queens are likely to “honestly” inform their nestmates of their presence and fertility status through pheromonal communication. Cuticular hydrocarbons (CHCs) have been reported to be effective nestmate discriminators and strongly suspected to act as fertility signals, at least in some species. The use of the same chemical bouquet (i.e., the CHC profile) to convey two fundamentally different information seems puzzling. However, a recent threshold model proposes a hierarchy in the discriminating processes, i.e., fertility signals can only be perceived if nestmate recognition has been reached (Le Conte and Hefetz, Annu Rev Entomol 53:523–542, 2008). Here, we developed a simple behavioral bioassay based on chemical recruitment toward a queen placed outside the nest in two dolichoderine ants (Linepithema humile and Tapinoma erraticum), which allowed us to investigate the interplay between fertility signaling and colonial recognition. Using queen corpses of various origins (nestmates or aliens) and physiological states (fertile or infertile; mated or unmated), we demonstrated that nestmate recognition cues clearly override fertility signals under our experimental conditions. Indeed, while nestmate infertile queens were largely ignored by the workers, nestmate fertile queens (mated or not) induced worker recruitment, whereas alien fertile queens did not and were aggressed by the workers.     

Social recognition in sticklebacks: the role of direct experience and habitat cues

The social organisation of animals relies on recognition. However, there are many means by which animals may recognise one another and a variety of cues are available to any individual at any one time. We tested the effects of cues based on direct experience and of cues based on habitat and diet on association decisions in the three-spined stickleback (Gasterosteus aculeatus). Our results show that sticklebacks rapidly acquire association preferences for novel con- and heterospecifics (nine-spined sticklebacks, Pungitius pungitius) which had experienced the same habitat and diet conditioning as themselves over novel con- and heterospecifics which had experienced a different habitat and diet conditioning, a preference which may be based on self-referent matching. Association preferences were observed after only 24 h and were independent of the number of tank mates (treatments with 20 and 100 fish). Evidence for the influence of direct social experience on association preferences was weak and may be confounded by other factors.Communicated by K. Lindström     

Ontogeny of nestmate recognition cues in the red carpenter ant (Camponotus floridanus)

Summary A combination of behavioral and chemical analyses was used to investigate the nature of nestmate recognition cues and the effects of worker age and social experience on these cues in the ant Camponotus floridanus. Five categories of workers were tested: foragers, 5-day old and 0-day old callows, 5-day old and 0-day old naive callows. Bioassays consisted of introductions of dead workers from these categories into their own colonies or into an alien colony after the following treatments: 1) killed by freezing, 2) solvent-washed, 3) solvent-washed and coated with a nestmate soak, 4) solvent-washed and coated with a non-nestmate soak. Soaks were obtained from individual ants immersed in hexane and were applied individually to washed workers from the same category. Soaks were analyzed by gas chromatography (GC) and compared by multivariate analyses. Freeze-killed workers from each category elicited more aggressive behavior in the alien colony than in its own. By comparing GC profiles, a worker from any category can be assigned to its colony of origin. Thus all studied worker categories are colony-specific. Solvent-washed ants did not induce more aggressive behaviors in the alien colony than in their own, but they induced significantly less aggressivity in an alien colony than non-treated dead ants from the same category. Washed ants indced more aggressive behaviors when coated with a soak from a different colony as opposed to a soak from the colony in which they were introduced. The combination of behavioral and chemical results lead to the following conclusions: 1) Information contained in soak derived from workers was sufficient to allow nestmate recognition. 2) Nestmate recognition cues, and consequently the recognition response displayed to their bearer, change with age. 3) Social experience is necessary to develop or acquire a colony-specific label. The role of age and social experience on nestmate recognition in social Hymenoptera is discussed.     

The effect of hunger on the strength and duration of the antipredator behavioral response of green frog (<Emphasis Type="Italic">Rana clamitans</Emphasis>) tadpoles

The activity level of prey reflects a trade-off between predation risk and foraging gain. A number of theoretical and empirical studies have shown that a prey's energetic state or the level of its resource should influence this trade-off (i.e., what the optimal activity level at a level of predation risk is). Here, I show that the energetic state of prey may also influence the duration of their antipredator behavioral response. Green frog tadpoles (Rana clamitans) reduced their activity level for a shorter time during exposure to the chemical cue of predatory larval dragonflies (Anax spp.) as their time since last feeding increased (i.e., as their energetic state decreased). Interestingly, the tadpoles strongly reduced their activity level upon cue exposure in all treatments. Thus, the relative activity level of tadpoles at different energetic states varied over time.     

Herbicide exposure affects the chemical recognition of a non native predator in common toad tadpoles (<Emphasis Type="Italic">Bufo bufo</Emphasis>)

Summary. In amphibians and fishes, evidence is increasing that chemical cues from injured conspecifics can play a role in the chemical labelling and learned recognition of unfamiliar predators. In this laboratory study, we tested the prediction that prior chemical exposure to a non-native predator feeding on conspecific tadpoles will subsequently allow tadpoles of the common toad (Bufo bufo) to recognize the chemical cues specifically released by this starved predator. Furthermore, we investigated the vulnerability of this chemically-mediated process to herbicide contamination. With these aims in view, groups of tadpoles were kept either unexposed or exposed for ten days to chemical cues from Turkish crayfish (Astacus leptodactylus) previously fed on tadpoles, both in uncontaminated water and in the presence of four sublethal concentrations of amitrole (0.01, 0.1, 1 and 10 mg.l⁻¹). We then assessed the effects of the six conditioning treatments on general activity and behavioural response to chemical cues from starved crayfish. Larval treatments did not affect the general activity of the tadpoles. By contrast, the treatments had significant effects on the behavioural response to the test solution prepared form starved crayfish. The only tadpoles to show an antipredator behavioural response to the chemical stimulation from starved crayfish belonged to the groups derived from chemical exposure to tadpole-fed crayfish in uncontaminated water and in contaminated water with the lowest concentration of amitrole (0.01 mg.l⁻¹). Conversely, this chemical stimulation produced no behavioural change in the control group or in the groups derived from exposure to tadpole-fed crayfish in contaminated water containing 0.1, 1 and 10 mg.l⁻¹ of amitrole. This study demonstrates that chemical cues released during the predator’s feeding activity can subsequently be used by common toad tadpoles in the recognition of an unfamiliar predator. In addition, our results show that the presence of sublethal amitrole concentrations can impair this recognition process. Such a pesticide effect might be especially detrimental for amphibian populations threatened by invasive predators.     

Adaptive behaviour of chironomid larvae (Chironomus riparius) in response to chemical stimuli from predators and resource density

Chemical-mediated effects of predatory fish on chironomid larvae behaviour have been ignored so far. Sediment-dwelling chironomid larvae inhabit protective burrows from which they extend their bodies only to feed on deposited detritus and microalgae from the surrounding sediment. Here, we performed factorial laboratory experiments to study whether fish-borne chemical cues (kairomones) are responsible for behavioural trait changes of chironomid larvae, and whether chironomid larvae are able to assess the densities of fish predators and food resources and the trade-off between them. We exposed naïve Chironomus riparius larvae to the chemical presence of zero, one, and ten predator fish (Rutilus rutilus) and offered two resource levels (low food, high food) for each treatment. Kairomones induced significant inherent behavioural trait changes in chironomid larvae. During the first 120 min after exposing chironomids to fish-conditioned water, we found a significant increase in digging activity with increasing predator density. After 3 days of exposure, the deepest chironomid burrows were found in treatments with the highest predator density. Chironomid larvae were significantly able to adjust their foraging behaviour to different predator densities and food concentrations and trade off between them; that is, when fish predators were more abundant or when more food resources were available, the foraging activities of larvae were significantly reduced. Our data suggest that chemically mediated trait changes (burrowing and foraging behaviour) may cascade through the littoral food web.     

Effects of embryonic exposure to conspecific chemical cues on hatching and larval traits in the common frog <Emphasis Type="Italic">(Rana temporaria)</Emphasis>

Summary. Recent studies indicate that amphibian eggs are capable of hatching plasticity in response to chemical cues released by predators feeding on conspecific eggs or larvae. However, information is scarce on the relative importance of predator and conspecific cues in such a process. In particular, no attempt has been made to compare the effects of embryonic exposures to chemical cues indicative of a predation risk for eggs and larvae, although both life stages can co-occur in natural habitats. In this context, common frog embryos (Rana temporaria) were raised until hatching in the presence of crushed conspecific extracts from eggs and tadpoles to assess their respective influences on some hatching and larval traits. While a significant delay in hatching time was observed in embryos exposed to chemical cues from tadpole extract, this life-history shift appeared unaffected by embryonic exposure to egg extract. Hatchlings derived from eggs incubated in the presence of both conspecific extracts showed a significantly greater weight than unexposed controls. However, such an effect was no longer apparent 15, 30 and 50 days after hatching, suggesting that embryonic exposure to chemical cues from damaged conspecific eggs and tadpoles has no influence on larval growth. Lastly, morphological measurements performed on hatchlings and older tadpoles (15, 30 and 50 days old) revealed no significant effect of embryonic treatments on the shape of body and tail.     

Transmission distance of chemical cues from coral habitats: implications for marine larval settlement in context of reef degradation

The present study (Ishigaki Island, Japan) explored the distance of transmission of chemical cues emitted by live versus dead coral reefs (Exp. 1: High performance liquid chromatography (HPLC) analyses with water sampling station at 0, 1, and 2 km away from the reef) and the potential attraction of these chemical cues by larval fish, crustaceans, and cephalopods (Exp. 2: choice flume experiment conducted on 54 Chromis viridis larvae, 52 Palaemonidae sp larvae, and 16 Sepia latimanus larvae). In the experiment 1, HPLC analyses highlighted that the live coral reef (and not the dead coral reef) produced different and distinct molecules, and some of these molecules could be transported to a distance of at least 2 km from the reef with a reduction of concentration by 14–17-fold. In the experiment 2, C. viridis, Palaemonidae sp, and S. latimanus larvae were significantly attracted by chemical cues from a live coral reef (sampling station: 0 km), but not from a dead coral reef. However, only C. viridis larvae detected the chemical cues until 1 km away from the live coral reef. Overall, our study showed that chemical cues emitted by a live coral reef were transported farthest away in the ocean (at least 2 km) compared to those from a dead coral reef and that fish larvae could detect these cues until 1 km. These results support the assumption of a larval settlement ineffective in degraded coral reefs, which will assist conservationists and reef managers concerned with maintaining biodiversity on reefs that are becoming increasingly degraded.     

Predator-induced alarm responses in the common periwinkle, Littorina littorea: dependence on season, light conditions, and chemical labelling of predators

Chemically mediated alarm reactions of the common periwinkle, Littorina littorea (L.), were studied in laboratory experiments during two consecutive summers, and one intermediate autumn season. Responses to chemical stimuli were detected as crawl-out responses, i.e. movements of snails out of the water. Snails were exposed to extracts of injured conspecifics, extracts of the mussel Modiolus modiolus (L.), and water conditioned by the predatory crab Carcinus maenas (L.), which had been maintained on different diets. In experiments carried out during the summer, a significantly larger number of snails moved out of the water when exposed to chemical stimuli from injured conspecifics, compared to chemical stimuli from injured mussels or filtered seawater. These results suggest that chemical alarm substances are present in L. littorea. Water conditioned by crabs that had been fed L. littorea released significantly more crawl-out responses compared to water conditioned by crabs that had been kept on a fish diet. When tested in autumn, no significant differences were found in responses to the above-mentioned water samples. Crawl-out responses under different light regimes were also investigated. All series of experiments carried out in the dark evoked a higher number of responses compared to series that took place in light. These findings may indicate an adaptation of snails to night-active predators. In total, the current results suggest that a L. littorea diet may chemically “label” the predator crab with snail alarm substances, and that predator-induced responses of L. littorea are actually responses to conspecific alarm substances released from crabs that have been maintained on a L. littorea diet. The response to the alarm signal, however, appears to be dependent on season and light conditions; some ecological implications of these findings are also discussed. Received: 8 January 1999 / Accepted: 29 March 1999     

Induction of host specificity in larvae of Manduca sexta: chemical dependence controlling host recognition and developmental rate

Summary. The tobacco hornworm, Manduca sexta L. (Lepidoptera: Sphingidae) is a specialist on Solanaceae. This host specificity is induced as the larva feeds on solanaceous foliage, so that solanaceous-reared larvae will refuse to feed on other plants. Experiments were designed to determine the role of dietary constituents on the induction of host specificity and the effects of these on development of M. sexta. Choice assays using leaf discs of cowpea, Vigna sinensis (Fabaceae), were used to monitor the isolation of relevant chemical cues from foliage of potato. An aqueous extract of potato foliage strongly stimulated feeding. This extract was partitioned with n-butanol under alkaline conditions to obtain a highly active butanol extract. Reversed phase flash chromatography with a water-methanol gradient gave an active fraction that was used as a supplement for wheat germ-based artificial diet. Larvae reared on this s-diet became dependent on potato allelochemicals for initiation and continuation of feeding activity. These larvae also developed faster than larvae reared on the control p-diet, but no effect on adult mass was detected. Further flash chromatography of the active fraction under alkaline conditions provided a highly active sub-fraction, and semi-preparative HPLC using gradients of water and acetonitrile resulted in the isolation of a single compound. Bioassays indicated that this compound alone can account for host recognition by solanaceous-reared larvae. The results suggest that the mechanism of induced host specificity in M. sexta involves development of dependence on this compound. Received 21 December 1999; accepted 14 March 2000     

Synthesis of nickel nanoparticles by chemical and green route and their comparison in respect to biological effect and toxicity

Recent reports demonstrated an increase in the synthesis of nanoparticles (NP) and its applications. The chemical preparation of NP may be harmful. Thus, in this study, the biological activity and toxicology of nickel nanoparticles (NiNP) synthesized through chemical and green routes were compared. Chemical synthesis of NiNP was mediated by polyethylene glycol and hydrazine hydrate as stabilizing and reducing agent, respectively. Desmodium gangeticum aqueous root extract was used to prepare NiNP without any stabilizing and reducing agent. Nickel nanoparticles synthesized by both methods were characterized (ultraviolet–visible spectroscopy, X-ray diffractrometry, Fourier transform infrared spectroscopy, zeta potential and vibrating sample magnetometer) and compared. There was no major significant difference in the nature of the NP prepared by both methods. However, green synthesized NiNP showed reduced size and better monodispersity compared to chemical synthesized one. Free radical scavenging activities and antibacterial activities of NiNP prepared by chemical and green route suggest that these NP, prepared by green route possess reliable antioxidant and antibacterial activity. Evaluation of toxicity in animal and cell line suggests that NiNP synthesized by green route are nontoxic.     

Control of larval release in the Caribbean spiny lobster, <Emphasis Type="Italic">Panulirus argus</Emphasis>: role of chemical cues

The current model for larval release in subtidal crustaceans suggests that hatching time is controlled by the embryos, which release a pheromone that stimulates the parent female to undergo behaviors that synchronize larval release. Alternatively, hatching could be controlled by the females. Ovigerous spiny lobsters Panulirus argus (Latreille) exhibit stereotypic behaviors during larval release, including rapid abdominal extensions and pleopod-pumping activity. Ovigerous P. argus were collected from coral reefs in the Florida Keys, USA during the summers of 2005 and 2006. Pleopod-pumping activity was quantified to determine if a female’s pumping activity correlates with the developmental state of the embryos. The role of pheromones released by developing and hatching embryos in controlling pumping behaviors was tested by measuring the pumping response of ovigerous lobsters to (1) hatch water, (2) homogenized embryo water, (3) embryo-conditioned water (unhatched late-stage embryos soaked for 20 h), and (4) water containing homogenized post-hatch embryo cases. Bioassays were conducted under constant conditions (dim-red light) in the laboratory at random times during the day to control for any possible rhythm in pumping activity. Spontaneous pleopod-pumping activity increased significantly with increasing embryo development. Upon exposure to hatch water, ovigerous lobsters with late-stage embryos displayed increased pleopod pumping with increased treatment concentration. Water individually conditioned with homogenized late-stage embryos, intact late-stage embryos, and homogenized post-hatch embryo cases all induced larval release behaviors in females with late-stage embryos. Ovigerous females with early-stage embryos did not respond to water conditioned with homogenized early- or late-stage embryos. These results suggest that active substances are released by embryos at the time of hatching and induce the stereotypical pumping behaviors of the female that synchronizes larval release. The results support the model that larval release in subtidal crustaceans is controlled by pheromones released from hatching embryos.     

The meta-hydroxylation step in the phenylpropanoid pathway: a new level of complexity in the pathway and its regulation

Together in silico and genetic mining approaches have recently designated the CYP98 family of plant cytochromes P450 as the family of enzymes that catalyzes the meta-hydroxylation step in the phenylpropanoid pathway. This meta-hydroxylation is not catalyzed on the free p-coumaric acid as anticipated, but on its conjugates with shikimic, quinic, or phenyllactic acids. While all CYP98s have in common phenol meta-hydroxylase activity, p-coumaroylshikimate remains their preferred substrate. High expression of CYP98s is detected in lignifying tissues in stems, roots, and siliques. The CYP98A3 gene disruption in Arabidopsis thaliana leads to a drastic inhibition of lignin synthesis, cell growth, and plant development. The meta-hydroxylation of phenolic precursors is thus essential for higher plant development. Isolation of coding sequences belonging to the CYP98 family from basil, wheat, and extensive functional analysis of the recombinant enzymes, together with CYP98s from other plant taxa, helps shedding some light on mechanisms of P450s evolution. Most importantly, the occurrence of the meta-hydroxylation on esters of shikimic or quinic acids introduces a new biochemical regulation mechanism in the phenylpropanoid pathway.     

Interpopulational differences in chemical composition and chemosensory recognition of femoral gland secretions of male lizards Podarcis hispanica: implications for sexual isolation in a species complex

Summary. Molecular studies suggest that the Iberian wall lizard, Podarcis hispanica, forms a species complex with several monophyletic types. In Central Spain two of these types are spatially not isolated and may interact. Sex pheromones are important for species recognition and, thus, differences between lizards’ types in chemicals used in intraspecific communication could lead to reproductive isolation. Analyses by GC-MS showed that the femoral gland secretions of adult males of different types were different. Males of one type had twelve exclusive compounds, and proportions of some shared compounds differed. This presumably would reflect selection for the persistency and efficiency of chemical signals in different environments; less volatile compounds and with a higher chemical stability being favoured in the type of lizards inhabiting more humid climatic conditions. Differential tongue-flick rates to scents from femoral secretions indicated that males were able to detect and discriminate between males of different types based on chemical cues alone. In contrast, females detected but did not seem able to discriminate between scents of the two types of males. Thus, multiple factors might be simultaneously acting either against or in favor of speciation, leading, on the one hand, to genetic differences between types, but, on the other hand, probably precluding an effective reproductive isolation in areas where both types of lizards may interact.