Behavioural responses to con- and heterospecific alarm cues by an alien and a coexisting native fish

Hydrobiologia - The monkey goby Neogobius fluviatilis is an invasive Ponto-Caspian fish that enters habitats of the native gudgeon Gobio gobio in European freshwaters, likely belonging to the same...     

Effects of degraded optical conditions on behavioural responses to alarm cues in a freshwater fish

Prey organisms often use multiple sensory cues to gain reliable information about imminent predation threat. In this study we test if a freshwater fish increases the reliance on supplementary cues when the reliability of the primary cue is reduced. Fish commonly use vision to evaluate predation threat, but may also use chemical cues from predators or injured conspecifics. Environmental changes, such as increasing turbidity or water colour, may compromise the use of vision through changes in the optical properties of water. In an experiment we tested if changes in optical conditions have any effects on how crucian carp respond to chemical predator cues. In turbidity treatments we added either clay or algae, and in a brown water colour treatment we added water with a high humic content. We found that carp reduced activity in response to predator cues, but only in the turbidity treatments (clay, algae), whereas the response in the brown water treatment was intermediate, and not significantly different from, clear and turbid water treatments. The increased reliance on chemical cues indicates that crucian carp can compensate for the reduced information content from vision in waters where optical conditions are degraded. The lower effect in brown water may be due to the reduction in light intensity, changes in the spectral composition (reduction of UV light) or to a change in chemical properties of the cue in humic waters.     

Hormonally derived sex pheromones in fish: exogenous cues and signals from gonad to brain

Living in a medium that can limit visual information but readily exposes the olfactory organ to hormonal compounds released by conspecifics, fish throughout their long evolutionary history have had both clear cause and ample opportunity to evolve olfactory responsiveness to these potentially important chemical cues (hormonal pheromones). Indeed, water-borne steroids, prostaglandins, and their metabolites are detected with great sensitivity and specificity by the olfactory organs of diverse fishes, and exert important effects on reproductive behavior and physiology in major taxa including carps (goldfish), catfishes, salmon, and gobies. Best understood are goldfish, where periovulatory females sequentially release a preovulatory steroid pheromone and a postovulatory prostaglandin pheromone that dramatically affect male behavior, physiology, and reproductive fitness. Although the diverse array of hormonal products released and detected by fish indicates clear potential for species-specific hormonal pheromones, olfactory recordings showing similar patterns of hormone detection among closely related species provide little evidence of selection for specificity. By demonstrating that the actions of sex hormones and related products are not limited to reproductive synchrony within the individual, the relatively recent discovery of hormonal pheromones has considerably expanded our understanding of fish reproductive function, while providing valuable model systems for future study of olfactory function and pheromone evolution.     

The trade-off between feeding, mate seeking and predator avoidance in copepods: behavioural responses to chemical cues

The behavioural responses of adult Temora longicornis to chemicalsignals from predators and potential sexual partners was studiedusing an automated three-dimensional (3-D) tracking system anda 2-D high-resolution filming set-up. The animals adapted theirswimming speed and turning behaviour in reaction to chemicalsignals to increase or decrease encounter probability with otherorganisms. Female copepods were shown to react to the smellof males with little ‘hops’, quite distinct fromboth their normal smooth swimming motion and escape responses.These hops are likely to be signals intended to increase theencounter probability with males. Swimming behaviour was shownto be a trade-off between conflicting motivations such as predatoravoidance on the one hand, and the search for food and sexualpartners on the other.     

Two threats at once: encounters with predator cues alter host life-history and morphological responses to parasite spores

Parasites and predators are ubiquitous threats in every ecosystem. Host and prey species, respectively, have evolved effective protective mechanisms which are assumed to involve costs. In this study, we analyzed potential interactions between both threats. We exposed waterfleas (Daphnia longicephala) simultaneously to parasite spores (the yeast Metschnikowia) and cues from predatory notonectids (Notonecta glauca). In response to the parasite, D. longicephala had a delayed maturation time and produced less and smaller offspring, even though the parasite developed no spores. This suggests that hosts can successfully fight off the parasite invoking defensive costs. Some of these effects were altered or even reversed by the presence of predator cues. For example, time to maturity was further delayed when the Daphnia were exposed to both threats than under parasite stress alone. In addition, more offspring were produced in the presence of both threats, although parasites alone reduced their number. However, there was no effect of parasite exposure on the expression of morphological defenses. Our results imply that the impact of parasites on host species depends strongly on the presence of further threats. Similar types of experimental approaches may enhance our understanding of the effects of multiple stressors in natural systems.     

Behavioural responses of fishes to chemical cues: results from a new bioassay

A new behavioural bioassay for determining the locomotor response by fish to chemical cues is described. The assay combines a behavioural trough, capable of delivering up to 10 discrete plumes of treated water, together with a digital method for quantifying the movements of single fish. Sample responses of two whitefish, Coregonus clupeaformis , one to an attractant (a food extract) and one to a repellant (quinine monohydrochloride), are provided. Their locomotor responses are analyzed in terms of such parameters as total distance travelled, average velocity, number of turns, average excursion distance, and the number of times specific boundaries are crossed. The assay's sensitivity is demonstrated through concentration-response curves by Arctic char, Salvelinus alpinus , to the bile acid, taurocholic acid, and the amino acids, L-serine and L-alanine. Behavioural response threshold concentrations for these three substances were estimated to be 10⁻¹⁵, 10⁻ⁿ, and 10⁻⁹ m, respectively.     

Smell, learn and live: The role of chemical alarm cues in predator learning during early life history in a marine fish

The speed with which individuals can learn to identify and react appropriately to predation threats when transitioning to new life history stages and habitats will influence their survival. This study investigated the role of chemical alarm cues in both anti-predator responses and predator identification during a transitional period in a newly settled coral reef damselfish, Pomacentrus amboinensis. Individuals were tested for changes in seven behavioural traits in response to conspecific and heterospecific skin extracts. Additionally, we tested whether fish could learn to associate a previously novel chemical cue (i.e. simulated predator scent) with danger, after previously being exposed to a paired cue combining the conspecific skin extract with the novel scent. Fish exposed to conspecific skin extracts were found to significantly decreased their feeding rate whilst those exposed to heterospecific and control cues showed no change. Individuals were also able to associate a previously novel scent with danger after only a single previous exposure to the paired conspecific skin extract/novel scent cue. Our results indicate that chemical alarm cues play a large role in both threat detection and learned predator recognition during the early post-settlement period in coral reef fishes.     

A test of the risk allocation hypothesis: tadpole responses to temporal change in predation risk

The risk allocation hypothesis predicts that temporal variationin predation risk can influence how animals allocate feedingbehavior among situations that differ in danger. We testedthe risk allocation model with tadpoles of the frog Rana lessonae,which satisfy the main assumptions of this model because theymust feed to reach metamorphosis within a single season, theirbehavioral defense against predators is costly, and they canrespond to changes in risk integrated over time. Our experiment switched tadpoles between artificial ponds with different numbersof caged dragonfly larvae and held them at high and low riskfor different portions of their lives. Tadpoles responded stronglyto predators, but they did not obey the risk allocation hypothesis:as the high-risk environment became more dangerous, there wasno tendency for tadpoles to allocate more feeding to the low-riskenvironment, and as tadpoles spent more time at risk, they didnot increase feeding in both environments. Our results suggestthat the model might be more applicable when the time spentunder high predation risk is large relative to the time requiredto collect resources.     

Antipredator behaviour mediated by chemical cues: the role of conspecific alarm signalling and predator labelling in the avoidance response of a marine gastropod

Gastropods represent a challenge in the understanding of alarm signalling. We studied predator avoidance (climbing behaviour) of the marine snail Tegula funebralis in laboratory experiments. Snails were exposed to crude extract of conspecifics, and to water conditioned by actively feeding or non-feeding predatory crabs. Crabs had previously been maintained on different diets, and were accordingly labelled by chemical cues of various origins. Tegula -extract alone released climbing behaviour in May, but not in June. However, during both these months, snails responded to chemical cues from crabs that were actively feeding on Tegula . Crabs labelled by Tegula -diet, and actively feeding on Tegula , also caused more climbing responses compared to crabs labelled by other diets. Chemical cues derived from crabs actively feeding on another snail species, or from non-feeding crabs, did not induce snail climbing no matter the previous feeding history of the predators. When snails received Tegula -extract combined with water conditioned with a non-feeding, Tegula -labelled crab, no climbing occurred. However, when the non-feeding, Tegula -labelled crab was present in the solution of Tegula -extract, moderate climbing responses were obtained. The results imply that climbing responses of T. funebralis are in general caused by the action of a two-component system. This system seems to be a mixture of chemical cues leaking from the tissue of conspecifics when being eaten, and latent conspecific chemicals that are modified in crabs and presumably released with the urine of chemically labelled predators. The modified chemical labels appear to be fully released by crabs when feeding, and moderately released when detecting food. The responses obtained in May with crude extract alone may result from a seasonal change in alarm signalling properties, or a change in behavioural responsiveness of snails exposed to a variable predator regime.     

Photoperiodic regulation of behavioral responses to bacterial and viral mimetics: a test of the winter immunoenhancement hypothesis

Siberian hamsters (Phodopus sungorus) exhibit changes in immune function following adaptation to short photoperiods, including a marked attenuation of energetically expensive thermoregulatory and behavioral responses to gram-negative bacterial infections. Whether this seasonal attenuation of the immune response is idiosyncratic to gram-negative infections or is representative of innate immune responses in general is not known. If seasonal attenuation of responsiveness to infection is indeed driven primarily by anticipation of energetic constraints, then one would predict that responsiveness to all pathogens would be diminished during short days. If, on the other hand, seasonal changes in responsiveness to infection reflect anticipation of specific pathogens that are common at different phases of the annual cycle, then one would expect short photoperiods to attenuate responsiveness to some pathogens and long photoperiods to attenuate responsiveness to others. To resolve this issue, we exposed male Siberian hamsters to either long or short photoperiods for 11 weeks, then examined their behavioral sickness responses to compounds that represent the minimally immunogenic components of gram-negative bacterial (lipopolysaccharide), gram-positive bacterial (muramyl dipeptide), and viral (polyinosinepolycytidylic acid) organisms. Hamsters exhibited anorexic, anhedonic, ponderal, and/or thermoregulatory sickness behaviors to all 3 pathogen mimetics, but in all cases in which sickness responses were evident, they were attenuated in short days. Energetically costly behavioral responses to several distinct classes of infectious organisms are attenuated in anticipation of winter. The data are not consistent with a pathogen-specific seasonal modulation of innate immune responses.     

Olfactory responses ofTrichogramma maidis Pint,et Voeg.: Effects of chemical cues and behavioral plasticity

Oriented responses of Trichogramma maidisPint, et Voeg. to airborne odors were observed in a four-armed olfactometer. Experiments were carried out with odors of host eggs, the sex pheromone of Ostrinia nubilalisHbn, and maize extract, offered singly or in combination, both to naive wasps and to wasps previously exposed to the tested odor during an oviposition experience. The exploratory behavior in the olfactometer was quantified by means of a computer program which performed a space-time analysis of the insect 's movements. Whereas the naive wasps did not respond to the odor of the eggs, the synthetic sex pheromone of O. nubilalis,or the maize extract presented singly, they did react to a mixture of these three volatile cues. Prior oviposition in the odor of maize extract or in the combination of odors induced an increased preference toward the conditioning scent. This phenomenon did not occur when the wasps were conditioned to egg odor or sex pheromone alone. These results suggest that females can learn to associate some olfactory cues with the presence of the host. Immediately following the presentation of the combination of odors, a strong attraction of experienced wasps occurred; it decreased as the experiment progressed and finally reached the level presented by naive insects. Adult conditioning to the combination of odors also resulted in reduced variability in the behavioral responses.     

Solution structure of BSTI: a new trypsin inhibitor from skin secretions of Bombina bombina

The three-dimensional solution structure of BSTI, a trypsin inhibitor from the European frog Bombina bombina, has been solved using (1)H NMR spectroscopy. The 60 amino acid protein contains five disulfide bonds, which were unambiguously determined to be Cys (4--38), Cys (13--34), Cys (17--30), Cys (21--60), and Cys (40--54) by experimental restraints and subsequent structure calculations. The main elements of secondary structure are four beta-strands, arranged as two small antiparallel beta-sheets. The overall fold of BSTI is disk shaped and is characterized by the lack of a hydrophobic core. The presumed active site is located on a loop comprising residues 21--34, which is a relatively disordered region similar to that seen in many other protease inhibitors. However, the overall fold is different to other known protease inhibitors with the exception of a small family of inhibitors isolated from nematodes of the family Ascaris and recently also from the haemolymph of Apis mellifera. BSTI may thus be classified as a new member of this recently discovered family of protease inhibitors.     

Diurnal test periods influence behavioral responses of Aedes aegypti (Diptera: Culicidae) to repellents

There is a broad understanding of the influence of environmental factors on various aspects of normal mosquito behavior. How these external factors influence responses to repellent compounds is far less clear. The objective of this study was to investigate the effect of different daytime periods combining the normal circadian activity of a laboratory colony of Aedes aegypti (L.) with behavioral responses of mosquitoes exposed to three different compounds possessing repellent properties. Using an excito-repellency test chamber with different test designs (contact irritancy + repellency and noncontact repellency), female mosquitoes were exposed to each chemical or matching blank control during four different 3-h time intervals beginning 0600 to 1800?h. Mosquitoes showed more significant avoidance responses (escape movement away from the chemical) when exposed to either DEET or hairy basil during the afternoon periods. With deltamethrin, there was no significant difference in repellent escape movement during any period of testing. Escape activity with deltamethrin was significantly greater during all diurnal periods in contact tests compared to DEET and hairy basil. From this study, it was shown that time of diurnal testing can significantly influence behavioral responses of Ae. aegypti exposed to chemical-based repellents. Therefore, the assessment of chemicals (toxins, repellents, attractants) and must carefully consider time-of-test as a potential confounding factor during evaluation and comparisons.     

Effects of androgens on behavioral and vomeronasal responses to chemosensory cues in male terrestrial salamanders (Plethodon shermani)

Chemosensory stimuli and sex steroid hormones are both required for the full expression of social behaviors in many species. The terrestrial salamander, Plethodon shermani, is an emerging nonmammalian system for investigating the nature and evolution of pheromonal communication, yet little is known regarding the role of sex steroid hormones. We hypothesized that increased circulating androgen levels in male P. shermani enhance chemoreception through morphological, behavioral, and physiological mechanisms. Experimental elevation of plasma androgens increased development of cirri, morphological structures thought to enhance the transfer of chemosensory cues from the substrate to the vomeronasal organ (VNO). Elevated plasma androgens also increased expression of a chemo-investigatory behavior (nose tapping) and increased preference for some female-derived chemosensory cues. Male-produced courtship pheromones activated a large number of cells in the VNO as measured by the method of agmatine uptake. However, androgen levels did not affect the total number of vomeronasal cells activated by male-produced courtship pheromones. Future studies will determine whether androgens potentially modulate responsiveness of the VNO to female-derived (as opposed to male-derived) chemosensory cues.     

To swim or not to swim: regional effects of serotonin, octopamine and amine mixtures in the medicinal leech

Focally treating the head brain of the medicinal leech Hirudo medicinalis with various biogenic amines affected the initiation, termination and maintenance of fictive swimming (i.e., the neural correlate of swimming). Application of serotonin to saline surrounding only the head brain inhibited fictive swimming, whereas removing serotonin induced swimming. This contrasts sharply with previous observations that serotonin applied to the nerve cord induces swimming. Although application of octopamine to the brain activated swimming, a mixture of octopamine and serotonin inhibited swimming. Subsequent removal of this mixture from the brain activated robust swimming and was more potent for activating swimming than either the removal of serotonin or the application of octopamine. Swim episodes induced by brain-specific manipulations of octopamine had more swim bursts per episode than those induced by serotonin. These brain-specific effects of the amines on fictive swimming are probably due to the modulation of higher-order circuits that control locomotion in the leech. We observed that serotonin or a mixture of serotonin and octopamine hyperpolarized an identified descending brain interneuron known as Tr2. Removal of the mixture caused Tr2 to exhibit membrane potential depolarizations that correlated in time with the expression of swim episodes.     

Embryonic stem cells and skin: from cellular model to therapeutic potential

Embryonic stem (ES) cells are pluripotent cells able to differentiate into many cell types in vitro, thus providing a potential unlimited supply of cells for cognitive in vitro studies and cell-based therapy. We recently reported their efficient ability to recapitulate ectodermal and epidermal fates and form, in culture, a multilayered epidermis coupled with an underlying dermal compartment, similar to native skin. Thus, ES cells have the potential to recapitulate the reciprocal instructive ectodermal-mesodermal commitments, characteristic of embryonic skin formation. We clarified the function of BMP-4 in the binary neuroectodermal choice by stimulating sox-1+ neural precursors to undergo specific apoptosis while inducing epidermal differentiation. We further demonstrated that p63 stimulates ectodermal cell proliferation and is necessary for epidermal commitment. We provided further evidence that this unique cellular model provides a powerful tool to identify the molecular mechanisms controlling normal skin development and to investigate human ectodermal dysplasia congenital pathologies linked to p63 (in p63-ectodermal dysplasia human congenital pathologies). Epidermal stem cell activity has been used for years to repair skin injuries, but ex vivo keratinocyte amplification has limitations and grafted skin homeostasis is not totally satisfactory. Human ES cells raise hopes that the understanding of developmental steps leading to the generation of epidermal stem cells will once be translated into therapeutic benefit. We recently demonstrated that human embryonic stem cells can give rise to a stable somatic ectodermal cell population. Its finite population doubling, normal cell cycle kinetics and the absence of teratoma formation strongly suggest that, although derived from human embryonic stem cells, these ectodermal cells represent a clinically safe somatic cell population. They could thus be particularly useful as a source for committed, homogeneous, non-tumorigenic cell populations to be employed in clinical trials for epithelial stem cell loss.