The modulation of social and non-social behavior by arginine vasotocin in the common waxbill, Estrilda astrild.

The influence of the neuropeptide arginine vasotocin (AVT) has been demonstrated across various species, on an ample range of behaviors, yet the results appear to be highly species-specific. In this study, we aimed to test how AVT influences both social and non-social behaviors in the common waxbill Estrilda astrild, a highly social estrildid finch. Through a within-subject design study, we experimentally manipulated AVT pathways through muscular injections of both an agonist and an antagonist of AVT at different dosages, and performed competition over food tests to assess behavioral changes. Our observations reveal a decrease in birds' movements with both low and high dosages of AVT. Additionally, the higher AVT dosage led to a significant decrease in birds' feeding, aggressive behavior, and allopreening. Conversely, the lower AVT dosage increased the duration of allopreening, which is a proxy for affiliation. The use of Manning Compound, a V1a antagonist, did not produce any changes in behavior, however, the absence of affinity studies for this compound in birds makes it difficult to interpret these results. It is plausible that in common waxbills, AVT V1b or V1a receptors may be involved in regulating movement, feeding, aggressive behavior, and allopreening, rather than V2 AVT receptors.

Learning competence of the temperate facultative cleaner wrasse Thalassoma pavo (Labridae).

Cleaner fishes remove parasites from other fishes called "clients," thereby contributing to a healthier ecosystem. Although the behavior and learning abilities of dedicated and tropical cleaner fishes have been broadly studied, a limited number of studies investigated the behavior of facultative and temperate cleaner fishes and, to the best of our knowledge, none focused on their cognitive abilities. Here, we tested the learning abilities of a species of temperate facultative cleaner, the ornate wrasse Thalassoma pavo in laboratory conditions, based on two problems. These two problems, or tasks, are relevant in two different contexts: the first one, deemed as mutualistically relevant, the cue-based discrimination task, and the second one, the spatial-based discrimination task, which is relevant in a non-cleaning context, when fish navigate through their environment to find food and return to their territories. We found that T. pavo individuals were able to solve these two tasks but excelled at the spatial task rather than with the cue discrimination. The same individuals were also challenged to learn the reverse protocol of these tasks and were again most successful in learning the reverse spatial discrimination problem, but not the cue. Contrary to the dedicated cleaner wrasse Labroides dimidiatus, this temperate facultative cleaner wrasse is slower to learn mutualistic relevant problems but competent in solving spatially derived problems. This may be due to the specific demands of their socio-ecological environment, with facultative cleaners having a greater component of non-mutualistic skills (spatial component), which prepares them to search for alternative food sources if necessary (e.g., feeding on the substrate) or even to expand territories more easily and less prepared to deal with mutualistic exchanges compared to dedicated cleaners that specialize to become increasingly socially competent.

Comparative Brain Morphology of Cleaning and Sponge-Dwelling Elacatinus Gobies.

INTRODUCTION: Comparative studies of brain anatomy between closely related species have been very useful in demonstrating selective changes in brain structure. Within-species comparisons can be particularly useful for identifying changes in brain structure caused by contrasting environmental selection pressures. Here, we aimed to understand whether differences within and between species in habitat use and foraging behaviour influence brain morphology, on both ecological and evolutionary time scales. METHODS: We used as a study model three species of the Elacatinus genus that differ in their habitat-foraging mode. The obligatory cleaning goby Elacatinus evelynae inhabits mainly corals and feeds mostly on ectoparasites removed from larger fish during cleaning interactions. In contrast, the obligatory sponge-dwelling goby Elacatinus chancei inhabits tubular sponges and feeds on microinvertebrates buried in the sponges' tissues. Finally, in the facultatively cleaning goby Elacatinus prochilos, individuals can adopt either phenotype, the cleaning or the sponge-dwelling habitat-foraging mode. By comparing the brains of the facultative goby phenotypes to the brains of the obligatory species we can test whether brain morphology is better predicted by phylogenetic relatedness or the habitat-foraging modes (cleaning × sponge dwelling). RESULTS: We found that E. prochilos brains from both types (cleaning and sponge dwelling) were highly similar to each other. Their brains were in general more similar to the brains of the most closely related species, E. evelynae (obligatory cleaning species), than to the brains of E. chancei (sponge-dwelling species). In contrast, we found significant brain structure differences between the cleaning species (E. evelynae and E. prochilos) and the sponge-dwelling species (E. chancei). These differences revealed independent changes in functionally correlated brain areas that might be ecologically adaptive. E. evelynae and E. prochilos had a relatively larger visual input processing brain axis and a relatively smaller lateral line input processing brain axis than E. chancei. CONCLUSION: The similar brain morphology of the two types of E. prochilos corroborates other studies showing that individuals of both types can be highly plastic in their social and foraging behaviours. Our results in the Elacatinus species suggest that morphological adaptations of the brain are likely to be found in specialists whereas species that are more flexible in their habitat may only show behavioural plasticity without showing anatomical differences.

First record of intraspecific cleaning behavior by the threatened Leuciscidae Squalius alburnoides (Steindachner, 1866) at the Guadiana River basin (Portugal).

Squalius alburnoides (Steindachner, 1866) is an endemic threatened species from the Iberian Peninsula. Here, we report the first observations of intraspecific cleaning behavior in isolated summer pools in the Guadiana River Basin (Portugal). We found that focal S. alburnoides solicited cleaning by adopting an immobile tail-stand position known as "posing," which immediately signaled a response to a few conspecifics that approached and inspect them. Our study expands the list of cleanerfish species in freshwaters, giving emphasis to the importance of mutual positive behavior within an endangered species, particularly when facing seasonal disturbance.

Effect of Tryptophan Dietary Content on Meagre, Argyrosomus regius, Juveniles Stress and Behavioral Response.

There are a high number of stressors present in aquaculture that can affect fish welfare and quality. One way of mitigating stress response is by increasing dietary tryptophan. In this study, three diets containing 0.5% (Tript1), 0.6% (Tript2), and 0.8% (Tript3) of tryptophan were tested in 32 g juvenile meagre for 56 days. At the end of the trial, survival, growth, and proximate composition were similar between treatments. Significant differences were found in the plasma parameters before and after a stress test consisting of 30 s of air exposure. Blood glucose levels were higher in the post-stress for all treatments (e.g., 63.9 and 76.7 mg/dL for Tript1 before and after the stress test), and the hemoglobin values were lower in the post-stress of Tript1 (1.9 g/dL compared to 3.0 and 2.4 g/dL for Tript2 and Tript3, respectively). In terms of behavior, three tests were carried out (novel tank diving and shoaling assays, and lateralization test), but no significant differences were found, except for the number of freezing episodes during the anxiety test (1.4 for Tript3 compared to 3.5 and 4.2 for the other treatments). This study suggests that supplementation with dietary tryptophan, particularly in higher dosage (0.8%), can reduce anxiety-like behavior in meagre exposure to acute stress (novel tank). Although the remaining results showed mild effects, they provide some clues as to the potential of this amino acid as a stress mitigator in aquaculture.

Effects of mesotocin manipulation on the behavior of male and female common waxbills.

The oxytocin family of neuropeptides is implicated in the regulation of sociality across vertebrates. Non-mammalian homologs of oxytocin, such as isotocin in fish and mesotocin in amphibians, reptiles and birds, all play crucial roles modulating social and reproductive behavior. In this study, we exogenously manipulated the mesotocinergic system in a highly social bird, the common waxbill Estrild astrild, and tested the effects on affiliative and aggressive behavior by performing tests of competition over food. Birds treated with mesotocin decreased almost all the behaviors we studied (movement, feeding, allopreening), while birds treated with an oxytocin antagonist showed a reduction only in social behaviors (aggressions and allopreening). We also found two sex-specific effects: mesotocin reduced allopreening more in males than females, and the oxytocin antagonist reduced aggressiveness only in females. Our results suggest sex-specific effects in the modulation of affiliative and aggressive behaviors via mesotocinergic pathways.

Reef Location and Client Diversity Influence the Skin Microbiome of the Caribbean Cleaner Goby Elacatinus evelynae.

Fish-associated microorganisms are known to be affected by the environment and other external factors, such as microbial transfer between interacting partners. One of the most iconic mutualistic interactions on coral reefs is the cleaning interactions between cleaner fishes and their clients, during which direct physical contact occurs. Here, we characterized the skin bacteria of the Caribbean cleaner sharknose goby, Elacatinus evelynae, in four coral reefs of the US Virgin Islands using sequencing of the V4 region of the 16S rRNA gene. We specifically tested the relationship between gobies' level of interaction with clients and skin microbiota diversity and composition. Our results showed differences in microbial alpha- and beta-diversity in the skin of gobies from different reef habitats and high inter-individual variation in microbiota diversity and structure. Overall, the results showed that fish-to-fish direct contact and specifically, access to a diverse clientele, influences the bacterial diversity and structure of cleaner gobies' skin. Because of their frequent contact with clients, and therefore, high potential for microbial exchange, cleaner fish may serve as models in future studies aiming to understand the role of social microbial transfer in reef fish communities.

Oestradiol reduces female bill colour in a mutually ornamented bird.

Carotenoid-based colour signals can be costly to produce and maintain, and trade-offs between signalling and other fitness traits are expected. In mutually ornamented species, trade-offs with reproduction may be stronger for females than males, because females often dedicate more resources to offspring production, which may lead to plastic investment in colour signals and plastic sexual dichromatism. Oestradiol is a candidate mediator of this trade-off because it regulates reproductive physiology and may also influence the expression of coloration. We tested this hypothesis by giving female common waxbills (Estrilda astrild) either oestradiol (17ß-oestradiol) or empty implants during the early breeding season and measured spectral reflectance of carotenoid-based bill coloration weekly for two months. Using a model of avian vision, we found that bill colour in oestradiol-implanted females became less saturated, less red in hue and brighter, compared with control females and with unimplanted males. This resulted in a change in bill sexual dichromatism from imperceptible to perceptible. Results support the hypothesis that female reproductive physiology influences investment in coloration through changes in oestradiol and show a form of female-driven plastic sexual dichromatism. Greater sensitivity of female colour to physiological and/or environmental conditions helps explain why differences in sexual dichromatism among species differing in ecology often evolve owing to changes in female rather than male phenotype.

Hemolymph Glycemia as an Environmental Stress Biomarker in the Invasive Red Swamp Crayfish (Procambarus clarkii).

AbstractSeveral freshwater crayfish species, including Procambarus clarkii, are both ecologically important and commercially important benthic macroinvertebrates, remarkable for their potential to adapt and reproduce but also for their unique abilities to face distinct abiotic and biotic environments and become successful invaders. While much work has been done to study crayfish introductions, less focus has been given to how crayfish cope with pollution and other environmental stressors, in terms of physiological responses, and whether crayfish responses can be used to assess the effective state of their living environment. Here, we used a mixed approach combining laboratory experiments with field data to validate the use of hemolymph glucose as a relevant biomarker of red swamp crayfish (Procambarus clarkii) stress response. Three meaningful sampling locations were chosen across southern Portugal that are representative of different environments where crayfish live and are frequently captured for human consumption but also correspond to different pollution levels. To reference field measurements of glucose levels, we performed two lab-based experiments: (a) crayfish were exposed to different levels of stress (stress challenge) and (b) crawfish were exposed to a maze dispersal test, with or without water. Crayfish glucose levels were responsive to induced stress but were not correlated with dispersal efforts. Wild crayfish's body condition and stress levels responded differently to environmental conditions, with more challenged individuals showing higher glycemia levels but similar body condition. The glucose levels of the more stressed wild crayfish were visually similar to lab-based crayfish subjected to the higher stress levels (electric shocks), while the levels of glucose of crayfish at the less polluted site corresponded to those measured before the start of the challenge (baseline). The maintenance of high levels of glycemia in crayfish inhabiting more challenging habitats is revealing of their higher energetic demand state. Since P. clarkii ia globally distributed and easily sampled invasive species, quantifying its hemolymph glucose levels can be a particularly useful proxy for assessing environmental quality.

Dopamine modulates social behaviour in cooperatively breeding fish.

Dopamine is part of the reward system triggering the social decision-making network in the brain. It has hence great potential importance in the regulation of social behaviour, but its significance in the control of behaviour in highly social animals is currently limited. We studied the role of the dopaminergic system in social decision-making in the cooperatively breeding cichlid fish, Neolamprologus pulcher, by blocking or stimulating the dopaminergic D1-like and D2-like receptors. We first tested the effects of different dosages and timing of administration on subordinate group members' social behaviour within the group in an unchallenging environment. In a second experiment we pharmacologically manipulated D1-like and D2-like receptors while experimentally challenging N. pulcher groups by presenting an egg predator, and by increasing the need for territory maintenance through digging out sand from the shelter. Our results show that the D1-like and D2-like receptor pathways are differently involved in the modulation of aggressive, submissive and affiliative behaviours. Interestingly, the environmental context seems particularly crucial regarding the role of the D2-like receptors in behavioural regulation of social encounters among group members, indicating a potential pathway in agonistic and cooperative interactions in a pay-to-stay scenario. We discuss the importance of environmental information in mediating the role of dopamine for the modulation of social behaviour.

A test of context- and sex-dependent dopaminergic effects on the behavior of a gregarious bird, the common waxbill, Estrilda astrild.

The dopaminergic (DAergic) system has well-known influences on behavioral and cognitive functions. Previous work with common waxbills (Estrilda astrild) reported context-specific DAergic effects that could have been due to social environment. Manipulating the dopamine D2-like receptor family (D2R) pathways had opposing effects on behavior depending on whether waxbills were tested alone or in a small cage with a mirror as a social stimulus. As waxbills are highly gregarious, it was hypothesized that being alone or perceiving that they have a companion might explain this context dependence. To test context-dependent DAergic effects, we compared behavioral effects of D2R manipulation in waxbills in the same familiar environment, but either alone or with a familiar, same-sex companion. We found that D2R agonism decreased movement and feeding, similar to previous results when testing waxbills alone. However, contrary to the hypothesis of dependence on social context, we found that the behavioral effects of the D2R agonist were unchanged when waxbills were tested with a companion. The context dependence reported earlier might thus be due to other factors, such as the stress of being in a novel environment (small cage) or with an unfamiliar social stimulus (mirror image). In tests with a companion, we also found a sex-specific social effect of D2R manipulation: D2R blocking tended to decrease aggression in males but to increase it in females. Together with past work, our results suggest that DAergic effects on behavior involve different types of context or sex dependence.

Understanding early-life pain and its effects on adult human and animal emotionality: Translational lessons from rodent and zebrafish models.

Critical for organismal survival, pain evokes strong physiological and behavioral responses in various sentient species. Clinical and preclinical (animal) studies markedly increase our understanding of biological consequences of developmental (early-life) adversity, as well as acute and chronic pain. However, the long-term effects of early-life pain exposure on human and animal emotional responses remain poorly understood. Here, we discuss experimental models of nociception in rodents and zebrafish, and summarize mounting evidence of the role of early-life pain in shaping emotional traits later in life. We also call for further development of animal models to probe the impact of early-life pain exposure on behavioral traits, brain disorders and novel therapeutic treatments.

Pro-social and anxiolytic-like behavior following a single 24-h exposure to 17ß-estradiol in adult male zebrafish.

Estradiol (17ß-estradiol, E2) is a crucial estrogen hormone that regulates sexual, cognitive, social and affective behaviors in various species. However, complex central nervous system (CNS) effects of E2, including its activity in males, remain poorly understood. The zebrafish (Danio rerio) is rapidly becoming a powerful novel model system in translational neuroscience research. Here, we evaluate the effects of a single 24-h exposure to 20 µg/L of E2 on behavioral and endocrine (cortisol) responses in adult male zebrafish. Overall, E2 exerted pro-social effect in the social preference test, reduced whole-body cortisol levels, elevated exploration in the novel tank test and increased the shoal size in the shoaling test, indicative of an anxiolytic-like profile of this hormone in male zebrafish. Supporting mounting human and rodent evidence on the role of E2 in behavioral regulation, the observed pro-social and anxiolytic-like effects of E2 in male zebrafish reinforce the use of this aquatic organism in studying steroid-mediated CNS mechanisms of complex affective and social behaviors.

Cleaner blues: Condition-dependent colour and cleaner fish service quality.

While vivid colours in sexual signals can provide information on individual quality, vivid colours in interspecific signals have been interpreted mostly as indicating species identity and maximizing signal detection. Here we investigate if colour differences in an interspecific signal could also indicate relevant aspects of individual quality because, similarly to sexual signalling, in interspecific communication it could sometimes be advantageous to assess individual quality. For example when interacting with cleaner species, clients should benefit from assessing which individual cleaners provide better service. Since oxidative stress commonly influence condition-dependent colour signals, we oxidized the diet of cleaner wrasses (Labroides dimidiatus) in laboratory conditions to test if this affects their vivid blue skin colour and the quality of their cleaning service. Compared to controls, experimental cleaners decreased blue colour saturation after the oxidized diet treatment, decreased the quality of their cleaning service by performing less tactile stimulation and, although clients in the laboratory were de-parasitized, also tended to decrease the touching the client with the mouth. We used visual modelling, based on the spectral sensitivity of vision in various client species, to show that some client species can perceive these changes in cleaner blue colour saturation, while other client species not. We suggest that, similarly to sexual signals, some vivid colours used in interspecific communication may convey information on aspects of individual quality that are relevant to heterospecifics.

Dopamine and serotonin mediate the impact of stress on cleaner fish cooperative behavior.

Stress is known to modulate behavioral responses and rapid decision-making processes, especially under challenging contexts which often occur in social and cooperative interactions. Here, we evaluated the effects of acute stress on cooperative behavior of the Indo-Pacific cleaner wrasse (Labroides dimidiatus) and the implications of pre-treatment with monoaminergic compounds: the selective serotonin reuptake inhibitor - fluoxetine, the 5-HT1A receptor antagonist - WAY-100,635, the D1 receptor agonist - SKF-38393, and the D1 receptor antagonist - SCH-23390. We demonstrated that stress decreased the predisposal to interact and increased cortisol levels in cleaners, which are alleviated by fluoxetine and the dopaminergic D1 antagonist. Overall, our findings highlight the crucial influence of stress on cooperative behavior.

Experimental evidence for a role of dopamine in avian personality traits.

There is increasing interest in the genetic and physiological bases of behavioural differences among individuals, namely animal personality. One particular dopamine (DA) receptor gene (the dopamine receptor D4 gene) has been used as candidate gene to explain personality differences, but with mixed results. Here, we used an alternative approach, exogenously manipulating the dopaminergic system and testing for effects on personality assays in a social bird species, the common waxbill (Estrilda astrild). We treated birds with agonists and antagonists for DA receptors of both D1 and D2 receptor pathways (the latter includes the D4 receptor) and found that short-term manipulation of DA signalling had an immediate effect on personality-related behaviours. In an assay of social responses (mirror test), manipulation of D2 receptor pathways reduced time spent looking at the social stimulus (mirror image). Blocking D2 receptors reduced motor activity in this social assay, while treatment with a D2 receptor agonist augmented activity in this social assay but reduced activity in a non-social behavioural assay. Also, in the non-social assay, treatment with the D1 receptor antagonist markedly increased time spent at the feeder. These results show distinct and context-specific effects of the dopaminergic pathways on waxbill personality traits. Our results also suggest that experimental manipulation of DA signalling can disrupt a behavioural correlation (more active individuals being less attentive to mirror image) that is habitually observed as part of a behavioural syndrome in waxbills. We discuss our results in the context of animal personality, and the role of the DA system in reward and social behaviour.

Emotional behavior in aquatic organisms? Lessons from crayfish and zebrafish.

Experimental animal models are a valuable tool to study the neurobiology of emotional behavior and mechanisms underlying human affective disorders. Mounting evidence suggests that various aquatic organisms, including both vertebrate (e.g., zebrafish) and invertebrate (e.g., crayfish) species, may be relevant to study animal emotional response and its deficits. Ideally, model organisms of disease should possess considerable genetic and physiological homology to mammals, display robust behavioral and physiological responses to stress, and should be sensitive to a wide range of drugs known to modulate stress and affective behaviors. Here, we summarize recent findings in the field of zebrafish- and crayfish-based tests of stress, anxiety, aggressiveness and social preference, and discuss further perspectives of using these novel model organisms in translational biological psychiatry. Outlining the remaining questions in this field, we also emphasize the need in further development and a wider use of crayfish and zebrafish models to study the pathogenesis of affective disorders.

Testosterone causes pleiotropic effects on cleanerfish behaviour.

Mathematical modelling regarding evolutionary theory typically assumes that optimal strategies are not constrained through mechanistic processes. In contrast, recent studies on brain anatomy and neurobiology suggest that flexibility in social behaviour is rather constrained by the physiological state of the social decision-making network. Changing its state may yield selective advantages in some social contexts but neutral or even detrimental effects in others. Here we provide field evidence for such physiological trade-offs. We subjected wild female cleaner wrasse to injections of testosterone or of saline solution (control) and then observed both intraspecific interactions and interspecific cleaning behaviour with other reef fish, referred to as clients. Testosterone-treated females intensified intraspecific social interactions, showing more aggression towards smaller females and tendencies of increased aggressive and affiliative contacts with dominant males. Such testosterone-mediated changes fit the hypothesis that an increase in testosterone mediates female's focus on status in this protogynous hermaphrodite species, where females eventually change sex to become males. Moreover, we also identified other effects on interspecific social interactions: testosterone-treated females interacted less with client reef fishes and hence obtained less food. Most importantly, they selectively reduced service quality for species that were less likely to punish after being cheated. Overall, our findings suggest that testosterone causes pleiotropic effects on intra and interspecific social behaviour by broadly influencing female cleaners' decision-making.

Ecological differences in the facultative Caribbean cleaning goby Elacatinus prochilos do not predict learning performance in discriminatory two-choice tasks.

The ecological approach to comparative cognition emphasizes that the ecological and social environment are important predictors of cognitive performance. We used this approach to test whether differences in habitat use and social behavior in the facultative Caribbean cleaning goby Elacatinus prochilos predict differences in learning performance in two discriminatory two-choice tasks. This species has two behavioral ecotypes: one that frequently engages in cleaning interactions and inhabits corals in male-female pairs (cleaning gobies) and another that rarely engages in cleaning interactions and inhabits barrel sponges in large groups (sponge-dwellers). We predicted that cleaning gobies would outperform sponge-dwellers in a pattern-cued task, which consisted of identifying the pattern on a plate that consistently provided food, while sponge-dwellers would outperform cleaning gobies in a spatial task, which consisted of identifying the location of the plate. Contrary to our predictions, there was no difference in performance between the two ecotypes. Most of the gobies performed poorly in the pattern-cued task and well in the spatial task. A possible explanation for these results is that the association of a pattern with positive and negative reinforcement may not be a pre-requisite for engaging in cleaning interactions, while spatial skills might be equally required in both ecotypes. Alternatively, the two ecotypes can flexibly adjust to new feeding conditions, which would explain their similar performance in the spatial task. Further research should investigate which aspects of E. prochilos' social and ecological environment might impose challenges that require spatial cognition and whether individuals can flexibly adjust to new habitats and feeding conditions.