Experimental evidence that chronic outgroup conflict reduces reproductive success in a cooperatively breeding fish.

Conflicts with conspecific outsiders are common in group-living species, from ants to primates, and are argued to be an important selective force in social evolution. However, whilst an extensive empirical literature exists on the behaviour exhibited during and immediately after interactions with rivals, only very few observational studies have considered the cumulative fitness consequences of outgroup conflict. Using a cooperatively breeding fish, the daffodil cichlid (Neolamprologus pulcher), we conducted the first experimental test of the effects of chronic outgroup conflict on reproductive investment and output. 'Intruded' groups received long-term simulated territorial intrusions by neighbours that generated consistent group-defence behaviour; matched 'Control' groups (each the same size and with the same neighbours as an Intruded group) received no intrusions in the same period. Intruded groups had longer inter-clutch intervals and produced eggs with increasingly less protein than Control groups. Despite the lower egg investment, Intruded groups provided more parental care and achieved similar hatching success to Control groups. Ultimately, however, Intruded groups had fewer and smaller surviving offspring than Control groups at 1-month post-hatching. We therefore provide experimental evidence that outgroup conflict can decrease fitness via cumulative effects on reproductive success, confirming the selective potential of this empirically neglected aspect of sociality.

Fitness consequences of outgroup conflict.

In social species across the animal kingdom, conspecific outsiders threaten the valuable resources of groups and their members. This outgroup conflict is recognised as a powerful selection pressure, but we argue that studies explicitly quantifying the fitness consequences need to be broader in scope: more attention should be paid to delayed, cumulative, and third-party fitness consequences, not just those arising immediately to group members involved in physical contests. In the first part of this review, we begin by documenting how single contests can have survival and reproductive consequences either immediately or with a delay. Then, we step beyond contests to describe fitness consequences that can also result from interactions with cues of rival presence and the general landscape of outgroup threat, and beyond single interactions to describe cumulative effects of territorial pressure and elevated outgroup-induced stress. Using examples from a range of taxa, we discuss which individuals are affected negatively and positively, considering both interaction participants and third-party group members of the same or the next generation. In the second part of the review, we provide suggestions about how to move forward. We highlight the importance of considering how different types of outgroup conflict can generate different selection pressures and of investigating variation in fitness consequences within and between species. We finish by discussing the value of theoretical modelling and long-term studies of natural populations, experimental manipulations, and meta-analyses to develop further our understanding of this crucial aspect of sociality.

Variation between species, populations, groups and individuals in the fitness consequences of out-group conflict.

Out-group conflict is rife in the natural world, occurring from primates to ants. Traditionally, research on this aspect of sociality has focused on the interactions between groups and their conspecific rivals, investigating contest function and characteristics, which group members participate and what determines who wins. In recent years, however, there has been increasing interest in the consequences of out-group conflict. In this review, we first set the scene by outlining the fitness consequences that can arise immediately to contest participants, as well as a broader range of delayed, cumulative and third-party effects of out-group conflict on survival and reproductive success. For the majority of the review, we then focus on variation in these fitness consequences of out-group conflict, describing known examples both between species and between populations, groups and individuals of the same species. Throughout, we suggest possible reasons for the variation, provide examples from a diverse array of taxa, and suggest what is needed to advance this burgeoning area of social evolution. This article is part of the theme issue 'Intergroup conflict across taxa'.

Impacts of additional noise on the social interactions of a cooperatively breeding fish.

Anthropogenic noise is a global pollutant known to affect the behaviour of individual animals in all taxa studied. However, there has been relatively little experimental testing of the effects of additional noise on social interactions between conspecifics, despite these forming a crucial aspect of daily life for most species. Here, we use established paradigms to investigate how white-noise playback affects both group defensive actions against an intruder and associated within-group behaviours in a model fish species, the cooperatively breeding cichlid Neolamprologus pulcher. Additional noise did not alter defensive behaviour, but did result in changes to within-group behaviour. Both dominant and subordinate females, but not the dominant male, exhibited less affiliation and showed a tendency to produce more submissive displays to groupmates when there was additional noise compared with control conditions. Thus, our experimental results indicate the potential for anthropogenic noise to affect social interactions between conspecifics and emphasize the possibility of intraspecific variation in the impacts of this global pollutant.

Pipefish embryo oxygenation, survival, and development: egg size, male size, and temperature effects.

In animals with uniparental care, the quality of care provided by one sex can deeply impact the reproductive success of both sexes. Studying variation in parental care quality within a species and which factors may affect it can, therefore, shed important light on patterns of mate choice and other reproductive decisions observed in nature. Using Syngnathus typhle, a pipefish species with extensive uniparental male care, with embryos developing inside a brood pouch during a lengthy pregnancy, we assessed how egg size (which correlates positively with female size), male size, and water temperature affect brooding traits that relate to male care quality, all measured on day 18, approximately 1/3, of the brooding period. We found that larger males brooded eggs at lower densities, and their embryos were heavier than those of small males independent of initial egg size. However, large males had lower embryo survival relative to small males. We found no effect of egg size or of paternal size on within-pouch oxygen levels, but oxygen levels were significantly higher in the bottom than the middle section of the pouch. Males that brooded at higher temperatures had lower pouch oxygen levels presumably because of higher embryo developmental rates, as more developed embryos consume more oxygen. Together, our results suggest that small and large males follow distinct paternal strategies: large males positively affect embryo size whereas small males favor embryo survival. As females prefer large mates, offspring size at independence may be more important to female fitness than offspring survival during development.

Experimental evidence that intruder and group member attributes affect outgroup defence and associated within-group interactions in a social fish.

In many social species, individuals communally defend resources from conspecific outsiders. Participation in defence and in associated within-group interactions, both during and after contests with outgroup rivals, is expected to vary between group members because the threat presented by different outsiders is not the same to each individual. However, experimental tests examining both the contributions to, and the consequences of, outgroup conflict for all group members are lacking. Using groups of the cichlid Neolamprologus pulcher, we simulated territorial intrusions by different-sized female rivals and altered the potential contribution of subordinate females to defence. Dominant females and subordinate females defended significantly more against size- and rank-matched intruders, while males displayed lower and less variable levels of defence. Large and small, but not intermediate-sized, intruders induced increased levels of within-group aggression during intrusions, which was targeted at the subordinate females. Preventing subordinate females from helping in territorial defence led to significant decreases in post-contest within-group and female-specific submissive and affiliative displays. Together, these results show that the defensive contributions of group members vary greatly depending both on their own traits and on intruder identity, and this variation has significant consequences for within-group social dynamics both during and in the aftermath of outgroup contests.

Meerkat close calling patterns are linked to sex, social category, season and wind, but not fecal glucocorticoid metabolite concentrations.

It is well established that animal vocalizations can encode information regarding a sender's identity, sex, age, body size, social rank and group membership. However, the association between physiological parameters, particularly stress hormone levels, and vocal behavior is still not well understood. The cooperatively breeding African meerkats (Suricata suricatta) live in family groups with despotic social hierarchies. During foraging, individuals emit close calls that help maintain group cohesion. These contact calls are acoustically distinctive and variable in rate across individuals, yet, information on which factors influence close calling behavior is missing. The aim of this study was to identify proximate factors that influence variation in call rate and acoustic structure of meerkat close calls. Specifically, we investigated whether close calling behavior is associated with sex, age and rank, or stress hormone output (i.e., measured as fecal glucocorticoid metabolite (fGCM) concentrations) as individual traits of the caller, as well as with environmental conditions (weather) and reproductive seasonality. To disentangle the effects of these factors on vocal behavior, we analyzed sound recordings and assessed fGCM concentrations in 64 wild but habituated meerkats from 9 groups during the reproductive and non-reproductive seasons. Dominant females and one-year old males called at significantly higher rates compared to other social categories during the reproductive season. Additionally, dominant females produced close calls with the lowest mean fundamental frequencies (F0) and the longest mean pulse durations. Windy conditions were associated with significantly higher call rates during the non-reproductive season. Fecal GCM concentrations were unrelated to close calling behavior. Our findings suggest that meerkat close calling behavior conveys information regarding the sex and social category of the caller, but shows no association with fGCM concentrations. The change in call rate in response to variation in the social and ecological environments individuals experience indicates some degree of flexibility in vocal production.

Exorcising Grice's ghost: an empirical approach to studying intentional communication in animals.

Language's intentional nature has been highlighted as a crucial feature distinguishing it from other communication systems. Specifically, language is often thought to depend on highly structured intentional action and mutual mindreading by a communicator and recipient. Whilst similar abilities in animals can shed light on the evolution of intentionality, they remain challenging to detect unambiguously. We revisit animal intentional communication and suggest that progress in identifying analogous capacities has been complicated by (i) the assumption that intentional (that is, voluntary) production of communicative acts requires mental-state attribution, and (ii) variation in approaches investigating communication across sensory modalities. To move forward, we argue that a framework fusing research across modalities and species is required. We structure intentional communication into a series of requirements, each of which can be operationalised, investigated empirically, and must be met for purposive, intentionally communicative acts to be demonstrated. Our unified approach helps elucidate the distribution of animal intentional communication and subsequently serves to clarify what is meant by attributions of intentional communication in animals and humans.

Evolutionary ecology of pipefish brooding structures: embryo survival and growth do not improve with a pouch.

For animals that reproduce in water, many adaptations in life-history traits such as egg size, parental care, and behaviors that relate to embryo oxygenation are still poorly understood. In pipefishes, seahorses and seadragons, males care for the embryos either in some sort of brood pouch, or attached ventrally to the skin on their belly or tail. Typically, egg size is larger in the brood pouch group and it has been suggested that oxygen supplied via the pouch buffers the developing embryos against hypoxia and as such is an adaptation that has facilitated the evolution of larger eggs. Here, using four pipefish species, we tested whether the presence or absence of brood pouch relates to how male behavior, embryo size, and survival are affected by hypoxia, with normoxia as control. Two of our studied species Entelurus aequoreus and Nerophis ophidion (both having small eggs) have simple ventral attachment of eggs onto the male trunk, and the other two, Syngnathus typhle (large eggs) and S. rostellatus (small eggs), have fully enclosed brood pouches on the tail. Under hypoxia, all species showed lower embryo survival, while species with brood pouches suffered greater embryo mortality compared to pouchless species, irrespective of oxygen treatment. Behaviorally, species without pouches spent more time closer to the surface, possibly to improve oxygenation. Overall, we found no significant benefits of brood pouches in terms of embryo survival and size under hypoxia. Instead, our results suggest negative effects of large egg size, despite the protection of brood pouches.

Validation of a Fecal Glucocorticoid Assay to Assess Adrenocortical Activity in Meerkats Using Physiological and Biological Stimuli.

In mammals, glucocorticoid (i.e. GC) levels have been associated with specific life-history stages and transitions, reproductive strategies, and a plethora of behaviors. Assessment of adrenocortical activity via measurement of glucocorticoid metabolites in feces (FGCM) has greatly facilitated data collection from wild animals, due to its non-invasive nature, and thus has become an established tool in behavioral ecology and conservation biology. The aim of our study was to validate a fecal glucocorticoid assay for assessing adrenocortical activity in meerkats (Suricata suricatta), by comparing the suitability of three GC enzyme immunoassays (corticosterone, 11ß-hydroxyetiocholanolone and 11oxo-etiocholanolone) in detecting FGCM increases in adult males and females following a pharmacological challenge with adrenocorticotropic hormone (ACTH) and biological stimuli. In addition, we investigated the time course characterizing FGCM excretion, the effect of age, sex and time of day on FGCM levels and assessed the potential effects of soil contamination (sand) on FGCM patterns. Our results show that the group specific 11ß-hydroxyetiocholanolone assay was most sensitive to FGCM alterations, detecting significant and most distinctive elevations in FGCM levels around 25 h after ACTH administration. We found no age and sex differences in basal FGCM or on peak response levels to ACTH, but a marked diurnal pattern, with FGCM levels being substantially higher in the morning than later during the day. Soil contamination did not significantly affect FGCM patterns. Our results emphasize the importance of conducting assay validations to characterize species-specific endocrine excretion patterns, a crucial step to all animal endocrinology studies using a non-invasive approach.

The evolutionary puzzle of egg size, oxygenation and parental care in aquatic environments.

Offspring fitness generally improves with increasing egg size. Yet, eggs of most aquatic organisms are small. A common but largely untested assumption is that larger embryos require more oxygen than they can acquire through diffusion via the egg surface, constraining egg size evolution. However, we found no detrimental effects of large egg size on embryo growth and survival under hypoxic conditions. We tested this in the broad-nosed pipefish, Syngnathus typhle, whose males provide extensive care (nourishment, osmoregulation and oxygenation) to their young in a brood pouch on their bodies. We took advantage of this species' pronounced variation in egg size, correlating positively with female size, and tested the effect of hypoxia (40% dissolved oxygen) versus fully oxygenated (100%) water on embryo size and survival of large versus small eggs after 18 days of paternal brooding. Egg size did not affect embryo survival, regardless of O2 treatment. While hypoxia affected embryo size negatively, both large and small eggs showed similar reductions in growth. Males in hypoxia ventilated more and males with large eggs swam more, but neither treatment affected their position in the water column. Overall, our results call into question the most common explanation for constrained egg size evolution in aquatic environments.