Maternal Care under Large Clutches with Small Eggs: The Evolution of Life History Traits in Shield Bugs.

AbstractPatterns in the correlated evolution of parental care and life history traits are long established but controversial. Although parental care is related to large egg size in many taxa, conflicting results have also been reported. To test the evolutionary relationships between parental care and life history traits, we performed phylogenetic comparative analyses using shield bugs (Heteroptera: Acanthosomatidae), in which maternal guarding of eggs and young has repeatedly evolved. Our analyses revealed that female body size affected reproductive resource allocation. Contrary to the expectations of current theories, the acquisition of maternal care was associated with small eggs, large clutches, and large egg resource allocation. There was a greater trade-off between egg size and clutch size in caring species than in noncaring species. Egg and hatchling developmental rates were not correlated with egg size but were slower in caring species than in noncaring species. Analyses of evolutionary transitions suggest that the establishment of large clutches, small eggs, and large egg resource allocation preceded the evolution of maternal care. To our knowledge, this is the first study clarifying the evolution of parental care linked with small eggs in invertebrates.

Body size evolution in otters distinguished from terrestrial mustelids.

Some taxa of mammals live in water, all of which evolved from land-dwelling ancestors. In the family Mustelidae (Mammalia: Carnivora), most species live on land, while otters, comprising the subfamily Lutrinae, inhabit aquatic environments, which include the almost exclusively aquatic sea otters (Enhydra lutris). Thus, the transition from a terrestrial to an aquatic lifestyle has occurred within this family. Despite potentially different selection pressures on body size in aquatic and terrestrial habitats, no divergence in the evolutionary pattern of body size between otters and other mustelids has previously been shown using models of trait evolution on a phylogeny. We applied models that explicitly incorporated lineage-specific directional selection to the evolution of body mass in living mustelids. Using a simulation-based likelihood and approximate Bayesian computation approach, we demonstrated lineage-specific directional selection for larger body mass in otters, which is distinct from other mustelids. There was no evidence of a difference between sea otters and other otters in the strength of directional selection for larger body mass. Additionally, our analyses supported no difference in the rate at which body mass evolves in both directions between otters and other mustelids. These findings suggest that the evolution of body mass in otters is associated with selective advantages of larger size rather than the relaxation of constraints on body size in aquatic habitats, like other aquatic mammals such as sirenians, cetaceans, and pinnipeds.

CD44 correlates with longevity and enhances basal ATF6 activity and ER stress resistance.

The naked mole rat (NMR) is the longest-lived rodent, resistant to multiple age-related diseases including neurodegeneration. However, the mechanisms underlying the NMR's resistance to neurodegenerative diseases remain elusive. Here, we isolated oligodendrocyte progenitor cells (OPCs) from NMRs and compared their transcriptome with that of other mammals. Extracellular matrix (ECM) genes best distinguish OPCs of long- and short-lived species. Notably, expression levels of CD44, an ECM-binding protein that has been suggested to contribute to NMR longevity by mediating the effect of hyaluronan (HA), are not only high in OPCs of long-lived species but also positively correlate with longevity in multiple cell types/tissues. We found that CD44 localizes to the endoplasmic reticulum (ER) and enhances basal ATF6 activity. CD44 modifies proteome and membrane properties of the ER and enhances ER stress resistance in a manner dependent on unfolded protein response regulators without the requirement of HA. HA-independent role of CD44 in proteostasis regulation may contribute to mammalian longevity.

Opportunistic availability of an infant and social relationship to a mother influence grooming before infant handling in Japanese macaques.

Biological market theory (BMT) predicts that the balance between supply and demand determines the economic value of a "commodity," which consequently determines the amount of "services" that an individual needs to provide to obtain the commodity. In infant handling among primates, the existing literature suggests that handlers need to groom a mother to obtain access to its infant, particularly when the value of the infant is high (e.g., when the number of infants is low). However, grooming by handlers may not be a prerequisite for the occurrence of infant handling, because handlers can handle an infant that is separated from its mother. Based on 3 years of behavioral observations of wild Japanese macaques (Macaca fuscata), we examined the process by which infant handling occurs and the role of grooming in infant handling. We found that infant handling occurred more frequently when the mother and infant were separated than when they were in contact. Grooming rarely occurred before infant handling. The subsequent occurrence of infant handling was not predicted by either the occurrence or duration of grooming toward mothers by non-mother individuals. Grooming by handlers was also more likely to occur when an infant was in contact with its mother and when an infant's mother was dominant to the handlers. Contrary to BMT, the number of infants in a group did not affect grooming by handlers. These results indicate that the handlers' decision to groom depended on the opportunistic availability of an infant and the social relationship between the infant's mother and themselves. We conclude that grooming was not always necessary for infant handling.

Contagious yawning in African painted dogs.

Contagious yawning (CY), which is yawning elicited by sensing another yawning, has been observed only in social species and is considered linked to high sociality. Although this idea-the social communication hypothesis-is supported by previous studies, investigating the occurrence of CY in various species remains necessary. We investigated the occurrence of CY in one of the most social canine species, the African painted dog (Lycaon pictus). We recorded 1387 yawn events from five pairs (10 individuals) in captivity. Temporal analysis showed that subsequent yawns occurred frequently within 15 s or 30 s after spontaneous yawns (SYs). SYs that were detectable by another individual (i.e., visible to the other individual or performed in close proximity) were more likely to elicit subsequent yawns by the other individual. However, the influence of contextual factors on CY differed according to the time window, implying that a conservative time window should be used to reduce overcounting of CY and misattribution of its determinants. The proportion of CY was positively related to time spent in physical proximity to another, a proxy for a social bond. Overall, our results provide the first evidence of CY in African painted dogs and further support the notion that such behavior is prevalent among social animals. They also strongly imply that an appropriate time window should be used to define yawn contagion.

Development of Genetic Markers for Sex and Individual Identification of the Japanese Giant Flying Squirrel (Petaurista leucogenys) by an Efficient Method Using High-Throughput DNA Sequencing.

DNA markers that detect differences in the number of microsatellite repeats can be highly effective for genotyping individuals that lack differences in external morphology. However, isolation of sequences with different microsatellite repeat numbers between individuals has been a time-consuming process in the development of DNA markers. Individual identification of Japanese giant flying squirrels (Petaurista leucogenys) has been challenging because this species is arboreal and nocturnal and exhibits little to no morphological variation between individuals. In this study, we developed DNA markers for sex and individual identification of this species by an efficient method using high-throughput DNA sequence data. Paired-end 5 Gb (2 × 250 bp) and 15 Gb (2 × 150 bp) genome sequences were determined from a female and a male Japanese giant flying squirrel, respectively. We searched SRY and XIST genes located on Y and X chromosomes, respectively, from high-throughput sequence data and designed primers to amplify these genes. Using these primer sets, we succeeded to identify the sex of individuals. In addition, we selected 12 loci containing microsatellites with different numbers of repeats between two individuals from the same data set, and designed primers to amplify these sequences. Twenty individuals from nine different locations were discriminated using these primer sets. Furthermore, both sex and microsatellite markers were amplified from DNA extracted non-invasively from single fecal pellet samples. Based on our results for flying squirrels, we expect our efficient method for developing non-invasive high-resolution individual- and sex-specific genotyping to be applicable to a diversity of mammalian species.

A novel phylogenetic comparative method for evaluating the strength of branch-specific directional selection.

Phylogenetic comparative methods (PCMs) have played a central role in studying the evolution of phenotypic traits. However, when a trait experienced directional selection, previous PCMs have faced a dilemma between mathematically tractable but restrictive models (i.e., simple Gaussian process models) and flexible but intractable approaches (i.e., a simulation-based process model of phenotype evolution built on population genetics frameworks). This paper proposes a novel Gaussian process macroevolutionary model, called the "branch-specific directional selection (BSDS)," for evaluating the strength of directional selection to reconcile these two approaches. This model is based on a second-order approximation of a previous simulation-based process model but has a closed-form likelihood function. This can also be extended to incorporate intraspecies variations and to linear mixed models, which are necessary for meta-analysis. We conduct numerical experiments to validate the proposed method and apply it to the brain volume of Hominidae species. The results show that the proposed methods yield statistically more reliable inferences and computational time is about hundred thousand times faster than the previous simulation-based methods. Further extensions of the BSDS model are expected to provide a clearer picture of the connection of microevolutionary processes and macroevolutionary patterns.

Play face in Japanese macaques reflects the sender's play motivation.

Animals often initiate social interactions by exchanging signals. Especially when initiating amicable interactions, signaling one's friendly stance toward others in advance may be important to avoid being misunderstood as having hostile intentions. We used data on dyadic play fighting in a group of Japanese macaques, Macaca fuscata, to examine the function of "play face" at the opening of a play session. We found no support for the previously proposed hypothesis that play face expression is likelier before entering risky situations (e.g., before gaining an undue advantage over the partner) to avoid being misunderstood. The results showed that play face expression was likelier in male juveniles before initiating play with other males than in females before initiating play with males or other females and that juveniles were likelier to express play face before initiating play with others closer in age. As male Japanese macaques play more frequently than females, and juveniles prefer to play with individuals closer in age, play face expression before play initiation may reflect the individual's motivation for subsequent play interactions. This interpretation is supported by our observation that play bouts lasted longer when initiated with bidirectional play face by both participants than when initiated without play face. We also argued that since there was no tendency that play face was likelier to be expressed toward individuals with low play propensity (e.g., females) or infrequent partners to play with (e.g., individuals more distant in age), Japanese macaques may not tactically deploy this signal to recruit reluctant partners.

The Naked Mole-Rat as a Model for Healthy Aging.

Naked mole-rats (NMRs, Heterocephalus glaber) are the longest-lived rodents with a maximum life span exceeding 37 years. They exhibit a delayed aging phenotype and resistance to age-related functional decline/diseases. Specifically, they do not display increased mortality with age, maintain several physiological functions until nearly the end of their lifetime, and rarely develop cancer and Alzheimer's disease. NMRs live in a hypoxic environment in underground colonies in East Africa and are highly tolerant of hypoxia. These unique characteristics of NMRs have attracted considerable interest from zoological and biomedical researchers. This review summarizes previous studies of the ecology, hypoxia tolerance, longevity/delayed aging, and cancer resistance of NMRs and discusses possible mechanisms contributing to their healthy aging. In addition, we discuss current issues and future perspectives to fully elucidate the mechanisms underlying delayed aging and resistance to age-related diseases in NMRs.

Plasticity for the kin and conspecific preferences in the frog tadpoles (Rana ornativentris).

In vertebrates, little is known of kin recognition systems and their plasticity. Even in well-studied anuran larvae (tadpoles), the determinants and effects of prior experience have not been clarified. This study evaluates the plasticity of kin and conspecific discrimination in tadpoles of the Japanese montane brown frog Rana ornativentris. We raised tadpoles under two different sibship conditions: the pure line, comprising only siblings, and the mixed line, comprising both siblings and non-siblings. The association preference by a subject tadpole to unfamiliar ("stimulus") tadpoles was assessed through binary-choice tests using a 2 × 2 × 2 factorial design among each kinship line (pure and mix), subject ontogeny/size (early stage/small and late-stage/large), and stimuli ontogeny/size. Contrary to our expectations, kin preference was confirmed only in early developmental small tadpoles from mixed line, and only with a small stimulus. Furthermore, tadpoles from mixed line did not exhibit size preference for unrelated conspecifics. These results suggest that different prior associations have modulated kin templates along tadpole ontogeny and that the presence of non-kin would enhance the learning of kin/non-kin. This study provides the first example that plasticity of kin recognition affects not only kin-biased association but also conspecific recognition along ontogeny in tadpoles.

The naked truth: a comprehensive clarification and classification of current 'myths' in naked mole-rat biology.

The naked mole-rat (Heterocephalus glaber) has fascinated zoologists for at least half a century. It has also generated considerable biomedical interest not only because of its extraordinary longevity, but also because of unusual protective features (e.g. its tolerance of variable oxygen availability), which may be pertinent to several human disease states, including ischemia/reperfusion injury and neurodegeneration. A recent article entitled 'Surprisingly long survival of premature conclusions about naked mole-rat biology' described 28 'myths' which, those authors claimed, are a 'perpetuation of beautiful, but falsified, hypotheses' and impede our understanding of this enigmatic mammal. Here, we re-examine each of these 'myths' based on evidence published in the scientific literature. Following Braude et al., we argue that these 'myths' fall into four main categories: (i) 'myths' that would be better described as oversimplifications, some of which persist solely in the popular press; (ii) 'myths' that are based on incomplete understanding, where more evidence is clearly needed; (iii) 'myths' where the accumulation of evidence over the years has led to a revision in interpretation, but where there is no significant disagreement among scientists currently working in the field; (iv) 'myths' where there is a genuine difference in opinion among active researchers, based on alternative interpretations of the available evidence. The term 'myth' is particularly inappropriate when applied to competing, evidence-based hypotheses, which form part of the normal evolution of scientific knowledge. Here, we provide a comprehensive critical review of naked mole-rat biology and attempt to clarify some of these misconceptions.

The presence of females induces elevated cortisol levels in an alpha male: Experimental evidence in chimpanzees.

OBJECTIVES: In group-living primates, it has been reported that the alpha male exhibits high concentrations of cortisol and testosterone in the context of mating competition. We investigated how the presence of females affected salivary cortisol and testosterone levels in males from a small captive group of chimpanzees (Pan troglodytes). Specifically, we assessed whether the presence of females resulted in a rapid increase in salivary cortisol and testosterone levels in the alpha male. MATERIALS AND METHODS: We compared the social behavior and salivary hormone concentrations of four males before and after the presentation of receptive females. Three times a day, we collected saliva samples, a useful matrix for investigating short-term hormonal changes, and measured cortisol and testosterone concentration by liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: The frequency of inter-male aggression increased in the presence of females, indicating intense competition among males. Salivary cortisol levels increased in all males in the presence of females; however, the increase was significantly more pronounced in the alpha male. We found a complex three-way interaction among the presence of females, sampling timings, and male dominance rank in the analysis of salivary testosterone. Contrary to our prediction, a post hoc analysis revealed that salivary testosterone levels decreased after female introduction and that the alpha male did not show a higher level of salivary testosterone. CONCLUSIONS: Our study provides experimental evidence suggesting that the presence of females plays a significant role in the rank-related variation in the cortisol levels in male chimpanzees. Furthermore, our findings demonstrate the usefulness of salivary hormones for detecting short-term physiological changes in studies of socioendocrinology.

Directional selection in the evolution of elongated upper canines in clouded leopards and sabre-toothed cats.

Extremely developed or specialized traits such as the elongated upper canines of extinct sabre-toothed cats are often not analogous to those of any extant species, which limits our understanding of their evolutionary cause. However, an extant species may have undergone directional selection for a similar extreme phenotype. Among living felids, the clouded leopard, Neofelis nebulosa, has exceptionally long upper canines for its body size. We hypothesized that directional selection generated the elongated upper canines of clouded leopards in a manner similar to the process in extinct sabre-toothed cats. To test this, we developed an approach that compared the effect of directional selection among lineages in a phylogeny using a simulation of trait evolution and approximate Bayesian computation. This approach was applied to analyse the evolution of upper canine length in the Felidae phylogeny. Our analyses consistently showed directional selection favouring longer upper canines in the clouded leopard lineage and a lineage leading to the sabre-toothed cat with the longest upper canines, Smilodon. Most of our analyses detected an effect of directional selection for longer upper canines in the lineage leading to another sabre-toothed cat, Homotherium, although this selection may have occurred exclusively in the primitive species. In all the analyses, the clouded leopard and Smilodon lineages showed comparable directional selection. This implies that clouded leopards share a selection advantage with sabre-toothed cats in having elongated upper canines.

Redirected aggression as a conflict management tactic in the social cichlid fish Julidochromis regani.

Conflict management consists of social behaviours that reduce the costs of conflict among group members. Redirected aggression-that is, when a recently attacked individual attacks a third party immediately after the original aggression-is considered a conflict management tactic, as it may reduce the victim's probability of being the object of further aggression. Redirected aggression has been reported in many vertebrates, but few quantitative studies have been conducted on this behaviour in fishes. We examined the function of redirected aggression in Julidochromis regani, a social cichlid fish. Behavioural experiments showed that redirected aggression functioned to divert the original aggressor's attention towards a third party and to pre-empt an attack towards the victim by the third-party individual, specifically among females. We found, however, that redirected aggression did not delay the recurrence of aggression by the original aggressor. These results suggest that a primary function of redirected aggression is to maintain the dominance of its actor against a subordinate occupying an adjacent rank. This study provides, to our knowledge, the first evidence that redirected aggression functions to manage conflict in social fish.

Foraging intention affects whether willow tits call to attract members of mixed-species flocks.

Understanding how individual behaviour influences the spatial and temporal distribution of other species is necessary to resolve the complex structure of species assemblages. Mixed-species bird flocks provide an ideal opportunity to investigate this issue, because members of the flocks are involved in a variety of behavioural interactions between species. Willow tits (Poecile montanus) often produce loud calls when visiting a new foraging patch to recruit other members of mixed-species flocks. The costs and benefits of flocking would differ with individual foraging behaviours (i.e. immediate consumption or caching); thus, willow tits may adjust the production of loud calls according to their foraging intention. In this study, we investigated the link between foraging decisions and calling behaviour in willow tits and tested its influence on the temporal cohesion with members of mixed-species flocks. Observations at experimental foraging patches showed that willow tits produced more calls when they consumed food items compared with when they cached them. Playback experiments revealed that these calls attracted flock members and helped to maintain their presence at foraging patches. Thus, willow tits adjusted calling behaviour according to their foraging intention, thereby coordinating the associations with members of mixed-species flocks. Our findings demonstrate the influence of individual decision-making on temporal cohesion with other species and highlight the importance of interspecific communication in mixed-species flocking dynamics.

Male-specific mortality biases secondary sex ratio in Eurasian tree sparrows Passer montanus.

Sex allocation theory predicts that parents bias the offspring sex ratio strategically. In avian species, the offspring sex ratio can be biased at multiple growth stages, although the mechanisms are not well known. It is crucial to reveal a cause and timing of biased offspring sex ratio. We investigated (i) offspring sex ratio at multiple growth stages, from laying to fledging; and (ii) the stage at which offspring sex ratio became biased; and (iii) the cause of biased offspring sex ratio in Eurasian tree sparrows Passer montanus. Sex determination of 218 offspring, including hatchlings and unhatched eggs from 41 clutches, suggested that the offspring sex ratio was not biased at the egg-laying stage but was significantly female-biased after the laying stage due to higher mortality of male embryos. Half of the unhatched eggs showed no sign of embryo development (37/74, 50.00%), and most undeveloped eggs were male (36/37, 97.30%). Additional experiments using an incubator suggested that the cause of embryo developmental failure was a lack of developmental ability within the egg, rather than a failure of incubation. This study highlights the importance of clarifying offspring sex ratio at multiple stages and suggests that offspring sex ratio is adjusted after fertilization.

Evolution of tail fork depth in genus Hirundo.

A classic example of a sexually selected trait, the deep fork tail of the barn swallow Hirundo rustica is now claimed to have evolved and be maintained mainly via aerodynamic advantage rather than sexually selected advantage. However, this aerodynamic advantage hypothesis does not clarify which flight habits select for/against deep fork tails, causing diversity of tail fork depth in hirundines. Here, by focusing on the genus Hirundo, we investigated whether the large variation in tail fork depth could be explained by the differential flight habits. Using a phylogenetic comparative approach, we found that migrant species had deeper fork tails, but less colorful plumage, than the other species, indicating that migration favors a specific trait, deep fork tails. At the same time, tail fork depth but not plumage coloration decreased with increasing bill size - a proxy of prey size, suggesting that foraging on larger prey items favors shallower fork tails. Variation of tail fork depth in the genus Hirundo may be explained by differential flight habits, even without assuming sexual selection.

Asian elephants acquire inaccessible food by blowing.

Many animals acquire otherwise inaccessible food with the aid of sticks and occasionally water. As an exception, some reports suggest that elephants manipulate breathing through their trunks to acquire inaccessible food. Here, we report on two female Asian elephants (Elephas maximus) in Kamine Zoo, Japan, who regularly blew to drive food within their reach. We experimentally investigated this behaviour by placing foods in inaccessible places. The elephants blew the food until it came within accessible range. Once the food was within range, the elephants were increasingly less likely to blow as the distance to the food became shorter. One subject manipulated her blowing duration based on food distance: longer when the food was distant. These results suggest that the elephants used their breath to achieve goals: that is, they used it not only to retrieve the food but also to fine-tune the food position for easy grasping. We also observed individual differences in the elephants' aptitude for this technique, which altered the efficiency of food acquisition. Thus, we added a new example of spontaneous behaviour for achieving a goal in animals. The use of breath to drive food is probably unique to elephants, with their dexterous trunks and familiarity with manipulating the act of blowing, which is commonly employed for self-comfort and acoustic communication.

Head rubbing and licking reinforce social bonds in a group of captive African lions, Panthera leo.

Many social animals have a species-specific repertoire of affiliative behaviours that characterise individualised relationships within a group. To date, however, quantitative studies on intragroup affiliative behaviours in social carnivores have been limited. Here, we investigated the social functions of the two most commonly observed affiliative behaviours in captive African lions (Panthera leo): head rubbing and licking. We conducted behavioural observations on a captive group of lions composed of 7 males and 14 females, and tested hypotheses regarding three social functions: tension reduction, social bonding, and social status expression. Disproportionately frequent male-male and female-to-male head rubbing was observed, while more than 95% of all licking interactions occurred in female-female dyads. In accordance with the social bond hypothesis, and in disagreement with the social status expression hypothesis, both head rubbing and licking interactions were reciprocal. After controlling for spatial association, the dyadic frequency of head rubbing was negatively correlated with age difference while licking was positively correlated with relatedness. Group reunion after daily separation did not affect the frequencies of the affiliative behaviours, which was in disagreement with the predictions from the tension reduction hypothesis. These results support the social bond hypothesis for the functions of head rubbing and licking. Different patterns of affiliative behaviour between the sexes may reflect differences in the relationship quality in each sex or the differential predisposition to licking due to its original function in offspring care.

Simulation-based likelihood approach for evolutionary models of phenotypic traits on phylogeny.

Phylogenetic comparative methods (PCMs) have been used to test evolutionary hypotheses at phenotypic levels. The evolutionary modes commonly included in PCMs are Brownian motion (genetic drift) and the Ornstein-Uhlenbeck process (stabilizing selection), whose likelihood functions are mathematically tractable. More complicated models of evolutionary modes, such as branch-specific directional selection, have not been used because calculations of likelihood and parameter estimates in the maximum-likelihood framework are not straightforward. To solve this problem, we introduced a population genetics framework into a PCM, and here, we present a flexible and comprehensive framework for estimating evolutionary parameters through simulation-based likelihood computations. The method does not require analytic likelihood computations, and evolutionary models can be used as long as simulation is possible. Our approach has many advantages: it incorporates different evolutionary modes for phenotypes into phylogeny, it takes intraspecific variation into account, it evaluates full likelihood instead of using summary statistics, and it can be used to estimate ancestral traits. We present a successful application of the method to the evolution of brain size in primates. Our method can be easily implemented in more computationally effective frameworks such as approximate Bayesian computation (ABC), which will enhance the use of computationally intensive methods in the study of phenotypic evolution.