Shifts in energy allocation and reproduction in response to temperature in a small precocial mammal.

BACKGROUND: Species adjust to changes in temperature and the accompanying reduction in resource availability during the annual cycle by shifts in energy allocation. As it gets colder, the priority of energy allocation to maintenance increases and reproduction is reduced or abandoned. RESULTS: We studied whether and how young female guinea pigs (Cavia porcellus) adjust even under ad libitum food conditions growth, storage of fat reserves and reproduction when kept at 5 °C versus 15 °C, and how offspring born into these conditions compensate during development to independence. Reproducing females grew less in the cold. Their lower weight resulted largely from less fat storage whereas growth in fat free mass was about the same for both groups. The increased need for thermoregulation diminished fat storage most likely due to the development of more brown fat tissue. Reproductive activity did not differ between groups in terms of litter frequency, mass and size. However, females in 5 °C weaned pups later (around day 25) than females in 15 °C (around day 21). Later weaning did not make up for the higher energy expenditure of pups in cold conditions leading to slower growth and less fat storage. Female pups born into the cold matured later than those born in 15 °C. Investment in reproduction continued but allocation to individual pups declined. CONCLUSIONS: In more thermally demanding conditions female guinea pigs - even under ad libitum food abundance - transfer the higher costs of maintenance and reproduction largely to offspring.

Stressfulness of the design influences consistency of cognitive measures and their correlation with animal personality traits in wild mice (Mus musculus).

Individual variation in cognition is being increasingly recognized as an important evolutionary force but contradictory results so far hamper a general understanding of consistency and association with other behaviors. Partly, this might be caused by external factors imposed by the design. Stress, for example, is known to influence cognition, with mild stress improving learning abilities, while strong or chronic stress impairs them. Also, there might be intraspecific variation in how stressful a given situation is perceived. We investigated two personality traits (stress coping and voluntary exploration), spatial learning with two mazes, and problem-solving in low- and high-stress tests with a group of 30 female wild mice (Mus musculus domesticus). For each test, perceived stress was assessed by measuring body temperature change with infrared thermography, a new non-invasive method that measures skin temperature as a proxy of changes in the sympathetic system activity. While spatial learning and problem-solving were found to be repeatable traits in mice in earlier studies, none of the learning measures were significantly repeatable between the two stress conditions in our study, indicating that the stress level impacts learning. We found correlations between learning and personality traits; however, they differed between the two stress conditions and between the cognitive tasks, suggesting that different mechanisms underlie these processes. These findings could explain some of the contradictory findings in the literature and argue for very careful design of cognitive test setups to draw evolutionary implications.

Fast adjustment of pace-of-life and risk-taking to changes in food quality by altered gene expression in house mice.

The pace-of-life syndrome hypothesis provides a framework for the adaptive integration of behaviour, physiology and life history between and within species. It suggests that behaviours involving a risk of death or injury should co-vary with a higher allocation to fast reproduction. Empirical support for this hypothesis is mixed, presumably because important influencing factors such as environmental variation, are usually neglected. By experimentally manipulating food quality of wild mice living under semi-natural conditions for three generations, we show that individuals adjust their life history strategies and risk-taking behaviours as well as trait covariation (Nindividuals  = 1442). These phenotypic differences are correlated to differences in transcriptomic gene expression of primary metabolic processes in the liver while no changes in gene frequencies occurred. Our discussion emphasises the need to integrate the role of environmental conditions and phenotypic plasticity in shaping relationships among behaviour, physiology and life history in response to changing environmental conditions.

Lifetime Observation of Cognition and Physiological Parameters in Male Mice.

Laboratory mice are predominantly used for one experiment only, i.e., new mice are ordered or bred for every new experiment. Moreover, most experiments use relatively young mice in the range of late adolescence to early adulthood. As a consequence, little is known about the day-to-day life of adult and aged laboratory mice. Here we present a long-term data set with three consecutive phases conducted with the same male mice over their lifetime in order to shed light on possible long-term effects of repeated cognitive stimulation. One third of the animals was trained by a variety of learning tasks conducted up to an age of 606 days. The mice were housed in four cages with 12 animals per cage; only four mice per cage had to repeatedly solve cognitive tasks for getting access to water using the IntelliCage system. In addition, these learner mice were tested in standard cognitive tests outside their home-cage. The other eight mice served as two control groups living in the same environment but without having to solve tasks for getting access to water. One control group was additionally placed on the test set-ups without having to learn the tasks. Next to the cognitive tasks, we took physiological measures (body mass, resting metabolic rate) and tested for dominance behavior, and attractivity in a female choice experiment. Overall, the mice were under surveillance until they died a natural death, providing a unique data set over the course of virtually their entire lives. Our data showed treatment differences during the first phase of our lifetime data set. Young learner mice showed a higher activity, less growth and resting metabolic rate, and were less attractive for female mice. These effects, however, were not preserved over the long-term. We also did not find differences in dominance or effects on longevity. However, we generated a unique and valuable set of long-term behavioral and physiological data from a single group of male mice and note that our long-term data contribute to a better understanding of the behavioral and physiological processes in male C57Bl/6J mice.

Enhanced problem-solving ability as an adaptation to urban environments in house mice.

Humans have a large impact on the distribution and abundance of animal species worldwide. The ecological effects of human-altered environments are being increasingly recognized and understood, but their effects on evolution are largely unknown. Enhanced cognitive abilities and the ability to innovate have been suggested as crucial traits for thriving in human-altered habitats. We tested if house mice (Mus musculus) subspecies have evolved enhanced innovative problem-solving abilities throughout their commensal lives with humans. The time that subspecies lived commensally with humans ranges between approximately 3000 years to more than 11 000 years, thus providing an excellent example of human-animal coexistence. In addition, we tested whether differences in problem-solving were mediated by differences in object and place exploration, motivation, persistence or inhibitory control. We found that populations of subspecies living commensally the longest excelled in problem-solving across seven food-extraction tasks over subspecies living commensally short or intermediate times. These differences were not mediated by exploration, motivation, persistence or inhibitory control suggesting that subspecies have evolved better cognitive abilities when living commensally in urban environments. This suggests that the ability to problem-solve may be an important trait promoting prosperity in human-altered environments.

Zebra finches bi-directionally selected for personality differ in repeatability of corticosterone and testosterone.

Consistent between-individual differences in behaviour have been documented across the animal kingdom. Such variation between individuals has been shown to be the basis for selection and to act as a pacemaker for evolutionary change. Recently, equivocal evidence suggests that such consistent between-individual variation is also present in hormones. This observation has sparked interest in understanding the mechanisms shaping individual differences, temporal consistency and heritability of hormonal phenotypes and to understand, if and to what extent hormonal mechanisms are involved in mediating consistent variation in behaviour between individuals. Here, we used zebra finches of the fourth generation of bi-directionally selected lines for three independent behaviours: aggression, exploration and fearlessness. We investigated how these behaviours responded to artificial selection and tested their repeatability. We further tested for repeatability of corticosterone and testosterone across and within lines. Moreover, we are presenting the decomposed variance components for within-individual variance (i.e. flexibility) and between-individual variance (i.e. more or less pronounced differences between individuals) and investigate their contribution to repeatability estimates. Both hormones as well as the exploration and fearlessness but not aggressiveness, were repeatable. However, variance components and hence repeatability differed between lines and were often lower than in unselected control animals, mainly because of a reduction in between-individual variance. Our data show that artificial selection (including active selection and genetic drift) can affect the mean and variance of traits. We stress the importance for understanding how variable a trait is both between and within individuals to assess the selective value of a trait.

Within-Family Environment and Cross-Fostering Stress Affect Behavior and Physiology in Wild Cavies (Cavia aperea).

Stability of personality traits is well-documented for a wide variety of animals. However, previous results also suggest that behavioral phenotypes are plastic during early ontogeny and can be adaptively shaped to the social environment. In cavies (Cavia aperea), it has already been documented that the size at birth relative to siblings (size rank) greatly influences various behavioral and physiological traits that last at least until independence. The aim of the current study was (1) to investigate if behavioral and physiological differences between pups of the same litter persist until after independence and influence development long-lasting, (2) to determine the potential plasticity in response to changes in the early within-family environment by cross-fostering pups either to the same, a lower, or a higher size rank in a foster-family. We measured three behavioral traits (number of interactions with a novel object, distance moved in an open field, struggle docility) and two physiological traits (resting metabolic rate and basal cortisol levels). We predicted that cross-fostering into a litter where pups occupy the same size rank would not change the expression of traits. Cross-fostering to a different size rank should not influence the expression of traits if repeatability measures indicate low plasticity. Alternatively, if the traits are plastic, animals should adjust trait expression to fit with the size rank occupied in the foster litter. Initial differences in struggle docility, distance moved in an open field and in baseline cortisol concentration between pups of different size-ranks did not remain stable beyond independence. In addition, we found remarkable plasticity of the measured traits in response to cross-fostering to the same, a smaller or larger size-rank, suggesting that differences between pups are more the result of social constraints leading to adaptive shaping of individual phenotypes within a family. We also found a significant influence of the cross-fostering procedure itself. Cross-fostered individuals were less bold, grew slower and showed elevated resting metabolic rates. This finding suggests a cautious interpretation of previous cross-fostering studies and stresses the need for proper control groups to reliably separate the effect of cross-fostering per se from those induced by an experimental treatment.

Reproduction affects immune defenses in the guinea pig even under ad libitum food.

Reproduction is one of the costliest processes in the life of an animal. Life history theory assumes that when resources are limiting allocation to reproduction will reduce allocation to other essential processes thereby inducing costs of reproduction. The immune system is vital for survival. If reproduction reduces investment in immune function, this could increase the risk of disease, morbidity and mortality. We here test in the guinea pig, if even under ad libitum food conditions, pregnancy and lactation reduce the activity of the adaptive and innate immune system compared to the reaction of non-reproducing animals. In response to a challenge with keyhole limpet haemocyanin the antibody-mediated adaptive immunity during (pregnancy and) lactation was reduced. Pregnant and lactating females showed higher levels of bacterial killing activity, an integrated measure of innate immunity, than non-reproducing females. However, two major effectors of the innate immunity, the natural antibody and the complement of pregnant and lactating females showed lower levels than in non-reproducing females. Pregnant and lactating females did not differ significantly in the expressed levels of innate immunity. Our results indicate that changes in the immune response during reproduction are physiological adjustments to predictable allocation problems, because they happen even under ad libitum food availability.

Senescence and costs of reproduction in the life history of a small precocial species.

Species following a fast life history are expected to express fitness costs mainly as increased mortality, while slow-lived species should suffer fertility costs. Because observational studies have limited power to disentangle intrinsic and extrinsic factors influencing senescence, we manipulated reproductive effort experimentally in the cavy (Cavia aperea) which produces extremely precocial young. We created two experimental groups: One was allowed continuous reproduction (CR) and the other intermittent reproduction (IR) by removing males at regular intervals. We predicted that the CR females should senesce (and die) earlier and produce either fewer and/or smaller, slower growing offspring per litter than those of the IR group. CR females had 16% more litters during three years than IR females. CR females increased mass and body condition more steeply and both remained higher until the experiment ended. Female survival showed no group difference. Reproductive senescence in litter size, litter mass, and reproductive effort (litter mass/maternal mass) began after about 600 days and was slightly stronger in CR than IR females. Litter size, litter mass, and offspring survival declined with maternal age and were influenced by seasonality. IR females decreased reproductive effort less during cold seasons and only at higher age than CR females. Nevertheless, offspring winter mortality was higher in IR females. Our results show small costs of reproduction despite high reproductive effort, suggesting that under ad libitum food conditions costs depend largely on internal regulation of allocation decisions.

Does Animal Personality Affect Movement in Habitat Corridors? Experiments with Common Voles (Microtus arvalis) using Different Corridor Widths.

Animal personality may affect an animal's mobility in a given landscape, influencing its propensity to take risks in an unknown environment. We investigated the mobility of translocated common voles in two corridor systems 60 m in length and differing in width (1 m and 3 m). Voles were behaviorally phenotyped in repeated open field and barrier tests. Observed behavioral traits were highly repeatable and described by a continuous personality score. Subsequently, animals were tracked via an automated very high frequency (VHF) telemetry radio tracking system to monitor their movement patterns in the corridor system. Although personality did not explain movement patterns, corridor width determined the amount of time spent in the habitat corridor. Voles in the narrow corridor system entered the corridor faster and spent less time in the corridor than animals in the wide corridor. Thus, landscape features seem to affect movement patterns more strongly than personality. Meanwhile, site characteristics, such as corridor width, could prove to be highly important when designing corridors for conservation, with narrow corridors facilitating faster movement through landscapes than wider corridors.

An Autaptic Culture System for Standardized Analyses of iPSC-Derived Human Neurons.

iPSC-derived human neurons are expected to revolutionize studies on brain diseases, but their functional heterogeneity still poses a problem. Key sources of heterogeneity are the different cell culture systems used. We show that an optimized autaptic culture system, with single neurons on astrocyte feeder islands, is well suited to culture, and we analyze human iPSC-derived neurons in a standardized, systematic, and reproducible manner. Using classically differentiated and transcription factor-induced human glutamatergic and GABAergic neurons, we demonstrate that key features of neuronal morphology and function, including dendrite structure, synapse number, membrane properties, synaptic transmission, and short-term plasticity, can be assessed with substantial throughput and reproducibility. We propose our optimized autaptic culture system as a tool to study functional features of human neurons, particularly in the context of disease phenotypes and experimental therapy.

Future Directions for Personality Research: Contributing New Insights to the Understanding of Animal Behavior.

As part of the European Conference on Behavioral Biology 2018, we organized a symposium entitled, "Animal personality: providing new insights into behavior?" The aims of this symposium were to address current research in the personality field, spanning both behavioral ecology and psychology, to highlight the future directions for this research, and to consider whether differential approaches to studying behavior contribute something new to the understanding of animal behavior. In this paper, we discuss the study of endocrinology and ontogeny in understanding how behavioral variation is generated and maintained, despite selection pressures assumed to reduce this variation. We consider the potential mechanisms that could link certain traits to fitness outcomes through longevity and cognition. We also address the role of individual differences in stress coping, mortality, and health risk, and how the study of these relationships could be applied to improve animal welfare. From the insights provided by these topics, we assert that studying individual differences through the lens of personality has provided new directions in behavioral research, and we encourage further research in these directions, across this interdisciplinary field.

Sex differences in life history, behavior, and physiology along a slow-fast continuum: a meta-analysis.

The pace-of-life syndrome (POLS) hypothesis predicts that behavior and physiology covary with life history. Evidence for such covariation is contradictory, possibly because systematic sources of variation (e.g. sex) have been neglected. Sexes often experience different selection pressures leading to sex-specific allocation between reproduction and self-maintenance, facilitating divergence in life-history. Sex-specific differences in means and possibly variances may therefore play a key role in the POLS framework. We investigate whether sexes differ in means and variances along the fast-slow pace-of-life continuum for life history and physiological and behavioral traits. In addition, we test whether social and environmental characteristics such as breeding strategy, mating system, and study environment explain heterogeneity between the sexes. Using meta-analytic methods, we found that populations with a polygynous mating system or for studies conducted on wild populations, males had a faster pace-of-life for developmental life-history traits (e.g., growth rate), behavior, and physiology. In contrast, adult life-history traits (e.g., lifespan) were shifted towards faster pace-of-life in females, deviating from the other trait categories. Phenotypic variances were similar between the sexes across trait categories and were not affected by mating system or study environment. Breeding strategy did not influence sex differences in variances or means. We discuss our results in the light of sex-specific selection that might drive sex-specific differences in pace-of-life and ultimately POLS.

Effect of season on reproductive behaviors and fertilization success in cavies (Cavia aperea).

Finding the optimal timing for breeding is crucial for small mammals to ensure survival and maximize lifetime reproductive success. Species living in temperate regions therefore often restrict breeding to seasons with favorable food and weather conditions. Although caviomorph rodents such as guinea pigs are described as non-seasonal breeders, a series of recent publications has shown seasonal adaptations in litter size, offspring birth mass and maternal investment. Here, we aim to test if seasonal patterns of litter size variation found in earlier studies, are mediated by seasonal differences in female estrus length, fertilization rate and mating behavior. The female estrus period was longer in fall compared to all other seasons (p < 0.001), frequently lasting 7-9 days while estrus in spring usually lasted less than 2 days. In fall, females mated later during estrus (p < 0.001), resulting in reduced fertilization rates (p < 0.001). Fertilization rate was well above 95% in summer while it dropped to less than 85% in fall and winter. While none of the male mating characteristics such as number and duration of copulations differed across seasons, the number of mating bouts was reduced in fall (p = 0.04). Finally, the developmental stages of flushed embryos were more diverse in spring and summer compared to fall and winter. These results suggest that seasonal differences in fertilization rate and quality of implanted embryos are mediated by female estrus length and timing and intensity of mating behavior. Together, these effects contribute to the observed differences in litter size across seasons.

Life-history trade-offs: are they linked to personality in a precocial mammal (Cavia aperea)?

Life-history trade-offs are predicted to contribute to the maintenance of personality variation. Individuals with 'fast' lifestyles should develop faster, reproduce earlier and exhibit more risky behaviours. Evidence for such predicted links, however, remains equivocal. Here, I test how growth rate, timing of maturation, litter size and maternal effort correlate with exploration, boldness, fearlessness, docility and escape latency. I found several links that were predicted by recent theory while others were against theoretical predictions, e.g. fast-growing individuals were more fearful. Thus, while I found personality to be integrated with life history, I cannot fully support recent hypotheses aiming to explain such behaviour-life-history associations.

Stability of the guinea pigs personality - cognition - linkage over time.

In human psychological research, personality traits as well as cognitive traits are usually validated for both, their stability over time and contexts. While stability over time gives an estimate on how genetically fixated a trait can be, correlations across traits have the power to reveal linkages or trade - offs. In animals, these validations have widely been done for personality but not for cognitive traits. We tested guinea pigs in four consecutive discrimination tasks using four unique pairs of objects with two objects of the same form but different size in each pair. The same animals were tested twice each for three personality traits, i.e. boldness, aggression and sociopositive behaviour. The animals did not learn to "always choose the larger item" in the cognitive task but learned to discriminate the two objects of each stimulus pair anew, so that we did test for learning speed in four slightly different task setups. Performance over the four tasks was significantly repeatable as well as all tested personality traits. A stable linkage over time was found between sociopositive behaviour and learning performance, probably indicating an ecological relevance for a correlation between these two traits. Still, not all traits seem to be connected amongst each other, as in our case boldness and aggression are both not linked to individual learning performance. Future studies will hopefully further investigate the repeatability of various cognitive traits in several species and thus lead to a better understanding of the interdependence of personality and cognition. This will help to unravel which suites of traits facilitate individual life histories and hence improve our understanding of the emergence and maintenance of individual differences.

Blastocyst recovery and multifactorial gene expression analysis in the wild guinea pig (Cavia aperea).

The expression of specific developmentally important genes in preimplantation embryos is an accepted marker for unraveling the influence of single factors in studies that are mostly related to artificial reproduction techniques. Such studies, however, often reveal high levels of heterogeneity between single embryos, independently of the influence of factors of interest. A possible explanation for this variation could be the large variety of physiological and environmental factors to which early embryos are exposed and their ability to react to them. Here, we investigated several potentially important parameters of development at the same time, in blastocysts of the wild guinea pig (Cavia aperea) generated in vivo after natural mating. The optimal time for flushing fully developed blastocysts was between 123 and 126 hours after mating. The abundance of POU5F1 (P = 0.042), BAX (P < 0.001), SLC2A1 (P = 0.017), and DNMT3A (P < 0.001) mRNA changed significantly over time after mating. The number of sibling embryos present influenced STAT3 levels significantly (P = 0.02). Levels of BAX and POU5F1 were significantly affected by season (P = 0.03 and 0.04). The temporal pattern of SLC2A1 levels was significantly altered both after feeding a protein-deficient diet (P = 0.04) and temperature treatment (P = 0.04) of the sire. In addition, the identity of the father had a significant influence on POU5F1 (P = 0.049) and STAT3 (P < 0.001) mRNA abundances. These data report that the expression of specific genes in early embryos reflects the entire heterogeneity of their surroundings and that it is a plastic reaction toward a multifactorial environment.

The maternal social environment shapes offspring growth, physiology, and behavioural phenotype in guinea pigs.

BACKGROUND: Prenatal conditions influence offspring development in many species. In mammals, the effects of social density have traditionally been considered a detrimental form of maternal stress. Now their potential adaptive significance is receiving greater attention.Sex-specific effects of maternal social instability on offspring in guinea pigs (Cavia aperea f. porcellus) have been interpreted as adaptations to high social densities, while the effects of low social density are unknown. Hence, we compared morphological, behavioural and physiological development between offspring born to mothers housed either individually or in groups during the second half of pregnancy. RESULTS: Females housed individually and females housed in groups gave birth to litters of similar size and sex-ratios, and there were no differences in birth weight. Sons of individually-housed mothers grew faster than their sisters, whereas daughters ofgroup-housed females grew faster than their brothers, primarily due to an effect on growth of daughters. There were few effects on offspring behaviour. Baseline cortisol levels in saliva of pups on day 1 and day 7 were not affected, but we saw a blunted cortisol response to social separation on day 7 in sons of individually-housed females and daughters of group-housed females. The effects were consistent across two replicate experiments. CONCLUSIONS: The observed effects only partially support the adaptive hypothesis. Increased growth of daughters may be adaptive under high densities due to increasedfemale competition, but it is unclear why growth of sons is not increased under low social densities when males face less competition from older, dominant males. The differences in growth may be causally linked to sex-specific effects on cortisol response, although individual cortisol response and growth were not correlated, and various other mechanisms are possible. The observed sex-specific effects on early development are intriguing, yet the potential adaptive benefits and physiological mechanisms require further study.

Domestication effects on behavioural traits and learning performance: comparing wild cavies to guinea pigs.

The domestication process leads to a change in behavioural traits, usually towards individuals that are less attentive to changes in their environment and less aggressive. Empirical evidence for a difference in cognitive performance, however, is scarce. Recently, a functional linkage between an individual's behaviour and cognitive performance has been proposed in the framework of animal personalities via a shared risk-reward trade-off. Following this assumption, bolder and more aggressive animals (usually the wild form) should learn faster. Differences in behaviour may arise during ontogeny due to individual experiences or represent adaptations that occurred over the course of evolution. Both might singly or taken together account for differences in cognitive performance between wild and domestic lineages. To test for such possible linkages, we compared wild cavies and domestic guinea pigs, both kept in a university stock for more than 30 years under highly comparable conditions. Animals were tested in three behavioural tests as well as for initial and reversal learning performance. Guinea pigs were less bold and aggressive than their wild congeners, but learnt an association faster. Additionally, the personality structure was altered during the domestication process. The most likely explanation for these findings is that a shift in behavioural traits and their connectivity led to an altered cognitive performance. A functional linkage between behavioural and cognitive traits seems to exist in the proposed way only under natural selection, but not in animals that have been selected artificially over centuries.

Animal housing and welfare: effects of housing conditions on body weight and cortisol in a medium-sized rodent (Cavia aperea).

Rodents are the most abundant experimental nonhuman animals and are commonly studied under standard laboratory housing conditions. As housing conditions affect animals' physiology and behavior, this study investigated the effects of indoor and outdoor housing conditions on body weight and cortisol level of wild cavies, Cavia aperea. The changing housing condition strongly influenced both parameters, which are commonly used as indicators for animal welfare. The transfer from outdoor to indoor enclosures resulted in a body-weight loss of about 8%. In contrast, animals kept indoors showed a substantial weight gain of about 12% when they were transferred outdoors. These effects were reversible. To substantiate a connection between body-weight changes and the health states of the animals, blood basal cortisol concentrations were measured. Animals kept outdoors had significantly lower cortisol levels than did animals kept indoors. These results imply that indoor conditions have a direct effect on the animals' states. The physiological and metabolic consequences as well as potential welfare aspects should be taken into account when planning experimental work, especially on nondomestic animals.