Effects of early predation and social cues on the relationship between laterality and personality.

Individual differences in laterality and personality are expected to covary, as emotions are processed differently by the two hemispheres, and personality involves emotional behavior. Fish species are often used to investigate this topic due to the large variability in personality and laterality patterns. While some species show a positive relationship between lateralization strength and boldness, others show a negative relationship, and some show no relationship. A new way to assess the robustness of such a relationship is to manipulate both laterality and personality to examine how this affects their relationship. To this end, we conducted a fully factorial design experiment manipulating predation and group size during early development. Results showed that the strength of laterality was influenced by predation threat, while social tendency and boldness were influenced by group size. These findings suggest that early life conditions can have an impact on laterality and social behavior. The relationship between laterality and personality traits, while present, was heavily influenced by the specific trial conditions but not by the different developmental conditions. In summary, the relationship between laterality and behaviors appears to be context-dependent, yet resilient to early environmental manipulations.

The effect of light during embryonic development on laterality and exploration in Western Rainbowfish.

Several factors affect the development of lateralization such as hormones and light exposure during early development. Laterality also often correlates with other behavioral traits. To examine whether there is a common mechanism underlying the development of laterality and other behaviors, we manipulated laterality by exposing embryos of the Western rainbowfish (Melatotaenia australis) to light or continuous darkness during early development and determined whether a shift in laterality was associated with a change in behavior in a novel environment test at two different ages. We found that exposing eggs to darkness led to offspring that displayed significantly less lateralized behavior in the mirror test two weeks after hatching than offspring from eggs exposed to light. Interestingly, the effects of rearing condition were lost by 3 months of age. These data suggest that exposure to light can influence laterality very early in development, but such bias can be overwritten by developmental processes post-hatch. Moreover, our manipulation of laterality apparently had no influence on exploration suggesting independent causal mechanisms. The experimental manipulation of light exposure during development could be a useful tool for enhancing individuals with a specific laterality and behavioral traits to aid future research into the causes and consequences of laterality.

Plasticity in metabolism of maternal androgens in avian embryos.

Mothers can influence offspring phenotypes by transferring non-genetic information to the young, which provides them with a flexible tool to adjust the developmental trajectory of the young in fluctuating environments. Mothers can differentially deposit their resources in the same reproductive attempt in relation to the offspring position in the sibling hierarchy. However, whether embryos from different positions can be plastic in their response to the maternal signals, potentially leading to a mother-offspring conflict, is yet unclear. We used Rock pigeons (Columba livia), that lay two egg clutches where maternal androgen levels in second laid eggs at oviposition are higher than in first laid eggs, and investigated the plasticity of embryonic metabolism of maternal androgens. We experimentally elevated androstenedione and testosterone levels in first eggs to that present in second eggs and measured the change in androgen levels and its main metabolites (etiocholanolone and conjugated testosterone) after 3.5 days of incubation. We found that eggs with increased androgens show a different degree of androgen metabolism depending either on the egg laying sequence or initial androgen levels or both. Our findings indicate that embryos have certain plasticity in response to maternal androgen levels depending on maternal signals.

Dynamics of maternal androgens and its metabolites during early embryonic development: embryonic modification of a maternal effect.

In birds, exposure to maternal (yolk) testosterone affects a diversity of offspring post-hatching traits, which eventually affect offspring competitiveness. However, maternal testosterone is heavily metabolized at very early embryonic developmental stages to hydrophilic metabolites that are often assumed to be much less biologically potent. Either the rapid metabolism could either keep the maternal testosterone from reaching the embryos, opening the possibility for a mother-offspring conflict or the metabolites may facilitate the uptake of the lipophilic testosterone from the yolk into the embryonic circulation after which they are either converted back to the testosterone or functioning directly as metabolites. To test these possibilities, we injected isotope-labeled testosterone (T-[D5]) into the yolk of freshly laid Rock pigeon (Columba livia) eggs and determined the concentration and distribution of T-[D5] and its labeled metabolites within different egg fractions by liquid chromatography combined with tandem mass spectrometry at day 2, 5 and 10 of incubation. Although under a supraphysiological dosage injection, yolk testosterone decreased within 2 days and was metabolized into androstenedione, conjugated testosterone, etiocholanolone and other components that were unidentifiable due to methodological limitation. We show for the first time that testosterone, androstenedione and conjugated testosterone, but not etiocholanolone, reached the embryo including its brain. Their high concentrations in the yolk and extraembryonic membranes suggest that conversion takes place here. We also found no sex-specific metabolism, explaining why maternal testosterone does not affect sexual differentiation. Our findings showed that maternal testosterone is quickly converted by the embryo, with several but not all metabolites reaching the embryo providing evidence for both hypotheses.

Early conversion of maternal androgens affects the embryo already in the first week of development.

Maternal androgen exposure has potent effects on offspring development. As substantial levels of maternal androgens are deposited in avian egg yolks, avian eggs are frequently used to study maternal effects, with a strong focus on post-natal development. However, the underlying pathways are largely unknown. Since the hormones are taken up during the embryonic phase, and these are rapidly metabolized by avian embryos into metabolites such as etiocholanolone, we studied the effects of yolk androgens (testosterone and androstenedione) and their metabolite etiocholanolone during the first few days of embryonic development. As embryonic heart rate is often used as an indicator of embryonic development, we measured the heart rate from day 3 to day 6 of incubation by using a shell-less culture technique in rock pigeon eggs (Columba livia). Increased androgen exposure increased heart rate, and increased etiocholanolone mimicked this effect, albeit in a small sample size. This indicates that exposure to maternal androgens increases embryonic overall metabolism which may account for the developmental outcomes found in previous studies such as increased growth. Moreover, etiocholanolone is likely to be an important metabolite in a non-genomic pathway underlying the androgen-mediated maternal effect.

Embryonic heart rate is affected by yolk androgens and egg laying sequence, and correlates with embryonic tissue growth: A study in rock pigeons.

Maternal androgen exposure can have crucial effects on offspring development. Bird eggs are frequently used for studying these effects and virtually all research in this field has focused on post-hatching offspring traits. Yet, much of the yolk, in which the maternal hormones are deposited, is consumed during the embryonic phase. Here, we studied the effects of yolk androgens during this prenatal period. As there is evidence that androgens stimulate post-hatching traits such as increased growth, we measured heart rate throughout incubation as a proxy for prenatal metabolism. Rock pigeons (Columba livia) typically lay 2-egg clutches with yolk androgen levels in second-laid eggs being consistently higher than in first-laid eggs. We investigated whether embryonic heart rate was higher in second- than first-laid eggs. Additionally, we increased yolk androgen levels (testosterone and androstenedione) with the mean difference between those in first- and second-laid eggs, to investigate whether the effects of androgens are egg sequence dependent. As expected, embryonic heart rate predicted body embryo organ- and body mass, and body dimensions, with body mass being significantly higher in second- than first-laid eggs. Androgen treated first-laid eggs increased heart rate to that of second-laid control eggs only temporally, yet it had an overall positive effect on embryo body dimensions but not on tissue mass. Our findings indicate that embryos from different egg laying sequence differed in heart rate and prenatal development outcomes but this can only partially be explained by their difference in maternal androgen levels.

Differential temperature effects on photoperiodism in female voles: A possible explanation for declines in vole populations.

Many mammalian species use photoperiod as a predictive cue to time seasonal reproduction. In addition, metabolic effects on the reproductive axis may also influence seasonal timing, especially in female small, short-lived mammals. To get a better understanding of how annual cycling environmental cues impact reproductive function and plasticity in small, short-lived herbivores with different geographic origins, we investigated the mechanisms underlying integration of temperature in the photoperiodic-axis regulating female reproduction in a Northern vole species (tundra vole, Microtus oeconomus) and in a Southern vole species (common vole, Microtus arvalis). We show that photoperiod and temperature interact to determine appropriate physiological responses; there is species-dependent annual variation in the sensitivity to temperature for reproductive organ development. In common voles, temperature can overrule photoperiodical spring-programmed responses, with reproductive organ mass being higher at 10°C than at 21°C, whereas in autumn they are less sensitive to temperature. These findings are in line with our census data, showing an earlier onset of spring reproduction in cold springs, while reproductive offset in autumn is synchronized to photoperiod. The reproductive organs of tundra voles were relatively insensitive to temperature, whereas hypothalamic gene expression was generally upregulated at 10°C. Thus, both vole species use photoperiod, whereas only common voles use temperature as a cue to control spring reproduction, which indicates species-specific reproductive strategies. Due to global warming, spring reproduction in common voles will be delayed, perhaps resulting in shorter breeding seasons and thus declining populations, as observed throughout Europe.

Is imitational learning a driving factor for the population bias in human hand preference?

Lateral preference is a widespread organizational principle in human and nonhuman animals. In humans, the most apparent lateralized trait (handedness) is unique in the animal kingdom because of a very pronounced bias towards right-handedness on a population level. In this study, based on previous experiments, we test the hypothesis that this bias was-among other factors-shaped by evolution through the facilitation of social learning. We exposed 134 subjects to footage of right- or left-handed knot making and analyzed whether concordant handedness between instructor and student facilitated quicker and more successful imitation. We used a set of nautical knots of different difficulty levels in order to test whether the potential effect of concordance became stronger with increasing knot difficulty. For all three performance measures (time until correct completion, number of attempts needed and correct imitation), we found hand congruency and difficulty level to be significant predictors but not the interaction of the two. We conclude that concordance of handedness between teacher and student of a motor skill enhances the speed and accuracy of imitation, which may have been a beneficial trait for selection to act upon, thereby shaping the human population bias in handedness.

The importance of understanding function and evolution.

Ocklenburg et al. (2020, Laterality 2020: Entering the next decade. Laterality) provided the field of laterality research with a stimulating research perspective for the coming decade, based on the current state of the art in both animal and human laterality research. Although this is paper takes many different approaches of laterality into account, we emphasize that the eco-evolutionary approach needs more attention. This concerns the question why organisms are lateralized in the first place, in other words, how does lateralization enhance the Darwinian fitness of the individual. We argue that laterality can be distinguished along four dimensions, and that each of them requires different ultimate explanations. Studying these functional and evolutionary explanations requires the development of ecologically relevant tests, adapted to the species at hand. It also requires experimental manipulation of laterality, testing the effect in (semi)-natural conditions on fitness parameters. Tools for such manipulation of laterality urgently require a better understanding of the developmental plasticity of lateralization, extending the field of evo-devo to that of eco-evo-devo. We also warn against seeing the minority in the distribution of direction or strength of lateralization as being a pathology as such minorities in biology can often be explained as adaptations by natural frequency dependent selection.

Practice of Noseband Use and Intentions Towards Behavioural Change in Dutch Equestrians.

Understanding equestrians' noseband tightening practices and intentions is necessary to target welfare improvement strategies. Firstly, we measured tightness in dressage and show jumping horses in The Netherlands, shortly after implementation of the two-finger rule by the Royal Dutch Equestrian Federation. Noseband tightness decreased with age, was less tight in dressage horses than in show jumpers, and was dependent on the interaction between competition level and discipline. Fifty-nine percent of the riders tightened nosebands to such an extent that they adhered to the new regulation. Secondly, we conducted an online survey to gain insight into whether riders were aware of noseband use and tightening behaviour. Of the 386 respondents, 54.5% agreed with the new regulations, and 62% believe that it improves horses' welfare. Applying cluster analysis to statements regarding their own attitude, peer pressure, and behavioural control produced three clusters. Noticeably, a lower percentage of Cluster 1 respondents (38%) performing at higher levels was convinced that the new regulation improved welfare than Cluster 2 (77.9%) and 3 (89.0%) respondents. Designing strategies to ensure the successful implementation of the new regulation and to convince equestrians to comply would be most effective if targeted differentially, and should include a transparent and objective form of regulation.

Does paternal immunocompetence affect offspring vulnerability to maternal androgens? A study in domestic chickens.

Exposure of yolk androgens can positively stimulate chick growth and competitive ability, but may negatively affect immunity. It has been hypothesized that only chicks from immunologically superior fathers can bear the cost of prenatal exposure to high androgen levels. To test this hypothesis, we paired roosters from two selection lines, one up- and one down-selected for natural antibodies (NAbs), with hens from a control line. We measured yolk testosterone and androstenedione levels, and we injected the treatment group of eggs of each female with testosterone suspended in sesame oil and the control group with sesame oil only. We then measured hatching success and growth, and characterized the humoral and cellular immune responses using three different challenges: a phyto-hemagglutinin, a lipopolysaccharide and a sheep red blood cell challenge. We found that the hatching success, body mass, initial levels of natural antibodies and the chicks' immunological responses to the three different challenges and development were affected neither by paternal immunocompetence nor by treatment. These results do not support the hypothesis that chicks from low-NAb line fathers are more sensitive to testosterone exposure during embryonic development than chicks from high-NAb line fathers.

The relationship between male social status, ejaculate and circulating testosterone concentration and female yolk androgen transfer in red junglefowl (Gallus gallus).

Several studies show that avian females prefer males based on their secondary sexual ornaments and dominance status. We tested in red junglefowl (Gallus gallus) how comb size affected the result of fighting and how the dominance status related to testosterone concentrations in their circulation and ejaculates. We subsequently tested how social status was related to female reproductive investment, including yolk hormone transfer. We found that after a fight 1) winners increased plasma T and decreased ejaculates T whereas losers' T remained unchanged, and 2) plasma T of winners was higher but ejaculates T was lower than those of losers. We argued those are consistent with the different reproductive strategies of dominant and subordinate males. Furthermore, in line with offspring sex-dependent growth patterns females transferred significantly more androstenedione to female than male embryos when mated with winners, while doing the opposite when mated with losers. We concluded therefore that female reproductive investment was affected by both partner quality and embryo sex. The results indicate that male quality influences sex-specific maternal investment, which could be mediated by ejaculate testosterone concentration.

Ejaculate testosterone levels affect maternal investment in red junglefowl (Gallus gallus gallus).

Substantial concentrations of testosterone are not only present in a male's circulation, but also in its ejaculate. Surprisingly, the regulation of ejaculate T and its effects on females and their offspring, potentially a cryptic paternal trait, are not known. We found lower circulating and higher ejaculate T concentrations in subordinate red junglefowl (Gallus gallus gallus) males compared to dominant males, suggestive of an adaptive trade-off in T allocation to circulation and their ejaculate. Subsequently, we artificially inseminated females with either testosterone enriched (TE) or control ejaculates (CE) in a cross-over design. TE females produced heavier eggs than CE females. Offspring growth and tonic immobility were affected in a sex-specific way by TE. TE sons were heavier with shorter TI duration than CE sons, and TE daughters were lighter than CE daughters but daughters did not differ in TI score. However, the chicks competitiveness was not influenced by the TE nor CE. This indicates a previously unknown function of ejaculate testosterone as well as a new form of interaction between a cryptic paternal trait and a maternal effect that may be widespread in the animal kingdom.

An adaptive annual rhythm in the sex of first pigeon eggs.

When the reproductive value of male and female offspring varies differentially, parents are predicted to adjust the sex ratio of their offspring to maximize their fitness (Trivers and Willard, Science 179:90-92, 1973). Two factors have been repeatedly linked to skews in avian offspring sex ratio. First, laying date can affect offspring sex ratio when the sexes differ in age of first reproduction, such that the more slowly maturing sex is overproduced early in the season. Second, position of the egg in the laying sequence of a clutch may affect sex ratio bias since manipulating the sex of the first eggs may be least costly to the mother. We studied both factors in two non-domesticated pigeon species. Both the Wood pigeon (Columba palumbus) and the Rock pigeon (Columba livia) have long breeding seasons and lay two-egg clutches. In the field, we determined the sex of Wood pigeon nestlings. In Rock pigeons, housed in captivity outdoors, we determined embryo sex after 3 days of incubation. On the basis of their sex-specific age of first reproduction, we predicted that males, maturing at older age than females, should be produced in majority early and females later in the year. This was confirmed for both species. The bias was restricted to first eggs. Rock pigeons produced clutches throughout the year and show that the sex of the first egg followed an annual cycle. To our knowledge, this study presents the first evidence of a full annual rhythm in adaptive sex allocation in birds. We suggest that this reflects an endogenous seasonal program in primary sex ratio controlled by a preovulatory mechanism.