Control Broods Research Articles

Anthropogenic noise disrupts early-life development in a fish with paternal care

Anthropogenic noise is a global pollutant but its potential impacts on early life-stages in fishes are largely unknown. Here, using controlled laboratory experiments, we tested for impacts of continuous or intermittent exposure to low-frequency broadband noise on early life-stages of the common goby (Pomatoschistus microps), a marine fish with exclusive paternal care. Neither continuous nor intermittent noise exposure had an effect on filial cannibalism, showing that males were capable and willing to care for their broods. However, broods reared in continuous noise covered a smaller area and contained fewer eggs than control broods. Moreover, although developmental rate was the same in all treatments, larvae reared by males in continuous noise had, on average, a smaller yolk sac at hatching than those reared in the intermittent noise and control treatments, while larvae body length did not differ. Thus, it appears that the increased consumption of the yolk sac reserve was not utilised for increased growth. This suggests that exposure to noise in early life-stages affects fitness-related traits of surviving offspring, given the crucial importance of the yolk sac reserve during the early life of pelagic larvae. More broadly, our findings highlight the wide-ranging impacts of anthropogenic noise on aquatic wildlife living in an increasingly noisy world.

Hygienic behavioural response of honey bees (Apis mellifera and Apis cerana) against pin-killed brood as trait evaluation in Kashmir

In Apis mellifera and Apis cerana hygienic behavior involves recognition and removal of sick, damaged or dead brood from capped cells. We investigated whether bees react in the same way to grouped versus isolated damaged capped brood cells. Ten colonies of Apis mellifera honey bees and ten colonies of Apis cerana honey bees were used for this investigation. Capped worker brood cells aged 12 to 14 days old were perforated with the pin-killing method. After making holes in the brood cells, the combs were placed back into the hives; 24 h later the number of cleaned cells was recorded in areas with pin-killed and control brood cells. Four repetitions were made in each colony. Isolated cells were more frequently cleaned than grouped cells, though variance analysis showed no significant difference. Apis cerana bees also were somewhat, though not significantly more hygienic than Apis mellifera honey bees with mean removal of pin killed brood 97.46+/-1.37% and 83.83+/1.13% respectively. We conclude that honey bees can detect and remove both isolated and grouped dead brood. The tendency towards greater hygienic efficiency directed towards grouped Hygienic behaviour in honey bees was studied for first time with the aim of understanding mechanisms of pathogen resistance and colony health. It shows how honey bees confer colony level resistance and remove dead and diseased brood from the colony. This hygienic behavioural study can potentially increase the procedure of screening colonies for hygiene and breeding.

The interactive effect of ambient temperature and brood size manipulation on nestling body mass in blue tits: an exploratory analysis of a long-term study

BackgroundRelatively few studies have examined the interactive effects of ecological factors on physiological responses in wild animals. Nearly all of them have been short-term investigations that did not include experimental manipulations, limiting our ability to understand how climate change will affect natural populations. Using a 10-year brood size manipulation experiment in wild blue tits (Cyanistes caeruleus), we quantified the impact of weather conditions and brood competition on the body mass and structural size (tarsus length) of nestlings just prior to leaving the nest.ResultsWe found that variation in nestling body mass on day 14 after hatching was explained by an interactive effect between average ambient temperature experienced during nestling period and brood size treatment. Specifically, in control broods nestling body mass was correlated with temperature in a non-linear manner (concave) with the vertex point (maximum body mass) at ca. 13 °C. In contrast, in enlarged broods nestling body mass permanently increased (also non-linearly) as temperature advanced.ConclusionsOur results highlight the importance of considering the effects of brood rearing conditions alongside other environmental factors experienced during growth while investigating early-life environmental effects on body condition.

Nestling European Starlings (Sturnus vulgaris) adjust their begging calls in noise

ABSTRACT Anthropogenic noise, so common in cities, continues to increase with urbanisation. It adversely affects avian species that rely on acoustic forms of communication. The negative impacts are further exacerbated when parent-offspring communication is considered, especially in species where young are entirely dependent on the care of their parents. Our first objective was to study the effects that loud traffic noise had on nestling begging calls in European Starlings, Sturnus vulgaris, an urban-thriving species. For our second objective, we examined how this noise impacted parental provisioning and nestling condition. We found that the minimum frequency of the begging calls was higher in nestlings within experimental broods (exposed to traffic-noise playback) compared to that of nestlings in the control broods (exposed only to ambient noise). Also, nestlings in experimental broods continued to beg at a higher minimum frequency but with a narrowed bandwidth after the playback was stopped. Parental provisioning rates did not differ between control and experimental broods, nor did fledging success, although nestlings in the experimental group were in poorer condition. Our findings suggest that urban thrivers are affected by increasing traffic noise but have the phenotypic flexibility to adapt at a young age to maintain critical parent-offspring communication.

Epigenetics and Early Life Stress: Experimental Brood Size Affects DNA Methylation in Great Tits (Parus major)

Early developmental conditions are known to have life-long effects on an individual’s behavior, physiology and fitness. In altricial birds, a majority of these conditions, such as the number of siblings and the amount of food provisioned, are controlled by the parents. This opens up the potential for parents to adjust the behavior and physiology of their offspring according to local post-natal circumstances. However, the mechanisms underlying such intergenerational regulation remain largely unknown. A mechanism often proposed to possibly explain how parental effects mediate consistent phenotypic change is DNA methylation. To investigate whether early life effects on offspring phenotypes are mediated by DNA methylation, we cross-fostered great tit (Parus major) nestlings and manipulated their brood size in a natural study population. We assessed genome-wide DNA methylation levels of CpG sites in erythrocyte DNA, using Reduced Representation Bisulfite Sequencing (RRBS). By comparing DNA methylation levels between biological siblings raised in enlarged and reduced broods and between biological siblings of control broods, we assessed which CpG sites were differentially methylated due to brood size. We found 32 differentially methylated sites (DMS) between siblings from enlarged and reduced broods, a larger number than in the comparison between siblings from control broods. A considerable number of these DMS were located in or near genes involved in development, growth, metabolism, behavior and cognition. Since the biological functions of these genes line up with previously found effects of brood size and food availability, it is likely that the nestlings in the enlarged broods suffered from nutritional stress. We therefore conclude that early life stress might directly affect epigenetic regulation of genes related to early life conditions. Future studies should link such experimentally induced DNA methylation changes to expression of phenotypic traits and assess whether these effects affect parental fitness to determine if such changes are also adaptive.

Noise during mouthbrooding impairs maternal care behaviors and juvenile development and alters brain transcriptomes in the African cichlid fish Astatotilapia burtoni.

Anthropogenic noise has increased underwater ambient sound levels in the range in which most fishes detect and produce acoustic signals. Although the impacts of increased background noise on fish development have been studied in a variety of species, there is a paucity of information on how noise affects parental care. Mouthbrooding is an energetically costly form of parental care in which the brooding fish carries developing larvae in the buccal cavity for the duration of development. In the African cichlid Astatotilapia burtoni, females carry their brood for ~2 weeks during which time they do not eat. To test the hypothesis that increased background noise impacts maternal care behaviors and brood development, we exposed brooding females to a 3-h period of excess noise (~140 dB) played through an underwater speaker. Over half of noise-exposed brooding females cannibalized or pre-maturely released their brood, but 90% of control females exhibited normal brooding behaviors. RNA-seq analysis revealed that transcripts related to feeding and parental care were differentially expressed in the brains of noise-exposed females. Juveniles that were exposed to noise during their brood period within the mother's mouth had lower body condition factors, higher mortality and altered head transcriptomes compared with control broods. Furthermore, onset of adult-typical coloration and behaviors was delayed compared with control fish. Together, these data indicate that noise has severe impacts on reproductive fitness in mouthbrooding females. Our results, combined with past studies, indicate that parental care stages are extremely susceptible to noise-induced perturbations with detrimental effects on species persistence.

Extra food provisioning reduces extra‐pair paternity in the lesser kestrel Falco naumanni

Female promiscuity can function to acquire both direct and indirect benefits from their social mate and extra‐pair males. In many raptor species, intense mate‐feeding significantly contributes to female energy requirements before and during egg laying. Moreover, females may use mate‐feeding effort to assess male quality. In this study of the lesser kestrel Falco naumanni, we aimed at experimentally manipulating the female's perception of mate quality by providing females with extra food during egg laying, and evaluated the occurrence of extra‐pair paternity in food‐supplemented and control broods by parentage analyses. No extra‐pair offspring (EPO) was found among 19 food‐supplemented broods, whereas EPO occurred in five out of 17 control broods. No significant differences in morphological traits, body condition and reproductive success were found between faithful and unfaithful females. However, clutches containing EPO were laid later in the breeding season. Moreover, un‐cuckolded males had longer tarsi than cuckolded ones, indicating larger body size. Hence, extra food provisioning and early breeding reduced the occurrence of EPO in lesser kestrels. In addition, we confirmed the occurrence of intraspecific brood parasitism, as five nestlings were not the offspring of the brooding female. The results of our food‐provisioning experiment support the idea that mate‐feeding ability is a reliable indicator of male quality, and are in accordance with the hypothesis that male mate‐feeding behaviour is a sexually selected trait.

Physiological responses of host parents to rearing an avian brood parasite: An experimental study

Raising an obligate avian brood parasite is costly for host parents because it redirects valuable parental resources from one's own offspring to genetically unrelated young. The costs of raising a brood parasite may be mediated by physiological stressors for foster parents if it requires greater or biased parental effort compared to raising non-parasitized broods. For example, upregulating glucocorticoid hormones or reducing immune responses may mediate a trade-off between resource allocation to a current brood versus future reproductive opportunities, but published data on parasitized hosts' physiology are scarce. Here we used an experimental approach to test if host parents respond to the presence of brood parasitic young through dynamic changes in their own physiology. We conducted our experiments with parasitic brown-headed cowbirds (Molothrus ater) fostered into nests of host prothonotary warblers (Protonotaria citrea). Relative to parents caring for non-parasitized control broods, parasitism increased baseline corticosterone levels and reduced body mass in adult male, but not in female, hosts. Immune responses to a novel antigen were depressed in both parents of parasitized broods compared to parents of non-parasitized broods. Additionally, we found that immune function increased along the breeding season regardless of treatment. These experiments also confirmed prior observational data that parasitized adult males have reduced return rates to breeding sites in years subsequent to raising cowbirds. The findings demonstrate diverse physiological effects of parasitism on the foster parents in our particular host-brood parasite system, yet we found no evidence of individual trade-offs between endocrine and immune responses of adult hosts.

Efficacy of Erymicin 200 Injections for Reducing Renibacterium salmoninarum and Controlling Vertical Transmission in an Inland Rainbow Trout Brood Stock.

Bacterial Kidney Disease, caused by Renibacterium salmoninarum (Rs), is widespread and can cause significant mortality at most life stages in infected salmonids. Rs is commonly found in inland trout, which can be carriers of the bacterium. Lethal spawns can be used to control vertical transmission to progeny through the culling of eggs from infected parents, but can be costly, time-consuming, and can negatively impact important and rare brood stocks. Erymicin 200 is an Investigational New Animal Drug (INAD) intended to reduce Rs levels in hatchery brood stocks and control vertical transmission to progeny. We tested the efficacy of Erymicin 200 injections in a positive, hatchery-resident rainbow trout (Oncorhynchus mykiss) brood stock in Colorado, USA. Brood fish age two and three were injected with 25 mg per kg of body weight Erymicin 200 three times prior to spawning. Erymicin 200 was effective in reducing Rs to below detectable levels in treated fish. However, both negative treated and control brood fish produced positive progeny, suggesting that Erymicin 200 did not prevent the vertical transmission of Rs.

Providing urban birds nutritious food to feed chicks reduces urban versus rural breeding success disparities.

In Focus: Seress, G, Sándor, K, Evans, KL, Liker, A. (2020) Food availability limits avian reproduction in the city: An experimental study on great tits Parus major . Journal of Animal Ecology. 89. Animals in urban areas face a multitude of stressors, but how each stressor impacts survival and fitness can be difficult to disentangle. We need experimental manipulations of suspected stressors to examine causal relationships with traits of free-living urban and rural animals. In this issue, Seress and colleagues take on an intensive experimental approach to test whether one potential stressor-limited natural food sources in cities-can explain reduced avian reproductive success in urban areas. They employ a full factorial design, including both food supplemented and control broods in both urban and forest great tit Parus major populations. The findings are clear. Reduced food availability is the key factor limiting urban offspring growth and survival, at least in this well-studied species. Indeed, the extra insects fed to urban chicks greatly reduced the significant differences in survival rates and body sizes between urban and forest broods. The findings are also sobering. Urban insect populations would need to more than double to erase differences in reproductive success between urban and rural bird populations, an unlikely outcome with the ever-increasing urbanization of habitats.

Food availability limits avian reproduction in the city: An experimental study on great tits Parus major.

The altered ecological and environmental conditions in towns and cities strongly affect demographic traits of urban animal populations, for example avian reproductive success is often reduced. Previous work suggests that this is partly driven by low insect availability during the breeding season, but robust experimental evidence that supports this food limitation hypothesis is not yet available. We tested core predictions of the food limitation hypothesis using a controlled experiment that provided supplementary insect food (nutritionally enhanced mealworms supplied daily to meet 40%-50% of each supplemented brood's food requirements) to great tit nestlings in urban and forest habitats. We measured parental provisioning rates and estimated the amount of supplementary food consumed by control and experimental nestlings, and assessed their body size and survival rates. Provisioning rates were similar across habitats and control and supplemented broods, but supplemented (and not control) broods consumed large quantities of supplementary food. As predicted by the food limitation hypothesis we found that nestlings in (a) urban control broods had smaller body size and nestling survival rates than those in forest control broods; (b) forest supplemented and control broods had similar body size and survival rates; (c) urban supplemented nestlings had larger body size and survival rates than those in urban control broods; and crucially (d) urban supplemented broods had similar body size and survival rates to nestlings in forest control broods. Our results provide rare experimental support for the strong negative effects of food limitation during the nestling rearing period on urban birds' breeding success. Furthermore, the fact that supplementary food almost completely eliminated habitat differences in survival rate and nestling body size suggest that urban stressors other than food shortage contributed relatively little to the reduced avian breeding success. Finally, given the impacts of the amount of supplementary food that we provided and taking clutch size differences into account, our results suggest that urban insect populations in our study system would need to be increased by a factor of at least 2.5 for urban and forest great tits to have similar reproductive success.

Effect of food availability on offspring sex ratios in replacement clutches of Mew Gulls (Larus canus) and Black-headed Gulls (Chroicocephalus ridibundus) in the highly unstable environment of the Vistula River

The reproductive cost hypothesis predicts that parents with low investment capacity (e.g., those in poor condition) should produce the larger sex (usually males) less often so as to minimize the risk of current reproductive failure and/or a decrease in their residual reproductive value. The study aimed to test whether offspring sex ratio bias in replacement clutches of the Mew Gull (MG; Larus canus) and Black-headed Gull (BHG; Chroicocephalus ridibundus) exposed to different food conditions was in agreement with predictions of the reproductive cost hypothesis. Food availability was experimentally modified before egg laying. Our experimental treatment affected hatching sex ratio, being also significant the effect of laying order and species identity. For both gull species, parents that were provided with supplemental food had more sons at hatching than unfed (control) pairs. This was particularly so if they were fed until production of a replacement clutch. This gradient in hatching sex ratio was much more pronounced in MG than in BHG. In both gull species, sons hatched more often from first laid (A) than from second (B) or third-laid egg (C). Five days after hatching, the sex ratio of surviving chicks was dependent mainly on the experimental treatment and egg sequence. Unlike the time of hatching, there was no important effect of species × treatment interaction. In both gull species, the brood sex-ratio was clearly male-biased in parents receiving supplemental feeding. However, differences between experimental groups were less extreme than at hatching. Chicks hatched from C-egg had much lower survival than chicks from the first two eggs (A- and B-egg), irrespective of sex, treatment, or species. Supplementary feeding was associated with improved chick survival for both gull species, although this effect was dependent on chick gender. Surprisingly, in the case of control broods (i.e., broods whose parents did not receive supplementary food), male chicks survived better than female chicks, but the opposite was true for broods in which parents were fed. There was an overall tendency for higher survival of BHG chicks than MG chicks. During the early-chick period, competition between siblings and the related differentiated mortality of chicks from A-, B-, and C-eggs, constituted a mechanism which shaped the brood sex ratio bias. In this context, differences in the sex ratio bias in eggs of different rank (A, B, C) at hatching were important. The results appear to fulfil the expectations of the reproductive cost hypothesis. For the species with greater sexual dimorphism (MG), a relationship between the condition of parents and the expected bias of sex ratio was visible only at hatching, which may indicate that this feature is not significant in shaping the sex ratio bias in MG.

Benefits of an anti-parasite treatment are influenced by within-brood size variation in Tree Swallows (Tachycineta bicolor)

AbstractIn all animals, susceptibility to parasites can differ among individuals. Young, nest-bound birds are exposed to a diversity of nest-dwelling ectoparasites that typically feed on their blood. Within broods, hatching asynchrony creates size hierarchies that result in morphological and physiological variation among nest mates, and susceptibility to parasites also may vary predictably with this size hierarchy. Our objective was to use a broad-spectrum, anti-parasite drug, ivermectin (IVM), to treat individual nestling Tree Swallows (Tachycineta bicolor) and assess how nestling susceptibility to parasites varied both within and among broods. Broods were either assigned to an IVM group, where half of the nestlings in a brood received IVM injections and half received control injections of pure sesame oil, or to a control group, where all nestlings received oil injections. We found that the IVM treatment reduced parasite loads for broods as a whole, thereby benefiting all nestlings in IVM broods and suggesting our treatment resulted in herd immunity. Specifically, nestlings from IVM broods had higher hemoglobin concentrations, regardless of whether they received injections with IVM or oil, and greater fledging success, than nestlings from control broods. On the contrary, IVM treatment did not strongly affect nestling morphology, with only marginal effects on the growth rate of ninth primary feathers, and the effects of the treatment on 2 other morphological traits depending on temporal factors. Variation in size within broods, however, influenced the chance of an individual fledging, which increased with relative size within a brood, but only under lower parasite loads (i.e. IVM broods). By experimentally manipulating nestling susceptibility to parasites, we have demonstrated variation in nestling response to an anti-parasite treatment both within and among broods, and future studies should investigate the underlying mechanism for why certain nestlings along the brood size hierarchy are more susceptible to parasites.

Exposure to predator models during the fertile period leads to higher levels of extra-pair paternity in blue tits.

The perceived risk of predation can affect breeding behaviour and reduce reproductive success in prey species. Individuals exposed to predators may also adopt different mating tactics with potential consequences for the distribution of paternity in socially monogamous species that engage in extra-pair copulations. We experimentally increased perceived predation risk during the fertile period in blue tits Cyanistes caeruleus. Every morning between nest completion and the onset of egg laying, we presented a model of either a predator or a non-predator (control) near active nestboxes. Broods from pairs exposed to predators had higher levels of extra-pair paternity than control broods. This mainly resulted from a higher proportion of extra-pair offspring in broods with at least one extra-pair young. Females exposed to predators first emerged from the nestbox later in the morning, stayed away from the nestbox for longer and were less likely to be visited at the nest by their social mate, but we detected no behavioural differences once the model was removed. Our results suggest that the higher rates of extra-pair paternity resulted from the disruption of morning routines, which may have inhibited within-pair copulations or increased opportunities for females to engage in extra-pair copulations. We conclude that the perceived risk of predation can have substantial effects on levels of extra-pair paternity.

Experimentally flight-impaired females show higher levels of extra-pair paternity in the pied flycatcher Ficedula hypoleuca.

There is no consensus yet on the reasons why females engage in extra-pair copulations (EPCs). In some species, females have been shown to accrue some indirect benefits, but these effects are not consistent across species and studies. The sexual conflict hypothesis posits that extra-pair paternity (EPP) is the result of strong selection for male pursuit of EPC without real benefits for females. In order to test this hypothesis, we experimentally reduced wing area (reversibly tying together some primary feathers), in a group of pied flycatcher females (Ficedula hypoleuca). The manipulation increases wing loading (body mass/wing area), which is negatively associated with flying ability, and thus with the capacity to escape from unwanted copulations. We compared the levels of EPP in this experimental group with those of a group of un-manipulated females. Experimental females almost doubled the proportion of extra-pair young (EPY) with respect to control females. In addition, more males sired EPY in experimental than in control broods containing EPY. These results suggest that in our study population, EPP could be partially a product of female capacity to avoid EPCs. We also discuss the alternative hypothesis that results might be due to an eventual reduction of female attractiveness.

Bird embryos perceive vibratory cues of predation risk from clutch mates.

During development in fluctuating environments, phenotypes can be adjusted to the conditions that individuals will probably encounter later in life. As developing embryos have a limited capacity to fully capture environmental information, theory predicts that they should integrate relevant information from all reliable sources, including the social environment. In many oviparous species, embryos are able to perceive cues of predator presence in some circumstances, but whether this information is socially transmitted among clutch mates-promoting phenotypic adjustments in the whole clutch-is unknown. Here, using an experimental design for which we modified the exposure to some, but not all, embryos of the same clutch to cues of predator presence (that is, alarm calls), we show that exposed embryos of the yellow-legged gull (Larus michahellis) and their unexposed clutch mates showed similar developmental changes that were absent in embryos from control clutches. Compared with the control broods, both embryos that were exposed to alarm calls and their unexposed clutch mates showed altered prenatal and postnatal behaviours, higher levels of DNA methylation and stress hormones, and reduced growth and numbers of mitochondria (which may be indicative of the capacity for energy production of cells). These results strongly suggest that gull embryos are able to acquire relevant environmental information from their siblings. Together, our results highlight the importance of socially acquired information during the prenatal stage as a non-genetic mechanism promoting developmental plasticity.

Stock-specific chemical brood signals are induced by Varroa and Deformed Wing Virus, and elicit hygienic response in the honey bee

The health of the honey bee Apis mellifera is challenged by the ectoparasitic mite Varroa destructor, and the numerous harmful pathogens it vectors. Existing pesticide-based Varroa controls are not sustainable. In contrast, one promising approach for improved honey bee health is the breeding of hygienic bees, capable of detecting and removing brood that is parasitized or diseased. In three experiments we find evidence to support the hypothesis that stock-specific chemical brood signals are induced by Varroa and Deformed Wing Virus, and elicit hygienic response in the honey bee. By collecting, analyzing, and running bioassays involving mite-infested and control brood extracts from three honey bee breeding stocks we: 1) found evidence that a transferrable chemical signal for hygienic behavior is present in Varroa-infested brood extracts, 2) identified ten stock-specific hydrocarbons as candidates of hygienic signaling, and 3) found that two of these hydrocarbons linked to Varroa and DWV were also elevated in brood targeted for hygienic behavior. These findings expand our understanding of honey bee chemical communication, and facilitate the development of improved hygienic selection tools to breed honey bees with greater resistance to Varroa and associated pathogens.

Colony size affects nestling immune function: a cross-fostering experiment in a colonial waterbird

Elevated transmission rate of pathogens and parasites is considered one of the major costs of sociality in birds. However, greater risk of infection in colonial birds might be compensated by specific immune adaptations. Here, we predicted that nestlings raised in larger colonies should invest more in their immune function. To test this hypothesis, we manipulated colony size and conduced cross-fostering experiment in a colonial waterbird, the common tern Sterna hirundo. Establishment of different size colonies under uniform environmental conditions was induced by providing large and small patches of attractive nesting area for terns (floating rafts). Then, pairs of clutches were swapped between large and small tern colonies, and skin-swelling response to phytohaemagglutinin (PHA) was assessed for nestlings from experimental and control broods. Contrary to our expectations, we found a negative effect of foster colony size on nestling PHA response (nestlings raised in the larger colony had lower PHA response). In addition, nestling PHA response correlated negatively with heterophil/lymphocyte ratio used as a measure of physiological stress. This suggested that low PHA response of nestlings raised in the larger colony could be mediated by an elevated level of social stress. We suggest that depression of immune function via social stress may constitute a strong selective pressure against large colony size in the common tern, and possibly in other colonial species. We also recommend that this largely overlooked cost of sociality should be considered in the further studies on the evolution and ecology of avian coloniality.

Maternal investment in last-laid eggs does not compensate for hatching asynchrony in a seabird.

Maternal and environmental effects can have profound effects on offspring performance by generating variation in offspring phenotypes, independent of genetic effects. Within avian broods, differential maternal investment of resources across the laying sequence is thought to be an adaptive strategy to modulate competitive hierarchies induced by hatching asynchrony. In this study, we evaluated the relative importance of maternally derived within-clutch variation and the asymmetric post-hatching environment for growth and survival of common tern (Sterna hirundo) siblings. We experimentally manipulated hatching order, resulting in chicks from last-laid eggs hatching first and vice versa. Although both initial age and size asymmetries were larger within experimental than control broods, the early survival of last-hatched chicks was similar between groups. Initial positive effects of egg size disappeared as siblings approached fledging. Ultimately at fledging, both within-brood growth and cumulative survival patterns were similar between experimental and control broods, suggesting that the effects of systematic variation of egg constituents (e.g., maternally derived yolk hormones) and egg size are too subtle and largely overwhelmed by the effects of hatching asynchrony. Therefore, we conclude that variation in offspring phenotypes is pre-dominantly determined by the social environment experienced post-hatching. Maternal effects may further fine-tune phenotypic variation in response to varying environmental conditions, but this needs to be tested through empirical studies in which multiple maternal effects are measured simultaneously under different environmental conditions.

Brood size influences patterns of DNA methylation in wild Zebra Finches (Taeniopygia guttata)

Both natural and experimentally manipulated brood size can influence the competition dynamics among siblings in a nest and alter the environment during early development in birds. Brood size affects a variety of life-history and fitness-related traits, but relatively little is known about the mechanisms that might mediate these effects. There is accumulating evidence that early-life environments can influence adult phenotypes though epigenetic mechanisms such as variation in DNA methylation. Here, we profile DNA methylation in nestling Zebra Finches (Taeniopygia guttata) raised in naturally variable and experimentally manipulated brood sizes. We found that (1) natal brood size is significantly, positively correlated with percent DNA methylation in blood; and (2) individuals in manipulated broods experience significantly more demethylation events across early development than individuals from broods that remain the same size. Any manipulation of brood size creates fluctuations in early developmental conditions, potentially explaining why the frequency of demethylation events in these treatments was higher than in control broods. We also found that the specific loci that lost and gained methylation across early development differed between individuals in enlarged and reduced broods, which may reflect the different developmental pressures imposed by the different manipulations. Although the phenotypic consequences of reduced levels of methylation are yet to be elucidated, our findings support the hypothesis that brood size is associated with the prevalence and pattern of DNA methylation in wild birds.