When are egg-rejection cues perceived? A test using thermochromic eggs in an avian brood parasite host.

At the core of recognition systems research are questions regarding how and when fitness-relevant decisions made. Studying egg-rejection behavior by hosts to reduce the costs of avian brood parasitism has become a productive model to assess cognitive algorithms underlying fitness-relevant decisions. Most of these studies focus on how cues and contexts affect hosts' behavioral responses to foreign eggs; however, the timing of when the cues are perceived for egg-rejection decisions is less understood. Here, we focused the responses of American robins Turdus migratorius to model eggs painted with a thermochromic paint. This technique modified an egg's color with predictably varying temperatures across incubation: at the onset of incubation, the thermochromic model egg was cold and perceptually similar to a static blue model egg (mimicking the robin's own blue-green egg color), but by the end of an incubation bout, it was warm and similar to a static beige egg (mimicking the ground color of the egg of the robin's brood parasite, the brown-headed cowbird Molothrus ater). Thermochromic eggs were rejected at statistically intermediate rates between those of the static blue (mostly accepted) and static beige (mostly rejected) model eggs. This implies that at the population level, egg-rejection relevant cues are not perceived solely when arriving to or solely when departing from the nest. We also found that robins rejected their own eggs more often when exposed to color-changing model eggs relative to static eggs, suggesting that recognizing variable foreign eggs entails costly rejection errors for this host species.

Mimicry-dependent lateralization in the visual inspection of foreign eggs by American robins.

Brain lateralization, or the specialization of function in the left versus right brain hemispheres, has been found in a variety of lineages in contexts ranging from foraging to social and sexual behaviours, including the recognition of conspecific social partners. Here we studied whether the recognition and rejection of avian brood parasitic eggs, another context for species recognition, may also involve lateralized visual processing. We focused on American robins (Turdus migratorius), an egg-rejecter host to occasional brood parasitism by brown-headed cowbirds (Molothrus ater) and tested if robins preferentially used one visual hemifield over the other to inspect mimetic versus non-mimetic model eggs. At the population level, robins showed a significantly lateralized absolute eyedness index (EI) when viewing mimetic model eggs, but individuals varied in left versus right visual hemifield preference. By contrast, absolute EI was significantly lower when viewing non-mimetic eggs. We also found that robins with more lateralized eye usage rejected model eggs at higher rates. We suggest that the inspection and recognition of foreign eggs represent a specialized and lateralized context of species recognition in this and perhaps in other egg-rejecter hosts of brood parasites.

The chemical basis of a signal of individual identity: shell pigment concentrations track the unique appearance of Common Murre eggs.

In group-living species with parental care, the accurate recognition of one's own young is critical to fitness. Because discriminating offspring within a large colonial group may be challenging, progeny of colonial breeders often display familial or individual identity signals to elicit and receive parental provisions from their own parents. For instance, the common murre (or common guillemot: Uria aalge) is a colonially breeding seabird that does not build a nest and lays and incubates an egg with an individually unique appearance. How the shell's physical and chemical properties generate this individual variability in coloration and maculation has not been studied in detail. Here, we quantified two characteristics of the avian-visible appearance of murre eggshells collected from the wild: background coloration spectra and maculation density. As predicted by the individual identity hypothesis, there was no statistical relationship between avian-perceivable shell background coloration and maculation density within the same eggs. In turn, variation in both sets of traits was statistically related to some of their physico-chemical properties, including shell thickness and concentrations of the eggshell pigments biliverdin and protoporphyrin IX. These results illustrate how individually unique eggshell appearances, suitable for identity signalling, can be generated by a small number of structural mechanisms.

Probing the Limits of Egg Recognition Using Egg Rejection Experiments Along Phenotypic Gradients.

Brood parasites lay their eggs in other females' nests, leaving the host parents to hatch and rear their young. Studying how brood parasites manipulate hosts into raising their young and how hosts detect parasitism provide important insights in the field of coevolutionary biology. Brood parasites, such as cuckoos and cowbirds, gain an evolutionary advantage because they do not have to pay the costs of rearing their own young. However, these costs select for host defenses against all developmental stages of parasites, including eggs, their young, and adults. Egg rejection experiments are the most common method used to study host defenses. During these experiments, a researcher places an experimental egg in a host nest and monitors how hosts respond. Color is often manipulated, and the expectation is that the likelihood of egg discrimination and the degree of dissimilarity between the host and experimental egg are positively related. This paper serves as a guide for conducting egg rejection experiments from describing methods for creating consistent egg colors to analyzing the findings of such experiments. Special attention is given to a new method involving uniquely colored eggs along color gradients that has the potential to explore color biases in host recognition. Without standardization, it is not possible to compare findings between studies in a meaningful way; a standard protocol within this field will allow for increasingly accurate and comparable results for further experiments.

How to Make a Mimic? Brood Parasitic Striped Cuckoo Eggs Match Host Shell Color but Not Pigment Concentrations.

Hosts of avian brood parasites often use visual cues to reject foreign eggs, and several lineages of brood parasites have evolved mimetic eggshell coloration and patterning to circumvent host recognition. What is the mechanism of parasitic egg color mimicry at the chemical level? Mimetic egg coloration by Common Cuckoos Cuculus canorus is achieved by depositing similar concentrations of colorful pigments into their shells as their hosts. The mechanism of parasitic egg color mimicry at the chemical level in other lineages of brood parasites remains unexplored. Here we report on the chemical basis of egg color mimicry in an evolutionarily independent, and poorly studied, host-parasite system: the Neotropical Striped Cuckoo Tapera naevia and one of its hosts, the Rufous-and-white Wren Thryophilus rufalbus. In most of South America, Striped Cuckoos lay white eggs that are identical to those of local host species. In Central America, however, Striped Cuckoos lay blue eggs that match those of the Rufous-and-white Wren, suggesting that blue egg color in these cuckoo populations is an adaptation to mimic host egg appearance. Here we confirm that Striped Cuckoo eggs are spectrally similar to those of their hosts and consistently contain the same major eggshell pigment, biliverdin. However, wren eggshells lacked protoporphyrin, which was present in the parasitic cuckoo eggshells. Furthermore, biliverdin concentrations were significantly lower in cuckoo eggshells than in host eggshells. Similarity of host-parasite eggshell appearance, therefore, need not always be paralleled by a quantitative chemical match to generate effective visual mimicry in birds.

Eggshells as hosts of bacterial communities: An experimental test of the antimicrobial egg coloration hypothesis.

Oviparous animals have evolved multiple defenses to prevent microbes from penetrating their eggs and causing embryo mortality. In birds, egg constituents such as lysozyme and antibodies defend against microbial infestation, but eggshell pigments might also impact survival of bacteria. If so, microbes could exert an important selective pressure on the evolution of eggshell coloration. In a previous lab experiment, eggshell protoporphyrin caused drastic mortality in cultures of Gram positive, but not Gram negative, bacteria when exposed to light. Here, we test this "photodynamic antimicrobial hypothesis" in a field experiment. In a paired experimental design, we placed sanitized brown, protoporphyrin-rich chicken eggs alongside white eggs that lack protoporphyrin. We deployed eggs for 48 hr without incubation, as can occur between laying and incubation, when microbial infection risk is highest. Eggs were placed on the open ground exposed to sunlight and in dark underground storm-petrel burrows. We predicted that the proportion of Gram-positive bacteria on brown eggs should be lower when exposed to sunlight than when kept in the dark, but we expected no such difference for white eggs. Although our data revealed variation in bacterial community composition, the proportion of Gram-positive bacteria on eggshells did not vary by egg color, and there was no interaction between egg color and location. Instead, Gram-positive bacteria were proportionally more common on eggs on the ground than eggs in burrows. Overall, our experiment did not support the photodynamic antimicrobial hypothesis. The diverse range of avian egg colors is generated by just two pigments, but over 10 hypotheses have been proposed for the evolution of eggshell color. If our results are generalizable, eggshell protoporphyrin might not play a substantial role in defending eggs against microbes, which narrows the field of candidate hypotheses for the evolution of avian eggshell coloration.

Does contrast between eggshell ground and spot coloration affect egg rejection?

Obligate avian brood parasitic species impose the costs of incubating foreign eggs and raising young upon their unrelated hosts. The most common host defence is the rejection of parasitic eggs from the nest. Both egg colours and spot patterns influence egg rejection decisions in many host species, yet no studies have explicitly examined the role of variation in spot coloration. We studied the American robin Turdus migratorius, a blue-green unspotted egg-laying host of the brown-headed cowbird Molothrus ater, a brood parasite that lays non-mimetic spotted eggs. We examined host responses to model eggs with variable spot coloration against a constant robin-mimetic ground colour to identify patterns of rejection associated with perceived contrast between spot and ground colours. By using avian visual modelling, we found that robins were more likely to reject eggs whose spots had greater chromatic (hue) but not achromatic (brightness) contrast. Therefore, egg rejection decision rules in the American robin may depend on the colour contrast between parasite eggshell spot and host ground coloration. Our study also suggests that egg recognition in relation to spot coloration, like ground colour recognition, is tuned to the natural variation of avian eggshell spot colours but not to unnatural spot colours.