Urbanization correlates with the prevalence and richness of blood parasites in Eurasian Blackbirds (Turdus merula).

Urbanization is increasing worldwide, producing severe environmental impacts. Biodiversity is affected by the expansion of cities, with many species being unable to cope with the different human-induced stressors present in these landscapes. However, this knowledge is mainly based on research from taxa such as plants or vertebrates, while other organisms like protozoa have been less studied in this context. The impact of urbanization on the transmission of vector-borne pathogens in wildlife is still unclear despite its relevance for animal and human health. Here, we investigated whether cities are associated with changes in the prevalence and richness of lineages of three vector-borne protozoans (Plasmodium, Haemoproteus and Leucocytozoon) in Eurasian blackbirds (Turdus merula) from multiple urban and forest areas in Europe. Our results show important species-specific differences between these two habitat types. We found a significant lower prevalence of Leucocytozoon in urban birds compared to forest birds, but no differences for Plasmodium and Haemoproteus. Furthermore, the richness of parasite lineages in European cities was higher for Plasmodium but lower for Leucocytozoon than in forests. We also found one Plasmodium lineage exclusively from cities while another of Leucocytozoon was only found in forests suggesting a certain level of habitat specialization for these protozoan vectors. Overall, our findings show that cities provide contrasting opportunities for the transmission of different vector-borne pathogens and generate new scenarios for the interactions between hosts, vectors and parasites.

Dense city centers support less evolutionary unique bird communities than sparser urban areas.

Urbanization alters avian communities, generally lowering the number of species and contemporaneously increasing their functional relatedness, leading to biotic homogenization. Urbanization can also negatively affect the phylogenetic diversity of species assemblages, potentially decreasing their evolutionary distinctiveness. We compare species assemblages in a gradient of building density in seventeen European cities to test whether the evolutionary distinctiveness of communities is shaped by the degree of urbanization. We found a significant decline in the evolutionary uniqueness of avian communities in highly dense urban areas, compared to low and medium-dense areas. Overall, communities from dense city centers supported one million years of evolutionary history less than communities from low-dense urban areas. Such evolutionary homogenization was due to a filtering process of the most evolutionarily unique birds. Metrics related to evolutionary uniqueness have to play a role when assessing the effects of urbanization and can be used to identify local conservation priorities.

Spatial Overlap and Habitat Selection of Corvid Species in European Cities.

Understanding habitat and spatial overlap in sympatric species of urban areas would aid in predicting species and community modifications in response to global change. Habitat overlap has been widely investigated for specialist species but neglected for generalists living in urban settings. Many corvid species are generalists and are adapted to urban areas. This work aimed to determine the urban habitat requirements and spatial overlap of five corvid species in sixteen European cities during the breeding season. All five studied corvid species had high overlap in their habitat selection while still having particular tendencies. We found three species, the Carrion/Hooded Crow, Rook, and Eurasian Magpie, selected open habitats. The Western Jackdaw avoided areas with bare soil cover, and the Eurasian Jay chose more forested areas. The species with similar habitat selection also had congruent spatial distributions. Our results indicate that although the corvids had some tendencies regarding habitat selection, as generalists, they still tolerated a wide range of urban habitats, which resulted in high overlap in their habitat niches and spatial distributions.

Effects of light and noise pollution on avian communities of European cities are correlated with the species' diet.

Urbanization affects avian community composition in European cities, increasing biotic homogenization. Anthropic pollution (such as light at night and noise) is among the most important drivers shaping bird use in urban areas, where bird species are mainly attracted by urban greenery. In this study, we collected data on 127 breeding bird species at 1349 point counts distributed along a gradient of urbanization in fourteen different European cities. The main aim was to explore the effects of anthropic pollution and city characteristics, on shaping the avian communities, regarding species' diet composition. The green cover of urban areas increased the number of insectivorous and omnivorous bird species, while slightly decreasing the overall diet heterogeneity of the avian communities. The green heterogeneity-a measure of evenness considering the relative coverage of grass, shrubs and trees-was positively correlated with the richness of granivorous, insectivorous, and omnivorous species, increasing the level of diet heterogeneity in the assemblages. Additionally, the effects of light pollution on avian communities were associated with the species' diet. Overall, light pollution negatively affected insectivorous and omnivorous bird species while not affecting granivorous species. The noise pollution, in contrast, was not significantly associated with changes in species assemblages. Our results offer some tips to urban planners, managers, and ecologists, in the challenge of producing more eco-friendly cities for the future.

Cloacal microbiota are biogeographically structured in larks from desert, tropical and temperate areas.

BACKGROUND: In contrast with macroorganisms, that show well-documented biogeographical patterns in distribution associated with local adaptation of physiology, behavior and life history, strong biogeographical patterns have not been found for microorganisms, raising questions about what determines their biogeography. Thus far, large-scale biogeographical studies have focused on free-living microbes, paying little attention to host-associated microbes, which play essential roles in physiology, behavior and life history of their hosts. Investigating cloacal gut microbiota of closely-related, ecologically similar free-living songbird species (Alaudidae, larks) inhabiting desert, temperate and tropical regions, we explored influences of geographical location and host species on α-diversity, co-occurrence of amplicon sequence variants (ASVs) and genera, differentially abundant and dominant bacterial taxa, and community composition. We also investigated how geographical distance explained differences in gut microbial community composition among larks. RESULTS: Geographic location did not explain variation in richness and Shannon diversity of cloacal microbiota in larks. Out of 3798 ASVs and 799 bacterial genera identified, 17 ASVs (< 0.5%) and 43 genera (5%) were shared by larks from all locations. Desert larks held fewer unique ASVs (25%) than temperate zone (31%) and tropical larks (34%). Five out of 33 detected bacterial phyla dominated lark cloacal gut microbiomes. In tropical larks three bacterial classes were overrepresented. Highlighting the distinctiveness of desert lark microbiota, the relative abundances of 52 ASVs differed among locations, which classified within three dominant and 11 low-abundance phyla. Clear and significant phylogenetic clustering in cloacal microbiota community composition (unweighted UniFrac) showed segregation with geography and host species, where microbiota of desert larks were distinct from those of tropical and temperate regions. Geographic distance was nonlinearly associated with pairwise unweighted UniFrac distances. CONCLUSIONS: We conclude that host-associated microbiota are geographically structured in a group of widespread but closely-related bird species, following large-scale macro-ecological patterns and contrasting with previous findings for free-living microbes. Future work should further explore if and to what extent geographic variation in host-associated microbiota can be explained as result of co-evolution between gut microbes and host adaptive traits, and if and how acquisition from the environmental pool of bacteria contributes to explaining host-associated communities.

EVI and NDVI as proxies for multifaceted avian diversity in urban areas.

Most ecological studies use remote sensing to analyze broad-scale biodiversity patterns, focusing mainly on taxonomic diversity in natural landscapes. One of the most important effects of high levels of urbanization is species loss (i.e., biotic homogenization). Therefore, cost-effective and more efficient methods to monitor biological communities' distribution are essential. This study explores whether the Enhanced Vegetation Index (EVI) and the Normalized Difference Vegetation Index (NDVI) can predict multifaceted avian diversity, urban tolerance, and specialization in urban landscapes. We sampled bird communities among 15 European cities and extracted Landsat 30-meter resolution EVI and NDVI values of the pixels within a 50-m buffer of bird sample points using Google Earth Engine (32-day Landsat 8 Collection Tier 1). Mixed models were used to find the best associations of EVI and NDVI, predicting multiple avian diversity facets: Taxonomic diversity, functional diversity, phylogenetic diversity, specialization levels, and urban tolerance. A total of 113 bird species across 15 cities from 10 different European countries were detected. EVI mean was the best predictor for foraging substrate specialization. NDVI mean was the best predictor for most avian diversity facets: taxonomic diversity, functional richness and evenness, phylogenetic diversity, phylogenetic species variability, community evolutionary distinctiveness, urban tolerance, diet foraging behavior, and habitat richness specialists. Finally, EVI and NDVI standard deviation were not the best predictors for any avian diversity facets studied. Our findings expand previous knowledge about EVI and NDVI as surrogates of avian diversity at a continental scale. Considering the European Commission's proposal for a Nature Restoration Law calling for expanding green urban space areas by 2050, we propose NDVI as a proxy of multiple facets of avian diversity to efficiently monitor bird community responses to land use changes in the cities.

Functional explanation of extreme hatching asynchrony: Male Manipulation Hypothesis.

Hatching asynchrony in birds is considered an adaptation to facilitate brood reduction because under conditions of food scarcity, the smallest nestling usually dies soon after hatching, thereby minimizing parental effort. However, in species with extreme hatching asynchrony, the last hatchlings paradoxically experience a very low probability of survival and death can take so long that it can hardly be considered an adaptation. Here, we propose and experimentally tested a new adaptive hypothesis explaining the brood reduction paradox, namely the "Male Manipulation Hypothesis". Our hypothesis suggests that by inducing asynchronous hatching, females increase the feeding requirements of the brood, which will induce males to increase provisioning effort. In addition, females may extend the period of male manipulation by feeding the smallest nestling just enough to sustain life. Our study showed that male common blackbirds ( Turdus merula) increased their effort (i.e., number of food items per hour) in experimental asynchronous broods compared to synchronous broods, while females reduced their contribution, as predicted by the hypothesis.

The extended avian urban phenotype: anthropogenic solid waste pollution, nest design, and fitness.

Solid waste pollution (garbage discarded by humans, such as plastic, metal, paper) has received increased attention given its importance as a global threat to biodiversity. Recent studies highlight how animals incorporate anthropogenic materials into their life-cycle, for example in avian nest construction. While increasingly monitored in natural areas, the influence of solid waste pollution on wildlife has been seldom explored in the urban habitat. There is limited data on the relationship between anthropogenic solid waste pollution, nest design, and reproductive success in an urban context. We address this knowledge gap (i) by investigating the presence of environmental solid waste pollution in the breeding habitats of great tits Parus major and blue tits Cyanistes caeruleus reproducing in a gradient of urbanisation, and (ii) by quantifying (ii) the contribution of different anthropogenic materials in their nests. We further examine potential drivers of solid waste pollution by inferring three distinct properties of the urban space: environmental solid waste pollution on the ground, human presence, and the intensity of urbanisation (e.g impervious surfaces) in nestbox vicinity. Finally, (iii) we explore the relationship between anthropogenic nest materials and reproductive success. We found that environmental solid waste pollution was positively associated with human presence and urbanisation intensity. There was also a positive relationship between increased human presence and the amount of anthropogenic materials in great tit nests. Interestingly, in both species, anthropogenic nest materials covaried negatively with nest materials of animal origin (fur and feathers). We suggest that fur and feathers - key insulating materials in nest design - may be scarcer in areas with high levels of human presence, and are consequently replaced with anthropogenic nest materials. Finally, we report a negative relationship between anthropogenic nest materials and blue tit reproductive success, suggesting species-specific vulnerability of urban birds to solid waste pollution.

Prolactin mediates behavioural rejection responses to avian brood parasitism.

Adaptations resulting from co-evolutionary interactions between avian brood parasites and their hosts have been extensively studied, yet the physiological mechanisms underlying antiparasitic host defences remain little known. Prolactin, one of the main hormones involved in the regulation of avian parental behaviour, might play a key role in the orchestration of the host responses to avian brood parasitism. Given the positive association between prolactin and parental behaviour during incubation, decreasing prolactin levels are expected to facilitate egg-rejection decisions. We tested this prediction by implanting Eurasian blackbird (Turdus merula) females with an inhibitor of prolactin secretion, bromocriptine mesylate, to experimentally decrease their plasma prolactin levels. Bromocriptine mesylate-implanted individuals ejected mimetic model eggs at higher rates, and showed shorter latency to egg ejection, than placebo-treated birds. To our knowledge, this is the first experimental evidence that behavioural host defences against avian brood parasitism are mediated by prolactin.

Effects of urbanization on taxonomic, functional and phylogenetic avian diversity in Europe.

Europe is an urbanized continent characterized by a long history of human-wildlife interactions. This study aimed to assess the effects of specific elements of urbanization and urban pollution on complementary avian diversity metrics, to provide new insights on the conservation of urban birds. Our study recorded 133 bird species at 1624 point counts uniformly distributed in seventeen different European cities. Our results thus covered a large spatial scale, confirming both effects of geographical and local attributes of the cities on avian diversity. However, we found contrasting effects for the different diversity components analyzed. Overall, taxonomic diversity (bird species richness), phylogenetic diversity and relatedness were significantly and negatively associated with latitude, while functional dispersion of communities showed no association whatsoever. At the local level (within the city), we found that urban greenery (grass, bush, and trees) is positively correlated with the number of breeding bird species, while the building cover showed a detrimental effect. Functional dispersion was the less affected diversity metric, while grass and trees and water (rivers or urban streams) positively affected the phylogenetic diversity of avian communities. Finally, the phylogenetic relatedness of species increased with all the main indicators of urbanization (building surface, floors, pedestrian's density and level of light pollution) and was only mitigated by the presence of bushes. We argue that maintaining adequate levels of avian diversity within the urban settlements can help to increase the potential resilience of urban ecosystems exposed to the stress provoked by rapid and continuous changes. We listed some characteristics of the cities providing positive and negative effects on each facet of urban avian diversity.

Physiological stress does not increase with urbanization in European blackbirds: Evidence from hormonal, immunological and cellular indicators.

Urbanization changes the landscape structure and ecological processes of natural habitats. While urban areas expose animal communities to novel challenges, they may also provide more stable environments in which environmental fluctuations are buffered. Species´ ecology and physiology may determine their capacity to cope with the city life. However, the physiological mechanisms underlying organismal responses to urbanization, and whether different physiological systems are equally affected by urban environments remain poorly understood. This severely limits our capacity to predict the impact of anthropogenic habitats on wild populations. In this study, we measured indicators of physiological stress at the endocrine, immune and cellular level (feather corticosterone levels, heterophil to lymphocyte ratio, and heat-shock proteins) in urban and non-urban European blackbirds (Turdus merula) across 10 European populations. Among the three variables, we found consistent differences in feather corticosterone, which was higher in non-urban habitats. This effect seems to be dependent on sex, being greater in males. In contrast, we found no significant differences between urban and non-urban habitats in the two other physiological indicators. The discrepancy between these different measurements of physiological stress highlights the importance of including multiple physiological variables to understand the impact of urbanization on species' physiology. Overall, our findings suggest that adult European blackbirds living in urban and non-urban habitats do not differ in terms of physiological stress at an organismal level. Furthermore, we found large differences among populations on the strength and direction of the urbanization effect, which illustrates the relevance of spatial replication when investigating urban-induced physiological responses.

Biodiversity within the city: Effects of land sharing and land sparing urban development on avian diversity.

Urbanization, one of the most extreme human-induced environmental changes, is negatively affecting biodiversity worldwide, strongly suggesting that we should reconcile urban development with conservation. Urbanization can follow two extreme types of development within a continuum: land sharing (buildings mixed with dispersed green space) or land sparing (buildings interspersed with green patches that concentrate biodiversity-supporting vegetation). Recent local-scale studies indicate that biodiversity is typically favored by land sparing. We investigated which of these two types of urbanization is associated with a higher taxonomic (i.e. species richness), functional, and phylogenetic diversity of birds. To do so, we collected information on breeding and wintering bird assemblages in 45 land-sharing and 45 land-sparing areas in nine European cities, which provide the first attempt to explore this question using a large geographical scale and temporal replication. We found that land-sharing urban areas were significantly associated with a higher taxonomic and functional diversity of birds during winter, but not during the breeding season (with only a marginally significant effect for functional diversity). We found no association between the type of urban development and phylogenetic diversity. Our findings indicate that not all components of avian diversity are similarly affected by these two means of urban planning and highlight the importance of integrating the temporal perspective into this kind of studies. Our results also offer useful information to the current debate about the trade-off between biodiversity conservation and human well-being in the context of land sharing and sparing urban practices. In addition, we found that certain small-scale urban landscape characteristics (i.e. few impervious surfaces, high water or tree cover) and human practices (i.e. bird feeders or plants with berries) can help maintaining more diverse urban bird assemblages. We provide specific suggestions for both policymakers and citizens that hopefully will help to create more biodiversity-friendly cities in the future.

Land-sharing vs. land-sparing urban development modulate predator-prey interactions in Europe.

Urban areas are expanding globally as a consequence of human population increases, with overall negative effects on biodiversity. To prevent the further loss of biodiversity, it is urgent to understand the mechanisms behind this loss to develop evidence-based sustainable solutions to preserve biodiversity in urban landscapes. The two extreme urban development types along a continuum, land-sparing (large, continuous green areas and high-density housing) and land-sharing (small, fragmented green areas and low-density housing) have been the recent focus of debates regarding the pattern of urban development. However, in this context, there is no information on the mechanisms behind the observed biodiversity changes. One of the main mechanisms proposed to explain urban biodiversity loss is the alteration of predator-prey interactions. Using ground-nesting birds as a model system and data from nine European cities, we experimentally tested the effects of these two extreme urban development types on artificial ground nest survival and whether nest survival correlates with the local abundance of ground-nesting birds and their nest predators. Nest survival (n = 554) was lower in land-sharing than in land-sparing urban areas. Nest survival decreased with increasing numbers of local predators (cats and corvids) and with nest visibility. Correspondingly, relative abundance of ground-nesting birds was greater in land-sparing than in land-sharing urban areas, though overall bird species richness was unaffected by the pattern of urban development. We provide the first evidence that predator-prey interactions differ between the two extreme urban development types. Changing interactions may explain the higher proportion of ground-nesting birds in land-sparing areas, and suggest a limitation of the land-sharing model. Nest predator control and the provision of more green-covered urban habitats may also improve conservation of sensitive birds in cities. Our findings provide information on how to further expand our cities without severe loss of urban-sensitive species and give support for land-sparing over land-sharing urban development.

Contagious fear: Escape behavior increases with flock size in European gregarious birds.

Flight initiation distance (FID), the distance at which individuals take flight when approached by a potential (human) predator, is a tool for understanding predator-prey interactions. Among the factors affecting FID, tests of effects of group size (i.e., number of potential prey) on FID have yielded contrasting results. Group size or flock size could either affect FID negatively (i.e., the dilution effect caused by the presence of many individuals) or positively (i.e., increased vigilance due to more eyes scanning for predators). These effects may be associated with gregarious species, because such species should be better adapted to exploiting information from other individuals in the group than nongregarious species. Sociality may explain why earlier findings on group size versus FID have yielded different conclusions. Here, we analyzed how flock size affected bird FID in eight European countries. A phylogenetic generalized least square regression model was used to investigate changes in escape behavior of bird species in relation to number of individuals in the flock, starting distance, diet, latitude, and type of habitat. Flock size of different bird species influenced how species responded to perceived threats. We found that gregarious birds reacted to a potential predator earlier (longer FID) when aggregated in large flocks. These results support a higher vigilance arising from many eyes scanning in birds, suggesting that sociality may be a key factor in the evolution of antipredator behavior both in urban and rural areas. Finally, future studies comparing FID must pay explicit attention to the number of individuals in flocks of gregarious species.

Territoriality and variation in home range size through the entire annual range of migratory great spotted cuckoos (Clamator glandarius).

Variation in home range size throughout the year and its causes are not well understood yet. Migratory brood parasites offer a unique opportunity to incorporate this spatio-temporal dimension into the study of the factors regulating home range dynamics. Using satellite transmitters, we tracked sixteen migratory great spotted cuckoos (Clamator glandarius) of both sexes for up to three years. We constructed home ranges in all major staging areas, from the Spanish breeding areas to the African wintering grounds, analyzed their temporal and geographical variation and investigated their main potential determinants (e.g. food and host availability). We found that home ranges were significantly larger in the breeding area compared to non-breeding areas. Using NDVI as a proxy for food availability, we showed that breeding area home ranges have significantly lower food availability per km2 than home ranges elsewhere which could explain why cuckoos use alternative areas with higher food availability before initiating migration. We also found some evidence for sex differences. Additionally, we found no indications of territoriality in this species, providing novel information into the current debate on brood parasite territoriality. Overall, food availability seems to be an important factor regulating home range dynamics and influencing migratory patterns throughout the year in great spotted cuckoos.

Migration behavior and performance of the great spotted cuckoo (Clamator glandarius).

The study of brood parasitism has traditionally been focused on the breeding period, but recent evidence suggests that it urgently needs a new spatio-temporal perspective to explore novel avenues on brood parasite-host co-evolutionary interactions. Many brood parasites are migrants, but their ecology outside their short breeding season is poorly known. The great spotted cuckoo (Clamator glandarius) is one of the classical models in the study of brood parasitism, however, there is very little information on its migratory strategy, route and wintering grounds. Furthermore, there is no previous information on the geographical distribution of mortality and its causes in this species; information that is critical to understand the fluctuations in cuckoo populations and detect potential conservation risks. Using satellite tracking technology, we provide novel insight into the migratory behavior and performance of the great spotted cuckoo. We found individuals from southern Spain to be long-distance nocturnal migrants that use the East Atlantic Flyway for both post and pre-breeding migration, and that winter in the western Sahel. We found evidence of individual variation in their migration route, particularly regarding their post-breeding behavior in Spain. Our study also suggests that the south of Morocco is the most dangerous area due to a large number of deaths during the post-breeding migratory period. Furthermore, we found that natural predation seems to be the main cause of death, probably due to raptors, although human activities (i.e. hunting) could also played a role in the southern Mediterranean shore. Our study offers novel findings and challenges traditional ideas on the ecology of this species providing a good example of how the new spatio-temporal perspective can expand our knowledge on brood parasites.

Nest predation risk modifies nestlings' immune function depending on the level of threat.

Predation risk is thought to modify the physiology of prey mainly through the stress response. However, little is known about its potential effects on the immunity of animals, particularly in young individuals, despite the importance of overcoming wounding and pathogen aggression following a predator attack. We investigated the effect of four progressive levels of nest predation risk on several components of the immune system in common blackbird (Turdus merula) nestlings by presenting them with four different calls during 1 h: non-predator calls, predator calls, parental alarm calls and conspecific distress calls to induce a null, moderate, high and extreme level of risk, respectively. Nest predation risk induced an increase in ovotransferrin, immunoglobulin and the number of lymphocytes and eosinophils. Thus, the perception of a potential predator per se could stimulate the mobilization of a nestling's immune function and enable the organism to rapidly respond to the immune stimuli imposed by a predator attack. Interestingly, only high and extreme levels of risk caused immunological changes, suggesting that different immunological parameters are modulated according to the perceived level of threat. We also found a mediator role of parasites (i.e. Leucocytozoon) and the current health status of the individual, as only nestlings not parasitized or in good body condition were able to modify their immune system. This study highlights a previously unknown link between predation risk and immunity, emphasizing the complex relationship among different selective pressures (predation, parasitism) in developing organisms and accentuating the importance of studying predation from a physiological point of view.

Urban blackbirds have shorter telomeres.

Urbanization, one of the most extreme human-induced environmental changes, represents a major challenge for many organisms. Anthropogenic habitats can have opposing effects on different fitness components, for example, by decreasing starvation risk but also health status. Assessment of the net fitness effect of anthropogenic habitats is therefore difficult. Telomere length is associated with phenotypic quality and mortality rate in many species, and the rate of telomere shortening is considered an integrative measure of the 'life stress' experienced by an individual. This makes telomere length a promising candidate for examining the effects of urbanization on the health status of individuals. We investigated whether telomere length differed between urban and forest-dwelling common blackbirds (Turdus merula). Using the terminal restriction fragment assay, we analysed telomere length in yearlings and older adults from five population dyads (urban versus forest) across Europe. In both age classes, urban blackbirds had significantly shorter telomeres (547 bp) than blackbirds in natural habitats, indicating lower health status in urban blackbirds. We propose several potential hypotheses to explain our results. Our findings show that even successful city dwellers such as blackbirds pay a price for living in these anthropogenic habitats.

Bacterial density rather than diversity correlates with hatching success across different avian species.

Bacterial communities within avian nests are considered an important determinant of egg viability, potentially selecting for traits that confer embryos with protection against trans-shell infection. A high bacterial density on the eggshell increases hatching failure, whether this effect could be due to changes in bacterial community or just a general increase in bacterial density. We explored this idea using intra- and interspecific comparisons of the relationship between hatching success and eggshell bacteria characterized by culture and molecular techniques (fingerprinting and high-throughput sequencing). We collected information for 152 nests belonging to 17 bird species. Hatching failures occurred more frequently in nests with higher density of aerobic mesophilic bacteria on their eggshells. Bacterial community was also related to hatching success, but only when minority bacterial operational taxonomic units were considered. These findings support the hypothesis that bacterial density is a selective agent of embryo viability, and hence a proxy of hatching failure only within species. Although different avian species hold different bacterial densities or assemblages on their eggs, the association between bacteria and hatching success was similar for different species. This result suggests that interspecific differences in antibacterial defenses are responsible for keeping the hatching success at similar levels in different species.

Fecal sacs do not increase nest predation in a ground nester.

Most altricial birds remove their nestlings' feces from the nest, but the evolutionary forces driving this behavior are poorly understood. A possible adaptive explanation for this could be that birds avoid the attraction of nest predators to their nests due to the visual or olfactory cues produced by feces (nest predation hypothesis). This hypothesis has received contrasting support indicating that additional experimental studies are needed, particularly with respect to the visual component of fecal sacs. To test this hypothesis, we conducted an experiment manipulating the presence of fecal sacs on inactive Woodlark (Lullula arborea) nests. This ground nester has highly cryptic nests that are mainly depredated by visually oriented nest predators (i.e., corvids) in our study population, making it an excellent system to test for the nest predation hypothesis. Our results showed that the presence of fecal sacs in the nest does not seem to be an important factor explaining nest predation. Interestingly, the effect of nest concealment, the most important factor explaining nest predation in Woodlark nests, depended on whether the nest was depredated the previous year or not, supporting the importance of using different nesting sites between years. Our findings indicate that this important nest sanitation behavior is not likely motivated by nest predation and highlight the need to explore alternative selective pressures in this context.