Revealing the roles of egg darkness and nest similarity for a cryptic parasite egg versus host's cognition: an alternate coevolutionary trajectory.

As a text-book example of coevolution, the escalating interactions between egg mimicry by parasitic cuckoos and egg recognition by their hosts constitute a key battlefield for parasitism and anti-parasitism strategies. However, some parasite-host systems have deviated from this coevolutionary trajectory because some cuckoos do not lay mimetic eggs, while the hosts do not recognize them, even under the high costs of parasitism. The cryptic egg hypothesis was proposed to explain this puzzle, but the evidence to date is mixed and the relationship between the two components of egg crypticity, egg darkness (dim egg coloration) and nest similarity (similarity to host nest appearance), remains unknown. Here, we developed a 'field psychophysics' experimental design to dissect these components while controlling for undesired confounding factors. Our results clearly show that both egg darkness and nest similarity of cryptic eggs affect recognition by hosts, and egg darkness plays a more influential role than nest similarity. This study provides unambiguous evidence to resolve the puzzle of absent mimicry and recognition in cuckoo-host systems and explains why some cuckoo eggs were more likely to evolve dim coloration rather than similarity to host eggs or host nests.

Defensive adaptations to cuckoo parasitism in the black-browed reed warbler (Acrocephalus bistrigiceps): recognition and mechanism.

Hosts of avian brood parasites suffer a high cost of reproductive loss due to parasitism, driving them to evolve a variety of anti-parasitic defenses. These defenses comprise a series of components, including the recognition of brood parasites and the eggs laid by the parasites, cues used for recognition, and the mechanisms on which these behaviors are based. In this study, we conducted egg recognition and nest intruder experiments to examine these components of anti-parasitic behavior in the black-browed reed warbler (Acrocephalus bistrigiceps), a rare host of the common cuckoo (Cuculus canorus). We found that the host possessed strong recognition capacity, rejecting 100% of parasitic eggs, and used a template-based mechanism for egg recognition. The host birds also rejected 80% of their own eggs on which artificial markings were added to the blunt pole; however, they accepted all eggs with the same manipulation on the sharp pole, implying that the blunt pole was an important recognition cue. Furthermore, the host exhibited stronger aggression to cuckoos than to harmless controls; a behavior specific to the incubation stage rather than the nestling stage. Therefore, the host was able to distinguish the cuckoo from other nest intruders as being a brood parasite. These results together help explain the near absence of cuckoo parasitism in black-browed reed warblers and provide new information concerning anti-parasitic defenses in this host species.

Nest size matters: common cuckoos prefer to parasitize larger nests of Oriental reed warblers.

Avian brood parasites leave parental care of their offspring to foster parents. Theory predicts that parasites should select for large host nests when they have sufficient available host nests at a given time. We developed an empirical experimental design to test cognitive ability of female cuckoos in nest size by studying nest choice of common cuckoos (Cuculus canorus) among nests of its Oriental reed warbler (Acrocephalus orientalis) hosts. We presented three groups of experimental nests: 1) nest dyads tied together including one large and one small artificial nest from reed leaves, 2) nest triads tied together used the old modified warbler's own nests including enlarged, reduced and medium-sized nests, and 3) nest dyads are similar to group 1, but not tied together to elicit parasitism by common cuckoos. We predict that cuckoos prefer larger nest than medium one, the next is smaller nest. Our findings showed that common cuckoo females generally prefer large nests over medium or small sized nests in all three experimental groups. Furthermore, cuckoo parasitism was significantly more common than in previous studies of the same warbler population, implying that larger, higher and more exposed host nests effectively increased the probability of cuckoo parasitism.

Cuckoos use host egg number to choose host nests for parasitism.

To maximize their offspring success common cuckoo (Cuculus canorus) females should lay their eggs into host nests before incubation has begun. This ensures that the parasite chick hatches before all host chicks and can evict its foster siblings to monopolize host parental care. Many studies have demonstrated that most cuckoo eggs are indeed laid before the onset of host incubation. But cues used by female cuckoos to choose the right nest at the right time remain unclear. Here, we combine field observations with a field experiment to test whether female cuckoos use the number of eggs in the nests of their Oriental reed warbler (Acrocephalus orientalis) hosts to direct their choice. Over 8 years of field observations and 5 years of experiments, cuckoo females placed the majority of their eggs in nests with fewer than three host eggs, i.e. early in the laying sequence. For natural nests, the cuckoos may use information gleaned from the activity and behaviour of the host parents to make their choice. In our sets of experimental nests containing different numbers of model eggs, the vast majority of parasitism events occurred in nests containing a single egg. To our knowledge, this is the first field experiment showing that cuckoos choose host nests for parasitism based on the number of host eggs they contain. It appears that cuckoo females use the egg number to estimate the appropriate host nest stage for timely parasitism.

Model eggs fail to detect egg recognition in host populations after brood parasitism is relaxed.

BACKGROUND: Obligate brood parasites exert strong selective pressure on target hosts. In response, hosts typically evolve anti-parasitism strategies, of which egg recognition is one of the most efficient. Generally, host egg-recognition capacity is determined using model eggs. Previous studies have shown that some host species, which are capable of detecting parasite eggs, do not reject model eggs. However, it is unknown that whether the reaction to model eggs varies among distinct populations of the same host in relation to the degree of parasitism pressure. RESULTS: Here, we compared the rejection frequencies of model eggs and real eggs between mainland and island populations of the plain prinia (Prinia inornata), which are respectively sympatric and allopatric with their brood parasite, the common cuckoo (Cuculus canorus). Our results indicated that the mainland and island populations rejected real eggs at similar rates, but rejected model eggs, which were similar in size to real eggs but heavier, at significantly different rates: the island population rejected fewer model eggs, possibly because the rejection motivation of this population was lower due to absence of parasitism. CONCLUSIONS: Our results indicated that some factors affecting the decision to reject, such as rejection motivation, varied according to the degree of parasitism pressure, and thus influenced the frequency of egg rejection. Furthermore, our results suggested that model eggs should be used with caution in comparative studies of egg recognition abilities among species or populations subjected to different intensities of brood parasitism. That is, model eggs may fail to accurately detect egg recognition in host populations with little to no risk of parasitism.

Keeping eggs warm: thermal and developmental advantages for parasitic cuckoos of laying unusually thick-shelled eggs.

Obligate brood parasites have evolved unusually thick-shelled eggs, which are hypothesized to possess a variety of functions such as resistance to puncture ejection by their hosts. In this study, we tested the hypothesis that obligate brood parasites lay unusually thick-shelled eggs to retain more heat for the developing embryo and thus contribute to early hatching of parasite eggs. By doing so, we used an infrared thermal imaging system as a non-invasive method to quantify the temperature of eggshells of common cuckoos (Cuculus canorus) and their Oriental reed warbler (Acrocephalus orientalis) hosts in an experiment that artificially altered the duration of incubation. Our results showed that cuckoo eggshells had higher temperature than host eggs during incubation, but also less fluctuations in temperature during incubation disturbance. Therefore, there was a thermal and hence a developmental advantage for brood parasitic cuckoos of laying thick-shelled eggs, providing another possible explanation for the unusually thick-shelled eggs of obligate brood parasites and earlier hatching of cuckoo eggs compared to those of the host.

Nest sanitation behavior in hirundines as a pre-adaptation to egg rejection to counter brood parasitism.

Previous studies suggested that nest sanitation behavior may have been a pre-adaptation from which egg rejection of brood parasite eggs evolved. We tested this hypothesis in two swallow species, the red-rumped swallow (Cecropis daurica) and the barn swallow (Hirundo rustica). Our results indicated that the red-rumped swallow, which is an accepter of foreign eggs, rejected a low percentage of non-egg-shaped objects and did so less often than the barn swallow, which is an intermediate rejecter of foreign eggs. Furthermore, the egg rejection rates of the barn swallow increased with the increase in rejection rates of non-egg-shaped objects among different populations. These results showed that nest cleaning behavior could have evolved into a means of reducing the costs of brood parasitism, suggesting that egg recognition ability has evolved from recognition of non-egg-shaped objects. This finding advances our understanding of the evolution of egg recognition behavior in birds.

Nest sanitation elicits egg discrimination in cuckoo hosts.

Nest sanitation is a nearly universal behavior in birds, while egg discrimination is a more specific adaptation that has evolved to counter brood parasitism. These two behaviors are closely related with nest sanitation being the ancestral behavior, and it has been hypothesized to constitute a preadaptation for egg discrimination. However, previous studies found little evidence to support this hypothesis. Here, we conducted an empirical test of the association between nest sanitation and egg discrimination in the barn swallow (Hirundo rustica) by inserting a single non-mimetic model egg or a non-mimetic model egg plus half a peanut shell into host nests. Compared to the rejection rate of single model eggs, barn swallows significantly increased egg rejection frequency if a half peanut shell was simultaneously introduced. Our result for the first time shows the impact of nest sanitation on egg discrimination and demonstrates that nest sanitation can elicit egg discrimination in hosts of brood parasites. This study provided evidence for nest sanitation being a preadaptation to egg discrimination by facilitating egg rejection, thereby significantly advancing our understanding of avian cognition of foreign objects. Furthermore, we suggest that egg discrimination behavior in many accepters and intermediate rejecters may be lost or diluted. Such egg discrimination can be elicited and restored after nest sanitation, implying a sensitive and rapid phenotypic response to increased risk of parasitism. Our study offers a novel perspective for investigating the role of so-called intermediate rejecter individuals or species in the long-term coevolutionary cycle between brood parasites and their hosts.

Climate niche differentiation between two passerines despite ongoing gene flow.

Niche evolution underpins the generation and maintenance of biological diversity, but niche conservatism, in which niches remain little changed over time in closely related taxa, and the role of ecology in niche evolution are continually debated. To test whether climate niches are conserved in two closely related passerines in East Asia - the vinous-throated (Paradoxornis webbianus) and ashy-throated (P. alphonsianus) parrotbills - we established their potential allopatric and sympatric regions using ecological niche models and compared differences in their climate niches using niche overlap indices in background tests and multivariate statistical analyses. We also used polymorphism data on 44 nuclear genes to infer their divergence demography. We found that these two parrotbills occupy different climate niches, in both their allopatric and potential sympatric regions. Because the potential sympatric region is the area predicted to be suitable for both parrotbills based on the ecological niche models, it can serve as a natural common garden. Therefore, their observed niche differences in this potential sympatry were not simply rendered by phenotypic plasticity and probably had a genetic basis. Our genetic analyses revealed that the two parrotbills are not evolutionarily independent for the most recent part of their divergence history. The two parrotbills diverged c. 856,000 years ago and have had substantial gene flow since a presumed secondary contact c. 290,000 years ago. This study provides an empirical case demonstrating that climate niches may not be homogenized in nascent species in spite of substantial, ongoing gene flow, which in turn suggests a role for ecology in promoting and maintaining diversification among incipient species.

Nest defenses and egg recognition of yellow-bellied prinia against cuckoo parasitism.

Parasites may, in multi-parasite systems, block the defenses of their hosts and thus thwart host recognition of parasites by frequency-dependent selection. Nest defenses as frontline may block or promote the subsequent stage of defenses such as egg recognition. We conducted comparative studies of the defensive strategies of a host of the Oriental cuckoo Cuculus optatus, the yellow-bellied prinia Prinia flaviventris, in mainland China with multiple species of cuckoos and in Taiwan with a single cuckoo species. Cuckoo hosts did not exhibit aggression toward cuckoos in the presence of multiple cuckoo species but showed strong aggressive defenses of hosts directed toward cuckoos in Taiwan. Furthermore, the cuckoo host in populations with a single cuckoo species was able to distinguish adults of its brood parasite, the Oriental cuckoo, from adult common cuckoos (Cuculus canorus). This represents the first case in which a cuckoo host has been shown to specifically distinguish Oriental cuckoo, from other Cuculus species. Hosts ejected eggs at a higher rate in a single cuckoo species system than in a multi-species cuckoo system, which supports the strategy facilitation hypothesis. Granularity analysis of variation in egg phenotype based on avian vision modeling supported the egg signature hypothesis in hosts because Taiwanese prinias increased consistency in the appearance of their eggs within individual hosts thus favoring efficient discrimination against cuckoo eggs. This study significantly improves our knowledge of intraspecific variation in antiparasitism behavior of hosts between single- and multi-cuckoo systems.

Dynamics of evolutionary succession and coordination between opposite adaptations in cuckoo hosts under antagonistic coevolution.

Adaptations are driven by specific natural selection pressures throughout biological evolution. However, these cannot inherently align with future shifts in selection dynamics, thus manifesting in opposing directions. We performed field experiments on cuckoo hosts to investigate the coexistence and conflict between two evolutionarily successive but opposing behavioral adaptations-egg retrieval and rejection. Our findings provide key insights. (1) Egg rejection against brood parasites in hosts reshapes egg retrieval to flexible reactions-retrieval, ignoring, or outright rejection of foreign eggs outside the nest cup, departing from instinctual retrieval. (2) Parasitism pressure and egg mimicry by parasites remarkably alter the proportions of the three host reactions. Host species with higher parasitism pressure exhibit frequent and rapid rejection of non-mimetic foreign eggs and reduced ignoring or retrieval responses. Conversely, heightened egg mimicry enhances retrieval behaviors while diminishing ignoring responses. (3) Cuckoos employ consistent mechanisms for rejecting foreign eggs inside or outside the nest cup. Direct rejection of eggs outside the nest cup shows that rejection precedes retrieval, indicating prioritization of specific adaptation over instinct. (4) Cuckoo hosts navigate the conflict between the intentions and motivations associated with egg rejection and retrieval by ignoring foreign eggs, a specific outcome of the rejection-retrieval tradeoff.

Comparison of Reproductive Strategies between Two Sympatric Copsychus Passerines.

Reproduction plays a crucial role in determining the development, fate, and dynamics of bird populations. However, reproductive strategies vary among species and populations. In this study, we investigated the reproductive strategies of the Oriental Magpie Robin (Copsychus saularis) and White-rumped Shama (C. malabarica), which are closely related passerines that reproduce in sympatric areas. We found that although these two species were both cavity nesting, their nest-site selection differed; the Shama preferred nesting close to trees and forests, whereas the Magpie Robin nested close to human residential areas. Furthermore, their egg incubation patterns differed; the Shama increased daily incubation frequency with incubation time, but the Magpie Robin maintained its daily incubation time regardless of changes in incubation frequency. However, the nestling heating patterns of these two species were similar, indicating a critical demand for regulating hatchling body temperature during this crucial stage. The feeding frequencies of male parents were strongly correlated with those of females in both species, suggesting equal contribution and good synchronization between the sexes. Nestling feeding frequency was also correlated with nest cleaning frequency, implying coordination between feeding and defecation by parents and offspring, respectively. This research explored the divergence and convergence of reproductive strategies between these two sympatric species, providing valuable insights into the niche differentiation theory.

Illegal and Exploitative Sand-Digging Activities Could Be Managed to Create Suitable Nesting Habitats for Blue-Tailed Bee-Eaters (Merops philippinus).

The development of human society often interferes with wild animals and their natural habitats. Interference during resource exploitation is mostly negative; however, in some cases, it can be positive and even have significance for some species. In this study, we investigated the number of blue-tailed bee-eaters (Merops philippinus), a species under 'state protection category II' in China, between controlled and manipulated nesting habitats from 2017 to 2022. Our results indicated that commercial sand-digging activities, either illegal or approved, initially created suitable nesting habitats to attract blue-tailed bee-eaters but subsequently led to damage of nests or nesting habitats. However, sand digging can be modified by avoiding the breeding season to provide safe and suitable nesting habitats for bee-eaters. The number of breeding birds more than tripled when digging during the breeding season was avoided. We also found that conventional conservation strategies, which strictly prohibited sand-digging activities, did not contribute to the nesting habitats of bee-eaters. This study enriches the theories of conservation biology and emphasizes the importance of dialectical thinking regarding exploitative and seemingly destructive activities.

Egg rejection based on egg size recognition as a specific strategy against parasitic cuckoos.

In the coevolutionary interactions between avian brood parasites and their hosts, egg recognition based on color and/or pattern is a common and effective defense to counter parasitism. However, for egg recognition based on size, only a few studies have found affirmative results, and they do not provide unambiguous evidence that egg size recognition in hosts has evolved as an important and specific anti-parasite adaptation against parasite eggs. We studied the brood parasite system between the Asian emerald cuckoo Chrysococcyx maculatus and its host, the chestnut-crowned warbler Phylloscopus castaniceps. The cuckoo parasitizes the warbler using non-mimetic and larger eggs at a parasitism rate of 12.9%. The warbler nests used in this experiment were built in a dark environment with the nest illuminance near 0 lux. Experiments with 2 types of model eggs with colors and patterns resembling cuckoo eggs of different sizes (cuckoo egg size or host egg size) showed that the warblers were able to reject 63.6% of cuckoo model eggs under these dim light conditions. However, model eggs with the same color and pattern similar to the warbler egg size were always accepted. This study provides strong evidence supporting the theory that egg size recognition can be evolved in hosts as a specific anti-parasite adaptation against cuckoos. We suggest that the egg size recognition of the warbler is an outcome of the tradeoff between the costs of violating the parental investment rule and suffering cuckoo parasitism.

Variation in russet sparrow (Passer cinnamomeus) breeding biology in relation to small-scale altitudinal differences in China.

Evolution of avian life histories is typically strongly influenced by both altitude and latitude. To date, most studies have investigated the effects of extreme differences in altitude and latitude on variation in reproductive traits. Studies based on small altitude and latitude spans are needed to better understand the resolution of selective pressures. We compared several aspects of russet sparrow (Passer cinnamomeus) breeding biology between a low-altitude (200 m) and a high-altitude (1,500 m) population in China, representing a relatively small altitudinal gradient (1,300 m). High-altitude birds initiated breeding significantly later compared to their low-altitude counterparts. Interestingly, breeding season was significantly longer in the high-altitude site (57 vs. 84 d). Lowland sparrows laid larger clutches (4.92 vs. 4.09 eggs) and showed greater fledging success (4.20 vs. 3.46 fledgings) than did upland birds. Variation in life history traits thus appears to occur even along a small scale altitudinal gradient. We suggest that the longer breeding season and smaller clutch size in the highland population may be an adaptation or acclimation to compensate for the reduced annual productivity resulting from unfavourable or sub-optimal habitats for these sparrows.

Anti-Predation Responses to Conspecific versus Heterospecific Alarm Calls by the Nestlings of Two Sympatric Birds.

Predation is generally the main cause of bird mortality. Birds can use acoustic signals to increase their predation survival. Bird response to mobbing alarm calls is a form of anti-predation behavior. We used a playback technique and acoustic analysis to study the function of mobbing alarm calls in the parent-offspring communication of two sympatric birds, the vinous throated parrotbill (Sinosuthora webbianus) and oriental reed warbler (Acrocephalus orientalis). The chicks of these two species responded to conspecific and heterospecific mobbing alarm calls by suppressing their begging behavior. The mobbing alarm calls in these two species were similar. Mobbing alarm calls play an important role in parent-offspring communication, and chicks can eavesdrop on heterospecific alarm calls to increase their own survival. Eavesdropping behavior and the similarity of alarm call acoustics suggest that the evolution of alarm calls is conservative and favors sympatric birds that have coevolved to use the same calls to reduce predation risk.

Female Cuckoo Calls Deceive Their Hosts by Evoking Nest-Leaving Behavior: Variation under Different Levels of Parasitism.

The common cuckoo (Cuculus canorus) is an obligate brood parasite that has evolved a series of strategies to trick its hosts. The female cuckoo has been hypothesized to mimic the appearance and sounds of several raptors to deceive the hosts into exhibiting anti-predator behavior. Such behavior would relax the protection of the host nest and thus allow the female cuckoo to approach the host nest unopposed. Many anti-parasite strategies have been found to vary among geographical populations due to different parasitic pressures from cuckoos. However, the effect of female cuckoo calls related to different levels of parasitic pressure has not been examined. Here, we studied the effect of female cuckoo calls on the oriental reed warbler (Acrocephalus orientalis), one of the major hosts of the common cuckoo, in two geographical populations experiencing different levels of parasite pressure. Four kinds of sounds were played back to the hosts: the calls from female common cuckoos, male common cuckoos, sparrowhawks (Accipiter nisus), and oriental turtle doves (Streptopelia orientalis). The results showed that the female cuckoo calls induced the hosts to leave their nests more frequently than the male cuckoo or dove calls in both populations, and two populations of the hosts reacted similarly to the female cuckoo calls, implying that the function of female cuckoo calls would not be affected by the difference in parasitism rate. This study indicates that female cuckoo calls function to distract the hosts' attention from protecting their nests. However, we propose that such a deception by the female cuckoo call may not be due to the mimicry of sparrowhawk calls, but rather that the rapid cadence of the call that causes a sense of anxiety in the hosts.

Parent-offspring and inter-offspring responses to conspecific versus heterospecific distress calls in 2 sympatric birds.

Distress calls, as a type of alarm call, play important roles in expressing bodily condition and conveying information concerning predation threats. In this study, we examined the communication via distress calls in parent-offspring and inter-offspring interactions. First, we used playback of chick distress calls of 2 sympatric breeders, the vinous-throated parrotbill Sinosuthora webbiana and the oriental reed warbler Acrocephalus orientalis, to the adults/chicks of these 2 species, respectively, and measured the responses of conspecifics or heterospecifics. The playback-to-chicks experiment showed that both species of chicks reduced the number of begging calls and begging duration time as a response to conspecific/heterospecific distress calls compared with natural begging and background noise controls. However, reed warbler chicks also reduced beak opening frequency in the response to conspecific distress calls compared with other playback stimuli. Second, the results of the playback-to-adults experiment showed that reed warbler adults could eavesdrop on distress calls of conspecific neighbors and sympatric heterospecifics. Furthermore, the nest-leaving behavior of reed warblers did not differ significantly when they heard the distress calls of conspecifics or parrotbills. Finally, reed warbler adults responded to heterospecific distress calls more quickly than to conspecific distress calls, and parrotbill adults presented the same response. Our results supported the warn-kin hypothesis and show that chick distress calls play an important role in conveying risk and the condition of chicks to enhance individual fitness. In addition, we also found that eavesdropping on distress calls is a congenital behavior that begins in the chick stage.

Egg Rejection and Nest Sanitation in an Island Population of Barn Swallows (Hirundo rustica): Probability, Response Latency, and Sex Effects.

Bird nests function as vessels for eggs and nestlings, and an environment for rearing offspring. However, foreign objects falling into bird nests and nestling eggshells may be harmful. Moreover, the smell of fecal sacs increases the risk of detection by predators. Many bird species have evolved nest sanitation to prevent damage to their nests. Furthermore, egg rejection evolved in some birds to thwart brood parasites that lay eggs in their nests. We studied 133 nests of barn swallows (Hirundo rustica) in an island population through a nest content manipulation experiment to determine nest sanitation and egg rejection behaviors and their relationship. Swallows rejected non-egg foreign objects more frequently (100% vs. 58.6%) and sooner than parasite eggs, which supports the hypothesis that nest sanitation is a pre-adaptation to egg rejection. However, nest sanitation did not increase egg rejection, either in probability or latency. Furthermore, both sexes incubated the eggs, cleaned the nests, and removed parasite eggs, implying that both are confronted with natural selection related to nest sanitation and brood parasitism. However, females invested more time in these behaviors than males. This provides evidence for the evolutionary relationship of nest sanitation and egg rejection behaviors in barn swallows.