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.

Cryptic eggs are rejected less frequently by a cuckoo host.

Obligate brood parasitism is associated with huge reproduction costs, forcing hosts to evolve various anti-parasitic strategies against brood parasites, among which egg recognition and rejection is the most effective defense strategy. According to the crypsis hypothesis, non-mimetic yet cryptic eggs in a nest can also deceive their hosts and eventually be accepted. To validate this hypothesis, we conducted field experiments on Oriental reed warblers (Acrocephalus orientalis), a common host for common cuckoos (Cuculus canorus). We firstly tested the egg recognition and rejection abilities of Oriental reed warblers, using black and white model eggs in natural nests. Then we designed a comparison test where the cryptic effects of the two groups of experimental eggs were different. We manipulated the nest lining color and added relatively cryptic and bright model eggs to test warblers' rejection behaviors against cryptic and bright foreign eggs. The results showed that warblers have strong egg recognition and rejection abilities. There is a significant tendency for warblers to prefer to peck and reject relatively distinguishable foreign eggs, which supports the crypsis hypothesis. These findings indicate that even in the host-parasite system of open nests, parasitic eggs that are cryptic enough are prevented from being discovered and rejected by the host, and thus obtain the possibility of successful 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.

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.

Three sympatric host nestlings eavesdrop on cuckoo nestling distress calls.

In predator-prey interactions, the prey faces extreme challenges from predation, which drives the evolution of defense or anti-predator mechanisms. Compared with adult birds, nestlings are more vulnerable but not helpless. However, data on whether nestlings eavesdrop on the danger signals transmitted by other prey nestlings and the mechanisms of eavesdropping remain limited. In brood parasitism, common cuckoo (Cuculus canorus) nestlings, raised by host adults who are not closely related, offer an instructive system for studying the transmission and recognition of danger signals among nestlings of different species that share special relationships. We played back the distress calls of common cuckoo nestlings to nestlings of three sympatric host species (the oriental reed warbler Acrocephalus orientalis, which is a primary host of the common cuckoo, the reed parrotbill Paradoxornis heudei, an occasional host, and the vinous-throated parrotbill Sinosuthora webbiana, which is not parasitized in the study area) to investigate whether the host nestlings reduced their begging behavior. We also quantified the degree of inhibition toward begging behavior for these nestlings. The results revealed that, in response to the distress calls, the three sympatric host species markedly suppressed their begging behavior. This response can likely be attributed to the innate response of host nestlings caused by the general characteristics of distress calls, rather than the acoustic similarity and phylogenetic relationship between host nestlings and cuckoo nestlings. Furthermore, we observed that upon hearing the distress calls of cuckoo nestlings, the oriental reed warbler nestlings exhibited the greatest reduction in the total number of calls compared to the other two host species, potentially owing to stronger predation and parasitic pressures. This study suggests that host nestlings can detect danger signals emitted by parasitic nestlings; however, further investigation is needed to determine whether they can respond to distress calls from unfamiliar nestlings in different regions.

[Research on optimization of imaging system of the hand vein optical properties].

Due to the difficulties of the copying, vein identification has developed rapidly in recent years. The light source selection directly affects the image quality. This paper acquired by experiment the reflectivities of vein and non-vein irradiation with different wavelengths of near infra-red. Comparing the strength of reflectivities of various wave lengths, we found that there were the strongest contrasts between vein and non-vein in the 810 nm, and 810 nm near infra-red was suitable to a vein imaging light source. Finally, clear hand vein images were obtained with the selected light source.

Importance of cooperation: How host nest defenses effectively prevent brood parasitism from the cuckoos.

As the frontline defense against avian brood parasitism, nest defense is important in reducing nest parasitism and increasing host fitness. However, systematic studies on its effectiveness (i.e., on whether it successfully prevents cuckoo parasitism) are scarce. Based on 214 video recordings from 10 years of field observations, we evaluated the effectiveness of nest defenses of the Oriental reed warbler (Acrocephalus orientalis) in deterring common cuckoo (Cuculus canorus) parasitism. Under a breeding pair situation (≤ 2 hosts individuals), warblers were hardly effective in resisting the egg-laying of the cuckoo and many of the cuckoos under attack were able to complete the parasitic process without being harmed. However, when hosts ≥ 3 individuals, the warblers could effectively thwart the cuckoos and cause lethal damage to the cuckoo in partial cases. This indicated that the host group effectively resisted the cuckoo, demonstrating the importance of cooperation among host neighbors in resisting cuckoo parasitism.

Predation of Daurian redstarts offspring in nest boxes by the Oriental magpie-robin and tree sparrow.

Birds select suitable nest sites for breeding to ensure their own and offspring's survival; however, they inevitably suffer some potential predation risk. We studied the breeding ecology of Daurian redstarts (Phoenicurus auroreus) by providing nest boxes for their breeding from March to August of 2022. We recorded the predation of both Daurian redstarts eggs or nestlings by Oriental magpie-robins (Copsychus saularis) and tree sparrow (Passer montanus). Oriental magpie-robin were recorded attacking a feeding female adult and depredating nestlings. After the nestling predation event, the Daurian redstarts abandoned the nest. This video evidence provide a better understanding of the potential predators of cavity-nesting birds.

First description of injury feigning behavior in Godlewski's buntings.

Nest defense behavior helps ground-nesting birds improve their breeding success. Among such behaviors, injury feigning behavior (IFB) is one of the better ways to attract predators and protect birds' nests. IFB is generally associated with shorebirds in general and plovers in particular, however, through field observation, it has been found this behavior is also exhibited in bunting species when they facing the risk of predation. We attempt to explore the detailed description of this behavior in buntings and the factors that affect this behavior. Based on video-recordings of the nest defense behavior by the ground-nesting bird Godlewski's bunting (Emberiza godlewskii) against nest invaders, we explored the characteristic of IFB and differences in nest defense behavior during their brooding seasons. The results showed that female buntings had a distinct IFB: the wings of buntings incited within 60°-90° of the body, ran and called rapidly, pretending to be injured and unable to fly. The nest stage had significant and extremely significant effects on IFB and movement distance (MD), respectively. And IFB was more likely to occur in brooding (34.62%) than incubation period (16.42%). This indicates that there are significant differences in the IFB of buntings at different breeding stages.

Defense behavior of two closely related but geographically distant host species against cuckoo parasitism: A next test for the parallel coevolution.

Interactions between avian brood parasites, such as common cuckoos (Cuculus canorus), and their hosts are one of the best-studied examples of the coevolutionary arms race. Different stages of this arms race can be seen in different races of common cuckoos and their hosts across their range. However, little is known whether selected populations of two closely related but geographically distant species with probably different coevolutionary histories with the common cuckoo are also at different stages of the arms race. In this study, we tested this prediction experimentally using the same non-mimetic model eggs and three-dimensional (3D) printed models of the gray adult common cuckoo (Cuculus canorus). We examined egg recognition and egg rejection and aggression against the common cuckoo in the great reed warbler (Acrocephalus arundinaceus) and Oriental reed warbler (Acrocephalus orientalis), in Slovakia of Europe and northeast China of Asia. The results showed that the great reed warbler exhibited stronger responses to experimental model eggs and 3D models of the common cuckoo than the Oriental reed warbler. We conclude that both the great reed warbler and Oriental reed warbler have well-developed antiparasitic behaviors against common cuckoos in the studied populations, but with different levels of defense intensity, which may be due to local differences in parasitic pressure and the risk of parasitism. This provides an opportunity to study coevolutionary processes between the brood parasite and its hosts together in both species at large geographical scales.

Why cuckoos remove host eggs: Biting eggs facilitates faster parasitic egg-laying.

Brood parasitism by cuckoos relies on manipulating hosts to raise their offspring and has evolved stunning adaptations to aid in their deception. The fact that cuckoos usually but not always, remove one or two host eggs while laying their eggs has been a longstanding focus of intensive research. However, the benefit of this behavior remains elusive. Moreover, the recently proposed help delivery hypothesis, predicting that egg removal by cuckoos may decrease the egg-laying duration in the parasitism process caused by biting action, lacks experimental verification. Therefore, in this study, we examined the effects of egg removal/biting on the egg-laying speed in the common cuckoo (Cuculus canorus) to experimentally test this hypothesis. We compared the duration of cuckoo egg-laying in empty nests, nests with host eggs, and nests with artificial blue stick models to test whether cuckoos biting an egg/stick can significantly hasten the egg-laying speed than no biting action. Our results showed that biting an egg or an object is associated with cuckoos laying approximately 37% faster than when they do not bite an egg or an object. This study provides the first experimental evidence for the help delivery hypothesis and demonstrates that when cuckoos bite eggs or other objects in the nest, they lay eggs more quickly and thereby avoid suffering the hosts' injurious attack.

Nest desertion as an anti-parasitism strategy in hosts selects for late egg-laying behavior in cuckoos.

Studies have shown that brood parasites lay their eggs early in the egg-laying sequence of their hosts, providing them with the advantage of earlier hatching. However, common cuckoos (Cuculus canorus) appear to parasitize the nests of gray bushchat (Saxicola ferreus) during the late egg-laying stage. The bushchat often abandons parasitized nests in the early stages, but not in the late egg-laying stages, thus favoring late egg-laying by cuckoos. In this study, four experiments were conducted to determine whether gray bushchats employ a nest desertion strategy targeted at cuckoo parasitism. The results showed that nest desertion was significantly correlated with parasitism cues and occurred mainly during the hosts' early egg-laying stage. Our study provides the first experimental evidence that nest desertion is an anti-parasitic strategy used by hosts in response to cuckoos. Additionally, our experiments demonstrated that the nest desertion is influenced by the trade-offs of investments in different egg-laying stages.

Brood parasitism and egg recognition in three bunting hosts of the cuckoos.

Comparative studies of egg recognition and rejection between various sympatric hosts provide insight into the coevolutionary history of the hosts and parasites, as well as the degree of antagonism between the species. Although buntings are widely considered to be a suitable host taxon for cuckoos, there has been relatively little research on this example of parasitism and host antiparasitic behaviour. Here we provided the first report on brood parasitism and egg recognition in three sympatric ground-nesting bunting hosts of the common cuckoo (Cuculus canorus), namely the yellow-throated bunting (Emberiza elegans), south rock bunting (E. yunnanensis), and crested bunting (E. lathami). The results show that for the five breeding seasons during 2018-2022, the parasitism rate by common cuckoos was 0.87% and 0.45% in yellow-throated buntings and south rock buntings, respectively, whereas the parasitism rate by an unidentified parasite was 4% during 2018-2023 in the crested bunting. The rejection rates of the three bunting hosts for blue non-mimetic eggs were 89.3%, 88.9%, and 100% for yellow-throated buntings, south rock buntings, and crested buntings, respectively. The rejection rates for red non-mimetic eggs by yellow-throated buntings and south rock buntings were lower at 76.9% and 82.4%, respectively. All three sympatric bunting hosts examined had high levels of egg recognition and egg rejection, suggesting that it may have been subjected to high parasitic history and that egg recognition ability was retained after the loss of parasitism, which needs to be further verified by future experiments.

Fatal mobbing and attack of the common cuckoo by its warbler hosts.

Nest defense is an effective strategy of hosts against parasites. Typically, hosts will aggressively attack brood parasites that approach or visit their nests, which can prevent the parasites from laying eggs or may even lead to the death of the parasites. Few previous studies have specifically reported such fatal cases involving brood parasites and have attributed the cause of death to either drowning or hypothermia after falling into the water following an attack from hosts. In this study, we recorded the process of multiple host individuals of the Oriental reed warbler (Acrocephalus orientalis) mobbing and attacking a female common cuckoo (Cuculus canorus) in the field. We discovered that the immediate cause of the cuckoo's death was the fatal physical damage resulting from the aggressive defense from the hosts, suggesting that frantic pecking and scratching by the hosts is the most proximate cause of mortality among egg-laying female cuckoos. This finding enhances our essential understanding of the effectiveness of host attacks.