Has the importance of the amniote egg been overstated?

The evolution of the amniote egg is commonly regarded as an important milestone in the history of the vertebrates, an innovation that completed the transition from aquatic to fully terrestrial existence by permitting eggs to be laid away from standing water. This view derives ultimately from the recapitulationist theories of Haeckel, and rests on the assumption that extant frogs and salamanders are good models for the reproductive habits of early tetrapods and the ancestors of the amniotes. It also assumes that it is more difficult to lay eggs on land than in water, and that the amniote egg is an adaptation to the physical rigours that eggs encounter in terrestrial environments. Taken together, these assumptions comprise what may be termed the 'Haeckelian framework' for the origin of vertebrate terrestriality. Several independent lines of evidence suggest that the assumptions of the Haeckelian framework are false. There appear to be no theoretical reasons to assume that the evolution of terrestrial egg-laying was difficult, or required a structure as elaborate as the amniote egg. The physical conditions eggs encounter in the terrestrial environments where they are actually laid are quite mild. Land may in fact be an easier place to lay eggs than water. In addition, analysis of the distribution of key reproductive character states among vertebrates provides no evidence that the 'typical amphibian' reproductive mode is primitive for tetrapods. Amniotes are as likely as frogs or salamanders to retain primitive reproductive character states.     

The corn snake yolk sac becomes a solid tissue filled with blood vessels and yolk-rich endodermal cells

The amniote egg was a key innovation in vertebrate evolution because it supports an independent existence in terrestrial environments. The egg is provisioned with yolk, and development depends on the yolk sac for the mobilization of nutrients. We have examined the yolk sac of the corn snake Pantherophis guttatus by the dissection of living eggs. In contrast to the familiar fluid-filled sac of birds, the corn snake yolk sac invades the yolk mass to become a solid tissue. There is extensive proliferation of yolk-filled endodermal cells, which associate with a meshwork of blood vessels. These novel attributes of the yolk sac of corn snakes compared with birds suggest new pathways for the evolution of the amniote egg.     

A new peculiar dinosaur egg,Sankofa pyrenaica oogen. nov. oosp. nov. from the Upper Cretaceous coastal deposits of the Aren Formation,south‐central Pyrenees,Lleida, Catalonia,Spain

Abstract: A new type of small, ovoid dinosaur egg, Sankofa pyrenaica oogen. nov. oosp. nov., with a prismatic type eggshell is described from upper Cretaceous (upper Campanian–Maastrichtian) deposits of the Montsec area, South Pyrenean Central Unit, Lleida, Catalonia, Spain. This egg type was sub‐vertically laid in only two rich monospecific sites of a single stratigraphic layer from coastal deposits of the Aren Formation, interpreted as an emerged beach ridge of a barrier island – lagoon depositional system. The size and shape of these eggs with their asymmetric poles are roughly similar to modern hen eggs, which is unusual in the Cretaceous fossil egg record. Its phylogenetic position clusters with bird and Troodontid eggs. A morphospace analysis of egg shapes shows the similarity of the new egg to a Campanian fossil bird egg from Argentina, both being intermediate between modern‐bird eggs and extinct nonavian theropod eggs. However, the eggshell microstructure of Sankofa pyrenaica differs from that of bird eggs in its incipient squamatic texture. It has a peculiar pattern of interlocking small crystals in the middle of the palisade layer, instead of the thick squamatic structure commonly present in modern avian eggshells. This new egg type is attributed to a small theropod, probably with a single oviduct like birds and whose mosaic distribution of features is a combination between that of birds and nonavian theropods. This enhances the arguments supporting the close phylogenetic relationships between both groups.     

A comparative study of the ultrastructure and mineralogy of calcified land snail eggs (Pulmonata: Stylommatophora)

This study indicates that eggs containing calcium carbonate crystals occur in at least 36 of the 65 known families of the land snails (class Gastropoda: order Stylommatophora). Eggs from 22 of these families were available for examination. The x-ray diffraction data, available for the first time for 21 of these families, shows that these egg shells are all made of calcite only, or of a combination of calcite with smaller amounts of aragonite. All of the snail (body) shells examined were made of aragonite only. This is the first ultrastructural investigation of these egg shells, and it indicates that the eggs exhibit enough structural diversity to allow identification of parental animals to genus, and often to species level solely on the basis of egg shell ultrastructure. All of the calcified eggs may be divided into two groups: (1) partly calcified, with discrete crystals of CaCo₃ dispersed in the jelly layer, and (2) heavily calcified, with a hard, brittle egg shell made of fused crystals of CaCO₃ much like an avian egg. Both types of calcified eggs occur in oviparous as well as in ovoviviparous snails. Because of the wide distribution of calcified eggs in the Stylommatophora, and because of the occurrence of heavily calcified eggs in ancient families such as Partulidae, Endodontidae, and Zonitidae, the calcified egg is viewed as a primitive land snail trait associated with terrestrial adaptation. The function of the calcified egg shell, in addition to mechanical support of egg contents, is to supply the developing embryo with enough calcium to form the embryonic shell by the time of hatching.     

Patterns of reproductive‐mode evolution in Old World tree frogs (Anura,Rhacophoridae)

The Old World tree frogs (Anura: Rhacophoridae), with 387 species, display a remarkable diversity of reproductive modes – aquatic breeding, terrestrial gel nesting, terrestrial foam nesting and terrestrial direct development. The evolution of these modes has until now remained poorly studied in the context of recent phylogenies for the clade. Here, we use newly obtained DNA sequences from three nuclear and two mitochondrial gene fragments, together with previously published sequence data, to generate a well‐resolved phylogeny from which we determine major patterns of reproductive‐mode evolution. We show that basal rhacophorids have fully aquatic eggs and larvae. Bayesian ancestral‐state reconstructions suggest that terrestrial gel‐encapsulated eggs, with early stages of larval development completed within the egg outside of water, are an intermediate stage in the evolution of terrestrial direct development and foam nesting. The ancestral forms of almost all currently recognized genera (except the fully aquatic basal forms) have a high likelihood of being terrestrial gel nesters. Direct development and foam nesting each appear to have evolved at least twice within Rhacophoridae, suggesting that reproductive modes are labile and may arise multiple times independently. Evolution from a fully aquatic reproductive mode to more terrestrial modes (direct development and foam nesting) occurs through intermediate gel nesting ancestral forms. This suggests that gel nesting is not only a possible transitional state for the evolution of terrestriality, but also that it is a versatile reproductive mode that may give rise to other terrestrial reproductive modes. Evolution of foam nesting may have enabled rhacophorids to lay a larger number of eggs in more open and drier habitats, where protection from desiccation is important. Terrestrial direct development allows frogs to lay eggs independent of bodies of water, in a diversity of humid habitats, and may represent a key innovation that facilitated the evolution of nearly half of all known rhacophorid species.     

Development in frogs with large eggs and the origin of amniotes

The origin of the amniote egg is one of the most significant events in the evolution of terrestrial vertebrates. This innovation was probably driven by increased egg size, and to find potential parallels, we can examine the derived development of extant amphibians with large eggs. The embryo of the Puerto Rican tree frog, Eleutherodactylus coqui, exhibits an alteration of its fate map and a secondary coverage of its yolky cells, reflecting the large 3.5 mm egg. Comparable changes may have occurred with the derivation of an amniote pattern of development. Future investigations should focus on the molecular organization of the egg. In the model amphibian for development, Xenopus laevis, information for embryonic germ layers, the dorsal axis, and germ cells is stored mainly as localized RNAs at the vegetal pole of the egg. These localizations would likely be changed with increased egg size. A review of the orthologues of the key X. laevis genes raises the possibility that their activities are not conserved in other vertebrates.     

Differences in egg size, shell thickness, pore density, pore diameter and water vapour conductance between first and second eggs of Snares Penguins Eudyptes robustus and their influence on hatching asynchrony

Brood reduction in birds is frequently induced by hatching asynchrony. Crested penguins (genus Eudyptes) are obligate brood reducers, but in contrast to most other birds, first‐laid eggs are considerably smaller in size than second‐laid eggs; furthermore, first‐laid eggs hatch after their siblings. The mechanisms underlying this reversal in size and hatching order remain unclear. In this study, we tested whether the second‐laid eggs of Snares Penguins Eudyptes robustus have a higher eggshell porosity allowing them to maintain a higher metabolic rate throughout incubation and to hatch before their first‐laid siblings. We investigated differences in egg size, shell thickness, pore density, pore diameter and water vapour conductance between first and second eggs within clutches and examined the influence of these shell characteristics on hatching asynchrony. First‐laid eggs of Snares Penguins were approximately 78% of the size of the larger second eggs. Second‐laid eggs had considerably thicker shells and more pores per cm² than first eggs, whereas pore diameter did not differ between eggs. Water vapour conductance was greater in second‐ (16.8 mg/day/torr) than in first‐laid eggs (14.9 mg/day/torr). The difference in water vapour conductance between first‐ and second‐laid eggs within clutches was related to hatching patterns. In nests where second eggs hatched before first‐laid eggs, second eggs had a considerably greater water conductance than their sibling, whereas in nests where both eggs hatched on the same day, the difference in water conductance between eggs was very small, and in a few nests where small first eggs hatched before their larger sibling, they had a greater water conductance than their larger second‐laid nestmate. Surprisingly few studies have investigated differences in shell characteristics between eggs within clutches and associated effects on hatching asynchrony. This study has demonstrated that such differences exist between eggs within clutches and that they can influence hatching patterns.     

ULTRASTRUCTURAL AND FUNCTIONAL MORPHOLOGY OF EGGSHELLS SUPPORTS THE IDEA THAT DINOSAUR EGGS WERE INCUBATED BURIED IN A SUBSTRATE

Abstract:  The reproductive biology of dinosaurs is of great interest, particularly in light of the many fossil eggs assigned to this group. The ultrastructural characteristics of dinosaur eggshells are examined in order to calculate water vapour conductance, which indicates the nesting environment. Data were mainly derived from the literature but new values are also presented. Allometric analyses were carried out on a variety of shell parameters against predicted egg mass, and comparison was made with allometric equations for bird eggs. Shell thickness was generally larger than seen for extant birds. Total pore number and pores per unit area were similar to values predicted from bird eggs. Total pore area showed an isometric increase with egg mass, parallel to the relationship for birds, but the constant value was an order to magnitude higher than the bird values. Pore radius was unaffected by egg mass. Water vapour conductance showed an allometric increase with egg mass, parallel to the bird values, but for any given egg mass values for dinosaurs were an order of magnitude higher. Mass-specific water vapour conductance was unaffected by egg mass but was an order of magnitude higher than the bird values. Water vapour conductance per pore showed an allometric decrease with egg mass but again the predicted values were an order of magnitude higher than for bird eggs. The ultrastructural characteristics of dinosaur eggshells indicate that the nesting environment had to be saturated with water vapour and that dinosaur eggs had to be fully buried in a substrate. In this sense, therefore, dinosaur eggs resemble more those of modern reptiles than those of birds. As a consequence, maintenance of incubation conditions would have depended on the prevailing environment.     

Prospects of herbivore egg‐killing plant defenses for sustainable crop protection

Due to a growing demand of food production worldwide, new strategies are suggested to allow for sustainable production of food with minimal effects on natural resources. A promising alternative to the application of chemical pesticides is the implementation of crops resistant to insect pests. Plants produce compounds that are harmful to a wide range of attackers, including insect pests; thus, exploitation of their natural defense system can be the key for the development of pest‐resistant crops. Interestingly, some plants possess a unique first line of defense that eliminates the enemy before it becomes destructive: egg‐killing. Insect eggs can trigger (1) direct defenses, mostly including plant cell tissue growth or cell death that lead to eggs desiccating, being crushed or falling off the plant or (2) indirect defenses, plant chemical cues recruiting natural enemies that kill the egg or hatching larvae (parasitoids). The consequences of plant responses to eggs are that insect larvae do not hatch or that they are impeded in development, and damage to the plant is reduced. Here, we provide an overview on the ubiquity and evolutionary history of egg‐killing traits within the plant kingdom including crops. Up to now, little is known on the mechanisms and on the genetic basis of egg‐killing traits. Making use of egg‐killing defense traits in crops is a promising new way to sustainably reduce losses of crop yield. We provide suggestions for new breeding strategies to grow egg‐killing crops and improve biological control.     

The extraembryonic serosa protects the insect egg against desiccation

Insects have been extraordinarily successful in occupying terrestrial habitats, in contrast to their mostly aquatic sister group, the crustaceans. This success is typically attributed to adult traits such as flight, whereas little attention has been paid to adaptation of the egg. An evolutionary novelty of insect eggs is the serosa, an extraembryonic membrane that enfolds the embryo and secretes a cuticle. To experimentally test the protective function of the serosa, we exploit an exceptional possibility to eliminate this membrane by zerknüllt1 RNAi in the beetle Tribolium castaneum. We analyse hatching rates of eggs under a range of humidities and find dramatically decreasing hatching rates with decreasing humidities for serosa-less eggs, but not for control eggs. Furthermore, we show serosal expression of Tc-chitin-synthase1 and demonstrate that its knock-down leads to absence of the serosal cuticle and a reduction in hatching rates at low humidities. These developmental genetic techniques in combination with ecological testing provide experimental evidence for a crucial role of the serosa in desiccation resistance. We propose that the origin of this extraembryonic membrane facilitated the spectacular radiation of insects on land, as did the origin of the amniote egg in the terrestrial invasion of vertebrates.     

Pallisentis rexus from the Chiang Mai Basin, Thailand: ultrastructural studies on egg envelope development and the mechanism of egg expansion

Pallisentis rexus Wongkham & Whitfield, 1999 (Eoacanthocephala: Quadrigyridae) infects the freshwater snakehead fish, Channa striata, in the Chiang Mai Basin, Thailand. All stages of egg development within the body cavity of the female parasite were observed, using transmission electron microscopy. Changes in mature eggs after contact with water were also investigated. The mature egg has five egg envelopes separated from each other by four gaps. The fertilization membrane, which is formed first, is pushed centrifugally by other, subsequently formed, envelopes and gaps, which produces a final total shell thickness of 8-36 microm around the acanthor. The disappearance of the outermost layer and the unpleating of an adjacent inner layer causes the expansion of eggs on contact with water. The volume of an expanded egg is approximately 27 times that of an unexpanded one, but the density of eggs is reduced from a value greater than water to one almost equal to water. This is believed to aid the dispersion of eggs.     

Crocodylomorph eggs and eggshells from the Adamantina Formation (Bauru Group), Upper Cretaceous of Brazil

Abstract: Compared with crocodylomorph body fossils, the record of fossil crocodiloid eggs is scarce and poorly understood, a gap partially attributed to their typically thin eggshell, which is not conducive to preservation. A remarkable new association of well‐preserved eggs and eggshells from the Adamantina Formation (Bauru Group, Upper Cretaceous) is described and compared to other known materials, while the significance of their unique oological features is discussed. These eggs constitute a new ootaxon, Bauruoolithus fragilis oogen. et oosp. nov., diagnosed by the following characteristics: elongate and elliptical egg with blunt ends; length‐to‐diameter ratio of 1:0.55; outer surface slightly undulating; shell thickness ranging from 0.15 to 0.25 mm; pore openings elliptical or teardrop‐shaped, ranging from 30 to 80 μm in diameter; and shell units wider than higher, with the interstices forming an obtuse triangle. Specimens of Bauruoolithus also show only slight signs of extrinsic degradation that, coupled with the evidence that some of them constitute hatched eggs, suggests that the egg‐laying taxon had a different pattern of egg incubation, in which the hatchling could break through the rather thin eggshell relatively easily and that the extrinsic degradation of the eggshell was not necessary. This contrasts with the pattern of incubation for all other known crocodylomorphs and crocodiloid eggs, where extrinsic degradation is a key component of the hatching process.     

Effects of experimental calcium availability,egg parameters and laying order on Great Tit Parus major eggshell pigmentation patterns

Many bird species lay eggs speckled with protoporphyrin‐based spots, however, for most of them the function of eggshell spotting is unknown. A plausible hypothesis is that protoporphyrin might have a structural function in strengthening the eggshell and is therefore deposited when calcium is scarce. In this study, we experimentally provided Great Tit Parus major females with supplemental calcium to examine its effect on the protoporphyrin‐based maculation of their eggs. In addition, we studied variation in eggshell pigmentation patterns in relation to other egg parameters and laying order. Calcium‐supplemented females laid larger eggs but shell thickness was not significantly affected by the treatment. Calcium supplementation may reduce the time and energy females devote to searching for calcium‐rich material, so that they can collect more nutrients and so lay larger eggs. Furthermore, pigment darkness was associated with egg volume and shape, which suggests that female quality and environmental food availability may also influence the shell pigmentation pattern. Within clutches, later‐laid eggs had larger and darker spots that were distributed more unevenly on the shell surface. This within‐clutch pattern could be explained by the increase in egg volume and egg shape and a decline in shell thickness with egg‐laying order, which characteristics were all related to shell‐spotting pattern. Eggs with a coronal ring had thinner shells, but pigment intensity and spot size were not related to shell thickness. Thus, our results suggest that concentrated spotting distribution may have a mechanical function, supporting the structural‐function hypothesis.     

Does maternal care evolve through egg recognition or directed territoriality?

The mechanism that facilitates the evolution of maternal care is ambiguous in egg‐laying terrestrial vertebrates: does the ability of mothers to recognize their own eggs lead them under some circumstances to begin providing care or can maternal care evolve from simply being in close proximity to the eggs (e.g. through territorial behaviour)? This question is difficult to answer because in most species, parental care is either absent altogether or present; in only a few species we have the opportunity to observe intraspecific variation in the expression of parental care. We studied a population of long‐tailed skinks (Eutropis longicaudata) in which females have recently evolved maternal care from a noncaring state. Females on Orchid Island, Taiwan, remain with their eggs during incubation and when doing so, actively deter egg predation by egg‐eating snakes (Oligodon formosanus); in all other populations, females lack post‐ovipositional maternal care. Nest‐guarding females on Orchid Island (i) showed antipredator behaviours only in the original nest site in which they laid eggs, even after we removed all of the eggs or substituted them with those of a conspecific; (ii) protect any eggs present inside the original nest site (even when the eggs belong to a conspecific); and (iii) develop this behaviour while gravid (i.e. prior to laying eggs). This supports the hypothesis that long‐tailed skinks cannot recognize their own eggs, suggesting that maternal care is a directed form of territoriality only expressed towards egg‐eating snakes and only during reproduction. Nest guarding is among the most primitive forms of parental care, and the recent evolution of this behaviour in a single population provides insight into one of the mechanisms by which parental care can originate in terrestrial vertebrates.     

A new Upper Cretaceous titanosaur nesting site from La Rioja (NW Argentina), with implications for titanosaur nesting strategies

Cretaceous titanosaur nesting sites are currently known only from Europe, Asia and South America. In the latter, only the Auca Mahuevo and Sanagasta nesting sites have been confidently assigned to this clade of sauropod dinosaurs. Here we report the discovery of the first eggs and egg clutches found at Tama, a new Upper Cretaceous fossiliferous locality in the Los Llanos Formation, Sierra de Los Llanos (La Rioja, NW Argentina). At least five egg clutches, several partially preserved, isolated eggs and many eggshell fragments were discovered in a single outcrop of a sandstone horizon which represents a cumulative palaeosol profile. Although the mechanical and digital preparation of eggs did not reveal any embryonic remains in ovo, the morphology of the eggs and eggshells closely matches that of titanosaur eggs and eggshells found worldwide. The morphology and spatial grouping of the titanosaur eggs from Tama, along with geological observations support a burrow‐nesting strategy for these dinosaurs. Although the Sanagasta and Tama eggs were found in the same stratigraphical unit and share several morphological characters, they clearly differ in shell thickness and egg size. This, coupled with the interpretation of different sedimentary contexts for these nesting sites, strongly suggests that at least two different titanosaur species nested in La Rioja during the Late Cretaceous, using different nesting strategies. The occurrence of this new titanosaur nesting site in a semiarid palaeoenvironment represents an interesting case study for the reproductive biology of the titanosaur dinosaurs, particularly their labile nesting behaviour.     

Abbreviation of larval development and extension of brood care as key features of the evolution of freshwater Decapoda

The transition from marine to freshwater habitats is one of the major steps in the evolution of life. In the decapod crustaceans, four groups have colonized fresh water at different geological times since the Triassic, the freshwater shrimps, freshwater crayfish, freshwater crabs and freshwater anomurans. Some families have even colonized terrestrial habitats via the freshwater route or directly via the sea shore. Since none of these taxa has ever reinvaded its environment of origin the Decapoda appear particularly suitable to investigate life‐history adaptations to fresh water. Evolutionary comparison of marine, freshwater and terrestrial decapods suggests that the reduction of egg number, abbreviation of larval development, extension of brood care and lecithotrophy of the first posthatching life stages are key adaptations to fresh water. Marine decapods usually have high numbers of small eggs and develop through a prolonged planktonic larval cycle, whereas the production of small numbers of large eggs, direct development and extended brood care until the juvenile stage is the rule in freshwater crayfish, primary freshwater crabs and aeglid anomurans. The amphidromous freshwater shrimp and freshwater crab species and all terrestrial decapods that invaded land via the sea shore have retained ocean‐type planktonic development. Abbreviation of larval development and extension of brood care are interpreted as adaptations to the particularly strong variations of hydrodynamic parameters, physico‐chemical factors and phytoplankton availability in freshwater habitats. These life‐history changes increase fitness of the offspring and are obviously favoured by natural selection, explaining their multiple origins in fresh water. There is no evidence for their early evolution in the marine ancestors of the extant freshwater groups and a preadaptive role for the conquest of fresh water. The costs of the shift from relative r‐ to K‐strategy in freshwater decapods are traded‐off against fecundity, future reproduction and growth of females and perhaps against size of species but not against longevity of species. Direct development and extension of brood care is associated with the reduction of dispersal and gene flow among populations, which may explain the high degree of speciation and endemism in directly developing freshwater decapods. Direct development and extended brood care also favour the evolution of social systems, which in freshwater decapods range from simple subsocial organization to eusociality. Hermaphroditism and parthenogenesis, which have evolved in some terrestrial crayfish burrowers and invasive open water crayfish, respectively, may enable populations to adapt to restrictive or new environments by spatio‐temporal alteration of their socio‐ecological characteristics. Under conditions of rapid habitat loss, environmental pollution and global warming, the reduced dispersal ability of direct developers may turn into a severe disadvantage, posing a higher threat of extinction to freshwater crayfish, primary freshwater crabs, aeglids and landlocked freshwater shrimps as compared to amphidromous freshwater shrimps and secondary freshwater crabs.     

Experimentally induced effects of water temperature and immersion time on reproductive output of bivalves in the Wadden Sea

Results of experiments are reported on the effects of water temperature and immersion time in winter on egg size and egg numbers in three intertidally living bivalves in the Dutch Wadden Sea, the Baltic tellin Macoma balthica, the common cockle Cerastoderma edule and the common mussel Mytilus edulis. Macoma (14–17 mm shell length) produced large eggs (diameter of 107 μm) in relatively small numbers (20 000–70 000) in early spring. Later in spring, Cerastoderma (28–33 mm shell length) produced smaller eggs (77 μm, excluding the surrounding jelly layer) in tenfold larger numbers (200 000–700 000). Mytilus (45–55 mm shell length) spawned even smaller eggs (72 μm) in high (but not easily assessed) numbers over a more extended period. In Macoma egg size was not affected by winter temperatures or immersion time. Effects of winter–spring temperatures and immersion time on egg size could be demonstrated in Cerastoderma. Smaller eggs were produced at the higher temperatures. Effects of immersion time were non-consistent: at lower water temperatures larger, but at higher temperatures smaller eggs were produced by animals kept at longer immersion times. In Mytilus, no temperature effects were observed. However, a longer immersion time resulted in larger eggs. In Macoma as well as in Cerastoderma significantly more eggs were produced at the lower temperature. Immersion time effects were most pronounced at the lower temperature, where more eggs were produced at the subtidal level than at the tidal level. At the higher water temperature differences between egg numbers produced at the two tidal levels were small. Just prior to spawning, egg numbers were strongly positively related to body mass at a certain shell length.     

The effect of embryonic development on metal and calcium content in eggs and eggshells in a small passerine

Published information relating to changes in the chemical element content of avian eggs caused by embryonic development is extremely scarce, although it may be crucial for understanding both the presence of anthropogenic pollutants as well as physiological levels of micronutrients. We assessed the variation in concentrations of calcium (Ca) and magnesium (Mg) and nine trace elements: seven essential (chromium (Cr), copper (Cu), nickel (Ni), manganese (Mn), iron (Fe), cobalt (Co) and zinc (Zn)) and two non‐essential (lead (Pb) and cadmium (Cd)) in shells and contents (both egg yolk and egg white) of embryonated and non‐embryonated eggs. We investigated the eggs of the Eurasian Reed Warbler Acrocephalus scirpaceus, a large proportion of whose eggs are infertile in our study population (almost 43% of clutches contain unhatched eggs) as well as significant embryo‐induced eggshell thinning at the equator of embryonated eggs. We found significantly higher concentrations (≥ 22.7%) of all the focal elements in the contents of embryonated eggs in comparison with non‐embryonated eggs, and a very pronounced one for Ca (nearly twice as high). The shells of embryonated eggs contained significantly higher concentrations of Zn (104.1%), Fe (56.5%), Pb (32.8%) and Cu (28.0%) but significantly lower ones of Co (8.9%) and Ca (9.3%) than the shells of non‐embryonated eggs. The simultaneous higher concentrations of all elements in the content of thinner‐shelled embryonated eggs suggest the parallel transfer of these elements along with Ca resorption from the shell into the egg interior during embryo formation. The higher concentration of most elements in the thinner shells of embryonated eggs may be indicative of the maternal deposition of some of these elements in a shell layer not subject to embryonic depletion, or in the eggshell membrane. Our results highlight the need for the careful selection of egg samples, which should differentiate between embryonated and non‐embryonated eggs in the analytical treatment of eggs and eggshells.     

The evolution of host‐specific variation in cuckoo eggshell strength

Cuckoo eggs are renowned for their mimicry of different host species, leading to the evolution of host‐specific races (or ‘gentes’) defined by egg colour and pattern. This study aims to test the prediction that another property of parasitic eggs, namely shell strength, might also have experienced divergent selection within cuckoo species. Host races of the common cuckoo Cuculus canorus encountering stronger host rejection have thicker‐shelled eggs than those parasitising less discriminating species, as expected if egg strengthening discourages host rejection. Moreover, in the diederik cuckoo Chrysococcyx caprius, eggshell thickness was correlated across cuckoo gentes and host species, as expected if eggshell strength has been involved in coevolutionary interactions. This is the first report of host‐specific differences in cuckoo egg properties other than colour and pattern and lends correlational support to the hypothesis that the strong eggshells of brood parasites are an adaptation to reduce host rejection.     

Three new species of Huffmanela Moravec, 1987 (Nematoda: Trichosomoididae) from the gills of marine fish off New Caledonia

Three new species of Huffmanela, here described from eggs only, are reported from the gills of marine fish caught off Nouméa, New Caledonia. Eggs of Huffmanela branchialis n. sp., from Nemipterus furcosus (Nemipteridae), are 45-52 (mean 48) microm in length and 23-30 (mean 25) microm in width, with thin shells. Each egg is enclosed in a thin membrane forming a spindle-shaped envelope 53-85 (mean 63) microm in length. Eggs of H. filamentosa n. sp., from Gymnocranius grandoculis (Lethrinidae), are 48-53 (mean 50) microm in length and 25-30 (mean 27) microm in width, with thin shells. Each egg bears a few long (150 microm), thin filaments. Eggs of these two new species were compared to those of H. paronai Moravec & Garibaldi, 2000, which are redescribed. Eggs of H. ossicola n. sp. were found within the branchial arch bone of Bodianus loxozonus (Labridae) and also filled the spinal chord bone and other bones. This is the first species of Huffmanela reported from bone tissue. Eggs are large, 72-88 (mean 79) microm in length and 32-40 (mean 36) microm in width, with a very thick shell. Each egg is covered with numerous filaments enclosed in a thin envelope. Fresh eggs were unembryonated, but embryos were visible after incubation in seawater. The three new species can be distinguished from other species of Huffmanela by size and the nature of the egg covering. Egg morphology of and their location in the host suggest different life-cycles: those of the first two species (small eggs, thin shells, egg covering possibly favouring flotation) are released from the gill mucosa with the turnover of living tissues and immediately continue their life-cycle, but eggs of H. ossicola (large eggs, thick shell) are only available for the continuation of the life-cycle after the host's death.