Comparative study on ovarian structures in scorpaenids: possible evolutional process of reproductive mode

The reproductive modes of the Scorpaenidae are extremely varied: oviparity, viviparity, and even spawning of internally fertilized eggs or embryos (zygoparity or embryoparity), as in Helicolenus, are known. The ovarian structure of this family is divided into two types by the arrangement of the stroma and the ovarian cavity. One type is the ovary in which the lamella-like stroma develops from the ovarian hilus located on the dorsal side and where the ovarian cavity is located on the ventral side of ovary, classified as “cystovarian type II-1” by Takano (1989). In the other type, the stroma in the ovary develops radially around the blood circulatory system that traverses the center of the ovary, and then the ovarian cavity surrounds all the ovary, classified as “cystovarian type II-3” by Takano (1989). In the present analysis, previous reports about ovarian structure and the relationship to the reproductive mode of scorpaenids were described, and the ovarian structure of eight genera of Scorpaenidae was examined. The ovary of cystovarian type II-1 is seen only in viviparous genera and is not seen in oviparous genera. However, the cystovarian type II-1 is a general structure in other families of Scorpaeniformes, and this structure could be considered a primitive type of ovary rather than that acquired by the process of evolution from oviparity to viviparity. The ovary of cystovarian type II-3 is seen in all six oviparous genera and the one zygoparous genus examined. The ovary of this type is not found in any other family of teleosts, so it could be a structure originally divided in Scorpaenidae. In the genera having the cystovarian type II-3 ovary, there is a common feature of spawning: a floating egg mass encompassed by the gelatinous material. We postulate that the evolution of reproductive mode in the scorpaenid fishes is as follows: Sebastes and Sebastiscus have a primitive ovary in which viviparity has developed, whereas the genera that spawn a floating egg mass evolved the ovarian structure from primitive type to cystovarian type II-3, and further zygoparity, such as in Helicolenus, evolved from them.     

REPTILIAN VIVIPARITY AND DOLLO'S LAW

It has been suggested repeatedly that the evolutionary transition from oviparity (egg-laying) to viviparity (live-bearing) in reptiles is irreversible. However, these adaptive arguments have yet to be tested by detailed examination of the phylogenetic distribution of oviparity and viviparity across a broad range of taxa. Using available data on reproductive modes and phylogenetic relationships within reptiles, we here quantify the numbers and directions of evolutionary transitions between oviparity and viviparity. Phylogenetic relationships among three diverse squamate groups (scincid lizards, colubrid snakes, elapid snakes) are currently inadequately known for inclusion in this study Among the remaining reptiles, oviparity has given rise to viviparity at least 35 times. Five possible instances of reversals (from viviparity to oviparity) are identified, but closer examination indicates that all have weak empirical support (i.e., they could be “unreversed” with little loss in parsimony, and/or are based on poorly substantiated phylogenetic hypotheses). Viviparity is clearly more frequently (and presumably easily) gained than lost in several disparate groups so far examined (reptiles, fishes, polychaete worms); this evolutionary bias should be considered when reproductive mode is optimized on a phylogeny or employed in phylogenetic reconstruction.     

THE EVOLUTION OF OVIPARITY WITH EGG GUARDING AND VIVIPARITY IN LIZARDS AND SNAKES: A PHYLOGENETIC ANALYSIS

This paper investigates the evolution of viviparity and of egg guarding in lizards and snakes in which three modes of reproduction can be described: oviparity without egg guarding, oviparity with egg guarding, and viviparity. All possible transitions of reproductive modes were detected in each taxon using Maddison's method. We then tested two specific hypotheses. First, egg guarding can be regarded as an alternative to viviparity. A relatively frequent association of egg guarding and viviparous species in the same taxon may be due to similar environmental conditions or species characteristics leading to two different solutions. Second, egg guarding may facilitate the evolution of viviparity. This hypothesis is supported by the high frequency of viviparous species in taxa containing egg guarding species and by a tendency for prolonged uterine retention of eggs in brooding squamates. Our analyses demonstrate that the first hypothesis is the best supported. Egg guarding and viviparity most often evolved independently. If a major benefit of egg guarding is the repulsion of potential predators, size is one of the most obvious morphological characters that should be correlated with the evolution of reproductive modes. The two reproductive traits were correlated to a reduction in body size for viviparous species and an increase in body size for egg guarding species. This could partly explain why the evolution of these reproductive modes seems almost antagonist.     

MITOCHONDRIAL DNA SEQUENCE-BASED PHYLOGENY AND THE EVOLUTION OF VIVIPARITY IN THE SCELOPORUS SCALARIS GROUP (REPTILIA,SQUAMATA)

The lizard genus Sceloporus contains both oviparous and viviparous species. The scalaris complex is the only monophyletic group within the genus that includes both reproductive modes, thus it is particularly well suited for studies of the evolution of viviparity. Approximately 874 nucleotides of mtDNA sequence data, collected from 38 specimens, comprising 25 populations of all five recognized species within the group, were used in a phylogenetic analysis of the origin of viviparity. Viviparity appears to have evolved twice in this group: once in S. goldmani, included in a clade formed by a northern group consisting of S. scalaris, S. chaneyi, and S. goldmani, and one more time in S. bicanthalis, included in the southern group formed by S. bicanthalis and S. aeneus. An oviparous population of S. bicanthalis nested within that viviparous clade, indicates that reversal from viviparity to oviparity may be possible. Degree of sequence divergence among several S. bicanthalis individuals pertaining to a population in which both parity modes occur, was no larger between oviparous and viviparous lizards than among viviparous lizards. This suggests that this population is a single species, and it may represent a transition from oviparity to viviparity or vice-versa.     

Copulation and egg retention in an oviparous Caecilian (Amphibia: Gymnophiona)

Viviparity (i.e., the bearing of live young) has evolved from oviparity (egg laying) independently in various major vertebrate lineages, and several transitional stages have been described. The transition from oviparity to viviparity requires the retention of fertilised eggs in the female reproductive tract. Caecilian amphibians (Gymnophiona) display a considerable diversity of reproductive modes, including oviparity and viviparity. Among amphibians, caecilians have also modified the process of internal fertilisation through a special intromittent organ, or phallus, in males. Here we report the oviposition of “embryonated” eggs ranging from various gastrula-to-neurula stages by female Ichthyophis cf. kohtaoensis (Ichthyophiidae) from North-eastern Thailand. In addition, we describe a copulation resulting in an oviposition of embryonated eggs. Our findings will have implications for the further understanding of the evolutionary reproductive biology of amphibians.     

Thermoregulation during gravidity in the children's python (Antaresia childreni): a test of the preadaptation hypothesis for maternal thermophily in snakes

Pregnant squamate reptiles (i.e. lizards and snakes) often maintain higher and more stable body temperatures than their nonpregnant conspecifics, and this maternal thermophily enhances developmental rate and can lead to increased offspring quality. However, it is unclear when this behaviour evolved relative to the evolution of viviparity. A preadaptation hypothesis suggests that maternal thermophily was a preadaptation to viviparity. Oviparous squamates are unique among oviparous reptiles for generally retaining their eggs until the embryos achieve one fourth of their development. As a result, maternal thermophily by gravid squamates may provide the same thermoregulatory benefits, at least during early development, that have been associated with viviparity. Thus, the evolution of viviparity in squamates may reflect an expanded duration of a pre-existing maternal thermoregulatory behaviour. Despite its evolutionary relevance, thermoregulation during gravidity in oviparous squamates has not yet been explored in depth. In the present study, we examined whether gravidity was associated with thermoregulatory changes in the oviparous children's python, Antaresia childreni . First, we discovered that, compared to most snakes, A. childreni is at an advanced stage of embryonic development at oviposition. Second, using surgically implanted temperature loggers, we detected a significant influence of reproductive status on thermoregulation. Reproductive females maintained higher and less variable body temperatures than nonreproductive females and this difference was most pronounced during the last 3 weeks of gravidity. Overall, these results highlight the continuum between oviparity and viviparity in squamate reptiles and emphasize the importance of thermal control of early embryonic development independent of reproductive mode.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 499–508.     

Biological observations on the bristly catshark Bythaelurus hispidus from deep waters off the south‐west coast of India

Biological data are presented for the poorly known bristly catshark Bythaelurus hispidus based on specimens collected from the by‐catch of the commercial deep‐sea shrimp trawl fishery operating in the Arabian Sea at depths of 200–500 m off the south‐west coast of India. One hundred and sixty‐two individuals, which ranged from 120 to 366 mm total length (L_T), were collected for this study. Size‐at‐maturity (L₅₀) for females and males was estimated at 252 and 235 mm L_T, respectively. The reproductive mode of B. hispidus was aplacental viviparity, which is the rarest reproductive mode within the Scyliorhinidae and is considered to be the most advanced of the three reproductive modes occurring within this family. Dietary analysis of stomach contents revealed B. hispidus feeds on a variety of prey, primarily fishes.     

Reproduction and development ofSebastes in the context of the evolution of piscine viviparity

Synopsis Selected aspects of the reproduction and development ofSebastes and other rockfishes are reviewed in the context of piscine viviparity. Among the eight subfamilies of the Scorpaenidae, viviparity is confined to the subfamily Sebastinae; gestation is lumenal and the embryos usually develop to term within the egg envelope. Transitional states from oviparity to viviparity are evident in different species within the family. A scenario for the evolutionary origin of viviparity in rockfishes is derived from an analysis of scorpaeniform reproductive biology. Although viviparity is best developed in the genusSebastes, it is still in a primitive, unspecialized state. Rockfish viviparity is essentially lecithotrophic, i.e. embryonic nutrition is dependent on the energy reserves laid down during oogenesis. In other groups of viviparous fishes, lecithotrophy has been shown to be better suited energetically to seasonally unpredictable habitats, whereas matrotrophy requires a predictable food supply. During the evolution of an essentially primitive form of lecithotrophic viviparity in rockfishes, the advantages of high fecundity associated with oviparity were retained while an enormous increase in the survival rate of the developing embryos was acquired. The basic lecithotrophic pattern of oviparous development was not changed since it offered selective advantages both in terms of energetics and as a basis for retaining a large brood size.     

DID EGG‐LAYING BOAS BREAK DOLLO'S LAW? PHYLOGENETIC EVIDENCE FOR REVERSAL TO OVIPARITY IN SAND BOAS (ERYX: BOIDAE)

Re-evolution of lost complex morphological characters has been proposed for several characters, including insect wings, limbs, eyes in snakes, and digits in lizards, among others. There has also been much interest in whether the transition from oviparity to viviparity is reversible, particularly in squamate reptiles where the transition to viviparity has occurred more times than in any other lineage. Here, we present a phylogenetic analysis of boid snakes based on a concatenated multigene study of all genera of erycines, New and Old World boines, plus other groups thought to be closely related with boines such as monotypic species Calabaria and Casarea . We reconstruct ancestral parity mode on this phylogeny and present statistical evidence that oviparity reevolved in a species of Old World sand boa in the genus Eryx nearly 60 million years after the initial boid transition to viviparity. Remarkably, like other viviparous boas hatchlings of oviparous Eryx lack an egg-tooth providing independent evidence that oviparity is a derived state in these species.     

Transition to viviparity in the order Scorpaeniformes: Brief review

Among teleost percoid fishes (Percomorpha), the subordes Scorpaenoidei (Scorpaeniformes) and Zoarcoidei (Perciformes) are of particular interest due to the presence of intermediate stages between oviparity and viviparity in several species. We analyze the features of reproductive biology in the fishes of the suborder Scorpaenoidei, including morphology of the urogenital system, gametogenesis, gamete ultrastructure, and embryonic development. The new materials are represented for three tropical coral reef fish species with external insemination from the family Scorpaenidae (Scorpaenopsis possi, Sebastapistes cyanostigma, and Dendrochirus zebra). In particular, a hypertrophied urinary bladder used for storage of mature sperm for insemination of pelagic egg clutch is described in the males. Evolutionary transition to viviparity within the family Sebastidae is discussed. Alternative reproductive strategies are registered in viviparous fishes from the suborders Scorpaenoidei and Zoarcoidei. They are connected with the production of a large number of morphologically weakly developed larvae and a small number of individuals at the juvenile state, respectively.     

Inferring the ancestral sexuality and reproductive condition in sponges (Porifera)

Considerable diversity abounds among sponges with respect to reproductive and developmental biology. Their ancestral sexual mode (gonochorism vs. hermaphroditism) and reproductive condition (oviparity vs. viviparity) however remain unclear, and these traits appear to have undergone correlated evolution in the phylum. To infer ancestral traits and investigate this putative correlation, we used DNA sequence data from two loci (18S ribosomal RNA and cytochrome c oxidase subunit I) to explore the phylogenetic relationships of 62 sponges whose reproductive traits have been previously documented. Although the inferred tree topologies, using the limited data available, favoured paraphyly of sponges, we also investigated ancestral character‐state reconstruction on a phylogeny with constrained sponge monophyly. Both parsimony‐ and likelihood‐based ancestral state reconstructions indicate that viviparity (brooding) was the likely reproductive mode of the ancestral sponge. Hermaphroditism is favoured over gonochorism as the sexual condition of the sponge ancestor under parsimony, but the reconstruction is ambiguous under likelihood, rendering the ancestry of sexuality unresolved in our study. These results are insensitive to the constraint of sponge monophyly when tracing the reproductive characters using parsimony methods. However, the maximum likelihood analysis of the monophyletic hypothetical tree rendered gonochorism as ancestral for the phylum. A test of trait correlation unambiguously favours the concerted evolution of sexuality and reproductive mode in sponges (hermaphroditism/viviparity, gonochorism/oviparity). Although testing ecological hypotheses for the pattern of sponge reproduction is beyond the scope of our analyses, we postulate that certain physiological constrains might be key causes for the correlation of reproductive characters.     

Superfoetative viviparity in a Carboniferous chondrichthyan and reproduction in early gnathostomes

Chondrichthyan fishes have an evolutionary history spanning over 400 million years and are characterized, in part, by internal fertilization. Traditionally, oviparity has been assumed to be the primitive birthing mode for these fishes and for vertebrates in general, with viviparity and matrotrophic nutrition being derived. The fossilized remains of two specimens of Harpagofututor volsellorhinus from the Upper Mississippian of Montana now provide the first direct evidence of matrotrophic live birth in a Palaeozoic chondrichthyan and of superfoetation in an extinct fish. Each female exhibits multiple foetuses of two size groups, indicating simultaneous gestation of multiple litters. There is no evidence of yolk sacs, only preserved organic pigments enveloping the young, suggesting matrotrophically derived material. Young were born large, as head lengths of the largest embryos measured up to 66 per cent of the mother's head length. Comparison of in utero embryos to isolated specimens suggests, unlike all extant chondrichthyans, the absence of a juvenile stage and rapid maturity. These new data suggest the advantages of superfoetative viviparity for a small bodied fish in a 318 Myr old species‐ and predator‐rich marine bay. In the greater view of gnathostome evolution, this finding combines with other recent discoveries to document that multiple, and not necessarily closely related, species of both placoderms and chondrichthyans exhibited viviparity by the Upper Devonian and the Upper Mississippian. The capacity for internal fertilization probably predisposed members of these lineages to develop viviparity so early in gnathostome history. Yet, the surprising range of viviparity exhibited at this stage of vertebrate evolution emphasizes that derived reproductive strategies had evolved in gnathostomes by 380–318 million years ago. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161 , 587–594.     

Reproduction of the blacktip sawtail catshark,Galeus sauteri,in the waters off northeastern Taiwan

The reproductive biology of blacktip sawtail catsharks,Galeus sauteri, in northeastern Taiwan waters was investigated. Male catsharks possessed paired testes producing spermatozoa, which were then stored in the epididymides all year round. No spermatophores were observed in the lower ductus deferens. Only the right ovary was functional in females, oogonia being formed in the ovarian cortex and then developing into mature ova. Ova exceeding 17 mm in diameter were ovulated. Pregnant individuals contained one eggcase only, in each side of the uterus. No hatched embryos were observed in such eggcases. The size at which 50% of blacktip sawtail catshark specimens were mature was 410–420 mm and 350–360 mm for females and males, respectively. The species does not have a well-defined reproductive season.     

Brood care among basommatophorans: a unique reproductive strategy in the freshwater limpet snail Protancylus (Heterobranchia: Protancylidae), endemic to ancient lakes on Sulawesi, Indonesia

In molluscan taxa inhabiting marine environments oviparity and reproduction via planktonic larvae is predominant while incubation and viviparity is most frequently found in taxa inhabiting brackish or freshwater aquatic habitats. Brooding has evolved repeatedly and independently in several limnic taxa among Bivalvia and Gastropoda. However, among basommatophoran gastropods no such cases were yet known. We here report on a unique reproductive strategy involving brood care in the lacustrine freshwater limpet genus Protancylus, endemic to the ancient lakes on central Sulawesi (former Celebes), Indonesia, namely the Lake Poso and the Malili lake system, because this constitutes the first known case of this behaviour among the Basommatophora. Protancylus live exclusively as epizoans on those pachychilid gastropods of the viviparous genus Tylomelania, also a Sulawesi endemic species, that inhabit mostly soft substrates. We found that the two known species Protancylus pileolus from Lake Poso and P. adhaerens from the Malili lake system both retain gelatinous egg strings underneath their outer mantle, where up to 15 (mostly eight or nine) shelled juveniles are brooded. Nourishment is provided within the egg capsule only. Thus, brood care in Protancylus resembles the reproductive strategy found recently among pachychilid gastropods Jagora from the Philippines, but differs from euviviparous (i.e. matrotrophic) incubation among thiarid gastropods possessing a brood pouch with juveniles being nourished via a ‘pseudoplacenta’ in several taxa.     

Lerista bougainvillii,a case study for the evolution of viviparity in reptiles

Many factors, both environmental and biotic, have been suggested to facilitate or hinder the evolution of viviparity (live-bearing) in reptiles. Viviparity has evolved recently within the Australian scincid lizard Lerista bougainvillii and the species includes oviparous, viviparous, and reproductively intermediate (with prolonged egg retention) populations; thus, it offers an exceptional opportunity to evaluate the validity of these hypotheses. We carried out such tests by (i) comparing environmental conditions over the geographic ranges occupied by oviparous, viviparous, and intermediate populations (to identify possible selective forces for the evolution of viviparity), and (ii) comparing morphological, reproductive and ecological traits of L. bougainvillii with those of other sympatric scincid species (to identify traits that may have predisposed this taxon to the evolution of viviparity). The areas occupied by viviparous L. bougainvillii are significantly colder than those occupied by both their intermediate and oviparous conspecifics, in accord with the “cold-climate” hypothesis for reptilian viviparity. Rainfall is similar over the ranges of the three forms. Climatic unpredictability (as assessed by the magnitude of year-to-year thermal variation) is lower for viviparous animals, in contradiction to published speculations. Comparison with 31 sympatric scincid species showed that L. bougainvillii is not atypical for most of the traits we measured (e.g., body size, clutch size, thermal preferenda and tolerances). However, oviparous L. bougainvillii do display several traits that have been suggested to facilitate the evolution of viviparity. For example, pregnancy does not reduce locomotor ability of females; the lizards are semi-fossorial; even the oviparous females produce only a single clutch of eggs per year; and they ovulate relatively late in summer, so that the time available for incubation is limited.     

Probable evolution of the coelacanth's reproductive style: lecithotrophy and orally feeding embryos in cichlid fishes and in Latimeria chalumnae

Synopsis The living coelacanth is a livebearer. Yolk seems to be the main source of nutrients and of oxygen to the embryo (fetus). Long before birth, young may also possibly feed orally on histotrophe secretion and egg debris. This type of reproduction evolved, as in most other fishes, from oviparity. The Carboniferous coelacanth Rhabdoderma exiguum had eggs of much lesser yolk volume and may represent an earlier form of oviparity with hiding, guarding or brooding type of parental care. The Jurassic coelacanth Holophagus (Undina) and the Cretaceous Axelrodichthys appear to have already evolved the internal-bearing style. Much of this evolutionary sequence is similar to that in cichlids. Ancestral cichlids are substrate tenders and nesters, with small eggs, little yolk and a feeding larva with indirect development. Mouthbrooding cichlids evolved a few, large eggs with denser yolk, direct development and, ultimately, orally feeding embryos while yolk is still in ample supply. Mixed feeding from yolk and orally ingested food in cichlids and in coelacanths is shown to be an enhanced mode of food delivery to the embryos over that from each source separately, in order to produce directly a better developed or larger young at the time of release, i.e. independence. Increase in egg size is regarded as an environmentally induced, altered pattern of yolk synthesis and an initial component of the epigenetic mechanism leading towards greater specialization. Carotenoids are incorporated within the yolk to assist the oxidative metabolism of the developing embryo.     

Mating and fecundity of Dermatophagoides farinae

Studies of the life cycle of cultured Dermatophagoides farinae found that after an initial mating D. farinae females lived for 63.3 ± 64.6 (SD) dafter their egg production period ended .The long period after cessation of egg production for D. farinae suggested D. farinae females could mate multiple times and produce eggs continuously for a longer period. The purpose of this study was to determine if female D. farinae could mate at least two times, and subsequently increase the production of viable eggs over a longer period of time compared to a single mating. Female D. farinae were randomly selected from thriving cultures and isolated in cages. When the females had ceased to lay eggs a male was added to the cage. Fifty-seven percent of the isolated females mated again and produced a second batch of viable eggs. In natural or culture populations, females have continuous availability of males. Therefore, in another experiment, females that emerged from the tritonymphal stages were continuously exposed to fresh males and fecundity and lengths of the reproductive and post reproductive periods were determined. These females had a 11 d longer reproductive period and produced 30.7% more eggs compared to females that only mated one time after they emerged from the tritonymphal stage. However, the post reproductive period was still long (58.6 ± 11.4 [SE] d) the significance of which is not clear. In conclusion, this study revealed that D. farinae females are capable of more than one successful mating that results in increased egg production compared to that of a single mating. It is likely that females mate multiple times in natural and culture populations. It was observed that females actively attract males during the reproductive period but not afterward even though she continues to live a long time. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ontogenesis of the cricket Gryllus argentinus Sauss. (Orthoptera, Gryllidae)

The development of Gryllus argentinus Sauss. was studied under stable laboratory conditions: the temperature of 26°C, the air humidity of 60%, and the photoperiod of 12h light: 12 h dark. The life cycle of Gryllus argentinus includes four stages: egg, pronymph, nymph, and adult. The duration of embryonic development is 18 days. The depth of egg bedding in the peat is 9.63 ± 0.12 mm (n =145), the clutch containing 2–4 eggs. A female can lay over 1100 viable eggs during the entire oviposition period. Nymphal development includes 9 instars and lasts 97 days. The duration of nymphal instars (days) is: I—5; II—6; III—6; IV—6; V—8; VI—10; VII—13; VIII—14; IX—29. The duration of the adult life is 51 days in males and 69 days, in females. In the imaginal ontogenesis of males and females, three periods can be distinguished: pre-reproductive, reproductive, and postreproductive. Males start to emit the aggressive signal on the 6th (5–8th) day (the pre-reproductive period). They enter the reproductive period (start to emit the calling song) on the 9th (8–13th) day. Females enter the reproductive period (become capable of responding to the calling song and of copulation) on the 9th (8–10th) day. Oviposition starts on the day after the first copulation. The reproductive period lasts about 40 (15–59) days in males and 58 (21–70) days in females. The post-reproductive period starts in females at the moment of finishing the egg laying period and in males, with disappearance of reproductive behavior. The period ends in the animal’s death.     

Thermal and reproductive biology of high and low elevation populations of the lizard Sceloporus scalaris: implications for the evolution of viviparity

Viviparity in squamate reptiles is presumed to evolve in cold climates by selection for increasingly longer periods of egg retention. Longer periods of egg retention may require modifications to other reproductive features associated with the evolution of viviparity, including a reduction in eggshell thickness and clutch size. Field studies on the thermal and reproductive biology of high (HE) and low (LE) elevation populations of the oviparous lizard, Sceloporus scalaris, support these expectations. Both day and night-time temperatures at the HE site were considerably cooler than at the LE site, and the activity period was 2 h shorter at the HE than at the LE site. The median body temperature of active HE females was 2°C lower than that of LE females. HE females initiated reproduction earlier in the spring than LE females, apparently in order to compensate for relatively low temperatures during gestation. HE females retained eggs for about 20 days longer than LE females, which was reflected by differences in the degree of embryonic development at the time of oviposition (stages 35.5–37.0 versus stages 31.0–33.5, respectively). These results support the hypotheses that evolution of viviparity is a gradual process, and is favored in cold climates. Females in the HE population exhibited other traits consistent with presumed intermediate stages in the evolution of viviparity; mean eggshell thickness of HE eggs (19.3 m) was significantly thinner than that of LE eggs (26.6 m) and the size-adjusted clutch sizes of HE females (9.4) were smaller than those of LE females (11.2).