Calcite and aragonite in the egg shell of Chelonia mydas L.

The structure of the calcified layer of egg shells from farm-reared and wild turtles has been examined using scanning electron microscopy and infrared analysis. Farm-reared egg shells contained discrete morphologically distinct regions of blocks of calcite and spherulites of aragonite. In contrast, the egg shells from feral populations consisted only of the spherulites. Differences in the impurity contents of the calcite and aragonite structures have been observed. SEM revealed a thin cuticular membrane.     

Studies on the egg shell (oviducal and oviposited) of Chelonia mydas L.

The outer calcified surface of the turtle egg shell consists primarily of crystalline aggregates of calcium carbonate in its aragonite form, together with a small amount (< 5 %) of calcitic material. The latter is first deposited to be followed by aragonite deposition.In the first instance, calcification occurs on the rims of discrete pits formed by the lateral deflection of the ends of soft shell membrane fibres. As crystal deposition continues these pits become filled in and eventually occluded.Micro- and X-ray diffraction analyses of the calcified layer indicate the presence of phosphorus and sulphur. The effects of these elements on the type of crystal deposited, (i.e., aragonite or calcite) is discussed.     

Oviposition Behavior and Offspring Emergence Patterns in Succinea thaanumi, an Endemic Hawaiian Land Snail

In this field study, newly laid egg clutches of Succinea thaanumi, an endemic Hawaiian land snail, were labeled and observed weekly over a 12‐mo period. Snails usually laid their clutches under leaf tips and, moreover, this placement affected snail emergence by mitigating the effects of dehydration through the leaf‐drip phenomenon. During droughts, the gel surrounding egg clutches dehydrated, pasting the embryos to the backs of leaves. The hygroscopic gel rehydrated once rains resumed. The majority of newly laid clutches contained shell‐less embryos which required more time for emergence than embryos laid with visible shells. More shelled embryos were laid subsequent to a 3‐wk drought, indicating that snails retained their eggs during adverse environmental conditions and laid them when conditions improved. The snails’ abilities to retain eggs in adverse conditions, but to lay more unshelled embryos over their lifetimes, contributed to their overall reproductive success.     

Aragonite shells are more ancient than calcite ones in bivalves: new evidence based on omics

Two calcium carbonate crystal polymorphs, aragonite and calcite, are the main inorganic components of mollusk shells. Some fossil evidences suggest that aragonite shell is more ancient than calcite shell for the Bivalvia. But, the molecular biology evidence for the above deduction is absent. In this study, we searched for homologs of bivalve aragonite-related and calcite-related shell proteins in the oyster genome, and found that no homologs of calcite-related shell protein but some homologs of aragonite-related shell proteins in the oyster genome. We explained the results as the new evidence to support that aragonite shells are more ancient than calcite shells in bivalves combined the published biogeological and seawater chemistry data.     

The combined effect of isolation and Fasciola hepatica infection on the life history traits of Fossaria cubensis.

Life history traits of Fossaria cubensis were compared between isolated and paired snails after infection with three miracidia of Fasciola hepatica. Four experimental groups were tested: isolated-unexposed, paired-unexposed, isolated-infected, and paired-infected. A repeated-measures ANOVA showed statistically significant interactions among isolation, infection, and age effects for shell size, number of egg masses per snail, number of eggs per snail, and number of viable eggs per snail. Isolated-unexposed snails exhibited the higher values of these variables and those of survival and finite and intrinsic rates of natural increase. Infection stimulated shell growth during the prepatent period, but differences were present only in paired snails since isolation causes a similar effect. Reproduction, in terms of the number of egg masses per snail and the number of eggs per mass per snail, decreases in the presence of parasitic infection, whereas isolation stimulates it. These effects were observed from early stages of infection.     

Density-dependent growth and reproduction of the apple snail, Pomacea canaliculata: a density manipulation experiment in a paddy field

To examine density dependence in the survival, growth, and reproduction of Pomacea canaliculata, we conducted an experiment in which snail densities were manipulated in a paddy field. We released paint-marked snails of 15–20 mm shell height into 12 enclosures (pens) of 16 m² at one of five densities – 8, 16, 32, 64, or 128 snails per pen. The survival rate of released snails was 95% and was independent of snail density. The snail density had a significant effect on the growth and egg production of individual snails. This density dependence may have been caused by reduced food availability. The females at high density deposited fewer and smaller egg masses than those at low density, and consequently produced fewer eggs. The females at densities 8 and 16 deposited more than 3000 eggs per female, while the females at density 128 oviposited only 414 eggs. The total egg production per pen was, however, higher at higher snail density. The survival rates of juvenile snails were 21%–37% and were independent of adult density. The juvenile density was positively correlated with the total egg production per pen and hence was higher at higher adult density. However, the density of juveniles larger than 5 mm in shell height, i.e., juveniles that can survive an overwintering period, was not significantly different among density treatments. These results suggest that snail density after the overwintering period is independent of the density in the previous year. Thus, density dependence in growth and reproduction might regulate the population of P. canaliculata in paddies. Received: October 23, 1998 / Accepted: July 16, 1999     

Mineralogy of shells from two freshwater snails Belgrandiella fontinalis and B kuesteri

X-ray powder diffraction (XRD) was used to study the mineral composition of shells of snails Belgrandiella fontinalis and Belgrandiella kuesteri collected from three freshwater springs in northeastern Slovenia. The fractions of aragonite, calcite, dolomite and quartz in particular shells were determined. The analysed shells consisted of two or more distinct inorganic layers. The outer shell layer for both species and all sampling localities contained aragonite. The outer layer of B. fontinalis collected at one locality, also contained a small fraction of calcite ( approximately 1 molar%) besides the dominant aragonite. Calcite was identified in the inner layer(s) of both species (2 to 3 molar%), while quartz was found only in B. kuesteri (5-7 molar%). However, both species sampled at one locality showed the presence of dolomite (approx. 20 molar%) in the inner layer(s). The presence of dolomite in the shells of adult gastropods and even molluscs is unusual. A possible formation mechanism and specific ecological factor that could influence the precipitation of dolomite in the shells of different Belgrandiella species is discussed.     

Patterns of shell penetration by Chorus giganteus juveniles (Gastropoda: Muricidae) on the mussel Semimytilus algosus

Patterns are described for shell penetration by the sublittoral muricid snail Chorus giganteus during predatory attacks on the mussel Semimytilus algosus. Location, form and size of shell penetrations were observed in relation to the size of the predator. The results suggested that positions of the perforations on the mussel shells were related to size of the attacking snail. Smaller snails perforated areas near the shell ligament and in the central zone of the shell, while larger snails more frequently attacked shell borders, principally on the ventral side. These observations may be related to: (a) changes in the process of manipulation of the prey during development of the foot and the shell tooth of the predator, (b) changes in internal structure of the snails related to the shell perforation mechanism, or (c) learned behavior acquired experientially by the snails during early growth. Although in other studies of muricid penetration patterns larger boreholes made in shells of the prey were positively correlated with increasing predator size, this relation did not appear to hold with C. giganteus, as larger specimens often made relatively small shell perforations. Areas of boreholes made in the mussel shells by this snail varied from 0.01 to 1.1 mm(2), and were unusually variable in size and shape, especially when compared with literature results on bores characteristic of other muricid species.     

Ultrastructural features and elemental distribution in eggshell during pre and post hatching periods in the green turtle, Chelonia mydas at Ras Al-Hadd, Oman

Eggshells were randomly collected from turtle nests immediately after oviposition and at the end of incubation to examine the ultrastructural features using scanning JSM-5600LV microscopy. Three layers were recognized; an outer calcareous, a middle multistrata and an inner membrane. The calcareous layer had loose nodular units varying in shape and size without interlocking attachments. In freshly laid eggs, each nodular unit had spicules arranged in folded stacks. The spicules became unfolded during incubation, to form radiating configurations. Elemental composition and mapping of the layers were analyzed using energy dispersive spectroscopy (EDS). The elements were unevenly distributed throughout the eggshell and Ca²⁺ decreased significantly after hatching. X-ray diffraction was used to identify the crystals of the eggshells. It revealed that nodular units of the calcareous were made up of CaCO₃, as aragonite (91%), calcite (6%) and vaterite (3%). The middle layer consisted of organic amorphous material with aragonite (89%) and calcite (11%). The shell membrane consisted of reticular fibers with crystals predominantly of NaCl halite. Thermogravimetry analysis of the calcareous layer indicated a complete evaporation of bonded H₂O at 480 °C and CO₂ at 830 °C. Using the differential thermal analysis (DTA), aragonite was transformed to stable calcite at 425 °C.     

Biology and conservation of the rare New Zealand land snail Paryphanta busbyi watti (Mollusca, Pulmonata)

Abstract. The biology of Paryphanta busbyi watti , an endangered carnivorous land snail, was studied mostly by following large juvenile and adult snails with harmonic radar. The snails are nocturnally active and most (79%) hide during the day under leaf litter or in dense vegetation. Fecal analysis showed that the diet is primarily earthworms, but some cannibalism of smaller snails occurs. Empty shells appear to be an additional source of dietary calcium. Mating occurred most frequently between April and July. Mating snails stayed together for 4–7 days, and each pair reversed their positions at least twice. Four snails were first found mating 151–1240 d after they acquired adult shells, and 7 snails were observed mating a second time after 66–298 d. We found 8 nests and observed 6 snails ovipositing; 5 snails laid eggs in holes they dug and one laid eggs in a crevice between rocks. In 2 instances, oviposition was recorded 52 and 140 d after mating. Snails were estimated to lay on average ∼17.5 eggs per year in 3–5 clutches. Most oviposition was observed in August/September, but some occurred between November and February. Of the snails that died, pigs killed 13.6% and humans inadvertently killed another 13.6%. Other snails died from unknown causes mostly during the drier and warmer months, from November to April. This large land snail survives in the presence of introduced predators, but some life history traits could predispose it to a rapid decline in numbers if new predators arrive.     

A Novel Acidic Matrix Protein,PfN44, Stabilizes Magnesium Calcite to Inhibit the Crystallization of Aragonite

Magnesium is widely used to control calcium carbonate deposition in the shell of pearl oysters. Matrix proteins in the shell are responsible for nucleation and growth of calcium carbonate crystals. However, there is no direct evidence supporting a connection between matrix proteins and magnesium. Here, we identified a novel acidic matrix protein named PfN44 that affected aragonite formation in the shell of the pearl oyster Pinctada fucata. Using immunogold labeling assays, we found PfN44 in both the nacreous and prismatic layers. In shell repair, PfN44 was repressed, whereas other matrix proteins were up-regulated. Disturbing the function of PfN44 by RNAi led to the deposition of porous nacreous tablets with overgrowth of crystals in the nacreous layer. By in vitro circular dichroism spectra and fluorescence quenching, we found that PfN44 bound to both calcium and magnesium with a stronger affinity for magnesium. During in vitro calcium carbonate crystallization and calcification of amorphous calcium carbonate, PfN44 regulated the magnesium content of crystalline carbonate polymorphs and stabilized magnesium calcite to inhibit aragonite deposition. Taken together, our results suggested that by stabilizing magnesium calcite to inhibit aragonite deposition, PfN44 participated in P. fucata shell formation. These observations extend our understanding of the connections between matrix proteins and magnesium.     

Shell shape and land-snail habitat in a Mediterranean and desert fauna

The bimodal distribution of shell shape (height: diameter), that is found in various geographically widely separate and taxonomically distinct land snail faunas of many different regions of the world, occurs also in a Mediterranean fauna and in a desert fauna that is derived from it. The desert fauna is, however, closer to the bisector than the Mediterranean one. High-spired snails are mainly rock-dwellers, and equidimensional to low-spired snails are bush-dwellers or soil-diggers, with a few rock-dwellers; litter-dwellers are small-sized species that may have either high- or low-spired shells. These results are discussed in adaptive terms. Litter is probably the more primitive of these micro-environments. Many of the small, litter-dwelling snails are ovo-viviparous rather than oviparous, perhaps so as to avoid attacks on the eggs by saprophytic fungi. The shift away from the litter environment is accompanied by a trend to abandon the ovo-viviparous strategy, in favour of oviparity, the snail using its foot to dig into the soil and lay eggs. The conchometric differences between bush-, ground- and rock-dwelling snails may perhaps reflect selective pressure to increase the size of the foot; and constraints of a habitat that consists of narrow interspaces between rocky boulders. Snails that habitually dig into the ground during periods of inactivity, and roam over the ground when active, requires a very large foot and, consequently, a very large-mouthed shell to accommodate it; the result is an equidimensional shell, globose or turbiniform in shape. Snails that climb up vertical vegetation would also require a large foot, and consequently a large-mouthed shell to contain it. A fully globose shell would however be disadvantageous, since it might cause undesired torque. Hence, bush-dwellers tend to be flatter than soil-diggers. Snails that habitually live in rock crevices, and on hard substrata, would not require a very large foot; they would need a narrow shell, both to enable easy manoeuvring through crevices and to reduce torque, the result being a small-mouthed, usually high-spired shell. The classification of land snails into bush-, soil- or rock-dwellers closely follows the taxonomic classification. In those species that depart from the habitat that is typical of their taxonomic group towards another habitat, the shell alters its shape accordingly.     

Evolutionary relationships among the Pulmonate land snails and slugs (Pulmonata, Stylommatophora)

We have incorporated an additional 56 species of land snails and slugs in our ribosomal (r) RNA molecular phylogeny. The new taxa include representatives of several important groups. The molecular tree now includes 160 species of stylommatophoran land snails and slugs in 144 genera in 61 families. In the rDNA tree, the Stylommatophora are principally divided into an 'achatinoid' and a 'non-achatinoid' clade. Within these clades, several major land snail groups, including the Orthurethra, Elasmognatha, Limacoidea, and Helicoidea, are supported. Overall, the rDNA molecular phylogeny has remained stable following the incorporation of the additional taxa, with these additions having little impact on the major evolutionary patterns in the tree. Taxonomic coverage of the Orthurethra, Orthalicidae, Camaenidae, and Bradybaenidae is increased significantly. The camaenids and bradybaenids form a complex, and both appear to be paraphyletic. Several families of uncertain affinity, such as the Sagdidae and Thyrophorellidae, are included for the first time. The Sagdidae are shown to belong to the Helicoidea, and the Thyrophorellidae to the Achatinoidea.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 593–610.     

Phenotypic plasticity in reproductive traits: importance in the life history of Helix aspersa (Mollusca: Helicidae) in a recently colonized habitat

Reproductive traits of the land snail Helix aspersa Mtiller were investigated under artificial conditions from two samples, one collected from a population exposed to unpredictable human pressures in its natural environment, i.e. a recently created polders area with intensive agriculture, and the other from a snail farm in which animals were reared under constant conditions denned as 'optimal' for growth and reproduction. Results were compared with data collected from natural populations of the same region (Brittany) and from habitats spanning the environmental heterogeneity of the range of the species. A large part of the variation among populations could be explained by different phenotypic covariances between shell size, clutch size and egg size, but not by the number of clutches per snail. Thus, the higher egg production of snails from the polders was related to (i) a strong correlation between clutch size and shell size, shell size being in the upper limit of the overall range for the region concerned; (ii) an uncommonly low egg weight in comparison with the 'norm' of Helix aspersa , this trait seeming to be involved in a trade-off with clutch size. Second clutches were smaller than the first ones, but their eggs were significantly heavier. This difference may be linked to a size-dependent mortality of juveniles during winter which arises in all populations in which hibernation occurs as an adaptation to low temperatures. In addition to the selective regime usually involved for populations of helicid snails from Western Europe, several unpredictable mortality factors occurred in the polders area: herbicide and pesticide treatments (lethal for young snails), human predation (lethal for adults) and burning (letiial for all snails). Life-history patterns of Helix aspersa are discussed in relation to its ability to successfully colonize a large range of habitats modified by humans, to such an extent that it can become a pest.     

Shell mineralogical trends in epifaunal Mesozoic bivalves and their relationship to seawater chemistry and atmospheric carbon dioxide concentration

The Late Triassic-Early Jurassic change from aragonite- to calcite-facilitating conditions in the oceans, which was caused by a decrease of the Mg²⁺/Ca²⁺ ratio of seawater in combination with an increase of the partial pressure of carbon dioxide, also affected the shell mineralogy of epifaunal bivalves. In the “calcite sea” of the Jurassic and Cretaceous, the most diverse and abundant families of epifaunal bivalves had largely calcitic shells. Some of them, such as the Inoceramidae, acquired this shell mineralogy earlier in Earth's history but did not significantly diversify until the onset of “calcite sea” conditions. Others, however, replaced aragonite by calcite in their shell at the beginning of the Jurassic, as shown for the Ostreidae, Gryphaeidae, Pectinidae, Plicatulidae, and Buchiidae. In these families, replacement of aragonite by calcite took place in the middle and inner layer of the shell and was not associated with changes in morphology and life habit. It is therefore proposed that lower metabolic costs rather than higher resistance against dissolution or advantageous physical properties triggered the calcite expansion in their shells. This explanation fits well the observation that clades of thin-shelled bivalves were less affected by the change of seawater chemistry. Thick-shelled clades, by contrast, may suffer a severe decline in diversity until they adapt their shell mineralogy, as demonstrated by the Hippuritoida: The diversity of the Megalodontoidea, which failed to adapt their shell mineralogy to “calcite sea” conditions, dramatically decreased at the end of the Triassic, whereas their descendents became dominant carbonate producers during the Late Mesozoic after they acquired a calcitic outer shell layer in the Late Jurassic. These examples indicate that changes in the seawater chemistry and in the partial pressure of carbon dioxide are factors that influence the diversity of carbonate-secreting animals, and, as in the case of the decline of the Megalodontoidea, may contribute to mass extinctions.     

The effect of aquatic plant abundance on shell crushing resistance in a freshwater snail

Most of the shell material in snails is composed of calcium carbonate but the organic shell matrix determines the properties of calcium carbonate crystals. It has been shown that the deposition of calcium carbonate is affected by the ingestion of organic compounds. We hypothesize that organic compounds not synthesized by the snails are important for shell strength and must be obtained from the diet. We tested this idea indirectly by evaluating whether the abundance of the organic matter that snails eat is related to the strength of their shells. We measured shell crushing resistance in the snail Mexipyrgus churinceanus and the abundance of the most common aquatic macrophyte, the water lily Nymphaea ampla, in ten bodies of water in the valley of Cuatro Ciénegas, Mexico. We used stable isotopes to test the assumption that these snails feed on water lily organic matter. We also measured other factors that can affect crushing resistance, such as the density of crushing predators, snail density, water pH, and the concentration of calcium and phosphorus in the water. The isotope analysis suggested that snails assimilate water lily organic matter that is metabolized by sediment bacteria. The variable that best explained the variation in crushing resistance found among sites was the local abundance of water lilies. We propose that the local amount of water lily organic matter provides organic compounds important in shell biomineralization, thus determining crushing resistance. Hence, we propose that a third trophic level could be important in the coevolution of snail defensive traits and predatory structures.     

The biology of Placostylus ambagiosus (Pulmonata: Bulimulidae) in New Zealand: Part 1. Behaviour,habitat use,abundance, site fidelity,homing and the dimensions of eggs and snails

The large land snail Placostylus ambagiosus (Pulmonata: Bulimulidae) was investigated during a long-term study in northernmost New Zealand. The snails fed at night on fallen leaves from a variety of broadleaf trees and bushes. During the day, most snails in shrubland rested under plants close to their food sources at densities of up to 53 snails per m² and mean live biomasses of 47–72 g/m². Individual snails stayed close to their food plants for up to 12 years, seldom moving away. Resting behaviour and site fidelity in forest was not investigated in detail, but some adult snails in forest returned to their original sites after being moved up to 60 m into a fenced area built to protect them from feral domestic animals. The ovoid eggs (mean 7.0 mm long×5.6 mm wide; ranges 5.5–7.7 mm long; 5.1–6.7 mm wide) were laid in clutches averaging 43 eggs (range 1–113 eggs) in shallow cavities covered with soil. More than one snail may contribute to a clutch. Adult shell height ranged from 43 mm to 97 mm and varied with habitat quality. Where snail abundance was high adults were small. Implications of these results for conservation management are discussed.     

Silent assassins: predation of native New Zealand trichopteran eggs by non-native freshwater gastropods

Aquatic insects that oviposit on rocks may evolve strategies to mitigate loss of eggs to opportunistic predation by common native grazers, but such strategies may be ineffective against non-native grazers. We tested whether the eggs of common New Zealand caddisfly families (Hydrobiosidae and Hydropsychidae) that oviposit on rocks were more susceptible to predation by native or non-native snails. The native snail Potamopyrgus antipodarum (Grey, 1843) and non-native snails Physa acuta Draparnaud, 1805 and Pseudosuccinea columella (Say, 1817) were presented with caddisfly egg masses and the number of individual eggs consumed was recorded after 24 and 48 hours. The larger Pseudosuccinea readily consumed the eggs, especially spumaline-encapsulated Hydrobiosidae eggs. Physa consumed very few Hydrobiosidae eggs and no Hydropsychidae eggs, whereas Potamopyrgus did not consume eggs of either family. The trichopteran egg masses tested did not succumb to predation by the native Potamopyrgus but are highly vulnerable to predation by a larger non-native snail.     

Repeated Mating and Female Fecundity in the Simultaneously Hermaphroditic Land Snail Arianta arbustorum

Summary

To evaluate the influence of repeated mating on female fecundity in the simultaneously hermaphroditic, self-incompatible land snail Arianta arbus-torum, the number and size of clutches, egg size and hatching success of individuals from 3 populations were determined under conditions of isolation and grouping during one breeding season in a field cage experiment. Only adult snails which had mated and oviposited in the preceding year were used.

Sperm storage enabled isolated individuals to continue with the production of fertilized eggs. Snails kept singly or in groups differed neither in number of clutches laid nor in egg size. But isolated snails laid smaller clutches than did grouped snails. As a result isolated snails produced fewer eggs per breeding season. Hatching success varied greatly between populations and rearing conditions. In general, isolated snails had fewer hatchlings than grouped snails, indicating that individuals prevented from remating suffered a reduced fitness.     

厚壳贻贝一种新型贝壳胶原蛋白的重组表达与功能分析

贝壳是一种具有优异力学性能的生物硬组织,贝壳基质蛋白质对贝壳的形成具有重要意义。厚壳贻贝(Mytilus coruscus)贝壳中发现一种类似胶原蛋白质的新型贝壳基质蛋白质,命名为collagen-like protein 2（CLP-2）。然而,该蛋白质的结构与功能以及对贝壳形成的影响机制尚不清楚。为此,本研究对CLP 2开展了序列分析;进一步采取密码子优化结合原核重组表达策略,开展了CLP-2的重组表达;在此基础上分析了重组CLP-2对酸钙结晶的诱导、结晶速率抑制以及碳酸钙结合能力。对CLP-2的序列分析结果表明,该蛋白质序列中含有信号肽及两个Von Willebrand factor A（VWA）结构域。CLP-2在数据库中尚无高同源性蛋白质存在,表明这是一种较为新颖的贝壳基质蛋白。所获得的重组CLP-2对碳酸钙体外结晶表现出明显的诱导作用,扫描电镜以及傅里叶红外光谱结果表明,重组CLP-2可诱导碳酸钙晶体的形貌由立方体形转化为球形,并在高浓度下进一步转化为哑铃形;同时,重组CLP-2可促使碳酸钙晶体的晶型由方解石型向文石型转化;重组CLP-2在体外具有碳酸钙晶体结合作用;此外,重组CLP-2能显著抑制碳酸钙晶体的结晶速度（P<0.01）,并具有浓度依赖性。上述结果表明,厚壳贻贝贝壳CLP-2蛋白质在贝壳,特别是文石型肌棱柱层的生物矿化过程中具有重要作用。上述研究为深入了解贻贝贝壳的形成机制,以及胶原类蛋白质对生物矿化过程的影响奠定了基础。