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Color polymorphism is frequently observed in molluscan shellfish, while the molecular regulation of shell pigmentation is not well understood. Peroxidase is a key enzyme involved in melanogenesis. Here, we identified a heme-peroxidase 2 gene (CgHPX2), and characterized the expression patterns and transcriptional regulation of CgHPX2 in the Pacific oyster Crassostrea gigas. Tissues expression analysis showed that CgHPX2 was a mantle-specific gene and primarily expressed in the edge mantle in black shell color oyster compared with white shell oyster. In situ hybridization showed that strong signals for CgHPX2 were detected in the both inner and outer surface of the outer fold of mantle in the black shell color oyster, whereas positive signals in white shell oyster were mainly localized in the outer surface of the outer fold of mantle. In the embryos and larvae, a high expression level of CgHPX2 was detected in the trochophore stage in both black and white shell color oysters. The temporal localization of CgHPX2 was mainly detected in the shell gland and edge mantle of trochophore and calcified shell larvae, respectively. In addition, a 2227 bp of 5' flanking region sequence of CgHPX2 was cloned, which contained a presumed core promoter region and many potential transcription factor binding sites. Further luciferase assay experiment confirmed that POU domain, class 2, transcription factor 1 (POU2F1), and SRY-box transcription factor 5 (SOX5) were involved in transcriptional regulation of CgHPX2 gene through binding to its specific promoter region. After CgPOU2F1 and CgSOX5 RNA interference, the CgHPX2 gene expression was significantly decreased. These results suggested that CgPOU2F1 and CgSOX5 might be two important transcription factors that positively regulated the expression of CgHPX2 gene, improving our understanding of the transcriptional regulation of molluscan shell pigmentation.

The first step for animals to interact with external environment is to sense. Unlike vertebrate animals with flexibility, it is challenging for ancient animals that are less flexible especially for mollusca with heavy shells. Chiton, as an example, has eight overlapping shells covering almost the whole body, is known to incorporate sensory units called aesthetes inside the shell. We used micro-computed tomography combined with quantitative image analysis to reveal the optimized shell geometry to resist force and the aesthetes' global distribution at the whole animal levels to facilitate sense from diverse directions both in the seawater and air. Additionally, shell proteomics combined with transcriptome reveals shell matrix proteins responsible for shell construction and potentially sensory function, highlighting unique cadherin-related proteins among mollusca. Together, this multi-level evidence of sensory units in the chiton shell may shed light on the formation of chiton shells and inspire the design of hard armor with sensory function.

As biological ceramic composites, mollusk shells exhibit an excellent strength-toughness combination that should be dependent on aragonite/organic matrix interfaces. The mechanical properties and fracture mechanisms of the nacreous structure in the

The Zoological Survey of India, Western Regional Centre, Pune (ZSI-WRC) contains 1887 specimens of mostly unstudied marine brachyuran crabs. The present systematic study on the marine crabs of ZSI-WRC reports 62 species in 44 genera, 18 families and nine superfamilies. Of these, four genera and 13 species are new distributional records within India. We also record three species for the first time from the Indian waters: Metopograpsus cannicci Innocenti, Schubart & Fratini, 2020 (Grapsidae MacLeay, 1838), Varuna yui Hwang & Takeda, 1986 (Varunidae H. Milne Edwards, 1853), and Ilyograpsus paludicola (Rathbun, 1909) (Macrophthalmidae Dana, 1851). The record of Varuna yui is significant as it is the first time it has been found from the western Indian Ocean. The subfamily Ilyograpsinae Števčić, 2005, has likewise not previously been reported from India. The taxonomy of the poorly known menippid crab, Sphaerozius scaber (Fabricius, 1798), is clarified with the designation of a lectotype, and its taxonomy is discussed. Sphaerozius scaber can be distinguished from the only congener S. nitidus Stimpson, 1858, by its prominently tuberculated carapace, palm and carpus of the chelipeds, and the strongly curved male first gonopod, with the distal part gently tapering into a truncate tip. Sphaerozius scaber is a new addition to the west coast of India. With the inclusion of the present new records, India is now known to have 939 species of marine brachyuran crabs in 375 genera of 63 families.

Molluscan bivalves secrete shell matrices into the extrapallial space (EPS) to guide the precipitation of rigid shells. Meanwhile, immune components are present in the EPS and shell matrices, which are pivotal in resistant to invaded pathogens, thus ensuring the shell formation process. However, the origin of these components remains unclear. In this study, we revealed numerous vesicles were secreted from the outer mantle epithelial cells by using light and electron microscopes. The secreted vesicles were isolated by gradient centrifugation and confirmed by transmission electron microscopy. Proteomics analysis showed that the secreted vesicles were composed of cytoplasmic and immune components, most of which do not have signal peptides, indicating that they were secreted by a non-classical pathway. Moreover, real-time PCR revealed that some immune components were highly expressed in the mantle tissue, compared to the hemocytes. FTIR analysis verified the presence of lipids in the shell matrices, indicating that the vesicles have integrated into the shell layers. Taken together, our results suggested that mantle epithelial cells secreted some important immune components into the EPS via secreted vesicle transportation, thus cooperating with the hemocytes to play a vital role in immunity during shell formation.

The new porcellanid crab Polyonyx deezi n. sp. is described on the basis of two specimens from Okinawa Island, Ryukyu Islands, southwestern Japan. The new species belongs to the P. sinensis group and may be closest to P. socialis Werding & Hiller, 2019 in the comparatively broad proportions of the carpi of the chelipeds and meri of the ambulatory legs. However, P. deezi n. sp. is immediately distinguished from all other congeners by the median branchial margins of the carapace being bluntly angular and produced laterally and the dorsal surfaces of the carapace and chelipeds with distinct protuberances. The occurrence of P. deezi n. sp. from coral reefs may be unusual in species of the P. sinensis group because many of the known species have been recorded from estuaries or coastal embayments. An identification key to the Indo-West Pacific species of the Polyonyx sinensis group is provided.

In the present study, we applied Fourier transform infrared (FTIR) and Fourier transform near infrared (FTNIR) spectroscopy to investigate some specific structural aspects of Patella caerulea, Mytilus edulis, Ostrea edulis, and Calista chione shells sampled in different sites. Moreover, for Ostrea edulis and Calista chione, the present study also included fossil samples. As far as FTIR spectroscopy is concerned, the support of statistical and multivariate methods such as the average spectrum (AV), spectral deconvolution, and two-dimensional correlation analysis (2DCOS) allowed to detect structural differences existing within the same mollusc species as a function of the sites they come. These differences can be reasonably linked to the local environmental conditions, which affect the biomineralization pattern of shell formation and growth. These structural differences are related to the calcite, aragonite, Mg-calcite contents, and interactions, as presently observed for fresh and fossil shells. The application of 2DCOS and deconvolution to FTIR spectra also showed the role of the amorphous calcium carbonate (ACC) in the structural characterization of shells, then suggesting the use of a new parameter, the calcite and aragonite to ACC (CAACC) ratio, as a new measurement for the structural characterization of shells. At last, FTNIR spectroscopy allowed detecting the presence of α-helix and β-sheet protein structures in the shells. The results of this study show that also FTIR and FTNIR spectroscopy are able to discern differences in structural characteristics of mollusc shells, a field of environmental studies where scanning electron microscopy and X-ray diffraction are the more widely used methods.

Background Research on mosquito-microbe interactions may lead to new tools for mosquito and mosquito-borne disease control. To date, such research has largely utilized laboratory-reared mosquitoes that typically lack the microbial diversity of wild populations. A logical progression in this area involves working under controlled settings using field-collected mosquitoes or, in most cases, their progeny. Thus, an understanding of how laboratory colonization affects the assemblage of mosquito microbiota would aid in advancing mosquito microbiome studies and their applications beyond laboratory settings. Methods Using high throughput 16S rRNA amplicon sequencing, the internal and cuticle surface microbiota of F1 progeny of wild-caught adult Anopheles albimanus from four locations in Guatemala were characterized. A total of 132 late instar larvae and 135 2–5 day-old, non-blood-fed virgin adult females that were reared under identical laboratory conditions, were pooled (3 individuals/pool) and analysed. Results Results showed location-associated heterogeneity in both F1 larval internal (p = 0.001; pseudo-F = 9.53) and cuticle surface (p = 0.001; pseudo-F = 8.51) microbiota, and only F1 adult cuticle surface (p = 0.001; pseudo-F = 4.5) microbiota, with a more homogenous adult internal microbiota (p = 0.12; pseudo-F = 1.6) across collection sites. Overall, ASVs assigned to Leucobacter, Thorsellia, Chryseobacterium and uncharacterized Enterobacteriaceae, dominated F1 larval internal microbiota, while Acidovorax, Paucibacter, and uncharacterized Comamonadaceae, dominated the larval cuticle surface. F1 adults comprised a less diverse microbiota compared to larvae, with ASVs assigned to the genus Asaia dominating both internal and cuticle surface microbiota, and constituting at least 70% of taxa in each microbial niche. Conclusions These results suggest that location-specific heterogeneity in filed mosquito microbiota can be transferred to F1 progeny under normal laboratory conditions, but this may not last beyond the F1 larval stage without adjustments to maintain field-derived microbiota. These findings provide the first comprehensive characterization of laboratory-colonized F1An. albimanus progeny from field-derived mothers. This provides a background for studying how parentage and environmental conditions differentially or concomitantly affect mosquito microbiome composition, and how this can be exploited in advancing mosquito microbiome studies and their applications beyond laboratory settings.

Shell formation in molluscan bivalves is regulated by organic matrices composed of biological macromolecules, but how these macromolecules assemble in vitro remains elusive. Prismatic layer in the pearl oyster Pinctada fucata consists of polygonal prisms enveloped by thick organic matrices. In this study, we found that the organic matrices were heterogeneously distributed, with highly acidic fractions (EDTA-soluble and EDTA-insoluble) embedded inside the prism columns, while basic EDTA-insoluble faction as inter-column framework enveloping the prisms. The intra-column matrix was enriched in aspartic acid whereas the inter-column matrix was enriched in glycine, tyrosine and phenylalanine. Moreover, the intra-column matrix contained sulfo group further contributing to its acidic property. Proteomics data showed that the intra-column proteins mainly consisted of acidic proteins, while some typical matrix proteins were absent. The absent matrix proteins such as shematrin family and KRMP family were highly basic and contained aromatic amino acids, suggesting that electric charge and hydrophobic effect might play a role in the matrix heterogeneity. Interestingly, chitin metabolism related proteins were abundant in the inter-column matrix, which may be involved in reconstructing the prism organic matrix. Overall, our study suggests that each single prism grew in an enclosed organic envelope and the organic matrix undergoes rearrangement, thus leading to the peculiar growth of the prismatic layer.

We report here hourly variations of Mg/Ca, Sr/Ca, and Ba/Ca ratios in a Mediterranean mussel shell (Mytilus galloprovincialis) collected at the Otsuchi bay, on the Pacific coast of northeastern Japan. This bivalve was living in the intertidal zone, where such organisms are known to form a daily or bidaily growth line comprised of abundant organic matter. Mg/Ca ratios of the inner surface of the outer shell layer, corresponding to the most recent date, show cyclic changes at 25–90 μm intervals, while no interpretable variations are observed in Sr/Ca and Ba/Ca ratios. High Mg/Ca ratios were probably established by (1) cessation of the external supply of Ca and organic layer forming when the shell is closed at low tide, and (2) the strong binding of Mg to the organic layer, but not of Sr and Ba. Immediately following the great tsunami induced by the 2011 Tohoku earthquake, Mg/Ca enrichment occurred, up to 10 times that of normal low tide, while apparent Ba/Ca enrichment was observed for only a few days following the event, therefore serving a proxy of the past tsunami. Following the tsunami, periodic peaks and troughs in Mg/Ca continued, perhaps due to a biological memory effect as an endogenous clock.

The Bythograeidae is unique amongst brachyuran crab taxa as it is the only family where all 6 genera and 16 species are endemic to hydrothermal vents. During the research conducted by German Federal Institute for Geosciences and Natural Resources to identify inactive polymetallic sulphide deposits along Central and Southeast Indian Ridges, the INDEX project collected from hydrothermal vent fields 6 Bythograeidae megalopae. Entire specimens and dissected appendages were stained, mounted on slides and examined using Light Microscopy and Confocal Laser Scanning Microscopy. Additional molecular analysis using mtCOI confirmed the identification of the specimens as Austinograea rodriguezensis Tsuchida & Hashimoto, 2002. The megalopal stage of A. rodriguezensis shows similarities and distinctions between the characters of two other bythograeid megalopae, Bythograea thermydron Williams, 1980, and B. microps Saint Laurent, 1984. Unlike other brachyuran megalopae, B. thermydron and A. rodriguezensis lack long serrate setae and stout serrate spines on the dactylus of the fifth pereiopod. In both species no coxal spines on the pereiopods were observed. The elliptical carapace of B. thermydron is broader than long vs longitudinally rectangular in A. rodriguezensis. This carapace shape resembles B. microps more than B. thermydron, however, the dorsal surface of B. microps carapace is densely covered in short setae vs only covered with short setae on the anterior 1/4 and the posterior 1/6 length of carapace in A. rodriguezensis. Furthermore, the amount of plumose natatory setae on the pleopods in B. microps is in total larger and more variable, than in A. rodriguezensis. Bythograeid megalopae seem quite generalized and miss specific features that reveal them distinctively as endemic vent crab. A distinction from other species is possible by observing the unique combinations of certain characters. This is the first megalopal stage description of Austinograea and the fourth within the Bythograeidae.

The widely recognized color polymorphisms of molluscan shell have been appreciated for hundreds of years by collectors and scientists, while molecular mechanisms underlying shell pigmentation are still poorly understood. Tyrosinase is a key rate-limiting enzyme for the biosynthesis of melanin. Here, we performed an extensive multi-omics data mining and identified two tyrosinase genes, including tyrosinase and tyrosinase-like protein 2 (Tyr and Typ-2 respectively), in the Pacific oyster Crassostrea gigas, and investigated the expression patterns of tyrosinase during adults and embryogenesis in black and white shell color C. gigas. Tissue expression analysis showed that two tyrosinase genes were both specifically expressed in the mantle, and the expression levels of Tyr and Typ-2 in the edge mantle were significantly higher than that in the central mantle. Besides, Tyr and Typ-2 genes were black shell-specific compared with white shell oysters. In situ hybridization showed that strong signals for Tyr were detected in the inner surface of the outer fold, whereas positive signals for Typ-2 were mainly localized in the outer surface of the outer fold. In the embryos and larvae, the high expression of Tyr mRNA was detected in eyed-larvae, while Typ-2 mRNA was mainly expressed at the trochophore and early D-veliger. Furthermore, the tyrosinase activity in the edge mantle was significantly higher than that in the central mantle. These findings indicated that Tyr gene may be involved in shell pigmentation, and Typ-2 is more likely to play critical roles not only in the formation of shell prismatic layer but also in shell pigmentation. In particular, Typ-2 gene was likely to involve in the initial non-calcified shell of trochophores. The work provides valuable information for the molecular mechanism study of shell formation and pigmentation in C. gigas.

Shell color shows broad variation within mollusc species and despite information on the genetic pathways involved in shell construction and color has recently increased, more studies are needed to understand its genetic architecture. The common cockle ( Cerastoderma edule) is a valuable species from ecological and commercial perspectives which shows important variation in shell color across Northeast Atlantic. In this study, we constructed a high-density genetic map, as a tool for screening common cockle genome, which was applied to ascertain the genetic basis of color variation in the species. The consensus genetic map comprised 19 linkage groups (LGs) in accordance with the cockle karyotype (2n = 38) and spanned 1073 cM, including 730 markers per LG and an inter-marker distance of 0.13 cM. Five full-sib families showing segregation for several color-associated traits were used for a genome-wide association study and a major QTL on chromosome 13 associated to different color-traits was detected. Mining on this genomic region revealed several candidate genes related to shell construction and color. A genomic region previously reported associated with divergent selection in cockle distribution overlapped with this QTL suggesting its putative role on adaptation Interreg Atlantic Area | Ref. EAPA_458/2016 Ministerio de Economía, Industria y Competitividad | Ref. BES2016/078166 Roslin Institute | Ref. BBS/E/D/20002172 Roslin Institute | Ref. BBS/E/D/30002275 Roslin Institute | Ref. BBS/E/D/10002070

Assessing the taxonomic importance of the suture line in shelled cephalopods is a key to better understanding the diversity of this group in Earth history. Because fossils are subject to taphonomic artifacts, an in-depth knowledge of well-preserved modern organisms is needed as an important reference. Here, we examine the suture line morphology of all known species of the modern cephalopods Nautilus and Allonautilus. We applied computed tomography and geometric morphometrics to quantify the suture line morphology as well as the conch geometry and septal spacing. Results reveal that the suture line and conch geometry are useful in distinguishing species, while septal spacing is less useful. We also constructed cluster trees to illustrate the similarity among species. The tree based on conch geometry in middle ontogeny is nearly congruent with those previously reconstructed based on molecular data. In addition, different geographical populations of the same species of Nautilus separate out in this tree. This suggests that genetically distinct (i.e., geographically isolated) populations of Nautilus can also be distinguished using conch geometry. Our results are applicable to closely related fossil cephalopods (nautilids), but may not apply to more distantly related forms (ammonoids).

The genus Phylladiorhynchus Baba, 1969 currently contains 11 species, all occurring in the shallow waters and on the continental shelf of the Indian and Pacific oceans. Recent expeditions in these oceans have resulted in the collection of numerous new specimens in need of analysis. We have studied this material using an integrative approach analysing both morphological and molecular (COI and 16S) characters. We describe 41 new species and resurrect three old names: P. integrus (Benedict, 1902) and P. lenzi (Rathbun, 1907), previously synonymized with P. pusillus (Henderson, 1885), and P. serrirostris (Melin, 1939), previously synonymized with P. integrirostris (Dana, 1852). Most species of the genus are described and illustrated. Some species are barely discernible on the basis of morphological characters but are highly divergent genetically. Species of Phylladiorhynchus are mainly distinguishable by the number of epigastric spines and lateral spines of the carapace, the shape and the armature of the rostrum, the number and pattern of the ridges on the carapace and pleon, the shape of thoracic sternite 3 and the armature of the P2–4 dactyli. A dichotomous identification key to all species is provided.

The fossil history of turtle and whale barnacles (Coronuloidea: Chelonibiidae, Platylepadidae, Coronulidae and †Emersoniidae) is fragmentary and has only been investigated in part. Morphological inferences and molecular phylogenetic analyses on extant specimens suggest that the roots of whale barnacles (Coronulidae) are to be found among the chelonibiid turtle barnacles, but the hard-part modifications that enabled early coronuloids to attach to the cetacean skin are still largely to be perceived. Here, we reappraise a fossil chelonibiid specimen from the Miocene of insular Tanzania that was previously referred to the living species Chelonibia caretta. This largely forgotten specimen is here described as the holotype of the new species †Chelonibia zanzibarensis. While similar to C. caretta, †C. zanzibarensis exhibits obvious external longitudinal parietal canals occurring in-between external longitudinal parietal septa that abut outwards to form T-shaped flanges, a character so far regarded as proper of the seemingly more derived Coronulidae and Platylepadidae. Along with these features, the presence of a substrate imprint on the shell exterior indicates that †C. zanzibarensis grasped its host's integument in much the same way as coronulids and platylepadids, albeit without the development of macroscopic parietal buttresses and bolsters. Thin section analyses of the inner parietal architecture of some extant and extinct coronuloids conclusively demonstrate that vestiges of comparable external parietal microstructures are present in some living members of Chelonibiidae. This observation strengthens the unity of Coronuloidea while significantly contributing to our understanding of the evolution of the coronuloid shell structure in adapting to a diverse spectrum of hosts.

Molluscan shell color polymorphism is important in genetic breeding, while the molecular information mechanism for shell coloring is unclear. Here, high-throughput RNA sequencing was used to compare expression profiles of coding and non-coding RNAs (ncRNAs) from Pacific oyster Crassostrea gigas with orange and black shell, which were from an F2 family constructed by crossing an orange shell male with a black shell female. First, 458, 13, and 8 differentially expressed genes (DEGs), lncRNAs (DELs), and miRNAs (DEMs) were identified, respectively. Functional analysis suggested that the DEGs were significantly enriched in 9 pathways including tyrosine metabolism and oxidative phosphorylation pathways. Several genes related to melanin synthesis and biomineralization expressed higher whereas genes associated with carotenoid pigmentation or metabolism expressed lower in orange shell oyster. Then, based on the ncRNA analysis, 163 and 20 genes were targeted by 13 and 8 differentially expressed lncRNAs (DELs) and miRNAs (DEMs), severally. Potential DELs-DEMs-DEGs interactions were also examined. Seven DEMs-DEGs pairs were detected, in which tyrosinase-like protein 1 was targeted by lgi-miR-133-3p and lgi-miR-252a and cytochrome P450 was targeted by dme-miRNA-1-3p. These results revealed that melanin synthesis-related genes and miRNAs-mRNA interactions functioned on orange shell coloration, which shed light on the molecular regulation of shell coloration in marine shellfish.

Biomineralization is a widespread biological process, involved in the formation of shells, teeth, and bones. Shell matrix proteins have been widely studied for their importance during shell formation. In 2015, our group identified 72 unique shell matrix proteins inPinctada fucata, among which PU14 is a matrix protein detected in the soluble fraction that solely exists in the prismatic layer. However, the function of PU14 is still unclear. In this study, the full-length cDNA sequence of PU14 was obtained and functional analyses of PU14 protein during shell formation were performed. The deduced protein has a molecular mass of 77.8 kDa and an isoelectric point of 11.34. The primary protein structure contains Gln-rich and random repeat units, which are typical characteristics of matrix protein and indicate its potential function during shell formation. In vivo and in vitro experiments indicated PU14 has prismatic layer functions during shell formation. The tissue expression patterns showed thatPU14was mainly expressed in the mantle tissue, which is consistent with prismatic layer formation. Notching experiments suggested thatPU14responded to repair and regenerate the injured shell. After inhibiting gene expression by injecting PU14-specific double-stranded RNA, the inner surface of the prismatic layer changed significantly and became rougher. Further, in vitro experiments showed that recombinant protein rPU14 impacted calcite crystal morphology. Taken together, characterization and functional analyses of a novel matrix protein, PU14, provide new insights about basic matrix proteins and their functions during shell formation.

This paper presents the compressive behavior of tortoise carapace at high strain rates and its protection mechanisms under impact loading. Both experimental and numerical results are reported. Tortoise is a land-based desert-dwelling animal taxonomically classified in the order of Testudines. The carapace is the dome-shaped upper part of the tortoise shell that protects its body from predator attacks. The carapace structure is composed of four layers formed as a composite structure with a porous core. The outer surface is keratin scutes made of fibrous structural proteins. The remaining layers are bone-like materials which are dorsal cortex, cancellous interior and ventral cortex. The compressive behavior at high rate of deformation is examined using split Hopkinson pressure bar (SHPB) technique. The results shown in the stress-strain plot illustrate a strain-rate hardening effect. The impact test is conducted using a gas gun with 6.35-mm diameter steel bearing balls as projectiles. The responses of carapace sample under a range of impact velocities are investigated to analyze its protection mechanisms. The numerical model of impact test is created to obtain an insight into mechanical behaviors of the carapace structure that cannot be observed in the experiments. The strain rate dependent material model is defined based on the SHPB test results. The distributions of stress and rebound velocity are presented and discussed.

Background A major goal of evolutionary developmental biology is to discover general models and mechanisms that create the phenotypes of organisms. However, universal models of such fundamental growth and form are rare, presumably due to the limited number of physical laws and biological processes that influence growth. One such model is the logarithmic spiral, which has been purported to explain the growth of biological structures such as teeth, claws, horns, and beaks. However, the logarithmic spiral only describes the path of the structure through space, and cannot generate these shapes. Results Here we show a new universal model based on a power law between the radius of the structure and its length, which generates a shape called a ‘power cone’. We describe the underlying ‘power cascade’ model that explains the extreme diversity of tooth shapes in vertebrates, including humans, mammoths, sabre-toothed cats, tyrannosaurs and giant megalodon sharks. This model can be used to predict the age of mammals with ever-growing teeth, including elephants and rodents. We view this as the third general model of tooth development, along with the patterning cascade model for cusp number and spacing, and the inhibitory cascade model that predicts relative tooth size. Beyond the dentition, this new model also describes the growth of claws, horns, antlers and beaks of vertebrates, as well as the fangs and shells of invertebrates, and thorns and prickles of plants. Conclusions The power cone is generated when the radial power growth rate is unequal to the length power growth rate. The power cascade model operates independently of the logarithmic spiral and is present throughout diverse biological systems. The power cascade provides a mechanistic basis for the generation of these pointed structures across the tree of life.

Various hydroxyapatite (HA) powders synthesized at different temperatures are deposited on titanium alloy by using an atmospheric plasma spray process. These different HA powders were synthesized from Indian clam seashells through the hydrothermal technique at varying temperatures from 700 to 1000 °C for a 2 h time duration in our previous study. The synthesized HA powders are spray-dried to obtain agglomerated powders suitable for spraying during the coating application. Crystallite size, Ca/P ratio, and crystallinity of agglomerated HA powders and their respective coatings are estimated by standard methods. The microstructure and phases of the feedstock and coating materials are investigated by using a field-emission scanning electron microscope (FESEM) and X-ray diffractometer (XRD), respectively. Further, the HA coatings are characterized in terms of surface roughness, microhardness, porosity, adhesion strength, and wear resistance through the stylus profilometer, Vickers micro-hardness tester, image analysis technique, scratch tester, and ball-on-disc tribometer, respectively. The average surface roughness (R

Lytic polysaccharide monooxygenases (LPMOs) catalyze oxidative cleavage of crystalline polysaccharides such as cellulose and chitin and are important for biomass conversion in the biosphere as well as in biorefineries. The target polysaccharides of LPMOs naturally occur in copolymeric structures such as plant cell walls and insect cuticles that are rich in phenolic compounds, which contribute rigidity and stiffness to these materials. Since these phenolics may be photoactive and since LPMO action depends on reducing equivalents, we hypothesized that LPMOs may enable light-driven biomass conversion. Here, we show that redox compounds naturally present in shed insect exoskeletons enable harvesting of light energy to drive LPMO reactions and thus biomass conversion. The primary underlying mechanism is that irradiation of exoskeletons with visible light leads to the generation of H 2 O 2 , which fuels LPMO peroxygenase reactions. Experiments with a cellulose model substrate show that the impact of light depends on both light and exoskeleton dosage and that light-driven LPMO activity is inhibited by a competing H 2 O 2 -consuming enzyme. Degradation experiments with the chitin-rich exoskeletons themselves show that solubilization of chitin by a chitin-active LPMO is promoted by light. The fact that LPMO reactions, and likely reactions catalyzed by other biomass-converting redox enzymes, are fueled by light-driven abiotic reactions in nature provides an enzyme-based explanation for the known impact of visible light on biomass conversion.

The global decline of natural oyster populations emphasizes the need to improve our understanding of their biology. Understanding the role of chemical cues from conspecifics on how oysters occupy appropriate substrata is crucial to learning about their evolution, population dynamics, and chemical communication. Here, a novel role of a macromolecular assembly of shell matrix proteins which act as

Giant clams produce massive calcified shells with important biological (e.g., defensive) and ecological (e.g., habitat-forming) properties. Whereas elevated seawater temperature is known to alter giant clam shell structure, no study has examined the effects of a simultaneous increase in seawater temperature and partial pressure of carbon dioxide (pCO2) on shell mineralogical composition in these species. We investigated the effects of 60-days exposure to end-of-the-century projections for seawater temperature (+ 3 °C) and pCO2 (+ 500 µatm) on growth, mineralogy, and organic content of shells and scutes in juvenile Tridacna squamosa giant clams. Elevated temperature had no effect on growth rates or organic content, but did increase shell [24Mg]/[40Ca] as well as [40Ca] in newly-formed scutes. Elevated pCO2 increased shell growth and whole animal mass gain. In addition, we report the first evidence of an effect of elevated pCO2 on element/Ca ratios in giant clam shells, with significantly increased [137Ba]/[40Ca] in newly-formed shells. Simultaneous exposure to both drivers greatly increased inter-individual variation in mineral concentrations and resulted in reduced shell N-content which may signal the onset of physiological stress. Overall, our results indicate a greater influence of pCO2 on shell mineralogy in giant clams than previously recognized.

Turtles are characterized by their typical carapace, which is primarily composed of corneous beta proteins in the horny part and collagen in the dermal part. The formation of the extracellular matrix in the dermis of the carapace in a hard-shelled and a soft-shelled turtle has been compared. The study examines carapace development, with an emphasis on collagen accumulation, in the soft-shelled turtle Pelodiscus sinensis and hard-shelled turtle Trachemys scripta elegans, using comparative morphological and embryological analyses. The histological results showed that collagen deposition in the turtle carapace increased as the embryos developed. However, significant differences were observed between the two turtle species at the developmental stages examined. The microstructure of the dermis of the carapace of P. sinensis showed light and dark banding of collagen bundles, with a higher overall collagen content, whereas the carapacial matrix of T. scripta was characterized by loosely packed and thinner collagenous fiber bundles with a lower percentage of type I collagen. Overall, the formation and distribution of collagen fibrils at specific developmental stages are different between the soft-and hard-shelled turtles. These results indicate that the pliable epidermis of the soft-shelled turtle is supported by a strong dermis that is regularly distributed with collagen and that it allows improved maneuvering, whereas a strong but inflexible epidermis as observed in case of hard-shelled turtles limits movement. This article is protected by copyright. All rights reserved.

Precisely targeted measurements of trace elements using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) reveal inter-chamber heterogeneities in specimens of the planktic foraminifer Trilobatus (Globigerinoides) sacculifer. We find that Mg/Ca ratios in the final growth chamber are generally lower compared to previous growth chambers, but final chamber Mg/Ca is elevated in one of thirteen sample intervals. Differences in distributions of Mg/Ca values from separate growth chambers are observed, occurring most often at lower Mg/Ca values, suggesting that single-chamber measurements may not be reflective of the specimen’s integrated Mg/Ca. We compared LA-ICPMS Mg/Ca values to paired, same-individual Mg/Ca measured via inductively coupled plasma optical emission spectrometry (ICP-OES) to assess their correspondence. Paired LA-ICPMS and ICP-OES Mg/Ca show a maximum correlation coefficient of R = 0.92 (p

Body elongation is a general feature of development. Postembryonically, the body needs to be framed and protected by extracellular materials, such as the skeleton, the skin and the shell, which have greater strength than cells. Thus, body elongation after embryogenesis must be reconciled with those rigid extracellular materials. Here we show that the exoskeleton (cuticle) coating the Drosophila larval body has a mechanical property to expand less efficiently along the body circumference than along the anteroposterior axis. This “corset” property of the cuticle directs a change in body shape during body growth from a relatively round shape to an elongated one. Furthermore, the corset property depends on the functions of Cuticular protein 11 A and Tubby, protein components of a sub-surface layer of the larval cuticle. Thus, constructing a stretchable cuticle and supplying it with components that confer circumferential stiffness is the fly’s strategy for executing postembryonic body elongation., Tajiri et al. describe how the cuticle coating the Drosophila larval body expands less efficiently along the body circumference than along the anteroposterior axis to drive body elongation. This “corset” property depends on cuticular proteins Cpr11A and Tubby, which may work together to change larval body shape.

The C1q protein, which contains the globular C1q (gC1q) domain, is involved in the innate immune response, and is found abundantly in the shell, and it participates in the shell formation. In this study, a novel gC1q domain-containing gene was identified from Pinctada fucata martensii (P. f. martensii) and designated as PmC1qDC-1. The full-length sequence of PmC1qDC-1 was 902 bp with a 534 bp open reading frame (ORF), encoding a polypeptide of 177 amino acids. Quantitative real-time PCR (qRT-PCR) result showed that PmC1qDC-1 was widely expressed in all tested tissues, including shell formation-associated tissue and immune-related tissue. PmC1qDC-1 expression was significantly high in the blastula and gastrula and especially among the juvenile stage, which is the most important stage of dissoconch shell formation. PmC1qDC-1 expression was located in the outer epithelial cells of mantle pallial and mantle edge and irregular crystal tablets were observed in the nacre upon knockdown of PmC1qDC-1 expression at mantle pallial. Moreover, the recombined protein PmC1qDC-1 increased the rate of calcium carbonate precipitation. Besides, PmC1qDC-1 expression was significantly up-regulated in the mantle pallial at 6 h and was significantly up-regulated in the mantle edge at 12 h and 24 h after shell notching. The expression level of PmC1qDC-1 in mantle edge was significantly up-regulated at 48 h after LPS stimulation and was significantly up-regulated at 12 h, 24 h and 48 h after poly I:C stimulation. Moreover, PmC1qDC-1 expression was significantly up-regulated in hemocytes at 6 h after lipopolysaccharide (LPS) and poly I:C challenge. These findings suggest that PmC1qDC-1 plays a crucial role both in the shell formation and the innate immune response in pearl oysters, providing new clues for understanding the shell formation and defense mechanism in mollusk.

Pteropods, a group of holoplanktonic gastropods, are regarded as bioindicators of the effects of ocean acidification on open ocean ecosystems, because their thin aragonitic shells are susceptible to dissolution. While there have been recent efforts to address their capacity for physiological acclimation, it is also important to gain predictive understanding of their ability to adapt to future ocean conditions. However, little is known about the levels of genetic variation and large‐scale population structuring of pteropods, key characteristics enabling local adaptation. We examined the spatial distribution of genetic diversity in the mitochondrial cytochrome c oxidase I (COI) and nuclear 28S gene fragments, as well as shell shape variation, across a latitudinal transect in the Atlantic Ocean (35°N–36°S) for the pteropod Limacina bulimoides. We observed high levels of genetic variability (COI π = 0.034, 28S π = 0.0021) and strong spatial structuring (COI ΦST = 0.230, 28S ΦST = 0.255) across this transect. Based on the congruence of mitochondrial and nuclear differentiation, as well as differences in shell shape, we identified a primary dispersal barrier in the southern Atlantic subtropical gyre (15–18°S). This barrier is maintained despite the presence of expatriates, a gyral current system, and in the absence of any distinct oceanographic gradients in this region, suggesting that reproductive isolation between these populations must be strong. A secondary dispersal barrier supported only by 28S pairwise ΦST comparisons was identified in the equatorial upwelling region (between 15°N and 4°S), which is concordant with barriers observed in other zooplankton species. Both oceanic dispersal barriers were congruent with regions of low abundance reported for a similar basin‐scale transect that was sampled 2 years later. Our finding supports the hypothesis that low abundance indicates areas of suboptimal habitat that result in barriers to gene flow in widely distributed zooplankton species. Such species may in fact consist of several populations or (sub)species that are adapted to local environmental conditions, limiting their potential for adaptive responses to ocean changes. Future analyses of genome‐wide diversity in pteropods could provide further insight into the strength, formation and maintenance of oceanic dispersal barriers., Shelled pteropods or 'sea butterflies' are proposed as bio‐indicators to monitor impacts of ocean acidification on open ocean ecosystems. Local adaptation is affected by the presence and geographic extent of distinct populations, however, little is known about the large‐scale population structure of pteropods. We thus examined the spatial distribution of genetic and morphometric variation in the species Limacina bulimoides across the Atlantic Ocean. Two dispersal barriers were identified that were congruent with regions of low abundance, suggesting that regions of suboptimal habitat may create barriers to gene flow in widely‐distributed zooplankton species.

Many biological structures use liquid crystals as self-organizing templates for their formation. We review and analyse evidence that the crossed-lamellar biomineral microstructure of mollusc shells may be formed from such a liquid-crystal precursor., We acknowledge funding from projects CGL2017-85118-P and FIS2016-77692-C2-2-P of the Spanish Ministerio de Ciencia e Innovación, the Unidad Científica de Excelencia UCE-PP2016-05 of the University of Granada and the Research Group RNM363 of the Junta de Andalucía.