Structure and mineralization of the spearing mantis shrimp (Stomatopoda; Lysiosquillina maculata) body and spike cuticles.

Stomatopoda is a crustacean order including sophisticated predators called spearing and smashing mantis shrimps that are separated from the well-studied Eumalacotraca since the Devonian. The spearing mantis shrimp has developed a spiky dactyl capable of impaling fishes or crustaceans in a fraction of second. In this high velocity hunting technique, the spikes undergo an intense mechanical constraint to which their exoskeleton (or cuticle) has to be adapted. To better understand the spike cuticle internal architecture and composition, electron microscopy, X-ray microanalysis and Raman spectroscopy were used on the spikes of 7 individuals (collected in French Polynesia and Indonesia), but also on parts of the body cuticle that have less mechanical stress to bear. In the body cuticle, several specificities linked to the group were found, allowing to determine the basic structure from which the spike cuticle has evolved. Results also highlighted that the body cuticle of mantis shrimps could be a model close to the ancestral arthropod cuticle by the aspect of its biological layers (epi- and procuticle including exo- and endocuticle) as well as by the Ca-carbonate/phosphate mineral content of these layers. In contrast, the spike cuticle exhibits a deeply modified organization in four functional regions overprinted on the biological layers. Each of them has specific fibre arrangement or mineral content (fluorapatite, ACP or phosphate-rich Ca-carbonate) and is thought to assume specific mechanical roles, conferring appropriate properties on the entire spike. These results agree with an evolution of smashing mantis shrimps from primitive stabbing/spearing shrimps, and thus also allowed a better understanding of the structural modifications described in previous studies on the dactyl club of smashing mantis shrimps.

Heterocypris spadix sp. nov. (Crustacea: Ostracoda: Cypridoidea) from Japan, with Information on Its Reproductive Mode.

We describe the cypridoidean ostracod Heterocypris spadix sp. nov. from brackish water on Okinawa Island, Japan. The species closely resembles Heterocypris salina (Brady, 1868) but differs in that (1) the marginal infolds on valves are less developed, (2) the tubercles on the anterior margin of the right valve are completely covered by the selvage and invisible in inner view, and (3) the calcified inner lamella on the ventral margin of the left and right valves is scarcely evident in inner view, as the ventral margins of the valves bend inwardly. We determined partial sequences for the cytochrome c oxidase subunit I (COI; cox1) and 18S rRNA genes in H. spadix for future DNA barcoding and phylogenetic analyses. Our sample contained only females. A breeding experiment revealed that H. spadix females reproduce parthenogenetically. Another experiment showed that H. spadix has low tolerance to desiccation, with all individuals at 25°C dying between 1-2 hours after removal from water. We amplified and sequenced a partial 16S rRNA sequence for the endosymbiotic bacterium Cardinium from H. spadix. Infection by Cardinium may be related to the parthenogenetic reproductive mode we observed in H. spadix.

More than one way to smell ashore - Evolution of the olfactory pathway in terrestrial malacostracan crustaceans.

Crustaceans provide a fascinating opportunity for studying adaptations to a terrestrial lifestyle because within this group, the conquest of land has occurred at least ten times convergently. The evolutionary transition from water to land demands various morphological and physiological adaptations of tissues and organs including the sensory and nervous system. In this review, we aim to compare the brain architecture between selected terrestrial and closely related marine representatives of the crustacean taxa Amphipoda, Isopoda, Brachyura, and Anomala with an emphasis on the elements of the olfactory pathway including receptor molecules. Our comparison of neuroanatomical structures between terrestrial members and their close aquatic relatives suggests that during the convergent evolution of terrestrial life-styles, the elements of the olfactory pathway were subject to different morphological transformations. In terrestrial anomalans (Coenobitidae), the elements of the primary olfactory pathway (antennules and olfactory lobes) are in general considerably enlarged whereas they are smaller in terrestrial brachyurans compared to their aquatic relatives. Studies on the repertoire of receptor molecules in Coenobitidae do not point to specific terrestrial adaptations but suggest that perireceptor events - processes in the receptor environment before the stimuli bind - may play an important role for aerial olfaction in this group. In terrestrial members of amphipods (Amphipoda: Talitridae) as well as of isopods (Isopoda: Oniscidea), however, the antennules and olfactory sensilla (aesthetascs) are largely reduced and miniaturized. Consequently, their primary olfactory processing centers are suggested to have been lost during the evolution of a life on land. Nevertheless, in terrestrial Peracarida, the (second) antennae as well as their associated tritocerebral processing structures are presumed to compensate for this loss or rather considerable reduction of the (deutocerebral) primary olfactory pathway. We conclude that after the evolutionary transition from water to land, it is not trivial for arthropods to establish aerial olfaction. If we consider insects as an ingroup of Crustacea, then the Coenobitidae and Insecta may be seen as the most successful crustacean representatives in this respect.

Establishment of long-term ostracod epidermal culture.

Primary crustacean cell culture was introduced in the 1960s, but to date limited cell lines have been established. Skogsbergia lerneri is a myodocopid ostracod, which has a body enclosed within a thin, durable, transparent bivalved carapace, through which the eye can see. The epidermal layer lines the inner surface of the carapace and is responsible for carapace synthesis. The purpose of the present study was to develop an in vitro epidermal tissue and cell culture method for S. lerneri. First, an optimal environment for the viability of this epidermal tissue was ascertained, while maintaining its cell proliferative capacity. Next, a microdissection technique to remove the epidermal layer for explant culture was established and finally, a cell dissociation method for epidermal cell culture was determined. Maintenance of sterility, cell viability and proliferation were key throughout these processes. This novel approach for viable S. lerneri epidermal tissue and cell culture augments our understanding of crustacean cell biology and the complex biosynthesis of the ostracod carapace. In addition, these techniques have great potential in the fields of biomaterial manufacture, the military and fisheries, for example, in vitro toxicity testing.

A new internal structure of nauplius larvae: A "ghostly" support sling for cypris y left within the exuviae of nauplius y after metamorphosis (Crustacea: Thecostraca: Facetotecta).

Facetotecta, or crustacean "y-larvae," occur in all the world's oceans although the adult forms remain completely unknown. At the metamorphic molt from the last naupliar instar to the terminal cypris larval stage a free carapace, six pairs of natatory thoracopods, and a segmented thorax and abdomen all develop anew. Unlike in earlier molts, the cephalic shield and the so-called "faciotruncal integument" usually remain together at this last naupliar molt, and the posterior "trunk" portion of the exuviae, while hollow, is not empty. In mounted preparations examined by phase contrast or differential interference contrast (DIC) microscopy, a ghost-like image of part of the cypris thorax, particularly the thoracopods and even their setae, is commonly visible inside the naupliar exuviae, and may be universally present in the Facetotecta. To investigate this "ghost," we used DIC and digital photographic stacking, and also scanning electron microscopy, on slide or stub-mounted final naupliar exuviae of an assortment of undescribed species of Facetotecta that had been reared from planktonic lecithotropic nauplii to the cypris stage at Sesoko Island, Okinawa, Japan, and at Keelung and Green Island, Taiwan. These techniques showed that the "ghost" is a delicate, three-dimensional, fibrous structure, essentially a sling-like mold or matrix with struts attached to the outer cuticle and pairs of deep pockets that previously held the thoracopods of the developing cypris y. Whether it is endoskeletal in nature, the (partial) exuvia of an additional instar, remnants of apoptosis, or something else is currently unknown. Nothing similar has been reported in other thecostracans, or in other crustaceans that undergo a similarly abrupt metamorphosis at the last naupliar molt.

Ultrastructure of spermiogenesis and the distribution of spermatozoal nuclear histones in the Japanese mantis shrimp, Oratosquilla oratoria (Crustacea: Stomatopoda).

The Japanese mantis shrimp Oratosquilla oratoria (Stomatopoda; Crustacea) is one of the most economically important aquatic species of Pacific shrimp and it is distributed from Japan to the coast of China, the Philippines, the Malay Peninsula, and the Hawaiian Islands. Early studies described certain characteristics of spermatogenesis and the sperm ultrastructure in Stomatopoda, but the composition of sperm basic nuclear proteins (SBNPs) remains completely unknown. We studied the sperm ultrastructure of O. oratoria using transmission electron microscopy and the histone composition using immunofluorescence and immunoelectron microscopy. We found that the spherical nucleus is adjacent to the electron translucent external coat, which occurs in early spermatids. The acrosomal structure begins to form at the junction of the nucleus and the external coat. At the mid-spermatid stage, part of the chromatin appears to be more electron-dense than the external coat side. The aflagellate sperm of O. oratoria, are rounded or slightly ovoid in shape and have a consistent granular nucleus, an acrosome structure of pushpin shape and a spherical vesicular body in which faintly granular material is scattered. The acrosome consists of an acrosomal vesicle, perforatorium, and subacrosomal material. The sperm contains histones H2A, H2B, H3, H4, H3.3, H2AX, and H2AZ as well as some histone modifications, that is, H3K9me3, H3K4me2, H3S10ph, H4Kac, and H2A + H4S1ph. Histones are localized not only in the nucleus of the sperm but also in other structures outside the nucleus. The results may provide new perspectives for systematic studies of crustaceans and their sperm chromatin components. These findings extend the study of the sperm structure of Stomatopoda and provide basic data to elucidate the epigenetic mechanism of fertilization.

Ultrastructure and cuticle formation of the carapace in the myodocopan ostracod exemplified by Euphilomedes japonica (Crustacea: Ostracoda).

The ultrastructure and formation of the cuticle of a myodocopan ostracod, Euphilomedes japonica, are investigated utilizing scanning and transmission electron microscopy. The outer lamella cuticle consists of four layers; epicuticle, exocuticle, endocuticle, and membranous layer like in the cuticle of other arthropods. The exocuticle and endocuticle are well-calcified and the organic matrix develops within the both cuticles. The outermost layer of new cuticle (epicuticle) is secreted first and the inner layers (exocuticle, endocuticle and membranous layer) are added proximally in the pre-, and postmoult stages. The calcification takes place in the whole area of carapace at the same time together with the synthesis of organic matrix within the endocuticle. This study demonstrates that the ultrastructure and formation of the cuticle in myodocopans are different from those in podocopans, and that the myodocopan carapaces have achieved a structural diversity for adaptation to different lifestyles.

Ostracoda and foraminifera from Paleocene (Olinda well), Paraíba Basin, Brazilian Northeast

ABSTRACT Paleocene ostracods and planktonic foraminifera from the Maria Farinha Formation, Paraíba Basin, are herein presented. Eleven ostracod species were identified in the genera Cytherella Jones, Cytherelloidea Alexander, Eocytheropteron Alexander, Semicytherura Wagner, Paracosta Siddiqui, Buntonia Howe, Soudanella Apostolescu, Leguminocythereis Howe and, probably, Pataviella Liebau. The planktonic foraminifera are represented by the genera Guembelitria Cushman, Parvularugoglobigerina Hofker, Woodringina Loeblich and Tappan, Heterohelix Ehrenberg, Zeauvigerina Finlay, Muricohedbergella Huber and Leckie, and Praemurica Olsson, Hemleben, Berggren and Liu. The ostracods and foraminifera analyzed indicate an inner shelf paleoenvironment for the studied section. Blooms of Guembelitria spp., which indicate either shallow environments or upwelling zones, were also recorded reinforcing previous paleoenvironmental interpretations based on other fossil groups for this basin.

Ostracoda and foraminifera from Paleocene (Olinda well), Paraíba Basin, Brazilian Northeast.

Paleocene ostracods and planktonic foraminifera from the Maria Farinha Formation, Paraíba Basin, are herein presented. Eleven ostracod species were identified in the genera Cytherella Jones, Cytherelloidea Alexander, Eocytheropteron Alexander, Semicytherura Wagner, Paracosta Siddiqui, Buntonia Howe, Soudanella Apostolescu, Leguminocythereis Howe and, probably, Pataviella Liebau. The planktonic foraminifera are represented by the genera Guembelitria Cushman, Parvularugoglobigerina Hofker, Woodringina Loeblich and Tappan, Heterohelix Ehrenberg, Zeauvigerina Finlay, Muricohedbergella Huber and Leckie, and Praemurica Olsson, Hemleben, Berggren and Liu. The ostracods and foraminifera analyzed indicate an inner shelf paleoenvironment for the studied section. Blooms of Guembelitria spp., which indicate either shallow environments or upwelling zones, were also recorded reinforcing previous paleoenvironmental interpretations based on other fossil groups for this basin.

Formation and function of the "Xestoleberis-spot" in Xestoleberis hanaii (Crustacea: Ostracoda).

The crescent sculpture of the so-called "Xestoleberis-spot" develops inside the calcified valve of the family Xestoleberididae. Electron microscopic observations on both, intermoult and postmoult stages of Xestoleberis species reveal that the "Xestoleberis-spot" system consists of three elements; two calcified chambers, a vesicle of electron-dense material and an uncalcified procuticle. The formation and function of the "Xestoleberis-spot" system are discussed. In conclusion, the "Xestoleberis-spot" system functions as the muscle attachment site for several antennal muscles, and provides the material for chitinous fibers in the exocuticle of outer lamella. The unique cuticular structures of the family Xestoleberididae are due to the "Xestoleberis-spot" system.

A complete survey of normal pores on a smooth shell ostracod (Crustacea): Landmark-based versus outline geometric morphometrics.

Pores and sensilla on ostracod shell have often been used in studies of ontogeny, taxonomy, and phylogeny of the group. However, an analysis of sexual dimorphism and variation between valves in the number and distribution of pores is lacking. Also, such studies have never been done on a widely distributed, morphologically variable, and weakly ornamented freshwater ostracod. Here, we survey pores in one such species, Physocypria kraepelini. We choose 27 homologous pores as landmarks for 2D-geometric morphometric analysis, with the aim to assess intersexual and between valves variation in size and shape relative to the Fourier outline analysis. This species has only simple (Type A) pores with and without a lip, and each pore carries an undivided sensory seta. Our results show that the total number of pores varies (from 270 to 296), but this is not associated with a specific valve. Males carry fewer pores than females, however no sex specific pores are found. Small intrapopulation divergence of the Cyt b molecular marker (1%) indicates that morphological variability is not species related. We found that P. kraepelini exhibits directional asymmetry of size and shape, sexual size dimorphism (SSD) but lacks sexual shape dimorphism (SShD). Two geometric morphometrics methods were congruent in the estimation of SSD, SShD, and directional asymmetry of shape but differ in the statistical evaluation of directional asymmetry of size. Contrary to other animal groups, our study suggests that ostracods have more pronounced directional asymmetry of shape compared to directional asymmetry of size.

Functional morphology of amplexus (clasping) in spinicaudatan clam shrimps (Crustacea, Branchiopoda) and its evolution in bivalved branchiopods: A video-based analysis.

Male clam shrimps (Crustacea: Branchiopoda: Laevicaudata, Spinicaudata, and Cyclestherida) have their first one or two trunk limb pairs modified as "claspers," which are used to hold the female during mating and mate guarding. Clasper morphology has traditionally been important for clam shrimp taxonomy and classification, but little is known about how the males actually use the claspers during amplexus (clasping). Homologies of the various clasper parts ("movable finger," "large palp," "palm," "gripping area," and "small palp") have long been discussed between the three clam shrimp taxa, and studies have shown that only some structures are homologous while others are convergent ("partial homology"). We studied the clasper functionality in four spinicaudatan species using video recordings and scanning electron microscopy, and compared our results with other clam shrimp groups. General mating behavior and carapace morphology was also studied. Generally, spinicaudatan and laevicaudatan claspers function similarly despite some parts being nonhomologous. We mapped clasper morphology and functionality aspects on a branchiopod phylogeny. We suggest that the claspers of the three groups were adapted from an original, simpler clasper, each for a "stronger" grip on the female's carapace margin: 1) Spinicaudata have two clasper pairs bearing an elongated apical club/gripping area with one setal type; 2); Cyclestherida have one clasper pair with clusters of molariform setae on the gripping area and at the movable finger apex; and 3) Laevicaudata have one clasper pair, but have incorporated an additional limb portion into the clasper palm and bear a diverse set of setae. J. Morphol. 278:523-546, 2017. © 2017 Wiley Periodicals, Inc.

Removal of extracellular coat from giant sperm in female receptacle induces sperm motility in Mytilocypris mytiloides (Cyprididae, Ostracoda, Crustacea).

Previous studies of cypridoidean ostracods have noted that (1) their giant spermatozoa are immotile inside the male, (2) these spermatozoa are motile in the female seminal receptacle and (3) these receptacles are often filled with empty sperm coats. Such findings have led previous authors to hypothesize that sperm must shed their coats in the female receptacle to become motile. We present light and electron microscopy results and video recordings of mating experiments with virgin specimens of Mytilocypris mytiloides. We show that the empty sperm coats frequently found in the female receptacles are not the result of sperm molting but are the resistant inner coats of exhausted sperm not used for egg fertilization. In contrast, we show that an outer granular coating material is successively removed from the sperm while resident inside the female receptacles before first oviposition occurs. During this period, previously immotile sperm gain motility, showing strong movement shortly before first oviposition takes place. By correlation of these phenomena, we suggest that dissolution of the outer coat material is required for motility to develop.

Seven new species of the genus Xestoleberis (Ostracoda: Podocopida: Cytheroidea) from the Fiji Archipelago.

The genus Xestoleberis has a global distribution, and although they are predominant in shallow marine environments adapted to both sediment and algal habitats, only two species of this genus, Xestoleberis curta (Brady, 1866) and Xestoleberis variegata Brady, 1880, have previously been reported from the Fiji archipelago. Herein we report seven new species of the genus Xestoleberis from intertidal environments of fringing reef flats of the Fiji Islands: Xestoleberis becca n. sp., Xestoleberis concava n. sp., Xestoleberis gracilariaii n. sp., Xestoleberis marcula n. sp., Xestoleberis natuvuensis n. sp., Xestoleberis penna n. sp. and Xestoleberis petrosa n. sp. With the exception of X. becca n. sp., Xestoleberis species show restricted distribution within Fijian waters. The possible causes for their distribution patterns are suggested to be physical barriers imposed by the fast flowing Bligh Water currents, and islands separated by deep ocean waters.

Stress physiology and weapon integrity of intertidal mantis shrimp under future ocean conditions.

Calcified marine organisms typically experience increased oxidative stress and changes in mineralization in response to ocean acidification and warming conditions. These effects could hinder the potency of animal weapons, such as the mantis shrimp's raptorial appendage. The mechanical properties of this calcified weapon enable extremely powerful punches to be delivered to prey and aggressors. We examined oxidative stress and exoskeleton structure, mineral content, and mechanical properties of the raptorial appendage and the carapace under long-term ocean acidification and warming conditions. The predatory appendage had significantly higher % Mg under ocean acidification conditions, while oxidative stress levels as well as the % Ca and mechanical properties of the appendage remained unchanged. Thus, mantis shrimp tolerate expanded ranges of pH and temperature without experiencing oxidative stress or functional changes to their weapons. Our findings suggest that these powerful predators will not be hindered under future ocean conditions.

Fine morphology of frontal filaments in nauplii of cirriped crustaceans.

Fine morphology of the frontal filaments (FFs) at all nauplius stages of two barnacle species (Verruca stroemia and Hesperibalanus hesperius) has been investigated by scanning electron microscopy. FFs have been detected at the second nauplius stage and persist during all stages. FFs contain a wide proximal and a fine distal parts, but they are not actually separated as segments of the limbs, and the area between them looks like a single cuticular crease. Apical and subapical pores have been found at the top of each FF in the larvae of both species, which may indicate the chemoreceptor function of these organs.

Testis follicles ultrastructure of three species of terrestrial isopods (Crustacea, Isopoda Oniscidea).

The aim of the research, carried out on three species of terrestrial isopods - Armadillidium granulatum, Halophiloscia hirsuta and Trichoniscus alexandrae - is to bring a first consistent contribution to the knowledge of the ultrastructural organization of the testis follicles. The testis follicles are seat of a remarkable dynamic activity of their cell components (somatic cells and germ cells) that results in a continuous variation, related to the trend of spermatogenesis, of their morphology, organization and of the relationships between the two cell populations. The somatic cells, known in literature as follicular cells, nurse cells or Sertoli cells, are arranged at the periphery of the follicle to form an epithelial layer of variable thickness resting on a thin basal lamina in turn surrounded by a discontinuous network of muscle cells. In A. granulatum and H. hirsuta, two types of Sertoli cells are present: a first type, the nurse cells, envelop the spermatids in cavities within their cytoplasm and through their secretion activity play a fundamental role in the formation of the spermatophores; moreover, they phagocytizes the residual cytoplasm of spermatids. A second type of Sertoli cells shows features that leave clearly identify its supporting role to the spermatophores in formation. In T. alexandrae, instead, only one type of Sertoli cells, the nurse cell, is present, whose features are widely superimposable to those observed in the other two species. Moreover, two septa of Sertoli cells depart from the periphery of the testis follicle to constitute an articulated compartmentalization of the follicle itself, probably targeted to realize at its inside a series of microenvironments functionally diversified in order to meets the needs of the different stages of the spermatogenic cycle.

Gomphocythere besni n. sp. (Crustacea, Ostracoda) from a man-made pool (Adiyaman, Turkey).

We describe a new species (Gomphocythere besni n. sp.) from the Tavas man-made pool in Besni town (Adiyaman, Turkey). The species has several differences from its congeners in furcal structures (forked organ, seta of caudal ramus), numbers of setae on maxillula and maxillular palp, particular ornamentation with up to eight fossae in each mesh of the reticulation and in the presence of two types of lateral pore-canals (single pore opening with a sensillum and sieve plates) on the carapace. The finding of the new species extends the known geographical distribution of the living forms of the genus further to the North. The new species was found from relatively cool (16.6 ºC) and medium oxygenated waters (7.36 mg/L) in a mixture of sand and gravel substrate. Details about its ecology and taxonomic status are also compared and discussed with other species of the same genus.

Five Sarsiellidae ostracods (Crustacea: Myodocopida) from the South Coast of Korea (East China Sea).

The East China Sea is part of the Warm Temperate Northwest Pacific zoogeographic province and, as such, has a high biodiversity and many tropical and subtropical biotic elements. Nevertheless, many invertebrate groups from this area remain poorly studied. Ostracods are one of them, especially those belonging to the subclass Myodocopa. In this paper we provide the first data on a diverse myodocopid family, Sarsiellidae, not only for the East China Sea, but also for Korea. Five species are reported in this paper from three Korean islands (Jeju, Chuja, and Maemul), and they are only a part of the ostracods collected during this study, indicating a high diversity of the group in this region. Three new species, Eurypylus koreanus sp. nov., Eusarsiella hanguk sp. nov., and Sarsiella nereis sp. nov., clearly stand apart from their respective congeners, mostly by prominent shell characters but also by details of the soft part morphology. Their affinity though clearly indicates a close connection of the region with the more southern zoogeographical realms, especially Central Indo Pacific and partly Temperate Australasia. Two species previously known from Japan (north part of the Sea of Japan and southeastern part of the Pacific Coast of Japan), Sarsiella japonica Hiruta, 1977 and S. misakiensis Kajiyama, 1912, are redescribed. Based on 11 newly obtained COI sequences we construct a preliminary phylogenetic tree, which supports previous hypotheses based on the morphological data, that Eusarsiella Cohen & Kornicker, 1975 is a polyphyletic taxon. With the maps of species distribution provided for each of the three genera, we give an overview of their current zoogeography, and clearly indicate areas that have no data, mostly due to the lack of investigation.

Redescription of Coronatella poppei (Richard, 1897) (Crustacea, Branchiopoda, Chydoridae) and a revision of the genus in Brazil, with descriptions of new taxa.

The description of the genus Coronatella Dybowski & Grochowski, 1894 (Cladocera: Chydoridae: Aloninae) pointed towards the need for a revision of species on a worldwide scale. For the Neotropical region, the main challenge noted was the redescription of Coronatella poppei (Richard, 1897). We redescribed this species and revised populations from Brazil that had previously been assumed to be Alona poppei (= C. poppei). Our results indicate that C. poppei is distributed in the southern part of South America. In Brazil, two other taxa are recognized, Coronatella paulinae sp.nov. and Coronatella serratalhadensis sp.nov., which are morphologically distinguished both from each other and from C. poppei. These species also have different geographic distributions. The Brazilian Coronatella fauna also comprises Coronatella monacantha (Sars, 1901) and a related species, Coronatella undata sp.nov. Our results point towards a previously unknown high diversity of Coronatella in the Neotropical region with several implications for to biogeography of the genus.