Indo-West Pacific species of the genus Stylopoma Levinsen, 1909 (Bryozoa: Cheilostomatida)

Twenty-one species of Stylopoma Levinsen, 1909 are described and illustrated with scanning electron micrographs. S. spongites (Pallas, 1766), the type species, is redescribed from the neotype specimen and included here for completeness. A neotype specimen for S. duboisii (Audouin, 1826) and a lectotype specimen for S. viride (Thornely, 1905) are designated and 14 new species of Stylopoma are described from Indo-West Pacific material in the Recent bryozoan collections of the Natural History Museum, London.     

Morphological aspects of fertilization in Phallusia (Ascidia) nigra (Ascidiacea,Tunicata)

The spermatozoa of Phallusia (Ascidia) nigra have an elongated head (approximately 5 m in length) in which a nucleus and a single mitochondrion are located side by side. There is no midpiece. The apex of the head is wedge-shaped. Acrosomal vesicles (approximately 55–65 nm in diameter) and moderately electron-dense material (MEDM) are present between the plasmalemma and the nuclear membranes in the anterior tip of the head. The MEDM occupies a central position and three or four acrosomal vesicles are seen in a line alongside it. The acrosomal vesicles disappear as the sperm makes contact with the surface of the chorion. Gamete fusion most likely occurs between a small process extending from the peripheral margin of the sperm apex and the egg surface, resulting in incorporation of the sperm into the egg from the anterior region of its head.     

Morphology of the first zoeal stages of eleven Sesarmidae (Crustacea,Brachyura, Thoracotremata) from the Indo-West Pacific,with a summary of familial larval characters

Summary

The first zoeal stages of eleven species of Sesarmidae from the Indo-West Pacific were obtained from ovigerous females. Those of Labuanium scandens, L. rotundatum, L. trapezoideum, L. politum, Metasesarma aubryi, Pseudosesarma crassimanum, Stelgistra stormi, and Sesarmops impressum, are described for the first time, while the first zoeal stages of Clistocoeloma merguiense, Metasesarma obesum and Sesarmops intermedium are re-described. Larval characters of all these species are compared with previously described ones for the family and morphological features are re-evaluated. Minute spines on the telson of the zoeae are described as a new larval character in Sesarmidae and their presence or absence in other grapsoid groups is discussed. The results demonstrate that a recurrent combination of reliable larval characters distinguishes zoeae and megalopae of the examined sesarmids from the rest of the Grapsoidea. This appears to be consistent with recent studies that redefine the Sesarmidae.     

New lucinid bivalves from shallow and deeper water of the Indian and West Pacific Oceans (Mollusca,Bivalvia, Lucinidae)

Four new species and a new genus of lucinid bivalves are described from shallow and deeper waters in the Indian and West Pacific Oceans. The new genus Scabrilucina (subfamily Lucininae) includes the little-known Scabrilucina victorialis (Melvill, 1899) from the Arabian Sea and Scabrilucina vitrea (Deshayes, 1844) from the Andaman Sea as well as a new species Scabrilucina melvilli from the Torres Strait off northeastern Australia. Ferrocina brunei new species (Lucininae) was recovered from 60 m near oil drilling activities off Borneo; its anatomy confirmed the presence of symbiotic bacteria. Two unusual deeper water species of Leucosphaerinae are described, both species included in on-going molecular analyses; Gonimyrtea ferruginea from 400–650 m in the southwest Pacific and Myrtina reflexa from 200–825 m off Zanzibar and Madagascar.     

Ultrastructural Aspects of Spicule Formation in the Solitary Ascidian Herdmania momus (Urochordata,Ascidiacea)

The solitary stolidobranch ascidian Herdmania momus contains numerous calcium carbonate spicules in its tunic and body tissues. The slender body spicules form inside complex sheaths in the body wall and branchial basket, where they remain for the life of the animal. The much smaller tunic spicules form inside the tunic blood vessels and then migrate to the tunic surface, where they become anchored by their spiny base. This paper is an ultrastructural investigation of the formation of the body spicules; the tunic spicules, which apparently form quite differently, will be the focus of a future study. The body spicules are composed of rows of closely packed acicular spines which form completely extracellularly. The spine tips are covered by flattened, highly pseudopodial sclerocytes bound together by tightly interdigitating cell processes. The basal regions of contiguous spines are covered by very thin sclerocyte cell processes. An organic matrix is present within the spines; its exact nature is not clear. A very dense extracellular inter-spine matrix is located between the spine tips and the contiguous basal regions. Presclerocytes within the sheaths between the spicules are probably responsible for formation of the extracellular structures of the sheaths. The presclerocytes appear to aggregate and transform into sclerocytes at the apical end of the spicule. New spines are added at the apical end of the spicule as well as between larger spines. Comparisons are made between body spicule formation in H. momus and skeletogenesis in echinoids.     

The Australian endemic woodwasp genus Austrocyrta Riek (Hymenoptera: Xiphydriidae)

The Australian xiphydriid woodwasp genus Austrocyrta , previously known only from a single species, A. australiensis Riek, is revised. A second distinctive species from Carnarvon National Park in south-central Queensland, A. fasciculata Jennings & Austin, sp. nov., is described, and a key to separate the males of the two species is provided.     

Ovulation in Diplosoma (Tunicata, Ascidiacea, Didemnidae): a light microscopical study

Serial sections of non-ovulated and ovulated eggs of Diplosoma migrans (Tunicata, Ascidiacea, Didemnidae) were compared in order to study the process of ovulation on the level of light microscopy. Fully grown eggs are surrounded by an outer follicular epithelium consisting of cubic cells, and an inner follicular epithelium consisting of flat cells. Egg release is initiated by a fusion of the epidermis with the outer follicular epithelium at the distal pole of the egg. A contraction of the outer follicular epithelium leads to an opening of both epidermis and outer follicular epithelium, and their removal from the egg. The envelope of ovulated eggs corresponds to the former inner follicular epithelium. Mainly due to a contraction of the epidermis, the outer follicular epithelium is shifted into the abdomen and separates from the epidermis. It transforms into a vestigial body (corpus luteum) and finally atrophies. The epidermis flattens and resumes its typical appearance. On the basis of these observations, the results of studies on the closely related Diplosoma listerianum are discussed and questioned. Electronic Publication     

Cupular organs in two species of Corella (Tunicata: Ascidiacea)

Abstract. Simple cupular organs similar to those described in Ciona intestinalis were observed in Corella eumyota. They consist of a macula containing the cell bodies of 20–30 primary sensory neurons whose cilia project into a dome‐ or finger‐shaped structure, the cupula. Rather than being found in the mantle lining as in C. intestinalis, the organs were located on the atrial surface of the branchial sac. The sensory innervation was examined in whole‐mount preparations using anti‐tubulin immunohistochemistry. Sensory neurons in C. eumyota showed no immunoreactivity with antisera raised against gonadotropin‐releasing hormone (GnRH). A novel, elongated sense organ termed the cupular strand was found in Corella inflata. It has the same basic components as the simple type of cupular organ but consists of a single, long structure containing ～1500 sensory cells. Located on the atrial surface of the branchial sac, it extends along the midline of the dorsal fold, from the gonoduct openings almost as far as the brain. Preparations were examined using optical and electron microscopy. Nerves and cilia were visualized by anti‐tubulin immunofluorescence microscopy. It was possible to follow the sensory axons from the macula of the cupular strand to points where they joined branches of the visceral nerve, which enters a nerve root at the back of the brain. In C. inflata the sensory cell bodies and their axons were immunoreactive not only with anti‐tubulin but also with an antiserum raised against Tunicate I GnRH. There was no immunoreactivity, however, with Chicken II and catfish GnRH antisera. All three GnRH antisera labeled the dorsal strand plexus, a structure associated with production of GnRH in its role as a reproductive hormone. We concluded that the GnRH‐like molecule labeled in sensory neurons differs from the form of GnRH found in the dorsal strand plexus, and may have a different function, perhaps in the neural control of ciliary activity. The function of the cupular organs in species of Corella has not yet been investigated physiologically, but by analogy with such structures in other metazoans, cupular organs are probably hydrodynamic sensors registering local disturbances or changes in water flow through the atrial cavity.     

Glycogen and degeneration in the pyloric gland of Dendrodoa grossularia (Ascidiacea,Tunicata)

Summary Degeneration is observed in cells of the pyloric gland of Dendrodoa grossularia in which glycogen storage occurs. The ultrastructure of four phases of the degeneration cycle is described. Natural senescence seems to be the cause of the degeneration. Glycogen storage might be the result of metabolic disturbance, but its presence reveals the importance of glycogen in the function of the organ. The role of the pyloric gland is discussed.     

Historical biogeography and molecular systematics of the Indo-Pacific genus Dascyllus (Teleostei: Pomacentridae)

The phylogeographical and systematic relationships among species in the tropical marine fish genus Dascyllus were inferred using mitochondrial DNA (mtDNA) sequence data. Although our results were generally consistent with previously published phylogenies based on both morphological and mitochondrial data, our broad taxonomic and geographical sampling design revealed novel insights into the phylogenetic history of Dascyllus that had escaped previous notice. These results include: (a) the polyphyletic nature of D. reticulatus mtDNAs, representing two divergent and geographically separated lineages, one shared with D. flavicaudus and the second forming the sister lineage of D. carneus; (b) the paraphyly of D. trimaculatus relative to the closely related D. abisella; and (c) phylogeographical structure within the widespread taxa D. aruanus and D. trimaculatus. Application of a molecular clock permits us to posit a causative role for tectonic and oceanic changes regarding some Dascyllus speciation events. Finally, we mapped body size and the presence or absence of protogynous sex change on the mtDNA tree, and tested published hypotheses regarding determinants of the evolution of mating system and protogyny in the genus. Our data rejected a model based on body size but not one based on phylogenetic inertia. The ability to change sex arose once in the ancestor to the entire genus, and was lost once in the ancestor of the D. trimaculatus complex. For taxa that are as geographically widespread as many Indo-Pacific genera, this study highlights the importance of adequate geographical sampling when inferring patterns of species diversification and life history evolution.     

Biological activity in extracts of ascidians (Tunicata, Ascidiacea) from the northeastern Brazilian coast

Ascidians are marine animals with a great ability to synthesize bioactive substances. This study examined the cytotoxic potential of 10 ascidians found in the coastal waters of Northeast Brazil. Samples of the species Eudistoma vannamei Millar, 1977, Eudistoma sp., Didemnum ligulum Monniot F., 1983, Didemnum psammatodes (Sluiter, 1895), Didemnum sp., Polysyncraton sp., Trididemnum sp., Cystodytes dellechiajei (Della Valle, 1877), Euherdmania sp., and an unidentified species belonging to the Holozoidae family were extracted in methanol 5:1 (v/w). The extracts were tested for cytotoxicity using the brine shrimp lethality assay, sea urchin egg development assay, hemolysis assay, and MTT assay using tumor cell lines. The extract of E. vannamei showed the highest toxicity in brine shrimp (LD₅₀=34.7 μg/ml) and in all tumor cell lines tested, with an IC₅₀ of <2 μg/ml for CEM, 11.2 μg/ml for HL-60, 23.8 μg/ml for B16, and 14.3 μg/ml for HCT-8. In sea urchin eggs, it inhibited the cell cycle progression mainly at the blastula stage (IC₅₀=74.8 μg/ml). The extract of Euherdmania sp. also exhibited some toxicity in these assays, but at a lower potency than that of E. vannamei. The extracts of D. psammatodes and Polysyncraton sp. showed a strong inhibition of the sea urchin egg cell cycle during both phases examined, first cleavage and blastula, with a possible action on the cell microfilament apparatus. The extract of D. ligulum showed selective toxicity toward HCT-8 cells (IC₅₀=35.3 μg/ml). The extract from the Holozoidae was the only one that possessed a hemolytic effect, with an IC₅₀ of 175.2 μg/ml. Further studies are necessary for a better characterization of the active principles of these extracts and a possible elucidation of the mechanisms of action.     

Morpho-functional characterization of haemocytes of the compound ascidian Botrylloides leachi (Tunicata, Ascidiacea)

A morpho‐functional study of the colonial ascidian Botrylloides leachi haemocytes was carried out to propose their classification, relationships and specializations. This characterization was obtained by (i) investigations of both living and aldehyde‐fixed cells by light and electron microscopy; (ii) cytochemical and cytoenzymatic assays; (iii) lectin‐affinity assays; (iv) phagocytosis and haemagglutination assays; and (v) anti‐CD34 immunocytochemical assay for vertebrate haematopoietic stem cells. Results indicate that the haemoblast is a circulating stem cell and there are at least five haemocyte differentiation pathways, the last two of which have never been described in botryllids: (i) phagocytic line (hyaline amoebocytes and macrophage‐like cells) share ultrastructural features, the same hydrolytic enzymes and WGA lectin binding, and are involved in yeast phagocytosis and erythrocyte rosette formation; (ii) cytotoxic line (granular amoebocytes and morula cells) with vacuoles containing oxidative enzymes and polyphenolic substrates; (iii) vacuolated cell line (pigment cells and nephrocytes) involved in catabolite storage; (iv) compartment cell line (compartment amoebocytes and compartment cells) able to agglutinate erythrocytes and characterized by vacuoles with a moderately electron‐dense content, positive to arylsulphatase activity and binding DBA, UEA‐I, HPA lectins; and (v) granular cell line includes trophic cells, able to infiltrate the gut epithelium, showing a cytoplasm filled of PAS‐positive vacuoles with arylsulphatase, chloroacetylesterase and β‐glucuronidase activities.     

An experimental test of colonization ability in the potentially invasive Didemnum perlucidum (Tunicata, Ascidiacea)

Exotic species invasions are one of the greatest threats to marine systems and ascidians have many characteristics that favor transport, colonization and establishment into new regions. Didemnum perlucidum is a widespread species that has been introduced into tropical ports around the world. Here we examine the colonizing ability of D. perlucidum by experimental use of artificial plates in a shellfish culture. The experiment comprised paired plates for colonization (bare and occupied) in 16 monthly replicates. Recruitment and space occupation were compared between bare and occupied plates and an estimation of reproductive effort was based on the number of larvae produced in each of ten colonies collected on the culture structures. D. perlucidum reproduced continuously but greatest reproduction occurred between December 2006 and May 2007. While recruitment was somewhat greater (number of new colonies) on bare plates, this species can colonize already occupied substrates and, surprisingly, colony area was always similar between treatments. Thus, while fewer colonies formed on occupied plates, once formed, colonies grew at rates similar to those on clean plates. Thus, D. perlucidum colonizes substrates very efficiently, especially when unoccupied space is available.     

A morphological and immunohistochemical study of programmed cell death in Botryllus schlosseri (Tunicata,Ascidiacea)

The blastogenic cycle of the colonial ascidian Botryllus schlosseri concludes in a phase of selective cell and zooid death called takeover. Every week, all asexually derived parental zooids synchronously regress over a 30-h period and are replaced by a new generation. Here we document the sequential ultrastructural changes which accompany cell death during zooid degeneration. The principal mode of visceral cell death during takeover occurred by apoptosis, the majority of cells condensing and fragmenting into multiple membrane-bounded apoptotic bodies. Cytoplasmic organelles (mitochondria, basal bodies, striated rootlets) within apoptotic bodies retained ultrastructural integrity. Dying cells and fragments were then swiftly ingested by specialized blood macrophages or intraepithelial phagocytes and subsequently underwent secondary necrotic lysis. Certain organs (stomach, intestine) displayed a combination of necrotic and apoptotic changes. Lastly, the stomach, which demonstrated some of the earliest regressive changes, exhibited intense cytoplasmic immunostaining with a monoclonal antibody to ubiquitin at the onset of takeover. Affinity-purified rabbit antiserum against sodium dodecyl sulfate-denatured ubiquitin detected a characteristic 8.6-kDa mono-ubiquitin band by Western blot analysis. Collectively, these findings raise the possibility that cell death during takeover is a dynamic process which requires active participation of cells in their own destruction.     

A new marine cyclopoid copepod of the genus Neocyclops (Cyclopidae,Halicyclopinae) from Korea

A new cyclopoid species of the genus Neocyclops Gurney, 1927 is described. Type specimens were collected from a beach on south-western coast of the Korean Peninsula by rinsing intertidal coarse sandy sediments. Neocyclops hoonsooi sp. n. is most characteristic in showing the conspicuous chitinized transverse ridges originating from the medial margins of the coxae of all swimming legs. The new species is most similar to Neocyclops vicinus, described from the Brazilian coast, and Neocyclops petkovskii, from Australia. All three species share a large body size (more than 750 µm long), the presence of an exopodal seta on the antenna, two setae on the mandibular palp, the same seta/spine armature on the third endopodal segment of leg 3 (3 setae + 3 spines), and the fairly long inner distal spine on the third endopodal segment of the female leg 4. However, Neocyclops hoonsooi sp. n. differs from both species by the much shorter caudal rami (less than 1.7 times as long as wide) and the shorter dorsal caudal seta VII. Furthermore, Neocyclops hoonsooi is clearly distinguished from Neocyclops vicinus by the 10-segmented antennule (vs 12 segments in Neocyclops vicinus), and from Neocyclops petkovskii by the elongate inner distal spine on leg 5 exopod and the 3-segmented leg 5 in male (vs 4-segmented in Neocyclops petkovskii). A tabular comparison of characters separating Neocyclops hoonsooi from its closest allies and a key to Neocyclops species from the Indo-Pacific Ocean are provided. This is the first record of the genus Neocyclops from the northern Pacific.