The ultrastructure of resurrection: Post-diapause development in an Antarctic freshwater copepod.

The copepod, Boeckella poppei, is broadly distributed in Antarctic and subantarctic maritime lakes threatened by climate change and anthropogenic chemicals. Unfortunately, comparatively little is known about freshwater zooplankton in lakes influenced by the Southern Ocean. In order to predict the impact of climate change and chemicals on freshwater species like B. poppei, it is necessary to understand the nature of their most resilient life stages. Embryos of B. poppei survive up to two centuries in a resilient dormant state, but no published studies evaluate the encapsulating wall that protects theses embryos or their development after dormancy. This study fills that knowledge gap by using microscopy to examine development and the encapsulating wall in B. poppei embryos from Antarctica. The encapsulating wall of B. poppei is comprised of three layers that appear to be conserved among crustacean zooplankton, but emergence and hatching are uniquely delayed until the nauplius is fully formed in this species. Diapause embryos in Antarctic sediments appear to be in a partially syncytial mid-gastrula stage. The number of nuclei quadruples between the end of diapause and hatching. Approximately 75% of yolk platelets are completely consumed during the same time period. However, some yolk platelets are left completely intact at the time of hatching. Preservation of complete yolk platelets suggests an all-or-none biochemical process for activating yolk consumption that is inactivated during dormancy to preserve yolk for post-dormancy development. The implications of these and additional ultrastructural features are discussed in the context of anthropogenic influence and the natural environment.

Nanoscale trace metal imprinting of biocalcification of planktic foraminifers by Toba's super-eruption.

Bioactive metal releases in ocean surface water, such as those by ash falls during volcanic super-eruptions, might have a potentially toxic impact on biocalcifier planktic microorganisms. Nano-XRF imaging with the cutting-edge synchrotron hard X-ray nano-analysis ID16B beamline (ESRF) revealed for the first time a specific Zn- and Mn-rich banding pattern in the test walls of Globorotalia menardii planktic foraminifers extracted from the Young Toba Tuff layer, and thus contemporaneous with Toba's super-eruption, 74,000 years ago. The intra-test correlation of Zn and Mn patterns at the nanoscale with the layered calcareous microarchitecture, indicates that the incorporation of these metals is syngenetic to the wall growth. The preferential Mn and Zn sequestration within the incipient stages of chamber formation suggests a selective incorporation mechanism providing a resilience strategy to metal pollution in the test building of planktic foraminifers.

Guanine crystals regulated by chitin-based honeycomb frameworks for tunable structural colors of sapphirinid copepod, Sapphirina nigromaculata.

Sapphirinid copepods, which are marine zooplankton, exhibit tunable structural colors originating from a layered structure of guanine crystal plates. In the present study, the coloring portion of adult male of a sapphirinid copepod, Sapphirina nigromaculata, under the dorsal body surface was characterized to clarify the regulation and actuation mechanism of the layered guanine crystals for spectral control. The coloring portions are separated into small domains 70-100 µm wide consisting of an ordered array of stacked hexagonal plates ~1.5 µm wide and ~80 nm thick. We found the presence of chitin-based honeycomb frameworks that are composed of flat compartments regulating the guanine crystal plates. The structural color is deduced to be tuned from blue to achromatic via yellow and purple by changing the interplate distance according to vital observation and optical simulation using a photonic array model. The framework structures are essential for the organization and actuation of the particular photonic arrays for the exhibition of the tunable structural color.

Ultrastructure of Meelsvirus: A nuclear virus of arrow worms (phylum Chaetognatha) producing giant "tailed" virions.

Most known giant viruses, i.e., viruses producing giant virions, parasitize amoebae and other unicellular eukaryotes. Although they vary in the level of dependence on host nuclear functions, their virions self-assemble in the host cell's cytoplasm. Here we report the discovery of a new prototype of giant virus infecting epidermal cells of the marine arrow worm Adhesisagitta hispida. Its 1.25 µm-long virions self-assemble and accumulate in the host cell's nucleus. Conventional transmission electron microscopy reveals that the virions have a unique bipartite structure. An ovoid nucleocapsid, situated in a broad "head" end of the virion is surrounded by a thin envelope. The latter extends away from the head to form a voluminous conical "tail" filled with electron-dense extracapsidular material. The 31nm-thick capsid wall has a distinctive substructure resulting from a patterned arrangement of subunits; it bears no ultrastructural resemblance to the virion walls of other known giant viruses. The envelope self-assembles coincident with the capsid and remotely from all host membranes. We postulate that transmission to new hosts occurs by rupture of protruding virion-filled nuclei when infected arrow worms mate. Future genomic work is needed to determine the phylogenetic position of this new virus, which we have provisionally named Meelsvirus.

Morphology and molecular phylogeny of epizoic araphid diatoms on marine zooplankton, including Pseudofalcula hyalina gen. & comb. nov. (Fragilariophyceae, Bacillariophyta).

Some diatoms are able to colonize as epibionts on their potential zooplankton predators. Here, we report Pseudohimantidium pacificum living on the copepod Corycaeus giesbrechti and as a new finding on Oithona nana, Protoraphis atlantica living on the copepod Pontellopsis brevis, Protoraphis hustedtiana on the cypris larvae of barnacles, and Falcula hyalina on the copepod Acartia lilljeborgii. The epizoic diatoms were able to grow as free-living forms under culture conditions. Pseudohimantidium pacificum and P. atlantica appeared as the most derived species from their benthic diatom ancestors. The mucilage pad or stalk of the strains of these species showed important morphological distinction when compared with their epizoic forms. Barnacle larvae explore benthic habitats before settlement, and epibiosis on them is an example where P. hustedtiana profits from the host behavior for dispersal of its benthic populations. Molecular phylogenies based on the SSU rRNA and RuBisCO large subunit (rbcL) gene sequences revealed F. hyalina as an independent lineage within the Fragilariales (Tabularia, Catacombas, and others), consistent with its morphological distinction in the low number of rows (≤6) in the ocellulimbus, among other features. We propose the transfer of F. hyalina to the genus Pseudofalcula gen. nov. Molecular phylogeny suggests a single order for the members of the Cyclophorales and the Protoraphidales, and that the epibioses of araphid diatoms on marine zooplankton have been independently acquired several times. These clades are constituted of both epizoic and epiphytic/epilithic forms that evidence a recent acquisition of the epizoic modus vivendi.

Predatory flying squids are detritivores during their early planktonic life.

Cephalopods are primarily active predators throughout life. Flying squids (family Ommastrephidae) represents the most widely distributed and ecologically important family of cephalopods. While the diets of adult flying squids have been extensively studied, the first feeding diet of early paralarvae remains a mystery. The morphology of this ontogenetic stage notably differs from other cephalopod paralarvae, suggesting a different feeding strategy. Here, a combination of Laser Capture Microdissection (LCM) and DNA metabarcoding of wild-collected paralarvae gut contents for eukaryotic 18S v9 and prokaryotic 16S rRNA was applied, covering almost every life domain. The gut contents were mainly composed by fungus, plants, algae and animals of marine and terrestrial origin, as well as eukaryotic and prokaryotic microorganisms commonly found in fecal pellets and particulate organic matter. This assemblage of gut contents is consistent with a diet based on detritus. The ontogenetic shift of diet from detritivore suspension feeding to active predation represents a unique life strategy among cephalopods and allows ommastrephid squids to take advantage of an almost ubiquitous and accessible food resource during their early stages. LCM was successfully applied for the first time to tiny, wild-collected marine organisms, proving its utility in combination with DNA metabarcoding for dietary studies.

Two new species of the genus Monstrillopsis Sars, 1921 (Copepoda: Monstrilloida: Monstrillidae) from South Korea.

Monstrilloid copepods of the genus Monstrillopsis Sars, 1921 are recorded from Korea, as well as from the northwestern Pacific, for the first time. Specimens were obtained using a light trap set overnight at piers or seawalls in small harbors. Males of two new species were found and are described herein with remarks on their affinities and detailed drawings and SEM photographs of their morphological features. Monstrillopsis longilobata sp. nov. is allied to M. chathamensis Suárez-Morales & Morales-Ramírez, 2009 and M. sarsi Isaac, 1974 in having the character combination of a long, claw-like apical spinous process on the male antennule and elongate genital lappets. It differs from them in having conspicuous transverse striations on the dorsal surface of the cephalothorax and extremely long male genital lappets, which extend far beyond the anterior margins of the caudal rami. Monstrillopsis coreensis sp. nov. also has a claw-like apical spinous process of the male antennule and conspicuous transverse striations on the dorsal surface of the cephalothorax, but it differs from M. longilobata in having a much longer and stronger apical spine of the antennule and a relatively shorter and undulating genital apparatus.

Nanostructures and Monolayers of Spheres Reduce Surface Reflections in Hyperiid Amphipods.

Transparent zooplankton and nekton are often nearly invisible when viewed under ambient light in the pelagic zone [1-3]. However, in this environment, where the light field is directional (and thus likely to cause reflections), and under the bioluminescent searchlights of potential predators, animals may be revealed by reflections from their body surface [4-7]. We investigated the cuticle surfaces of seven species of hyperiids (Crustacea; Amphipoda) using scanning electron microscopy and found two undocumented features that may reduce reflectance. We found that the legs of Cystisoma spp. (n = 5) are covered with an ordered array of nanoprotuberances 200 ± 20 nm SD in height that function optically as a gradient refractive index material [6, 8, 9]. Additionally, we observed that Cystisoma and six other species of hyperiids are covered with a monolayer of homogenous spheres (diameters ranging from 52 ± 7 nm SD on Cystisoma spp. to 320 ± 15 nm SD on Phronima spp.). Optical modeling using effective medium theory and transfer matrix methods demonstrated that both the nanoprotuberances and the monolayers reduce reflectance by as much as 100-fold, depending on the wavelength and angle of the incident light and the thickness of the gradient layer. Even though we only consider surface reflectance and not internal light scattering, our study demonstrates that these nanoprotuberances and spheres can improve crypsis in a featureless habitat where the smallest reflection can render an animal vulnerable to visual predation.

A unique apposition compound eye in the mesopelagic hyperiid amphipod Paraphronima gracilis.

The mesopelagic habitat is a vast space that lacks physical landmarks and is structured by depth, light penetration, and horizontal currents. Solar illumination is visible in the upper 1,000 m of the ocean, becoming dimmer and spectrally filtered with depth-generating a nearly monochromatic blue light field. The struggle to perceive dim downwelling light and bioluminescent sources and the need to remain unseen generate contrasting selective pressures on the eyes of mesopelagic inhabitants. Hyperiid amphipods are cosmopolitan members of the mesopelagic fauna with at least ten different eye configurations across the family-ranging from absent eyes in deep-living species to four enlarged eyes in mesopelagic individuals. The hyperiid amphipod Paraphronima gracilis has a pair of bi-lobed apposition compound eyes, each with a large upward-looking portion and a small lateral-looking portion. The most unusual feature of the P. gracilis eye is that its upward-looking portion is resolved into a discontinuous retina with 12 distinct groups, each serving one transverse row of continuously spaced facets. On the basis of eye morphology, we estimated spatial acuity (2.5° ± 0.11°, SEM; n = 25) and optical sensitivity (30 ± 3.4 µm(2) ⋅ sr, SEM; n = 25). Microspectrophotometry showed that spectral sensitivity of the eye peaked at 516 nm (±3.9 nm, SEM; n = 6), significantly offset from the peak of downwelling irradiance in the mesopelagic realm (480 nm). Modeling of spatial summation within the linear retinal groups showed that it boosts sensitivity with less cost to spatial acuity than more typical configurations.

Phytoplankton cell size: intra- and interspecific effects of warming and grazing.

Decreasing body size has been suggested as the third universal biological response to global warming after latitudinal/altitudinal range shifts and shifts in phenology. Size shifts in a community can be the composite result of intraspecific size shifts and of shifts between differently sized species. Metabolic explanations for the size shifts dominate in the literature but top down effects, i.e. intensified size-selective consumption at higher temperatures, have been proposed as alternative explanation. Therefore, we performed phytoplankton experiments with a factorial combination of warming and consumer type (protist feeding mainly on small algae vs. copepods mainly feeding on large algae). Natural phytoplankton was exposed to 3 (1(st) experiment) or 4 (2(nd) experiment) temperature levels and 3 (1(st) experiment: nano-, microzooplankton, copepods) or 2 (2(nd) experiment: microzooplankton, copepods) types of consumers. Size shifts of individual phytoplankton species and community mean size were analyzed. Both, mean cell size of most of the individual species and mean community cell size decreased with temperature under all grazing regimes. Grazing by copepods caused an additional reduction in cell size. Our results reject the hypothesis, that intensified size selective consumption at higher temperature would be the dominant explanation of decreasing body size. In this case, the size reduction would have taken place only in the copepod treatments but not in the treatments with protist grazing (nano- and microzooplankton).

Filtration of submicrometer particles by pelagic tunicates.

Salps are common in oceanic waters and have higher per-individual filtration rates than any other zooplankton filter feeder. Although salps are centimeters in length, feeding via particle capture occurs on a fine, mucous mesh (fiber diameter d approximately 0.1 microm) at low velocity (U = 1.6 +/- 0.6 cmxs(-1), mean +/- SD) and is thus a low Reynolds-number (Re approximately 10(-3)) process. In contrast to the current view that particle encounter is dictated by simple sieving of particles larger than the mesh spacing, a low-Re mathematical model of encounter rates by the salp feeding apparatus for realistic oceanic particle-size distributions shows that submicron particles, due to their higher abundances, are encountered at higher rates (particles per time) than larger particles. Data from feeding experiments with 0.5-, 1-, and 3-microm diameter polystyrene spheres corroborate these findings. Although particles larger than 1 microm (e.g., flagellates, small diatoms) represent a larger carbon pool, smaller particles in the 0.1- to 1-microm range (e.g., bacteria, Prochlorococcus) may be more quickly digestible because they present more surface area, and we find that particles smaller than the mesh size (1.4 microm) can fully satisfy salp energetic needs. Furthermore, by packaging submicrometer particles into rapidly sinking fecal pellets, pelagic tunicates can substantially change particle-size spectra and increase downward fluxes in the ocean.

Induced metamorphosis in crustacean y-larvae: towards a solution to a 100-year-old riddle.

BACKGROUND: The y-larva, a crustacean larval type first identified more than 100 years ago, has been found in marine plankton samples collected in the arctic, temperate and tropical regions of all oceans. The great species diversity found among y-larvae (we have identified more than 40 species at our study site alone) indicates that the adult organism may play a significant ecological role. However, despite intense efforts, the adult y-organism has never been identified, and nothing is therefore known about its biology. RESULTS: We have successfully and repeatedly induced metamorphosis of y-larvae into a novel, highly reduced juvenile stage by applying the crustacean molting hormone 20-HE. The new stage is slug-like, unsegmented and lacks both limbs and almost all other traits normally characterizing arthropods, but it is capable of vigorous peristaltic motions. CONCLUSION: From our observations on live and preserved material we conclude that adult Facetotecta are endoparasitic in still to be identified marine hosts and with a juvenile stage that represents a remarkable convergence to that seen in parasitic barnacles (Crustacea Cirripedia Rhizocephala). From the distribution and abundance of facetotectan y-larvae in the world's oceans we furthermore suggest that these parasites are widespread and could play an important role in the marine environment.

New records of Monstrilloida Sars, 1901 (Crustacea, Copepoda) on the Brazilian northeastern coast / Novas ocorrências para Monstrilloida Sars, 1901 (Crustacea, Copepoda) na costa Nordeste do Brasil

During a series of zooplankton surveys carried out from 2001 through 2005 off the coast of the state of Bahia, Brazil, 98 individuals of monstrilloid copepods were collected. These belong to five species (Monstrilla grandis, Cymbasoma cf. longispinosum, Cymbasoma cf. rigidum, Cymbasoma gracilis, and Cymbasoma quadridens). The first three are recorded for the first time in the Bahia coastal region. The geographical range of C. quadridens is expanded to the Brazilian northeastern coast. The results presented herein increase to nine the number of nominal species of Monstrilloida known from off Bahia; the environmental diversity of Caravelas Channel with highly productive areas and coral reef zones harbor an abundant and diverse monstrilloid fauna that should be surveyed in more detail.

Durante uma série de amostragens de zooplâncton, realizadas de 2001 a 2005, na costa do estado de Bahia, Brasil, 98 exemplares de copépodes Monstrilloida foram coletados. Os exemplares pertencem a cinco espécies (Monstrilla grandis, Cymbasoma cf. longispinosum, Cymbasoma cf. rigidum, Cymbasoma gracilis and Cymbasoma quadridens). As primeiras três espécies foram coletadas pela primeira vez na região costeira do estado da Bahia. A distribuição geográfica de C. quadridens foi expandida para a costa nordeste brasileira. Os resultados apresentados aumentam a nove o número de espécies Monstrilloida conhecidas para a costa da Bahia. A diversidade ambiental do Canal de Caravelas, com áreas altamente produtivas e a proximidade de uma região de recifes de coral, abriga uma abundante e diversa fauna de Monstrilloida, que deve ser examinada mais detalhadamente.

Docosahexaenoic acid production and lipid-body formation in Schizochytrium limacinum SR21.

Schizochytrium limacinum SR21, a thraustochytrid (Labyrinturomycota), is a heterotrophic marine microorganism. SR21 has attracted recent attention because of the production of docosahexaenoic acid (DHA). We obtained highly concentrated SR21 zoospores and successfully observed synchronous growth. We investigated changes of lipid content and fatty acid composition during the growth. The morphological features of the lipid bodies were also described via fluorescent and electron microscopy. The cells developed quickly after zoospore settlement. Lipid bodies developed in accordance with an increase in lipid content during the 8-h synchronous growth. The total lipid was composed mainly of triacylglycerol, sterol esters, and phosphatidylcholine. The proportion of neutral lipids (triacylglycerol and sterol esters) in the total lipid was fairly constant during growth. The fatty acid composition of neutral lipids, primary components of the lipid body, and phospholipids, primary components of the cell membranes, was nearly unchanged during the synchronous growth. However, the DHA content of the phospholipids decreased drastically after a 10-day culture. Electron micrographs prepared using a high-pressure freeze substitution technique revealed a fine structure of light- and dark-staining bands inside the lipid bodies in many stages of the cells.

Urotricha psenneri n. sp. and Amphileptus piger (Vuxanovici, 1962) n. comb., two planktonic ciliates (Protozoa, Ciliophora) from an oligotrophic lake in Austria.

Two euplanktonic ciliates, Urotricha psenneri n. sp. (Prostomatida) and Amphileptus piger (Vuxanovici, 1962) n. comb. (Pleurostomatida), were discovered in the surface plankton of the oligotrophic Lake Traunsee in Austria. Their morphology and infraciliature were studied in live cells as well as in specimens impregnated with protargol and silver nitrate. Urotricha psenneri is a small urotrichid, less than 50 microm length and with a single caudal cilium. It is unique in having (i) a massive oral basket projecting as a conspicuous bulge with cylindrical microfibrillar annulus and (ii) a curved brosse row 1 in the broad, barren circumoral area. Amphileptus piger (Vuxanovici, 1962) is about 55 x 13 microm in vivo, has two macronuclear nodules with a single micronucleus in between in the posterior body half, has a single contractile vacuole with a terminal excretory pore, and few, but thick and thus highly conspicuous extrusomes. The amphileptid ciliary pattern (spica) is difficult to recognise due to the widely spaced basal bodies.

Attachment of Vibrio cholerae serogroup O1 to zooplankton and phytoplankton of Bangladesh waters.

Vibrio cholerae serogroup O1, the causative agent of cholera, is capable of surviving in aquatic environments for extended periods and is considered an autochthonous species in estuarine and brackish waters. These environments contain numerous elements that may affect its ecology. The studies reported here examined physical interactions between V. cholerae O1 and natural plankton populations of a geographical region in Bangladesh where cholera is an endemic disease. Results showed that four of five clinical V. cholerae O1 strains and endogenous bacterial flora were attached preferentially to zooplankton molts (exuviae) rather than to whole specimens. One strain attached in approximately equal numbers to both exuviae and whole specimens. V. cholerae O1 also attached to several phytoplankton species. The results show that V. cholerae O1 can bind to diverse plankton species collected from an area where cholera is an endemic disease, with potentially significant effects on its ecology.

A method for cytogenetic and cytological examination of small shelled larvae of bivalves and other zooplankton.

Studies on chromosomes and nuclei of very small bivalve larvae have been impeded by the veliger shell. It has been determined that the alcohol:acetic acid fixative commonly used in cytogenetic techniques can be made to act as a decalcifying agent upon repeated heating. In addition, transfer of formalin fixed shelled specimens, routinely used as marine bioassay organisms, into ethyl alcohol:acetic acid (3:1) fixative also yields clear cells for cytological examination of decalcified but otherwise intact oyster larvae and other zooplankton. Identification of cell type, such as germ-line primordia, in, for example, reproductive and ploidy level studies, and observations on the presence of bacteria can be made from the preparations. Material can be examined up to at least a year after preservation. The method is evaluated and its modifications are discussed.

Movement generated by interactions between the dense material at the ends of microtubles and non-actin-containing microfilaments in Sticholonche zanclea.

Axopods of the planktonic protozoan, Sticholonche, are used as oars to propel the organism through seawater. Within each axopod is an orgainzed array of microtubules which inserts into a dense material that assumes the form of the head of a hip joint. This material, in turn, articulates on the surface of the nucleus. Microfilaments, 20-30 A in diameter, connect the dense material associated with the microtubules to the surface of the nucleus, and they move the axopod by their contractions. The active phase of the movement may take as little as about 0.04 s and the recovery phase may take between 0.2 and 0.4 s. The microfilaments are not actin, as based on: (a) their small diameter, (b) the lack of decoration with heavy meromyosin, and (c) their ability to coil, spiral or fold during contraction. By the use of Thorotrast, we were able to demonstrate that the cell surface is deeply infolded, extending all the way to the hip joint. Here, there is a specialized membrane system that resembles the diad in skeletal muscle. From cytochemical tests and the use of ionophores and chelators, there is some evidence that the motile process may be controlled by calcium. This study demonstrates that, in at least one system, microtubules can be moves by contractile microfilaments attached to the dense material at there tips.

Polymorphism in the rotifer Asplanchna sieboldi. Fine structure of saccate, cruciform and campanulate females.

The morphology of saccate, cruciform, and campanulate females of Asplanchna sieboldi was analyzed by light and electron microscopy. No major qualitative variations were observed in the ultrastructure of the hypodermis, gastric gland and yolk gland of females from the different morphotypes. Campanulate females did, however, possess larger secretory granules in their gastric gland as well as larger and more numerous nuclei in their gastric and yolk glands than both saccate and cruciform females. The structure of the secretory elements in the gastric gland varied between the two clones investigated.

Gametogenesis in planktonic foraminifera.

Gametogenesis in Globigerinella aequilateralis and Globigerinoides sacculifer in culture is preceded by sinking of the organism and loss of its spines. Hundreds of thousands of flagellated gametes, about 5 micrometers in diameter, are produced within the parent shell and released within a period of 13 hours.