Egg capsules, eggs and embryos of the southwestern Atlantic gastropod Coronium coronatum (Gastropoda: Muricidae)

The egg capsules, eggs and embryos of the muricid gastropodCoronium coronatum are described for the first time. Capsulesare sessile, bulliform, semi-circular, with a plug in the dorsalcenter. Sutures split the capsule into two asymmetrical halves.Recently laid capsules showed the presence of 3639 (n = 2) uncleavednurse eggs with a diameter of 180–210 µm (mean= 197.4 ± 8.9). The number of early embryos was 9–11.The size of the embryos was 320 x 320 to 820–880 µm.Nine pre-hatching embryos of 3.94 mm (n = 8, SD = 0.32)were found inside the older capsule. SEM illustrations of embryosand radulae are provided. Comparison of shell and radula ofembryos with the protoconch and radulae of adults of C. coronatumrevealed that the capsule belongs to this species. (Received 18 March 2006; accepted 10 October 2006)     

Embryonic and larval development of Ensis arcuatus (Jeffreys, 1865) (Bivalvia: Pharidae)

The razor clam Ensis arcuatus (Jeffreys, 1865) is distributedfrom Norway to Spain and along the British coast, where it livesburied in sand in low intertidal and subtidal areas. This workis the first study to research the embryology and larval developmentof this species of razor clam, using light and scanning electronmicroscopy. A new method, consisting of changing water levelsusing tide simulations with brief dry periods, was developedto induce spawning in this species. The blastula was the firstmotile stage and in the gastrula stage the vitelline coat waslost. The shell field appeared in the late gastrula. The trochophoredeveloped by about 19 h post-fertilization (hpf) (19°C).At 30 hpf the D-shaped larva showed a developed digestive systemconsisting of a mouth, a foregut, a digestive gland followedby an intestine and an anus. Larvae spontaneously settled after20 days at a length of 378 µm. (Received 5 December 2006; accepted 19 November 2007)     

PREDATOR-PREY INTERACTIONS BETWEEN LEPSIELLA (BEDEVA) PAIVAE (GASTROPODA: MURICIDAE) AND KATELYSIA SCALARINA (BIVALVIA: VENERIDAE) IN PRINCESS ROYAL HARBOUR, WESTERN AUSTRALIA

On a sandy beach at Shoal Bay in Princess Royal Harbour, Albany,southwestern Western Australia, lives a small muricid gastropodthat feeds virtually monotonically on the overwhelmingly dominantresident bivalve Katelysia scalarina. Lepsiella paivae livesburied in the sand and attacks its prey within it. Because ofits small size (<13 mm shell height), bivalve prey isalso small and this study demonstrates a preference for K. scalarinaof 5 mm shell length, i.e. juveniles. Laboratory experimentsalso suggested a possible preference for attack of the rightvalve. Lepsiella paivae can and does, however, attack largerprey (up to 15 mm shell length), but cannot consume themcompletely. A second visit to Princess Royal Harbour in theAustral winter, when there was no juvenile K. scalarina present,showed L. paivae to be attacking at the sand surface, also bydrilling, the small (<4 mm) gastropod Hydrococcus brazieri(Hydrococcidae). SEM studies of experimentally determined drillholes of L. paivae show them to be of variable form, some straightsided, others bevelled (like a naticid) and <500 µmin diameter. On this sheltered Southern Ocean beach, therefore,L. paivae has specialized to attack juvenile bivalves by burrowingafter them. It can, however, attack other species opportunisticallyon the sand surface when seasonally favoured juvenile bivalveprey are not present. (Received 8 January 2005; accepted 16 March 2005)     

Predation of cockles (Cerastoderma edule) by the whelk (Buccinum undatum) under laboratory conditions

The feeding rate and behaviour of whelks (Buccinum undatum)offered cockles (Cerastoderma edule) in laboratory experimentswere examined. When presented with cockles in a range of sizes(10–40 mm), 14 B. undatum (34.6–88.3 mm),held individually in aquaria, consumed a wide size range ofcockles. Small whelks (<40 mm) consumed cockles (<23 mm),whereas large whelks, (>60 mm) ate a greater numberof larger cockles (>30 mm) and a wider size range ofcockles (12–40 mm) than smaller whelks. The majority(90%) of the shells of the predated cockles were undamaged andthe few (<10%) that were damaged showed only slight abrasionsto the anterior and posterior shell margin. Filmed observationsof B. undatum feeding on C. edule showed a method of attackthat has not previously been reported and involved the use ofthe whelk's foot to asphyxiate the cockle or to pull the shellvalves apart. No filmed evidence was found for the previouslyreported shell ‘wedging’ technique for prising openthe closed shell valves of C. edule, although 10% of the shellsof consumed cockles in feeding experiments had damaged shellmargins. (Received 4 April 2007; accepted 30 June 2007)     

Temporal variability in copepod fecundity during two different spring bloom periods in coastal waters off Plymouth (SW England)

The temporal variation in egg production of the planktonic copepodsCalanus helgolandicus, Temora longicornis and Pseudocalanuselongatus was studied during two different spring bloom periodsin 1989 and 1990 by weekly sampling at two permanent stationsin coastal waters off Plymouth (SW England). Copepod egg productionwas estimated in situ by incubating individual adult femalesin filtered seawater for 24 h (72 h until hatching for P.elongatus)at the field surface temperature. The relationship between copepodegg production rates and: (i) chlorophyll a concentration (totaland >10 µm size fraction), (ii) temperature and (iii)initial copepod gut pigment content was investigated. The springbloom periods were very different in both years, with the occurrenceof a Phaeocystis sp. bloom in 1990, which negatively affectedthe feeding and fecundity of copepods. Egg production ratesin spring 1989 were significantly correlated with chlorophylla concentration (particularly with the >10 µm fraction),field temperature and copepod gut pigment contents. In spring1990, egg production rates were also correlated with copepodgut pigment contents, but no significant correlations were obtainedwith temperature or with chlorophyll a concentration, as a consequenceof the lower egg production rates obtained during the Phaeocystissp. dominance period. These results show that food availabilityis the factor which mainly affects the fecundity of neriticcopepods in short time periods.     

Somatic Embryos and Plantlets from an Ancient Clone of the Grapevine (cv. Cabernet-Sauvignon) by Apomixis in vitro

Plantlets were produced by induction of nucellar embryony (apomixis)in isolated unfertilized ovules of a non-apomictic plant—thegrapevine (Vitis vinifera L. cv. Cabernet-Sauvignon). Ovulesgrown in liquid culture with benzyladenine (5–10 µM)and ß-naphthoxyacetic acid (5–25 µM) formeda nucellar callus which gave rise to somatic embryos. For growthof plantlets the embryos were transferred to an agar mediumcontaining gibberellic acid (1 µM) and 2-isopentenyladenine(5 µM). This is the first report of somatic embryo formationin a cultivar of a temperate fruit.     

Grazing of echinoderm larvae (Paracentrotus lividus and Arbacia lixula) on naturally occurring particulate matter

Larvae (72 hr old) of P. lividus and A. lixula grazed on varioussuspensions of natural particulate matter with a size rangeof 2 to 30 microns, and on two species of algae (Phaeodactylumtricor-nutum and Nitzschia sp.) — Larvae graze most in the size range where the particleconcentration is highest. — If larvae deplete certain size categories of particlesthey then graze other size ranges in which the concentrationis still high. — The grazing rate of the two species varied between 988and 91.949 µm³ per pluteus per hour. — For A. lixula larvae the grazing rate increases withincreasing temperature to a maximum at 22°C.     

Embryos and Plantlets from Cultured Anthers of Hybrid Grapevines

Embryos and plantlets were produced in large numbers from callusformed by cultured anthers of hybrid grapevines (Vitis viniferax Vitis rupestris). Anthers of Vitis vinifera produced smallamounts of callus or failed to grow in vitro. For embryo formationanthers containing uninucleate microspores were chilled (4 °C)for 72 h before culture with Nitsch medium containing 2, 4-D(5µM) and benzyladenine (1 µM). Highest yields ofembryos were with anthers cultured in darkness. For productionof normal plantlets embryos required chilling (4 °C) for2 weeks. Unchilled embryos produced mainly abnormal plantlets.Chilling was effective in promoting plantlet growth when appliedat any stage of embryogeny. In grapes ability to produce plantlets from cultured anthersis a genetically-determined trait and maleness, as distinctfrom hermaphroditism, may be a predisposing factor. Callus derivedfrom anthers contained both haploid and diploid cells but allplantlets produced so far are diploid. The genetic constitutionof plantlets, whether they are diploids of somatic origin ordiploids from spontaneously doubled haploid cells, is not yetknown and is being determined by standard genetic methods.     

GROWTH AND REPRODUCTION OF THE MINUTE LAND SNAIL PUNCTUM PYGMAEUM (DRAPARNAUD)

Reproductive characters and growth of Punctum pygmaeum wereinvestigated under laboratory conditions. Eggs were laid singlyat intervals of 2–8 days. Egg size averaged 0.41 by 0.50mm. Due to a variable period of egg retention, newly depositedeggs contained differently developed embryos resulting in anincubation period varying from 1 to 34 days. Each snail on averagelaid 6.0 eggs (range 1–16 eggs) during its life. Egg productionrate and hatching success in individuals kept in isolation werethe same as in individuals kept in pairs. Hatchlings measured0.49 mm in shell breadth. Snails grew throughout their life,but the growth rate declined significantly at a shell breadthof 1.3–1.4 mm, the point at which the onset of sexualmaturity occurred. Comparison of reproductive characters ofsnails living in leaf litter revealed that very small-sizedspecies produce few, but in relation to their shell size largeeggs, which they oviposit singly. ^*Present address: Zoological Institute, University of Basel,Rhein-sprung 9. CH-4051 Basel, Switzertand (Received 23 May 1988; accepted 11 July 1988)     

Internal embryonic brooding and development in the southern Australian micro-snail Tricolia rosea (Angas, 1867) (Vetigastropoda: Phasianellidae: Tricoliinae)*

ABSTRACT

Internal embryonic brooding has been suggested as an adaptation to enhance reproductive success in minute gastropods. It is rare in vetigastropods, previously known in only two species of Spectamen Iredale, 1924 (Solariellidae) from South Africa. Herein it is confirmed in the temperate Australian micro-snail Tricolia rosea (Angas, 1867), with up to 46 embryos observed within a gravid female. Embryos are brooded to an advanced stage, possessing a translucent, calcified shell and a calcified operculum. The initial protoconch is colourless, spirally sculptured and delineated by a consistent axial demarcation at the 0.75 whorl mark, when it measures 320?µm across. Beyond this, the second part of the protoconch is tinted pink, the strong spiral sculpture continues but the ribs and interstices are broader, with smoother surface microsculpture. At the 1.1 to 1.125 whorl mark the protoconch measures 400?µm across when transition to smooth teleoconch sculpture occurs. No brooded embryos possessed teleoconch sculpture. The potential relationship of protoconch morphology to embryonic development, hatching, feeding and release are considered. The mechanism of fertilisation is unknown, but embryos in a brood are at the same developmental stage. Unanswered questions in embryonic development and problems with protoconch terminologies in vetigastropods are also discussed.     

Effects of Diazocyclopentadiene (DACP) and Silver Thiosulphate (STS) on Ethylene Regulated Abscission of Sweet Pea Flowers (Lathyrus odoratus L.)

The ethylene production rate of cut sweet pea flower buds increased37-fold during the first 48 h of their vase life. This increasein ethylene production was accompanied by petal wilting at 72h and abscission of the buds 24 h later. Exposure of the cutspikes to the ethylene action inhibitor diazocyclopentadiene(DACP, 170 µI 1^-1) for 18 h under fluorescent lights delayedsubsequent wilting and abscission and promoted bud opening.Silver thiosulphate (0·2 mM) was more effective thanDACP, delaying wilting for longer and preventing abscissionentirely.Copyright 1995, 1999 Academic Press Ethylene, abscission, silver thiosulphate, diazocyclopentadiene, flower senescence, wilting, sweet pea, Lathyrus odoratus L     

Temporal effect of human oviductal cell and its derived embryotrophic factors on mouse embryo development.

Mouse embryos at different stages of development were cocultured with human oviduct cells or cultured in the presence of oviduct-derived embryotrophic factor-1, -2, and -3 (ETF-1, -2, and -3) for various amounts of time within the preimplantation period. Cocultures that included the period from 48 to 72 h post-hCG stimulated cell division and increased the cell numbers in the inner cell mass (ICM) of the exposed blastocyst. Exposure of embryos to oviductal cells from 96 to 120 h post-hCG increased the cell number in the trophectoderm (TE), blastocyst size, hatching rate, attachment, and in vitro spreading of the blastocyst. ETF-1 and ETF-2 affected embryos between 48 and 72 h post-hCG by increasing the number of cells in the ICM. In contrast, ETF-3 had a more profound effect on embryos that were exposed from 96 to 120 h post-hCG, where it mostly affected the development of TE cells, leading to higher hatching rate. Human oviductal cells improved mouse embryo development partly by the production of high molecular weight embryotrophic factors. These factors had differential effects on mouse embryo development.     

Myo-inositol Biosynthesis and Galactose Utilization by Wild Carrot (Daucus carota L. var. carota) Suspension Cultures

Wild carrot (Daucus carota var. carota) cell suspensions (63–120µm in diameter) were grown on a mineral salt medium containingdifferent carbon sources in the presence (10 mM) and absenceof myo-inositol. The data obtained after 14 and 21 days of growthshow that an external supply of myo-inositol is not essentialfor growth and development of wild carrot embryos. A linearrelationship was found between growth (d. wt) and embryo numberin the presence and absence of myo-inositol. Standard stock cell suspensions never exposed to exogenous myo-inositoland grown in the absence of 2, 4-D with glucose or galactoseas the carbon source synthesized radioactive myo-inositol whenexposed to D-[1–¹⁴C]glucose or D-[1–¹⁴C]galactose.Gas chromatographic analyses revealed the presence of myo-inositolin the bulk tissue grown in the presence of 2.25 µM 2,4-D with glucose, galactose, fructose or mannose as the solecarbohydrate. We could not detect any component indicating anisomer or a methylated derivative of an inositol in the tissueextracts. Stock cultures were maintained (with 2, 4-D) successfully forat least three successive sub-cultures on D-galactose as thesole carbohydrate. The growth achieved over this culture periodshowed that wild carrot cells used by us could quickly adaptto grow on D-galactose as rapidly as they grow on sucrose. Daucus carota L., wild carrot, suspension cultures, myo-inositol, galactose     

Embryonic, early larval development time, hatching mechanism and interbrood period of the sac-spawning euphausiid Nyctiphanes simplex Hansen

Females of the sac-spawning euphausiid Nyctiphanes simplex Hansenwere incubated under shipboard laboratory conditions to observethe embryonic and larval development time and hatching mechanism.Females ready to spawn have a pale pink ovary that extends fromthe back of the stomach to the first abdominal segment, fillingmost of the haemocoel. This species usually behaves as a totalspawner (produces one batch of oöcytes per cycle of theovary) leaving an ‘empty’ space in the cephalothoraxwhere the spent ovary is located. After spawning, the youngoöcytes mature and turn pale pink. The eggs do not havea measurable perivitelline space (PVS) in any of the embryonicstages (6.6 x magnification). The embryos hatch as nauplius(80–91 h after spawning, 16°C ± 1°C). Theyfurther develop into pseudometanauplii (PMN, 90–105 hafter spawning) and metanauplii (MN, 92–140 h after spawning)inside the ovigerous sac. The nauplius breaks the thin and fragilechorion by increasing the volume of the body and by using thefirst and second antennae. We call this an ‘expansion’hatching mechanism, the fifth distinct hatching mechanism observedso far among euphausiids. N. simplex larvae escape from theovigerous sac late in the MN stage (5 days after spawning),just a few hours before molting into calyptopis 1 (C1) (0.5–4h). This delayed release extends protection by the female, likelydecreasing the risk of predation or early cannibalism. Additionally,this may save energy by not swimming independently increasingthe time of not return if the calyptopis does not find favorablefeeding conditions. Females are not ready to spawn again untilat least two days after the previous batch of embryos leavesthe ovigerous sac. The interbrood period (IBP) observed rangedbetween 7 and 15 days at 16–18°C. This IBP is aboutone-fourth to half than was previously assumed for this speciessuggesting a significant underestimation of the fecundity ofthis species. N. simplex hatching success usually was 100%,except for a few females with all of their embryos dying duringembryonic development. Other females either molted before releasingthe embryos, or the oöcytes were spawned unfertilized (0%hatching success), particularly during winter conditions. Efficienthatching and late free-swimming strategy may partially explainwhy this species is the most abundant neritic euphausiid inthe southern part of the California Current System (CCS) andin the Gulf of California.     

BROODING AND NON-BROODING DACRYDIUM (BIVALVIA: MYTILIDAE): A REVIEW OF THE ATLANTIC SPECIES

The Atlantic species of the marine bivalve genus Dacrydium arereviewed, with particular emphasis on their hinge and protoconchcharacters. The basic groundplan of a Dacrydium comprises afunctional primary ligament, a paired series of primary teeth,and a posterior row of secondary teeth separated from the latterby a secondary ligament; this can be transformed into a singleseries either by loss of the secondary ligament and mergingprimary and secondary teeth, or by loss of secondary teeth andligament through paedomorphosis. Twelve species are recognized, of which eleven are illustrated.One abyssal species is not separable morphologically from theIndian Ocean D. speculum Poutiers, 1989 and is new to the Atlantic;four new species (D. wareni, D. dauvini, D. filiferum and D.balgimi) are described; a Caribbean form which is hardly distinctfrom the Eastern Pacific D. elegantu-lum Soot-Ryen, 1955, isdescribed as a new subspecies D. e. hendersoni. The larvae are brooded in D. hyalinum (Mon-terosato, 1875),D. viviparum Ockelmann, 1983 and D. balgimi. The brooding specieshave larger larvae (protoconch 210 to 315 µm long) thanthe non-brooding (protoconch 120 to 150 µm long), andreach a smaller adult size (1.4 to 3 mm instead of 4.5–5mm). A phylogenetic reconstruction is attempted using parsimonyanalysis of hinge and shell characters as well as the brooding/nonbrooding character. (Received 22 October 1996; accepted 28 November 1996)     

Expression of the gus A gene in embryogenic cell lines of Sitka spruce following Agrobacterium-mediated transformation

Transformation of Picea sitchensis (Bong) Carr. was investigatedby incubating embryogenic cell lines, initiated from immatureand mature zygotic embryos, with a supervirulent strain of Agrobacteriumtumefaciens. The latter carried a gus A-intron gene. Transientgene expression was determined histochemically by recordingthe number of distinct areas of ß-glucuronidase (GUS)activity. Maximum expression of the gus gene was achieved witha bacterial suspension with an OD₆₀₀ of 0.8–1.1 dilutedwith an equal volume of MPM medium, Inoculation of cells withbacteria for 30 mm, 72 h co-cultivation period and exposureof Agrobacterium and plant cells to 50 µM acetosyrmngone.These results are discussed in relation to Agrobacterium-mediatedgene delivery for the stable transformation of Sitka spruceand other conifers. Key words: Sitka spruce, Agrobacterium, transformation, embryonal suspensor masses, GUS activity     

Feeding and growth rates of the doliolid, Dolioletta gegenbauri Uljanin (Tunicata, Thaliacea)

The goal of this research is to enhance our knowledge of thecontributions of doliolids to the planktonic community as consumersand secondary producers. The objectives are to quantify feedingand growth rates of Dolioletta gegenbauri gonozooids at fourfood concentrations and four temperatures in order to determinetheir impact as grazers throughout the water column. Althoughdoliolids are abundant in numerous regions of the coastal ocean,and are considered to be major planktonic grazers, data on ratesof feeding and growth are scarce. Laboratory experiments wereconducted at 16.5, 20, 23.5 and 26.5°C to quantify removalof a 50:50 volumetric concentration of Thalassiosira weissflogiiand Rhodomonas sp. at four different food concentrations of20, 60, 160 and 390 µg C l^–1. Results from theseexperiments suggest that clearance rates are similar at concentrationsfrom 20 to 60 µg C l ^–1, and decrease as the foodconcentrations increase to 160 and 390 µg C l ^–1.The ingestion rates increase over a range of phytoplankton concentrationsfrom 20 to 160 µg C l ^–1, then decrease when abnormallyhigh concentrations of 390 µg C l ^–1 are offered.Clearance and ingestion rates increase as temperature increasesfrom 16.5 to 26.5°C. The exponential growth rates rangefrom k = 0.2–0.7, with the lowest rates occurring at thehighest food concentration. Growth rates increase with increasingtemperature from K = 0.1–0.3 at 16.5°C to 0.45–0.7at 26.5°C. In each case, the small- and medium-sized zooidshad higher growth rates than the larger gonozooids. These resultssuggest that doliolid feeding and growth rates are a functionof environmental food concentrations and temperatures, and implythat they can be important consumers in a changing neritic environment.     

Application of synchrotron-radiation-based computer microtomography (SRμCT) to selected biominerals: embryonic snails, statoliths of medusae, and human teeth

Synchrotron-radiation-based computer microtomography (SRμCT) was applied to three biomineralised objects First, embryonic snails of the freshwater snail Biomphalaria glabrata, second, rhopalia (complex sense organs) of the medusa Aurelia aurita, and third, human teeth. The high absorption contrast between the soft tissue and mineralised tissues, i.e. the shell in the first case (consisting of calcium carbonate) and the statoliths in the second case (consisting of calcium sulphate hemihydrate), makes this method ideal for the study of biomineralised tissues. The objects can be non-destructively studied on a micrometre scale, and quantitative parameters like the thickness of a forming a snail shell or statolith crystal sizes can be obtained on a length scale of 1–2 μm. Using SRμCT, the dentin–enamel border can be clearly identified in X-ray dense teeth.     

Cyst formation: an important mechanism for the termination of Scrippsiella trochoidea (Dinophyceae) bloom

A sediment trap study was conducted at Daya Bay, South ChinaSea, to investigate the relationships between encystment andpopulation dynamics of Scrippsiella trochoidea from December1999 to January 2001. A dense bloom of S. trochoidea occurredduring the study period from August to September 2000, withthe maximum cell number of 3.18 x 10⁴ cells mL^–1.Two morphotypes of cysts, one with a thick calcareous wall (calcifiedcyst) and another without the obvious calcareous cover (non-calcifiedcyst), were observed during this investigation. The morphologicaland excystment characteristics of these two cyst types werestudied as well. Mass encystments of S. trochoidea, with themaximum of 3.05 x 10⁵ cysts m^–2 d^–1for calcified cyst, and 1.54 x 10⁷ cysts m^–2 d^–1for non-calcified cyst, coincided with the maximum abundanceof the vegetative cells. Encystment caused the transfer of atotal of 2.24–4.49 x 10⁸ cells m^–2 vegetativecells from the water column to the sea bottom during the bloomand resulted in a considerable loss of the bloom population.High assemblages of cysts of S. trochoidea were detected inthe surface sediments as well. This rich ‘seed bed’in the surface sediments caused by the high efficiency of encystmentafter blooms acting as a benthic reservoir for future vegetativepopulation, together with the short dormant period (15–26days) and high germination rate (50–90%), may explainthe repeated occurrence of S. trochoidea blooms in Daya Bay.     

THE RATIO OF CALCAREOUS AND ORGANIC SHELL COMPONENTS OF FRESHWATER SPHAERIID CLAMS IN RELATION TO WATER HARDNESS AND TROPHIC CONDITIONS

Shells from 14 populations of sphaeriid clams including Sphaeriumstriatinum, S. simile, Pisidium walkeri, Musculim partumeiumand M. iransversum were analyzed for organic carbon (µgCmg^–1 shell), nitrogen (µg,N mg^–1 shell) andCaCO_s (%CaCO₃ of total clam dry weight). Habitat waters wereassessed for total hardness (expressed as ppm CaCo₃), ppm Ca,ppm Mg, conductivity (µmho) and suspended organic Carbon(µgCl^–1). For all populations, shell organic C andN are positively correlated and there is an inverse relationshipbetween the amounts of shell CaCO₃ and shell organic carbon.Trophic considerations give the best correlation with shelltype at the generic level of consideration since species ofMusculium are found at the opposite end of the trophic scale(eutrophic) from all other populations studied. For S. striatinum,the most extensively studied species, the amount of shell CaCO₃is inversely related to water hardness. The selection of shellsin the Sphaeriidae is discussed in relation to structural-functionalneeds and habitat conditions ¹ Present Address: Department of Biology, Syracuse University,Syracuse, New York 13210, U.S.A. ² Present Address: Department of Zoology, Miami University,Oxford, Ohio 45056, U.S.A. (Received 5 December 1978;