A study of the morphology,ultrastructure, and histochemistry of the food of the pediveliger of Ostrea edulis

During the settlement of Ostrea edulis L., the foot of the pediveliger implements temporary attachment and cementing. The morphology of the foot has been re-examined to try to clarify its role during this period. The foot has a highly developed nervous system and musculature, but mainly consists of sub-epidermal gland cells of 9 different types. Cells of the first gland contain acid mucopolysaccharide, and open over the ventral surface of the foot. The second type of gland contains a neutral glycoprotein, and opens at the tip of the foot. The third gland opens mid-ventrally half way down the foot, and largely contains an aromatic protein. The fourth type of gland contains mainly proteoglycan, and the cells open mid-ventrally behind the third gland. The fifth gland contains acid mucopolysaccharide, and the cells open onto the heel of the foot. The sixth, seventh, eighth and ninth glands contain a variety of acid mucopolysaccharides, proteoglycans and possibly a neutral glycoprotein; their cells open into the byssus duct, which discharges at the base of the heel. The grouping of the glands, position of the openings of the cells, and histochemical properties of the secretions suggest that they may be involved in localised and general adhesion of the foot during temporary attachment, as well as in cementing of the larva.     

Attachment of the larva of the ascidian Diplosoma listerianum

Histological and ultrastructural studies of free-swimming and attached larvae of Diplosoma listerianum Milne Edwards indicate that initial attachment is by means of papillae, which secrete an adhesive substance. The source of the adhesive is the central mass cells of the papilla. These cells secrete large electron-dense granules, and also a mass of reticular material. The granules give rise to the adhesive seen in sections and scanning electron micrographs of attached larvae, although the reticular material may also contribute to the adhesive. Histochemical tests show that protein is present in the granules, together with a small amount of carbohydrate. The reticular material contains sulphated acid mucopolysaccharide.     

Origin and nature of spermatophoric mass of the spiny lobster Panulirus homarus

In the spiny lobster Panulirus homarus (Linnaeus) the spermatophoric mass consists of a highly convoluted tube, containing the sperm mass and a gelatinous matrix. The spermatophoric components have their origin in the proximal vas deferens and the completed spermatophoric mass is stored within the distal dilated part of the vas deferens. In the proximal vas deferens the inner glandular epithelial cells give rise to leaf-like typhlosoles which gradually diminish in size and finally disappear in the distal vas deferens. The secretions originating from the typhlosole form the outer gelatinous matrix of the spermatophoric mass. Histochemical observations reveal that the wall of the spermatophore tube consists of neutral mucopolysaccharide, whereas the sperm mass and the gelatinous matrix are rich in acidic mucopolysaccharides. Both polyacrylamide gel electrophoresis and agarose gel electrophoresis were used to separate the acidic mucopolysaccharide (AMPS) complexed with the proteins and the simple proteins. In addition, AMPS have been isolated following the method of Rahemtulla and Lovtrup (1974) and used for electrophoresis. In both the polyacrylamide gel electrophoresis and agarose gel electrophoresis, the extracted acid mucopolysaccharides give only a single fraction which, in comparison with the standard acid mucopolysaccharides, corresponds to chondroitin sulphate. The functional significance of the chondroitin sulphate and neutral mucopolysaccharides in the spermatophoric components of P. homarus is discussed in relation to their functional role in spermatophore hardening and protection of the delicate spermatozoa during their prolonged storage on the sternum of the female.     

Uptake of bacterial extracellular polymer by a deposit-feeding holothurian (Isostichopus badionotus)

A laboratory experiment shows that exopolymer (=mucopolysaccharide) produced by the estuarine marine bacterium Pseudomonas atlantica can be a source of nutrition for the deposit-feeding holothurian Isostichopus badionotus (Selenka). Holothurians fed sediment prepared with ¹⁴C-labeled exopolymer incorporated radioactive carbon into their rete mirabile.     

Inhibition of attachment of larval barnacles,Balanus amphitrite,by bacterial surface films

Films of bacteria on solid substrata can positively or negatively influence the attachment of marine invertebrate larvae. Effects of marine bacteria on the attachment of cypris larvae of the barnacle Balanus amphitrite Darwin were examined in the laboratory. Bacteria, grown to mid-exponential phase and allowed to adsorb irreversibly to polystyrene petri dishes, attached in densities of 10⁷ cells cm^-2. Assays (22h) were used to compare the effects of adsorbed cells of 18 different bacterial species on larval barnacle attachment. Most of the adsorbed bacteria either inhibited or had no effect on larval attachment compared to clean surfaces. Experiments testing the effect of larval age on barnacle attachment were conducted with six species of bacteria and showed that older larvae attached in higher percentages to clean surfaces and that bacterial films generally inhibited larval attaschment. Both the species of bacteria and the in situ age of the adsorbed bacteria affected barnacle attachment response: older films of Deleya (Pseudomonas) marina were more inhibitory. Bacterial extracellular materials may be involved in the inhibitory process.     

The role of mucus in particle processing by suspension-feeding marine bivalves: unifying principles

Contemporary research on bivalve suspension-feeding has revealed a diversity of particle processing mechanisms depending on the anatomy and functioning of the pallial organs involved. On the biochemical level, however, some evidence of homogeneity has emerged concerning the types of mucopolysaccharide associated with particle processing. The present study uses both previous data and original research combining video endoscopy and mucocyte mapping to further explore the relationships between pallial organ topography, functional correlates, direction of current flow, and mucocyte secretion type. Five species representing five different families and all four major gill types are represented: Mytilus edulis, Placopecten magellanicus, Crassostrea virginica, Mya arenaria, and Spisula solidissima. Viscous acid or acid-dominant mucopolysaccharides are used when particle transport occurs on an exposed surface, or on a structure leading directly to such a surface, counter to the prevailing current flow. Associated functions are indiscriminate transport in gill ventral particle grooves and rejection of pseudofeces. Lower-viscosity mixed mucopolysaccharides are used when particle transport is on an enclosed or semi-enclosed surface, leading to other such surfaces, and with the current flow. Associated functions are transport of particles destined for ingestion, and ingestion itself. Low-viscosity neutral mucopolysaccharides are found in regions where reduction of mucus viscosity is important, such as the areas of the labial palps responsible for fluidization of the high-viscosity mucus-particle cord of the gill ventral particle groove prior to particle extraction. There thus appears to be a specialization of mucus type corresponding to functional specialization of the various pallial organs in suspension-feeding marine bivalves. Received: 10 October 1996 / Accepted: 7 May 1997     

Whitefly wax as a cue for phoresy in the broad mite, <Emphasis Type="Italic">Polyphagotarsonemus latus</Emphasis>(Acari: Tarsonemidae)

Summary. Broad mite, Polyphagotarsonemus latus (Acari: Tarsonemidae) exhibits a specific phoretic relationship with whiteflies. Under field conditions most broad mites, caught in sticky traps, are attached to whiteflies. Under laboratory conditions, attachment occurs equally well in the dark and light. Mites do not differentiate between the sexes of their phoretic host Bemisia tabaci. However, mite attachment to B. tabaci is greatly diminished by washing the host with various organic solvents, chloroform in particular. The effect of whitefly waxy particles on broad mite behavior was studied using wax from the whitefly Aleyrodes singularis and from the mealybug Planococcus citri. Broad mites were not only attracted specifically to the A. singularis waxy particles-treated leaf areas but were also attached to leaf trichomes in this area. The results of this study suggests the importance of olfactory cues from the whitefly waxy particles in the recognition process of the phoretic host and/or the induction of the attachment behavior to whitefly legs or leaf trichomes.     

Variation in strength of attachment to the substrate explains differential mortality in hybrid mussel (Mytilus galloprovincialis and M. edulis) populations

Mussel samples were collected from a hybrid mussel (Mytilus galloprovincialis and M. edulis) Population at Croyde, southwest England, in January, March and May 1990. The strength of attachment of each mussel to the substrate was measured with a spring balance. A number of diagnostic characters were also recorded. These are shell lengh, width and height, mantle colour and genotype at two allozyme loci, esterase-D and octopine dehydrogenase. Multiple-regression analysis was used to assess the effect of the diagnostic characters on strength of attachment as dependent variable. Mussels possessing the relatively high shells and darker mantle colouration characteristic of M. galloprovincialis had higher values, on average, for strength of attachment than mussels resembling M. edulis. Phenotypically intermediate mussels had intermediate values for strength of attachment. The results suggest an adaptive difference which can account for reports of differential mortality acting in favour of M. galloprovincialiis.     

Glucose and glycerol uptake by isolated zooxanthellae fromCassiopea xamachana: Transport mechanisms and regulation by host homogenate fractions

We investigated the mechanisms by which glycerol and glucose enter freshly isolated zooxanthellae (FIZ),Symbiodinium microadriaticum Freudenthal, of the mangrove jellyfish,Cassiopea xamachana Bigelow, and the specific sites of host factor interaction. Glucose entry into FIZ is accomplished by a Na⁺-dependent symport system driven by an electrochemical gradient generated via a Na⁺-K⁺ ATPase. Inhibition of glucose uptake by a low molecular weight fraction of host homogenates [mol. wt<2 kilodaltons (kD)] occurs through the interaction of a putative host factor with the carrier protein and not the ATPase. Glycerol entry is apparently accomplished by simple or facilitated diffusion and is not affected by host homogenate fractions.     

Hydrodynamic stress and habitat partitioning between indigenous (Perna perna) and invasive (Mytilus galloprovincialis) mussels: constraints of an evolutionary strategy

The ability of a mussel to withstand wave-generated hydrodynamic stress depends mainly on its byssal attachment strength. This study investigated causes and consequences of different attachment strengths of the two dominant mussels species on the South African south coast, the invasive Mytilus galloprovincialis and the indigenous Perna perna, which dominate the upper and the lower areas of the lower balanoid zone, respectively and co-exist in the middle area. Attachment strength of P. perna was significantly higher than that of M. galloprovincialis. Likewise solitary mussels were more strongly attached than mussels living within mussel beds (bed mussels), and in both cases this can be explained by more and thicker byssal threads. Having a wider shell, M. galloprovincialis is also subjected to higher hydrodynamic loads than P. perna. Attachment strength of both species increased from higher to lower shore, in response to a gradient of stronger wave action. The morphological features of the invasive species and its higher mortality rates during winter storms help to explain the exclusion of M. galloprovincialis from the low shore. The results are discussed in the context of the evolutionary strategy of the alien mussel, which directs most of its energy to fast growth and high reproductive output, apparently at the cost of reduced attachment strength. This raises the prediction that its invasive impact will be more pronounced at sites subject to strong but not extreme wave action.     

Tube-building by a marine meiobenthic harpacticoid copepod

Pseudostenhelia wellsi Coull and Fleeger is a meiobenthic harpacticoid copepod inhabiting muddy, estuarine sediments. All individuals observed, and all ages including nauplii, build and inhabit elongate, mucous tubes which may extend to a depth of 3.9 mm into the sediment. Many of the narrow tubes, 0.27 mm in diameter, have a tube-cap extending 0.32 mm above the sediment-water interface. The tubes are a matrix of fine silt, sand and detritus cemented with a mucopolysaccharide, as shown by the Periodic Acid Schiff stain, apparently secreted from glands in the ventrolateral margin of the cephalothorax. Upon addition to sterile, homogeneous sediment, P. wellsi quickly (within 1 to 2 h) transform the upper 0.4 cm to a cohesive conglomerate of tubes, silt-clay particles and mucous filaments.     

Benzoic acid, a stimulant of odorant receptors of Bombyx mori, is rapidly metabolized to N-benzoylserine on the antennae

Summary. Benzoic acid is a known stimulant of olfactory receptor cells in trichoid sensilla on the antennae of adult females of Bombyx mori. Exposure of freshly excised antennae, from B. mori females and males, to vapours of [¹⁴C]-benzoic acid revealed that the adsorbed acid is rapidly metabolized (15% conversion in 10 s), suggesting the presence of a highly active enzymatic system. The major product could be identified as N-benzoylserine by use of [²H₅]-benzoic acid, and mass spectrometry linked to a gas chromatograph (GC-MS) or a liquid chromatograph (LC-MS), for analysis of the metabolites. The conjugation of benzoic acid with an amino acid apparently represents a novel mode for the rapid deactivation of odorant molecules. Received 26 March 2001.     

Extraction,isolation, and identification of various environment friendly components from cock's comb (Celosia argentea) leaves for allelopathic potential

In this study, the aqueous extract of leaves of the plant cock's comb (Celosia argentea) was fractioned into five batches using n-hexane, dichloromethane, ethyl acetate, and n-butanol, respectively. High performance liquid chromatography analysis of the methanol re-dissolved residues of 3 polar fractions, including dichloromethane, ethyl acetate, and n-butanol separately which exhibited altogether 12 compounds identified as 4-hydroxycinnamic acid (p-coumaric acid), 4-hydroxybenzoic acid, protocatechuic acid, caffeic acid, m-coumaric acid, gallic acid, 4-hydroxybenzaldehyde, m-hydroxybenzaldehyde, 2,4-dihydroxybenzoic acid, pyrogallol, 3,5-dihydroxybenzoic acid, and genetic acid was categorized by retention time using known compounds. Similarly, the mass confirmation of most of the identified compounds was achieved by injecting 20 µl of composite sample of these three polar fractions into liquid chromatography–mass spectrometry. Similarly, the physiochemical analysis of C. argentea aqueous extract showed the presence of various constituents such as phenols, flavonoids, tannins, and glycosides. All four organic fractions were examined for phytotoxicity against Lepidium sativum. Analysis of variance showed significant differences among these fractions against germination and growth of L. sativum. Further allelopathic effects appeared to be concentration dependent. Thus, C. argentea may prove effective as a substitute of herbicides to control the weeds like L. sativum from attacking crops and to protect the environment from the health hazard effects of herbicides.     

Experimental analysis of byssus thread production by Mytilus edulis and Modiolus modiolus in sediments

Laboratory experiments have been conducted on byssus thread production by two species of mussel, Mytilus edulis L. and Modiolus modiolus (L.), representing the epibyssate and endobyssate species of the Mytilidae, respectively. Mussels were placed in seven particle sizes of sediment ranging from 50 m to 16 mm for 12 d. The number of byssus threads per mussel, length of threads, number of threads per particle and size of pads were then recorded. Modiolus modiolus (endobyssate) produced more threads than Mytilus edulis (epibyssate). M. edulis produced most threads in the size range 2 to 16 mm and Modiolus modiolus in the size range 500 to 1000 m. M. modiolus produced longer threads than Mytilus edulis. Both species produced longer threads in particle sizes finer than 2 mm. M. edulis produced smaller pads than Modiolus modiolus. Both species produced larger pads in sediments coarser than 1 000 m. Results were also analysed in terms of attachment units (a thread attached to one or more particles, or a particle to which more than one thread is attached). The thread:particle ratio of the attachment units ranged from ca. 1:10 in the finest sediments to ca. 10:1 in the coarsest sediments. Mytilus edulis attached more threads to single particles (72% of attachment units) than Modiolus modiolus (37% of attachment units). M. modiolus had a wider spread of ratios in the finer particle sizes than Mytilus edulis. The significance of our results are discussed in relation to the ecology and palaeoecology of epibyssate and endobysste species.     

Relationship between duration of cyprid attachment and drag forces associated with detachment ofBalanus amphitrite cyprids

Relationships between the duration of antennular attachment of cyprids ofBalanus amphitrite and forces associated with detachment were assessed in a small flume. For six of twelve sets of measurements obtained on separate dates between 27 January and 2 June 1989, drag forces associated with detachment were significantly and positively correlated with duration of attachment within the range 10⁰ to 10³ s. In general, the instantaneous drag force exerted on cyprids at detachment was more strongly correlated with duration of attachment than was either the maximum or mean force exerted prior to detachment. Our indirect method of estimating drag forces from shear velocities measured 0.5 cm from cyprids (i.e., 10 body lengths) probably underestimated the true association between detachment force and attachment time since the spatial coherence of shear velocities characterizing turbulent flows in the flume was not strong at these scales. The relationship between attachment time and drag force at detachment suggests that spatial and temporal variability in fluid forces in situ may contribute to stochastic variations in intensities of settlement ofB. amphitrite. Stresses sustained by the antennular adhesive of cyprids ofB. amphitrite were calculated to standardize forces to the area of antennular contact and permit comparisons with similar measurements made onSemibalanus balanoides by other investigators. Stresses sustained by the adhesive ofB. amphitrite were an order of magnitude lower than those reported for the adhesive of the larger cyprids ofS. balanoides. This difference may reflect differences in the stress sustainable by the adhesive secreted onto antennular pads when loaded purely in tension (measured previously) rather than in a combination of shear and tension (calculated here). Alternatively, there may be interspecific differences in the adhesive used for reversible attachment or in behavioral responses of exploring cyprids to strong flow.     

The wartbiter spermatophore and its effect on female reproductive output (Orthoptera: Tettigoniidae,Decticus verrucivorus)

Summary Male wartbiters Decticus verrucivorus transfer elaborate spermatophores to females during copulation. The spermatophore is attached externally to the female's genitalia and consists of two parts: a large, gelatinous, sperm-free portion, the spermatophylax, eaten by the female after mating; and a sperm-containing ampulla, eaten after the spermatophylax has been consumed. Since females take longer to eat larger spermatophylaxes, the duration of ampulla attachment is positively correlated with spermatophylax size. Two series of experiments were carried out, one in which the size of the spermatophylax consumed by females and the duration of ampulla attachment were manipulated in concert and another in which they were manipulated independently. Some females were also maintained on a protein-free diet and either supplied with or deprived of spermatophylax material. The amount of protein in the diet, but not the amount of spermatophylax material consumed, influenced female longevity, lifetime fecundity, and egg weight. When females were deprived of the spermatophylax, an experimental increase in the duration of ampulla attachment induced longer periods of unreceptivity in females after mating, a more rapid onset of oviposition, and an increased oviposition rate. Consequently, the number of eggs laid by females during their nonreceptive refractory periods increased significantly with increasing duration of ampulla attachment up to 180 min; however, there was no significant increase in the number of eggs laid beyond 180 min of ampulla attachment. This closely corresponds to an ampulla attachment duration of 188 min expected when a male transfers a natural spermatophylax of mean size to the female. These results suggest that the amount of ejaculate transferred to the female, and not the amount of spermatophylax material per se, is the factor controlling female receptivity and oviposition behavior. Since we did not detect any effect of spermatophylax consumption on female fecundity or egg weight, we conclude that the spermatophylax of D. verrucivorus functions primarily as a sperm protection device rather than as a form of male parental investment.     

Development and environmental application of a genus-specific quantitative PCR approach for Pseudo-nitzschia species

Quantitative polymerase chain reaction (qPCR) for the identification and quantification of microbes has become a common tool for the study of harmful algal blooms (HABs). We developed a qPCR method for the diatom genus Pseudo-nitzschia. Several species of this genus form toxic blooms through the production of the neurotoxin domoic acid (DA). Outbreaks of toxicity attributed to DA along the US west coast have caused sickness and death of marine mammals and seabirds through food web contamination. The method developed here quantifies Pseudo-nitzschia spp. at low abundances in natural samples, thereby, providing a method to improve our understanding of the environmental conditions leading to blooms of these species. This has been accomplished previously by techniques for identification and quantification that are slow and laborious compared to qPCR. The approach was successfully tested and validated using eight species of Pseudo-nitzschia and 33 non-target organisms and employed to follow local bloom dynamics.     

Reproduction,growth and mortality of the holothurian Cucumaria pseudocurata

Certain phases of the ecology of a sea cucumber (Cucumaria pseudocurata Deichmann, 1938) population at Shell Beach, Sonoma County, California (USA), were investigated. The sea cucumber is a brooder, lays large (1 mm diameter) yolky eggs, which undergo direct development. It forms large aggregations in the rocky intertidal zone, extending from the lower boundary of the Mytilus californianus beds to about zero tide-level. The gonad cycle was followed from September, 1970 to January, 1972. Spawning occurred in January, 1971 and January, 1972. The holothurians produce from 1 to as many as 340 eggs, depending on the size of the individual; they completely spawn-out in a few days. Hatching takes place about 1 month after egg laying. The main sources of mortality are predation by the sea star Pycnopodia helianthoides and wave action. Survival of the young is negatively correlated with density of the aggregation. C. pseudocurata increases in weight 10 to 30 fold in the first year, but much more slowly in later years. Individuals live perhaps 5 years or more, and do not become reproductive until about the third year. Dispersal is accomplished by the young being washed away to a suitable new habitat. The upper range of the intertidal distribution is determined by exposure to air, and the lower range by P. helianthoides predation.Contribution No. 1 of the Bodega Marine Sciences Association.     

Abundant ketone isolated from oily Plakortis sponge demonstrates antifouling properties

Marine sponges of the Genus Plakortis are typically unfouled; they can have a distinctive pleasant smell and an oily surface. A significant quantity of fragrant oil was obtained from a Jamaican Plakortis sp. by cryo-trap. The oil was determined to be exclusively 2-decanone. The antifouling character of the oil was evaluated by its effects on surface attachment of a gram negative bacterial model using confocal fluorescence microscopy as well as its effects on the attachment of Dreissena polymorpha (zebra mussel). The ketone dramatically inhibited attachment of the bacteria and zebra mussels. The results suggest that the oil impacts establishment of related epifauna on the Plakortis sponge in nature. Although the aliphatic ketone alone is not a potential commercial alternative for antifouling coatings, incorporating the functionality into coating design should be considered in future studies.     

Characterizing the resident, fermentative microbial consortium in the hindgut of the temperate-zone herbivorous fish, Hermosilla azurea (Teleostei: Kyphosidae)

The zebraperch, Hermosilla azurea Jenkins and Evermann, a warm-temperate marine fish species with a strictly macroalgal diet, has a relatively long digestive tract with an enlarged hindgut and an associated blind caecum (HC). In zebraperch sampled off Santa Catalina Island, California (33°19′42′′N; 118°18′37′′W) in years 1995 through 2001, direct cell counts, gut epithelium assessment of bacterial attachment, and short-chain fatty acid (SCFA) analyses verified that the zebraperch HC possesses a dense and morphologically diverse, fermentative microbiota. Bacterial cell counts and morphological diversity were significantly higher in HC contents compared to anterior gut regions, suggesting that microbial populations were growing along the digestive tract. Similarly, electron micrographs of the HC epithelium revealed attached microbes, further supporting the possibility that these organisms constitute resident microbiota. Five different SCFAs were detected in all three regions of the digestive tract, but levels were up to three times greater in HC contents. Acetate was consistently the prevailing SCFA in all gut regions. Sequence analysis of bacterial 16S rDNA was used to identify predominant bacterial groups in HC contents. Of the seven main bacterial types identified, Enterovibrio spp. were the dominant bacteria in HC contents followed by species of Bacteroides,Faecalibacterium, and Desulfovibrio. Taken together, our findings show that the zebraperch HC harbors a consortium of microbes that appears to assist in the breakdown of algal polysaccharides in the herbivorous diet of the fish.