Larval and post-larval morphogenesis in the gutless protobranch bivalve Solemya reidi (Cryptodonta: Solemyidae)

Structural changes occurring in the alimentary tract, perivisceral cavity, foot, hypobranchial gland, and gills of larval and post-larval Solemya reidi, a gutless protobranch bivalve, were examined using both light and electron microscopy at 1, 3, 5, 18, 34 and 42 d after fertilization. A fully developed mouth, esophagus, and anus, together with the rudiments of a stomach and rectum are present before metamorphosis. At metamorphosis, the cells making up the dorsal and lateral walls of the stomach dissociate, and by 18 d the mouth and anus are the only remaining portions of the alimentary tract. The larval test is ingested into the perivisceral cavity at metamorphosis and its autolysis continues through at least the 42nd day after fertilization. The foot, gills, and hypobranchial gland, poorly developed at metamorphosis, develop slowly and are still undergoing extensive morphogenesis at 42 d.Harbor Branch Oceanographic Institution Contribution No. 601.     

Translocation of fixed carbon from symbiotic bacteria to host tissues in the gutless bivalve Solemya reidi

Naturally-occurring lipophilic inducers of larval settlement and metamorphosis wer isolated and identified for Phragmatopoma californica, a gregarious tube worm from southern California. Organic solvent extraction of the sand/organic cement matrix of tubes diminished the inducing capacity of the tube matrix. The inducing capacity was restricted to a single, highly active, HPLC-purified fraction of the organic solvent extract. Chemical analysis of this fraction revealed a mixture of free fatty acids (FFAs), dominated by eicosapentaenoic acid (20:5, 20%), palmitic acid (16:0, 14%) and palmitoleic acid (16:1, 12%). In assays of the nine FFAs that each contributed 3% or more to the active fraction, only 16:1, 18:2, 20:4 and 20:5 induced larval settlement and metamorphosis, while the others were ineffective. The larval response was contact-dependent, highly specific, and concentration-dependent, with a significant response to 16:1 and 20:4 at as low as 10 g FFA spread onto 1 g of sand (surface area 36 cm²). Active FFAs were extracted at approximately 14 g g^-1 sand from the tube matrix, although the levels encountered by larvae in nature are believed to be higher.     

Development of the pericalymma larva of Solemya reidi (Bivalvia: Cryptodonta: Solemyidae) as revealed by light and electron microscopy

Solemya reidi Bernard 1980 is a gutless protobranch bivalve known to possess intracellular chemoautotrophic bacterial symbionts in its gill. A light and electron microscope study on the embryology and larval development of S. reidi provides data for the bivalve Subclass Cryptodonta. S. reidi spontaneously spawned large eggs (271 m in diameter), which developed within individual gelatious egg capsules. The first several cleavages were equal and a distinct molluscan cross was formed at the animal pole of the embryo, features previously unreported in bivalve development. Lecithotrophic pericalymma larvae (similar to the larvae of paleotaxodont protobranch bivalves and aplacophoran molluscs) hatched at 18 to 24 h and remained in the water column for a further 5 d at 10°C. At hatching, larvae measured from 360 to 440 m in length and from 225 to 265 m in cross-sectional diameter. Definitive adult structures developed within an epithelial locomotory test entirely covered with compound cilia. The test histolysed at metamorphosis and was ingested throught the mouth into the perivisceral cavity. Length and height of the shell following metamorphosis was 433 m (±42 m, n=16) and 282 m (± 29 m, n=13), respectively. Primary data and data from the literature show that the type of larval development in both paleotaxodont and cryptodont bivalves cannot be reliably estimated from egg or prodissoconch sizes.     

Oxygen transport by the hemocyanin of the protobranch mollusc Solemya reidi

Oxygen transport by the hemocyanin of the protobranch mollusc Solemya reidi Bernard was studied in native hemolymph samples. Clams were collected from two different reducing environments, beneath log booms in Alberni Inlet, British Columbia, Canada, and from the sewage effulent in Santa Monica Bay, California, USA. The hemocyanin concentration in a pooled hemolymph sample (n = 10 individuals) was 33.5 mg ml⁻¹. The mean hemolymph pH of five Alberni clams maintained for 3 wk in mud was 7.96 ± 0.06. No significant variation in hemocyanin oxygen-affinity or cooperativity was found for hemocyanin in whole hemolymph samples from these five individually studied clams. There was a significant difference only at 15 °C in the oxygen affinity of hemocyanin in pooled whole-hemolymph samples from S. reidi collected from Alberni Inlet compared with clams collected from Santa Monica Bay. Little effect of temperature on hemocyanin oxygen-affinity was found for temperatures below 20 °C; above 20 °C, the oxygen affinity was reduced for clams from both sites. Temperature and pH had no apparent affect on hemocyanin cooperativity. Moderate Bohr shifts were found at all temperatures examined. The presence of physiologically relevant concentrations of thiosulphate in hemocyanin samples resulted in a decrease in hemocyanin oxygen-affinity, opposite to the effect on hemocyanin found for the hydrothermal vent crab Byth-ograea thermydron, but thiosulphate had no effect on hemocyanin cooperativity. Received: 2 September 1997 / Accepted: 6 February 1998     

Association of bacteria with bryozoan larvae

Seven species of marine bryozoans were examined for the presence of bacteria associated with the larvae. In three species (Bugula neritina, B. pacifica, and B. simplex), rod-shaped bacteria were consistently observed in the pallial sinus of the larvae, independent of geographic location, year, or season. Larvae of closely related bryozoans (B. stolonifera, B. turrita, Scrupocellaria bertholetti, and Tricellaria occidentalis) occurring sympatrically with those containing bacteria lacked this apparent symbiosis. Those bryozoans examined that possessed the bacteria-larva association are among the most frequently encountered fouling bryozoans.     

Localization of RubisCO and sulfur in endosymbiotic bacteria of the gutless marine oligochaete Inanidrilus leukodermatus (Annelida)

 The chemoautotrophic potential of the two co-occurring larger and smaller bacterial endosymbionts of the gutless marine oligochaete Inanidrilus leukodermatus was determined using immunocytochemistry. An antibody directed against the Form I of ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO), the key CO₂-fixing enzyme of the Calvin–Benson cycle, consistently labeled the larger symbionts. Electron microscopic spectroscopy showed that the larger symbionts contained sulfur in intracellular globules and to a lesser degree in the cytoplasm. The presence of RubisCO and sulfur indicates that the larger endosymbionts of I. leukodermatus are chemoautotrophic sulfur-oxidizing bacteria. In contrast, no RubisCO or sulfur was detected in the smaller endosymbionts of this host. Received: 28 September 1999 / Accepted: 29 May 2000     

Acute and chronic toxicity of tributyltin (TBT) to pediveliger larvae of the bivalve Scrobicularia plana

A static renewal test was run for 30 d to assess the effects of the antifoulant tributyltin (TBT, nominal concentrations of 50, 125, 250 and 500 ng Sn l^-1) on the survival and growth of 240 m-long pediveliger larvae of the bivalve Scrobicularia plana (da Costa) settling in fine sand. Exposure to nominal TBT levels of 125 ng Sn l^-1 (actual range of 37 to 102 ng Sn l^-1, as analysed) resulted in substantial mortalities and negligible shell growth of individuals. More importantly, larval shell growth at the lowest concentration tested (i.e. nominal 50 ng Sn l^-1 or 14 to 32 ng Sn l^-1, as analysed) was significantly reduced (by a factor of 4) and grossly abnormal. In contrast, some control postlarvae had begun developing siphons as they attained a length of 600 to 700 m by the end of this test, thereby completing metamorphosis and becoming juveniles. The results are discussed in relation to the widespread disappearance of S. plana in North Atlantic European estuaries during the 1980s and the concurrent TBT pollution in their waters. It is inferred that TBT is likely to have contributed to the reported demise of clam populations by preventing the successful and timely progress of their pediveliger larvae.     

Preliminary study on the consumption of dissolved organic matter (exudates) of bacteria and phytoplankton by the marine bivalve Venus verrucosa

The utilisation of the exudates of bacteria (Lactobacillus sp.) and phytoplankton (Pavlova lutheri) by the clam Venus verrucosa Linné under identical experimental conditions was studied. 61.7% of the bacterial exudates were consumed but only 19.9% of the algal exudates. V. verrucosa would seem to assimilate bacterial products more easily than phytoplankton exudates. The study was performed in 1980.     

Chemoautotrophic symbionts and translocation of fixed carbon from bacteria to host tissues in the littoral bivalve Loripes lucinalis (Lucinidae)

Specimens of Loripes lucinalis (Lucinidae) living in reducing sediments were collected near a sewage outfall at low tide on the Moulin Blanc beach, Brest, France, from January to March 1987. Electron microscope studies revealed numerous Gram-negative-type bacteria in the gill cells. Ribulosebiphosphate carboxylase, a diagnostic enzyme of the Calvin-Benson cycle of CO₂-fixation was measured only in the gill extracts. Various tissues of L. lucinalis were examined for activity of APS reductase, (EC 1.8.99.2), ATP sulphurylase (EC 2.7.7.4) and rhodanese (EC 2.8.1.1), enzymes involved in sulphide oxidation. APS reductase was only found in symbiont-containing tissues, i.e., gills. These enzymatic studies characterise the symbionts as chemoautotrophic sulphide-oxidizing bacteria. Histoautoradiography demonstrated that part of the carbon dioxide fixed by symbiotic bacteria in the gills is translocated to symbiont-free tissues of the bivalve. The ultrastructure of the gill is detailed and a nomenclature based on established and new terminology is proposed to describe the various cellular types comprising the gill filament.     

Defensive secretions from the larvae ofApatania fimbriata (Pictet) (Trichoptera: Limnephilidae)

Summary When the larvae of the caddis flyApatania fimbriata (Pictet) are threatened, drops of fluid appear on their head capsules. The secretions are produced in a gland in the dorsal part of the prothorax. The neck region contains an eversible sac with numerous single setae, groups of 3 setae, or rows of setae on the surface. The secretion is released through two paris of orifices on the lateral sides of the sac. For the most part the secretion is composed of some 30 fatty acids, with the major components having 12–14 carbon atoms and up to 4 double bonds (approx. 1–2 µg secretion per specimen). Biotests with synthetic saturated acids (C₆–C₁₂) in a stream and in the laboratory demonstrated a paralysing effect on small invertebrate predators (Rhyacophila sp.,Plectrocnemia conspersa, Hydropsyche sp., larvae, all Trichoptera). In choice experiments,Rhyacophila sp. larvae preferred larvae ofAgapetus fuscipes andDrusus annulatus (Trichoptera) as food as compared withApatania fimbriata larvae. Larger predators, such asDinocras cephalotes (Insecta, Plecoptera) and the fishCottus gobio, did not discriminate betweenApatania fimbriata and other prey species. The use of fatty acids in defensive secretions is interpreted as an adaptation to the running water environment. They are effective repellents againstRhyacophila sp. larvae, the most important predator in the natural environment ofApatania larvae.     

Temperature tolerance of embryos and larvae of five bivalve species under simulated power plant entrainment conditions: a synthesis

Upwelling effects of subsurface water on phytoplankton growth were evaluated by 9 simulated culture experiments of coastal upwelling. Particular attention was paid to the effects of nutrient enrichment on the surface phytoplankton by the upwelling of nutrient rich subsurface water and of the exposure of the subsurface phytoplankters to surface radiation. The following are the results obtained: the lag period of phytoplankton growth was inversely related to water temperature; the maximum yield of phytoplankton was proportional to the amounts of available initial nutrients; the specific growth rates of phytoplankton were a function of both the initial nutrient concentrations and water temperature; and the maximum specific growth rate was simply proportional to water temperature. According to the relations found, a simple equation is presented for the estimation of phytoplankton growth in a given upwelling. Succession of species in the phytoplankton assemblage in upwelled water mass was also taken into consideration.     

Two new records of Planctosphaera larvae (Hemichordata: Planctosphaeroidea)

Two specimens of the rarely found Planctosphaera larva have been taken in the North Atlantic near Challenger Bank north of the Azores and in the North Equatorial Current, latitude 19°33 N, longitude 40°01 W.This research was supported by Grant 6027-X from the U.S. National Science Fonndation and is Contribution No. 2380 from the Woods Hole Oceanographic Institution.     

Association of filamentous epibacteria with Tubificoides benedii (Oligochaeta: Annelida)

A previously undescribed colonization of the marine oligochaete Tubificoides benedii (d'Udekum, 1855) from a sulfide-rich habitat by filamentous epibacteria was observed in 1984 and 1985. Electron microscopy showed that the bacteria were embedded in the cuticle of the posterior region of the worm. The association is apparently not pathogenic; possible forms of positive interaction are discussed.     

Size regulation in larvae of the crustacean Balanus eburneus (Cirripedia: Thoracica)

We tested the hypothesis that larval size in the acorn barnacle Balanus eburneus Gould (Cirripedia Thoracica) varies in relation to food availability. In March–November 1980, and March–July 1981, larvae were obtained from adult Balanus eburneus collected in the Newport River, North Carolina, USA. Carapace length and width of larvae reared at three different food concentrations were measured. Mean naupliar instar size was independent of food concentration. Mean size of the cypris instar increased with increasing food level. Greater cypris size could be attributed to increased food reserves in the preceding naupliar stage, and was coinciden with inmarked increase in metamorphic success. Variation in instar size remained constant or declined during naupliar development, but increased sharply at the molt to the cyprid. Naupliar size regulation involved: (1) conservation of a molt increment specific for each naupliar-naupliar molt, (2) an inverse relationship between premolt size and the molt increment during the first five naupliar instars, and (3) an increase in the precision of the molt increment at the molt to the sixth naupliar instar. Experimental evidence implies that size regulation in Balanus eburneus limits variation about a fixed final naupliar size (e.g. volume). Measurement of naupliar size, accumulated energy reserves, survival and development time, and cypris metamorphic success indicated that naupliar cuticular growth is the most conservative feature of larval development. The data suggest that maximum naupliar size is limited by escalating metabolic costs during development, while minimum naupliar size is limited by size-related feeding effectiveness.     

Eggs and early larvae of Thrissocles species (Engraulidae: Pisces)

Eggs of Thrissocles species are found in surface plankton in the Ernakulam Channel (Cochin Harbour)during February to May 1967. The eggs hatch within 24 h. Empty egg shells have characteristic apertures, through which the embryos have emerged; yolk is resorbed 36 h after hatching.Larvae (36 to 72 h groups) assemble at the lighted region of the aquaria during day-time and scatter to different levels at night. Larvae older than 72 h show no inclination to assemble as before. All larvae died between 96 and 110 h after hatching. Many batches of eggs were reared in the same medium, and all of them behaved as described. The results indicate that the right type of food was available in the aquaria for larvae up to a period of 72 h. The volume of water also appears to have a bearing upon the survival rate and longevity of the larvae since, in small aquaria, more larvae died at an early stage.     

Development and metamorphosis of the planktotrophic larvae of Rostanga pulchra (Mollusca: Nudibranchia)

Rostanga pulchra MacFarland, a small (1 to 2 cm) dorid nudibranch, lays an average of 7000 eggs in the laboratory during a period of 30 days in the summer. The veligers hatch 15 to 16 days after oviposition and it takes another 35 to 40 days to become competent for metamorphosis at a temperature of 10° to 15°C. Larval cultures were maintained initially at a concentration of 500 veligers per 100 ml of filtered sea water (antibiotics added). During the planktotrophic phase of development, the veliger grows from 150 to 300 m in shell length. Although the veligers are generalists in their food preference, the best result (faster growth) was achieved by feeding them with a combination of Monochrysis lutheri and Isochrysis galbana. The concentration of food cells was kept at 10⁴ cells per ml of culture media and was supplied every 2 to 3 days. A veliger which is competent to metamorphose is identifiable morphologically by its propodium, eyespots, rhinophores, and spiculated dorsal papillae. The entire metamorphic process lasts 24 h when a suitable substrate such as the food sponge Ophlitaspongia pennata is provided. The competent veliger is able to delay metamorphosis for at least 3 weeks. Juveniles were kept in the laboratory for 70 days and, during this period, grew to a length of 4.5 mm.