d-Alanine metabolism in the lucinid clam Lucinoma aequizonata

The chemoautotrophic symbiont-bearing clam Lucinoma aequizonata contains very high levels of free d-alanine in all tissues. The possible sources for this amino acid and its involvement in the clams' metabolism were investigated. Very low levels of d-alanine (generally below 1 mol·l^-1) were measured in the sediment porewaters from the habitat of the clams. Experiments with ¹⁴C-labeled tracers demonstrate an active metabolism of d-alanine in the clams rather than a role as inert waste product. d-alanine is metabolized at about 0.12 mol·g fw^-1·h^-1. Label from aspartate, but not glucose and CO₂, is incorporated into d-alanine. Incubation with labeled d-alanine did not result in formation of radioactive l-alanine. Tests for alanine racemase (EC 5.1.1.1) and d-amino acid oxidase (EC 1.4.3.3.) did not show activity in either gill, i.e. symbiont and host, or foot tissue. d-Alanine amino transferase (EC 2.6.1.b.) was demonstrated in gill and foot tissues. Two sources for d-alanine are proposed: a degradation of cell walls of symbiotic bacteria and production by the host using a d-specific alanine transaminase.Abbreviations aa amino acid(s) - fw fresh weight - HPLC high-performance liquid chromatography - MBH methyl benzethonium hydroxyde - NAC N-acetyl-l-cysteine - OPA ortho-phthaldialdehyde - TCA tricarbonic acid     

A GIANT LUCINID BIVALVE FROM THE EOCENE OF JAMAICA – SYSTEMATICS, LIFE HABITS AND CHEMOSYMBIOSIS (MOLLUSCA: BIVALVIA: LUCINIDAE)

Abstract:  The giant bivalve Lucina megameris Dall, 1901 , from the late Eocene White Limestone Group of Jamaica and by far the largest known species of the family Lucinidae, is placed in a new genus Superlucina . Apart from its large size, with a shell height exceeding 310 mm, it is distinguished from other genera, such as Pseudomiltha and Eomiltha by external shell characters and the extremely long and narrow, anterior adductor muscle scar. Features preserved on internal moulds suggest that, in common with living Lucinidae, S. megameris was chemosymbiotic with sulphide – oxidizing bacteria housed in the gills. Palaeoenvironmental evidence suggests a habitat in oligotrophic, shallow waters, probably in seagrass beds, with an associated molluscan fauna including large cardiids that may have been photosymbiotic. Superlucina is considerably larger than any living lucinid that range in size from 3 to 150 mm with most encompassed within 5–30 mm. From the Jurassic onwards, a few other large lucinids are known from cold seep sites, with several other records from possible shallow water seagrass beds.     

Universal primers and PCR of gut contents to study marine invertebrate diets

Determining the diets of marine invertebrates by gut content analysis is problematic. Many consumed organisms become unrecognizable once partly digested, while those with hard remains (e.g. diatom skeletons) may bias the analysis. Here, we adapt DNA-based methods similar to those used for microbial diversity surveys as a novel approach to study the diets of macrophagous (the deep-sea amphipods Scopelocheirus schellenbergi and Eurythenes gryllus) and microphagous (the bivalve Lucinoma aequizonata) feeders in the deep sea. Polymerase chain reaction (PCR) in conjunction with 'universal' primers amplified portions of the mitochondrial cytochrome c oxidase I (COI) gene for animals ingested by S. schellenbergi and E. gryllus and the 18S rRNA gene for lesser eukaryotes ingested by L. aequizonata. Amplified sequences were combined with sequences from GenBank to construct phylogenetic trees of ingested organisms. Our analyses indicate that S. schellenbergi, E. gryllus and L. aequizonata diets are considerably more diverse than previously thought, casting new light on the foraging strategies of these species. Finally, we discuss the strengths and weaknesses of this technique and its potential applicability to diet analyses of other invertebrates.     

Report of a cohesive gelatinous egg mass produced by a tropical marine bivalve

Abstract. Gelatinous egg masses are common in a number of animal phyla. However, they are virtually unknown in marine bivalves, with structures that could be thought of as gelatinous egg masses being reported for only five species. We describe the gelatinous egg mass and intracapsular development in the tropical lucinid Phacoides pectinatus. The embryos developed within individual capsules embedded in a large flimsy, spherical mass. Swimming veligers hatch at 198 μm shell length. They did not feed, settled within several days of hatching, and metamorphosis was completed within 2 weeks of hatching. Gelatinous egg masses might be detected in members of more lucinid species if studies of development included field or in vivo observations of reproduction in addition to producing embryos by stripping the gonads.     

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

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

Chemosymbiotic bivalves and stable carbon isotopes indicate hydrocarbon seepage at four unusual Cenozoic fossil localities

Four unusual Cenozoic fossil localities are identified here as ancient hydrocarbon seep sites using palaeontological, petrological, and stable carbon isotope data. Late Eocene carbonate-cemented sandstone bodies in the Wagonwheel Mountains in California were previously suspected to represent ancient hydrocarbon seep sites, but the relatively high δ¹³C_carbonate values (−12.2 to −5.0‰) suggest oil rather than methane seepage. The Oligocene fauna of the Elmira asphalt mine, Cuba, was previously interpreted as a mix of freshwater and marine taxa, but all species are here identified as belonging to marine groups, including the bivalve families Lucinidae and Vesicomyidae, whose extant members live largely in symbiosis with chemoautotrophic endosymbionts. A carbonate concretion from this site showed δ¹³C_carbonate values as low as −32.2‰, which most likely indicates methane seepage. A previously unpublished Oligocene fossil locality in Atlantico, northern Colombia, is dominated by large solemyid, mytilid, lucinid, and vesicomyid bivalves, which most likely lived with chemotrophic endosymbionts. Seepage of biogenic methane without a significant contribution of thermogenic methane is indicated by δ¹³C_carbonate values as low as −51.3‰. We confirm that the Pleurophopsis -dominated sites of the Heath Shale and Lomitos Chert in northern Peru are ancient seep sites, although the previous identification of Pleurophopsis peruviana as a vesicomyid is doubtful.     

Embryonic development and endosymbiont transmission mode in the symbiotic clam Lucinoma aequizonata (Bivalvia: Lucinidae)

Summary

Lucinoma aequizonata is a large lucinid clam which lives in reducing mud around 500 m deep. Adults harbor intracellular chemoautotrophic sulfur-oxidizing bacteria in specialized gill cells called bacteriocytes. The embryonic and early larval development of L. aequizonata is described by using light and scanning electron microscopy. Gametes were obtained by injection of 0.2ml of 4 mM serotonin solution in seawater into the posterior adductor muscle. The oocytes, 200 μm in diameter, are surrounded by a glycoprotein capsule which gives to the egg a total diameter of 500μm. The development which occurs at 10°C is slow. The first polar body is detected 2.5 h after contact between sperm and oocytes (T_o+2.5 h), and the first cleavage begins 10 h later (T_o+12.5 h). The following successive cleavages produce a nonciliated morula, then a ciliated gastrula which begins to rotate within the egg-capsule at T_o+4.5 days. At this stage, the first shell pellicle appears on the dorsal side of the embryo. At T₀+8 days, the trochophore larvae develop discrete ciliary bands which constitute the prototroch. Typical straight-hinge veligers, D-shaped larvae, hatch from the egg-capsule 12 days after fertilization. The newly hatched larvae are 240 μm in length and 200μm in height, and the straight hinge 150μm long. To elucidate the symbiont transmission mode, two symbiont-specific primers were designed and used in amplifications by PCR. This primer set was unsuccessful in amplifying symbiont DNA targets from mature gonads, spawned oocytes, eggs, and veligers whereas successful amplifications were obtained from symbiont-containing gill tissues. These data rule out the vertical transmission mode and strongly suggest that the symbionts are environmentally transmitted to the new host generation in L. aequizonata as for all tropical lucinids examined to date.