The reproductive cycle in a typical estuarine mollusc Nausitora hedleyi Schepman based on a study of the Gonad Index

1. Previous work on the methods employed for the determination of the breeding season of shipworms is briefly reviewed.
2. The method adopted for studying the reproductive cycle by using the “gonad index” is described.
3. The reproductive cycle of Nausitora hedleyi is described in detail based on a study of the gonad index of different sexes collected at monthly intervals from the estuarine environment of Cochin harbour.
4. The fact that breeding is restricted as marked by seasonal activity is shown from the size and activity of the gonad during the different months of the year.
5. The environment, and the hydrographic conditions prevailing in the habitat of N. hedleyi in the Cochin harbour are described.

     

Bacterial symbiont transmission in the wood-boring shipworm Bankia setacea (Bivalvia: Teredinidae)

The Teredinidae (shipworms) are a morphologically diverse group of marine wood-boring bivalves that are responsible each year for millions of dollars of damage to wooden structures in estuarine and marine habitats worldwide. They exist in a symbiosis with cellulolytic nitrogen-fixing bacteria that provide the host with the necessary enzymes for survival on a diet of wood cellulose. These symbiotic bacteria reside in distinct structures lining the interlamellar junctions of the gill. This study investigated the mode by which these nutritionally essential bacterial symbionts are acquired in the teredinid Bankia setacea. Through 16S ribosomal DNA (rDNA) sequencing, the symbiont residing within the B. setacea gill was phylogenetically characterized and shown to be distinct from previously described shipworm symbionts. In situ hybridization using symbiont-specific 16S rRNA-directed probes bound to bacterial ribosome targets located within the host gill coincident with the known location of the gill symbionts. These specific probes were then used as primers in a PCR-based assay which consistently detected bacterial rDNA in host gill (symbiont containing), gonad tissue, and recently spawned eggs, demonstrating the presence of symbiont cells in host ovary and offspring. These results suggest that B. setacea ensures successful inoculation of offspring through a vertical mode of symbiont transmission and thereby enables a broad distribution of larval settlement.     

Occurrence and settlement of the common shipworm Teredo navalis (Bivalvia: Teredinidae) in Bremerhaven harbours, northern Germany

The shipworm Teredo navalis L. is a xylophagous bivalve mollusc (Bivalvia: Teredinidae) with a long record of being very destructive to wooden ships and harbour buildings. It has been reported from numerous sites at the coasts of both the North and Baltic Seas since the eighteenth century. Here, we document for the first time the occurrence of live adult T. navalis in the harbours of Bremerhaven (Weser estuary, northern Germany). From August to December 1998, various wooden structures (fir floating fenders and pier posts, oak piles) from seven stations in different docks of two harbours (überseehafen, Fischereihafen) were investigated for the presence and density of live specimens and burrows of T. navalis. The settlement of larval shipworms was studied by exposing experimental fir panels 0.06 m² in size at 20 stations at water depths between 1 and 2 m for periods of 4 months between July and November. In addition, hydrographic profiles (0–8 m water depth) were obtained at 17 stations in five docks once every month from August to December. Live adult shipworms were found in both fir floating fenders and oak piles at four stations. The largest specimen found was 250 mm long. Shipworm burrows were detected at five stations in almost every wooden structure investigated but their abundances differed significantly: Maximum values were >10,000 m^–2 in fir floating fenders, 4,600 m^–2 in oak piles and 200 m^–2 in fir pier posts. Actual shipworm infestation was detected at three of 16 stations in the exposed fir panels (1–3 burrow holes per panel). Water temperatures and salinities varied considerably during the 4-month investigation period. Temperatures decreased from 19.9°C in August to 0.7°C in December. Salinities ranged from 17.6 in August to 1.1 in November, but only at two lock stations during November and December did value drop below 5, which is regarded as the lethal limit for the larvae of this euryhaline teredinid species. We conclude that T. navalis encounters favourable conditions for growth and reproduction in the harbours of Bremerhaven, at least during summer and autumn, and is a common element of the harbour ecosystem. Therefore, a persistent infestation of all wooden structures after a relatively short period of time seems to be highly probable. Electronic Publication     

Location of glycogen in spermatids and spermatozoa of the shipworm,Bankia australis (Teredinidae,Bivalvia, Mollusca)

The periodic acid-thiocarbohydrazide-silver proteinate technique and alpha-amylase digestion were used to locate glycogen in the spermatids and sperms of the bivalve Bankia australis. Glycogen was found in the middle piece and around the acrosome in spermatozoa, apparently randomly scattered throughout the cytoplasm of young spermatids, and in a cytoplasmic bead in old spermatids.     

The broadcast spawning Caribbean shipworm,Teredothyra dominicensis (Bivalvia,Teredinidae), has invaded and become established in the eastern Mediterranean Sea

Teredinids, commonly referred to as shipworms, are wood-boring bivalves estimated to cause over one billion dollars’ worth of damage to submerged wooden structures per annum. This paper reports the detection and establishment of the Caribbean shipworm Teredothyra dominicensis (Bivalvia, Teredinidae) in the eastern Mediterranean Sea. Identification was confirmed using an integrative taxonomical approach combining morphology, morphometry and molecular markers (COI-5P and 18S), thus improving both the taxonomic resolution and tractability of this invasive species. Sequence comparisons between indigenous Caribbean and Mediterranean specimens were at least 99 % identical. Wooden panels placed at the site of discovery were infested exclusively by T. dominicensis with specimens of varying size and age, indicating multiple settlement events and the presence of breeding populations in the region. Anatomical and behavioural observations confirm the species as a broadcast spawner with larvae undergoing planktotrophic development, thus distribution range is potentially extensive. Of the possible introduction vectors, transport via ballast water is proposed as the most likely. The establishment of breeding populations of a tropical teredinid in the Mediterranean is of considerable concern as tropical species are particularly destructive and degrade wood more rapidly than the species currently found in the region. This threat is likely to increase in severity due to global warming, as increases in temperature and salinity may lead to an increase in the distribution range, development rate and boring activity of teredinids.     

Bacterial Symbiont Transmission in the Wood-Boring Shipworm Bankia setacea (Bivalvia: Teredinidae)

The Teredinidae (shipworms) are a morphologically diverse group of marine wood-boring bivalves that are responsible each year for millions of dollars of damage to wooden structures in estuarine and marine habitats worldwide. They exist in a symbiosis with cellulolytic nitrogen-fixing bacteria that provide the host with the necessary enzymes for survival on a diet of wood cellulose. These symbiotic bacteria reside in distinct structures lining the interlamellar junctions of the gill. This study investigated the mode by which these nutritionally essential bacterial symbionts are acquired in the teredinid Bankia setacea. Through 16S ribosomal DNA (rDNA) sequencing, the symbiont residing within the B. setacea gill was phylogenetically characterized and shown to be distinct from previously described shipworm symbionts. In situ hybridization using symbiont-specific 16S rRNA-directed probes bound to bacterial ribosome targets located within the host gill coincident with the known location of the gill symbionts. These specific probes were then used as primers in a PCR-based assay which consistently detected bacterial rDNA in host gill (symbiont containing), gonad tissue, and recently spawned eggs, demonstrating the presence of symbiont cells in host ovary and offspring. These results suggest that B. setacea ensures successful inoculation of offspring through a vertical mode of symbiont transmission and thereby enables a broad distribution of larval settlement.     

The fine structure of the oocyte of Bankia australis (Teredinidae,Bivalvia) before and after fertilization

The fine structure of the oocyte of Bankia australis is compared with that of other bivalve oocytes. It was observed that following fertilization, the microvilli changed their spatial organisation and behaviour towards sperm, the cortical granules disappeared in regions of high concentrations of supernumerary sperm, and the mitochondria apparently stated to divide.     

Distribution, settlement, and growth of first-year individuals of the shipworm Teredo navalis L. (Bivalvia: Teredinidae) in the Port of Rotterdam area, the Netherlands

During the period 2004-2008 the distribution, settlement, and growth of first-year shipworms (Teredo navalis L., 1758) was studied by exposing fir and oak panels in the Port of Rotterdam area, which is situated in the Rhine-Meuse estuary in the Netherlands and covers the complete salinity gradient. Shipworms were found yearly in the western large polyhaline harbours. On only a few occasions were they were found in harbours that showed large seasonal and daily fluctuations in salinity. In 2006 the shipworm was found in fir panels 20 km upstream from the polyhaline harbours, demonstrating their ability to travel with the tidal currents over considerable distances and to settle once the abiotic conditions become favourable. Although the water temperatures allowed them to breed from April until November, infestations were not found before September, and from the size of the animals in the panels it was concluded that in the Port of Rotterdam area they spawned from August until the end of November. The settlement height was negatively correlated with the distance of the panels to the sea floor. In the first season after settlement they showed a substantial growth rate of 0.18 cm day⁻¹. The longest shipworm found measured 36.8 cm after 4-5 months of growth after settlement. Infestations and growth were lower in oak than in fir wood. In 2006 the maximum consumption of wood by individuals settled in the same year in panels at the bottom accounted for 12.4%. Shell size and body length of the animal after the first season of growth showed a significant positive logarithmic relation. In both 2006 and 2007 a similar relation between the average boring tube diameter and the length of the animals was found. Lower river discharges leading to salinisation of the eastern part of the Port of Rotterdam area create conditions favourable for the shipworm, with serious consequences for the piles upon which the quays are built.     

The influence of salinity in the domoic acid effect on estuarine phytoplankton communities

Toxic species of the diatom genus Pseudo-nitzschia, observed worldwide from coastal waters to the open ocean, produce the neurotoxin domoic acid (DA). DA is an important environmental and economic hazard due to shellfish contamination with subsequent effects on higher trophic levels. Previous research has demonstrated that, among other environmental factors, salinity influences the abundance and toxicity of Pseudo-nitzschia. In this study, the environmental factors driving the growth of Pseudo-nitzschia and the production of dissolved DA (dDA) in North Inlet estuary were examined. The effect of salinity on the growth inhibition of phytoplankton induced by the initial presence as well as by an addition of dDA was also assessed. Initially, the diatom abundance was negatively correlated with the abundance of Pseudo-nitzschia and with the concentration of dDA. With the addition of a concentrated solution of dDA, the percent inhibition of cryptophytes and diatoms was significantly correlated with salinity and suggested a higher sensitivity to dDA at extreme salinities. These results emphasize the importance of salinity in assessing the properties of DA and potentially of other phycotoxins on phytoplankton.     

Microbial Distribution and Abundance in the Digestive System of Five Shipworm Species (Bivalvia: Teredinidae)

Marine bivalves of the family Teredinidae (shipworms) are voracious consumers of wood in marine environments. In several shipworm species, dense communities of intracellular bacterial endosymbionts have been observed within specialized cells (bacteriocytes) of the gills (ctenidia). These bacteria are proposed to contribute to digestion of wood by the host. While the microbes of shipworm gills have been studied extensively in several species, the abundance and distribution of microbes in the digestive system have not been adequately addressed. Here we use Fluorescence In-Situ Hybridization (FISH) and laser scanning confocal microscopy with 16S rRNA directed oligonucleotide probes targeting all domains, domains Bacteria and Archaea, and other taxonomic groups to examine the digestive microbiota of 17 specimens from 5 shipworm species (Bankia setacea, Lyrodus pedicellatus, Lyrodus massa, Lyrodus sp. and Teredo aff. triangularis). These data reveal that the caecum, a large sac-like appendage of the stomach that typically contains large quantities of wood particles and is considered the primary site of wood digestion, harbors only very sparse microbial populations. However, a significant number of bacterial cells were observed in fecal pellets within the intestines. These results suggest that due to low abundance, bacteria in the caecum may contribute little to lignocellulose degradation. In contrast, the comparatively high population density of bacteria in the intestine suggests a possible role for intestinal bacteria in the degradation of lignocellulose.     

Mytilus galloprovincialis (Mollusca: Bivalvia) in a warm-temperate South African estuarine embayment

The distribution, population structure, condition and gonad indices, and the attachment strengths of the invasive Mediterranean mussel Mytilus galloprovincialis are described from the Knysna Estuary, South Africa. Mussels were found in the rocky intertidal at the mouth of the estuary and on all man-made hard substrata as far as 12?km from the mouth in 2013. However, mussel beds were found only in the lower reach of the estuary, where mussel densities were as high as 120 per 0.01?m². When compared to mussels from the embayment regions of the estuary, those at the estuary mouth, where wave action was high, were generally smaller, had significantly lower condition and gonad indices, and greater attachment strength. This suggests that mussels within the sheltered regions of the estuary invest more energy in growth and reproduction, and that this is one reason for their success as an invasive species in the Knysna estuarine embayment.     

Physiological and biochemical energetics of larvae of Teredo navalis L. and Bankia gouldi (Bartsch) (Bivalvia : Teredinidae)

The biochemical composition of larvae of Teredo navalis L. and Bankia gouldi (Bartsch) (Bivalvia: Teredinidae) was examined throughout larval development at 23°C and 30–32%. salinity in the presence of the phytoplankton food Isochrysis aff. galbana (clone T-ISO), during a delay of metamorphosis in the presence of food but absence of a wood substratum and during periods of enforced starvation. Newly released Teredo navalis larvae had a mean length (L) and height (H) of 89.3 and 76.1 μm respectively. The first appearance of pediveliger larvae at 212.1 μm L and 230.0 μm H occurred 27 days after release. Larval dry weight increased from 0.29 μg to 1.96 μg during this period. Newly formed straight hinge larvae of Bankiagouldi had dimensions of 62.8 μm L and 49.8 μm H. Metamorphically competent B. gouldi larvae had dimensions of 230.0 μm L and 282.9 μm H and were first observed 20 days after fertilization. Larval dry weight increased from 0.06 μg to 2.20 μg during this period. During enforced delay of metamorphosis the ash-free dry weight of Teredo navalis larvae decreased whereas the ash free dry weight of Bankia gouldi larvae increased. During the early period of shelled larval development both species showed similar decreases in lipid, protein and carbohydrate levels (μg·mg dry weight^?1); however, this was reflected in a decrease in biochemical content (μg·larva^?1) only in Teredo navalis. During enforced starvation the major proportion of both the weight and caloric losses were due to protein. Lipid also contributes significantly to these losses whereas the contribution of carbohydrate was small. Larval oxygen consumption rates were determined directly by manometry and indirectly by estimates of decrease in caloric content during periods of enforced starvation. Direct and indirect determinations for T. navalis are described by the relationships R = 1.16 W^1.05 and R = 0.98 W^1.24 respectively where R is the respiration rate in nl O₂ · larva^?1 · h^?1 and W is dry weight inclusive of shell in μg. Direct and indirect determinations for Bankia gouldi are described by the relationships R = 1.37 W^1.25 and R = 1.81 W^1.25 respectively. When data for both assay procedures are combined for each species the relationships R = 1.10 W^1.07 and R = 1.44 W^1.18 are obtained for Teredonavalis and Bankia gouldi respectively.     

Coexistence of multiple proteobacterial endosymbionts in the gills of the wood-boring Bivalve Lyrodus pedicellatus (Bivalvia: Teredinidae)

Wood-boring bivalves of the family Teredinidae (commonly called shipworms) are known to harbor dense populations of gram-negative bacteria within specialized cells (bacteriocytes) in their gills. These symbionts are thought to provide enzymes, e.g., cellulase and dinitrogenase, which assist the host in utilizing wood as a primary food source. A cellulolytic, dinitrogen-fixing bacterium, Teredinibacter turnerae, has been isolated from the gill tissues of numerous teredinid bivalves and has been proposed to constitute the sole or predominant symbiont of this bivalve family. Here we demonstrate that one teredinid species, Lyrodus pedicellatus, contains at least four distinct bacterial 16S rRNA types within its gill bacteriocytes, one of which is identical to that of T. turnerae. Phylogenetic analyses indicate that the three newly detected ribotypes are derived from gamma proteobacteria that are related to but distinct (>6.5% sequence divergence) from T. turnerae. In situ hybridizations with 16S rRNA-directed probes demonstrated that the pattern of occurrence of symbiont ribotypes within bacteriocytes was predictable and specific, with some bacteriocytes containing two symbiont ribotypes. However, only two of the six possible pairwise combinations of the four ribotypes were observed to cooccur within the same host cells. The results presented here are consistent with the existence of a complex multiple symbiosis in this shipworm species.     

The growth rates of the estuarine mollusc Musculista arcuatula Yamamoto and Habe (Bivalvia: Mytilidae)

The pattern of growth of the total population of Musculista arcuatula over a period of thirteen months was built up plotting length-percentage frequency histograms each month. From the shifting of the modes it was noticed that the increase in size from November to December was 2 mm, the growth during December to January 3 mm. In February the growth was nil. In March the size increasing by 6 mm and in April by 1 mm, when mortality of the largest size groups started to get completed in May. In June a conspicuous mode appeared at 9 mm, grew 1 mm in July, 2 mm in August and 3 mm in September. Growth remained stationary in October and November. The rate of increase in length was found to follow exponential law. The growth rate of this bivalve goes on increasing at a steady pace, the rate being 1.086 mm per month. The mean size of the females was found to be significantly larger than that of the males. The proportion of the males in the size group 3 mm to 10 mm was found to be significantly larger than in the entire population. But in the larger size groups there was no difference in the number of males and females and the proportion was almost 1:1. Throughout the year only one age class was recognizable in the natural population. There appears to be an excessive post reproductive mortality of adults rendering the population during most of the year of a single age class recruited through the various months of spawning. The maximum longevity was found to be at the most 18 months.     

Extensive variation in intracellular symbiont community composition among members of a single population of the wood-boring bivalve Lyrodus pedicellatus (Bivalvia: Teredinidae)

Shipworms (wood-boring bivalves of the family Teredinidae) harbor in their gills intracellular bacterial symbionts thought to produce enzymes that enable the host to consume cellulose as its primary carbon source. Recently, it was demonstrated that multiple genetically distinct symbiont populations coexist within one shipworm species, Lyrodus pedicellatus. Here we explore the extent to which symbiont communities vary among individuals of this species by quantitatively examining the diversity, abundance, and pattern of occurrence of symbiont ribotypes (unique 16S rRNA sequence types) among specimens drawn from a single laboratory-reared population. A total of 18 ribotypes were identified in two clone libraries generated from gill tissue of (i) a single specimen and (ii) four pooled specimens. Phylogenetic analysis assigned all of the ribotypes to a unique clade within the gamma subgroup of proteobacteria which contained at least five well-supported internal clades (phylotypes). By competitive quantitative PCR and constant denaturant capillary electrophoresis, we estimated the number and abundance of symbiont phylotypes in gill samples of 13 individual shipworm specimens. Phylotype composition varied greatly; however, in all specimens the numerically dominant symbiont belonged to one of two nearly mutually exclusive phylotypes, each of which was detected with similar frequencies among specimens. A third phylotype, containing the culturable symbiont Teredinibacter turnerae, was identified in nearly all specimens, and two additional phylotypes were observed more sporadically. Such extensive variation in ribotype and phylotype composition among host specimens adds to a growing body of evidence that microbial endosymbiont populations may be both complex and dynamic and suggests that such genetic variation should be evaluated with regard to physiological and ecological differentiation.     

Extensive Variation in Intracellular Symbiont Community Composition among Members of a Single Population of the Wood-Boring Bivalve Lyrodus pedicellatus (Bivalvia: Teredinidae)

Shipworms (wood-boring bivalves of the family Teredinidae) harbor in their gills intracellular bacterial symbionts thought to produce enzymes that enable the host to consume cellulose as its primary carbon source. Recently, it was demonstrated that multiple genetically distinct symbiont populations coexist within one shipworm species, Lyrodus pedicellatus. Here we explore the extent to which symbiont communities vary among individuals of this species by quantitatively examining the diversity, abundance, and pattern of occurrence of symbiont ribotypes (unique 16S rRNA sequence types) among specimens drawn from a single laboratory-reared population. A total of 18 ribotypes were identified in two clone libraries generated from gill tissue of (i) a single specimen and (ii) four pooled specimens. Phylogenetic analysis assigned all of the ribotypes to a unique clade within the γ subgroup of proteobacteria which contained at least five well-supported internal clades (phylotypes). By competitive quantitative PCR and constant denaturant capillary electrophoresis, we estimated the number and abundance of symbiont phylotypes in gill samples of 13 individual shipworm specimens. Phylotype composition varied greatly; however, in all specimens the numerically dominant symbiont belonged to one of two nearly mutually exclusive phylotypes, each of which was detected with similar frequencies among specimens. A third phylotype, containing the culturable symbiont Teredinibacter turnerae, was identified in nearly all specimens, and two additional phylotypes were observed more sporadically. Such extensive variation in ribotype and phylotype composition among host specimens adds to a growing body of evidence that microbial endosymbiont populations may be both complex and dynamic and suggests that such genetic variation should be evaluated with regard to physiological and ecological differentiation.     

Assessment of the influence of confounding factors (weight,salinity) on the response of biomarkers in the estuarine polychaete Nereis diversicolor

The influences of salinity and body size on biochemical (activities of glutathione-S-transferase, lactate dehydrogenase (LDH), acetylcholinesterase and digestive enzymes amylase and CMCase), physiological (feeding and egestion rates, energy reserves) and behavioural (burrowing speed) biomarkers were examined in the infaunal polychaete Nereis diversicolor. Only a few biomarkers were affected, including increased egestion rate and activities of CMCase and LDH at higher salinity, and higher egestion rate in larger worms. These findings reinforce the status of N. diversicolor as a robust sentinel species for estuaries which are environments that are particularly productive but also particularly at risk.     

Molluscan assemblage from a tropical intertidal estuarine sand-mud flat, Gulf of Nicoya, Pacific, Costa Rica (1984-1987)

The availability of data sets covering more than a year is scarce for tropical environments. Advances in hardware and software speed-up the re-analysis of old data sets and facilitates the identification of hidden data patterns. From February 1984 to April 1987 (49 sampling dates), core samples (17.7cm2, 15cm deep) were collected at low tide at a sand-mud flat in the mid upper Gulf of Nicoya estuary, Pacific, Costa Rica. Predator exclusion experiments (cages 0.5 x 0.5 x 0.2m, galvanized wire, mesh size 5mm), were conducted at the site in 1985 (dry and rainy seasons sets). Samples were preserved with 5% buffered formalin in sea water stained with Rose Bengal, and washed after 24 hours on a 500 micron mesh sieve. The 1 120 cores yielded a total of 112 morphological species of which the mollusks were represented by 23 species, and included the bivalves Tellina rubescens, Tagelus bourgeoisae, Dosinia dunkeri and Leukoma asperrima, and the gastropods, Natica unifasciata, Nassarius luteostomus, Costoanachis rugosa and Turbonilla sp. The 23 species are indicative of a relatively rich sedimentary molluscan fauna. T. bourgeoisae had a seasonal oscillation, with higher abundances during the rainy seasons. T. rubescens was not seasonal, but presented an oscillation with peaks at about 1.5 year intervals. Many empty shells of Cosmioconcha modesta, lower number of N. luteostomus and a few of T. rubescens were found with boreholes by the predatory snail N. unifasciata. T. rubescens was not significantly more abundant inside or outside cages. T. bourgeoisae showed a significant increase within the caged areas. The numerical fluctuations of the mollusks became more important during the rainy season of 1985. Red tide outbreaks in the Gulf of Nicoya in 1985 may have had an impact on the molluscan populations.