Environmental contaminants influencing immunefunction in marine bivalve molluscs

The increased observation of pollution induced disease conditions in marine organisms has led to a growing interest on the effects of environmental contaminants on the immune system. Most studies on modulation of the immune system in bivalves by pollutants have concentrated on the effects of heavy metals and polycyclic aromatic hydrocarbons (PAHs). The current literature on contaminant effects on specific components of the bivalve immune system is reviewed together with the effects on susceptibility to infection. Data are presented showing the effects on immune parameters of exposure to Vibrio tubiashi following pre-exposure to copper or cadmium. Mussels exposed to cadmium for 7 days followed by 7 days exposure to V. tubiashi demonstrated significantly higher numbers of circulating haemocytes compared with non-Vibrio-exposed groups. Similar experiments conducted with copper exposure for both 7 days and 7 weeks followed by V. tubiashi for 7 days demonstrated a significant decrease in the percentage of circulating eosinophils compared with basophilic cells for both short and long term exposures. The intracellular release of superoxide (NBT reduction) by haemocytes was stimulated in Vibrio-challenged mussels with no copper pre-exposure but was significantly reduced in mussels pre-exposed to 0·2 ppm of copper for 7 weeks. The mortalities for the copper experiments showed increased levels with increasing copper concentration and were consistently higher in the V. tubiashi challenged mussels which had also been exposed to copper.     

Protein gradients in byssal threads of some marine bivalve molluscs

Many marine bivalve molluscs produce byssal threads for attachment to solid substrata. Small (less than 10 mm) consecutive sections of the byssal threads of Mytilus edulis, M. californianus, Geukensia demissa, Atrina vexillum, and A. rigida were analyzed by amino acid analysis to determine if chemical composition remains constant as a function of location in thread segments. Nonlinear longitudinal protein gradients, probably involving collagen and an elastic protein, were found in the Mytilus species. In these, collagen peaks in the distal third of the thread. In Geukensia and the Atrina species, although the two differed greatly in composition, there is a clear nonvariability in composition of the thread within each species as a function of location in the thread. The adhesive plaque at the tip of the thread of all species examined differs substantially in composition from the remainder of the thread. Protein gradients in the threads of some bivalves may reflect specific adaptations evolved to respond to exposed habitats in high-energy environments.     

The marine bivalve molluscs pathogen Vibrio neptunius produces the siderophore amphibactin,which is widespread in molluscs microbiota

Amphiphilic siderophores, including amphibactins, are the most abundant siderophores in oceans. Genes putatively encoding the amphibactin system were proposed in some bacteria and homologues of these genes are particularly abundant in multiple bacterial lineages inhabitant of low-iron seawater. However, since no defective mutant strains in any of these genes were studied to date, their role in amphibactin synthesis or uptake was not demonstrated. In this work, an in silico analysis of the genome of the mollusc pathogen Vibrio neptunius leads us to identify a gene cluster (denoted absABDEF) that is predicted to encode an amphibactin-like siderophore and several mutant strains unable to synthesize or use siderophores were constructed. The results showed that genes absABDEF are required for amphibactin synthesis. A comparative chemical analysis of V. neptunius wild type and biosynthesis mutants allowed us to identify a mixture of nine amphibactin forms produced by this bacterium. In addition, the gene abtA is predicted to encode the ferri-amphibactin outer membrane transporter. The prevalence of the amphibactin system in bivalve hemolymph microbiota was also studied. We found that the amphibactin system is widespread in hemolymph microbiota including both commensal and pathogenic bacterial species. Thus, its contribution to bacterial fitness must be more related to environmental persistence than to pathogenicity.     

Degradation of bacterial lipopolysaccharides by digestive-gland extracts of marine bivalve molluscs

• 1.1. Crude digestive gland extract (DGE) of 5 species of marine bivalve mollusc had glycosidase activity against lipopolysaccharides (LPS) of gram-negative bacteria.
• 2.2. Most of these extracts, after ammonium sulphate precipitation, had higher glycosidase activity than commercial Helix pomatia gut juice.
• 3.3. Inorganic phosphate was also released from LPS by the various DGE but the lipid moiety of LPS appeared to be resistant to attack except by a DGE from Cerastoderma edule, which released small quantities of only non-hydroxylated fatty acids.

     

Phenotypic flexibility and physiological tradeoffs in the feeding and growth of marine bivalve molluscs

Bivalve molluscs have a highly plastic feeding and growth physiology.The increasing availability of families artificially selectedfor faster growth has enabled physiological experiments to investigatethe genetic basis for variable rates of growth. Fast growthis achieved by a combination of increased rates of feeding,reduced metabolic rates and lower metabolic costs of growth.In at least one species there is a trade-off between growthin protein and the storage of lipids that are utilized in gametogenesis.Energy requirements for maintenance are also higher in slow-growingindividuals. Reduced costs of growth are due in part to increasedefficiencies of protein turnover. Nevertheless, high proteinturnover (and therefore high metabolic cost) may benefit fitnessin the later stages of gametogenesis. Faster feeding rates donot impair flexibility in feeding behavior which compensatesfor changes in the food environment. Both inter- and intra-speciesdifferences in feeding behavior are evident and suggest possibleconstraints imposed by faster feeding on the efficiency of selectionbetween food particles of different nutritional value.     

Hox and paraHox genes in bivalve molluscs

In this study, we sought the presence and analysed the sequences of the Hox and ParaHox genes in bivalve molluscs. The clustered Hox genes play a central role in anterior-posterior axial patterning in bilaterian metazoa, whereas the ParaHox gene cluster is a paralogue (evolutionary sister) of the Hox cluster.Using polymerase chain reaction (PCR)-based approaches, we isolated nine different sequences in five species belonging to three of the main bivalve subclasses: Ensis ensis and Tapes philippinarum (Heterodonta), Pecten maximus and Mytilus galloprovincialis (Pteriomorphia), and Yoldia eightsi (Protobranchia). Comparison with the Hox and ParaHox genes of other bilaterians, particularly lophotrochozoans, allowed us to attribute six of these sequences to the Hox gene cluster (one to paralog group [PG] 3 class, and five to the central class), two to the ParaHox cluster and one to the Gbx gene family.The results of our investigation seem to indicate that homeotic Hox and ParaHox gene clusters are homogeneous for both presence and characteristics in molluscs.     

Monoaminergic and cholinergic mechanisms of reproduction control in marine bivalve molluscs and echinoderms: A review

The autoreproduction ability is one of the most important properties of living systems. Evolution of metazoans ensured their reproduction by means of such determinants as sexual cells developing in specialized organs, in gonads. In most marine invertebrates, the gonad produces tens of millions of gametes per reproductive cycle. This reproduction level guarantees the species resistance in the external environment and is provided by the whole organism, though in unfavourable ecological conditions the gonad may become a source of trophic and energy material and it can maintain the viability of organism by means or the mass, often total, lysis of sexual cells. This metabolic interaction of a part (gonad) and the whole (organism) presumes the existence of the strictly determined relations between them, on the one hand, and more or less pronounced autonomy, on the other hand. The isolated organs, including gonads, are capable of fulfiling specific functions for a short period of time by means of local regulation. However, there is no full autonomy between the gonad and the organism: the hierarchical relations set in between them.     

Anaerobic metabolism of bivalve molluscs during exposure to air

The anaerobic metabolism of Mytilus edulis, Cardium edule, Scrobicularia plana and Macoma balthica was investigated. On exposure to the atmosphere, all of these species were found to be able to utilise ¹⁴CO₂. This suggests that these species gape during exposure to the atmosphere. A comparative study on the pattern of ¹⁴C incorporation suggests that there is a similarity between M. edulis and S. plana in the extent of the utilization of the succinate pathway during exposure to air. However C. edule and M. balthica were also similar in the extent of utilizing the succinate pathway, even though there were significant species differences between the similar animals. It is suggested that exposure in S. plana represents a stressful situation and that this species might react to this stress by utilizing the succinate pathway. A higher incorporation of radiocarbon into alanine by M. balthica could be due to high activity of the enzymes that control the reactions leading to production of radioactive pyruvate.     

Analysis of different methods to estimate fecundity in bivalve molluscs

Abstract

We analyzed the advantages and disadvantages of three methods to estimate potential fecundity in the bivalve molluscs Pinctada mazatlanica and Atrina maura, both species having commercial value in northwestern Mexico. Data of gonad samples collected during the breeding season in March 1999 (P. mazatlanica) and March 2003 (A. maura) were processed with histological techniques combined with Cavalieri’s Principle; stereological analysis of the gonad with the caliper method; and estimation of the theoretical radius of oocytes. These estimates were compared to a reference value obtained from direct counts of the number of oocytes stripped from the gonad. Compared to other methods, potential fecundity determined with the caliper method was more accurate in both species: A. maura (9.8–15 × 10⁶ cells ind^?1 and P. mazatlanica (10.8–17 × 10⁶ cells ind^?1). The potential of the caliper method, combined with image analysis software may offer accurate estimates of aspects of reproduction in different bivalve species, which has direct applications in hatcheries and conservation programs.     

Effect of elevated temperature and of hypoxia on NO activity in the central nervous system of bivalve molluscs

By light and electron microscopy methods the effect of changes of environmental conditions on the state of the nitroxidergic system has been studied in molluscs on the background of action of elevated temperature and hypoxia. Analysis is performed of biological effect of isolated and combined effects of the studied factors on dynamics of NO synthesis. A higher resistance of CNS neurons to the combined action of hyperthermia and hypoxia is revealed in molluscs with the initially high level of nitrogen oxide production. In molluscs with the initially low level of development of the nitroxidergic system, induction of NO formation in stress has been found to be accompanied by a change of morphology of nervous structures. It is suggested that nitrogen oxide participates in evolutionary established mechanisms of protection of mollusc nerve cells from hypoxia, while the initial high level of NO production reflects larger adaptational possibilities characteristic of these organisms.     

Monoaminergic and cholinergic mechanisms of reproduction control in marine bivalve molluscs and echinoderms: a review.

The autoreproduction ability is one of the most important properties of living systems. Evolution of metazoans ensured their reproduction by means of such determinants as sexual cells developing in specialized organs, in gonads. In most marine invertebrates, the gonad produces tens of millions of gametes per reproductive cycle. This reproduction level guarantees the species resistance in the external environment and is provided by the whole organism, though in unfavourable ecological conditions the gonad may become a source of trophic and energy material and it can maintain the viability of organism by means or the mass, often total, lysis of sexual cells. This metabolic interaction of a part (gonad) and the whole (organism) presumes the existence of the strictly determined relations between them, on the one hand, and more or less pronounced autonomy, on the other hand. The isolated organs, including gonads, are capable of fulfilling specific functions for a short period of time by means of local regulation. However, there is no full autonomy between the gonad and the organism: the hierarchical relations set in between them.     

Adaptations to hypoxia in hydrothermal-vent and cold-seep invertebrates

The deep sea harbors very unusual environments, such as hydrothermal vents and cold seeps, that illustrate an apparent paradox: the environmental conditions are very challenging and yet they display a high biomass when compared to the surrounding environment at similar depth. Hypoxia is one of the challenges that these species face to live there. Here, we review specific adaptations of their respiratory system that the species have developed to cope with hypoxia, at the morphological, physiological, and biochemical levels. Most studies to date deal with annelids and crustaceans, and trends can be drawn: development of ventilation and branchial surfaces to help with oxygen extraction, and an increase in finely tuned oxygen binding proteins to help with oxygen storage and transport. Beside these respiratory adaptations most animals have developed enhanced anaerobic capacities and specific ways to deal with sulfide.     

Calibration of the impedance method for rapid quantitative estimation of Escherichia coli in live marine bivalve molluscs

AIM: Calibration of impedance measurement was performed vs the Association Fran?oise de Normalisation (AFNOR) MPN method with a view to rapid enumeration of Escherichia coli in live marine bivalve molluscs. METHODS AND RESULTS: Linear regression models between log10 MPN and detection time (DT) were adjusted for several shellfish types, growth media, and impedance instruments (BacTrac and Malthus systems). Escherichia coli concentrations could be estimated from DT using a single regression line for BacTrac 4100 with M1 medium (R2 = 87.8%) and Malthus with M2 medium (R2 = 86.7%), and two regression lines for BacTrac 4110 with M2 medium (R2 = 86.4 and 88.2%). The uncertainty of the predicted bacterial concentration was around +/-0.43 log unit for duplicate sample analysis. The impedance signal was attributable to E. coli in 99% of cases. All cultures containing E. coli produced an impedance signal with BacTrac 4100 and BacTrac 4110, whereas 5.6% did not exhibit a signal with Malthus. CONCLUSIONS: Impedance measurement is a possible alternative to the MPN method for rapid quantitative estimation of E. coli in live bivalve shellfish. SIGNIFICANCE AND IMPACT OF THE STUDY: The impedance method reduces analysis handling time considerably and is much easier to use than the MPN method. Moreover, results can be obtained within 5-10 h, allowing rapid intervention to ensure public health protection in case of shellfish contamination.     

The ecology of Bonamia and decline of bivalve molluscs

Bonamia is a protozoan parasite of the haemocytes of oysters (Tiostrea chilensis), in which it has an annual developmental cycle between November and August each year. The parasite transmits directly, oyster to oyster, and therefore disease spread is related to host stock density. The Foveaux Strait oyster population experiences large mortalities every 20-30 years, and these may be attributable to Bonamia. The parasite appears to become less pathogenic at the end of, and probably between, mass mortalities, and some oysters appear more tolerant of infection than others. On the basis of these observations, and considering other protist pathogen:oyster models, the apparently reduced pathogenicity of Bonamia is discussed in terms of parasite kinetics. The population dynamics and selection of parasite tolerant host stocks, and kinetics of parasite transmission, may explain the cyclic nature of large-scale mortalities in Foveaux Strait, without change in parasite pathogenicity.