Species Crassostrea Gigas Research Articles

Variation and characterization of prometryn in oysters (Crassostrea gigas) after seawater exposure

Prometryn (PRO) is frequently detected in shellfish of international trade among triazine herbicides because of its wide application in agriculture and aquaculture worldwide. Nevertheless, the variations of PRO remain unclear in aquatic organisms, which affect the accuracy of their food safety risk assessment. In the present study, the tissue-specific accumulation, biotransformation, and potential metabolic pathway of PRO were reported in oyster species Crassostrea gigas for the first time. The experiments were conducted through semi-static seawater exposure with low and high concentrations of PRO (at nominal concentrations of 10 and 100 μg/L) via daily renewal over 22 days, followed by 16 days of depuration in clean seawater. The characterization of prometryn in oysters was then evaluated through the bioaccumulation behavior, elimination pathway and metabolic transformation, comparing with other organisms. The digestive gland and gonad were found to be the main target organs during uptake. In addition, the highest bioconcentration factor of 67.4 ± 4.1 was observed when exposed to low concentration. The level of PRO in oyster tissues rapidly decreased within 1 day during depuration, with an elimination rate of >90 % for the gill. Moreover, four metabolites of PRO were identified in oyster samples of exposed groups, including HP, DDIHP, DIP, and DIHP, in which HP was the major metabolite. Considering the mass percentage of hydroxylated metabolites higher than 90 % in oyster samples, PRO poses a larger threat to aquatic organisms than rat. Finally, the biotransformation pathway of PRO in C. gigas was proposed, the major metabolic process of which was hydroxylation along with N-dealkylation. Meanwhile, the newly discovered biotransformation of PRO in oyster indicates the importance of monitoring environmental levels of PRO in cultured shellfish, to prevent possible ecotoxicological effects as well as to ensure the safety of aquatic products.

Analysis of Lead (Pb) and Cadmium (Cd) in Oyster Crassostrea gigas and Saccostrea cucullata using Atomic Absorption and Ultraviolet-Visible Spectrophotometer Methods

Lead (Pb) and Cadmium (Cd) metals are highly toxic when accumulated in the body and cause serious diseases and even death. This study aimed to determine the metal content of Pb and Cd in the oyster species Crassostrea gigas and Saccostrea cucullate in the Lampulo and Krueng Cut areas at three different places. Analysis of the metals was carried out using AAS and UV-Visible spectrophotometers. Samples were prepared using concentrated HNO3 wet destruction and the addition of dithizone. The results showed Pb measured by AAS in Saccostrea cucullata 2.33-8.00 ppb, and by UV-Vis 2.77-8.66 ppb. Pb within Crassostrea gigas by AAS method 1.77-10.30 ppb, and by UV-Vis method 2.11-10.66 ppb. The Cd metal measured by AAS in Saccostrea cucullate is 3.80-10.50 ppb, and by UV-Vis method is 4.29-10.77 ppb, whereas Cd in Crassostrea gigas measured by AAS is 12.11-13.22 ppb and by UV-Vis method is 12.77-13.66 ppb. Linearity of Pb measurements with AAS and UV-Vis was obtained R2 =0.9979 and R2 =0.9938 and for Cd were R2 =0.9986 and R2 =0.9810. Accuracy (%recorvery) of Pb measurements by AAS and UV-Vis are 80-110% and 98-113% whereas Cd showed 100-106% and 91-107% respectively. The relative value of the standard deviation (%RSD ± SD) is 0.001 indicating excellent measurement results. Based on t-test calculations, measurements of Pb and Cd levels showed no difference between the AAS method and the UV-Vis method

A sustainable one-pot method to transform seashell waste calcium carbonate to osteoinductive hydroxyapatite micro-nanoparticles.

We have developed a straightforward, one-pot, low-temperature hydrothermal method to transform oyster shell waste particles (bCCP) from the species Crassostrea gigas (Mg-calcite, 5 wt% Mg) into hydroxyapatite (HA) micro/nanoparticles. The influence of the P reagents (H3PO4, KH2PO4, and K2HPO4), P/bCCP molar ratios (0.24, 0.6, and 0.96), digestion temperatures (25-200 °C), and digestion times (1 week-2 months) on the transformation process was thoroughly investigated. At 1 week, the minimum temperature to yield the full transformation significantly reduced from 160 °C to 120 °C when using K2HPO4 instead of KH2PO4 at a P/bCCP ratio of 0.6, and even to 80 °C at a P/bCCP ratio of 0.96. The transformation took place via a dissolution-reprecipitation mechanism driven by the favorable balance between HA precipitation and bCCP dissolution, due to the lower solubility product of HA than that of calcite at any of the tested temperatures. Both the bCCP and the derived HA particles were cytocompatible for MG-63 human osteosarcoma cells and m17.ASC murine mesenchymal stem cells, and additionally, they promoted the osteogenic differentiation of m17.ASC, especially the HA particles. Because of their physicochemical features and biological compatibility, both particles could be useful osteoinductive platforms for translational applications in bone tissue engineering.

Tropical and Subtropical Ostreidae of the American Pacific: Fisheries, Aquaculture, Management, and Conservation

Oysters are one of the most important groups in terms of production by fisheries and aquaculture in developed countries of temperate areas. In subtropical and tropical regions, oyster species are economic important resources but many are exploited in an artisanal way and without a legal framework, what can result in overexploitation. One alternative is aquaculture but biological information and development of techniques for local species are required. Therefore, this review organizes information on subtropical and tropical oysters from the American Pacific addressing on their fisheries, cultivation, management, and conservation. The study focuses on two native species Striostrea prismatica and Crassostrea corteziensis and two introduced species Crassostrea gigas and Crassostrea sikamea having more information and technological development. Fisheries and their problems are analyzed and some alternatives are proposed to improve their organization and preservation. The most important aquaculture technological aspects are described including the processes for the production of spat in hatcheries and the techniques used for field cultivation. Biological, physiological, and ecological studies as well as the implementation of legal measures are required to administrate, restore, and recover fisheries. The aquaculture of C. corteziensis and S. prismatica presents great advances but the other native species require the development of technologies to progress in many aspects. The production of C. sikamea is concentrated in one region without expecting further expansion, and the cultivation of C. gigas can be recreated on various sites and expanded to estuaries in tropical areas.

Amazonia Seasons Have an Influence in the Composition of Bacterial Gut Microbiota of Mangrove Oysters (Crassostrea gasar).

The mangrove oysters (Crassostrea gasar) are molluscs native to the Amazonia region and their exploration and farming has increased considerably in recent years. These animals are farmed on beds built in the rivers of the Amazonia estuaries and, therefore, the composition of their microbiome should be directly influenced by environmental conditions. Our work aimed to evaluate the changes in bacterial composition of oyster's microbiota at two different seasons (rainy and dry). For this purpose, we amplified and sequenced the V3-V4 regions of the 16S rRNA gene. Sequencing was performed on the Illumina MiSeq platform. According to the rarefaction curve, the sampling effort was sufficient to describe the bacterial diversity in the samples. Alpha-diversity indexes showed that the bacterial microbiota of oysters is richer during the rainy season. This richness is possibly associated with the diversity at lower taxonomic levels, since the relative abundance of bacterial phyla in the two seasons remained relatively constant. The main phyla found include Firmicutes, Bacteroidetes, Actinobacteria, and Proteobacteria. Similar results were found for the species Crassostrea gigas, Crassostrea sikamea, and Crassostrea corteziensis. Beta-diversity analysis showed that the bacterial composition of oyster's gut microbiota was quite different in the two seasons. Our data demonstrate the close relationship between the environment and the microbiome of these molluscs, reinforcing the need for conservation and sustainable management of estuaries in the Amazonia.

Twists and turns of an oyster's life: effects of different depuration periods on physiological biochemical functions of oysters.

Aquaculture activities are often established in the vicinity of highly populated, potentially contaminated areas. Animals cultured at such locations, namely bivalves, are frequently used as test organisms in ecotoxicological testing. In this case, a period of depuration is required to allow the normalization of physiological processes, which are likely to be altered after exposure to a multiplicity of waterborne contaminants occurring in the wild. One of the most important species in modern marine aquaculture is the oyster species Crassostrea gigas. The aim of this study was to assess if the current depuration time frame of 24 h (adopted by most aquaculture facilities), is long enough to permit oysters to revert potential toxic effects exerted by environmental contaminants, allowing their use in laboratory-based ecotoxicological studies. The selected approach involved the monitoring of biochemical (antioxidant defence, oxidative damage, phase II metabolism, and neurological homeostasis) and physiological (condition index) parameters, along a period of 42 days. The obtained results showed that a period of 24 h does not revert any of the potential toxic effects caused by environmental contaminants to which animals may have been previously subjected; even a period of 42 days was not long enough for the oysters to completely normalize the levels of their antioxidant defences, namely total GPx activity, which increased over time. Lipid peroxidation was also increased during the depuration period, and the activity of the metabolic isoenzymes GSTs was significantly decreased. Furthermore, AChE activity measured in the adductor muscle of oysters was increased over time. These assumptions suggest that a period of depuration longer than 24 h is mandatory to obtain adequate test organisms of this oyster species, to be used for ecotoxicological testing purposes.

First Report of an Eosinophilic Rickettsia-like Organism in Diseased Oysters Crassostrea gigas

ABSTRACT Eosinophilic Rickettsia-like organisms (E-RLO) were found for the first time in sick oysters of the species Crassostrea gigas, grown in Laguna de San Ignacio, Baja California Sur, Mexico, in August 2014. This work presents the histological description of development of bacteria in parasitophorous vacuoles of infected cells. Confirmative diagnosis was performed by polymerase chain reaction and in situ hybridization. Bacterial growth started with eosinophilic granules (colonial), followed by the formation and growth of intracytoplasmic inclusions. The presence of E-RLO was detected in the gill epithelium, mantle, labial palps, digestive tract, and both male and female gametes. This work describes for the first time an E-RLO in C. gigas with potential vertical transmission.

The paralytic shellfish toxin, saxitoxin, enters the cytoplasm and induces apoptosis of oyster immune cells through a caspase-dependent pathway

Exposure of the toxin-producing dinoflagellate Alexandrium catenella (A. catenella) was previously demonstrated to cause apoptosis of hemocytes in the oyster species Crassostrea gigas. In this work, a coumarin-labeled saxitoxin appeared to spread throughout the cytoplasm of the hemocytes. PSTs, including saxitoxin, were also shown to be directly responsible for inducing apoptosis in hemocytes, a process dependent on caspase activation and independent of reactive oxygen species (ROS) production. A series of in vitro labelling and microscopy experiments revealed that STX and analogs there of induced nuclear condensation, phosphatidylserine exposure, membrane permeability, and DNA fragmentation of hemocytes. Unlike in vertebrates, gonyautoxin-5 (GTX5), which is present in high concentrations in A. catenella, was found to be more toxic than saxitoxin (STX) to oyster immune cells. Altogether, results show that PSTs produced by toxic dinoflagellates enter the cytoplasm and induce apoptosis of oyster immune cells through a caspase-dependent pathway. Because of the central role of hemocytes in mollusc immune defense, PST-induced death of hemocytes could negatively affect resistance of bivalve molluscs to microbial infection.

High-Resolution Melting of 18S rDNA sequences (18S-HRM) for discrimination of bivalve’s species at early juvenile stage: application to a spat survey

It is particularly difficult to identify bivalve species at early development stages (larvae and spat) by morphological examination. Targeting genetic marker commonly used for species identification, we developed a cheap and user friendly tool for bivalve identification through High-Resolution Melting (HRM) analysis. Based on the polymorphism of the nuclear 18S ribosomal RNA gene among 11 orders of bivalve, the 18S-HRM method allowed discriminating species of the Ostreida, Mytilida and Pectinida orders. In this study, this method proved to be a very sensitive tool able to authenticate one species that differed by only one mutation from another one. We applied the method by identifying bivalve species in spat collections from different benthic substrates moored at subtidal and intertidal areas in euhaline conditions. This field survey showed that the settlement of the invasive species Crassostrea gigas was limited vertically to intertidal substrates. In conclusion, the 18S-HRM analysis is a fast, closed-tube method with high resolution and cost effectiveness, and is thus a good alternative method to identify bivalve species. This HRM assay can by applied for species identification of bivalves in the context of ecological management as well as food traceability for bivalves with economical interests.

Sperm Accumulated Against Surface: A novel alternative bioassay for environmental monitoring

Forecasting the impacts of changes in water quality on broadcast spawning aquatic organisms is a key aspect of environmental monitoring. Rapid assays of reproductive potential are central to this monitoring, and there is a need to develop a variety of methods to identify responses. Here, we report a proof-of-concept study that assesses whether quantification of “Sperm Accumulated Against Surface” (SAAS) of tissue culture well-plates could be a rapid and simple proxy measure of fertilisation success. Our results confirm that motile sperm (but not immotile sperm) actively accumulate at surfaces and that the pattern of accumulation reflects fertilisation success in the model oyster species Crassostrea gigas. Furthermore, we confirm these patterns of SAAS for another marine species, the polychaete Galeolaria caespitosa, as well as for a freshwater species, the fish Gasterosteus aculeatus. For all species considered, SAAS reflected changes in sperm performance caused by experimentally manipulated differences in water quality (here, salinity). These findings indicate that SAAS could be applied easily to a range of species when examining the effects of water quality. Measurement of SAAS could, therefore, form the basis of a rapid and reliable assay for bioassessments of broadcast spawning aquatic organisms.

Revision of genusCrassostrea(Bivalvia: Ostreidae) of Brazil

The Ostreidae are well known for their high intra-specific variation, which makes identification problematic. The present paper aims to provide a morpho-anatomical and taxonomic review of the Brazilian species ofCrassostrea, as well as some congeneric species from other relevant areas. The Brazilian species areCrassostrea manglesp.nov. andCrassostrea brasiliana. The exotic speciesCrassostrea gigas, cultivated in the southern region of the country, is also included in this study. Additional species are:Crassostrea virginica, from the Atlantic coast of the USA, the type species, andCrassostrea rhizophorae, a south-eastern Caribbean species that is supposed to occur in Brazil, but is here understood as a different entity endemic to that region. Taking into account their economic importance, the differentiation between these species is critical, and is essential for a better planning of production and preservation strategies. We point out the differences in almost all structures amongst these species, which are formally redescribed herein. The occurrence of the African speciesCrassostrea gasarin Brazil is established as false. The voucher specimens of previous studies responsible for this assumption were examined, evidencing a misidentification forCrassostrea brasilianafrom an estuarine environment. The geographical distribution of the studied species is recognized as follows:Crassostrea rhizophoraein the south-east Caribbean Sea;Crassostrea manglesp. nov. from Pará to the Santa Catarina, only in mangroves;Crassostrea brasilianafrom Paraíba to Santa Catarina, both in rocky shores and mangroves;Crassostrea virginicaranges from the Atlantic coast of Canada to the Caribbean; andCrassostrea gigasoriginates from the Indo-Pacific, but has been introduced in southern Brazil. This paper also deals with conchological aspects of the endemic speciesCrassostrea praia, from south Lagoa dos Patos, Rio Grande do Sul; after a more detailed definition, and considering the deleterious effects of the nearby port and construction sites, it can be classified as an endangered species.

Comparison of haemocytic parameters among flat oyster Ostrea edulis stocks with different susceptibility to bonamiosis and the Pacific oyster Crassostrea gigas

Farming of the flat oyster Ostrea edulis in Europe is severely constrained by the protozoan Bonamia ostreae. The introduction of the resistant species Crassostrea gigas has been a relief for the farmers, while the pilot programmes to select O. edulis strains resistant to bonamiosis performed in various countries can be seen as a promising strategy to minimise the effects of bonamiosis. However, the physiological bases of this differential susceptibility remain unknown. A search for an explanation of the intra and interspecific differences in oyster susceptibility to bonamiosis was accomplished by comparing some immune parameters among various O. edulis stocks and C. gigas. On December 2003, naïve and Bonamia-relatively resistant flat oysters from Ireland, Galician flat oysters and Pacific oysters C. gigas were deployed in a Galician area affected by bonamiosis; haemolymph samples were taken in February and May 2004. A new oyster deployment at the same place was carried out on June 2004 and haemolymph sampling was performed on April 2005. On November 2004, new sets of Irish flat oysters and C. gigas were deployed in Ireland and haemolymph sampling was performed in June 2005. Various haemocytic parameters were measured: total and differential haemocyte count, phagocytic ability, respiratory burst (superoxide anion [O2-] and hydrogen peroxide [H2O2]) and nitric oxide [NO] production. The comparison of the parameters was carried out at 3 levels: (1) between O. edulis and C. gigas, (2) among O. edulis stocks with different susceptibility to bonamiosis, and (3) between Bonamia-infected and non infected O. edulis. In addition, haemocyte-B. ostreaein vitro encounters were performed to analyse interspecific differences in the haemocytic respiratory burst, using flow cytometry. Significant differences associated with total and differential haemocyte count, and respiratory burst between O. edulis and C. gigas were detected, which could be linked to differences in susceptibility to bonamiosis between both species. Additionally, significant changes in total and differential haemocyte count, and respiratory burst of O. edulis associated with B. ostreae infection were found. However, no consistent difference in any haemocyte parameter between the O. edulis stocks involved in the study was recorded.

Biological record of added manganese in seawater: a new efficient tool to mark in vivo growth lines in the oyster species Crassostrea gigas

The biological response of increased manganese in seawater was tested experimentally with the oyster species Crassostrea gigas by adding, once per day, a fixed quantity of MnCl2 to the container where the oysters were living. Uptake of Mn2+ in the shell was traced with cathodoluminescence and quantified with a high spatial resolution proton microprobe. The daily addition of MnCl2 resulted in the visualization of distinct growth increments seen simultaneously in both the calcitic shell and the aragonitic ligament. A relation was observed between the addition of Mn2+ to the seawater and incorporation of Mn in the mineral part of the shell. Thus, addition of MnCl2 to seawater is an efficient tool to mark in vivo growth increments in bio-mineralised carbonates.

Incorporation and modification of dietary fatty acids in gill polar lipids by two bivalve species Crassostrea gigas and Ruditapes philippinarum

Two bivalve species Crassostrea gigas and Ruditapes philippinarum were fed eight weeks with three mono-specific algae diets: T- Isochrysis galbana, Tetraselmis suecica, Chaetoceros calcitrans, selected on the basis of their polyunsaturated fatty acid (PUFA) composition. The incorporation and the modification of dietary fatty acids in C. gigas and R. philippinarum gill lipids were analysed and compared. Essential PUFA (20:4n-6, 20:5n-3 and 22:6n-3) and non-methylene interrupted PUFAs (known to be synthesised from monounsaturated precursors) contents of gill polar lipid of both species were greatly influenced by the dietary conditioning. Interestingly, oysters and clams responded differentially to the mono-specific diets. Oysters maintained higher 20:5n-3 level and higher 22:2j/22:i and n-7/n-9 ratio in gill polar lipids than clams. To better discriminate dietary and species influences on the fatty acid composition, a Principal Component Analysis followed by a MANOVA on the two most explicative components was performed. These statistical analyses showed that difference in fatty acid compositions attributable to species were just as significant as the diet inputs. The differences of gill fatty acid compositions between oysters and clams are speculated to result of an intrinsic species characteristic and perhaps of a group characteristic: Fillibranch vs. Eulamellibranch.

Elements of reproductive strategy in oysters, Crassostrea gigas, from the “Rade de Brest”, France

Summary In France oyster farming is very important from an economical point of view (150,000 t/y), but curiously, no data exist concerning the reproductive cycle of the cultivated species Crassostrea gigas. New selection programs in oyster hatcheries are based on the use of adults having proper characteristics (growth, resistance to diseases and known sexuality) for which we need to know how sex determination can be influenced by environmental factors. C. gigas individuals from a natural population of the “Rade de Brest”, Brittany, France, were examined every 2 weeks during a 2-year period from February 1996. Reproductive parameters including their condition index and their sex ratio were determined, and seasonal and annual differences assessed. Under microscopic examination the presence of sperm or oocytes or the absence of gametes allowed us to define individuals as male, female, hermaphrodite or sexually undetermined oysters. The maximum percentage of female oysters, 70%, was found in July, whereas males were observed almost throughout the year. Females exhibited a very distinct seasonal pattern of gametogenesis. The frequency of hermaphrodites varied from less than 1% during July—September to about 37% during May—April. The sex ratio was three females to one male just before the spawning season. In conclusion, sex determination of C. gigas oysters is strongly influenced by environmental factors. Therefore, further study of the influence of such factors is of great interest for practical applications in the hatchery such as the sex determination of breeders.

Present knowledge on the molecular basis of cytotoxicity, antibacterial activity and stress response in marine bivalves

Abstract Mussel hemolymph possessed naturally occurring cytotoxic activity. Vertebrate erythrocytes, mouse tumor cell line and parasite Bonamia were killed, but not bacteria. Two step chromatography revealed a complex multimeric protein of 320 kDa, acting through a polymerization process on the target cell membrane. Of naive animals from the two oyster species Crassostrea gigas and Ostrea eclulis, a low proportion of plasmas and hemocyte lysates presented naturally occurring antibacterial activity. Induction assays by injection of saline solution or bacterial suspensions resulted in increased activity in virtually all oysters. Identification of active polypeptides included acidic solubility followed by reverse phase chroma‐tographies. Oyster hemocytes responded to heat‐shock by the synthesis of particular proteins. Treated in the same way, the protozoan parasite Perkinsus marinus also synthesised long‐lasting 70 to 30 kDa proteins which are different from the oyster ones. The question arises of the biolog...