Crassostrea Species Research Articles

Comparison of the effects of reef and anthropogenic soundscapes on oyster larvae settlement

Settlement is a critical period in the life cycle of marine invertebrates with a planktonic larval stage. For reef-building invertebrates such as oysters and corals, settlement rates are predictive for long-term reef survival. Increasing evidence suggests that marine invertebrates use information from ocean soundscapes to inform settlement decisions. Sessile marine invertebrates with a planktonic stage are particularly reliant on environmental cues to direct them to ideal habitats. As gregarious settlers, oysters prefer to settle amongst members of the same species. It has been hypothesized that oyster larvae from species Crassostrea virginica and Ostrea angasi use distinct conspecific oyster reef sounds to navigate to ideal habitats. In controlled laboratory experiments we exposed Pacific Oyster Magallana gigas larvae to anthropogenic sounds from conspecific oyster reefs, vessels, combined reef-vessel sounds as well as off-reef and no speaker controls. Our findings show that sounds recorded at conspecific reefs induced higher percentages of settlement by about 1.44 and 1.64 times compared to off-reef and no speaker controls, respectively. In contrast, the settlement increase compared to the no speaker control was non-significant for vessel sounds (1.21 fold), combined reef-vessel sounds (1.30 fold), and off-reef sounds (1.18 fold). This study serves as a foundational stepping stone for exploring larval sound feature preferences within this species.

Genetic evidence of the unique identity of the West African Mangrove Oyster (Crassostrea tulipa) from the Gulf of Guinea

This research enabled the genetic identification of the West African mangrove oysters, Crassostrea tulipa, as well as establishing the evolutionary relationship between it and other Crassostrea species. Essentially, the study assisted in clearing up a long-standing confusion over this species’ synonymy with C. gasar. Also, the population structure of 60 C. tulipa individuals, from three different ecotypes, was analyzed using the mitochondrial cytochrome oxidase I (COI) genes as a marker. Results provided the first genetic sequences for C. tulipa and deposited in the GeneBank. Optimal and consensus bootstrap Neighbor-Joining trees distinctively differentiated C. tulipa from other Crassostrea species and consistently formed a different clade with C. gasar, with no bootstrap value from either NJ, MPT, or UPG trees supporting their similarity. C. tulipa sequences occurred as different haplotypes from other Crassostrea sp, with a mutation value as high as 288 and a haplotype diversity of 0.893 between C. tulipa and C. gasar sequences. High estimates of genetic distance (1.40–1.55) and patristic divergence were recorded between C. tulipa and C. gasar, in the same range as with seven other species. The study thus reveals the unique identity of C. tulipa as genetically distinct from C. gasar and other Crassostrea species. Based on the population structure analyses from the neutrality test, a low to high haplotype diversity h (0.000–0.963) and low nucleotide diversity π (0.00–0.378) were obtained. A negative mean Tajima’s D (−0.65247), and a positive Fu’s Fs (3.194), suggest rare variations or low-frequency polymorphisms. In addition to serving as the basis for phylogeny, the identification of C. tulipa and the recent data on its population structure also serve as the basis for conservation efforts and hatchery-based aquaculture.

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 historical and integrative taxonomic account of mangrove oyster species native to the Atlantic American coast: A re-evaluation of Brazilian Crassostrea species

The identification of native mangrove oyster species of the genus Crassostrea in Brazil is a recurring issue for specialists. Studies have recognized the presence of two species in the littoral zone: C. rhizophorae and C. gasar. However, other names may be used, such as C. mangle for C. rhizophorae and C. brasiliana or C. tulipa for C.gasar. As in most oyster taxonomic studies, their identification requires genetic evaluation for the precise determination of taxa. Moreover, previous revisions have proposed some morphological traits by which each species could be precisely identified, leading to a series of debates about the best model for species identity. The aim of this study was to apply an integrative taxonomic approach to native Crassostrea mangrove oyster species on the Brazilian coast to assert the correct species names, delimitations, and concise descriptions. Molecular tracing of cultivated specimens, along with wild specimens of Crassostrea collected from the Brazilian coast, was used to confirm the morphological identity of the taxa. The wild specimens were tentatively identified as either C. gasar or C. rhizophorae based on previous morphological revisions. PCR-RFLP and DNA analyses of the mitochondrial 16S gene mitochondrial rDNA from wild specimens were used to confirm taxa and re-evaluate morphological boundaries and descriptions of the species. We confirmed two patterns in the RFLP and sequence analyses: C. gasar and C. rhizophorae. Conchological and anatomical evaluations of 1006 identified Crassostrea specimens confirmed the inherent polymorphism between the native species, preventing reliable identification of the species by morphology alone. A clear exception, however, is found for specimens whose shell height is greater than 110 mm with internal view of white-colored valves and without an umbonal cavity, fitting the description of C. paraibanensis, which belongs to C. gasar. C. brasiliana and C. mangle are synonyms of C. gasar and C. rhizophorae, respectively. C. tulipa represents a species disconnected from any described mangrove oyster species from Atlantic seas. Conchological and anatomical descriptions of both species were improved to include the variability encountered.

Comparative biochemical and molecular responses of biotransformation and antioxidant systems in three species of Crassostrea (Sacco, 1897) oysters exposed to chrysene

Chrysene (CHR) is among the most persistent polycyclic aromatic hydrocarbons (PAH) in water and a priority compound for pollutants monitoring, due to its carcinogenic, mutagenic and genotoxic potential. Aquatic animals exposed to CHR may present alterations of biomarkers involved in the biotransformation and oxidative stress-related parameters. The aim of this study was to investigate differences in antioxidant and biotransformation (phase I and II) systems of Crassostrea gigas, C. gasar and C. rhizophorae and its effects resulting from CHR exposure. Adult oysters of these species were exposed to 10 μg L−1 of CHR for 24 h and 96 h. In gills, the transcripts CYP1-like, CYP2-like, CYP2AU1-like, GSTO-like, MGST-like, SULT-like were evaluated after 24 h of exposure. The activity of SOD, CAT, GPx, GR and G6PDH were analyzed in gills and digestive glands after 96 h of exposure. CHR bioaccumulated in tissues. Differences in the remaining levels of CHR in water after 96 h were observed in aquaria containing C. gigas or C. gasar oysters and may be associated to the different filtration rates between these species. Downregulate of biotransformation genes were observed in gills of C. gasar (CYP2AU1-like and GSTO-like) and C. rhizophorae (CYP1-like1, CYP2-like, MGST-like and SULT-like), suggesting that biotransformation responses may be species-specific. Differential activity of antioxidant enzymes were observed in gills and digestive gland of oysters exposed to CHR. Biochemical responses suggested that C. gigas and C. gasar are more responsive to CHR. Differential responses observed among the three Crassostrea species can be related to evolutionary differences, ecological niches and adaptation to environment.

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.

Construction and analysis of the chromosome-level haplotype-resolved genomes of two Crassostrea oyster congeners: Crassostrea angulata and Crassostrea gigas.

The Portuguese oyster Crassostrea angulata and the Pacific oyster C. gigas are two major Crassostrea species that are naturally distributed along the Northwest Pacific coast and possess great ecological and economic value. Here, we report the construction and comparative analysis of the chromosome-level haplotype-resolved genomes of the two oyster congeners. Based on a trio-binning strategy, the PacBio high-fidelity and Illumina Hi-C reads of the offspring of the hybrid cross C. angulata (♂) × C. gigas (♀) were partitioned and independently assembled to construct two chromosome-level fully phased genomes. The assembly size (contig N50 size, BUSCO completeness) of the two genomes were 582.4M (12.8M, 99.1%) and 606.4M (5.46M, 98.9%) for C. angulata and C. gigas, respectively, ranking at the top of mollusk genomes with high contiguity and integrity. The general features of the two genomes were highly similar, and 15,475 highly conserved ortholog gene pairs shared identical gene structures and similar genomic locations. Highly similar sequences can be primarily identified in the coding regions, whereas most noncoding regions and introns of genes in the same ortholog group contain substantial small genomic and/or structural variations. Based on population resequencing analysis, a total of 2,756 species-specific single-nucleotide polymorphisms and 1,088 genes possibly under selection were identified. This is the first report of trio-binned fully phased chromosome-level genomes in marine invertebrates. The study provides fundamental resources for the research on mollusk genetics, comparative genomics, and molecular evolution.

Wild oyster population resistance to ocean acidification adversely affected by bacterial infection

The carbon dioxide induced ocean acidification (OA) process is well known to have profound effects on physiology, survival and immune responses in marine organisms, and particularly calcifiers including edible oysters. At the same time, some wild populations could develop a complex and sophisticated immune system to cope with multiple biotic and abiotic stresses, such as bacterial infections and OA, over the long period of coevolution with the environment. However, it is unclear how immunological responses and the underlying mechanisms are altered under the combined effect of OA and bacterial infection, especially in the ecologically and economically important edible oysters. Here, we collected the wild population of oyster species Crassostrea hongkongensis (the Hong Kong oyster) from their native estuarine area and carried out a bacterial challenge with the worldwide pervasive pathogen of human foodborne disease, Vibrio parahaemolyticus, to investigate the host immune responses and molecular mechanisms under the high-CO2 and low pH-driven OA conditions. The wild population had a high immune resistance to OA, but the resistance is compromised under the combined effect of OA and bacterial infection both in vivo or in vitro. We classified all transcriptomic genes based on expression profiles and functional pathways and identified the specifically switched on and off genes and pathways under combined effect. These genes and pathways were mainly involved in multiple immunological processes including pathogen recognition, immune signal transduction and effectors. This work would help understand how the immunological function and mechanism response to bacterial infection in wild populations and predict the dynamic distribution of human health-related pathogens to reduce the risk of foodborne disease under the future climate change scenario.

Epigenetic-associated phenotypic plasticity of the ocean acidification-acclimated edible oyster in the mariculture environment.

For marine invertebrates with a pelagic-benthic life cycle, larval exposure to ocean acidification (OA) can affect adult performance in response to another environmental stressor. This carry-over effect has the potential to alter phenotypic traits. However, the molecular mechanisms that mediate "OA"-triggered carry-over effects have not been explored despite such information being key to improving species fitness and management strategies for aquafarming. This study integrated the genome-wide DNA methylome and transcriptome to examine epigenetic modification-mediated carry-over OA impacts on phenotypic traits of the ecologically and commercially important oyster species Crassostrea hongkongensis under field conditions. Larvae of C. hongkongensis were exposed to control pH8.0 and low pH7.4 conditions, mimicking near future OA scenario in their habitat, before being outplanted as post-metamorphic juveniles at two mariculture field sites with contrasting environmental stressors for 9months. The larval carry-over OA effect was found to have persistent impacts on the growth and survival trade-off traits on the outplanted juveniles, although the beneficial or adverse effect depended on the environmental conditions at the outplanted sites. Site-specific plasticity was demonstrated with a diverse DNA methylation-associated gene expression profile, with signal transduction and the endocrine system being the most common and highly enriched functions. Highly methylated exons prevailed in the key genes related to general metabolic and endocytic responses and these genes are evolutionarily conserved in various marine invertebrates in response to OA. These results suggest that oysters with prior larval exposure history to OA had the ability to trigger rapid local adaptive responses via epigenetic modification to cope with multiple stressors in the field.

Heavy Metal And Hydrocarbons Bioaccumulation By Two Bivalve’s Species From Santos Bay, Brazil

ABSTRACT Contaminants compounds like Heavy Metals and Polycyclic Aromatic Hydrocarbons (PAH’s) are primarily sourced from industrial and urban areas. As pollutants, these chemical elements and compounds are potentially toxic and present the capacity to negatively impact the ecosystem, especially the most sensitive ones such as estuaries. Bivalves’ species have been successfully utilized as monitoring tools to evaluate pollutants bioaccessibility around the globe. The present study assessed heavy metals (Zn, Cd, Ni, Fe, Cr, Cu and Pb), As and PAH’s levels in two mollusk species (Perna perna and Crassostrea rhizophorae) from Santos Estuarine System, in São Paulo State, in order to compare the effectiveness of each species in the biomonitoring process. According to the obtained data, Perna perna mussels may be better indicators of environmental pollution than Crassostrea rhizophorae oysters for some metals (Iron and Nickel) and hydrocarbons. On the other hand, in the present study, oysters had a higher capacity for retaining zinc, copper, and cadmium, due to their peculiar physiological characteristics.

Global controversy in oyster systematics and a newly described species from SE Asia (Bivalvia: Ostreidae: Crassostreinae)

The diversity of native oysters in many regions is overshadowed by the global dominance of a few economically important species. Here we describe the Muar Oyster, Crassostrea (Magallana) saidii sp. nov., first reported as an established local fishery renowned for exceptional and distinctive flavour over 160 years ago by British colonial officials in Malaysia, but as yet never formally named or described as a species. This new species has a subtle but clear morphological diagnosis dependent on three-dimensional characters, which has long been recognised by local fishers to differentiate the new species from co-occurring C. (M.) belcheri (G. B. Sowerby II, 1871). The Indo-Pacific clade Magallana Salvi & Mariottini, 2016 in Salvi and Mariottini 2017 is a phylogenetically distinct group that nonetheless cannot be morphologically separated from the broader genus Crassostrea Sacco, 1897. Fossils or species known only from shell specimens, though morphologically distinct species, cannot be classified as Magallana, Talonostrea Li & Qi, 1994, or Crassostrea s.s.; therefore, we revise these groups as sub-genera within Crassostrea. Our analysis of the COI barcoding fragment from previously published sequences of all available Magallana species found that gene is not sufficient to separate several economically important species, and revealed more than 5% of sequences in GenBank represent identification errors. The new species Crassostrea (Magallana) saidii sp. nov., which is genetically, morphologically, and gastronomically distinct, is known from only one population under potential threat from urbanisation. Many more global species of Magallana remain undescribed. The systematics of this group is critical to understand the diversity of global oysters, and to understand the sustainable use of species grown worldwide for centuries as our food.

Morphological and Mitochondrial DNA Analyses of Oysters in the Northern Bay of Bengal, Bangladesh

The geographic boundaries of many important habitat-building shallow estuarine oyster (Family Ostreidae) species are poorly understood, especially in subtropical and tropical waters. These keystone species often have extensive historical and extant ranges, in part because of their ability to adapt to diverse environmental conditions and the transfer and introduction of a few species worldwide for aquaculture production. In addition, oysters exhibit morphological plasticity additionally confounding species identification and taxonomy. Molecular techniques have led to significant improvements in oyster systematics and taxonomy but have not been applied to oysters from many tropical regions, including the coastal areas of the Indian Ocean such as Bangladesh. Because species' identification based on morphological traits alone, in this case primarily shell characteristics, were inadequate, phylogenetic analyses of mitochondrial cytochrome oxidase subunit I gene fragments was also done to confirm the identity of oyster specimens collected from Bangladesh coastal waters. Phylogenetic analyses of collected oyster samples confirmed the two monophyletic subclades of the Ostreidae family Crassostrea and Saccostrea, and five oyster lineages from the region of Bangladesh were identified: Crassostrea gryphoides, Crassostrea belcheri, Saccostrea cuccullata lineage B, S. cuccullata lineage F, and Saccostrea mordax. A new addition to the list of Crassostrea species, C. belcheri was found in Bangladesh, but C. belcheri, C. gryphoides, and S. cuccullata are considered “common” species in the neighboring states of India and Myanmar indicating a widespread distribution of these species along the entire coast of the Bay of Bengal. However, S. mordax, is a new record for the Bay of Bengal region including the coastal waters of Andaman and Nicobar Islands, and thus extends the geographical distribution of this species.

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.

Satellitome Analysis of the Pacific Oyster Crassostrea gigas Reveals New Pattern of Satellite DNA Organization, Highly Scattered across the Genome.

Several features already qualified the invasive bivalve species Crassostrea gigas as a valuable non-standard model organism in genome research. C. gigas is characterized by the low contribution of satellite DNAs (satDNAs) vs. mobile elements and has an extremely low amount of heterochromatin, predominantly built of DNA transposons. In this work, we have identified 52 satDNAs composing the satellitome of C. gigas and constituting about 6.33% of the genome. Satellitome analysis reveals unusual, highly scattered organization of relatively short satDNA arrays across the whole genome. However, peculiar chromosomal distribution and densities are specific for each satDNA. The inspection of the organizational forms of the 11 most abundant satDNAs shows association with constitutive parts of Helitron mobile elements. Nine of the inspected satDNAs are dominantly found in mobile element-associated form, two mostly appear standalone, and only one is present exclusively as Helitron-associated sequence. The Helitron-related satDNAs appear in more chromosomes than other satDNAs, indicating that these mobile elements could be leading satDNA propagation in C. gigas. No significant accumulation of satDNAs on certain chromosomal positions was detected in C. gigas, thus establishing a novel pattern of satDNA organization on the genome level.

Transgenerational responses to seawater pH in the edible oyster, with implications for the mariculture of the species under future ocean acidification

The majority of common edible oysters are projected to grow more slowly and have smaller impaired shells because of anthropogenic CO2-induced reductions in seawater carbonate ion concentration and pH, a process called ocean acidification (OA). Recent evidence has shown that OA has carryover effects, for example, larvae exposed to OA will also exhibit either positive or negative effects after metamorphosis. This study examined the hidden carryover effects of OA exposure during parental and larval stages on post-metamorphic traits of the commercially important oyster species Crassostrea hongkongensis. Adults of C. hongkongensis were exposed to control pH (pHNBS 8.0) and OA-induced low pH (pHNBS 7.4) conditions. Their larval offspring were then exposed to the same aquarium conditions before being out-planted as post-metamorphic juveniles at a mariculture site for 10 months. Initially, larval offspring were resilient to low pH with or without parental exposure. The larvae exposed to low pH had significantly faster development and higher percentage of settlement success compared to control groups. The out-planted juveniles with parental exposure had improved survival and growth compared to juveniles without parental exposure, regardless of the larval exposure history. This implies that transgenerational effects due to parental exposure not only persists but also have a greater influence than the within-generational effects of larval exposure. Our results shed light on the importance of linking the various life history stages when assessing the OA-induced carryover capacity of C. hongkongensis in the natural environment. Understanding these linked relationships helps us better predict the species rapid adaptation responses in the face of changing coastal conditions due to OA.

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.

The Sperm Proteome of the Oyster Crassostrea hongkongensis.

Sperm proteins play vital roles in fertilization, but little is known about their identities in free-spawning marine invertebrates. Here, 286 sperm proteins are reported from the Hong Kong oyster Crassostrea hongkongensis using label-free and semi-quantitative proteomics. Proteins extracted from three sperm samples are separated by SDS-PAGE, analyzed by LC-MS/MS, and identified using Mascot. Functional classification of the sperm proteome reveals energy metabolism (33%), signaling and binding (23%), and protein synthesis and degradation (12%) as the top functional categories. Comparison of orthologous sperm proteins between C. hongkongensis, Crassostrea gigas, Mytilus edulis, and M. galloprovincialis suggests that energy metabolism (48%) is the most conserved functional group. Sequence alignment of the C. hongkongensis bindin, an acrosomal protein that binds the sperm and the egg, with those of three other Crassostrea species, reveals several conserved motifs. The study has enriched the data of invertebrate sperm proteins and may contribute to studies of mechanisms of fertilization in free-spawning invertebrates. The proteomic data are available in ProteomeXchange with the identifier PXD018255.

Salinity and rainfall as inducers of cell proliferation and apoptosis in mangrove oyster Crassostrea gasar spermatogenesis

The present study investigated the influence of physical–chemical​ variables on the development of male gonads, providing information on cell proliferation and apoptosis in cells of the spermatogenic lineage throughout the reproductive cycle. Seeds of oysters of the species Crassostrea gasar from the same population were grown in two cultivation areas with different seawater influences. The area I, located in the municipality of Nova Olinda and area II, in the municipality of Curuçá. The two areas are part of the mangrove coast of the Amazon region and are located approximately 5 and 25 km away from the Atlantic Ocean, respectively. A sample containing 20 specimens of oysters was collected monthly in each of the two areas, and the abiotic parameters were also measured in situ during the collections. Routine histological techniques were used for cell characterization, as well as the classification of gonadal stages and morphometric measurements. Immunohistochemistry was used to access cell proliferation and apoptosis. The abiotic parameters varied constantly during the study, however, precipitation and salinity were the variables that most differed between periods in both areas. The cells of the spermatogenic lineage of C. gasar did not differ from the morphological point of view, however, the analysis of the morphometric parameters showed significant differences both in the cell diameters between maturation periods and between the study areas. Proliferative and apoptotic activities also varied throughout the study, and the quantification of positively labeled cells showed statistically significant differences between the study areas. The animals in the area I showed greater proliferative activity in relation to individuals in area II and using a regression model, we demonstrated that both cell proliferation and apoptosis are related to environmental conditions. It seems likely that animals in area I, exposed to a salinity of ∼25–35, find more favorable environmental conditions for their reproduction since the reduction in cell proliferation observed in area II was negatively related to the salinity of this environment (∼15– 25). Changes in the dynamics of mangrove interstitial waters contribute to changes in the salinity of the environment and can directly impact the physiology of oysters. These findings indicate that oyster farming systems in estuarine environments should be in areas as close to the ocean as possible.