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Hemocytes are the first line of defense of the immune system in invertebrates, but despite their important role and enormous potential for the study of gene-environment relationships, research has been impeded by a lack of consensus on their classification. Here we used flow cytometry combined with histological procedures, histochemical reactions and transmission electron microscopy to characterize the hemocytes from the oyster Crassostrea rhizophorae. Transmission electron microscopy revealed remarkable morphological characteristics, such as the presence of membranous cisternae in all mature cells, regardless of size and granulation. Some granular cells contained many cytoplasmic granules that communicated with each other through a network of channels, a feature never previously described for hemocytes. The positive reactions for esterase and acid phosphatase also indicated the presence of mature cells of all sizes and granule contents. Flow cytometry revealed a clear separation in complexity between agranular and granular populations, which could not be differentiated by size, with cells ranging from 2.5 to 25 µm. Based on this evidence we suggest that, at least in C. rhizophorae, the different subpopulations of hemocytes may in reality be different stages of one type of cell, which accumulates granules and loses complexity (with no reduction in size) as it degranulates in the event of an environmental challenge.

Abstract The mangrove oyster Crassostrea rhizophorae is an estuarine resource exploited by riverside communities in Northeast Brazil. Despite its socioeconomic importance, studies on the health status of this bivalve are scanty in this region. The purpose of the present study was to investigate the presence of the protozoan Perkinsus sp. in C. rhizophorae collected in August and September 2011 in three estuaries of the septentrional Northeast, Brazil: Jaguaribe (Ceará), Camurupim (Piauí) and Carnaubeiras (Maranhão) (n= 150 specimens/site). The samples were submitted to Ray’s fluid thioglycollate medium (RFTM), PCR and histology assays. The RFTM assay revealed spherical, blue or bluish-black hypnospores of the genus Perkinsus in 50 specimens (Jaguaribe= 17.3%, Camurupim= 5.3%, Carnaubeiras= 10.6%). The intensity of the infection ranged from very light (1-10 cells per slide) to severe (more than 40 cells in each of 10 fields of the slide) for Jaguaribe; very light for Camurupim and very light to moderate (at least 40 cells observed in each of 10 fields of the slide) for Carnaubeiras. When submitted to confirmatory PCR analysis, 6 cases were confirmed (Jaguaribe=3, Camurupim=1, Carnaubeiras=2). The histology confirmed 21 cases of infection in specimens from the three estuaries. Although local collectors have reported no mortality in oyster populations that might be attributed to infection by Perkinsus, health surveillance of oyster populations in the septentrional region of Northeast Brazil is advisable. Resumo A ostra-do-mangue Crassostrea rhizophorae é um recurso estuarino explorado por comunidades ribeirinhas do Nordeste do Brasil. Apesar de sua importância socioeconômica, estudos sobre o estado de saúde deste bivalve são escassos na região. O objetivo deste estudo foi investigar a presença do protozoário Perkinsus sp. em C. rhizophorae coletada em agosto e setembro de 2011, em três estuários da região setentrional do Nordeste brasileiro: Jaguaribe (Ceará), Camurupim (Piauí) e Carnaubeiras (Maranhão) (n = 150 espécimes/local). As amostras foram submetidas ao meio líquido de tioglicolato de Ray (RFTM), PCR e ensaios histológicos. A análise em RFTM revelou hipnósporos esféricos azuis ou preto-azulados do gênero Perkinsus em 50 espécimes (Jaguaribe= 17,3%, Camurupim= 5,3%, Carnaubeiras= 10,6%). A intensidade de infecção variou de muito leve (1-10 células por lâmina) a severa (mais de 40 células em cada um dos 10 campos da lâmina) para o Rio Jaguaribe; muito leve para o Rio Camurupim e muito leve a moderada (pelo menos 40 células observadas, em cada um dos 10 campos da lâmina) para o Rio Carnaubeiras. Quando submetidos à análise confirmatória por PCR, foram confirmados 6 casos (Jaguaribe= 3, Camurupim= 1, Carnaubeiras= 2). A histologia confirmou 21 casos de infecção em espécimes dos três estuários. Embora os coletores locais não tenham relatado nenhuma mortalidade em populações de ostras que pudesse ser atribuída à infecção por Perkinsus, é aconselhável um monitoramento sobre o estado de saúde de populações de ostras da região.

Total mercury (Hg) was evaluated in the mangrove oyster Crassostrea rhizophorae, in sediments and seston from the Ciénaga Grande de Santa Marta (CGSM) and Cispatá Bay (BhC) in two climatic seasons (rainy and dry). Composite samples of sediments, seston and oysters in juvenile and adult ages were collected at six stations (three in each ecosystem) and Hg was quantified by atomic absorption spectrophotometry (Environmental Protection Agency (EPA) method 7473 PLTX-017). BhC had the highest Hg concentrations in sediment, seston and oysters compared to CGSM, with values close to the tolerable threshold for the ecosystem and associated biota (TEL) of 0.13 µg/g. Although at CGSM Hg was below the TEL in sediment and was considered safe in the oyster, significant bioconcentration was evident with the metal content in the seston, indicating a potential risk to the ecosystem and humans. The variables organic matter and temperature influenced metal availability in the sediment and seston, respectively; in contrast, they had no significant relationship in the oyster. In CGSM, higher Hg concentration was recorded in adult ages, while in BhC the highest accumulation occurred in juveniles, especially during the dry season. These findings underscore the importance of continuous Hg monitoring in both ecosystems. When assessed using the adapted Nemerow Pollution Index based on the provisional tolerable weekly intake (PTWI) of one µg Hg/kg body weight, although both sites presented a very high risk in terms of consumption, they are well below the most contaminated global hotspots over the past five decades. This study also highlights the relevance of oyster age in contamination assessments, as Hg accumulation patterns vary depending on environmental and climatic conditions.

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 of Crassostrea, as well as some congeneric species from other relevant areas. The Brazilian species are Crassostrea mangle sp.nov. and Crassostrea brasiliana. The exotic species Crassostrea 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, and Crassostrea 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 species Crassostrea gasar in Brazil is established as false. The voucher specimens of previous studies responsible for this assumption were examined, evidencing a misidentification for Crassostrea brasiliana from an estuarine environment. The geographical distribution of the studied species is recognized as follows: Crassostrea rhizophorae in the south-east Caribbean Sea; Crassostrea mangle sp. nov. from Pará to the Santa Catarina, only in mangroves; Crassostrea brasiliana from Paraíba to Santa Catarina, both in rocky shores and mangroves; Crassostrea virginica ranges from the Atlantic coast of Canada to the Caribbean; and Crassostrea gigas originates from the Indo-Pacific, but has been introduced in southern Brazil. This paper also deals with conchological aspects of the endemic species Crassostrea 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.

Objectives The two oyster species studied hold considerable economic importance for artisanal harvest ( Crassostrea rhizophorae ) and aquaculture ( Crassostrea gasar ). Their draft genomes will play an important role in the application of genomic methods such as RNAseq, population-based genomic scans aiming at addressing expression responses to pollution stress, adaptation to salinity and temperature variation, and will also permit investigating the genetic bases and enable marker-assisted selection of economically important traits like shell and mantle coloration and resistance to temperature and disease. Data description The draft assembly size of Crassostrea gasar is 506 Mbp, and of Crassostrea rhizophorae is 584 Mbp with scaffolds N50 of 11,3 Mbp and 4,9 Mbp, respectively. The general masked bases by RepeatMasker in both genomes were highly similar using different datasets. The masked bases varied from 9.41% in C. gasar to 10.05% in C. rhizophorae and 42.85% in C. gasar to 44.44% in C. rhizophorae using Dfam and RepeatModeler datasets, respectively. Functional annotation with eggNog resulted in 34,693 annotated proteins in C. rhizophorae and 26,328 in C. gasar . BUSCO analysis shows that almost 99% of genes (5,295) are complete in relation to the mollusk orthologous genes dataset (mollusca_odb10).

This paper aims to contribute to the use of mangrove cupped oyster, Crassostrea rhizophorae, as a biomonitor species for chemical contamination assessment in mangrove-lined Caribbean coastal systems. Sampling was carried out in eight localities (three in Nicaragua and five in Colombia) with different types and levels of contamination. Oysters were collected during the rainy and dry seasons of 2012–2013 and the tissue concentrations of metals, polycyclic aromatic hydrocarbons (PAHs), and persistent organic pollutants (POPs) were determined. Low tissue concentrations of metals (except Hg) and PAHs; moderate-to-high tissue concentrations of Hg, hexachlorocyclohexanes (HCHs), and dichlorodiphenyl-trichloroethanes (DDTs); detectable levels of chlorpyrifos, polychlorinated biphenyls (PCBs) (mainly CB28, CB118, CB138 and CB 153) and brominated diphenyl ethers 85 (BDE85); and negligible levels of musks were recorded in Nicaraguan oysters. A distinct profile of POPs was identified in Colombia, where the tissue concentrations of PCBs and synthetic musk fragrances were low to moderate, and Ag, As, Cd, Pb, and PAHs ranged from moderate to extremely high. Overall, the values recorded for HCHs, DDTs and PCBs in Nicaraguan mangrove cupped oysters greatly exceeded the reference values in tissues of C. rhizophorae from the Wider Caribbean Region, whereas only the levels of PCBs were occasionally surpassed in Colombia. Different contaminant profiles were distinguished between oysters from Nicaragua and Colombia in radar plots constructed using the main groups of contaminants (metals, PAHs, musks, PCBs, and organochlorine pesticides (OCPs)). Likewise, integrated pollution indices revealed differences in the levels of contaminants. Moreover, the profiles and levels in oyster tissues also varied with season. Thus, principal component analysis clearly discriminated Nicaraguan and Colombian localities and, especially in Colombia, seasonal trends in chemical contamination and differences amongst localities were evidenced. The geographical and environmental disparity of the studied scenarios may represent to a large extent the diversity of mangrove-lined Caribbean coastal systems and therefore the present results support the use of C. rhizophorae as suitable biomonitor species at Caribbean regional scale, where seasonal variability is a major factor controlling pollutant mobility and bioavailability.

The present study quantifies mercury (Hg) concentrations in mangrove oysters (Crassostrea rhizophorae) and assesses their potential as biomonitors of Hg contamination in the Parnaíba River Delta (PRD), located on the equatorial coast of Brazil (ECB). The highest Hg concentrations occurred in the smallest individuals’ size class (20–40 mm) from the main channel of the Parnaíba River (52.1 to 195.4 ng g−1 w.w.), whereas the largest individuals’ size class (larger than 60 mm) exhibited the lowest Hg concentrations (35.2–114 ng g−1 w.w.). There was a significant correlation between Hg concentrations and shell sizes, either when considering all size classes or when considering only individuals of size classes smaller than 40 mm. Oysters larger than 40 mm did not present any significant correlation between Hg concentrations and size. In addition to size, higher concentrations were observed at the freshwater–seawater transition in the main channel sites. These maximum suspended particulate zones, with bottom sediment resuspension, can favor Hg adsorption to fine particles, increasing the bioavailability of Hg. A regional comparison of Hg concentrations in mangrove oysters from the ECB suggests they are efficient biomonitors at a regional level. In contrast, the environmental dynamics of the PRD, with high variability within sites, hampered its use at the local level. Mangrove oysters from the PRD are shown to be safe for human consumption, as far as Hg exposure is concerned, and the presented risk assessment shows no excessive exposure, even at high-frequency consumption rates.

Mining in Colombia affects 488,672 ha (298,391 in coal mining and 190,281 in gold mining). However, Colombia has insufficient studies on mining and its repercussions, which limits estimates of mining impacts on ecosystems and the human population. Due to the rise of mining activities in Colombia, the negative impacts generated by coal and Hg will also continue to increase. This review analyzes national information levels on coal and Hg in island/coastal/marine as well as freshwater ecosystems and human groups using fishery resources as a framework. This is because fish are the main source of animal protein in marine coastal-island and mainland communities. Here, 15 of 32 Colombian departments have records on total mercury (THg) in water, sediments, fish, and human communities. Around 205 ton/year of mercury is discharged into the ecosystem. In human hair for example (15.3 to 50.15 μg/g), mercury exceeds the international maximum levels allowed (ILA) and the national standard (5.0 μg/g). Mercury levels in freshwater fish show 3.3 μg/g of THg and levels in marine and coastal-island fish are 1.2 μg/g THg exceeding the ILA (0.5 μg/g) standard for fish that will be consumed. Carnivorous species have a THg between 0.04 and 2.55 μg/g suggesting bioaccumulation and magnification of heavy metals. These findings were then compared with available international information.

Abstract This study reports the pathogen Perkinsus beihaiensis in oysters of the genus Crassostrea on the coast of the State of Bahia (Brazil), its prevalence, infection intensity and correlation with salinity. Oysters (n = 240) were collected between October and December 2014 at eight sampling stations between latitudes 13°55'S and 15°42'S. The laboratory procedures included macroscopic analysis, histology, culture in Ray's fluid thioglycollate medium (RFTM), Polymerase Chain Reaction (PCR) and DNA sequencing. PCR and sequencing have been used for the genetic identification of oysters as well. Two species of oysters have been identified: Crassostrea rhizophorae and C. brasiliana. In both oyster species P. beihaiensis was the only Perkinsus species detected. In C. rhizophorae, the average prevalence was 82.8% by histology and 65.2% by RFTM. In C. brasiliana, the prevalences were 70.5% and 35.7%, respectively. The higher prevalence of P. beihaiensis in C. rhizophorae was probably influenced by salinity, with which was positively correlated (r> 0.8). In both oysters, P. beihaiensis was located mainly in the gastric epithelium. The infection was generally mild or moderate, without apparent harm to the hosts, but in cases of severe infection, there was hemocytical reaction and tissue disorganization. The generally high prevalence in the region suggests that oysters should be monitored with respect to this pathogen, especially in growing areas. Resumo Este estudo relata o patógeno Perkinsus beihaiensis em ostras do gênero Crassostrea no litoral do Estado da Bahia (Brasil), sua prevalência, intensidade de infecção e correlação com a salinidade. As ostras (n = 240) foram coletadas entre outubro e dezembro de 2014 em oito estações amostrais entre as latitudes 13°55'S e 15°42'S. Os procedimentos laboratoriais incluíram análise macroscópica, histologia, cultivo em meio de tioglicolato de Ray (RFTM), reação em cadeia da polimerase (PCR) e sequenciamento de DNA. PCR e sequenciamento foram também utilizados para a identificação genética das ostras. Foram identificadas duas espécies de ostras: Crassostrea rhizophorae e C. brasiliana. Em ambas as espécies, P. beihaiensis foi a única espécie de Perkinsus detectada. Em C. rhizophorae, a prevalência média foi de 82,8% por histologia e de 65,2% por RFTM. Em C. brasiliana, as prevalências foram de 70,5% e 35,7%, respectivamente. A maior prevalência de P. beihaiensis em C. rhizophorae foi provavelmente influenciada pela salinidade, com a qual este apresentou correlação positiva (r>0,8). Em ambas as espécies, P. beihaiensis esteve localizada principalmente no epitélio gástrico. A infecção foi geralmente leve ou moderada, sem danos aparentes aos hospedeiros, mas em casos de infecção severa, houve reação hemocitária e desorganização de tecidos. As prevalências geralmente altas na região sugerem que as ostras devam ser monitoradas com relação a este patógeno, principalmente em áreas de cultivo.