Assessment of Gastroenteric Viruses in Marketed Bivalve Mollusks in the Tourist Cities of Rio de Janeiro, Brazil, 2022.

This study investigated the prevalence and genetic diversity of gastroenteric viruses in mussels and oysters in Rio de Janeiro, Brazil. One hundred and thirty-four marketed bivalve samples were obtained between January and December 2022. The viral analysis was performed according to ISO/TS 15216, and the screening revealed the detection of norovirus GII/GI (40.3%), sapovirus (SaV; 12.7%), human mastadenovirus (7.5%), and rotavirus A (RVA; 5.9%). In total, 44.8% (60) of shellfish samples tested positive for one or more viruses, 46.7% (28/60) of the positive samples tested positive for a single viral agent, 26.7% (16) tested positive for two viral agents, 8.3% (5) for three viral agents, and 13.3% (8) for four viral agents. Additionally, three mussel samples were contaminated with the five investigated viruses (5%, 3/60). Norovirus GII showed the highest mean viral load (3.4 × 105 GC/g), followed by SaV (1.4 × 104 GC/g), RVA (1.1 × 104 GC/g), human mastadenovirus (3.9 × 103 GC/g), and norovirus GI (6.7 × 102 GC/g). Molecular characterization revealed that the recovered norovirus strains belonged to genotypes GII.2, GII.6, GII.9, GII.17, and GII.27; SaV belonged to genotypes GI.1 and GIV.1; RVA to genotypes G6, G8, P[8]-III, and human mastadenovirus to types F40 and F41. The GII.27 norovirus characterized in this study is the only strain of this genotype reported in Brazil. This study highlights the dissemination and diversity of gastroenteric viruses present in commercialized bivalves in a touristic area, indicating the potential risk to human health and the contribution of bivalves in the propagation of emerging pathogens.

An international inter-laboratory study to compare digital PCR with ISO standardized qPCR assays for the detection of norovirus GI and GII in oyster tissue.

An optimized digital RT-PCR (RT-dPCR) assay for the detection of human norovirus GI and GII RNA was compared with ISO 15216-conform quantitative real-time RT-PCR (RT-qPCR) assays in an interlaboratory study (ILS) among eight laboratories. A duplex GI/GII RT-dPCR assay, based on the ISO 15216-oligonucleotides, was used on a Bio-Rad QX200 platform by six laboratories. Adapted assays for Qiagen Qiacuity or ThermoFisher QuantStudio 3D were used by one laboratory each. The ILS comprised quantification of norovirus RNA in the absence of matrix and in oyster tissue samples. On average, results of the RT-dPCR assays were very similar to those obtained by RT-qPCR assays. The coefficient of variation (CV%) of norovirus GI results was, however, much lower for RT-dPCR than for RT-qPCR in intra-laboratory replicates (eight runs) and between the eight laboratories. The CV% of norovirus GII results was in the same range for both detection formats. Had in-house prepared dsDNA standards been used, the CV% of norovirus GII could have been in favor of the RT-dPCR assay. The ratio between RT-dPCR and RT-qPCR results varied per laboratory, despite using the distributed RT-qPCR dsDNA standards. The study indicates that the RT-dPCR assay is likely to increase uniformity of quantitative results between laboratories.

Evaluation of a new automated viral RNA extraction platform for hepatitis A virus and human norovirus in testing of berries, lettuce, and oysters.

Fruits, vegetables, and shellfish are often associated with outbreaks of illness caused particularly by human norovirus (HuNoV) and hepatitis A virus (HAV), the leading causative agents of foodborne illness worldwide. The aim of this study was to evaluate a new automated nucleic acid extraction platform (EGENE-UP EASYPREP) for enteric viruses in several at-risk food matrices and to test its limit of detection in comparison to a semi-automated method (EGENE-UP) using Boom methodology for nucleic acid extraction as suggested in the reference method ISO 15216-2:2019. Fresh and frozen raspberries, frozen blackberries, romaine lettuce and oyster digestive glands were artificially contaminated with HAV, HuNoV GII.4 or HuNoV GI.7 at 102, 103 or 104 genome copies/sample. Virus was then recovered from the food matrix using the ISO method. Viral RNA extracted from frozen berry samples by the automated system was purified on a column for additional removal of RT-qPCR inhibitors. For fresh raspberry, oysters, and romaine lettuce, the two extraction platforms were deemed equivalent. For frozen raspberry, the automated platform appeared to be more efficient for viral recovery, particularly for HAV and HuNoV GI at lower concentrations. With frozen blackberries, the two platforms may be considered equivalent for all targeted viruses. However, the automated method led to less sample-associated inhibition of the PCR, 56.5 % of samples versus 95.0 % for the semi-automated. We thus found that the automated extraction can be performed easily by users while obtaining equivalent or even superior results to the ISO 15216-2:2019 method, and therefore appears to be suitable for routine sanitary monitoring in food processing and for tracing outbreaks of illness.

Pharmaceutical Residues in Edible Oysters along the Coasts of the East and South China Seas and Associated Health Risks to Humans and Wildlife.

The investigation of pharmaceuticals as emerging contaminants in marine biota has been insufficient. In this study, we examined the presence of 51 pharmaceuticals in edible oysters along the coasts of the East and South China Seas. Only nine pharmaceuticals were detected. The mean concentrations of all measured pharmaceuticals in oysters per site ranged from 0.804 to 15.1 ng g-1 of dry weight, with antihistamines being the most common. Brompheniramine and promethazine were identified in biota samples for the first time. Although no significant health risks to humans were identified through consumption of oysters, 100-1000 times higher health risks were observed for wildlife like water birds, seasnails, and starfishes. Specifically, sea snails that primarily feed on oysters were found to be at risk of exposure to ciprofloxacin, brompheniramine, and promethazine. These high risks could be attributed to the monotonous diet habits and relatively limited food sources of these organisms. Furthermore, taking chirality into consideration, chlorpheniramine in the oysters was enriched by the S-enantiomer, with a relative potency 1.1-1.3 times higher when chlorpheniramine was considered as a racemate. Overall, this study highlights the prevalence of antihistamines in seafood and underscores the importance of studying enantioselectivities of pharmaceuticals in health risk assessments.

Microplastics in rocky shore mollusks of different feeding habits: An assessment of sentinel performance.

Microplastics (MPs) accumulation in rocky shore organisms has limited knowledge. This study investigated MPs accumulation in filter-feeding oysters, herbivorous limpets and carnivorous snails to assess their performance as sentinel species in the MPs trophic transfer. The samples were obtained along a contamination gradient in the Santos Estuarine System, Brazil. All three studied species showed MPs concentrations related to the contamination gradient, being the oysters the species that showed the highest levels, followed by limpets and snails (average of less and most contaminated sites of 1.06-8.90, 2.28-5.69 and 0.44-2.10 MP g-1, respectively), suggesting that MPs ingestion rates are linked to feeding habits. MPs were mainly polystyrene and polyacetal. The polymer types did not vary among sites nor species. Despite minor differences in percentages and diversity of size, shape, and color classes, the analyzed species were equally able to demonstrate dominance of small, fiber, transparent, black and blue MPs. Thus, oysters, limpets, and snails are proposed as sentinels of MPs in monitoring assessments.

Seasonal profile of common pharmaceuticals in edible bivalve molluscs.

Pharmaceuticals are recognised as environmental contaminants of emerging concern (CECs) due to their increasing presence in the aquatic environment, along with high bioactivity linked to their therapeutic use. Therefore, information on environmental levels is urgently required. This study examined the presence of a range of common pharmaceuticals in oysters and mussels intended for human consumption from England and Wales using stable isotope dilution tandem mass spectrometry. A range of compounds were detected in bivalve tissue, with the Selective Serotonin Reuptake Inhibitor antidepressant sertraline being most abundant, reaching a maximum concentration of 22.1 ng/g wet weight shellfish tissue. Levels of all pharmaceuticals showed seasonal and geographical patterns. A dietary risk assessment revealed that the levels of pharmaceuticals identified in bivalve molluscs represent a clear hazard, but not a risk for the consumer. This study highlights the requirement for further monitoring of the presence of pharmaceuticals and other CECs in bivalve molluscs.

Complexity-functioning relationships differ across different environmental conditions.

Habitat complexity is widely considered an important determinant of biodiversity, and enhancing complexity can play a key role in restoring degraded habitats. However, the effects of habitat complexity on ecosystem functioning - as opposed to biodiversity and community structure - are relatively poorly understood for artificial habitats, which dominate many coastlines. With Greening of Grey Infrastructure (GGI) approaches, or eco-engineering, increasingly being applied around the globe, it is important to understand the effects that modifying habitat complexity has on both biodiversity and ecological functioning in these highly modified habitats. We assessed how manipulating physical (primary substrate) and/or biogenic habitat (bivalves) complexity on intertidal artificial substrata affected filtration rates, net and gross primary productivity (NPP and GPP, respectively) and community respiration (CR) - as well as abundance of filter feeders and macro-algae and habitat use by cryptobenthic fish across six locations in three continents. We manipulated both physical and biogenic complexity using 1) flat or ridged (2.5 cm or 5 cm) settlement tiles that were either 2) unseeded or seeded with oysters or mussels. Across all locations, increasing physical and biogenic complexity (5 cm seeded tiles) had a significant effect on most ecological functioning variables, increasing overall filtration rates and community respiration of the assemblages on tiles but decreasing productivity (both GPP and NPP) across all locations. There were no overall effects of increasing either type of habitat complexity on cryptobenthic fish MaxN, total time in frame or macro-algal cover. Within each location, there were marked differences in the effects of habitat complexity. In Hobart, we found higher filtration, filter feeder biomass and community respiration on 5 cm tiles compared to flat tiles. However, at this location, both macro-algae cover and GPP decreased with increasing physical complexity. Similarly in Dublin, filtration, filter feeder biomass and community respiration were higher on 5 cm tiles compared to less complex tiles. In Sydney, filtration and filter feeder biomass were higher on seeded than unseeded tiles, and fish MaxN was higher on 5 cm tiles compared to flat tiles. On unseeded tiles in Sydney, filter feeder biomass also increased with increasing physical complexity. Our findings suggest that GGI solutions via increased habitat complexity are likely to have trade-offs among potentially desired functions, such as productivity and filtration rates, and variable effects on cryptobenthic fish communities. Importantly, our results show that the effects of GGI practices can vary markedly according to the environmental context and therefore should not be blindly and uniformly applied across the globe.

Hazardous impacts of heavy metal pollution on biometric and biochemical composition of pearl oyster Pinctada radiata from five sites along Alexandria coast, with reference to its potential health risk assessment.

This study investigated the effect of heavy metals on the pearl oyster Pinctada radiata from 5 sites along the coast of Alexandria, with focus on its ecological health and potential risks to human consumption. Pollution results showed that Abu-Qir had the highest Cu and Cd values. Montaza and Eastern Harbor had the highest Fe and Pb values, respectively. Statistically, differences in metal concentrations among study sites were significant (p < 0.05). Non-carcinogenic risk (TTHQ) of tested metals and carcinogenic ones of Cd and Pb showed "high risk" on human health by consuming pearl oysters. Morphometric measurements and condition indices were studied to assess growth patterns and health in relation to heavy metals exposure. Key findings showed detectable declines in size and condition index in Eastern Harbor, whereas Abu-Qir recorded the highest values. This condition index performance presented Abu-Qir, Mammora, and Miami as ideal locations for spat collection and oyster rearing, potentially enhancing Egyptian pearl farming. Average values of spatial proximate contents of pearl oyster showed that it was rich in proteins (33.07-58.52%) with low fat content (1.39-1.87%) and carbohydrates (9.72-17.63%). Biochemical composition of pearl oyster demonstrated its high nutritional value which supported its promotion as a functional food for human consumption. The calorie content of pearl oyster was less than 2 Kcal, making this species an alternative source of healthy food to reduce obesity. Regression analysis indicated that Cu, Cd, and Pb had significant effect on 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity, calories, vitamins, and pigment content of the collected oysters.

Characterization of anthropogenic noise and oyster toadfish (Opsanus tau) calling behavior in urban and small-town coastal soundscapesa).

The oyster toadfish (Opsanus tau) is an ideal model to examine the effects of anthropogenic noise on behavior because they rely on acoustic signals for mate attraction and social interactions. We predict that oyster toadfish have acclimated to living in noise-rich environments because they are common in waterways of urban areas, like New York City (NYC). We used passive acoustic monitoring at two locations to see if calling behavior patterns are altered in areas of typically high boat traffic versus low boat traffic (Pier 40, NYC, NY, and Eel Pond, Woods Hole, MA, respectively). We hypothesized that toadfish in NYC would adjust their circadian calling behavior in response to daily anthropogenic noise patterns. We quantified toadfish calls and ship noise over three 24-h periods in the summer reproductive period at both locations. We observed an inverse relationship between the duration of noise and the number of toadfish calls at Pier 40 in comparison to Eel Pond. Additionally, toadfish at Pier 40 showed significant differences in peak calling behavior compared to Eel Pond. Therefore, oyster toadfish may have acclimated to living in an urban environment by potentially altering their communication behavior in the presence of boat noise.

A novel oyster shell biocomposite for the efficient adsorptive removal of cadmium and lead from aqueous solution: Synthesis, process optimization, modelling and mechanism studies.

This study highlights the effectiveness of oyster shell biocomposite for the biosorption of Cd(II) and Pb(II) ions from an aqueous solution. The aim of this work was to modify a novel biocomposite derived from oyster shell for the adsorption of Cd(II) and Pb(II) ions from aqueous solution. The studied revealed the specific surface BET surface area was 9.1476 m2/g. The elemental dispersive x-ray analysis (EDS) indicated that C, O, Ag, Ca were the predominant elements on the surface of the biocomposite after which metals ions of Cd and Pb were noticed after adsorption. The Fourier transform Irradiation (FT-IR) revealed the presence of carboxyl and hydroxyl groups on the surface. The effect of process variables on the adsorption capacity of the modified biocomposite was examined using the central composite design (CCD) of the response surface methodology (RSM). The process variables which include pH, adsorbent dose, the initial concentration and temperature were the most effective parameters influencing the uptake capacity. The optimal process conditions of these parameters were found to be pH, 5.57, adsorbent dose, 2.53 g/L, initial concentration, 46.76 mg/L and temperature 28.48°C for the biosorption of Cd(II) and Pb(II) ions from aqueous solution at a desirability coefficient of 1. The analysis of variance (ANOVA) revealed a high coefficient of determination (R2 > 0.91) and low probability coefficients for the responses (P < 0.05) which indicated the validity and aptness of the model for the biosorption of the metal ions. Experimental isotherm data fitted better to the Langmuir model and the kinetic data fitted better to the pseudo-second-order model. Maximun Cd(II) and Pb(II) adsorption capacities of the oyster shell biocomposite were 97.54 and 78.99 mg/g respectively and was obtained at pH 5.56 and 28.48°C. This investigation has provided the possibility of the utilization of alternative biocomposite as a sustainable approach for the biosorption of heavy metal ions from the wastewater stream.

Copper and zinc isotope systematics in different bivalve mollusk species from the French coastline: Implications for biomonitoring.

Zinc (Zn) and copper (Cu) stable isotopic compositions have been analyzed in various species of bivalve mollusks worldwide, but no comprehensive systematic interspecies comparison exists. Thus, we assessed isotope differences between species harvested in emblematic French coastal ecosystems to unveil biologically driven Cu and Zn isotope fractionation patterns. Inter-species isotopic variability of Cu is larger than Zn, with organisms that regulate internal concentrations displaying preferential bioaccumulation of heavy isotopes. The degree of internal isotope fractionation decreases from mussels > clams > oysters, affecting Cu more than Zn. The less pronounced Zn inter-specie variability helps preserve source information more reliably. Spatial analysis of a single oyster species denotes thus an important isotope variability of environmental Zn sources, including natural, anthropogenic and dietary components. Overall, results highlight the importance of considering systematic offset in Cu and Zn isotope values when comparing data from different bivalve species.

Effects of intensive oyster farming on nitrogen speciation in surface sediments from a typical subtropical mariculture bay.

The spatial-temporal distributions of various nitrogen (N) species in surface sediments were examined in a typical subtropical mariculture bay (Maowei Sea) in the northern Beibu Gulf to assess the impact of intensive oyster culture activities on sedimentary N speciation. The results indicated that the mean contents of total nitrogen (TN), extractable (labile) nitrogen (LN) and residual nitrogen (RN) in the surface sediments were 33.3 ± 15.5 µmol g-1, 13.8 ± 1.3 µmol g-1 and 19.5 ± 15.0 µmol g-1, respectively, which lacked significant seasonal variability (P > 0.05). Four forms of LN, namely ion extractable form (IEF-N), weak acid extractable form (WAEF-N), strong alkali extractable form (SAEF-N) and strong oxidant extractable form (SOEF-N) were identified based on sequential extraction. SOEF-N was the dominant form of LN, accounting for 67.8 ± 2.5 % and 63.7 ± 5.9 % in summer and winter, respectively. Spatially, the contents of sedimentary TN, LN, RN, WAEF-N and SOEF-N in intensive mariculture areas (IMA) were significantly higher than those in non-intensive mariculture areas (NIMA) during summer (P < 0.05). Stable nitrogen isotope (δ15N) mixing model revealed that shellfish biodeposition was the predominant source of sedimentary TN in IMA with a contribution of 67.8 ± 23.0 %, approximately 5.4 times that of NIMA (12.6 ± 3.3 %). Significant positive correlations (P < 0.05) were observed between most forms of N species (WAEF-N, SOEF-N, LN and RN) and shellfish-biodeposited N in the surface sediments during summer, indicating that intensive oyster farming greatly enhanced sedimentary TN accumulation.

Preparation of amino functionalized magnetic oyster shell powder adsorbent for selective removal of anionic dyes and Pb (II) from wastewater.

A kind of new magnetic oyster shell (OS) composite (OS-Fe3O4@SiO2@NH2@PEI) was synthesized and used as an adsorbent to remove carmine (CM), sunset yellow (SY) and Pb (II) from water. Firstly, Fe3O4 magnetic nanoparticles were introduced on the surface of waste oyster shell powder, then amino silanization was used to improve the stability of the material, and finally polyethylenimide (PEI) was grafted by Schiff base reaction. The composite was characterized by FT-IR, SEM, EDS, XPS, VSM, BET, TEM and zeta potential. The effects of adsorbent dosage and initial solution pH on the three samples were investigated by adsorption experiments. The adsorption kinetics and isotherms were investigated in depth under the best experimental conditions. The composite adsorbent not only selectively removed anionic azo dyes, but also had good recycling. In addition, OS-Fe3O4@SiO2@NH2@PEI still had good performance in mixed samples. These results indicated that OS-Fe3O4@SiO2@NH2@PEI was successfully used for the removal of a wide range of anionic dyes and heavy metal ions from the environment, and provided a new strategy for recycling waste.

Resilience against the impacts of climate change in an ecologically and economically significant native oyster.

Climate change is acidifying and warming our oceans, at an unprecedented rate posing a challenge for marine invertebrates vital across the globe for ecological services and food security. Here we show it is possible for resilience to climate change in an ecologically and economically significant oyster without detrimental effects to the energy budget. We exposed 24 pair-mated genetically distinct families of the Sydney rock oyster, Saccostrea glomerata to ocean acidification and warming for 4w and measured their resilience. Resilience was identified as the capacity to defend their acid-base balance without a loss of energy available for Scope for Growth (SFG). Of the 24 families, 13 were better able to defend their acid-base balance while eight had no loss of energy availability with a positive SFG. This study has found oyster families with reslience against climate change without a loss of SFG, is an essential mitigation strategy, in a critical mollusc.

Monitoring and Genotyping of Norovirus in Bivalve Molluscan Shellfish from Northern Italian Seas (2018-2020).

Norovirus (NoV) is an enteric virus with foodborne transmission. Bivalve shellfish are a main source of infections and outbreaks. In Italy a voluntary based monitoring plan to check the safety of bivalve shellfish was set up at provincial level. This study describes the occurrence and distribution of NoV in the Northern Adriatic Sea and in the Ligurian Sea. From October 2018 to September 2020, 807 bivalve shellfish samples (n = 205 oysters, n = 182 mussels, n = 348 clams, n = 72 other bivalve shellfish) were tested by One-Step Retrotranscription Real-time polymerase chain reaction for NoV GI and GII and quantified according to the ISO 15216-2:2013 and ISO 15216-1:2017. Positive samples were further analyzed to determine genotype by sequencing of the ORF1/ORF2 junction of the viral genome. A total of 126 samples were positive for NoV, mussels, and oysters had the highest probability of being positive and positive samples were found mainly in the colder season. Of these samples, 46% were NoV GII, 13% NoV GI, and 40% carried both genogroups. Thirty-seven samples were typeable (GI n = 12 and GII n = 25) with GI samples belonging to four genotypes and GII samples belonging to five genotypes. GII.3 genotype was the most prevalent, followed by GII.4, particularly Sydney 2012 subtype, a leading cause of infections worldwide, was found in three oysters' and three clams' samples. The phylogenetic analysis revealed a high heterogeneity among the species that are scattered in several clusters. Considering the low infectious dose the overall presence of NoV in edible shellfish, particular those to be eaten raw or undercooked, is moderately high. The presence of genotypes frequently involved in human infections strengthens the need for ongoing monitoring, which should be extended at national level.

Pearl Farming Micro-Nanoplastics Affect Oyster Physiology and Pearl Quality.

Pearl farming is crucial for the economy of French Polynesia. However, rearing structures contribute significantly to plastic waste, and the widespread contamination of pearl farming lagoons by microplastics has raised concerns about risks to the pearl industry. This study aimed to evaluate the effects of micro-nanoplastics (MNPs, 0.4-200 µm) on the pearl oyster (Pinctada margaritifera) over a 5-month pearl production cycle by closely mimicking ecological scenarios. MNPs were produced from weathered plastic pearl farming gear and tested at environmentally relevant concentrations (0.025 and 1 µg L-1) to decipher biological and functional responses through integrative approaches. The significant findings highlighted the impacts of MNPs on oyster physiology and pearl quality, even at remarkably low concentrations. Exposure to MNPs induced changes in energy metabolism, predominantly driven by reduced assimilation efficiency of microalgae, leading to an alteration in gene expression patterns. A distinct gene expression module exhibited a strong correlation with physiological parameters affected by MNP conditions, identifying key genes as potential environmental indicators of nutritional-MNP stress in cultured oysters. The alteration in pearl biomineralization, evidenced by thinner aragonite crystals and the presence of abnormal biomineral concretions, known as keshi pearls, raises concerns about the potential long-term impact on the Polynesian pearl industry.

Exploring PAH kinetics in wild vs. transplanted triploid and diploid oysters at a contaminated field site using immunological techniques.

Crassostrea virginica is a well-established bivalve species for biomonitoring persistent organic pollutants such as polycyclic aromatic hydrocarbons (PAH) in aquatic environments. Differing biomonitoring methods employing either wild oysters inhabiting sites of interest or naïve cultured oysters deployed to sites for extended periods can be used for site evaluations. However, important differences in total contaminant concentrations accumulated have been observed between the wild and transplanted groups. Furthermore, although rearing cultured triploid oysters is widely popular in commercial farming, the difference in contaminant bioaccumulation potential between triploid and diploid cultured oysters is vastly understudied, particularly for organic contaminants such as PAH. This study explores differences in PAH kinetics between transplanted triploid and diploid cultured oysters and wild oysters at a PAH-impacted site during a 6-week field exposure study using novel immunological techniques: antibody-based biosensor technology and immunofluorescence visualization. Conventional chemical analysis of oyster tissue was also conducted for comparison. While differences were observed in the oyster interstitial fluid between the wild and transplanted oysters throughout the study, whole tissue analysis revealed differing trends at each time point. Our findings suggest that insufficient equilibration time may contribute to the differences observed between groups. Furthermore, when combined with visual evidence via immunofluorescence, internal partitioning of contaminants may be an important determinant for total concentrations measured. A better understanding of the differences observed between wild and transplanted oyster groups is necessary for improved biomonitoring. Our study highlights the value in employing novel immunological techniques to explore possible mechanisms driving these differences.

Optimising flow-cytometry methods for marine mollusc haemocytes using the pearl oyster Pinctada maxima as a model.

Flow-cytometry has become increasingly popular to assess the haemocytes morphology and functions of marine molluscs. Indeed, haemocytes are the first line of defence of the immune system in molluscs and are used as a proxy for oyster health. Authors publishing in the field of flow-cytometry and molluscs health seemed to utilise the same methods for all model species used, independently of their geographical location in the world (temperate, tropical, etc.). Hence, this paper dived into flow-cytometry methodology and investigated if using different plates, different thresholds, different incubation times and temperatures as well as different fluorochromes concentrations affected the results. This study revealed that the cell count did not change when using different thresholds on the FSC-H parameter of the instrument but was affected by the plate type, the temperature of incubation, and the time of incubation. Indeed, non-adherent plates yielded the highest cell count and lower cell counts were associated with a higher temperature and a longer time of incubation. Furthermore, the haemocytes functions such as the phagocytosis, the lysosomal content, the intracellular oxidative activity, and the mitochondria activity were also affected by the temperature and the time of incubation. An increase in the phagocytosis capacity, lysosomal content and mitochondria activity was observed with a higher temperature. At the exception of the phagocytosis rate, all the other parameters such as the phagocytosis capacity, the intracellular oxidative activity, and the lysosomal content increased with a longer incubation time. We also showed that it is best to optimise the amount of fluorochromes used to avoid unnecessary background or non-specific staining.

Development of standards for assessing water quality in marine coastal waters of Bahrain.

Marine coastal waters of Bahrain are under pressure due to human activities and climate change. We used marine monitoring data (2005-2020) from 27 sites to establish baseline conditions and develop standards for assessments of water quality. Five hydrodynamic regions were identified: Oyster Beds, North, West, East, East (Coastal). Data from Oyster Beds sites, likely to be less impacted by human activities, were used to determine baseline conditions. For most parameters, candidate thresholds were based on 50 % and 100 % variation from baseline and 95th percentiles. Comparisons of data against different thresholds showed different outcomes. Overall, results indicate good water quality, with potential concerns in East (Coastal). Trend analyses showed some significant trends in all regions: downward (favourable) for some parameters (e.g. turbidity: North) and upward for others (e.g. nitrate: Oyster Beds, East and East (Coastal)). Future work requires greater understanding around optimum guidelines that protect and mitigate any adverse ecological impacts.

Oyster shell for drinking water filtration compared with granular activated carbon: advantages and limitations.

Deliberate media selection can be conducted to achieve targeted objective in filters. In this study, three biofilters (BFs) packed with calcinated oyster shell (COS), granular activated carbon (GAC), and COS + GAC (Mix) were set up in parallel following a rough filter packed with natural oyster shell to compare the performance for treating micro-polluted source water. Different media showed selective removal effects for different pollutants. GAC outperformed COS in terms of TOC and UV254. COS achieved higher reduction in turbidity than GAC. Due to the removal of total bacteria, the absolute and relative abundance of antibiotic resistance genes (ARGs) both decreased much in rough filter treated water (1.16 × 1014 to 1.40 × 1013 copies L-1 and 81.6 to 36.9%, respectively). The highest diverse and rich bacterial community was found in the biofilms on the COS filler, so microbial leakage gave rise to high bacterial content, leading to the highest absolute abundance of ARGs in COS BF effluent (2.11 × 1013 copies L-1). The highest relative abundance of ARGs (41.2%) was found in GAC BF effluent. SourceTracker and biomarker analysis both suggested that treatment process played a more important role in shaping the bacterial community structure in Mix BF effluent than single media BFs, which contributed to the lowest absolute (8.69 × 1012 copies L-1) and relative abundance (25.2%) of ARGs in Mix BF effluent among the three BFs. Our results suggested that mix COS + GAC can not only give full play to their respective advantages for traditional pollutants, but also achieve highest reduction in ARGs.