Characterisation of the Bacteria and Archaea Community Associated with Wild Oysters, At Three Possible Restoration Sites in the North Sea

With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including microorganisms that drive marine biogeochemical cycles. This experiment established a baseline for the monitoring of the bacterial and archaeal community associated with wild oysters, using samples from their immediate environment of the Voordelta, with cohabiting Crassostrea gigas and Ostrea edulis, Duikplaats with only C. gigas attached to rocks, and the Dansk Skaldyrcentre, with no onsite oysters. The microbial profiling was carried out through DNA analysis of samples collected from the surfaces of oyster shells and their substrate, the sediment and seawater. Following 16S rRNA amplicon sequencing and bioinformatics, alpha indices implied high species abundance and diversity in sediment but low abundance in seawater. As expected, Proteobacteria, Bacteroidetes, Firmicutes and Thaumarchaeota dominated the top 20 OTUs. In the Voordelta, OTUs related to Colwellia, Shewanella and Psychrobium differentiated the oysters collected from a reef with those attached to rocks. Duikplaats were distinct for sulfur-oxidizers Sulfurimonas and sulfate-reducers from the Sva 0081 sediment group. Archaea were found mainly in sediments and the oyster associated microbiome, with greater abundance at the reef site, consisting mostly of Thaumarchaeota from the family Nitrosopumilaceae. The oyster free site displayed archaea in sediments only, and algal bloom indicator microorganisms from the Rhodobacteraceae, Flavobacteriaceae family and genus [Polaribacter] huanghezhanensis, in addition to the ascidian symbiotic partner, Synechococcus. This study suggests site specific microbiome shifts, influenced by the presence of oysters and the type of substrate.

Detection and quantification of Pb and Cr in oysters using laser-induced breakdown spectroscopy

The quantitative determination of heavy metals in aquatic products is of great importance for food security issues.Laser-induced breakdown spectroscopy(LIBS)has been used in a variety of foodstuff analysis,but is still limited by its low sensitivity when targeting trace heavy metals.In this work,we compare three sample enrichment methods,namely drying,carbonization,and ashing,for increasing detection sensitivity by LIBS analysis for Pb and Cr in oyster samples.The results demonstrate that carbonization can remove a significant amount of the contributions of organic elements C,H,N and O;meanwhile,the signals of the metallic elements such as Cu,Pb,Sr,Ca,Cr and Mg are enhanced by3–6 times after carbonization,and further enhanced by 5–9 times after ashing.Such enhancement is not only due to the more concentrated metallic elements in the sample compared to the dried ones,but also the unifying of the matter in carbonized and ashed samples from which higher plasma temperature and electron density are observed.This condition favors the detection of trace elements.According to the calibration curves with univariate and multivariate analysis,the ashing method is considered to be the best choice.The limits of detection of the ashing method are 0.52 mg kg-1 for Pb and0.08 mg kg-1 for Cr,which can detect the presence of heavy metals in the oysters exceeding the maximum limits of Pb and Cr required by the Chinese national standard.This method provides a promising application for the heavy metal contamination monitoring in the aquatic product industry.

论Oysters

林奈于1758年建立了Ostrea (s.l.)属。此后的两百多年中经拉马克(Lamark)、杜威尔(Douville)、乌雅洛夫(Vyalov)、斯坦兹尔(Stenzel)等许多学者的深入研究,不断修正、补充与完善,原来的Oatrea (s.l.)属现已扩展为牡蛎超科(SuperfamilyOstreacea),人们通常称之为Oysters(牡蛎类)。目前对Oysters的特征、分类、生态、起源、演化等方面的认识正不断深化,但仍存在不少分歧。

High Pressure Treatment Changes Spoilage Characteristics and Shelf Life of Pacific Oysters(Crassostrea gigas)During Refrigerated Storage

The effects of high pressure(HP) treatment on spoilage characteristic and shelf life extension of Pacific oysters(Crassostrea gigas) during refrigerated storage were studied.Results showed that HP treatment of 275 MPa for 3 min or 300 MPa for 2 min could achieve 100% full release of oyster adductor muscle,pressures higher than 350 MPa caused excessive release as the shells of oysters were broken,thus use of higher pressures should be cautious in oyster processing industry because of its adverse impact on the appearance of shells.HP treatment(300 MPa,2 min) was proper for the shucking of Pacific oyster(Crassostrea gigas) in China.This treatment caused no organoleptic disadvantage.Moreover,HP treatment resulted in obvious differences in biochemical spoilage indicators(p H,TVB-N and TBARS) changes and volatile compounds profile determined by electronic nose during storage.HP treatment(300 MPa,2 min) also led to a reduction of aerobic bacterial count(APC) by 1.27 log cycles.Furthermore,the APC values of oysters treated by HP were always lower than those of the control samples during storage.Based on the organoleptic,biochemical and microbiological indicators,shelf life of 6-8 d for control and 12 d for HP-treated oysters could be expected.HP treatment showed great potential in oyster processing and preservation.

Occurrence of Toxigenic Clostridium difficile in Louisiana Oysters (<i>Crassostrea virginica</i>) and Environmental Waters

Clostridium difficile infection (CDI) was generally considered as a hospital-associated disease;however, recent community-acquired CDI has raised the concerns regarding the transmission of the pathogen through environmental sources. Limited data are available regarding the presence of C. difficile in food and water. In this study, oysters (Crassostrea virginica) and the harvest water collected from the commercial harvesting areas along the Louisiana Gulf Coast as well as the influent and effluent of a municipal treatment plant in New Orleans, LA were analyzed for toxigenic C. difficile. The bacterium was isolated from 47.37% (9/19) of oysters and 37.5% (3/8) of harvest water samples. Toxigenic C. difficile were also detected in all the wastewater influent and effluent samples. All the isolates harbored the gene tcdB encoding the virulence factor toxin B. Further PCR-ribotyping showed that the C. difficile isolated from the oysters and harvest water differed from the wastewater isolates. However, similar ribotypes were found in oysters and the surrounding harvest water. We found that oysters growing in contaminated water could bioaccumulate toxigenic C. difficile and pose a health risks by serving as a vehicle for the transmission of the pathogen to humans.

Selection of Reference Genes for Expression Analysis of Kumamoto and Portuguese Oysters and Their Hybrid

Quantitative real-time polymerase chain reaction(q RT-PCR) is a rapid and reliable technique which has been widely used to quantifying gene transcripts(expression analysis). It is also employed for studying heterosis, hybridization breeding and hybrid tolerability of oysters, an ecologically and economically important taxonomic group. For these studies, selection of a suitable set of housekeeping genes as references is crucial for correct interpretation of q RT-PCR data. To identify suitable reference genes for oysters during low temperature and low salinity stresses, we analyzed twelve genes from the gill tissue of Crassostrea sikamea(SS), Crassostrea angulata(AA) and their hybrid(SA), which included three ribosomal genes, 28 S ribosomal protein S5(RPS5), ribosomal protein L35(RPL35), and 60 S ribosomal protein L29(RPL29); three structural genes, tubulin gamma(TUBγ), annexin A6 and A7(AA6 and AA7); three metabolic pathway genes, ornithine decarboxylase(OD), glyceraldehyde-3-phosphate dehydrogenase(GAPDH) and glutathione S-transferase P1(GSP); two transcription factors, elongation factor 1 alpha and beta(EF1α and EF1β); and one protein synthesis gene(ubiquitin(UBQ). Primers specific for these genes were successfully developed for the three groups of oysters. Three different algorithms, ge Norm, Norm Finder and Best Keeper, were used to evaluate the expression stability of these candidate genes. Best Keeper program was found to be the most reliable. Based on our analysis, we found that the expression of RPL35 and EF1α was stable under low salinity stress, and the expression of OD, GAPDH and EF1α was stable under low temperature stress in hybrid(SA) oyster; the expression of RPS5 and GAPDH was stable under low salinity stress, and the expression of RPS5, UBQ, GAPDH was stable under low temperature stress in SS oyster; the expression of RPS5, GAPDH, EF1β and AA7 was stable under low salinity stress, and the expression of RPL35, EF1α, GAPDH and EF1β was stable under low temperature stress in AA oyster. Furthermore, to evaluate their suitability, the reference genes were used to quantify six target genes. In conclusion, we have successfully developed primers appropriate for the expression analysis in SS, SA and AA.

Acid Sulfate Soil Induced Acidification of Estuarine Areas Used for the Production of Sydney Rock Oysters, <i>Saccostrea glomerata</i>

This study investigated estuarine acidification, associated with drainage and excavation of acid sulfate soils, in areas used for commercial cultivation of Sydneyrock oysters (Saccostrea glomerata). Regular measurements of pH and electrical conductivity were collected in oyster cultivation areas and acidified reaches of the Hastings River estuary and Port Stephens estuary located on the mid north coast of New South Wales, Australia. Water quality information from acidified floodplain drains was also collected in theHastingsRiverfollowing heavy rainfall. Both estuaries experienced acidification of tributaries following periods of heavy rainfall. Drain outflow waters were acidic

Factors Affecting Dermo Disease(Perkinsus marinus)in Eastern Oysters(Crassostrea virginica)in Galveston Bay,Texas

The oyster disease Dermo(Perkinsus marinus)affects the viability of oyster reefs of Galveston Bay,Texas.Documenting the relationships between distribution and prevalence of Dermo disease in the eastern oyster(Crassostrea virginica)and environmental conditions is beneficial to management of the eastern oyster in Galveston Bay.We sampled four sites located in Galveston Bay every other month from November 2014 through September 2015.The focus of the study was the relationship of water quality parameters(i.e.,fresh-water flow,salinity,water temperature,and water turbidity)to prevalence and parasite concentration of Dermo disease in oysters.Dermo was present in oysters at all reefs sampled,and Dermo prevalence was greatest at April Fool and Confederate reefs,but declined after heavy rainfall.Linear regression analysis indicated water variables such as temperature,salinity,turbidity,and fresh water inflow explained different amounts of the variability in the Mackin Dermo Intensity Scale among sampled reefs.We found combinations of low fresh-water inflow,high salinity,and high temperatures accounted for majority of the variance of Dermo in oysters located in Galveston Bay.

Three novel umami peptides derived from the alcohol extract of the Pacific oyster(Crassostrea gigas):identification,characterizations and interactions with T1R1/T1R3 taste receptors

Oyster(Crassostrea gigas),the main ingredient of oyster sauce,has a strong umami taste.In this study,three potential umami peptides,FLNQDEEAR(FR-9),FNKEE(FE-5),and EEFLK(EK-5),were identified and screened from the alcoholic extracts of the oyster using nano-HPLC-MS/MS analysis,i Umami-Scoring Card Method(i Umami-SCM)database and molecular docking(MD).Sensory evaluation and electronic tongue analysis were further used to confirm their tastes.The threshold of the three peptides ranged from 0.38 to 0.55 mg/m L.MD with umami receptors T1R1/T1R3 indicated that the electrostatic interaction and hydrogen bond interaction were the main forces involved.Besides,the Phe592 and Gln853 of T1R3 were the primary docking site for MD and played an important role in umami intensity.Peptides with two Glu residues at the terminus had stronger umami,especially at the C-terminus.These results contribute to the understanding of umami peptides in oysters and the interaction mechanism between umami peptides and umami receptors.

Non-Targeted Metabolomics Reveals the Metabolic Alterations in Response to Artificial Selective Breeding in the Fast-Growing Strains of Pacific Oyster

Pacific oyster(Crassostrea gigas)is one of the most important mollusks cultured all around the world.Selective breeding programs of Pacific oysters in China is initiated since 2006 and developed the genetically improved strain with fast-growing trait.However,little is known about the metabolic signatures of the fast-growing trait.In the present study,the non-targeted metabolomics was performed to analyze the metabolic signatures of adductor muscle tissue in one-year old Pacific oysters from fast-growing strain and the wild population.A total of 7767 and 10174 valid peaks were extracted and quantified in ESI^(+)and ESI^(−)modes,resulting in 399 and 381 annotated metabolites,respectively.PCA and OPLS-DA revealed that considerable separation among samples from fastgrowing strain and wild population,suggesting the differences in metabolic signatures.Meanwhile,81 significantly different metabolites(SDMs)were identified in the comparisons between fast-growing strain and wild population,based on the strict thresholds.It was found that there were highly correlation and conserved coordination among these SDMs.KEGG enrichment analysis indicated that the SDMs were tightly related to pantothenate and CoA biosynthesis,steroid hormone biosynthesis,riboflavin metabolism,and arginine and proline metabolism.Of them,the CoA biosynthesis and metabolism,affected by pantetheine and pantothenic acid,might be important for the growth of Pacific oysters under artificial selective breeding.The study provides the comprehensive views of metabolic signatures in response to artificially selective breeding,and is helpful to better understand the molecular mechanism of fastgrowing traits in Pacific oysters.

Use of natural trophic resources by Eastern oysters and Pacific oysters of different ploidy

Ploidy manipulation,such as triploidy,in farmed oysters has been used as a tool to enhance oyster quality throughout all seasons because triploid oysters allocate less energy to gametogenesis,and therefore are deemed better performers than fertile diploids.Nevertheless,scientific reports describing no differences between ploidies and,in certain conditions,disadvantages of triploids are not uncommon.As the use of triploid oyster seed increases culture cost for growers,consistency in performance of triploids is considered to be an important goal.Thus,research to assess how ploidy affects physiological processes underlying oyster performance is fundamental for the aquaculture industry.This work was undertaken to assess if ploidy-based differences in performance in the two most commonly cultivated commercial species of oyster in the United States,Crassostrea virginica and Crassostrea gigas,are associated with filtration,feeding,and metabolism.To test this hypothesis,biodeposition measurements were made with oysters exposed to ambient water conditions at locations and seasons providing a variety of environmental conditions.Oysters did not show differences in filtration and feeding associated with ploidy,but physiological feeding variables fluctuated with environmental characteristics associated with spatial and seasonal differences.A preliminarily test of the hypothesis that differences in energy metabolism may account for differences in performance among ploidy levels indicates that basal metabolic rates of diploid,triploid,and tetraploid Eastern oysters are not different.

Photoinactivation of bacteriophage MS2,Tulane virus and Vibrio parahaemolyticus in oysters by microencapsulated rose bengal

Bivalve molluscan shellfish such as oysters are important vectors for the transmission of foodborne pathogens including both viruses and bacteria.Photoinactivation provides a cold-sterilization option against the contamination as excited photosensitizers could transfer electronic energy to oxygen molecules producing reactive oxygen species such as singlet oxygen,leading to oxidative damage and death of the pathogens.However,the efficacy of photoinactivation is very often compromised by the presence of food matrix due to the nonselective reactions of short-lived singlet oxygen with organic matter other than the target pathogens.In order to address this issue,we encapsulated a food-grade photosensitizer rose bengal(RB)in alginate microbeads.An extra coating of chitosan effectively prevented the release of RB from the microbeads in seawater,and more importantly,enhanced the selectivity of the photoinactivation via the electrostatic interactions between cationic chitosan and anionic charge of the virus particles(bacteriophage MS2 and Tulane virus)and the Gram-negative bacteria Vibrio parahaemolyticus(V parahaemolyticus).The treatment of oysters with microencapsulated RB resulted in significantly higher reductions of MS2 phage,Tulane virus and V parahaemolyticus than free RB and non-RB carrying microbeads(P<0.05)tested with both in vitro and in vivo experimental set-ups.This study demonstrated a new strategy in delivering comprehensively formulated biochemical sanitizers in bivalve shellfish through their natural filter-feeding activity and thereby enhancing the mitigation efficiency of foodborne pathogen contamination.

Contraction of Heat Shock Protein 70 Genes Uncovers Heat Adaptability of Ostrea denselamellosa

The milin oyster Ostrea denselamellosa is a live-bearing species with a sharp decline in the natural population.Unlike other oysters,O.denselamellosa lives in the subtidal zone and its adaptability to heat,salinity,etc.is different from most other oys-ters.Heat shock proteins 70(HSP70)are a family of conserved ubiquitously expressed heat shock proteins which are produced in re-sponse to stressful conditions,especially heat.In this study,we identified Hsp70 genes through bioinformatic analysis in five species of oyster.Among them,O.denselamellosa holds the fewest number of Hsp70 genes,which may be one of the reasons why O.den-selamellosa cannot tolerate high temperatures.The conserved motifs and gene structures of Hsp70B2 sub-family and other types of Hsp70 in O.denselamellosa were different from that of Hspa12 sub-family,which may be due to performing necessary multiple phy-siological functions.Transcription profile analysis for Hsp70 genes of O.denselamellosa indicated that gills play an important role in responding to multiple external challenges.In addition,synteny analysis of Hsp70 genes among O.denselamellosa,O.edulis and Cras-sostrea ariakensis showed that Hsp70 genes in genus of Ostrea genome might have evolved from a common ancestor with genus of Crassostrea.In short,our results lay the foundation for further investigation of the evolution of O.denselamellosa Hsp70 genes and heat adaptability.

Biological oyster shell waste enhances polyphenylene sulfide composites and endows them with antibacterial properties

To date,there is no research that deals with biological waste as fillers in polyphenylene sulfide(PPS).In this study,oyster shells were recycled and treated to prepare thermally-treated oyster shells(TOS),which were used as PPS fillers to make new bio-based antibacterial composite materials.The effect of varying the content of TOS was studied by means of structure and performance characterization.PPS/TOS composites were demonstrated to have an antibacterial effect on the growth of E coli and S.aureus.Qualitative analysis showed that when the TOS content was≥30%and 40%,the composite materials had an apparent inhibition zone.Quantitative analysis showed that the antibacterial activity increased with the TOS content.Fourier transform infrared spectroscopy indicated the formation of hydrogen bonds between the molecular chains of TOS and PPS and the occurrence of a coordination reaction.At 10%TOS,the composite tensile strength reached a maximum value of 72.5 MPa,which is 9.65%higher than that of pure PPS.The trend of bending properties is the same as that of tensile properties,showing that the maximum property was reached for the composite with 10%TOS.At the same time,the crystallinity and contact angle were the highest,and the permeability coefficient was the lowest.The fatigue test results indicated that for the composite with 10%TOS,the tensile strength was 23%lower than static tensile strength,and the yield strength was 10%lower than the static yield strength.The results of the study showed that TOS not only could reduce the cost of PPS,but also could impart antibacterial properties and enhance the mechanical and,barrier properties,the thermostability,as well as the crystallinity.

Effects of shellfish culture on the community and mortality of zooplankton in a subtropical Bay

Spatial and temporal distribution of abundance,community structure and succession,and mortality of zooplankton were examined in a subtropical bay—the Qinzhou Bay,Guangxi,South China,to evaluate the impact of oyster culture on zooplankton.Data on zooplankton abundance,phytoplankton biomass,and environmental parameters were collected in 12 routine samplings at four sites in the Qinzhou Bay in 2018.Results show that the hydrological environment in the estuary affected the temporal variation of zooplankton of the Qinzhou Bay.However,the oyster culture affected the spatial distributions of zooplankton.The abundance of zooplankton outside the oyster farm was higher than that inside,whereas the mortality outside the oyster farm,on the contrary,was lower than that inside.Oyster culture in the Qinzhou Bay had built a significant impact on the abundance and community composition of zooplankton,and the food shortage caused by the ingestion of oyster might exacerbate the mortality of zooplankton in the Qinzhou Bay.In addition,the large-scale oyster culture increased the spatial heterogeneity of the community structure of zooplankton inside and outside the oyster farms.Findings of this study provide a reference for future policy-making in aquaculture management.

Near-infrared spectroscopy method for rapid proximate quantitative analysis of nutrient composition in Pacific oyster Crassostrea gigas

Glycogen,amino acids,fatty acids,and other nutrient components affect the flavor and nutritional quality of oysters.Methods based on near-infrared reflectance spectroscopy(NIRS)were developed to rapidly and proximately determine the nutrient content of the Pacific oyster Crassostreagigas.Samples of C.gigas from 19 costal sites were freeze-dried,ground,and scanned for spectral data collection using a Fourier transform NIR spectrometer(Thermo Fisher Scientific).NIRS models of glycogen and other nutrients were established using partial least squares,multiplication scattering correction first-order derivation,and Norris smoothing.The R_(C) values of the glycogen,fatty acids,amino acids,and taurine NIRS models were 0.9678,0.9312,0.9132,and 0.8928,respectively,and the residual prediction deviation(RPD)values of these components were 3.15,2.16,3.11,and 1.59,respectively,indicating a high correlation between the predicted and observed values,and that the models can be used in practice.The models were used to evaluate the nutrient compositions of 1278 oyster samples.Glycogen content was found to be positively correlated with fatty acids and negatively correlated with amino acids.The glycogen,amino acid,and taurine levels of C.gigas cultured in the subtidal and intertidal zones were also significantly different.This study suggests that C.gigas NIRS models can be a cost-effective alternative to traditional methods for the rapid and proximate analysis of various slaughter traits and may also contribute to future genetic and breeding-related studies in Pacific oysters.

The dual-function of bioactive peptides derived from oyster (Crassostrea gigas) proteins hydrolysates

Oysters(Crassostrea gigas)have a wide range of functionality due to their nutritional and bioactive components. However, the bioactive peptides of oyster proteins are rarely reported, particularly their antidiabetes effects and antioxidants. Oyster proteins were extracted from fresh oysters using phosphatebuffered saline and simulated gastrointestinal digestion was performed. The degree of hydrolysis(DH), structural characterization, molecular weight(Mw)distribution, free amino acid, anti-diabetic activity, and antioxidant activity were studied during in vitro simulated gastrointestinal digestion. The results showed that the α-glucosidase inhibitory activity, α-amylase inhibitory activity, DPPH radical scavenging activity, and ABTS radical scavenging activity of the oyster protein gastrointestinal digest were increased(P < 0.05)from 0 to 33.96%, from 9.17% to 44.22%, from 9.01 μg trolox/mg protein to 18.48 μg trolox/mg protein, and from 21.44 μg trolox/mg protein to 56.21 μg trolox/mg protein, respectively. Additionally, the DH, β-turn structure, fluorescence intensity, free amino acid, and short peptide content(Mw < 1 000 Da)increased in the simulated gastrointestinal digestion. These results indicate that the digestive hydrolysates obtained from oyster proteins could be used as natural anti-diabetic and antioxidant agents.

Preparation of Iron-Pillared Bentonite/Oyster Shell Composite and Phosphate Adsorption in Water

Iron-pillared bentonite(FB)was prepared by Fe(III)modified bentonite,and then the composites(FB-OS)were prepared by iron-pillared bentonite and oyster shell powder.The composites were characterized by FTIR,SEM,TGA,and EDS,and the phosphorus removal test was carried out.The results showed that FB-OS contained a large amount of CaO.Its structure was compact,but there were gaps in it.The maximum bending stress and compressive strength were 43.7 N and 0.927 MPa,respectively.The phosphorus removal test showed that the phosphorus removal rate of FB-OS was more than 90%,and measured the maximum adsorption capacity was 48.31 mg/g.A large amount of spherical products were produced on the surface and inside of FB-OS after phos-phorus removal,it was speculated that spherical products were amorphous calcium phosphate in the paper.Ana-lysis indicated that there was chemical adsorption during phosphorus removal.The kinetic equation of phosphorus adsorption by FB-OS was qt=10:193t/1+2:574t (R^(2)=0.995).The adsorption rate was mainly controlled by outerfilm diffusion and intraparticle diffusion.

Purification of β-Glucan of Oyster Mushroom (Pleurotus pulmonarius) and Its Application in Model Food

Mushrooms are recognized as sources of β-glucan. The aim of this study was to purify β-glucan from oyster mushrooms (Pleurotus pulmonarius) by alkaline extraction. The extracted β-glucan was added to a cracker recipe and the sensory, physical and estimated glycemic index of the final product were evaluated. Optimum extraction parameters were determined to be 80˚C, 90 minutes and 30% KOH. The β-glucan crackers had a glycemic index ranging from 88.07 to 67.32, while glucose had a glycemic index of 100. Based on the sensory study, it can be concluded that crackers enriched with β-glucan and possessing functional qualities may meet consumer expectations.

Electrical Stimulation Greatly Increases Settlement, Growth, Survival, and Stress Resistance of Marine Organisms

Increasing stress from global warming, sea level rise, acidification, sedimentation, pollution, and unsustainable practices have degraded the most critical coastal ecosystems including coral reefs, oyster reefs, and salt marshes. Conventional restoration methods work only under perfect conditions, but fail nearly completely when the water becomes too hot or water quality deteriorates. New methods are needed to greatly increase settlement, growth, survival, and resistance to environmental stress of keystone marine organisms in order to maintain critical coastal ecosystem functions including shore protection, fisheries, and biodiversity. Electrolysis methods have been applied to marine ecosystem restoration since 1976, with spectacular results (Figures 1(a)-(c)). This paper provides the first overall review of the data. Low-voltage direct current trickle charges are found to increase the settlement of corals 25.86 times higher than uncharged control sites, to increase the mean growth rates of reef-building corals, soft corals, oysters, and salt marsh grass— an average of 3.17 times faster than controls (ranging from 2 to 10 times depending on species and conditions), and to increase the survival of electrically charged marine organisms—an average of 3.47 times greater than controls, with the biggest increases under the most severe environmental stresses. These results are caused by the fundamental biophysical stimulation of natural biochemical energy production pathways, used by all organisms, provided by electrical stimulation under the right conditions. This paper reviews for the first time all published results from properly designed, installed, and maintained projects, and contrasts them with those that do not meet these criteria.