Microbial nitrogen removal in reef-building corals: A light-sensitive process.

Scleractinian corals are the main framework-building groups in tropical coral reefs. In the coral holobiont, nitrogen-cycling mediated by microbes is fundamental for sustaining the coral reef ecosystems. However, little direct evidence characterizing the activities of microbial nitrogen removal via complete denitrification and anaerobic ammonium oxidation (anammox) in stony corals has been presented. In this study, multiple incubation experiments using 15N-tracer were conducted to identify and characterize N2 production by denitrification and anammox in the stony coral Pocillopora damicornis. The rates of denitrification and anammox were recorded up to 0.765 ± 0.162 and 0.078 ± 0.009 nmol N2 cm-2 h-1 respectively. Denitrification contributed the majority (∼90%) of N2 production by microbial nitrogen removal in stony corals. The microbial nitrogen removal activities showed diel rhythms, which might correspond to photosynthetic oxygen production. The N2 production rates of anammox and denitrification increased with incubation time. To the authors' knowledge, this study is the first to confirm and characterize the activities of complete denitrification and anammox in stony corals via stable isotope techniques. This study extends the understanding on nitrogen-cycling in coral reefs and how it participates in corals' resilience to environmental stressors.

Increased light intensity enhances photosynthesis and biochemical components of red macroalga of commercial importance, Kappaphycus alvarezii, in response to ocean acidification.

The concentration of atmospheric carbon dioxide (CO2) has increased drastically over the past several decades, resulting in the pH of the ocean decreasing by 0.44 ± 0.005 units, known as ocean acidification (OA). The Kappaphycus alvarezii (Rhodophyta, Solieriaceae), is a commercially and ecologically important red macroalga with significant CO2 absorption potential from seawater. The K. alvarezii also experienced light variations from self-shading and varied cultivation depths. Thus, the aim of present study was to investigate the effects of two pCO2 levels (450 and 1200 ppmv) and three light intensities (50, 100, and 150 µmol photons·m-2·s-1) on photosynthesis and the biochemical components in K. alvarezii. The results of the present study showed that a light intensity of 50 µmol photons·m-2·s-1 was optimal for K. alvarezii photosynthesis with 0.663 ± 0.030 of Fv/Fm and 0.672 ± 0.025 of Fv'/Fm'. Phycoerythrin contents at two pCO2 levels decreased significantly with an increase in light intensity by 57.14-87.76%, while phycocyanin contents only decreased from 0.0069 ± 0.001 mg g-1 FW to 0.0047 ± 0.001 mg g-1 FW with an increase in light intensity at 1200 ppmv of pCO2. Moreover, moderate increases in light intensity and pCO2 had certain positive effects on the physiological performance of K. alvarezii, specifically in terms of increasing soluble carbohydrate production. Although OA and high light levels promoted total organic carbon accumulation (21.730 ± 0.205% DW) in K. alvarezii, they had a negative impact on total nitrogen accumulation (0.600 ± 0.017% DW).

High nutrient availability modulates photosynthetic performance and biochemical components of the economically important marine macroalga Kappaphycus alvarezii (Rhodophyta) in response to ocean acidification.

Increased atmospheric CO2 concentrations not only change the components of inorganic carbon system in seawater, resulting in ocean acidification, but also lead to decreased seawater pH, resulting in ocean acidification. Consequently, increased inorganic carbon concentrations in seawater provide a sufficient carbon source for macroalgal photosynthesis and growth. Increased domestic sewage and industrial wastewater discharge into coastal areas has led to nutrient accumulation in coastal seawaters. Combined with elevated pCO2 (1200 ppmv), increased nutrient availability always stimulates the growth of non-calcifying macroalgae, such as red economical macroalga Gracilariopsis lemaneiformis. Here, we evaluated the interactive effects of nutrients with elevated pCO2 on the economically important marine macroalga Kappaphycus alvarezii (Rhodophyta) in a factorial 21-day coupling experiment. The effects of increased nutrient availability on photosynthesis and photosynthetic pigments of K. alvarezii were greater than those of pCO2 concentration. The highest Fv/Fm values (0.660 ± 0.019 and 0.666 ± 0.030, respectively) were obtained at 2 µmol L-1 of NO3-N at two pCO2 levels. Under the elevated pCO2 condition, the Chl-a content was lowest (0.007 ± 0.004 mg g-1) at 2 µmol L-1 of NO3-N and highest (0.024 ± 0.002 mg g-1) at 50 µmol L-1 of NO3-N. The phycocyanin content was highest (0.052 ± 0.012 mg g-1) at 150 µmol L-1 of NO3-N under elevated pCO2 condition. The malondialdehyde content declined from 32.025 ± 4.558 nmol g-1 to 26.660 ± 3.124 nmol g-1 with the increased nutrients at under low pCO2. To modulate suitable adjustments, soluble biochemical components such as soluble carbohydrate, soluble protein, free amino acids, and proline were abundantly secreted and were likely to protect the integrity of cellular structures under elevated nutrient availability. Our findings can serve as a reference for cultivation and bioremediation methods under future environmental conditions.

Toxicological effects of oxybenzone on the growth and bacterial composition of Symbiodiniaceae.

Oxybenzone, a common ultraviolet (UV) filter, is a growing environmental concern due to its ecotoxicological effects. However, the responses of Symbiodiniaceae and their bacterial communities to oxybenzone are largely unknown. In this study, the effects of oxybenzone on Effrenium voratum and Cladocopium goreaui were investigated. The results revealed that sensitivity of Symbiodiniaceae to oxybenzone was species-dependent. 50 µg L-1 of oxybenzone significantly impacted the cell density of C. goreaui, causing a 36.73% decrease. When oxybenzone concentration increased to 500 µg L-1 and 5000 µg L-1, cell division was completely suppressed; meanwhile, chl-a content declined to zero. Compared to C. goreaui, E. voratum had higher resistance to oxybenzone. There was no significant difference in cell density between 50 µg L-1 group and control group. At higher dosage of oxybenzone (500 µg L-1 and 5000 µg L-1), the cell density declined 32.02% and 45.45% compared to the control group, respectively. Additionally, we revealed that the diversity and structure of bacterial community were affected by oxybenzone. Briefly, 500 µg L-1 and 5000 µg L-1 of oxybenzone altered the diversity of bacterial community in C. goreau. Furthermore, the relative abundances of Costertonia, Roseitalea, Rhodopirellula, and Roseobacter were negatively affected by oxybenzone ranging 50 µg L-1 to 5000 µg L-1. Compare to C. goreaui, the bacterial community composition associated with E. voratum was more stable. As revealed by KEGG pathway analysis, oxybenzone affected energy metabolism and inhibited the metabolism of cofactors and vitamins in C. goreaui, while 5000 µg L-1 of oxybenzone significantly altered the carbohydrate metabolism, membrane transport and amino acid metabolism in E. voratum. The changes of bacterial composition may contribute to the variation in algal growth. These results indicated that oxybenzone pollution could injury Symbiodiniaceae, even threaten coral reef ecosystems.

Response mechanisms to ocean warming exposure in Effrenium voratum (Symbiodiniaceae).

Ocean warming is an extreme environment event that has profound and lasting impacts on Symbiodiniaceae. However, their response mechanisms to elevated temperature exposure are poorly understood. In this study, the physiological and transcriptional responses of Effrenium voratum (Symbiodiniaceae) to ocean warming were examined. After exposure to 30 °C, no significant variations in growth, chlorophyll a, or photosynthetic and respiration rates were observed, while a higher temperature (34 °C) significantly reduced these physiological measurements. Meanwhile, lipid content and fatty acid composition were altered at high temperature (i.e., elevated degree of fatty acid saturation). Such biochemical constituents likely contributed to the mitigation of the negative effects of elevated temperatures. Furthermore, higher expression levels of genes related to the synthesis and elongation of fatty acids were detected at high temperature. The adjustment of lipids and fatty acid composition may be a potential mechanism by which E. voratum may survive under future global warming. ONE SENTENCE SUMMARY: The adjustment of lipids and fatty acid composition may be a potential mechanism by which E. voratum acclimate to future global warming.

Biochar Addition Altered Bacterial Community and Improved Photosynthetic Rate of Seagrass: A Mesocosm Study of Seagrass Thalassia hemprichii.

Seagrass meadows, as typical "blue carbon" ecosystems, play critical ecological roles in the marine ecosystem and decline every year. The application of biochar in soil has been proposed as a potential soil amendment to improve soil quality and mitigate global climate change. The effects of biochar on soil bacterial activities are integrally linked to the potential of biochar in achieving these benefits. However, biochar has been rarely applied in marine ecosystems. Whether the application of biochar could work on the seagrass ecosystem remained unknown. In this study, we investigated the responses of sediment and rhizosphere bacterial communities of seagrass Thalassia hemprichii to the biochar addition derived from maize at ratios of 5% by dry weight in the soil during a one-month incubation. Results indicated that the biochar addition significantly changed the sedimental environment with increasing pH, total phosphorus, and total kalium while total nitrogen decreased. Biochar addition significantly altered both the rhizosphere and sediment bacterial community compositions. The significant changes in rhizosphere bacterial community composition occurred after 30days of incubation, while the significant variations in sediment bacterial community composition distinctly delayed than in sediment occurred on the 14th day. Biochar application improved nitrification and denitrification, which may accelerate nitrogen cycling. As a stabilizer to communities, biochar addition decreased the importance of deterministic selection in sediment and changed the bacterial co-occurrence pattern. The biochar addition may promote seagrass photosynthesis and growth by altering the bacterial community compositions and improving nutrient circulation in the seagrass ecosystem, contributing to the seagrass health improvement. This study provided a theoretical basis for applying biochar to the seagrass ecosystem and shed light on the feasible application of biochar in the marine ecosystem. Graphical Abstract.

Bacterial Communities Associated With Healthy and Bleached Crustose Coralline Alga Porolithon onkodes.

Crustose coralline algae (CCA) play vital roles in producing and stabilizing reef structures and inducing the settlement and metamorphosis of invertebrate larvae in coral reef ecosystems. However, little is known about the bacterial communities associated with healthy and bleached CCA and their interactions with coral larval settlement. We collected samples of healthy, middle semi-bleached, and bleached CCA Porolithon onkodes from Sanya Bay in the South China Sea and investigated their influences on the larval settlement and metamorphosis of the reef-building coral Pocillopora damicornis. The larval settlement/metamorphosis rates all exceeded 70% when exposed to healthy, middle semi-bleached, and bleached algae. Furthermore, the compositions of bacterial community using amplicon pyrosequencing of the V3-V4 region of 16S rRNA were investigated. There were no obvious changes in bacterial community structure among healthy, middle semi-bleached, and bleached algae. Alphaproteobacteria, Bacteroidetes, and Gammaproteobacteria were dominant in all samples, which may contribute to coral larval settlement. However, the relative abundances of several bacterial communities varied among groups. The relative abundances of Mesoflavibacter, Ruegeria, Nautella, and Alteromonas in bleached samples were more than double those in the healthy samples, whereas Fodinicurvata and unclassified Rhodobacteraceae were significantly lower in the bleached samples. Additionally, others at the genus level increased significantly from 8.5% in the healthy samples to 22.93% in the bleached samples, which may be related to algal bleaching. These results revealed that the microbial community structure associated with P. onkodes generally displayed a degree of stability. Furthermore, bleached alga was still able to induce larval settlement and metamorphosis.

Transcriptomic and Physiological Responses of the Tropical Reef Calcified Macroalga Amphiroa fragilissima to Elevated Temperature1.

Calcareous macroalgae are of particular ecological importance as primary producers, carbonate sediment builders, and habitat providers in coral reef ecosystems. Ocean warming is a major threat to calcareous algae, but it remains unclear exactly how these algae will respond to it. In this study, the potential physiological impacts of ocean warming on the calcareous alga Amphiroa fragilissima were evaluated in laboratory experiments. Increasing temperature from 26 to 28°C had positive effects on algal growth rate and chlorophyll a content, but these parameters decreased significantly at 32°C, which is 5°C above the annual mean temperature in the study region. Algal bleaching occurred at 34°C. There were no significant differences in CaCO3 content of thalli among different temperatures; however, calcification rate was inhibited significantly at 32 and 34°C. Transcriptome analyses using the Illumina RNA-seq platform showed that differentially expressed genes were annotated mainly in the categories of steroid biosynthesis, gap junction, ribosome, and mTOR signaling pathway. The expression levels of PsbA and PsbP were suppressed at 32°C, implying that inactivation of photosystem II could be a main reason for the decreased photosynthetic rate. Down-regulation of the genes encoding carbonic anhydrase and nitrate reductase was observed at 32°C, which could inhibit growth rate. Additionally, several genes that might be related to calcification were identified, including CAMK, CDPK, and CAM and genes encoding alpha-catenin and carbonic anhydrase. This study contributes to our understanding of the effects of temperature on algal calcification and provides a theoretical basis to protect ecological diversity of coral reef ecosystems.

Calcified macroalgae and their bacterial community in relation to larval settlement and metamorphosis of reef-building coral Pocillopora damicornis.

Calcified macroalgae play an important role in the settlement and metamorphosis of invertebrate larvae in coral reef ecosystems. However, little is known about the algal-associated bacterial communities and their effects on larval settlement. In this study, the responses of larvae of the coral Pocillopora damicornis to calcified algae (Porolithon onkodes, Halimeda cylindracea, Halimeda opuntia and Amphiroa fragilissima) were evaluated. The results revealed that Por. onkodes and H. cylindracea significantly enhanced the rates of settlement and metamorphosis, whereas fewer larvae settled on Am. fragilissima and H. opuntia. Amplicon pyrosequencing of the V3-V4 region of 16S rDNA was applied to investigate the relationship between algal bacterial community and larval settlement. Principal coordinates analysis demonstrated that the bacterial community composition of H. opuntia was more similar to that of Am. fragilissima, but clearly distinct from those of H. cylindracea and Por. onkodes. Furthermore, the relative abundances of bacteria were highly diverse among different algae. H. opuntia had higher percentages of Thalassobius, Pelagibius and SM1A02, whereas the abundances of Mycoplasma and Suttonella were significantly higher in H. cylindracea than other algae. Our results showed that larval settlement/metamorphosis was strongly correlated with the bacterial community composition and with the relative abundance of a few operational taxonomic units.

Optimization of medium using response surface methodology to enhance the growth of Effrenium voratum (Symbiodiniaceae, Dinophyceae).

Survival of coral reef-associated Symbiodiniaceae is vital to maintain the healthy coral community in coral reefs. However, knowledge about cultivation of free-living or symbiotic Symbiodiniaceae has been limited. In this study, the response surface methodology was applied to optimize the medium for Effrenium voratum. The results showed that the impacts of nutrient components on algal growth were: FeCl3 > NaH2 PO4 >MnSO4 > MgSO4 /CoSO4 > KCl>ZnSO4 > CaCl2 /NaNO3 , among which NaH2 PO4 and FeCl3 significantly affected algal growth. The optimal medium was: natural seawater supplemented with NaH2 PO4 ·2H2 O 0.25 mM,FeCl3 ·6H2 O 14.24 µM, NaNO3 0.94 mM, MgSO4 ·7H2 O 40.63 mM, KCl 5.37 mM, CaCl2 ·2H2 O 4.08 mM, ZnSO4 ·7H2 O 0.35 µM, MnSO4 9.93 µM, and CoSO4 0.36 µM. The use of the optimized medium resulted in an increase of biomass yield (0.76 g dry weight · L-1 ) by 46% over that using the initial medium, which agreed with the predicted value (0.71 g · L-1 ). Additionally, fatty acids, mainly consisting of palmitic acid (C16:0) and ethyl carbonate (C20:0), accounted for approximately 50% of the total fatty acids in E. voratum. Interestingly, docosahexaenoic acid (DHA) accounted for 6% of total fatty acids, a high proportion that makes E. voratum a potential candidate feedstock in aquaculture for DHA production.

[Pathogenicity of white-spot syndrome virus in Macrobrachium nipponensis via different infection routes].

Macrobrachium nipponensis is delicious and has high economic value, but its susceptibility to white-spot syndrome virus (WSSV) is unknown. Susceptibility, morbidity, and multiplication of WSSV in M. nipponense were studied by epidemiological survey, infection experiment and qPCR. M. nipponense was the natural host of WSSV, and the natural carrying rate was about 8.33%. M. nipponense could be infected with WSSV via oral administration, muscle injection and immersion, and the cumulative infection rate of 10 d exposure was 100%, and the cumulative mortality rates were 100%, 75% and 0%, respectively. The infection of WSSV is fast by muscle injection. The virus content after 5 day's injection is 1 000 times higher than that of the first day of infection, and the mortality rate reached 100% after 8 days. The median lethal dose (LD50) measured as the mortality of infected M. nipponense via injection indicated the LD50 in the concentration of WSSV of 2.71×105 virions/µL. In shrimp farming, M. nipponense can be infected by ingesting WSSV infected shrimp or dead shrimp, and also by soaking in WSSV-containing water and thus become a vector, consequently affecting the spread and pathogenicity of WSSV.

Bioremediation using Gracilaria lemaneiformis to manage the nitrogen and phosphorous balance in an integrated multi-trophic aquaculture system in Yantian Bay, China.

To reduce negative environmental impacts from human aquaculture activities, the red alga Gracilaria lemaneiformis was co-cultured with the fish Pseudosciaena crocea in an integrated multi-trophic aquaculture (IMTA) system for 35d in Yantian Bay. The eutrophication index value decreased from 14.5 to 8.4 after seaweeds were co-cultured in cage farming areas, which indicated that the eutrophic water column in Yantian Bay could be mediated by IMTA. Total DIN and DIP of the tidal input and output were 9.23kg, 0.19kg and 11.08kg, and 0.27kg, respectively. Total 5.24kg of dissolved N and 0.81kg of dissolved P were released from IMTA system. These results indicate that G. lemaneiformis co-cultured in IMTA system could not completely remove all excess nutrients. In theory, at least 324.48kg of seaweed seedlings would be required to balance excess nutrients generated from fish cages.

Tracing the origin of green macroalgal blooms based on the large scale spatio-temporal distribution of Ulva microscopic propagules and settled mature Ulva vegetative thalli in coastal regions of the Yellow Sea, China.

From 2008 to 2016, massive floating green macroalgal blooms occurred annually during the summer months in the Yellow Sea. The original source of these blooms was traced based on the spatio-temporal distribution and species composition of Ulva microscopic propagules and settled Ulva vegetative thalli monthly from December 2012 to May 2013 in the Yellow Sea. High quantities of Ulva microscopic propagules in both the water column and sediments were found in the Pyropia aquaculture area along the Jiangsu coast before a green macroalgal bloom appeared in the Yellow Sea. The abundance of Ulva microscopic propagules was significantly lower in outer areas compared to in Pyropia aquaculture areas. A molecular phylogenetic analysis suggested that Ulva prolifera microscopic propagules were the dominant microscopic propagules present during the study period. The extremely low biomass of settled Ulva vegetative thalli along the coast indicated that somatic cells of settled Ulva vegetative thalli did not provide a propagule bank for the green macroalgal blooms in the Yellow Sea. The results of this study provide further supporting evidence that the floating green macroalgal blooms originate from green macroalgae attached to Pyropia aquaculture rafts along the Jiangsu coastline of the southern Yellow Sea.

Eutrophication assessment and bioremediation strategy using seaweeds co-cultured with aquatic animals in an enclosed bay in China.

Intensive mariculture results in a rise in nutrient concentrations, then leads to serious eutrophication in coastal waters. Based on the sampling data obtained between August 2012 and July 2013, the eutrophication status in Yantian Bay was assessed, and the proportion of marine animals co-cultured with seaweeds was evaluated. The nutritional quality index (NQI) ranged from 4.37 to 13.20, indicating serious eutrophication conditions. The annual average ratio of nitrogen/phosphorus (N/P) was 25.19, indicating a nitrogen surplus in this system. DIN was selected as the best parameter to balance seaweed absorption and marine animal DIN production. Gracilaria lemaneiformis and Laminaria japonica were selected as co-cultured seaweeds. The optimal proportion of G. lemaneiformis production was assessed as 20074.14 tonnes. The optimal proportion of L. japonica production was evaluated as 15890.68 tonnes. High-temperature adapted seaweeds should be introduced for removing nutrients releasing by farmed aquatic animals in the summer in Yantian Bay.