Physiological effects of micro-plastics on the red algae, Grateloupia turuturu and Chondrus sp.

Plastics are the most abundant marine litter in the world's oceans and can be degraded into micro-plastics. These emerging pollutants negatively affect marine organisms, but little is known on the effects on macroalgae. In this study, we investigated the effects of micro-plastics on two species of red algae: Grateloupia turuturu and Chondrus sp. Grateloupia turuturu has a slippery surface while Chondrus sp. has a rough surface. Different surface characteristics of these macroalgae may affect adherence of micro-plastics. Both species were exposed to 5 different concentrations (0, 20, 200, 2000 and 20,000 ng/L) of polystyrene microspheres. Adherence capacity, accumulating micro-plastics on the surface was higher for Chondrus sp. than G. turuturu. Chondrus sp. at 20,000 ng/L only showed a decrease in growth rate and photosynthesis activity, and an increase of reactive oxygen species (ROS). However, G. turuturu was not significantly affected by micro-plastics at all tested concentrations. Shaded light and inhibition of gas flow by adhered micro-plastics may be a reason for the reduction of growth and photosynthesis and production of ROS. Based on this result, the toxic effects of micro-plastics appear to be species specific, dependent on the adherence capacity of macroalgae.

A mixed acid treatment for the prevention of Ulva prolifera attachment to Neopyropia aquaculture rafts: Laboratory experimentation.

The sessile green macroalgae in the Neopyropia aquaculture areas of the Subei Shoal are a confirmed source of green tides in the Southern Yellow Sea (SYS) of China. The green tides have a significant impact on local economic development (tourism, aquaculture, etc.) and ecological stability. In order to develop an effective method for eliminating the green macroalgae attached to Neopyropia aquaculture rafts, this study investigated the effects of mixed acid solutions (0.0475 % hydrochloric acid [HCl] and pH 2.0 citric acid) on cell damage, chlorophyll composition, phycobiliprotein content, and the photosynthetic fluorescence characteristics of Ulva prolifera and Neopyropia yezoensis. The chlorophyll contents of U. prolifera and N. yezoensis were significantly affected by the mixed acid solutions. After treatment with a mixed acid solution (4:3 volume ratio of HCl to citric acid) for 5 s, the photosynthetic pigment content of U. prolifera was significantly different from that of normal U. prolifera. However, photosynthetic pigment content in the treated N. yezoensis increased significantly. In addition, mixed acid solution treatment had a significant effect on the Fv/Fm of U. prolifera and N. yezoensis. After mixed acid treatment (4:3 HCl to citric acid), U. prolifera completely died, but the Fv/Fm of N. yezoensis was restored after 3 d. Therefore, a mixed acid solution comprising 0.0475 % HCl and pH 2.0 citric acid (at a volume ratio of 4:3) can be used as an agent for the removal of green macroalgae from Neopyropia aquaculture areas.

Controlling the main source of green tides in the Yellow Sea through the method of biological competition.

Macroalgal blooms have become a serious threat to public health, fisheries, ecosystems, and global economies. Since 2007, in the Yellow Sea, China, Ulva green tides have occurred for 15 consecutive years. However, effective control methods are limited. Ulva prolifera attached to Neopyropia aquaculture rafts are believed to be the main source of blooms, therefore eliminating Ulva from rafts could effectively prevent and control blooms. We investigated this phenomenon and showed that macroalgae germination was significantly inhibited by dried Neopyropia yezoensis at concentrations of 1.2, 2.4, and 4.8 g DW-1. Also, the inhibitory effects of dried N. yezoensis toward U. prolifera gametes at 2.4 and 4.8 g DW-1 were >90% at day 21. N. yezoensis culture filtrates and thalli were also used to determine dose-dependent inhibition effects on U. prolifera gamete germination. Both were potent and significantly inhibited germination at 1.75-7 g FW-1; the inhibitory effect 7 g FW-1 was >90% at day 21. As N. yezoensis thalli exhibited high inhibitory effects in laboratory experiments, we also performed field studies. N. yezoensis on ropes displayed high inhibitory effects on Ulva attachment and growth. Thus N. yezoensis powder, culture filtrates, and thalli displayed strong inhibitory effects on U. prolifera gametes, suggesting N. yezoensis attachment to ropes could be used to control green tides at the source.

Polyopes affinis Suppressed IFN-γ- and TNF-α-Induced Inflammation in Human Keratinocytes via Down-Regulation of the NF-κB and STAT1 Pathways.

Polyopes affinis is a red algal species commonly found on the South coast and near Jeju Island, Korea. This study aimed to determine whether P. affinis extracts can inhibit the pathogenesis of T-helper-2 (Th2)-mediated inflammation in a human keratinocyte cell line of atopic dermatitis (AD). Cells were incubated with 10 ng/mL of interferon gamma (IFN-γ) and 10 ng/mL of tumor necrosis factor-alpha (TNF-α) at various concentrations of PAB (10, 30, and 60 µg/mL) and PAA (100, 500, and 1000 µg/mL) extracts. A gene-ontology (GO)-enrichment analysis revealed that PAB significantly enriched the genes associated with biological processes such as cell adhesion, immune response, inflammation, and chemokine-mediated pathways. PAB suppressed the expression of the secretory proteins and mRNAs that are associated with the thymus and the production of activation-regulated chemokines (TARC/CCL17) and macrophage-derived chemokines (MDC/CCL22). The effect of the extract on mitogen-activated protein kinases (MAPKs) was related to its inhibition of TARC/CCL17 and MDC/CCL22 production by blocking NF-κB and STAT1 activation. These results suggest that seaweed extract may improve AD by regulating pro-inflammatory chemokines. In conclusion, we first confirmed the existence of phloroglucinol, a polyphenol formed from a precursor called phlorotannin, which is present in PAB, and this result proved the possibility of PAB being used as a treatment for AD.

Validation of Direct Boiling Method for Simple and Efficient Genomic DNA Extraction and PCR-based Macroalgal Species Determination.

A rapid, simple, and cost-effective total DNA extraction method referred to as a direct boiling method for PCR-based algal species determination was validated using representative species of green, brown, and red algae. Each sample was briefly minced in two buffers, Tris-EDTA (TE) or PCR buffer, and transferred to a heat block for boiling at 98°C for 5 min. No detergent was used in this experiment. The entire DNA isolation procedure was completed within 10 min. After brief centrifugation, the supernatant was directly used as a template for PCR. As a result, all genomic DNA markers were successfully amplified and sequenced from each algal taxon. Regardless of DNA quality, the direct boiling method is very suitable to identify unknown species of algae from a large amount of samples in a limited time and can be applied broadly to routine seasonal or annual monitoring of algae. DNA from an Undaria pinnatifida sporophyte, however, was not successfully extracted by this direct boiling method, probably due to a high concentration of polysaccharides.

Factors controlling the distributions of dissolved organic matter in the East China Sea during summer.

To determine the distribution of dissolved organic matter (DOM) in the East China Sea (ECS) during the summer, we measured the dissolved organic carbon (DOC) and nitrogen (DON), fluorescent dissolved organic matter (FDOM), and chlorophyll a (Chl. a) in the upper 100-m layer of this region during July and September 2015. The DOC (r2 = 0.72 and 0.78 in July and September, respectively) and DON (r2 = 0.43 and 0.33) were significantly correlated with salinity, suggesting that the river is the primary origin of DOM. However, we found that at a DOC "pulse" under a salinity ranging from 24 to 35, the extrapolating DOC values (304 ± 11 µM) were twice higher than those with a salinity of close to 0, as found in a previous study. The excess DOC concentration seemed to be attributed to the microbial metabolism during transport from the estuary based on the good relationships between DOC and marine humic-like FDOM (r2 = 0.42 and 0.47), as well as the fluorescence, humification, and biological indexes, but showed no correlation with Chl. a. Thus, the results of our study indicate that microbial activities can be a significant factor controlling the distribution of DOM in the ECS during summer.

Microcystin-LR bioconcentration induces antioxidant responses in the digestive gland of two marine bivalves Crassostrea gigas and Mytilus edulis.

Microcystins (MCs) are a major group of potent cyanobacterial toxins found in freshwater and even brackish waterbodies. To understand the putative correlation between bioconcentration of MCs and antioxidant responses of the digestive gland of bivalves, Pacific oyster Crassostrea gigas and blue mussel Mytilus edulis were exposed to different concentrations (0.1, 1, 10 and 20µgL-1) of MC-Leucine-Arginine (LR) for seven days. MC-LR bioconcentrated in the digestive glands of both bivalves during exposure period. The levels were slightly reduced when the bivalves were exposed to seawater during depuration (7days), while approximately 0.1µgL-1 of MC-LR was observed in the 10 and 20µgL-1 exposed bivalves at the end of depuration. Intracellular malondialdehyde (MDA) and glutathione (GSH) levels were significantly elevated in the 10 and 20µgL-1 exposed bivalves at 7day, and the levels were maintained during depuration in both bivalves. Overall, significant higher levels of enzymatic activities of antioxidant defense systems such as glutathione S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GR) were observed in the 10 and 20µgL-1 exposed bivalves. Interestingly, most of higher levels of Pacific oyster were detected at exposure period, while blue mussel showed higher levels at depuration phase, suggesting a species-specific sensitivity upon MC-LR. These patterns were correlated with the bioconcentration patterns of MC-LR as Pacific oyster was highly accumulated by MC-LR during exposure period, but blue mussel showed prolonged high levels of MC-LR for depuration phase. Our results will be useful to understand species-specific bioconcentration of MC-LR in bivalves and their effects on intracellular oxidative status via accumulation.

Effects of Antifouling Biocides on Molecular and Biochemical Defense System in the Gill of the Pacific Oyster Crassostrea gigas.

Antifouling biocides such as organotin compounds and their alternatives are potent toxicants in marine ecosystems. In this study, we employed several molecular and biochemical response systems of the Pacific oyster Crassostrea gigas to understand a potential mode of action of antifouling biocides (i.e. tributyltin (TBT), diuron and irgarol) after exposure to different concentrations (0.01, 0.1, and 1 µg L-1) for 96 h. As a result, all the three antifouling biocides strongly induced the antioxidant defense system. TBT reduced both enzymatic activity and mRNA expression of Na+/K+-ATPase and acetylcholinesterase (AChE). Lower levels of both Na+/K+-ATPase activity and AChE mRNA expression were observed in the diuron-exposed oysters compared to the control, while the irgarol treatment reduced only the transcriptional expression of AChE gene. We also analyzed transcript profile of heat shock protein (Hsp) superfamily in same experimental conditions. All antifouling biocides tested in this study significantly modulated mRNA expression of Hsp superfamily with strong induction of Hsp70 family. Taken together, overall results indicate that representative organotin TBT and alternatives have potential hazardous effects on the gill of C. gigas within relatively short time period. Our results also suggest that analyzing a series of molecular and biochemical parameters can be a way of understanding and uncovering the mode of action of emerging antifouling biocides. In particular, it was revealed that Pacific oysters have different sensitivities depend on the antifouling biocides.

The expansion of Ulva prolifera O.F. Müller macroalgal blooms in the Yellow Sea, PR China, through asexual reproduction.

Since 2007, Ulva macroalgal blooms have occurred along the coastal areas of the Yellow Sea, China. These blooms are dominated by fragments of Ulva prolifera in the early stages of development. The objectives of this study were to identify the primary mode of asexual reproduction for U. prolifera and to evaluate the contribution of these thalli fragments to the formation of blooms. Four different growth and reproductive strategies of U. prolifera segments were found including: 1) tubular diameter becoming larger; 2) formation of new branches; 3) release of zoids; and 4) polarized growth. This is the first report showing the development of numerous blade-lets from a single segment, which is remarkably different from previous studies on other Ulva species. The results in the present study provide critical information to understand how this species is able to support its explosive growth during a bloom.

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.

The origin of the Ulva macroalgal blooms in the Yellow Sea in 2013.

Green algal blooms have occurred in the Yellow Sea for seven consecutive years from 2007 to 2013. In this study, satellite image analysis and field shipboard observations indicated that the Ulva blooms in 2013 originated in the Rudong coast. The spatial distribution of Ulva microscopic propagules in the Southern Yellow Sea also supported that the blooms originated in the Rudong coast. In addition, multi-source satellite data were used to evaluate the biomass of green algae on the Pyropia aquaculture rafts. The results showed that approximately 2784 tons of Ulva prolifera were attached to the rafts and possessed the same internal transcribed spacer and 5S rDNA sequence as the dominant species in the 2013 blooms. We conclude that the significant biomass of Ulva species on the Pyropia rafts during the harvesting season in radial tidal sand ridges played an important role in the rapid development of blooms in the Yellow Sea.