In the current study, the toxicological mechanisms of microcystin-LR and its disadvantageous effects on Daphnia magna were examined. Survival rate, number of newborn, activity of several important enzymes [glutathione S-transferase (GST), lactate dehydrogenase (LDH), phosphatases, and glutathione], accumulated microcystins, and ultrastructural changes in different organs of Daphnia were monitored over the course of 21-day chronic tests. The results indicated that low concentrations of dissolved microcystin had no harmful effect on Daphnia. On the contrary, stimulatory effects were detected. In the presence of toxin at high dosage and for long-term exposure, GST and glutathione levels decreased significantly. The decreased enzyme activity in the antioxidant system probably was caused by detoxification reactions with toxins. And these processes of detoxification at the beginning of chronic tests may enable phosphatases in Daphnia magna to withstand inhibition by the toxins. At the same time, we also found that the LDH activity in test animals increased with exposure to microcystin-LR, indicating that adverse effects occurred in Daphnia. With microcystin given at a higher dosage or for a longer exposure, the effect on Daphnia magna was fatal. In the meantime, microcystin began to accumulate in Daphnia magna, and phosphatase activity started to be inhibited. From the ultrastructure results of cells in D. magna, we obtained new information: the alimentary canal may be the target organ affected by exposure of microcystins to D. magna. The results of the current study also suggested that the oxidative damage and PPI (protein phosphatase inhibition) mechanisms of vertebrates also are adapted to Daphnia.

A Chrysophyceae species, Poterioochromonas sp., was isolated from Microcystis cultures. This species can efficiently prey on Microcystis and can grow faster phagotrophically than autotrophically. The growth of Poterioochromonas sp. was stimulated in the presence of microcystin-LR and microcystin-RR (in concentrations ranging from 0.1 to 4 mg/L). The growth rate of Poterioochromonas was 4-5 times higher than the control, indicating the toxins serve as growth stimuli for this organism. A subculture of toxin-treated cells, however, showed low cellular viability, suggesting that growth enhancement by microcystins was not a normal process. The antioxidant enzymatic activity of Poterioochromonas sp. was screened for toxicology analysis. Glutathione, malondialdehyde, and superoxide dismutase (SOD) content was up-regulated within 8 h of exposure to microcystin-LR (500 microg/L). A high level of SOD activity during exposure to the toxin indicated that SOD was involved in decreasing oxidative stress caused by microcystin-LR. Simultaneously with growth, Poterioochromonas was able to degrade microcystin-LR even, at a toxin concentration of 4 mg/L. This putative degradation mechanism in Poterioochromonas is explored further and discussed in this article. Our findings may shed light on understanding the role of Poterioochromonas in the aquatic ecosystem, in particular, as a grazer of toxic cyanobacteria and a biodegrader for microcystins.

Surface water samples from downstream and estuarine areas of Jiulong River were collected in August 2011 and May 2012 for detecting sulfonamide antibiotic residues and isolating sulfamethazine-resistant bacteria. Sulfamethazine was detected in all samples in May 2012 at an average concentration of 78.3 ng L−1, which was the highest among the nine sulfonamide antibiotics determined. Sulfamethazine-resistant bacteria (SRB) were screened using antibiotic-containing agar plates. The SRB average abundance in the samples was 3.69 × 104 and 2.17 × 103 CFUs mL−1 in August 2011 and May 2012, respectively, and was positively correlated to sulfamethazine concentration in May 2012. The 16S rRNA gene sequencing of all the 121 SRB isolates revealed high diversity. Furthermore, the SRB isolates exhibited multidrug resistance, with 48.7 % showing resistance to at least three antibiotics. The abundance and persistence of highly diverse SRB and their multidrug resistance are likely to demonstrate the transferable pressure from coastal environments on public health.Electronic supplementary materialThe online version of this article (doi:10.1007/s11356-015-4473-z) contains supplementary material, which is available to authorized users.

Bacteria play a pivotal role in shaping ecosystems and contributing to elemental cycling and energy flow in the oceans. However, few studies have focused on bacteria at a trans-basin scale, and studies across the subtropical Northwest Pacific Ocean (NWPO), one of the largest biomes on Earth, have been especially lacking. Although the recently developed high-throughput quantitative sequencing methodology can simultaneously provide information on relative abundance, quantitative abundance, and taxonomic affiliations, it has not been thoroughly evaluated. We collected surface seawater samples for high-throughput, quantitative sequencing of 16S rRNA genes on a transect across the subtropical NWPO to elucidate the distribution of bacterial taxa, patterns of their community structure, and the factors that are potentially important regulators of that structure. We used the quantitative and relative abundances of bacterial taxa to test hypotheses related to their ecology. Total 16S rRNA gene copies ranged from 1.86 × 108 to 1.14 × 109 copies L−1. Bacterial communities were distributed in distinct geographical patterns with spatially adjacent stations clustered together. Spatial considerations may be more important determinants of bacterial community structures than measured environmental variables. The quantitative and relative abundances of bacterial communities exhibited similar distribution patterns and potentially important determinants at the whole-community level, but inner-community connections and correlations with variables differed at subgroup levels. This study advanced understanding of the community structure and distribution patterns of marine bacteria as well as some potentially important determinants thereof in a subtropical oligotrophic ocean system. Results highlighted the importance of considering both the quantitative and relative abundances of members of marine bacterial communities.

SUMMARYThe fluorescein diacetate (FDA) assay has been widely used to measure metabolic activity in phytoplankton. It was found that FDA fluorescence values did not decrease in some stressed cells, demonstrating that the applicability of the method needs to be assessed further in the context of growth-influencing conditions. In the present study, changes of FDA fluorescence values were studied in bloom-forming cyanobacterial Microcystis aeruginosa Kütz cells under stress conditions such as nitrogen (N) or phosphorus (P) deficiency, or darkness and low temperature (10°C), respectively. The results demonstrated that esterase activity decreased immediately in dark-stressed cells, which correlated with the decline of biomass and photosynthetic activity. Under the other three stress conditions, however, especially at low temperature, the cells lost photosynthetic activity but had the highest esterase activity, which was five times higher than the control group. These findings contrast with the assay criteria that the expression of a stain should reflect the change of photosynthetic activity and that stressed cells should have a lower staining intensity than the control cells. According to these results, the esterase activity response was dependent on environmental factors. Furthermore, higher fluorescence intensity did not mean higher metabolic activity, but a discrepant value indicated a severe stress.

Our study examined the relationship of microphytobenthos to greenhouse gas fluxes from sediments of a subtropical mangrove forest and adjacent mudflat in the Jiulong River Estuary, South China. The relationship between chlorophyll a concentration at the sediment surface and diatom density confirmed that these microalgae were the important component of the microphytobenthos, which produced an observable biofilm in cold seasons (winter and spring) on both the mangrove and mudflat sediment surfaces. Fluxes of methane and nitrous oxide were not affected by the microalgae film and were similar between the mangrove and mudflat. However, benthic microalgae affected the sediment to atmosphere carbon dioxide (CO 2 ) flux, and the effect was temporally variable with the seasonal change in microalgae abundance. In the cold seasons, the mangrove sediment was a CO 2 sink under light chambers but a source under dark chambers. In summer, when there was no visible microalgae film at the sediment surface, the intertidal sediments had CO 2 emissions and comparable fluxes between the two chambers. The negative daily CO 2 fluxes of the film-covered sediment (as the average of the dark and light fluxes) and positive flux of the sediment without visible biofilm indicated that the occurrence of microalgae film converted the mangrove sediment from a CO 2 source to a sink and that the photosynthesis of the microalgae film offset the sediment respiration during the cold seasons in this study. We also found similar effects of microalgae on CO 2 fluxes on the nonvegetated mudflat.

The quantities of greenhouse gas emissions and the activity of functional microbes in coastal soils receiving nutrient-rich wastewaters from mariculture activities have seldom been reported. We investigated the effects of wastewater discharge resulting from dredging shrimp pond sediment on the soil fluxes of methane (CH4) and nitrous oxide (N2O) in intertidal areas and on the functional microorganisms and physio-chemical characteristics of soil. The temporal variations in gas fluxes and soil characteristics following wastewater discharge were also evaluated with the tidal regime on the day of discharge taken into account. The results showed that wastewater discharge immediately resulted in higher levels of ammonia (NH4 +-N) deposited and N2O emissions from the soil at the discharge site than at the non-discharge site, while the CH4 flux was not affected. The increase in N2O flux lasted for a longer time when the discharge was performed during a neap tide day than when it was performed during a spring tide day. Wastewater discharge also increased the abundance of ammonia-oxidizing bacterial (AOB) amoA genes and nosZ genes in soil rather than increasing the abundance of narG and nirK genes. The pattern of temporal variations between the N2O flux and soil NH4 +–N content was similar to that between the flux and the AOB-amoA gene abundance, suggesting that bacterial nitrification was important for N2O production in soil receiving the dredging wastewater. The results suggest that the wastewater discharge impacts nitrogen metabolism processes and causes a significant N2O emission problem; therefore, pollutant management is essential in shrimp culturing activities to reduce greenhouse gas emissions into the atmosphere.

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