Temperature-mediated microbial carbon utilization in China's lakes.

Microbes play an important role in aquatic carbon cycling but we have a limited understanding of their functional responses to changes in temperature across large geographic areas. Here, we explored how microbial communities utilized different carbon substrates and the underlying ecological mechanisms along a space-for-time substitution temperature gradient of future climate change. The gradient included 47 lakes from five major lake regions in China spanning a difference of nearly 15°C in mean annual temperatures (MAT). Our results indicated that lakes from warmer regions generally had lower values of variables related to carbon concentrations and greater carbon utilization than those from colder regions. The greater utilization of carbon substrates under higher temperatures could be attributed to changes in bacterial community composition, with a greater abundance of Cyanobacteria and Actinobacteriota and less Proteobacteria in warmer lake regions. We also found that the core species in microbial networks changed with increasing temperature, from Hydrogenophaga and Rhodobacteraceae, which inhibited the utilization of amino acids and carbohydrates, to the CL500-29-marine-group, which promoted the utilization of all almost carbon substrates. Overall, our findings suggest that temperature can mediate aquatic carbon utilization by changing the interactions between bacteria and individual carbon substrates, and the discovery of core species that affect carbon utilization provides insight into potential carbon sequestration within inland water bodies under future climate warming.

Treatment of coracoid process fractures combined with acromioclavicular joint dislocation using clavicular hook plate.

BACKGROUND: The injury mechanism of acromioclavicular (AC) dislocation combined with coracoid process (CP) fracture is not clear, and there is no consensus on its treatment. This study was performed to evaluate the diagnosis of CP fractures combined with AC dislocation and the effectiveness of operative treatment using a clavicular hook plate. METHODS: Eighteen patients with CP fractures combined with AC dislocation were treated with a clavicular hook plate from May 2012 to June 2021. The patients comprised 10 male and 8 female patients with an average age of 38 years (range, 16-54 years). The injury was caused by falling in 15 patients, traffic accidents in 2 patients, and falling from a height in 1 patient. The Eyres type of CP fracture was type II in 1 patient, type III in 11 patients, type IV in 3 patients, and type V in 3 patients. The Ogawa type of CP fracture was type I in 17 patients and type II in 1 patient. The Rockwood type of AC dislocation was type V in 1 patient, variation type III in 15 patients, and variation type V in 2 patients. The interval from injury to the operation was 3 days (range, 1-7 days). Postoperative complications and CP fracture healing were recorded. Functional assessment at the last follow-up was performed by an independent reviewer using the Constant score and visual analog scale score. RESULTS: All 18 patients were followed up for a mean period of 49 months (range, 12-123 months). Nine patients had acromion osteolysis and 3 patients had CP fracture nonunion (Eyres type II, III, and V in 1 patient each); however, no patients developed shoulder pain, incision infection, limitation of shoulder movement, clinical symptoms of subcoracoid impingement, or AC dislocation relapse. At the last follow-up, the mean Constant score was 99 (range, 94-100). CONCLUSIONS: The possibility of CP fracture should be considered in patients with AC dislocation to avoid a missed diagnosis. Fixation with a clavicular hook plate is a feasible treatment for CP fracture combined with AC dislocation and provides a satisfactory outcome. CP fracture healing may be related to the fracture morphology.

Effects of attapulgite on the growth status of submerged macrophytes Vallisneria spiralis and sediment microenvironment.

The effects of raw attapulgite clay and thermally modified attapulgite clay on the growth status of submerged plant Vallisneria Spiralis (V. spiralis) and the microenvironment of sediment were first explored. The results demonstrated that the attapulgite could effectively promote the development of V. spiralis and improve plant stress resistance by enhancing the activity of antioxidant enzymes. The 10% addition of attapulgite clay increased the biomass of V. spiralis by 27%∼174%, and the promoted rate of raw attapulgite clay was 2∼5 times of modified attapulgite clay. The attapulgite increased redox potential in sediment (P < 0.05) and provided proper niches for organism propagation, further promoting the degradation of organic matter and nutrient metabolism in sediment. The value of Shannon, Chao, and Ace was 9.98, 4865.15, 5029.08 in the 10% modified attapulgite group, and 10.12, 4856.85, 4947.78 in the 20% raw attapulgite group, respectively, indicating that the attapulgite could increase the microbial diversity and abundance in sediment. Additionally, the nutrient elements, such as Ca, Na, S, Mg, K, Zn, and Mo, that dissolved from attapulgite may also promote the V. spiralis growth. This study provided an environment-friendly approach to facilitating submerged macrophyte restoration in the eutrophic lake ecosystem.

Study on adsorption and recovery utilization of phosphorus using alkali melting-hydrothermal treated oil-based drilling cutting ash.

Oil-based drill cutting ash (OBDCA) was treated by alkali melting-hydrothermal method and used as novel adsorbent (AM-HT-OBDCA) for the recovery of phosphorus (P) in water body. The experiment parameter for preparation of AM-HT-OBDCA was optimized, including alkali melting ratio (MOBDCA: MNaOH), alkali melting temperature and hydrothermal temperature. The adsorption process of phosphorus on AM-HT-OBDCA was fit well with the pseudo-second-order model and the Langmuir model. The calculated theoretic adsorption capacity of phosphorus on AM-HT-OBDCA was 62.9 mg/g. The adsorption behavior was spontaneous and endothermic. The effect of pH value and interfering ions on the adsorption of phosphorus in AM-HT-OBDCA was investigated. The main existing form of adsorbed phosphorus on AM-HT-OBDCA was sodium hydroxide extraction form phosphorus (NaOH-P), including iron form phosphorus (Fe-P) and aluminum form phosphorus (Al-P). Precipitation and ligand exchange were the main mechanisms of phosphorus adsorption on AM-HT-OBDCA. The AM-HT-OBDCA used for phosphorus adsorption (AM-HT-OBDCA-P) could be further utilized as fertilizer to promote plant growth. The results of this study provide fundamental data and evaluation support for resource utilization of OBDCA. These results will also provide a reference for the adsorption and recovery utilization of phosphorus using solid waste-based adsorbent.

Potential effects of Cladophora oligoclora Decomposition: Microhabitat variation and Microcystis aeruginosa growth response.

Excessive proliferation of filamentous green algae (FGA) is a new ecological problem in lake systems that have not yet reached a steady state. However, knowledge on how FGA decomposition affects the physical and chemical properties of microhabitats, and whether FGA decomposition stimulates the growth of harmful microalgae in the same niche and promotes the formation of harmful algal blooms remains unclear. In this study, we investigated the decomposing effect of a typical FGA, Cladophora oligoclora, on the density and photosynthetic capacity of Microcystis aeruginosa. C. oligoclora decomposition was characterized under different conditions, namely, unshaded and aerobic, unshaded and anoxic, shaded and anaerobic, and shaded and anoxic, which represented different environmental states in the sedimentation process of decaying C. oligoclora mats from water surface to sediment. The shaded and anaerobic treatment significantly decreased the dissolved oxygen and pH of the culture medium by 66.48 % and 7.21 %, respectively, whereas the conductivity and total organic carbon increased by 71.17 and 70.19 times compared with the control group, respectively. This indicated that the decomposing C. oligoclora deposited at the bottom under dark and anaerobic conditions in natural waters had the greatest impact on the lake environment. Further, the cell density of M. aeruginosa was higher than that in the control group with low concentration (10 % of decomposing solution), whereas the cell density and photosynthetic activity decreased significantly at high concentration of the decomposing solution. Fatty acids and phenolic acids were identified as the main Cyanobacteria-inhibiting active substances in the organic acid components of the decomposing solution. Furthermore, phenol, 4-methyl- and indole compounds were active organic lipophilic compounds in the residue and solution of decomposing C. oligoclora were difficult to degrade. Our findings will be valuable for understanding the succession relationships between FGA and cyanobacteria, which have the same niche in lake ecosystems.

Effect of submerged plant coverage on phytoplankton community dynamics and photosynthetic activity in situ.

Restoration of submerged plants in eutrophic lakes can reduce nutrients and phytoplankton biomass in the water body. However, the effect of submerged plants on phytoplankton communities and their photosynthetic activity in situ are still poorly understood. Here, we studied the response of phytoplankton community structure and fluorescence parameters to different submerged plants coverage, the relationship of phytoplankton community and fluorescence parameters with submerged plants coverage and water physicochemical parameters were analysed in sampling area of Hangzhou West Lakes. The results showed that the coverage and biomass of submerged plants were negatively correlated with nitrogen and phosphorus contents in water body but positively correlated with total phenol content. The ratio of nitrogen to phosphorus in the study site changed greatly (32.25-124.54). In spring and summer, Oscillatoria and Leptolyngbya (Cyanophyta) were the dominant species, while in autumn and winter, the dominant species were Cyclotella (Chlorophyta), and Melosira and Cymbella (Bacillariophyta). Compared with Chlorophyta and Bacillariophyta, fluorescence parameters of Cyanophyta were more sensitive to total phosphorus, N:P ratio, total phenols, pH, and electric conductivity. Fluorescence parameters of Chlorophyta and Bacillariophyta were only affected by underwater light. Total phosphorus (TP) and N:P had a negative effect on the maximum photochemical electron yield of Cyanophyta. Furthermore, Cyanophyta was inhibited by total phenols from submerged plants. When phytoplankton were under stress, photochemical electron yield decreased significantly, whereas non-photochemical quenching increased. The structural equation model showed that the coverage of submerged plants might indirectly affect the fluorescence parameters of Cyanophyta by affecting nitrogen, phosphorus, and total phenol contents in the water body. These findings contribute to the understanding of the mechanisms underlying the impact of submerged plant restoration on phytoplankton community dynamics in subtropical eutrophic shallow lakes and provide a theoretical basis for the management of lakes.

Effects of vermiculite on the growth process of submerged macrophyte Vallisneria spiralis and sediment microecological environment.

Ecological restoration is one of the hot technologies for the reconstruction of eutrophic lake ecosystems in which the restoration and propagation of submerged plants is the key and difficult step. In this paper, the effect of vermiculite on the growth process of Vallisneria spiralis and sediment microenvironment were investigated, aiming to provide a theoretical basis for the application of vermiculite in aquatic ecological restoration. Results of growth indexes demonstrated that 5% and 10% vermiculite treatment groups statistically promote the growth of Vallisneria spiralis compared to the control. Meanwhile, the results of ecophysiological indexes showed that photosynthetic pigment, soluble sugar content, superoxide dismutase (SOD), and catalase (CAT) activity of 5% and 10% group were increased compared with the control while the malondialdehyde (MDA) content exhibited the opposite result (p < 0.05), which illustrated that vermiculite can improve the resistance of plants and delay the aging process of Vallisneria spiralis. In addition, result of PCA (Principal Component Analysis) demonstrated 5% and 10% group has improved the sediment physical conditions and create more ecological niche for microorganisms directly, and then promoted the growth of plants. The dissolution results showed that vermiculite can dissolve the constant and trace elements needed for plant growth. Furthermore, the addition of vermiculite increased the diversity of microorganisms in the sediments, and promoted the increase of plant growth-promoting bacteria and phosphorus-degrading bacteria. This study could provide a technique reference for the further application of vermiculite in the field of ecological restoration.

Etiological characteristics of "tail blister disease" of Australian redclaw crayfish (Cherax quadricarinatus).

In November 2019, an acute disease outbreak in Australian redclaw crayfish (Cherax quadricarinatus) occurred in a farm in Hubei, China, with a cumulative mortality rate of over 80%. One of the characteristic symptoms of the disease was blisters on the tail. This symptom is also common in diseased Procambarus clarkii every year in this country, but the causative agent has not been determined. This study analyzed the etiological characteristics of this disease. Bacterial isolation and identification combined with high-throughput sequencing analysis were conducted to obtain the microbiota characteristics in the hemolymph, hepatopancreas, and intestines. Results showed that this outbreak was caused by infection from Aeromonas hydrophila and Aeromonas veronii. The underlying cause was stress imposed on crayfish during transferring from outdoor pond to indoor pond because of temperature drops. Aeromonas infection caused remarkable changes in the structure of the microbial composition in the hemolymph, hepatopancreas, and intestines of the crayfish. The abundance of Aeromonas in the hemolymph of the sick crayfish was as high as 99.33%. In particular, KEGG metabolic pathway analysis showed that some antibiotic synthesis, enterobactin biosynthesis, and myo-inositol degradation pathways were abundant in healthy crayfish hemolymphs, which may be the mechanism of maintaining crayfish health. Conversely, inhibition of these pathways led to the disorder of microbiota structure, finally leading to the occurrence of diseases. To the knowledge of the authors, this study was the first to use high-throughput amplicon sequencing targeting the 16S rRNA gene to find the causative bacteria in aquatic animals. This protocol can provide more comprehensive and reliable evidence for pathogen identification, even if the pathogenic bacteria are anaerobes or other hard-to-culture bacteria.

Effects of bentonite on the growth process of submerged macrophytes and sediment microenvironment.

The effects of clay mineral bentonite on the growth process of submerged macrophyte V. spiralis and sediment microenvironment were investigated in the study for the first time, aiming to determine whether it is suitable for application in the field of ecological restoration. The growth index, and physiological and biochemical index of V. spiralis in the experiments were measured once a month, and the changes of rhizosphere microorganisms and physicochemical properties of sediments were also studied at the same time. The results demonstrated that bentonite can effectively promote the growth of V. spiralis. The treatment groups of RB1/1 and MB1/5 (the mass ratios of bentonite to sediment were 1/1 and 1/5, respectively.) showed the best V. spiralis growth promotion rates which were 18.78%, and 11.79%, respectively. The highest microbial diversity and abundance existed in group of RB10 (the mass ratio of sediment to bentonite was 10/1), in which the OTUs, Shannon, Chao and Ace were 1521.0, 5.20, 1712.26, and 1686.31, respectively. Bentonite was conducive to the propagation of rhizosphere microorganisms, and further changed the physical and chemical properties of the sediment microenvironment. The nutrient elements dissolved from bentonite may be one of the main reasons that promoted the growth of V. spiralis. The purpose of this result is to prove that bentonite can be further applied as sediment improver and growing media in ecological restoration projects in eutrophic shallow lakes.

Effects of plant on denitrification pathways in integrated vertical-flow constructed wetland treating swine wastewater.

Plant is an important part of constructed wetland (CW), while, its potential effect on nitrogen cycling is complicated. Herein, integrated vertical-flow constructed wetland (IVCW) in pilot-scale planted with Arundo donax (Planted System, PS) was constructed to treat swine wastewater. The removal performance of nitrogen in PS, effects of plant on the microbial community structure and nitrogen related function genes were revealed. Results showed that, Arundo donax planting enhanced the removal rate of TN, compared to unplanted IVCWs, the absolute abundance of Pseudomonas, Acinetobacter and Bacillus in PS was significantly increased, as well as the absolute abundance of functional gene (amoA, nxrA, nirK, nirS and nosZ). The denitrification process was mainly occurred in down-flow cell of PS with significantly higher abundant of nirK and nosZ (P < 0.05). These findings suggested that Arundo donax planting in IVCWs with zeolite as substrate promoted the growth of denitrifying microorganisms under higher pollutant load. In addition, the increased abundant of nosZ and the ratio of nosZ/∑nir indicating a lower genetic potential for N2O release. Our research provides new insight into the potential application of plant on the purification of swine wastewater.

Effects of maifanite on growth, physiological and phytochemical process of submerged macrophytes Vallisneria spiralis.

The restoration of submerged plants is critical for the reconstruction of eutrophic lake ecosystems. The growth of submerged plants is influenced by many factors. For the first time in this study, the effects of silicate-mineral maifanite supplement on the growth, physiological and phytochemical process of Vallisneria spiralis (V. spiralis) were investigated by an outdoor PVC barrel experiment, to provide a technical reference for further applications in aquatic ecological restoration. The results show that the maifanite could significantly promote the growth of V. spiralis. Specifically, the biomass, height, number of leaves, leaf width, root length, and root activity of V. spiralis in the maifanite-supplemented group were better than those of the control (P < 0.05). Moreover, the modified maifanite group performed better than the raw maifanite group (P < 0.05). The photosynthetic pigment, root activity, and the malondialdehyde and peroxidase activity of the maifanite-treated V. spiralis were better than those of the control to some extent. It was found that maifanite contained abundant major and trace elements, which are required for the growth of V. spiralis. It is concluded that maifanite is beneficial to the growth of V. spiralis and can be further applied to the ecological restoration of eutrophic lakes.

Enhanced nitrate reduction in water by a combined bio-electrochemical system of microbial fuel cells and submerged aquatic plant Ceratophyllum demersum.

High nitrate (NO3-) loading in water bodies is a crucial factor inducing the eutrophication of lakes. We tried to enhance NO3- reduction in overlying water by coupling sediment microbial fuel cells (SMFCs) with submerged aquatic plant Ceratophyllum demersum. A comparative study was conducted by setting four treatments: open-circuit SMFC (Control), closed-circuit SMFC (SMFC-c), open-circuit SMFC with C. demersum (Plant), and closed-circuit SMFC with C. demersum (P-SMFC-c). The electrochemical parameters were documented to illustrate the bio-electrochemical characteristics of SMFC-c and P-SMFC-c. Removal pathways of NO3- in different treatments were studied by adding quantitative 15NO3- to water column. The results showed that the cathodic reaction in SMFC-c was mainly catalyzed by aerobic organisms attached on the cathode, including algae, Pseudomonas, Bacillus, and Albidiferax. The oxygen secreted by plants significantly improved the power generation of SMFC-c. Both electrogenesis and plants enhanced the complete removal of NO3- from the sediment-water system. The complete removal rates of added 15N increased by 17.6% and 10.2% for SMFC-c and plant, respectively, when compared with control at the end of experiment. The electrochemical/heterotrophic and aerobic denitrification on cathodes mainly drove the higher reduction of NO3- in SMFC-c and plant, respectively. The coexistence of electrogenesis and plants further increased the complete removal of NO3- with a rate of 23.1%. The heterotrophic and aerobic denitrifications were simultaneously promoted with a highest abundance of Flavobacterium, Bacillus, Geobacter, Pseudomonas, Rhodobacter, and Arenimonas on the cathode.

Both widespread PEP-CTERM proteins and exopolysaccharides are required for floc formation of Zoogloea resiniphila and other activated sludge bacteria.

Bacterial floc formation plays a central role in the activated sludge (AS) process, which has been widely utilized for sewage and wastewater treatment. The formation of AS flocs has long been known to require exopolysaccharide biosynthesis. This study demonstrates an additional requirement for a PEP-CTERM protein in Zoogloea resiniphila, a dominant AS bacterium harboring a large exopolysaccharide biosynthesis gene cluster. Two members of a wide-spread family of high copy number-per-genome PEP-CTERM genes, transcriptionally regulated by the RpoN sigma factor and accessory PrsK-PrsR two-component system and at least one of these, pepA, must be expressed for Zoogloea to build the floc structures that allow gravitational sludge settling and recycling. Without PrsK or PrsR, Zoogloea cells were planktonic rather than flocculated and secreted exopolysaccharides were released into the growth broth in soluble form. Overexpression of PepA could circumvent the requirement of rpoN, prsK and prsR for the floc-forming phenotype by fixing the exopolysaccharides to bacterial cells. However, overexpression of PepA, which underwent post-translational modifications, could not rescue the long-rod morphology of the rpoN mutant. Consistently, PEP-CTERM genes and exopolysaccharide biosynthesis gene cluster are present in the genome of the floc-forming Nitrospira comammox and Mitsuaria strain as well as many other AS bacteria.

Bacterial Communities and Their Predicted Functions Explain the Sediment Nitrogen Changes Along with Submerged Macrophyte Restoration.

Submerged vegetation biomass fluctuation usually occurs during the preliminary stage of vegetation restoration in shallow lakes, which impacts the final status and duration for achieving a macrophyte-dominant state. This study uncovered the sediment N characteristics and the sediment bacterial community and their predicted functions during the preliminary stage of vegetation recovery in the West Lake, a typical subtropical degenerated shallow lake in China. Results showed increased amounts of sediment TN and NH4-N, reaching 3425.76 and 345.5 mg kg-1, respectively, when the vegetation biomass decreased from its maximum to its minimum. The maximum concentration of sediment NH4-N reached 508.60 mg kg-1 with the decline in vegetation, which might restrict further growth of the submerged macrophytes. The bacterial community structure during the high macrophyte biomass (HMB) period was distinct from that observed during the low macrophyte biomass (LMB) period. Specific taxa such as the phyla Chloroflexi and Acidobacteria and the genus Anaerolineaceae that are related to organic carbon degradation were significantly higher during the LMB period. Potential denitrifiers, such as Lactococcus and Bacillus genera decreased during the LMB period. Accumulation of sediment ammonia could be attributed to the enhanced production by assimilatory nitrate reduction, organic N degradation, and/or the decreased consumption by nitrification. Our findings highlight that the unstable preliminary stage of vegetation restoration brings drastic fluctuation of sediment N loading, of which NH4-N accumulation caused by bacterial communities prevents further growth of the submerged macrophytes. Therefore, extra management measures for the vegetation recovery areas should be taken to avoid excess NH4-N accumulation in sediments.

Effects of the decomposing liquid of Cladophora oligoclona on Hydrilla verticillata turion germination and seedling growth.

Excessive proliferation of filamentous green algae (FGA) has been considered an important factor resulting in the poor growth or even decline of submerged macrophytes. However, there is a lack of detailed information regarding the effect of decaying FGA on submerged macrophytes. This study aimed to investigate whether the decomposing liquid from Cladophora oligoclona negatively affects Hydrilla verticillata turion germination and seedling growth. The results showed that the highest concentrations of decomposing liquid treatments inhibited the turion germination rate, which was the lowest than other treatments, at only 84%. The chlorophyll a fluorescence (JIP test) and physiological indicators (chlorophyll a content, soluble sugars, Ca2+/Mg2+-ATPase and PAL activity) were also measured. The chlorophyll a content in the highest concentration (40% of original decomposing liquid) treatment group decreased by 43.53% than that of the control; however, soluble sugars, Ca2+/Mg2+-ATPase, and PAL activity increased by 172.46%, 271.19%, and 26.43% respectively. The overall results indicated that FGA decay has a considerable effect on submerged macrophyte turion germination and seedling growth, which could inhibit their expansion and reproduction. This study emphasized the need to focus on effects of FGA decomposition on the early growth stages of submerged macrophytes and offered technological guidance for submerged vegetation restoration in lakes and shallow waters.

Relationship between electrogenic performance and physiological change of four wetland plants in constructed wetland-microbial fuel cells during non-growing seasons.

To find suitable wetland plants for constructed wetland-microbial fuel cells (CW-MFCs), four commonly used wetland plants, including Canna indica, Cyperus alternifolius L., Acorus calamus, and Arundo donax, were investigated for their electrogenic performance and physiological changes during non-growing seasons. The maximum power output of 12.82mW/m2 was achieved in the A. donax CW-MFC only when root exudates were being released. The results also showed that use of an additional carbon source could remarkably improve the performance of electricity generation in the C. indica and A. donax CW-MFCs at relatively low temperatures (2-15°C). However, A. calamus withered before the end of the experiment, whereas the other three plants survived the winter safely, although their relative growth rate values and the maximum quantum yield of PSII (Fv/Fm) significantly declined, and free proline and malondialdehyde significantly accumulated in their leaves. On the basis of correlation analysis, temperature had a greater effect on plant physiology than voltage. The results offer a valuable reference for plant selection for CW-MFCs.

RpoN (σ54) Is Required for Floc Formation but Not for Extracellular Polysaccharide Biosynthesis in a Floc-Forming Aquincola tertiaricarbonis Strain.

Some bacteria are capable of forming flocs, in which bacterial cells become self-flocculated by secreted extracellular polysaccharides and other biopolymers. The floc-forming bacteria play a central role in activated sludge, which has been widely utilized for the treatment of municipal sewage and industrial wastewater. Here, we use a floc-forming bacterium, Aquincolatertiaricarbonis RN12, as a model to explore the biosynthesis of extracellular polysaccharides and the regulation of floc formation. A large gene cluster for exopolysaccharide biosynthesis and a gene encoding the alternative sigma factor RpoN1, one of the four paralogues, have been identified in floc formation-deficient mutants generated by transposon mutagenesis, and the gene functions have been further confirmed by genetic complementation analyses. Interestingly, the biosynthesis of exopolysaccharides remained in the rpoN1-disrupted flocculation-defective mutants, but most of the exopolysaccharides were secreted and released rather than bound to the cells. Furthermore, the expression of exopolysaccharide biosynthesis genes seemed not to be regulated by RpoN1. Taken together, our results indicate that RpoN1 may play a role in regulating the expression of a certain gene(s) involved in the self-flocculation of bacterial cells but not in the biosynthesis and secretion of exopolysaccharides required for floc formation.IMPORTANCE Floc formation confers bacterial resistance to predation of protozoa and plays a central role in the widely used activated sludge process. In this study, we not only identified a large gene cluster for biosynthesis of extracellular polysaccharides but also identified four rpoN paralogues, one of which (rpoN1) is required for floc formation in A. tertiaricarbonis RN12. In addition, this RpoN sigma factor regulates the transcription of genes involved in biofilm formation and swarming motility, as previously shown in other bacteria. However, this RpoN paralogue is not required for the biosynthesis of exopolysaccharides, which are released and dissolved into culture broth by the rpoN1 mutant rather than remaining tightly bound to cells, as observed during the flocculation of the wild-type strain. These results indicate that floc formation is a regulated complex process, and other yet-to-be identified RpoN1-dependent factors are involved in self-flocculation of bacterial cells via exopolysaccharides and/or other biopolymers.

Effects of Planted Versus Naturally Growing Vallisneria natans on the Sediment Microbial Community in West Lake, China.

Submerged macrophytes play an important role in aquatic ecosystems, which has led to an increase in studies on vegetation recovery in polluted lakes from which submerged macrophytes have disappeared. The comparison of microbial communities in sediment cloned with planted and naturally growing submerged macrophytes is an interesting but rarely studied topic. In this investigation, Maojiabu and Xilihu, two adjacent sublakes of West Lake (Hangzhou, China), were selected as aquatic areas with planted and naturally growing macrophytes, respectively. Sediment samples from sites with/without Vallisneria natans were collected from both sublakes. The results showed that sediment total nitrogen and organic matter were significantly lower in the plant-covered sites than that in the non-plant sites in Maojiabu. Additionally, the sediment microbial community characterized by 16S ribosomal RNA (rRNA) sequencing differed more significantly for Maojiabu than for Xilihu. The relative abundances of microbes involved in C, N, and S elemental cycling were significantly higher in the sediments with plants than in those without. Results from both fatty acid methyl ester analysis and 16S rRNA sequencing indicated that vegetation significantly influenced the sulfate-reducing bacteria (SRB). Thus, the gene copies and composition of SRB were explored further. The relative gene abundance of SRB was 66% higher with natural vegetation colonization but was not influenced by artificial colonization. An increase in dominant SRB members from the families Syntrophobacteraceae and Thermodesulfovibrionaceae contributed to the increase of total SRB. Thus, macrophyte planting influences sediment nutrient levels and microbial community more than natural growth does, whereas the latter is more beneficial to sediment SRB.

Impacts of residual aluminum from aluminate flocculant on the morphological and physiological characteristics of Vallisneria natans and Hydrilla verticillata.

Aluminate is generally used as a flocculant in water and wastewater treatment processes, but the residual aluminum (Al) may have toxic effects on aquatic organisms when the concentration accumulates beyond a threshold level. The in situ and laboratory tests were conducted to evaluate the impact of residual Al on submerged macrophytes in West Lake, Hangzhou, China, which receives Al flocculant-purified water diverted from the Qiantang River. The responses of Vallisneria natans and Hydrilla verticillata were investigated based on their morphological and physiological parameters in pot culture and aquarium simulation experiments. In the pot culture experiments, the biomass, seedling number, plant height, stolon number, stolon length, and root weight were significantly higher at a site located 150m from the inlet compared with those at a site located 15m from the inlet (P < 0.05), thereby indicating that the residual Al significantly inhibited the morphological development of V. natans and H. verticillata. The variations in the chlorophyll-a, protein, and malondialdehyde contents of the two species in both the pot culture and aquarium simulation experiments also demonstrated that the two submerged macrophytes were stressed by residual Al. V. natans and H. verticillata accumulated 0.052-0.227mg of Al per gram of plant biomass (fresh weight, mg/g FW) and 0.045-0.205mg Al/g FW in the in situ experiments, respectively, where the amounts of Al were significantly higher in the plants in the treatment aquaria during the laboratory experiments than those in the controls. These results may have important implications for the restoration of submerged macrophytes and ecological risk assessments in Al-exposed lakes. It is recommended that the Al salt concentration used for the control of lake eutrophication should be reduced to an appropriate level.

Effects of pyrogallic acid on Microcystis aeruginosa: oxidative stress related toxicity.

Pyrogallic acid (PA) is used in various industrial and consumer products. The molecular mechanisms underlying PA's toxicity was not fully understood. In this study, toxicity of PA on Microcystis aeruginosa with reactive oxygen species (ROS) generation as an end point was investigated. The results showed an increase in the percentage of cells with loss of membrane integrity and enhanced intracellular ROS production. Exposure to 50mgL(-1) PA for 48h caused the highest percentage of loss of membrane integrity (56.7%), and a 2.54-fold higher intracellular ROS level compared to control. Further investigation revealed that PA caused a dose-dependent increase in DNA strand breaks (DSB) of M. aeruginosa at exposure concentration from 2 to 50mgL(-1). The incubation of cells with ROS scavengers ascorbic acid, N-acetyl-l-cysteine (NAC) and tocopherol markedly alleviated the level of PA-induced DSB. Analysis of PA autoxidized products in culture solution showed that PA was quickly converted to purpurogallin (PG), and PG was further autoxidized to other polyphenolic compounds. PA and PG might participate a futile redox cycle, which mediated ROS production in M. aeruginosa. These results suggested DNA strands and cell membrane were two targets of ROS induced by PA, and oxidative damage was an important mechanism for the toxicity of PA against M. aeruginosa.