Azoarcus halotolerans sp. nov., a novel member of Rhodocyclaceae isolated from activated sludge collected in Hong Kong.

A floc-forming bacterial strain, designated HKLI-1T, was isolated from the activated sludge of a municipal sewage treatment plant in Hong Kong SAR, PR China. Cells of this strain were Gram-stain-negative, strictly aerobic, catalase- and oxidase-positive, rod-shaped and motile by means of a single polar flagellum. Growth occurred at 18-37 °C (optimum, 28 °C), pH 5.5-9.0 (optimum, pH 7.5) and with 0-8.0 % (w/v) NaCl (optimum, 1-1.5 %) concentration. The major fatty acids of strain HKLI-1T were C16 : 0 and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c). The polar lipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and three unidentified lipids. The DNA G+C content was 63.5 mol% from whole genomic sequence analysis. Based on the results of 16S rRNA gene sequences analysis, this strain should be assigned to the genus Azoarcus and is closely related to Azoarcus olearius DQS-4T (94.93 % 16S rRNA gene sequence pairwise similarity), Azoarcus toluclasticus MF63T (94.91 %) and Azoarcus communis SWub3T (94.01 %), but separate from them by large distances in different phylogenetic trees. Based on whole genome analysis, the orthologous average nucleotide identity and in silico DNA-DNA hybridization values against four of the closest relatives were 73.03-74.83 and 17.2-23.0 %, respectively. The phylogenetic, genotypic, phenotypic and chemotaxonomic data demonstrated that strain HKLI-1T could be distinguished from its phylogenetically related species, and that this strain represented a novel species within the genus Azoarcus, for which the name Azoarcus halotolerans sp. nov. is proposed. The type strain is HKLI-1T (= 72659T=CCTCC AB 2019312T).

The structure of denitrifying microbial communities in constructed mangrove wetlands in response to fluctuating salinities.

In this study, the experimental vertical-flow constructed wetland (CW) systems planted with the salt-tolerant mangrove species Kandelia candel were established to investigate the influence of salinity fluctuations on the denitrification performance and denitrifying microbial community structure of the CWs. The high-throughput sequencing analysis showed that 10-13 genera aerobic microbes had been enriched in the upper layer of wetland matrix in the depth of 10-25 cm, with the relative abundance accounting for 19.1 ±â€¯7.9%. Although the ammonium oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) were inhibited significantly in the CW systems with salinity levels in the range of 0.9-1.8%, the aerobic denitrifying (AD) bacteria including Pseudomonas, Acinetobacter and Aeromonas, removed 99% of ammonia nitrogen from the influent by heterotrophic nitrification (HN) functions, and conducted denitrification at the same time to remove 90% of the TN in the system, indicating that the wetland test system successfully enriched a variety of aerobic denitrifying bacterial communities under different salinity conditions. Not only the nitrogen removal efficiency but also the adaptability of the wetland system to salinity fluctuations had been improved by the enriched HN-AD bacteria. In addition, HN-AD bacterial communities can conduct both nitrification and denitrification in the middle and upper layers of the vertical flow wetland, hereby saving the reaction space of the constructed wetland and reducing the construction cost.

A vertical-flow constructed wetland-microalgal membrane photobioreactor integrated system for treating high-pollution-load marine aquaculture wastewater: A lab-scale study.

Individual biological water treatment techniques often prove ineffective in removing accumulated high concentrations of nitrogen and phosphorus in the late stages of biofloc aquaculture. To address this issue, we integrated a previously developed autotrophic denitrification and nitrification integrated constructed wetland (ADNI-CW) with a microalgal membrane photobioreactor (MPBR). Under high nitrogen and phosphorus pollution loads in the influent, the standalone ADNI-CW system achieved removal rates of only 24.17 % ± 2.82 % for total nitrogen (TN) and 25.30 % ± 2.59 % for total phosphorus (TP). The optimal conditions for TN and TP degradation and microalgal biomass production in the Chlorella MPBR, determined using response surface methodology, were an inoculum OD680 of 0.394, light intensity of 161.583 µmol/m2/s, and photoperiod of 16.302 h light:7.698 h dark. Under the optimal operating conditions, the integrated ADNI-CW-MPBR system achieved remarkable TN and TP removal rates of 92.63 % ± 2.8 % and 77.46 % ± 8.41 %, respectively, and a substantial microalgal biomass yield of 54.58 ± 6.8 mg/L/day. This accomplishment signifies the successful achievement of efficient nitrogen and phosphorus removal from high-pollution-load marine aquaculture wastewater along with the acquisition of valuable microalgal biomass. A preliminary investigation of the microbial community composition and algal-bacterial interactions in different operational stages of the MPBR system revealed that unclassified_d__Bacteria, Chlorophyta, and Planctomycetes were predominant phyla. The collaborative relationships between bacteria and Chlorella surpassed competition, ensuring highly efficient nitrogen and phosphorus removal in the MPBR system. This study laid the foundation for the green and sustainable development of the aquaculture industry.

The Transcriptome of Brassica napus L. Roots under Waterlogging at the Seedling Stage.

Although rapeseed (Brassica napus L.) is known to be affected by waterlogging, the genetic basis of waterlogging tolerance by rapeseed is largely unknown. In this study, the transcriptome under 0 h and 12 h of waterlogging was assayed in the roots of ZS9, a tolerant variety, using digital gene expression (DGE). A total of 4432 differentially expressed genes were identified, indicating that the response to waterlogging in rapeseed is complicated. The assignments of the annotated genes based on GO (Gene Ontology) revealed there were more genes induced under waterlogging in "oxidation reduction", "secondary metabolism", "transcription regulation", and "translation regulation"; suggesting these four pathways are enhanced under waterlogging. Analysis of the 200 most highly expressed genes illustrated that 144 under normal conditions were down-regulated by waterlogging, while up to 191 under waterlogging were those induced in response to stress. The expression of genes involved under waterlogging is mediated by multiple levels of transcriptional, post-transcriptional, translational and post-translational regulation, including phosphorylation and protein degradation; in particular, protein degradation might be involved in the negative regulation in response to this stress. Our results provide new insight into the response to waterlogging and will help to identify important candidate genes.

A novel autotrophic denitrification and nitrification integrated constructed wetland process for marine aquaculture wastewater treatment.

We undertook a lab-scale evaluation of a novel autotrophic denitrification and nitrification integrated constructed wetland (ADNI-CW) for improved carbon (C), nitrogen (N), and sulfur (S) cycling to treat mariculture wastewater. The process involved an up-flow autotrophic denitrification constructed wetland unit (AD-CW) for sulfate reduction and autotrophic denitrification, and an autotrophic nitrification constructed wetland unit (AN-CW) for nitrification. The 400-day experiment investigated the performance of the AD-CW, AN-CW, and entire ADNI-CW processes under various hydraulic retention times (HRTs), nitrate concentrations, dissolved oxygen levels, and recirculation ratios. Under various HRTs, the AN-CW achieved a nitrification performance exceeding 92%. Correlation analysis of the chemical oxygen demand (COD) revealed that, on average, approximately 96% of COD was removed by sulfate reduction. Under different HRTs, increases in influent NO3--N concentrations caused the amount of sulfide to gradually decrease from sufficient to deficient, and the autotrophic denitrification rate also decreased from 62.18 to 40.93%. In addition, when the NO3--N load rate was above 21.53 g N/m2·d, the transformation of organic N by mangrove roots may have increased NO3--N in the top effluent of the AD-CW. The coupling of N and S metabolic processes mediated by various functional microorganisms (Proteobacteria, Chloroflexi, Actinobacteria, Bacteroidetes, and unclassified_d__Bacteria) enhanced N removal. We intensively explored the effects of changing inputs as culture species developed on the physical, chemical, and microbial changes of CW to ensure a consistent and effective management of C, N, and S. This study lays the foundation for green and sustainable mariculture development.

Effects of different aeration strategies and ammonia-nitrogen loads on nitrification performance and microbial community succession of mangrove constructed wetlands for saline wastewater treatment.

In highly salinized environments, nitrification is the process that limits the rate of nitrogen transformation and removal. Therefore, this study concentrated on the impacts of different aeration strategies and NH4+-N loads on the nitrification performance of mangrove constructed wetlands (CWs), as well as investigating the succession mechanism of ammonia-oxidizing microorganisms (AOMs). The results showed that both the CW with continuous aeration (CA-CW) and intermittent aeration (IA-CW) achieved a nitrification efficiency of more than 98% under an NH4+-N loading of 1.25-4.7 g/(m2·d). However, the total nitrogen removal rates of IA-CW under low and high ammonia-nitrogen loads (LAL, 20.09 ± 4.4% and HAL, 8.77 ± 1.35%, respectively) were higher than those of CA-CW (16.11 ± 4.7% and 3.32 ± 2.3%, respectively), especially under HAL (p < 0.05). Pearson correlation analysis showed that under different operating conditions, the differential secretion of Kandelia candel rhizosphere organic matter had a certain regulatory effect on nitrification and denitrification groups such as Candidatus Nitrocosmicus, Nitrancea, Truepera, Pontibacter, Halomonas, and Sulfurovum in the wetland root layer. The quantitative polymerase chain reaction revealed that the NH4+-N load rate was the primary factor driving the succession of the AOMs, with different aeration strategies exacerbating this process. Overall, this study revealed that the dominant AOMs in mangrove CWs could be significantly altered by regulating the aeration modes and pollution loads to adjust the rhizosphere organic matter in situ, thereby resulting in more efficient nitrification.

Biochar immobilized bacteria enhances nitrogen removal capability of tidal flow constructed wetlands.

To improve the nitrogen removal (NR) capability of tidal flow constructed wetlands (TFCWs) for treatment of saline wastewater, biochar, produced from Cyperus alternifolius, was used to adsorb and immobilize a salt tolerant aerobic denitrifying bacteria (Zobellella sp. A63), and then was added as a substrate into the systems. Under low (2:1) or high (6:1) C/N ratio, the removal of NO3--N and total nitrogen (TN) in the biochar immobilized bacteria (BIB) dosing system (TFCW3) was significantly higher (q < 0.05) than that in the untreated system (TFCW1) and the biochar dosing system (TFCW2). At low C/N ratio, the removal rates of NO3--N, TN and chemical oxygen demand (COD) of TFCW3 were 68.2%, 72.6% and 82.5%, respectively, 15-20% higher than TFCW1 and 5-10% higher than TFCW2. When C/N ratio was further increased to 6, the pollutant removal rate of each system was greatly improved, but the removal rate of TFCW3 for NO3--N/TN was still nearly 10% and 5% higher than TFCW1 and TFCW2, respectively. Microbial community analysis showed that aerobic denitrifying bacteria, sulfate reducing bacteria and sulfur-driven denitrifiers (DNSOB) played the most important role of NR in TFCWs. Moreover, biochar bacterial agent significantly increased the abundances of genes involved in NR. The total copy numbers of bacterial 16S rRNA, nirS, nirK, drsA and drsB genes in the TFCW3 were 1.1- to 3.76-fold higher than those in the TFCW1; Especially at low C/N ratio, the copy number of drsA and drsB in the upper layer of TFCW3 were 85.5 and 455 times that of TFCW1, respectively. Thus, BIB provide a more feasible and effective amendment for constructed wetlands to improve the N removal of the saline wastewater by enhancing the microbial NR capacity mainly via aerobic and sulfur autotrophic denitrification.

Portulaca oleracea polysaccharides reduce serum lipid levels in aging rats by modulating intestinal microbiota and metabolites.

Metabolic diseases characterized by dyslipidemia are common health problems for elderly populations. Dietary fiber intake is inversely associated with the risk of dyslipidemia. This study investigated the effects of Portulaca oleracea polysaccharide (POP) on the intestinal microbiota and its metabolites in aging rats using 16S rRNA sequencing and metabolomics techniques. Our results showed that POPs reduced the ratio of Firmicutes/Bacteroidetes (F/B), relative abundance of Fusobacteria, and levels of triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), and gamma-glutamyl transferase (γ-GT) in the serum of aging rats. POP supplementation also reduced 5beta-cholestane-3alpha,7alpha,12alpha,25-tetrol, and vaccenic acid concentrations in lipids and lipoid-like molecules, while soyasapogenol E and monoacylglycerol (MG) (24:0/0:0/0:0) levels increased. This study demonstrated that POP's beneficial effects on lipid levels in aging rats might be partially attributable to the modification of gut microbiota and related metabolites.

Bioaugmented constructed wetlands for efficient saline wastewater treatment with multiple denitrification pathways.

Six laboratory-scale constructed wetlands (CWs) were used to quantify the nitrogen removal (NR) capacity in the treatment of saline wastewater at high (6:1) and low (2:1) carbon-nitrogen ratios (C/N), with and without bioaugmentation of aerobic-denitrifying bacterium. Sustained high-efficiency nitrification was observed throughout the operation. However, under different C/N ratios, although the bioaugmentation of aerobic-denitrifying bacterium promoted the removal of NO3--N and TN, there were still great differences in denitrification. Molecular biology experiments revealed ammonia-oxidizing archaea, together with the Nitrosomonas and Nitrospira, led to highly efficient nitrification. Furthermore, aerobic-denitrifying bacterium and sulfur-driven denitrifiers were the core denitrification groups in CWs. By performing these combined experiments, it was possible to determine the optimal CW design and the most relevant NR processes for the treatment of salty wastewater. The results suggest that the bioaugmentation of salt-tolerant functional bacteria with multiple NR pathways are crucial for the removal of salty wastewater pollutants.

Effects of substrate type on denitrification efficiency and microbial community structure in constructed wetlands.

Three constructed wetland systems were established to treat saline sewage via high-porosity ceramsite, activated carbon, and low-porosity sand: A (ceramsite + activated carbon + sand), B (sand + activated carbon + ceramsite), and C (sand). The distribution of dissolved oxygen in these systems varied with different filling methods with the best removal efficiency of ammonium nitrogen and total nitrogen observed in system B (97.4 and 96.2%, respectively). The 16S rDNA amplicon sequencing results showed that all the systems had a high abundance of salt-tolerant denitrifiers, and the filling method significantly impacted denitrifying bacteria (e.g., Vibrio and Planctomyces) in the substrate. System B had more diverse dissolved oxygen conditions than system A and showcased aerobic nitrification-denitrification and anaerobic ammonium oxidation pathways. Therefore, the use of substrates with different porosities can improve the dissolved oxygen supply and enhance nitrogen removal efficiency in constructed wetlands.

Isolation and identification of a salt-tolerant aerobic denitrifying bacterial strain and its application to saline wastewater treatment in constructed wetlands.

A salt-tolerant aerobic denitrifying bacterium, Zobellella denitrificans strain A63, was isolated, and its effects on the efficiency of denitrification of saline wastewater and the denitrifying microbial community structure in the matrix were studied in vertical-flow constructed wetlands (VFCWs). In a VFCW system with strain A63, the removal efficiencies of NH4+-N, NO3--N, and total nitrogen reached 79.2%, 95.7%, and 89.9%, respectively. Quantitative PCR analysis indicated that the amoA gene from ammonia-oxidizing archaea (AOA) was highly abundant, whereas amoA from ammonia-oxidizing bacteria and nxrA from nitrite-oxidizing bacteria were lowly abundant because of the influent salinity, irrespective of whether strain A63 was added. However, the addition of strain A63 significantly increased the abundance of nirK in the top layer of the VFCW. Therefore, AOA-driven partial nitrification and aerobic denitrification by strain A63 occurred in VFCWs. Our findings suggest that adding salt-tolerant denitrifying strains to constructed wetlands can enhance denitrification for saline wastewater treatment.

Effects of different potassium fertilizers on cadmium uptake by three crops.

Cadmium contamination of agricultural soils has aroused worldwide concern because of the threats posed to human health through accumulation in food chains. A greenhouse pot experiment was conducted with in situ Cd-contaminated soil to study the influence of different potassium fertilizers (KCl, K2SO4, and KNO3) on Cd accumulation in rice, wheat, and pak choi as well as the NH4NO3-extractable Cd (NEX-Cd) content in soils. In our study, rice and wheat biomass increased in the presence of K fertilizers, whereas pak choi biomass remained stable. Moreover, our experiment demonstrated that Cl- increased Cd uptake by crops more effectively than SO42- or NO3-. The KCl treatments increased the Cd content of all three crops; as the KCl dose was increased, the Cd content of rice grains, wheat grains, and pak choi shoots increased by 10.8-192.8%, 17.1-67.7%, and 15.1-40.4%, respectively. The KNO3 treatment also increased the Cd content of all three crops; however, the K2SO4 treatment only slightly increased the Cd content of wheat and pak choi and greatly decreased the Cd content of rice. In addition, both of the NEX-Cd content of wheat soil and pak choi soil were much higher than that of rice paddy soil. The KCl treatment resulted in a significant increase in the NEX-Cd content of rice paddy soil, but there were no significant differences in the NEX-Cd content of wheat soil or pak choi soil, regardless of which types or doses of K fertilizers were supplied. Based on these results, when K fertilizers are applied to Cd-contaminated soils, both types and doses should be carefully considered to mitigate Cd accumulation in crops, especially the edible part.

The influence of complex fermentation broth on denitrification of saline sewage in constructed wetlands by heterotrophic nitrifying/aerobic denitrifying bacterial communities.

An experimental vertical-flow constructed wetland (CW) was tested to treat salt-containing sewage. CW clogging deposits and withered Pontederia cordata L. were collected into a complex fermentation broth to serve as the carbon source and its effects on the denitrification capacity and microbial composition of the CW were examined. Addition of the complex fermentation broth into the CW influent (1.8% salinity) led to high removal efficiencies of NH4+-N > 99.82 ±â€¯0.00% and TN > 90.39 ±â€¯0.05%. Heterotrophic nitrifiers and aerobic denitrifiers were entirely dominant in the middle and upper layers of the CW, where obligate halophilic, aerobic denitrifiers Zobellella occurred. The CW successfully cultivated and enriched heterotrophic nitrifying-aerobic denitrifying bacteria, overcoming the effects of salinity and insufficient organic carbon sources on the denitrification capacity of CW. This type of complex carbon sources can also facilitate the utilization of waste resources, such as CW clogging deposits and withered wetland plants.

Proteomic Analysis of Rapeseed Root Response to Waterlogging Stress.

The overall health of a plant is constantly affected by the changing and hostile environment. Due to climate change and the farming pattern of rice (Oryza sativa) and rapeseed (Brassica napus L.), stress from waterlogging poses a serious threat to productivity assurance and the yield of rapeseed in China's Yangtze River basin. In order to improve our understanding of the complex mechanisms behind waterlogging stress and identify waterlogging-responsive proteins, we firstly conducted iTRAQ (isobaric tags for relative and absolute quantification)-based quantitative proteomic analysis of rapeseed roots under waterlogging treatments, for both a tolerant cultivar ZS9 and sensitive cultivar GH01. A total of 7736 proteins were identified by iTRAQ, of which several hundred showed different expression levels, including 233, 365, and 326 after waterlogging stress for 4H, 8H, and 12H in ZS9, respectively, and 143, 175, and 374 after waterlogging stress for 4H, 8H, and 12H in GH01, respectively. For proteins repeatedly identified at different time points, gene ontology (GO) cluster analysis suggested that the responsive proteins of the two cultivars were both enriched in the biological process of DNA-dependent transcription and the oxidation⁻reduction process, and response to various stress and hormone stimulus, while different distribution frequencies in the two cultivars was investigated. Moreover, overlap proteins with similar or opposite tendencies of fold change between ZS9 and GH01 were observed and clustered based on the different expression ratios, suggesting the two genotype cultivars exhibited diversiform molecular mechanisms or regulation pathways in their waterlogging stress response. The following qRT-PCR (quantitative real-time polymerase chain reaction) results verified the candidate proteins at transcription levels, which were prepared for further research. In conclusion, proteins detected in this study might perform different functions in waterlogging responses and would provide information conducive to better understanding adaptive mechanisms under environmental stresses.

Effect of plant-based carbon sources on denitrifying microorganisms in a vertical flow constructed wetland.

The effects of supplementing plant-based carbon sources, fermented tissues of Arundo donax and Pontederia cordata, and a combination of the two plants, on the nitrogen removal efficiency and microbial composition in a vertical flow constructed wetland (VFCW) were examined. The results showed that the addition of the composite carbon source produced the highest removal efficiencies of NH4+-N 91.5%, NO3--N 94.5% and TN 92.8% in VFCW. The detected abundance of amoA, nirS, and nxrA genes indicated that ammonia oxidation bacteria and denitrifying bacteria were more abundant than the nitrite oxidation bacteria. Furthermore, the addition of the composite carbon source significantly promoted the growth of the denitrifying bacteria in VFCW. The results indicated that supplementing the system with plant-based carbon sources achieved partial nitrification and denitrification, as well as classic denitrification in VFCWs. The study suggested that multiple nitrogen removal pathways were required to feasibly and efficiently remove nitrogen.

Draft Genome Sequence of Bacillus thuringiensis NBIN-866 with High Nematocidal Activity.

Bacillus thuringiensis NBIN-866, a Gram-positive bacterium, was isolated from soil in China. We announce here the draft genome sequence of strain B. thuringiensis NBIN-866, which possesses highly nematocidal factors, such as proteins and small molecular peptides.

[Comparison of algal removal efficiencies in different ecological treatment systems and their combinations].

The algal removal efficiencies and their trends of 8 experimental ecological treatment systems, which belonged to 5 groups, were studied under different hydraulic loading rates (HLR). The algal removal efficiencies were compared according to the seasonal changes under the optimum HLR. Gravel-bed horizontal-flow wetland, down-flow wetland, up-flow wetland, aerobic ponds or facultative ponds combined these systems. The results show that the best HLR is 0.8m3/(m2 x d) in all the experimental systems. The systems including aerobic pond and down-flow wetland show a stronger resistance to hydraulic shock loading, as well as horizontal-flow wetland. The algal removal rates of all the systems peaked to over 94% in summer and bottomed in winter that varied from 34% to 90%. It was the combined form that determined the removal efficiency of the ecological treatment systems. The combination system of down-flow and up-flow wetland present the highest capacity of algal removal efficiency and lower seasonal fluctuation, while the gravel-bed horizontal-flow wetland system present the lowest efficiency and higher seasonal fluctuation.

[Temporal and spatial characteristics of substrate enzyme activities and bacteria physiological groups in constructed wetland].

Six enzymes and five bacteria physiological groups from the substrate (soil) of integrated vertical flowing constructed wetland are chosen to study temporal and spatial characteristics. The research results show that significant differences existed in each enzyme during different months (p < 0.05). The enzyme activities of cellulase, protease and phosphatase were significantly higher in June, September and December than those in March; The enzyme activity of beta-glucosidase in June was significantly higher than those in other months; While urease activities in both September and December were extremely significantly higher than those in March and in June (p < 0.01); But the dehydrogenase was different from the above-mentioned enzymes, which had higher enzyme activities in June and in December than in March and in September. Compared with the temporal characteristics, the spatial characteristics of all six enzymes show consistency: the enzyme activities in the down-flow chamber were significantly higher than those in the up-flow chamber (p < 0.05), and all of the enzyme activities in both chambers were relatively lower with the increasing depths of substrate layers. The numbers of bacteria physiological groups were at peak in June and September and their spatial characteristics were the same with enzyme activities.