Planktonic microbial community and biological metabolism in a subtropical drinking water river-reservoir system.

To understand the dynamics of planktonic microbial community and its metabolism processes in subtropical drinking water river-reservoir system with lower man-made pollution loading, this study selected Dongzhen river-reservoir system in Mulan Creek as object to investigate spatial-temporal characteristics of community profile and functional genes involved in biological metabolism, and to analyze the influence of environmental factors. The results indicated that Proteobacteria and Actinobacteria were the most diverse phyla with proportion ranges of 9%-80% in target system, and carbohydrate metabolism (5.76-7.12 × 10-2), amino acid metabolism (5.78-7.21 × 10-2) and energy metabolism (4.07-5.17 × 10-2) were found to be the dominant pathways of biological metabolism. Although there were variations in biological properties both spatially and temporally, seasonal variation had a greater influence on microbial community and biological metabolism, than locational differences. Regarding the role of environmental factors, this study revealed that microbial diversity could be affected by multiple abiotic factors, with total organic carbon, total phosphorus and temperature being more influential (absolute value of standardized regression weights >2.13). Stochastic processes dominated the microbial community assembly (R2 of neutral community model = 0.645), while niche-based processes differences represented by nutrients, temperature and pH level played secondary roles (R > 0.388, P < 0.01). Notably, the synergistic influences among the environmental factors accounted for the higher percentages of community variation (maximum proportion up to 17.6%). Additionally, pH level, temperature, and concentrations of dissolved oxygen, carbon and nitrogen were found to be the significant factors affecting carbon metabolism pathways (P < 0.05), yet only total organic carbon significantly affected on nitrogen transformation (P < 0.05). In summary, the microbial profile in reservoir is not completely dominated by that in feeding river, and planktonic microbial community and its metabolism in subtropical drinking water river-reservoir system are shaped by multiple abiotic and biotic factors with underlying interactions.

Cross-effect of wetland substrates properties on anammox process in three single-substrate anammox constructed wetlands for treating high nitrogen sewage with low C/N.

Constructing a stable and efficient anammox-driven constructed wetlands (CWs) system for efficiently treating high-nitrogen wastewater with low C/N remains a challenge, due to slow growth rate and high sensitivity of anammox bacteria to changing environmental conditions. Notably, sensitive anammox bacteria is still affected by the physicochemical properties of wetland substrates and their effects are still unknown. Therefore, three single-substrate (gravel, zeolite, and oyster shell) CWs were constructed with the goal of enhancing total nitrogen (TN) removal by anammox-driven/dominant process and determining the effect of substrate on anammox process. The gravel, zeolite and oyster shell systems achieved desired TN removal rates of 20.50, 14.25 and 22.15 g·(m2·d)-1 when influent TN load was 32.57 g·(m2·d)-1 without carbon source and costly aeration, respectively. Oyster shell system exhibited the highest removal ability and better capacity for resistance to influent nitrogen load, followed by gravel and zeolite systems (p < 0.05). Integrated analyses indicated anammox-driven/dominant process was the foremost reason accounted for the enhanced nitrogen treatment performance in all systems. The abundance of anammox gene was higher than the total abundance of denitrifying genes in the three CWs when influent TN load reached 14.85 g·(m2·d)-1. Path analysis further demonstrated anammox process was the foremost nitrogen removal pathway. [anammox] had a highest positive direct contribution (97.3%) on TN transformation rate in gravel system; [anammox/(napA+narG+nirK+nirS+nosZ)] showed highest positive direct contribution (92.4% and 97.4%) on that in zeolite and oyster shell systems, respectively. Substrate configurations significantly affected nitrogen transformation pathway and microbial communities, particularly those of anammox bacteria. Anammox genera of Candidatus Brocadia (primary anammox genera) and Candidatus Kuenenia exhibited different evolutions among the three CWs. Machine learning of Least absolute shrinkage and selection operator (LASSO) analyses showed pH, Ca, Mg, EC, and K were the key physicochemical properties of wetland substrates affecting anammox gene and anammox genera. In conclusion, Oyster shell was the optimal substrate for anammox bacteria growth.

Performance of tidal and non-tidal mangrove constructed wetlands in treating maricultural wastewater.

This study aims to evaluate the removal efficiency of nitrogen and phosphorus in the tidal and non-tidal constructed wetlands with typical mangrove (Aegiceras corniculatum) as a wetland plant model to treat simulated marine wastewater. The results showed that the average removals of NO2--N, NO3--N, NH4+-N, TN and TP were 88.4, 80.5, 81.4, 79.7 and 40.8%, respectively, in the non-tidal subsurface flow (HF) mangrove wetland, and 65.3, 61.3, 90.6, 60.1 and 19.2% in the tidal (TF) mangrove wetland, and 11.4, 64.6, 68.7, 56.6 and 16.3% in the non-tidal free water surface (FWS) mangrove wetland, respectively. Moreover, it was observed that the composition of microbial communities in the HF mangrove wetland was beneficial to the nitrogen cycle and has more quantitative associations of N-metabolism genes. The results indicated that non-tidal HF mangrove wetland has a stable and an effective capacity for potential treatment of marine wastewater compared with the non-tidal FWS mangrove wetland and tidal TF mangrove wetland.

Higher Temperatures Do Not Always Achieve Better Antibiotic Resistance Gene Removal in Anaerobic Digestion of Swine Manure.

This study employed high-throughput quantitative PCR and 16S rRNA sequencing to evaluate the effect of temperature and residual antibiotics on the dynamics of antibiotic resistance genes (ARGs) and microbial communities during anaerobic digestion of swine manure. The abundances of total ARGs and 16S rRNA genes significantly decreased in all of four treatments (25°C, 37°C, and 37°C with 50 mg of wet weight antibiotics of body weight, and 55°C). The abundances of most ARG types were significantly correlated with those of the 16S rRNA gene and transposase gene (P < 0.01). However, the abundances of total ARGs at 55°C were much higher than those of other treatments. Meanwhile, the microbial communities at 55°C, where the Streptococcus pathogen remained at a relatively high abundance and cellulose degraders and hydrogen producers, such as Ethanoligenens and Coprococcus bacteria, increased, were markedly different from those of other treatments. Redundancy analysis indicates that temperature, pH, and the genus Streptococcus had the highest explanation for ARG variation among experimental factors, chemical properties, and representative genera, respectively. Network analysis further showed that the genus Streptococcus contributed greatly to the higher ARG abundance at 55°C. The moderate antibiotic residue only caused a slight and transitory inhibition for microbially diverse populations and promotion for ARG abundance, probably due to the degradation of antibiotics and microbial adaptability. Our results clarify the cooperativity of gene transfer-related items on ARG variation and intensively prove that higher temperature cannot always achieve better ARG removal in anaerobic digestion unless pathogens and gene transfer elements are more efficiently inhibited.IMPORTANCE Antibiotic resistance genes (ARGs) are frequently detected with high abundance in manure-applied soils. Anaerobic digestion is one of widely used processes for animal waste treatment. Thus, it is critical to understand the potential of anaerobic digestion to attenuate ARGs. Although some previous studies recommended thermophilic digestion for ARG removal, they did not get sufficient evidence to support this view. The antibiotics applied to animals are mostly excreted through feces and urine because of incomplete metabolism. It is indispensable to know whether residual antibiotics in manure will hinder ARG attenuation in anaerobic digesters. The significance of our research is in comprehensively understanding the evolution and mechanism of ARGs in anaerobic digestion of swine manure affected by temperature and residual antibiotics, which will allow the development of an ARG elimination strategy before their release into the environment.

Design, synthesis, cytotoxicities and DNA cleavage activities of dibenzoxepine and isoquinoline derivatives starting from dehydroabietylamine.

A series of novel hexahydrodibenzoxepine and quinazoline derivatives were designed and synthesized starting from dehydroabietylamine. The cytotoxicities of the compounds against L02 and HepG2 cell lines were investigated. Meanwhile, the plasmid DNA (Escherichia coli) cleavage of several heterocyclic derivatives was studied. These compounds exhibit remarkable activities on plasmid DNA pBR322. Our study provides useful information for developing new and more potent antitumor agents.

Antitumor and scavenging radicals activities of some polyphenols related to dehydroabietylamine derivatives.

A novel series of polyphenols 4-9 were synthesized by the reaction of catechol with dehydroabietylamine derivatives. The antitumor activities of these compounds against L02 and HepG2 cells were investigated. Among them, compounds 4, 5, and 9 can inhibit HepG2 cells viability, but have lower inhibitory effect on L02 cells in the same concentration, indicating their potential for further development. Meanwhile, the novel series of polyphenols exhibited stronger radical-scavenging activities than the control groups.

A comparative study of antitumor activities and DNA cleavage on a class of dehydroabietylamine derivatives.

A series of novel dehydroabietylamine derivatives containing tricyclic diterpene structures were synthesized. The antitumor activities of these compounds against L02, Hey-1B and HepG2 cells were investigated. Significant activity was discovered forfourteen analogs. Meanwhile these compounds exhibit DNA cleavage activities on plasmid DNA (Escherichia coli), which depend on the Schiff base structure and the substituent of the aromatic moiety. Our findings present further information on the relationship between the chemical structure, biological function and DNA cleavage characteristics.

Study of oyster shell as a potential substrate for constructed wetlands.

We tested the suitability of oyster shell (OS) as a substrate for phosphorus removal in constructed wetlands (CWs) treating swine wastewater. OS is proven to have a significant phosphorus adsorption capacity; significant phosphorus removal was achieved in vertical subsurface flow constructed wetlands (VSSFs) that were filled with OS and used to treat swine wastewater. In the VSSF system, OS adsorption and precipitation played the greatest role in phosphorus removal, and the phosphorus distribution in the substrate layers was attributed to the vertical flow state of wastewater in the system. Ca-P was the predominant form of phosphorus in the system. Overall, the study results showed that OS could be used for phosphorus removal in CWs. OS also allowed for reuse of a waste substance, making the overall system more environmentally friendly.

Evaluation of rice straw and its transformation products on norfloxacin degradation and antibiotic resistome attenuation during soil incorporation.

Straw incorporation into reclaimed soils has been demonstrated to increase soil nutrients and has the potential to efficiently increase crop production. However, which incorporation mode is more helpful in the control of antibiotic resistance genes (ARGs) remains unknown. In this study, we systematically compared the occurrence of antibiotic resistome in norfloxacin contaminated soils amended with rice straw (RS) and the transformation products, biochar (RSB) and ash (RSA). RS significantly promoted the degradation of norfloxacin (0.0648 d-1, 3 times faster than control), whereas RSB had little effect and RSA hindered the degradation. Based on metagenomic analysis, RS and RSB significantly reduced the ARGs relative abundance (0.1421 and 0.1991 compared to 0.2540 in control) at the end of soil incubation. Adonis test indicated that all of amendment treatments significantly affect the microbial communities in soils, whereas only RS and RSB significantly affect the variation of antibiotic resistome. Procrustes analysis confirmed the association of microbial communities and ARGs. Network analysis further revealed that the reduction in Actinobacteria was the main reason for the general decrease of ARGs relative abundance during soil incorporation, whereas Proteobacteria and Bacteroidetes were responsible for temporary promotion of ARGs in RS and RSB at the early stage. Finally, scientifically setting up the usage of rice straw and optimizing the preparation process of biochar are suggested for the synchronous control of the risk of antibiotics and ARGs during soil incorporation.

Fitness reduction of antibiotic resistome by an extra carbon source during swine manure composting.

This study employed high-throughput quantitative polymerase chain reaction to evaluate the effects of specific co-substrate and additive on the fitness of antibiotic resistome during swine manure composting. The results showed that corncob particle as a co-substrate significantly reduced the relative abundances of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) simultaneously. The diversity of ARGs was also reduced more effectively by corncob addition. Brick granule as an additive reduced the concentrations of bioavailable Cu and Zn. However, the relative abundances of ARGs and MGEs were not reduced by the addition of brick granule subsequently. Redundancy analysis indicated a negative effect of the C content and positive effects of class I integrase gene (intI) and bioavailable metals on the variation of the relative abundance of ARGs (p < 0.01). The Procrustes test showed a higher goodness-of-fit between the relative abundance of ARGs and 16S rRNA genes (r = 0.8166; p < 0.0001). Our results suggests that the effect of corncob particle on the relative abundance of ARGs was achieved by driving the changes in physicochemical properties and microbial communities. This study confirmed the hypothesis of fitness cost and demonstrated the contribution of extra C source to ARG attenuation during composting.

Evaluation of wetland substrates for veterinary antibiotics pollution control in lab-scale systems.

The behaviors of typical veterinary antibiotics (oxytetracycline, ciprofloxacin and sulfamethazine) and 75 types of corresponding antibiotic resistant genes (ARGs) in four substrate systems (zeolite, gravel, red brick, and oyster shell) were investigated in this study. The results indicated that during treating synthetic livestock wastewater with individual antibiotic influent concentration of 100 µg/L, the effluent contained oxytetracycline and ciprofloxacin concentrations of 0.7-1.5 µg/L and 1.0-1.9 µg/L, respectively, in the zeolite and red brick systems, which were significantly lower than those of the other substrate systems (4.6-14.5 µg/L). Statistical correlation analyses indicated that the difference regarding oxytetracycline and ciprofloxacin removal among the four substrates was determined by their adsorption capacity which was controlled by the chemisorption mechanism. The average removal efficiency of sulfamethazine in the gravel system (48%) was higher than that of the other substrate systems (34-45%), and biodegradation may alter the sulfamethazine performance because of its co-metabolism process. Although tetG, floR, sul1, and qacEΔ1 were the dominant ARGs in all substrate systems (8.74 × 10-2-6.34 × 10-1), there was difference in the total ARG enrichment levels among the four substrates. Oyster shell exhibited the lowest total relative abundance (1.56 × 100) compared to that of the other substrates (1.82 × 100-2.27 × 100), and the ARG total relative abundance exhibited significant negative and positive correlations with the substrate pH and system bacterial diversity (P < 0.05), respectively. In summary, this study indicated that due to the difference of adsorption capacity and residual abundant nutrient in wastewater, the wetland substrate selection can affect the removal efficiency of veterinary antibiotics, and antibiotics may not be the determining factor of ARG enrichment in the substrate system.

Influence of hydraulic loading rate on antibiotics removal and antibiotic resistance expression in soil layer of constructed wetlands.

Behavior of veterinary antibiotics, the corresponding resistant genes in soil layer of constructed wetlands (red soil), and their response to different hydraulic loading rates (HLR) (2, 5, and 10 cm/d) were investigated. Results indicated that the soil layer had perfect performance for oxytetracycline and ciprofloxacin, yet sulfamethazine removal was unsatisfactory. Detection rates of oxytetracycline, ciprofloxacin and sulfamethazine in the effluent of simulation systems of soil layer were 8.33-36.36%, 8.33-47.83% and 100%, respectively. The model analysis of adsorption and hydrolysis indicated that physical adsorption, which was controlled by exchange reaction process based on diffusion, was the primary adsorption mechanism of target antibiotics in red soil, and the hydrolysis half-life values of antibiotics in the water of soil layer were shorter than them in wastewater. The removal response of oxytetracycline and ciprofloxacin to change of HLR was insignificant, yet the respective effluent concentrations of sulfamethazine at HLR of 2-10 cm/d were 41.90, 61.35 and 73.54 µg/L during treating synthetic livestock wastewater, which revealed significant positive correlation (P < 0.05). The relative abundances of each target resistance genes in soil showed significant increase after treating wastewater (10-5-10-6 to 10-4-10-1), and the total level of those at different HLRs (2, 5, and 10 cm/d) were 3.02 × 10-2, 7.54 × 10-2 and 8.65 × 10-1, respectively. In summary, HLR could affect the removal efficiency of partial antibiotic in soil layer of constructed wetlands, and the expression of antibiotic resistance in the soil gradually increased with increase in the HLR.

Niche separation and weak interactions in the high tidal zone of saltmarsh-mangrove mixing communities.

Saltmarsh-mangrove ecotones occur at the boundary of the natural geographic distribution of mangroves and salt marshes. Climate warming and species invasion can also drive the formation of saltmarsh-mangrove mixing communities. How these coastal species live together in a "new" mixed community is important in predicting the dynamic of saltmarsh-mangrove ecosystems as affected by ongoing climate change or human activities. To date, the understanding of species interactions has been rare on adult species in these ecotones.Two typical coastal wetlands were selected as cases to understand how mangrove and saltmarsh species living together in the ecotones. The leaves of seven species were sampled from these coastal wetlands based on their distribution patterns (living alone or coexisting) in the high tidal zone, and seven commonly used functional traits of these species were analyzed.We found niche separation between saltmarsh and mangrove species, which is probably due to the different adaptive strategies they adopted to deal with intertidal environments.Weak interactions between coexisting species were dominated in the high tidal zone of the two saltmarsh-mangrove communities, which could be driven by both niche differentiation and neutral theory.Synthesis. Our field study implies a potential opportunity to establish a multispecies community in the high tidal zone of saltmarsh-mangrove ecotones, where the sediment was characterized by low salinity and high nitrogen.

Synthesis, antitumor and DNA cleavage activities of a novel class of dehydroabietylamine derivatives.

Previous studies have reported higher biological activity of dehydrorosinamine derivatives. In order to further synthesize novel compounds with higher biological activity, a series of novel compounds containing benzo-azepine structures were synthesized from dehydroabietylamine in good yields in this study. The structures of synthesized compounds were identified by infra red (IR), 1H-NMR, 13C-NMR, and mass spectra (MS) analysis. The antitumor activities of the target compounds against L02 and HepG2 cells were studied. Furthermore, the dehydroabietylamine derivatives were studied on plasmid DNA cleavage activities. The results showed that the synthesized target compound exhibit antitumor and DNA cleavage activities against plasmid DNA (Escherichia coli). Our results further demonstrate the relationship between the chemical structure and biological function of the synthesized compounds.

Response of antibiotic resistance genes in constructed wetlands during treatment of livestock wastewater with different exogenous inducers: Antibiotic and antibiotic-resistant bacteria.

This work aimed to study the behavior of antibiotic resistance genes (ARGs) in constructed wetlands with different exogenous inducers additions (oxytetracycline and its resistant bacteria) by high-throughput quantitative polymerase chain reaction. Results indicated that constructed wetlands have the potential to reduce ARGs relative abundances in wastewater, and the total ARGs removal efficiency could exceed 60%. ARGs profile in the effluent differed from that in the influent, and that did not directly reflect the export of dominant ARGs in wetland biofilms. Meanwhile, the highest levels of detected numbers and relative abundances of ARGs were 43 and 3.35 × 10-1 for control system and 44 and 6.40 × 10-1 for treatment system, respectively, which meant that ARGs generation in wetlands were inevitable, and antibiotic and antibiotic-resistant bacteria from wastewater could indeed promote ARGs abundance in the system. Compared to the single roles of inducers, their synergistic role had a more significant influence on ARGs relative abundance.

[Pollution Characteristics and Risk Assessment of Antibiotics in Lianhua Reservoir].

Thirteen typical antibiotics in surface water of the Lianhua Reservoir were analyzed using HPLC/MS/MS to assess the pollution characteristics and risk levels. Ten antibiotics except for erythromycin, sulfadiazine, and sulfamethoxazole were detected in surface water and the total concentration of antibiotics varied between non-detectable (n.d.) and 925.26 ng ·L-1. Azithromycin had the highest concentration (n.d.-232.61 ng ·L-1) with the detection frequency of 75%, followed by enrofloxacin (n.d.-187.69 ng ·L-1), tetracycline (n.d.-155.05 ng ·L-1), and ciprofloxacin (n.d.-83.66 ng ·L-1) with the detection frequencies over 60%. The spatial distribution of antibiotics was as follows: total concentration of upstream (sampling point 1) > Aoxi River stream tributary (sampling point 2) > reservoir downstream (sampling point 3) > reservoir entrance (sampling point 4) > reservoir area (sampling point 5). The seasonal variations in the concentrations of antibiotics were evident; total concentrations in the dry season were significantly higher than those in the wet and normal seasons. The results of the environmental risk assessment indicated that ofloxacin, enrofloxacin, and ciprofloxacin pose significant risks to the environment. In the Lianhua Reservoir, ciprofloxacin showed high potential risk to the ecological environment, while the environmental risks of other antibiotics in the reservoir were below the medium level. The combined risk value of the antibiotics in the dry season was higher than that in the wet and normal seasons.

Effects of bulking material types on water consumption and pollutant degradation in composting process with controlled addition of different liquid manures.

This study was conducted to examine the effects of different bulking materials (corncob and ricehusk) on liquid manure consumption, organic matter degradation and pollutants retention in composting process under controlled addition of different types of liquid manures (LM). The results indicated that under the controlled addition of LM, bulking materials with higher content of biodegradable carbon (corncob) and LM with a higher concentration of organic pollutants (swine effluent) were more beneficial for biological heat generation and thus were more efficient for water evaporation, organic matter degradation, LM consumption and pollutants retention during the cocomposting process. Consequently, the optimization of these major influencing factors could compensate for efforts geared towards better utilization of the cocomposting process.

Effects of compost characteristics on nutrient retention and simultaneous pollutant immobilization and degradation during co-composting process.

This study was conducted to examine the effects of controlled addition of liquid (LM) to solid (SM) manure compost using a volume-model technique on the co-composting of SM and LM, and further to investigate the major effects of bulking material sizes and LM types on the co-composting process and final compost characteristics. Results indicated that this volume-model technique played a critical role in reducing leachate generation and improving the overall efficiency of the co-composting process. Specifically, the developed model enhanced the evaporation rates of windrows during the co-composting process. For improved final compost properties, small bulking materials and swine-effluent-based LM were found to be more efficient for organic matter degradation, LM consumption, hazardous metals immobilization, and essential nutrients retention than large bulking materials and biogas-based LM. Thus, process parameter optimizations represent major research options for successful co-composting applications for the future.

[Performance of an Aerobic Granular Reactor Treating Biogas Slurry from Pig Farm].

This research investigated the performance of an aerobic granular reactor treating biogas slurry from pig farm. Results indicated that the granular structure of aerobic sludge was not affected by the high pollution concentrationsin the biogas slurry. Although a low removal rate of phosphate was found in this study (about 16%±2%), organic matter and ammonia nitrogen showed stable removal and transformation in the granular system, and the effluent concentrations of those components were (267±81)mg·L-1 and(62±12)mg·L-1, respectively. In addition, the removal rate of sulfamethazine and tetracycline was 98%±2% and 65%±16%, respectively. During the process biogas slurry treatment, bacterial communities in the aerobic granular reactor remained stable, and Comamonadaceae was the dominant bacteria (relative abundance ofapproximately 16.66%).

Biodiesel production from microbial granules in sequencing batch reactor.

Effect of reaction variables of in situ transesterification on the biodiesel production, and the characteristic differences of biodiesel obtained from aerobic granular sludge (AG) and algae-bacteria granular consortia (AAG) were investigated. The results indicated that the effect of variables on the biodiesel yield decreased in the order of methanol quantity > catalyst concentration > reaction time, yet the parameters change will not significantly affect biodiesel properties. The maximum biodiesel yield of AAG was 66.21 ±â€¯1.08 mg/g SS, what is significant higher than that of AG (35.44 ±â€¯0.92 mg/g SS). Although methyl palmitate was the dominated composition of biodiesel obtained from both granules, poly-unsaturated fatty acid in the AAG showed a higher percentage (21.86%) than AG (1.2%) due to Scenedesmus addition. Further, microbial analysis confirmed that the composition of biodiesel obtained from microbial granules was also determined by bacterial community, and Xanthomonadaceae and Rhodobacteraceae were the dominant bacteria of AG and AAG, respectively.