Agricultural land-use change exacerbates the dissemination of antibiotic resistance genes via surface runoffs in Lake Tai Basin, China.

Agricultural runoff is an important antibiotic resistance genes (ARGs) dissemination pathway from farmlands to water environment, however few studies have focused on the influence of agricultural land-use change on the pattern of ARGs in runoff and assess the health risk to public. Lake Tai Basin which experiences agricultural land-use change was selected to elucidate this concern. Our findings revealed that the pattern of ARGs was more diverse and the gene abundance was higher in orchard runoffs by comparison with conventional cropland runoffs. Co-occurrence network analysis between mobile genetic elements and ARGs demonstrated that after agricultural land-use change, ARG dissemination via runoffs became more threatened. In addition, this study illustrated the correlations between the antibiotic resistome and microbiome in runoffs, finding that non-dominant microbial taxa were the limiting factor which determined the pattern of ARGs in surface runoffs. In summary, the pattern and dissemination risk of ARGs in the surface runoff after agricultural land-use change in Lake Tai Basin were clarified via this study.

Stable-isotope probing of bacterial community for dissolved inorganic carbon utilization in Microcystis aeruginosa-dominated eutrophic water.

Dissolved inorganic carbon (DIC) is an important source of carbon in aquatic ecosystems, especially under conditions of increased frequency of cyanobacterial bloom. However, the importance of bacteria in direct or indirect utilization of DIC has been widely overlooked in eutrophic freshwater. To identify the functional bacteria that can actively utilize DIC in eutrophic freshwater during cyanobacterial bloom, stable-isotope probing (SIP) experiments were conducted on eutrophic river water with or without inoculation with cyanobacteria (Microcystis aeruginosa). Our 16S rRNA sequencing results revealed the significance of Betaproteobacteria, with similar relative abundance as Alphaproteobacteria, in the active assimilation of H13CO3- into their DNA directly or indirectly, which include autotrophic genera Betaproteobacterial ammonia-oxidizing bacteria. Other bacterial groups containing autotrophic members, e.g. Planctomycetes and Nitrospira, also presented higher abundance among free-living bacteria in water without cyanobacteria. Microcystis aggregates showed a preference for some specific bacterial members that may utilize H13CO3- metabolized by Microcystis as organic matter, e.g. Bacteroidetes (Cytophagales, Sphingobacteriales), and microcystin-degrading bacteria Betaproteobacteria (Paucibacter/Burkholderiaceae). This study provides some valuable information regarding the functional bacteria that can actively utilize DIC in eutrophic freshwater.

Soil anammox community structure in different land use soils treatment with 13C urea as determined by analysis of phospholipid fatty acids.

The anaerobic ammonium oxidation (anammox) process is globally an important nitrogen-cycling process mediated by specialized microbes. However, still little information is documented about anammox microbial community structure under agricultural soils. The anaerobic incubation experiment was conducted to study the impacts of different land use soils fertilized by 13C-urea on the activity and diversity of anammox bacteria using stable isotope to probe the phospholipid fatty acid (PLFA-SIP). The 13C was preferentially incorporated in ratios PLFAs 16:1ω7c, 16:1ω5c, and 16:0. The results revealed that the abundance of the anammox bacteria (both hzs-ß and hzo) were observed in vegetable soil V1 and paddy soils (R1 and R2) means that they were positively correlated with 13C-urea but were negatively correlated with NO3--N and NH4+-N concentrations. Thus, 13C-PLFAs 16:1ω7c, 16:1ω5c, and 16:0 could be the biomarker as soil anammox. The anaerobic microbial community composition of soils under different land use systems was diverse, and V1, R1, and R2 had similar microbial diversity and higher microbial biomass. The principal component analysis between soil properties and gene abundance suggested that not only pH but also soil organic matter, available P, and available K were important factors for the anammox process. This study suggested that 13C-Urea-PLFA for anaerobic incubation was a simple method to study anammox microbial community structure through affecting the soil nutrients, and the different land use systems played important roles in determining the microbial composition of soils.

Sulfur reduces the root-to-shoot translocation of arsenic and cadmium by regulating their vacuolar sequestration in wheat (Triticum aestivum L.).

Accumulation of arsenic (As) and cadmium (Cd) in wheat grain is a serious threat to human health. Sulfur (S) can simultaneously decrease wheat grain As and Cd concentrations by decreasing their translocation in wheat; however, the mechanisms are unclear. We conducted hydroponic experiments to explore the mechanisms by which S modulates As and Cd translocation and their toxicity in wheat. Wheat seedlings were grown in deficient sulfate (2.5 µM) or sufficient sulfate (1.0 mM) nutrient solutions for 6 days and then exposed to zero (control), low As+Cd (1 µM As plus 0.5 µM Cd), or high As+Cd (50 µM As plus 30 µM Cd) for another 6 days. Compared with the control, plant growth was not affected by low As+Cd, but was significantly inhibited by high As+Cd. In the low As+Cd treatment, S supply had no significant effect on plant growth or root-to-shoot As and Cd translocation. In the high As+Cd treatment, sufficient S supply significantly alleviated As and Cd toxicity and their translocation by increasing phytochelatin (PC) synthesis and the subsequent vacuolar sequestration of As and Cd in roots, compared with deficient S supply. The use of L-buthionine sulfoximine (a specific inhibitor of γ-glutamylcysteine synthetase) confirmed that the alleviation of As and Cd translocation and toxicity in wheat by S is mediated by increased PC production. Also, TaHMA3 gene expression in wheat root was not affected by the As+Cd and S treatments, but the expression of TaABCC1 was upregulated by the high As+Cd treatment and further increased by sufficient S supply and high As+Cd treatment. These results indicate that S-induced As and Cd subcellular changes affect As and Cd translocation mainly by regulating thiol metabolism and ABCC1 expression in wheat under As and Cd stress.

Agricultural land-use change and rotation system exert considerable influences on the soil antibiotic resistome in Lake Tai Basin.

In this study, we use high-throughput quantitative polymerase chain reaction approaches to comprehensively assess the effects of agricultural land-use change on the antibiotic resistome of agricultural runoffs after rainfalls in Lake Tai Basin. For the first time in this region, our findings show that orchard runoffs harbored more diverse and abundant antibiotic resistance genes (ARGs) than traditional cropland runoffs. Network analysis demonstrated that orchard runoffs possessed a strong ability for ARG dissemination via horizontal gene transfer. These results suggest that residents might be exposed to a higher public health threat than before. Moreover, the present study confirmed that the rice-wheat rotation system plays a key role in regulating the soil antibiotic resistome profile. Using 16S rRNA high-throughput sequencing technology, this study clarified the relationships between the antibiotic resistome and soil microbiome composition. Finally, we discuss the key environmental factors driving changes in the soil antibiotic resistome. In summary, this study gives insight into the dissemination of environmental ARGs to the people living in the Lake Tai Basin.

The effect of organic solvent washing on the structure of hydrochar-based dissolved organic matters and its potential environmental toxicity.

With the increased interest in the practical use of hydrochar, concerns about the possible environmental biotoxicity of hydrochar and its released dissolved organic matters (DOM) have grown. As a common method for removing bio-oil on the surface of hydrochar, the effect of organic solvent washing on the properties of hydrochar released DOM remains unclear. In this study, we made a comprehensive comparison of hydrochar properties and molecule structure as well as biotoxicity of DOM released from HC (raw hydrochar) and THC (hydrochar washed by tetrahydrofuran). The results indicated that the mass loss of hydrochar was obvious after tetrahydrofuran (THF) washing, and a decline of H/C atomic ratio and increase of N/C and O/C atomic ratios was observed based on Van Krevelen (VK) diagram. This result was further confirmed by FTIR, 13C NMR, and XPS results. Meanwhile, the molecule structure of DOM was shifted to lower molecule weight with higher O-contain compounds after THF extraction due to the demethanation process. However, the biotoxicity experiments indicated that both extracted DOM had no significant impact on germination rate of wheat, and HC-treated sample even exhibited growth superiority. Nevertheless, potential toxicity was observed with the increase of the activity of antioxidant enzymes, and THF washing aggravated the potential oxidative damage through increasing the aromaticity of DOM. Such understanding highlights the importance of evaluating hydrochar and its released DOM before applications, so as to reduce the potential environment biotoxicity.

Straw-derived biochar mitigates CO2 emission through changes in soil pore structure in a wheat-rice rotation system.

To better understand the relationships between soil pore structure features and soil CO2 emission and soil organic carbon (SOC) sequestration following different straw return modes, undisturbed soil cores (0-5 cm and 5-10 cm) were collected from a rice-wheat rotation system under 4 straw return treatments as (1) no straw return (CK), (2) straw direct return (DR), (3) straw biochar return (BR); (4) straw-pig manure fermentation return (FR) for six years. Pore structure parameters including pore size distribution, porosity, connectivity, anisotropy and fractal dimension (FD) were determined using X-ray computer tomography. Soil CO2 flux and concentrations of SOC, readily oxidable carbon and nutrients were also measured. The results showed that BR and FR had significantly higher SOC concentration than DR and CK. Porosity and number of >500 µm and 500-100 µm macropores, FD and connectivity were significantly highest under FR and was lowest under BR. FR and DR produced 28.1%-32.4% higher C-CO2 than CK and BR in wheat growing season, and 9.80%-16.9% higher in rice season. Soil CO2 emission and C concentrations were significantly related to soil pore structure parameters. The CO2 emission was most significantly related to number of >500 µm pores and FD, indicating that poorly developed pore structure under BR hindered the production and diffusion of CO2 from soil. These results enhanced our understanding of the relationship between soil pore structure and CO2 emission following biochar application, and provided evidence for decision making process in choosing proper straw managements to promote SOC sequestration and reduce CO2 emission.

Interactions between elevated CO2 levels and floating aquatic plants on the alteration of bacterial function in carbon assimilation and decomposition in eutrophic waters.

Elevated atmospheric CO2 concentration (eCO2) may have different effects on the bacterial community with regard to C assimilation and decomposition in eutrophic waters compared to that in fresh waters with intermediate levels of nutrients and oceans. Aquatic plant growth under eCO2 could further modify microbial activities associated with the C cycle in eutrophic waters. Therefore, there is an urgent need to further study how eCO2 and its interactions with the growth of aquatic plants affect the composition and function of the bacterial community involved in mediating the C cycle in eutrophic waters. Accordingly, we designed a microcosm experiment to investigate the effects of ambient and high CO2 concentrations on bacterial community composition and function in eutrophic waters with and without the growth of Eichhornia crassipes (Mart.) Solms. The results from 16S rRNA gene sequencing, function prediction, and q-PCR showed that eCO2 significantly increased the abundance of bacterial and functional genes involved in CO2 assimilation (photosynthetic bacteria; cbbL IA & IC, cbbL ID, cbbM, pufM) and C decomposition (Acidimicrobiia, Thermoleophilia, Gaiellales; ChiA), illustrating the functional enrichment with photoautotrophy, hydrocarbon degradation, cellulolysis, and aromatic hydrocarbon degradation. However, eCO2 decreased the abundance of some chemoautotrophic bacteria, including nitrifying bacteria (Nitrospirae, Nitrosomonadaceae). In contrast, the cultivation of E. crassipes decreased the abundance of photosynthetic bacteria but increased the abundance of bacteria involved in complex C decomposition associated with root exudates and degradation, e.g. Fibrobacteres, Sphingobacteriales, Sphingomonadales, and Rhizobiales. eCO2 and growth of E. crassipes had opposite effects on algal density in eutrophic waters, creating interactive effects that further decreased the diversity of the bacterial community and abundance of some CO2-assimilating bacteria with nitrifying characteristics (Nitrosomonadaceae) and some C-degrading bacteria (Fibrobacteres) with denitrifying properties (Flavobacteriaceae, Sphingomonadaceae, and Gemmobacter). Therefore, the interactions between aquatic plants and the bacterial community in eutrophic waters under eCO2 would be beneficial to the environment and help alleviate the greenhouse effect.

Comprehensive evaluation of a cost-effective method of culturing Chlorella pyrenoidosa with unsterilized piggery wastewater for biofuel production.

BACKGROUND: The utilization of Chlorella for the dual goals of biofuel production and wastewater nutrient removal is highly attractive. Moreover, this technology combined with flue gas (rich in CO2) cleaning is considered to be an effective way of improving biofuel production. However, the sterilization of wastewater is an energy-consuming step. This study aimed to comprehensively evaluate a cost-effective method of culturing Chlorella pyrenoidosa in unsterilized piggery wastewater for biofuel production by sparging air or simulated flue gas, including algal biomass production, lipid production, nutrient removal rate and the mutual effects between algae and other microbes. RESULTS: The average biomass productivity of C. pyrenoidosa reached 0.11 g L-1 day-1/0.15 g L-1 day-1 and the average lipid productivity reached 19.3 mg L-1 day-1/30.0 mg L-1 day-1 when sparging air or simulated flue gas, respectively. This method achieved fairish nutrient removal efficiency with respect to chemical oxygen demand (43.9%/55.1% when sparging air and simulated flue gas, respectively), ammonia (98.7%/100% when sparging air and simulated flue gas, respectively), total nitrogen (38.6%/51.9% when sparging air or simulated flue gas, respectively) and total phosphorus (42.8%/60.5% when sparging air or simulated flue gas, respectively). Culturing C. pyrenoidosa strongly influenced the microbial community in piggery wastewater. In particular, culturing C. pyrenoidosa enriched the abundance of the obligate parasite Vampirovibrionales, which can result in the death of Chlorella. CONCLUSION: The study provided a comprehensive evaluation of culturing C. pyrenoidosa in unsterilized piggery wastewater for biofuel production. The results indicated that this cost-effective method is feasible but has considerable room for improving. More importantly, this study elucidated the mutual effects between algae and other microbes. In particular, a detrimental effect of the obligate parasite Vampirovibrionales on algal biomass and lipid production was found.

Contrasting impact of elevated atmospheric CO2 on nitrogen cycle in eutrophic water with or without Eichhornia crassipes (Mart.) Solms.

The elevation of atmospheric CO2 is an inevitable trend that would lead to significant impact on the interrelated carbon and nitrogen cycles through microbial activities in the aquatic ecosystem. Eutrophication has become a common trophic state of inland waters throughout the world, but how the elevated CO2 affects N cycles in such eutrophic water with algal bloom, and how vegetative restoration helps to mitigate N2O emission remains unknown. We conducted the experiments to investigate the effects of ambient and elevated atmospheric CO2 (a[CO2], e[CO2]; 400, 800 µmol﹒mol-1) with and without the floating aquatic plant, Eichhornia crassipes (Mart.) Solms, on N-transformation in eutrophic water using the 15N tracer method. The nitrification could be slightly inhibited by e[CO2], due mainly to the competition for dissolved inorganic carbon between algae and nitrifiers. The e[CO2] promoted denitrification and N2O emissions from eutrophic water without growth of plants, leading to aggravation of greenhouse effect and forming a vicious cycle. However, growth of the aquatic plant, Eichhornia crassipes, slightly promoted nitrification, but reduced N2O emissions from eutrophic water under e[CO2] conditions, thereby attenuating the negative effect of e[CO2] on N2O emissions. In the experiment, the N transformation was influenced by many factors such as pH, DO and algae density, except e[CO2] and plant presence. The pH could be regulated through diurnal photosynthesis and respiration of algae and mitigated the acidification of water caused by e[CO2], leading to an appropriate pH range for both nitrifying and denitrifying microbes. Algal respiration at night could consume DO and enhance abundance of denitrifying functional genes (nirK, nosZ) in water, which was also supposed to be a critical factor affecting denitrification and N2O emissions. This study clarifies how the greenhouse effect caused by e[CO2] mediates N biogeochemical cycle in the aquatic ecosystem, and how vegetative restoration mitigates greenhouse gas emission.