Markedly Increasing Antibiotic Resistance and Dual Treatment of Neisseria gonorrhoeae Isolates in Guangdong, China, from 2013 to 2020.

The emergence of multidrug resistance in Neisseria gonorrhoeae is concerning, especially the cooccurrence of azithromycin resistance and decreased susceptibility to extended-spectrum cephalosporin. This study aimed to confirm the antibiotic resistance trends and provide a solution for N. gonorrhoeae treatment in Guangdong, China. A total of 5,808 strains were collected for assessment of antibiotic MICs. High resistance to penicillin (53.80 to 82%), tetracycline (88.30 to 100%), ciprofloxacin (96 to 99.8%), cefixime (6.81 to 46%), and azithromycin (8.60 to 20.03%) was observed. Remarkably, spectinomycin and ceftriaxone seemed to be the effective choices, with resistance rates of 0 to 7.63% and 2.00 to 16.18%, respectively. Moreover, the rates of azithromycin resistance combined with decreased susceptibility to ceftriaxone and cefixime reached 9.28% and 8.64%, respectively. Furthermore, genotyping identified NG-STAR-ST501, NG-MAST-ST2268, and MLST-ST7363 as the sequence types among representative multidrug-resistant isolates. Evolutionary analysis showed that FC428-related clones have spread to Guangdong, China, which might be a cause of the rapid increase in extended-spectrum cephalosporin resistance currently. Among these strains, the prevalence of N. gonorrhoeae was extremely high, and single-dose ceftriaxone treatment might be a challenge in the future. To partially relieve the treatment pressure, a susceptibility test for susceptibility to azithromycin plus extended-spectrum cephalosporin dual therapy was performed. The results showed that all the representative isolates could be effectively killed with the coadministration of less than 1 mg/liter azithromycin and 0.125 mg/liter extended-spectrum cephalosporin, with a synergistic effect according to a fractional inhibitory concentration (FIC) of <0.5. In conclusion, dual therapy might be a powerful measure to treat refractory N. gonorrhoeae in the context of increasing antibiotic resistance in Guangdong, China.

Impact of influent strengths on nitrous oxide emission and its molecular mechanism in constructed wetlands treating swine wastewater.

Constructed wetlands (CWs) can remove nitrogen (N) through plant assimilation and microbial nitrification and denitrification, while it also releases large greenhouse gas nitrous oxide (N2O) into the atmosphere. However, N2O emissions and the underlying microbial mechanisms of CWs when treating high-strength wastewater have not been systematically surveyed. Here, the effect of three influent strengths on N2O emissions in a pilot-scale CW treatment of swine wastewater was determined and the underlying microbial mechanisms were explored. The results showed that the removal rates of ammonium (NH4+) and total nitrogen (TN) increased significantly with the increasing influent strengths, however, the ratio of N2O emission/TN removal rose by 1.5 times at the same time. Quantitation of microorganisms responsible for N-cycle in the sediment indicated that the abundance of ammonia-oxidizing bacteria (AOB) in high influent strengths (COD, 962.38 ± 3.05 mg/L; NH4+, 317.89 ± 4.24 mg/L) was 51.6-fold compared with that in low influent strengths (COD, 516.94 ± 4.18 mg/L; NH4+, 100.65 ± 2.65), and AOB gradually replaced ammonia-oxidizing archaea (AOA) to dominate ammonia oxidizers. Structural equation models demonstrated that NO2- accumulations promoted the ratio of AOB/AOA, which further led to an increase in the ratio of N2O emission/TN removal. It is worth noting both the N removal rates and N2O emissions increased with the increasing influent strength. To obtain reduced N2O emissions, pretreatment technology for strength reduction should be supplemented before high-strength wastewater enters the CWs. This study may shed new light on the sustainable operation and application of CWs.

Ecological selection of bacterial taxa with larger genome sizes in response to polycyclic aromatic hydrocarbons stress.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous priority pollutants that cause great damage to the natural environment and health. Average genome size in a community is critical for shedding light on microbiome's functional response to pollution stress within an environment. Here, microcosms under different concentrations were performed to evaluate the selection of PAHs stress on the average genome size in a community. We found the distinct communities of significantly larger genome size with the increase of PAHs concentration gradients in soils, and consistent trends were discovered in soils at different latitudes. The abundance of Proteobacteria and Deinococcus-Thermus with relatively larger genomes increased along with PAHs stress and well adapted to polluted environments. In contrast, the abundance of Patescibacteria with a highly streamlined and smaller genome decreased, implying complex interactions between environmental selection and functional fitness resulted in bacteria with larger genomes becoming more abundant. Moreover, we confirmed the increased capacity for horizontal transfer of degrading genes between communities by showing an increased connection number per node positively related to the nidA gene along the concentration gradients in the co-occurrence network. Our findings suggest PAHs tend to select bacterial taxa with larger genome sizes, with significant consequences for community stability and potential biodegradation strategies.

Anaerobic degradation of xenobiotic organic contaminants (XOCs): The role of electron flow and potential enhancing strategies.

In groundwater, deep soil layer, sediment, the widespread of xenobiotic organic contaminants (XOCs) have been leading to the concern of human health and eco-environment safety, which calls for a better understanding on the fate and remediation of XOCs in anoxic matrices. In the absence of oxygen, bacteria utilize various oxidized substances, e.g. nitrate, sulphate, metallic (hydr)oxides, humic substance, as terminal electron acceptors (TEAs) to fuel anaerobic XOCs degradation. Although there have been increasing anaerobic biodegradation studies focusing on species identification, degrading pathways, community dynamics, systematic reviews on the underlying mechanism of anaerobic contaminants removal from the perspective of electron flow are limited. In this review, we provide the insight on anaerobic biodegradation from electrons aspect - electron production, transport, and consumption. The mechanism of the coupling between TEAs reduction and pollutants degradation is deconstructed in the level of community, pure culture, and cellular biochemistry. Hereby, relevant strategies to promote anaerobic biodegradation are proposed for guiding to an efficient XOCs bioremediation.

Effective removal of antibiotic resistance genes and potential links with archaeal communities during vacuum-type composting and positive-pressure composting.

As a major reservoir of antibiotics, animal manure contributes a lot to the augmented environmental pressure of antibiotic resistance genes (ARGs). This might be the first study to explore the effects of different ventilation types on the control of ARGs and to identify the relationships between archaeal communities and ARGs during the composting of dairy manure. Several ARGs were quantified via Real-time qPCR and microbial communities including bacteria and archaea were analyzed by High-throughput sequencing during vacuum-type composting (VTC) and positive-pressure composting (PPC). The total detected ARGs and class I integrase gene (intI1) under VTC were significantly lower than that under PPC during each stage of the composting (p<0.001). The relative abundance of potential human pathogenic bacteria (HPB) which were identified based on sequencing information and correlation analysis decreased by 74.6% and 91.4% at the end of PPC and VTC, respectively. The composition of archaeal communities indicated that methane-producing archaea including Methanobrevibacter, Methanocorpusculum and Methanosphaera were dominant throughout the composting. Redundancy analysis suggested that Methanobrevibacter and Methanocorpusculum were positively correlated with all of the detected ARGs. Network analysis determined that the possible hosts of ARGs were different under VTC and PPC, and provided new sights about potential links between archaea and ARGs. Our results showed better performance of VTC in reducing ARGs and potential HPB and demonstrated that some archaea could also be influential hosts of ARGs, and caution the risks of archaea carrying ARGs.

Genetic diversity of diazotrophs and total bacteria in the phyllosphere of Pyrus serotina, Prunus armeniaca, Prunus avium, and Vitis vinifera.

The phyllosphere, which supports a large number of microorganisms, represents the interface between the aboveground parts of plants and air. In this study, four nifH clone libraries were constructed from the phyllosphere of Pyrus serotina (L), Vitis vinifera (P), Prunus armeniaca (X), and Prunus avium (Y). Clones related to Skermanella (L, 12.1%; X, 15.6%; Y, 62.5%; P 70.8%), Bradyrhizobium (X, 2.1%; P, 15.1%; L, 63.7%), Erwinia (X, 68.8%), Pseudomonas (L, 3.3%; P, 7.6%), and Chroococcidiopsis (P, 0.9%; L, 4.4%, X; 5.2%, Y; 19.6%) were present at high percentages, highlighting their critical role in contributing nitrogen to the phyllosphere ecosystem. The 16S rDNA sequence analysis suggested that phyllosphere-associated bacteria were affiliated with a wide range of taxa, encompassing members from Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Cyanobacteria, Tenericutes, and Deinococcus-Thermus. Additionally, the abundance of the nifH gene and 16S rDNA was assessed with quantitative PCR. The number of copies of nifH and 16S rDNA ranged from 1.14 × 103 to 1.49 × 104 and from 3.72 × 106 to 7.02 × 107 copies/g fresh leaf sample, respectively. In conclusion, our work sheds light on the microbial communities of the phyllosphere that are important for plant growth. Moreover, we observed a unique composition of nitrogen-fixing bacteria in each phyllosphere sample, suggesting the existence of specific interactions between these functional microorganism and plants, which may provide information or be a reference for the development of bacterial fertilizers.

3D Printing of BaTiO3 Piezoelectric Ceramics for a Focused Ultrasonic Array.

BaTiO3 (BTO) ceramics were fabricated based on stereolithography technology. The microstructures and electric properties of the BTO ceramics were studied. X-ray patterns of sintered BTO ceramics indicated that the tetragonal phase had formed, and the grain size increased clearly as BTO weight percentage increased. Moreover, the BTO ceramics exhibited good electric properties, with a piezoelectric constant d33 of 166 pC/N at 80% BTO weight percentage. To evaluate the properties of 3D printed BTO ceramics, a 1.4 MHz focused ultrasonic array was fabricated and characterized. The -6dB bandwidth of the array was 40%, and the insertion loss at the center frequency was 50 dB. The results show that the printed BTO ceramics array have good potential to be used in ultrasonic transducers for various applications.

Enhancement of facultative anaerobic denitrifying communities by oxygen release from roots of the macrophyte in constructed wetlands.

The radial oxygen loss (ROL) of wetland plants is a crucial factor that can influence the efficiency required for nitrogen (N) removal and microbial activities responsible for N removal in constructed wetlands (CWs). However, the shift of microbial community in different niches in response to ROL has been rarely studied. This study aims to unravel the link between the ROL and microbial response in sediment, water and rhizoplane by a surface flow CW planted with Myriophyllum aquaticum for treating high-strength swine wastewater. Ti3+-citrate colorimetric method demonstrated that M. aquaticum was a wetland species with a ROL of 0.019 mg/h/plant. Using quantitative polymerase chain reactions (qPCR) and high-throughput sequencing of microbial 16S rRNA gene, we demonstrated that the abundance of facultative anaerobic denitrifiers in the rhizoplane was the most of the three niches, that in the water (5-10 cm) was the less and that in the sediment was the least. Acinetobacter was enriched and dominated amongst denitrifiers in the water. Denitrifiers in the rhizoplane were mainly dominated by enriched Pseudomonas, Aeromonas, and Acinetobacter. The theoretical calculation of oxygen sources and consumptions indicated that water reaeration should support the oxygen demands for nitrification in the aerobic layer (0-5 cm), and the ROL could stimulate the growth of facultative anaerobic denitrifiers in the rhizoplane and water (5-10 cm) to achieve denitrification within CW systems.

Mitigation of nitrous oxide emissions from acidic soils by Bacillus amyloliquefaciens, a plant growth-promoting bacterium.

Nitrous oxide (N2 O) is a long-lived greenhouse gas that can result in the alteration of atmospheric chemistry and cause accompanying changes in global climate. To date, many techniques have been used to mitigate the emissions of N2 O from agricultural fields, which represent one of the most important sources of N2 O. In this study, we designed a greenhouse pot experiment and a microcosmic serum bottle incubation experiment using acidic soil from a vegetable farm to study the effects of Bacillus amyloliquefaciens (BA) on plant growth and N2 O emission rates. The addition of BA to the soil promoted plant growth enhanced the soil pH and increased the total nitrogen (TN) contents in the plants. At the same time, it decreased the concentrations of ammonium (NH4+ ), nitrate (NO3- ) and TN in the soil. Overall, the addition of BA resulted in a 50% net reduction of N2 O emissions compared with the control. Based on quantitative PCR and the network analysis of DNA sequencing, it was demonstrated that BA partially inhibited the nitrification process through the significant reduction of ammonia oxidizing bacteria. Meanwhile, it enhanced the denitrification process, mainly by increasing the abundance of N2 O-reducing bacteria in the treatment with BA. The results of our microcosm experiment provided evidence that strongly supported the above findings under more strictly controlled laboratory conditions. Taken together, the results of our study evidently demonstrated that BA has dual effects on the promotion of plant growth and the dramatic reduction of greenhouse emissions, thus suggesting the possibility of screening beneficial microbial organisms from the environment that can promote plant growth and mitigate greenhouse trace gases.

Linking N2O Emissions from Biofertilizer-Amended Soil of Tea Plantations to the Abundance and Structure of N2O-Reducing Microbial Communities.

Nitrous oxide (N2O) contributes up to 8% of global greenhouse gas emissions, with approximately 70% from terrestrial sources; over one-third of this terrestrial emission has been linked to increased agricultural fertilizer use. Much of the nitrogen in fertilizers is converted to N2O by microbial processes in soil. However, the potential mechanism of biofertilizers and the role of microbial communities in mitigating soil N2O emissions are not fully understood. Here, we used a greenhouse-based pot experiment with tea plantation soil to investigate the effect of Trichoderma viride biofertilizer on N2O emission. The addition of biofertilizer reduced N2O emissions from fertilized soil by 67.6%. Quantitative PCR (qPCR) analysis of key functional genes involved in N2O generation and reduction ( amoA, nirK, nirS, and nosZ) showed an increased abundance of nirS and nosZ genes linked to the pronounced reduction in N2O emissions. High-throughput sequencing of nosZ showed enhanced relative abundance of nosZ-harboring denitrifiers in the T. viride biofertilizer treatments, thus linking greater N2O reduction capacity to the reduced emissions. Our findings showed that biofertilizers can affect the microbial nitrogen transformation process and reduce N2O emissions from agroecosystems.

Effect of Trichoderma viride biofertilizer on ammonia volatilization from an alkaline soil in Northern China.

Ammonia (NH3) volatilization is one of the primary pathways of nitrogen (N) loss from soils after chemical fertilizer is applied, especially from the alkaline soils in Northern China, which results in lower efficiency for chemical fertilizers. Therefore, we conducted an incubation experiment using an alkaline soil from Tianjin (pH8.37-8.43) to evaluate the suppression effect of Trichoderma viride (T. viride) biofertilizer on NH3 volatilization, and compared the differences in microbial community structure among all samples. The results showed that viable T. viride biofertilizer (T) decreased NH3 volatilization by 42.21% compared with conventional fertilizer ((CK), urea), while nonviable T. viride biofertilizer (TS) decreased NH3 volatilization by 32.42%. NH3 volatilization was significantly higher in CK and sweet potato starch wastewater (SPSW) treatments during the peak period. T. viride biofertilizer also improved the transfer of ammonium from soil to sweet sorghum. Plant dry weights increased 91.23% and 61.08% for T and TS, respectively, compared to CK. Moreover, T. viride biofertilizer enhanced nitrification by increasing the abundance of ammonium-oxidizing archaea (AOA) and ammonium-oxidizing bacteria (AOB). The results of high-throughput sequencing indicated that the microbial community structure and composition were significantly changed by the application of T. viride biofertilizer. This study demonstrated the immense potential of T. viride biofertilizer in reducing NH3 volatilization from alkaline soil and simultaneously improving the utilization of fertilizer N by sweet sorghum.

Electron Transfer in the Biogeochemical Sulfur Cycle.

Microorganisms are key players in the global biogeochemical sulfur cycle. Among them, some have garnered particular attention due to their electrical activity and ability to perform extracellular electron transfer. A growing body of research has highlighted their extensive phylogenetic and metabolic diversity, revealing their crucial roles in ecological processes. In this review, we delve into the electron transfer process between sulfate-reducing bacteria and anaerobic alkane-oxidizing archaea, which facilitates growth within syntrophic communities. Furthermore, we review the phenomenon of long-distance electron transfer and potential extracellular electron transfer in multicellular filamentous sulfur-oxidizing bacteria. These bacteria, with their vast application prospects and ecological significance, play a pivotal role in various ecological processes. Subsequently, we discuss the important role of the pili/cytochrome for electron transfer and presented cutting-edge approaches for exploring and studying electroactive microorganisms. This review provides a comprehensive overview of electroactive microorganisms participating in the biogeochemical sulfur cycle. By examining their electron transfer mechanisms, and the potential ecological and applied implications, we offer novel insights into microbial sulfur metabolism, thereby advancing applications in the development of sustainable bioelectronics materials and bioremediation technologies.

Non-negligible roles of upstream rivers in determining the antibiotic resistance genes community in an interconnected river-lake system (Dongting lake, China).

Emergence and spread of antibiotic resistance genes (ARGs) in lakes have been considered as a global health threat. However, a thorough understanding of the distribution patterns and ecological processes that shape the ARGs profile in interconnected river-lake systems remains largely unexplored. In this study, we collected paired water and sediment samples from a typical interconnected river-lake system, Dongting Lake in China, during both wet and dry seasons. Using high-throughput quantitative PCR, we investigated the spatial and temporal distribution of ARGs and the factors that influence them. A total of 8 major antibiotic classes and 10 mobile genetic elements were detected across the Dongting Lake basin. The unique hydrological characteristics of this interconnected river-lake system result in a relatively stable abundance of ARGs across different seasons and interfaces. During the wet season, deterministic processes dominated the assembly of ARGs, allowing environmental factors, such as heavy metals, to serve as main driving forces of ARGs distribution. When the dry season arrived, variations in hydrological conditions and changes in ARGs sources caused stochastic processes to dominate the assembly of ARGs. Our findings provide valuable insights for understanding the ecological processes of ARGs in interconnected river-lake systems, emphasizing the necessity of upstream restoration and clarifying river-lake relationships to mitigate ARGs dissemination.

Transfer and accumulation of antibiotic resistance genes and bacterial pathogens in the mice gut due to consumption of organic foods.

Over the last few decades, organic food demand has grown largely because of increasing personal health concerns. Organic farming introduces antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB) into foods. However, potential effects of organic foods on the gut microbiome and ARGs have been overlooked. Using high-throughput quantitative PCR and 16S rRNA high-throughput sequencing technology, we examined 132 ARGs from major classes, eight transposase genes, universal class I integron-integrase gene (intI), clinical class I integron-integrase gene (cintI), and the bacterial community in mouse gut after 8 weeks with an either organic or inorganic lettuce and wheat diet. A total of 8 types of major ARGs and 10 mobile genetic elements (MGEs) were detected in mice gut, including tetracycline, multidrug, sulfonamide, aminoglycoside, beta-lactamase, chloramphenicol, MLSB and vancomycin resistance genes. We found that abundance and diversity of ARGs, mobile gene elements, and potential ARB in the gut increased with time after consumption of organic foods, whereas no significant changes were observed in inorganic treated groups. Moreover, MGEs, including IS613, Tp614 and tnpA_03 were found to play an important role in regulating ARG profiles in the gut microbiome following consumption of organic foods. Importantly, feeding organic food increased the relative abundance of the potentially antibiotic-resistant pathogens, Bacteroides and Streptococcus. Our results confirm that there is an increasing risk of ARGs and ARB in the gut microbiome, which highlights the importance of organic food industries taking into account the potential accumulation and transmission of ARGs as a risk factor.

Geographic characteristics and environmental variables determine the diversities and assembly of the algal communities in interconnected river-lake system.

Algal blooms in lakes are a major hazard worldwide. Although various geographical and environmental patterns affect algal communities during river-lake transit, a thorough understanding of what patterns shape the algal communities is still rarely researched, particularly in complex interconnected river-lake systems. In this study, focusing on the most typical interconnected river-lake system in China, the Dongting Lake, we collected paired water and sediment samples in summer, when algal biomass and growth rate are at high levels. Based on 23S rRNA gene sequencing, we investigated the heterogeneity and the differences in assembly mechanisms of planktonic and benthic algae in Dongting Lake. Planktonic algae contained more Cyanobacteria and Cryptophyta, while sediment harbored higher proportions of Bacillariophyta and Chlorophyta. For planktonic algae, stochastic dispersal dominated the assembly of the communities. Upstream rivers and confluences were important sources of planktonic algae in lakes. Meanwhile, for benthic algae, deterministic environmental filtering shaped the communities, and the proportion of benthic algae exploded with increasing N:P ratio and Cu concentration until reaching thresholds of 1.5 and 0.013 g/kg respectively, and then started falling, showing non-linear responses. This study revealed the variability of different aspects of algal communities in different habitats, traced the main sources of planktonic algae, and identified the thresholds for benthic algal shifts in response to environmental filters. Hence, upstream and downstream monitoring as well as thresholds of environmental factors should be considered in further aquatic ecological monitoring or regulatory programs of harmful algal blooms in these complex systems.

Engineering artificial microbial consortia based on division of labor promoted simultaneous removal of Cr(VI)-atrazine combined pollution.

Combined pollution caused by organic pollutants and heavy metals is common in polluted sites and wastewater. Engineering artificial microbial consortia offers a promising approach to address this complex issue. However, the mutualistic interactions and the critical function of specific microbe within microbial consortia remain unclear. In this study, based on division of labor, we respectively co-cultured two Cr(VI)-reducing strains, Paenarthrobacter nitroguajacolicus C1 and Pseudomonas putida C2, with an atrazine-degrading strain, Paenarthrobacter ureafaciens AT. After 5 days, up to 95 % Cr(VI) and 100 % atrazine were removed from the cocultures. Strain AT degraded nearly all atrazine and contributed only to a fraction of Cr(VI) reduction, whereas C1 promoted 41 % Cr(VI) transformation to Cr(III) fixed in cells, and C2 promoted 91 % Cr(VI) transformation to soluble Cr(III). Metabolic analyses of the cocultures and monocultures demonstrated that AT provided C1 with isopropylamine by passive diffusion and C2 with other effective nitrogen resources by cell-cell surface contact to promote their growth. Soil experiments also showed that treatments with AT and C2 achieved the highest Cr(VI) reduction and no atrazine residue. Our results indicate that engineering artificial microbial consortia based on division of labor and metabolic interactions is effective in promoting highly efficient bioremediation of combined pollution.

Phylogenetic distance affects the artificial microbial consortia's effectiveness and colonization during the bioremediation of polluted soil with Cr(VI) and atrazine.

Soils co-contaminated with heavy metals and organic pollutants are common and threaten the natural environment and human health. Although artificial microbial consortia have advantages over single strains, the mechanism affecting their effectiveness and colonization in polluted soils still requires determination. Here, we constructed two kinds of artificial microbial consortia from the same or different phylogenetic groups and inoculated them into soil co-contaminated with Cr(VI) and atrazine to study the effects of phylogenetic distance on consortia effectiveness and colonization. The residual concentrations of pollutants demonstrated that the artificial microbial consortium from different phylogenetic groups achieved the highest removal rates of Cr(VI) and atrazine. The removal rate of 400 mg/kg atrazine was 100%, while that of 40 mg/kg Cr(VI) was 57.7%. High-throughput sequence analysis showed that the soil bacterial negative correlations, core genera, and potential metabolic interactions differed among treatments. Furthermore, artificial microbial consortia from different phylogenetic groups had better colonization and a more significant effect on the abundance of native core bacteria than consortia from the same phylogenetic group. Our study highlights the importance of phylogenetic distance on consortium effectiveness and colonization and offers insight into the bioremediation of combined pollutants.

Protistan consumers and phototrophs are more sensitive than bacteria and fungi to pyrene exposure in soil.

The levels of organic pollutants, in particular polycyclic aromatic hydrocarbons (PAHs), are increasing worldwide, yet we lack clarity on how these pollutants affect microbial communities of different trophic levels, including protists, fungi, and bacteria. Herein, we conducted soil microcosm incubation experiments to investigate the effects of pyrene, a typical PAH, on microbial communities along concentration gradients from 0 to 500 mg kg-1 soil. Protistan communities were more sensitive to pollutants than fungal and bacterial communities, and protistan consumers and phototrophs were the dominant trophic functional groups. In addition, by assessing changes in the diversity and structure of the soil microbiome and ecological networks, we found that the microbial communities, including the protistan community and the two trophic communities composed of protists and their prey, were destabilized with increasing stress and pyrene concentrations. We identified links and complicated relationships between phototrophs, bacteria, and consumers in food webs, which explain the importance of protists in stabilizing the microbial community. Collectively, our work provides novel evidence that protists are considerably sensitive to pollution stress, and caution should be exercised in future evaluations of the protistan and multitrophic communities in polluted soil ecosystems.

Nitrogen Removal Characteristics and Constraints of an Alphaproteobacteria with Potential for High Nitrogen Content Heterotrophic Nitrification-Aerobic Denitrification.

The discovery of heterotrophic nitrification-aerobic denitrification (HN-AD) microorganisms has opened a new window for wastewater treatment. The underlying mechanism of HN-AD, however, is not fully understood because of the phylogenetic diversity of HN-AD microbes. The isolation and characterization of new HN-AD microorganisms are encouraging for furthering the understanding of this process. In this study, we found an Alphaproteobacteria isolate W30 from a historically polluted river in China through an HN-AD microbes screening process, which we identified as Pannonibacter sp. A potential HN-AD pathway for W30 was proposed based on N conversion analyses and the successful amplification of the entire denitrification gene series. The isolate exhibited high efficiency of aerobic inorganic nitrogen transformation, which accounted for 97.11% of NH4+-N, 100% of NO3--N, and 99.98% of NO2--N removal with a maximum linear rate of 10.21 mg/L/h, 10.46 mg/L/h, and 10.77 mg/L/h, respectively. Assimilation rather than denitrification was the main mechanism for the environmental nitrogen depletion mediated by W30. The effect of environmental constraints on aerobic NO3--N removal were characterized, following a membrane bioreactor effluent test under an oxic condition. Compared to known Alphaproteobacterial HN-AD microbes, we showed that Pannonibacter sp. W30 could deplete nitrogen with no NO2--N or NO3--N accumulation in the HN-AD process. Therefore, the application of Pannonibacter sp. W30 has the potential for developing a felicitous HN-AD technology to treat N-laden wastewater at the full-scale level.

The Immediate Hotspot of Microbial Nitrogen Cycling in an Oil-Seed Rape (Brassica campestris L.) Soil System Is the Bulk Soil Rather Than the Rhizosphere after Biofertilization.

Biofertilizers are substances that promote plant growth through the efficacy of living microorganisms. The functional microbes comprising biofertilizers are effective mediators in plant-soil systems in the regulation of nitrogen cycling, especially in nitrification repression. However, the deterministic or stochastic distribution of the functional hotspot where microbes are active immediately after biofertilization is rarely investigated. Here, pot experiments with oil-seed rape (Brassica campestris L.) were conducted with various chemical and biological fertilizers in order to reveal the distribution of the hotspot after each fertilization. A stimulated dynamic of the nitrogen cycling-related genes in the bulk soil inferred that the bulk soil was likely to be the hotspot where the inoculated bacterial fertilizers dominated the nitrogen cycle. Furthermore, a network analysis showed that bulk soil microbial communities were more cooperative than those in the rhizosphere after biofertilization, suggesting that the microbiome of the bulk soils were more efficient for nutrient cycling. In addition, the relatively abundant ammonia-oxidizing bacteria and archaea present in the networks of bulk soil microbial communities further indicated that the bulk soil was the plausible hotspot after the application of the biofertilizers. Therefore, our research provides a new insight into the explicit practice of plant fertilization and agricultural management, which may improve the implementational efficiency of biofertilization.