Long-term nitrogen fertilization-induced enhancements of acid hydrolyzable nitrogen are mainly regulated by the most vital microbial taxa of keystone species and enzyme activities.

It is well known that nitrogen (N) fertilizer input is required to improve crop productivity, but we lack a comprehensive understanding of how elevated N input changes the formation of soil acid hydrolyzable nitrogen (AHN) by adjusting the most vital microbial taxa of keystone species of microbial communities and enzyme activities. A 15-year field experiment comprising four levels of inorganic N fertilization was conducted to identify the most important bacterial and fungal taxa of the keystone species derived from cooccurrence networks as well as the vital enzyme activities at the bell mouth and maturity stages. Long-term N fertilization significantly increased the levels of AHN along with its four fractions, including amino acid N (AAN), ammonium N (AN), amino sugar N (ASN), and hydrolysable unidentified N (HUN), by 30.1-118.6 %, regardless of growth stage. Some most vital microbial taxa of keystone species and enzyme activities, which changed in response to N fertilization, mainly regulated each ANH fraction, that is, AHN and AN were mainly controlled by the enrichment of Nocardioides and ß-1,4-N-acetyl-glucosaminidase (NAG), as well as by the reduction of Anaerolinea and urease (UR), AAN was determined by the enrichment of Hannaella and depletion of Penicillium, ASN was regulated by the enrichment of Hannaella and Arthrobacter, and HUN was influenced by the reduction of Penicillium and enrichment of Nitrosospira. These microbial genera have been found to be involved in dissimilatory nitrate reduction to ammonium (DNRA) and nitrification/denitrification processes and the two enzyme activities involved in organic N degradation and N-releasing processes, suggesting that the formation of AHN fractions was closely associated with specific functional microbial taxa and enzyme activities induced by N fertilization. Our results provide new insights into the associations among increased N input, altered formation of soil organic N, and shifts in microbial communities and enzyme activities.

Biochar-induced variations in crop yield are closely associated with the abundance and diversity of keystone species.

Biochar application has widely been used to improve crop yield, but its effectiveness is uncertain. Soil microbial communities may play critical roles, but we lack experimental evidences on the relationships between these communities and crop yield following biochar application. Here, we used cooccurrence networks to demonstrate the importance of ecological clusters (cooccurring taxa of soil microbes including bacteria and fungi) and to identify specific keystone species that were closely connected with the variations in crop yield in a pot experiment. The experiment included two soils (i.e., red soil and yellow-cinnamon soil) for wheat growth, with each soil receiving three biochar application rate. The grain yield in the red soil significantly increased while that in the yellow-cinnamon soil significantly decreased with the biochar application rate. Generally, the grain yield from the two soils showed close correlations with the relative abundance as well as with the diversity of keystone species within major clusters rather than with the soil properties and enzyme activities. This contrasting effectiveness was mainly associated with the enrichment of beneficial and suppression of detrimental keystone species in the red soil and the suppression of beneficial and enrichment of detrimental keystone species in the yellow-cinnamon soil. These species together mainly accounted for the variation in the relative abundance of keystone species within major clusters of each soil, suggesting their potential to affect crop yield following biochar application.

Salt tolerance-based niche differentiation of soil ammonia oxidizers.

Ammonia oxidizers are key players in the global nitrogen cycle, yet little is known about their ecological performances and adaptation strategies for growth in saline terrestrial ecosystems. This study combined 13C-DNA stable-isotope probing (SIP) microcosms with amplicon and shotgun sequencing to reveal the composition and genomic adaptations of active ammonia oxidizers in a saline-sodic (solonetz) soil with high salinity and pH (20.9 cmolc exchangeable Na+ kg-1 soil and pH 9.64). Both ammonia-oxidizing archaea (AOA) and bacteria (AOB) exhibited strong nitrification activities, although AOB performed most of the ammonia oxidation observed in the solonetz soil and in the farmland soil converted from solonetz soil. Members of the Nitrosococcus, which are more often associated with aquatic habitats, were identified as the dominant ammonia oxidizers in the solonetz soil with the first direct labeling evidence, while members of the Nitrosospira were the dominant ammonia oxidizers in the farmland soil, which had much lower salinity and pH. Metagenomic analysis of "Candidatus Nitrosococcus sp. Sol14", a new species within the Nitrosococcus lineage, revealed multiple genomic adaptations predicted to facilitate osmotic and pH homeostasis in this extreme habitat, including direct Na+ extrusion/H+ import and the ability to increase intracellular osmotic pressure by accumulating compatible solutes. Comparative genomic analysis revealed that variation in salt-tolerance mechanisms was the primary driver for the niche differentiation of ammonia oxidizers in saline-sodic soils. These results demonstrate how ammonia oxidizers can adapt to saline-sodic soil with excessive Na+ content and provide new insights on the nitrogen cycle in extreme terrestrial ecosystems.

Demonstration of Chemical Distinction among Soil Humic Fractions Using Quantitative Solid-State 13C NMR.

Humic substances (HS) are vital to soil fertility and carbon sequestration. Using multiple cross-polarization/magic-angle spinning (multiCP/MAS) NMR combined with dipolar dephasing, we quantitatively characterized humic fractions, i.e., fulvic acid (FA), humic acid (HA), and humin (HM), isolated from two representative soils (upland and paddy soils) in China under six long-term (>20 years) fertilizer treatments. Results indicate that each humic fraction showed chemical distinction between the upland and paddy soils, especially with much greater aromaticity of upland HMs than of paddy HMs. Fertilizer treatment exerted greater influence on chemical natures of upland HS than of paddy HS, although the effect was less than that of soil type. Organic manure application especially decreased the percentages of aromatic C in the upland HAs and HMs compared with the control. We concluded that humic fractions responded in chemical nature to environmental conditions, i.e., soil type/cropping system/soil aeration and fertilizer treatments.

Divergent responses of bacterial activity, structure, and co-occurrence patterns to long-term unbalanced fertilization without nitrogen, phosphorus, or potassium in a cultivated vertisol.

Unbalanced fertilization lacking nitrogen (N), phosphorus (P), or potassium (K) is a worldwide phenomenon; however, whether they affect bacterial community composition and intraspecific interactions in a similar pattern and how they affect bacterial activity are not systematically compared. Soils under different kinds of unbalanced fertilization in a 21-year field experiment were collected to investigate the variation in dehydrogenase activity (DHA), bacterial community diversity, structure, composition, and possible interactions. Compared to the balanced fertilization of NPK, the DHA from unbalanced fertilization of NP, PK, and NK was 8.70, 11.59, and 14.17% lower, respectively, and from the unfertilized treatment (Nil) was 13.41% lower; however, the Shannon index from NP, PK, and Nil was 4.48-7.21% higher and from NK was 3.95% lower. Based on principal coordinate analyses (PCoA), bacterial community structure was separated by N application or not along PCo1 and was further separated by P application or not along PCo2, indicating a more influence by N deficiency. Moreover, the structure was mainly determined by soil pH, soil organic carbon (SOC), and total phosphorus (TP). The network complexity using co-occurrence analysis followed the order NP > NPK > PK > NK > Nil, indicating a more influence by P deficiency on intraspecific interactions. Structural equation modeling (SEM) revealed that the reduced DHA in NP was mainly regulated by the decreased SOC and increased Shannon index, in PK by the decreased SOC and increased Shannon index and pH, and in NK by the decreased SOC and TP and increased PCo2. The significantly lower abundance of Bacteroidetes and Chitinophagaceae in NK may also contribute to the reduced DHA. Our results imply that N deficiency had the greatest impact on bacterial community structure and composition, P deficiency had the greatest impact on network construction and bacterial activity, and K deficiency has minimal effect. Our results also suggest that main factors regulating the variation in soil functions may vary among different nutrient deficiencies.

Potential Root Foraging Strategy of Wheat (Triticum aestivum L.) for Potassium Heterogeneity.

Potassium (K) distribution is horizontally heterogeneous under the conservation agriculture approach of no-till with strip fertilization. The root foraging strategy of wheat for K heterogeneity is poorly understood. In this study, WinRHIZO, microarray, Non-invasive Micro-test Technology (NMT) and a split-root system were performed to investigate root morphology, gene expression profiling and fluxes of K+ and O2 under K heterogeneity and homogeneity conditions. The split-root system was performed as follows: C. LK (both compartments had low K), C. NK (both compartments had normal K), Sp. LK (one compartment had low K) and Sp. NK (the other compartment had normal K). The ratio of total root length and root tips in Sp. NK was significantly higher than that in C. NK, while no significant differences were found between Sp. LK and C. LK. Differential expression genes in C. LK vs. C. NK had opposite responses in Sp. LK vs. C. LK and similar responses in Sp. NK vs. C. NK. Low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding, glutathione transferase and cellular respiration genes were found to be up-regulated in Sp. NK. However, methyltransferase activity, protein amino acid phosphorylation, potassium ion transport, and protein kinase activity genes were found to be down-regulated in Sp. LK. The up-regulated gene with function in respiration tended to increase K+ uptake through improving O2 influx on the root surface in Sp. NK, while the down-regulated genes with functions of K+ and O2 transport tended to reduce K+ uptake on the root surface in Sp. LK. To summarize, wheat roots tended to perform active-foraging strategies in Sp. NK and dormant-foraging strategies in Sp. LK through the following patterns: (1) root development in Sp. NK but not in Sp. LK; (2) low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding and respiration, were up-regulated in Sp. NK but not in Sp. LK; and (3) root K+ and O2 influxes increased in Sp. NK but not in Sp. LK. Our findings may better explain the optimal root foraging strategy for wheat grown with heterogeneous K distribution in the root zone.

Effects of residue incorporation and plant growth on soil labile organic carbon and microbial function and community composition under two soil moisture levels.

This study investigates the effects of residue incorporation coupled with plant growth and soil moisture level on wheat biomasses, soil nutrients, labile organic carbon (LOC), microbial metabolic profiles, and community composition. Four management practices were used in a 180-day pot experiment: (1) control (CON), (2) maize (Zea mays L.) residue incorporation without plants (MR), (3) wheat (Triticum aestivum L.) plants without maize residue (WP), and (4) maize residue incorporation with wheat plants (MRWPs). Each management practice included soil moisture at both 40 and 80% of field capacity. At wheat harvest, soil nutrient contents in the WP and MRWP treatments were significantly lower than in the CON and MR treatments. In comparison with the CON treatment, MR, WP, and MRWP treatments resulted in 35, 23, and 67% increases in dissolved organic carbon content; 17, 12, and 34% increases in hot-water extractable organic carbon content; and 78, 50, and 150% increases in microbial biomass carbon content. Furthermore, microbial utilizations of carboxylic acids and polymer carbon sources in the MR, WP, and MRWP treatments were 261 and 88%, 239 and 105%, and 300 and 126% higher than in the CON treatment. The MR and CON treatments had similar phospholipid fatty acid (PLFA) content but the WP and MRWP treatments had significantly increased gram-negative content and changes to community composition compared with the CON and MR treatments. The wheat biomass, LOC, and PLFA contents significantly increased with greater soil moisture. Overall, these results suggest an additive effect of residue incorporation and plant growth on LOC contents, primarily due to the changes in microbial utilization of carbon sources and community composition.

Altered humin compositions under organic and inorganic fertilization on an intensively cultivated sandy loam soil.

Humin is the most recalcitrant fraction of soil organic matter (SOM). However, little is known about quantitative structural information on humin and the roles of soil mircoorganisms involved in the humin formation. We applied advanced solid-state 13C nuclear magnetic resonance (NMR) spectroscopy to provide deep insights into humin structural changes in response to long-term balanced fertilization on a Calcaric Fluvisol in the North China plain. The relationships between humin structure and microbiological properties such as microbial biomass, microbial quotient (qmic) and metabolic quotient (qCO2) were also studied. The humins had a considerable (35-44%) proportion of aromatic C being nonprotonated and the vast majority of O-alkyl and anomeric C being protonated. Alkyl (24-27% of all C), aromatic C (17-28%) and O-alkyl (13-20%) predominated in humins. Long-term fertilization promoted the aliphatic nature of humins, causing increases in O-alkyl, anomeric and NCH functional groups and decreases in aromatic C and aromatic CO groups. All these changes were more prominent for treatments of organic fertilizer (OF) and combined mineral NPK fertilizer with OF (NPKOF) relative to the Control and NPK treatments. Fertilization also decreased the alkyl/O-alkyl ratio, aromaticity and hydrophobic characteristics of humins, suggesting a more decomposed and humified state of humin in the Control soil. Moreover, the soil microbiological properties had strong correlations with functional groups of humins. Particularly, microbial biomass C was a relatively sensitive indicator, having positive correlations with oxygen-containing functional groups, i.e., COO/NCO and protonated O-alkyl C, and negative correlations with nonprotonated aromatic C. The qmic and qCO2 were also significantly positively correlated with NCH and aromatic CO, respectively. Our results deepen our understanding of how long-term fertilization impacts the structure of humin, and highlight a linkage between microbiological properties and recalcitrant fraction of SOM besides labile fraction.

Chemical nature of humic substances in two typical Chinese soils (upland vs paddy soil): A comparative advanced solid state NMR study.

Knowledge of the structural features of humic substances (HSs) is essential for elucidating the mechanisms of humification in different soil environments and realizing their profound roles in environmental issues. The aim of this work was to investigate the chemical structures of fulvic acid (FA), humic acid (HA) and humin (HM) fractions isolated from an upland soil (Fluvisol) and a paddy soil (Anthrosol) typical in China using advanced solid-state 13C nuclear magnetic resonance (NMR) techniques. The results revealed that there were great structural differences of HSs between the two soils. The two FAs showed distinct quantitative differences in aliphatics with more polysaccharides in the FA from the upland soil than from the paddy soil. The HM from the upland soil differed from the paddy soil HM in having more proteins/peptides (23% vs 20%), total aromatics (21% vs 12%) as well as fewer lipids (24% vs 35%) and polysaccharides (27% vs 31%). The HM fractions represented the most different components of organic matter between the two soils. The degree of difference between the two HAs fell in between that of FAs and HM fractions. In particular, the HA from the upland soil had relatively greater degree of aromaticity. Our study indicated that the upland soil exhibited a higher degree of humification compared with the paddy soil. Among the three humic fractions, the FAs featured COO/N-CO groups, and the HAs were more enriched in protonated aromatic C for both soils. In contrast, the two HM fractions contained more O-alkyl C and fewer aromatics than did the other humic fractions, being closer to the original organic materials in soils. We speculate that the evolutionary route of HSs is likely to be the transformation of original organic materials into HM, followed by increased degradation, further oxidization and conversion into HA, and then into FA.

Crop residue management and fertilization effects on soil organic matter and associated biological properties.

Returning crop residue may result in nutrient reduction in soil in the first few years. A two-year field experiment was conducted to assess whether this negative effect is alleviated by improved crop residue management (CRM). Nine treatments (3 CRM and 3 N fertilizer rates) were used. The CRM treatments were (1) R0: 100 % of the N using mineral fertilizer with no crop residues return; (2) R: crop residue plus mineral fertilizer as for the R0; and (3) Rc: crop residue plus 83 % of the N using mineral and 17 % manure fertilizer. Each CRM received N fertilizer rates at 270, 360, and 450 kg N ha(-1) year(-1). At the end of the experiment, soil NO3-N was reduced by 33 % from the R relative to the R0 treatment, while the Rc treatment resulted in a 21 to 44 % increase in occluded particulate organic C and N, and 80 °C extracted dissolved organic N, 19 to 32 % increase in microbial biomass C and protease activity, and higher monounsaturated phospholipid fatty acid (PLFA):saturated PLFA ratio from stimulating growth of indigenous bacteria when compared with the R treatment. Principal component analysis showed that the Biolog and PLFA profiles in the three CRM treatments were different from each other. Overall, these properties were not influenced by the used N fertilizer rates. Our results indicated that application of 17 % of the total N using manure in a field with crop residues return was effective for improving potential plant N availability and labile soil organic matter, primarily due to a shift in the dominant microorganisms.

Effects of changes in straw chemical properties and alkaline soils on bacterial communities engaged in straw decomposition at different temperatures.

Differences in the composition of a bacterial community engaged in decomposing wheat straw in a fluvo-aquic soil at 15 °C, 25 °C, and 35 °C were identified using barcode pyrosequencing. Functional carbon groups in the decomposing wheat straw were evaluated by (13)C-NMR (nuclear magnetic resonance). Actinobacteria and Firmicutes were more abundant, whereas Alphaproteobacteria and Bacteroidetes were less abundant, at higher temperatures during the later stages of decomposition. Differences in the chemical properties of straw accounted for 19.3% of the variation in the community composition, whereas soil properties accounted for more (24.0%) and temperature, for less (7.4%). Carbon content of the soil microbial biomass and nitrogen content of straw were significantly correlated with the abundance of Alphaproteobacteria, Actinobacteria, and Bacteroidetes. The chemical properties of straw, especially the NCH/OCH3, alkyl O-C-O, and O-alkyl functional groups, exercised a significant effect on the composition of the bacterial community at different temperatures during decomposition-results that extend our understanding of bacterial communities associated with the decomposition of straw in agro-ecosystems and of the effects of temperature and chemical properties of the decomposing straw and soil on such communities.

Mass loss and chemical structures of wheat and maize straws in response to ultraviolet-B radiation and soil contact.

The role of photodegradation, an abiotic process, has been largely overlooked during straw decomposition in mesic ecosystems. We investigated the mass loss and chemical structures of straw decomposition in response to elevated UV-B radiation with or without soil contact over a 12-month litterbag experiment. Wheat and maize straw samples with and without soil contact were exposed to three radiation levels: a no-sunlight control, ambient solar UV-B, and artificially elevated UV-B radiation. A block control with soil contact was not included. Compared with the no-sunlight control, UV-B radiation increased the mass loss by 14-19% and the ambient radiation by 9-16% for wheat and maize straws without soil contact after 12 months. Elevated UV-B exposure decreased the decomposition rates of both wheat and maize straws when in contact with soil. Light exposure resulted in decreased O-alkyl carbons and increased alkyl carbons for both the wheat and maize straws compared with no-sunlight control. The difference in soil contact may influence the contribution of photodegradation to the overall straw decomposition process. These results indicate that we must take into account the effects of photodegradation when explaining the mechanisms of straw decomposition in mesic ecosystems.

Comparative analysis of potassium deficiency-responsive transcriptomes in low potassium susceptible and tolerant wheat (Triticum aestivum L.).

Potassium (K(+)) deficiency as a common abiotic stress can inhibit the growth of plants and thus reduce the agricultural yields. Nevertheless, scarcely any development has been promoted in wheat transcriptional changes under K(+) deficiency. Here we investigated root transcriptional changes in two wheat genotypes, namely, low-K(+) tolerant "Tongzhou916" and low-K(+) susceptible "Shiluan02-1". There were totally 2713 and 2485 probe sets displayed expression changes more than 1.5-fold in Tongzhou916 and Shiluan02-1, respectively. Low-K(+) responsive genes mainly belonged to the categories as follows: metabolic process, cation binding, transferase activity, ion transporters and so forth. We made a comparison of gene expression differences between the two wheat genotypes. There were 1321 and 1177 up-regulated genes in Tongzhou916 and Shiluan02-1, respectively. This result indicated that more genes took part in acclimating to low-K(+) stress in Tongzhou916. In addition, there were more genes associated with jasmonic acid, defense response and potassium transporter up-regulated in Tongzhou916. Moreover, totally 19 genes encoding vacuolar H(+)-pyrophosphatase, ethylene-related, auxin response, anatomical structure development and nutrient reservoir were uniquely up-regulated in Tongzhou916. For their important role in root architecture, K(+) uptake and nutrient storage, unique genes above may make a great contribution to the strong low-K(+) tolerance in Tongzhou916.

Differences in chemical composition of soil organic carbon resulting from long-term fertilization strategies.

Chemical composition of soil organic carbon (SOC) is central to soil fertility. We hypothesize that change in SOC content resulting from various long-term fertilization strategies accompanies the shift in SOC chemical structure. This study examined the effect of fertilization strategies along with the time of fertilizer application on the SOC composition by 13C nuclear magnetic resonance (NMR) spectroscopy. The soils (Aquic Inceptisol) subjected to seven fertilizer treatments were collected in 1989, 1999 and 2009, representing 0, 10 and 20 years of fertilization, respectively. The seven fertilizer treatments were (1-3) balanced fertilization with application of nitrogen (N), phosphorus (P) and potassium (K) including organic compost (OM), half organic compost plus half chemical fertilizer (1/2OM), and pure chemical NPK fertilizer (NPK); (4-6) unbalanced chemical fertilization without application of one of the major elements including NP fertilizer (NP), PK fertilizer (PK), and NK fertilizer (NK); and (7) an unamended control (CK). The SOC content in the balanced fertilization treatments were 2.3-52.6% and 9.4-64.6% higher than in the unbalanced fertilization/CK treatments in 1999 and 2009, respectively, indicating significant differences in SOC content with time of fertilizer application between the two treatment groups. There was a significantly greater proportion of O-alkyl C and a lower proportion of aromatic C in the balanced fertilization than in unbalanced fertilization/CK treatments in 1999, but not in 2009, because their proportions in the former treatments approached the latter in 2009. Principal component analysis further showed that the C functional groups from various fertilization strategies tended to become compositionally similar with time. The results suggest that a shift in SOC chemical composition may be firstly dominated by fertilization strategies, followed by fertilization duration.

The influence of long-term fertilization on cadmium (Cd) accumulation in soil and its uptake by crops.

Continuous application of organic and inorganic fertilizers can affect soil and food quality with respect to heavy metal concentrations. The risk of cadmium (Cd) contamination in a long-term (over 20 years) experimental field in North China with an annual crop rotation of winter wheat and summer maize was investigated. The long-term experiment had a complete randomized block design with seven fertilizer treatments and four replications. The seven fertilizer treatments were (1) organic compost (OM), (2) half organic compost plus half chemical fertilizer (OM + NPK), (3) NPK fertilizer (NPK), (4-6) chemical fertilizers without one of the major nutrients (NP, PK, and NK), and (7) an unamended control (CK). Soil samples from 0 to 20 cm were collected in 1989, 1999, and 2009 to characterize Cd and other soil properties. During the past 20 years, various extents of Cd accumulation were observed in the soil, and the accumulation was mainly affected by atmospheric dry and wet deposition and fertilization. In 2009, the average Cd concentration in the soil was 148 ± 15 µg kg(-1) and decreased in the order of NPK ≈ OM + NKP ≈ PK > NP ≈ NK > OM ≈ CK. Sequential extraction of Cd showed that the acid-soluble fraction (F1, 32 ± 7 %) and the residual fraction (F4, 31 ± 5 %) were the dominant fractions of Cd in the soil, followed by the reducible fraction (F2, 22 ± 5 %) and oxidizable fraction (F3, 15 ± 6 %). The acid-soluble Cd fraction in the soil and Cd accumulation in the crops increased with soil plant available K. Fraction F3 was increased by soil organic C (SOC) and crop yields, but SOC reduced the uptake of soil Cd by crops. The long-term P fertilization resulted in more Cd buildup in the soil than other treatments, but the uptake of Cd by crops was inhibited by the precipitation of Cd with phosphate in the soil. Although soil Cd was slightly increased over the 20 years of intensive crop production, both soil and grain/kernel Cd concentrations were still below the national standards for environmental and food safety.

Function of bacterial cells and their exuded extracellular polymeric substances (EPS) in virus removal by red soils.

The potential influence of autochthonous microorganisms on virus fate in soil is usually determined through extreme conditions of sterilization vs. nonsterilization; however, the relative importance of microbial cells and their exudates remains unclear. In this study, bacterial cells (cell) were harvested, and their exuded extracellular polymeric substances (EPS) were extracted from three strains of bacteria, namely, Gram-negative bacteria Pseudomonas putida and Pseudomonas aeruginosa as well as Gram-positive bacterium Bacillus subtilis. This study aimed to evaluate virus removal in solutions in the presence of cell, EPS, and their combination (cell/EPS), as well as to investigate how their presence affects virus removal efficiencies by four red soils based on batch experiments. Results showed that virus removal percentage in solutions ranged from 11 to 23 in the presence of cells only and from 12 to 15 in the presence of EPS only. The removal percentage in the combined cell/EPS treatment can be estimated by summing the results achieved by the cell and EPS treatments, separately. Meanwhile, cell presence had a negligible effect on virus removal by red soils. EPS and combined cell/EPS significantly reduced virus removal by 20 to 69% and 16 to 50%, respectively, which indicated that EPS served a dominant function in reducing virus removal. This study clearly demonstrated that the prediction of virus removal by red soils must consider the effect of bacteria, especially those producing large quantities of EPS, which can be responsible for the underestimation of viral load in certain studies.

Presence of bacteria in aqueous solution influences virus adsorption on nanoparticles.

Virus contamination in wastewater is usually accompanied by the existence of various bacteria. Nanoparticles (NPs) have been shown to efficiently remove virus. In this study, bacterial cells, supernatants, and cultures were harvested separately from three strains at the culture ages of 6 and 24 h, corresponding to the log and stationary phases, respectively. The aim is to investigate how their presence affects virus adsorption on the three Fe and Al oxide NPs (α-Fe2O3, γ-Fe2O3-B, and Al2O3) and how these effects change with bacterial growth phase. Bacteriophage phiX174 was used as a virus model. Results showed that bacterial cells, supernatants, and cultures harvested at 6 h generally reduced virus adsorption by an average of 0.75±0.84, 7.7±9.0, and 10.3±8.6%, respectively, while those harvested at 24 h reduced virus adsorption by an average of 2.1±0.93, 21.5±6.6, and 24.6±6.9%, respectively. Among the NPs, α-Fe2O3 showed more sensitivity to bacteria than the other two, probably because of its relatively higher value of point of zero charge. It was found that cell-induced and supernatant-induced reductions were combined to achieve added results, in which the supernatants contributed much more than the cells, implying that the bacterial exudates might be more crucial in the reduced virus adsorption than the bacterial cells. These results strongly demonstrated that the bacteria-induced reduction in virus adsorption became more significant with culture age. It is suggested that studies conducted in the absence of bacteria may not accurately evaluate the potential of virus removal efficiency of the NPs in bacteria-containing environments.

[Virus adsorption onto nano-sized iron oxides as affected by different background solutions].

Batch adsorption studies were conducted to investigate virus adsorption onto four commercial nanoparticles of iron oxide as affected by different background solutions, using bacteriophage phiX174 as virus indicator. When artificial ground water was used, the 4 studied nanoparticles showed high virus adsorption capacity, among which alpha-Fe2O3 was the most effective, with the adsorption percent reaching 100% at low initial virus concentration (i.e. 1E + 03 PFU x mL(-1)). Virus adsorption results were described using the Langmuir and Freundlich adsorption isotherms. The estimated adsorption parameters indicated the presence of multilayer adsorption and favorable adsorption. The adsorption percentage by the studied nanoparticles increased with decreasing virus initial concentration. Our results further showed that higher ionic strength of the background solution reduced the virus adsorption, indicating that electrostatic interaction likely dominated the virus adsorption. The presence of anions in the background solution reduced the virus adsorption, probably because of the competitive adsorption between the viruses and anions for sorption sites available, among which HPO4(2-) showed more reduced than HCO3(-). On the other hand, the presence of multivalent cations was favorable for virus adsorption, with bivalent cations (e.g. Ca2+ and Mg2+) showing more favorable than monovalent cations (e.g. Na+ and K+). Results of this study suggest that nanoparticles of iron oxide may be potentially useful for virus removal from infecting water, while other anions or cations in the water should be considered.

Removal of bacteriophages MS2 and phiX174 from aqueous solutions using a red soil.

Adsorption and desorption of viruses onto and from an adsorbent may have a dominant role in evaluating removal efficiency of a material. This study evaluated the effectiveness of a red soil from south part of China to remove two viruses, MS2 and phiX174, by adsorption from dilute aqueous solutions using a set of equilibrium and kinetic batch experiments. The effect of presence/absence of autochthonous microorganisms was also investigated. The results showed that when the autochthonous microorganisms were present, the red soil adsorbed more than 99.95% of MS2 and 98.23% of phiX174, in which most of them were inactivated and/or irreversibly adsorbed. Sterilization led to an increase in MS2 adsorption, while decreased the adsorption of phiX174, indicating that sterilization-induced virus adsorption is virus type dependent. Fewer viruses could be desorbed from the sterilized soil as compared to the nonsterilized soil, probably because sterilization led to an increase in the strength of adsorption force between the soil and viruses. Though the overall virus removal efficiency by the red soil was less than the USPEA required value of 99.99%, we suggest the potential of the red soil as a starting material in removing water heavily polluted with viruses.

[Numerical simulation of isothermal equilibrium adsorption of virus onto typical soils in China].

An isothermal batch experiment was conducted in the laboratory to compare adsorption of bacteriophages MS2 and phiX174 onto 6 different soils (red loam soil, red clay soil, wushan soil, huangni soil, sandy fluvo-aquic soil and loamy fluvo-aquic soil) in China. Soils with sterilized or non-sterilized treatment were used. Relative coefficients of each numerical simulation of the isotherms using three models were evaluated. The results show that the properties of the soil and virus, and presence/absence of the soil autochthonous microorganisms have pronounced effect on the virus adsorption. Both MS2 and phiX174 are almost completely adsorbed by the red clay soil, but minimal adsorption is observed in the two fluvo-aquic soils. Adsorption of phiX174 to all the non-sterilized soils is generally much greater than that of MS2, while sterilization leads to opposite results. Freundlich and Langmuir isotherms are found to have better coefficients to simulate the apparent steady-state virus concentrations. Freundlich isotherm is capable of demonstrating the effect of virus concentration on adsorption behavior. Langmuir isotherms can be used to compare relative adsorption among treatments, while the present study suggests that maximum adsorption can not be calculated when using the Langmuir isotherms.