Combination of intercropping maize and soybean with root exudate additions reduces metal mobility in soil-plant system under wastewater irrigation.

The effects of root exudates and irrigation with treated wastewater on heavy metal mobility and soil bacterial composition under intercropping remain poorly understood. We conducted a pot experiment with maize and soybean grown in monocultures or intercultures, irrigated with either groundwater or treated wastewater. In addition, the pre-collected root exudates from hydroponic culture with mono- or inter-cropped maize and soybean were applied to the soil at four levels (0 %, 16 %, 32 % and 64 %). The results showed that application of root exudates increased plant growth and soil nutrient content. The analysis of "Technique for Order of Preference by Similarity to Ideal Solution" for higher plant biomass and lower soil Cd and Pb concentrations indicated that the best performance of soybean under treated wastewater irrigation was recorded under intercropping applied with 64 % of exudates, with a performance score of 0.926 and 0.953 for Cd and Pb, respectively. The second-best performance of maize under treated wastewater irrigation was also observed under intercropping applied with 64 % of exudates. Root exudate application reduced heavy metals migration in the soil-plant system, with a greater impact in intercropping than in monocropping. In addition, certain soil microorganisms were also increased with root exudate application, regardless of irrigation water. This study suggests that appropriate application of root exudates could potentially improve plant growth and soil health, and reduce toxic heavy metal concentrations in soils and plants irrigated with treated wastewater.

Reduction effect of individual N, P, K fertilization on antibiotic resistance genes in reclaimed water irrigated soil.

The transfer of antibiotic resistance genes (ARGs) in soil under reclaimed water irrigation poses a potential environmental risk. Regulation of NPK fertilizer could influence the behavior of bacterial communities, mobile genetic elements (MGEs), and soil properties, which determine the fate of ARGs. To identify the key element in NPK fertilizer and realize efficient regulation, we explored the effect of individual N, P, K fertilization on ARG variation in tomato rhizosphere and bulk soils. Compared with an unfertilized treatment, N fertilization resulted in greater decreases in the abundance of ARGs (decreases of 24.06%-73.09%) than did either P fertilization (increases of up to 35.84%, decreases of up to 58.80%) or K fertilization (decreases of 13.47%-72.47%). The influence of different forms of N (CO(NH2)2, NaNO3, and NH4HCO3), P (Ca(H2PO4)2 and CaMgO4P+), and K (KCl and K2(SO4)) fertilizers was also investigated in this study, and showed the influence of NaNO3, CaMgO4P+, and K2(SO4) on reducing ARGs abundance was greater in different types of N, P, K fertilizers. Bacterial communities showed the strongest response to N fertilization. The reduced bacterial diversity and abundance of ARG-host and non-host organisms explained the decline of total ARG abundance in soil. In soils fertilized with either P or K, the effect of soil properties, especially total nitrogen and pH, on ARG variation was greater than that of bacterial community and MGEs. These results suggest that N regulation of in NPK fertilizer may be an effective way to reduce the risks of ARGs in soil associated with reclaimed water irrigation.

Variety features differentiate microbiota in the grape leaves.

The dependence of plant health and crop quality on the epiphytic microbial community has been extensively addressed, but little is known about plant-associated microbial communities under natural conditions. In this study, the bacterial and fungal communities on grape leaves were analyzed by 16S rRNA gene and internal transcribed spacer high-throughput sequencing, respectively. The results showed differences in the composition of the microbial communities on leaf samples of nine wine grape varieties. The most abundant bacterial genus was Pseudomonas, and the top three varieties with Pseudomonas were Zinfandel (22.6%), Syrah (21.6%), and Merlot (13.5%). The most abundant fungal genus was Alternaria, and the cultivar with the lowest abundance of Alternaria was Zinfandel (33.6%), indicating that these communities had different habitat preferences. The linear discriminant analysis effect size of all species showed that the bacteria Enterococcus, Massilia, and Kocuria were significantly enriched on the leaves of Merlot, Syrah, Cabernet Sauvignon, respectively; Pseudomonadales and Pantoea on Zinfandel; and Bacillus, Turicibacter, and Romboutsia on Pinot Noir. Similarly, the fungi Cladosporium, Phoma, and Sporormiella were significantly enriched on Zinfandel, Lon, and Gem, respectively. Both Bray-Curtis and unweighted UniFrac revealed that bacteria and fungi have a significant impact (P < 0.01), and the results further proved that variety is the most important factor affecting the microbial community. The findings indicate that some beneficial or harmful microorganisms existing on the wine grape leaves might affect the health of the grape plants and the wine-making process.

Formation of iron plaque on roots of Iris pseudacorus and its consequence for cadmium immobilization is impacted by zinc concentration.

The impact of iron plaque (IP) on bioavailability of heavy metals to plants has been well documented, but the role of zinc (Zn) in modulating the associated processes remains elusive. We took Iris pseudacorus used in wetland for remediating Cd-contaminated water as an example and systematically studied the combined influence of Cd and Zn concentration on formation of IP and its consequence for immobilization and plant uptake of Cd. The experiment was conducted in hydroponic culture and in each treatment, we measured the physiological traits, activity of antioxidant enzymes (SOD, POD, CAT), mass of the IP, as well as the Cd content in both plant tissues and IP. The results showed that increasing Cd concentration resulted in a steady reduction in IP while the impact of zinc on IP was complicated and appeared to be coupled with Cd. When the Cd concentration was low (0.5 mg L-1 measured as CdCl2 2·5H2O) increasing Zn concentration reduced IP, while when the Cd concentration was increased to 5 mg L-1 increasing zinc concentration led to an increase in IP mass first followed by a decline after Zn concentration exceeded 100 mg L-1 (measured as ZnSO4·7H2O). The change in IP as affected by Zn had a strong consequence for immobilization and plant uptake of Cd. When Cd concentration was low, the IP was comparatively abundant and hence adsorbed most Cd. In contrast, when Cd concentration was high, the IP reduced and the amount of Cd taken up by plant roots and translocated to shoots and leaves increased. Both Cd immobilization and its plant uptake were modulated by Zn concentration. At low Cd concentration the combined Cd immobilized and taken up by plant peaked when the Zn concentration was 50 mg L-1, while at high Cd concentration the combined Cd reached maxima when theZn concentration was 100 mg L-1. The activity of the antioxidant enzymes changed significantly with Zn rather than with Cd. Regardless of Cd concentration, the activity of all three antioxidant enzymes increased first with zinc concentration before declining when the Zn concentration exceeded approximately 100 mg L-1 in all treatments, comparable with the change in immobilization and plant uptake of Cd as the Zn concentration increased. SEM analysis did prove the formation and variation of IP on the root surface of Iris pseudacorus in different treatments. We also found that the plant developed a survival strategy by scarifying its leaves with high Cd content. The results presented in this paper has wide implications as it revealed that care needs to be taken in applying Zn to enhance Cd immobilization and its plant uptake as exceeding the optimal application rate might reduce remediating efficiency rather than increase it.

Investigating the bacterial community and amoebae population in rural domestic wastewater reclamation for irrigation.

Reclamation of domestic wastewater for agricultural irrigation is viewed as a sustainable option to create an alternative water source and address water scarcity. Free-living amoebae (FLA), which are amphizoic protozoa, are widely distributed in various environmental sources. The FLA could cause considerable environmental and health risks. However, little information is available on the risk of these protozoa. In this study, we evaluated the feasibility using rural domestic wastewater for agricultural irrigation, and analyzed dynamic changes of the microbial community structure and FLA populations in raw and treated wastewater, as well as the phyllosphere and rhizosphere of lettuce production sites that were irrigated with different water sources. The bacterial community dynamics were analyzed by terminal restriction fragment length polymorphism (T-RFLP). The bacterial community structures in the influent were similar to that in the effluent, while in some cases relative abundances varied significantly. The populations of Acanthamoeba spp. and Hartmannella vermiformis in the anaerobically treated wastewater were significantly higher than in the raw wastewater. The vegetables could harbor diverse amoebae, and the abundances of Acanthamoeba spp. and H. vermiformis in the rhizosphere were significantly higher than in the phyllosphere. Accordingly, our studies show insight into the distribution and dissemination of amoebae in wastewater treatment and irrigation practices.

Effects of nitrogen fertilization on the fate of high-risk antibiotic resistance genes in reclaimed water-irrigated soil and plants.

High-risk antibiotic resistance genes (ARGs) in reclaimed water-irrigated soil pose a potential threat to ecosystem and human health. Inorganic fertilization - including with nitrogen, a key ingredient in agricultural production - may affect the ARG profile in soil. However, little is known about nitrogen fertilization's influence on ARGs profiles in the soil-plant system. This study investigated the effects of different nitrogen fertilizer types (CO(NH2)2, NO3--N (NaNO3) and NH4+-N (NH4HCO3)) and different nitrogen fertilizer application rates (low, medium, high) on the distribution of high-risk ARGs in reclaimed water-irrigated soil and plants using quantitative PCR, high-throughput sequencing and metagenomic sequencing. Soil microcosms results revealed that nitrogen fertilization significantly affected the pattern of high-risk ARGs in soil, and also affected high-risk ARGs abundance and transfer capacity in plants. Compared with nitrogen fertilizer application rate, nitrogen fertilizer types significantly contributed to enhancing the soil resistome, with the order of CO(NH2)2 > NO3--N ≈ NH4+-N. The medium application of NO3--N and NH4+-N significantly reduced high-risk ARGs abundance in the leaf endophyte. Bacterial community mainly drove the variation of ARGs in nitrogen-fertilized soil-plant system, and class I integron and metal resistance genes (MRGs) also had direct effects on these high-risk ARGs. A similar high-risk ARGs pattern was also found in field plot experiments, and several dangerous pathogens were observed as the main high-risk ARGs potential hosts in nitrogen-fertilized soil. Based on an economic assessment, application of NH4+-N (NH4HCO3) could reduce costs by $1,312.83 ha-1 compared with NO3--N (NaNO3). These results showed that the more important role of nitrogen type might be an effective and economical way to control high-risk ARGs spread in soil-plant system under reclaimed water irrigation.

The introduction of influent sulfamethoxazole loads induces changes in the removal pathways of sulfamethoxazole in vertical flow constructed wetlands featuring hematite substrate.

High frequent detection of sulfamethoxazole (SMX) in wastewater cannot be effectively removed by constructed wetlands (CWs) with a traditional river sand substrate. The role of emerging substrate of hematite in promoting SMX removal and the effect of influent SMX loads remain unclear. The removal efficiency of SMX in hematite CWs was significantly higher than that in river sand CWs by 12.7-13.8% by improving substrate adsorption capacity, plant uptake and microbial degradation. With increasing influent SMX load, the removal efficiency of SMX in hematite CWs slightly increased, and the removal pathways varied significantly. The contribution of plant uptake was relatively small (< 0.1%) under different influent SMX loads. Substrate adsorption (37.8%) primarily contributed to SMX removal in hematite CWs treated with low-influent SMX. Higher influent SMX loads decreased the contribution of substrate adsorption, and microbial degradation (67.0%) became the main removal pathway. Metagenomic analyses revealed that the rising influent load increased the abundance of SMX-degrading relative bacteria and the activity of key enzymes. Moreover, the abundance of high-risk ARGs and sulfonamide resistance genes in hematite CWs did not increase with the increasing influent load. This study elucidates the potential improvements in CWs with hematite introduction under different influent SMX loads.

[Effects of Foliar Application of Silicon Fertilizers on Phyllosphere Bacterial Community and Functional Genes of Paddy Irrigated with Reclaimed Water].

Agricultural utilization of reclaimed water is considered to be an effective way to solve water shortage and reduce water environmental pollution. Silicon fertilizer can improve crop yield and quality and enhance crop resistance. The effect of foliar spray with silicon fertilizer on phyllosphere microbial communities remains lacking. In this study, a pot experiment was conducted to explore the effects of different types of silicon fertilizer on the composition and diversity of a phyllosphere bacterial community and the abundances of related functional genes in rice irrigated with reclaimed water. The results showed that Firmicutes, Proteobacteria, Actinobacteriota, Bacteroidota, and Verrucomicrobiota dominated the phyllosphere bacteria of rice. The relative abundance of Bacillus was higher than that of other treatments in RIS3. Reclaimed water irrigation significantly increased the relative abundances of the potential pathogens Pantoea and Enterobacter. The unclassified bacteria were also an important part of the bacterial community in the rice phyllosphere. Bacillus, Exiguobacterium, Aeromonas, and Citrobacter were significantly enriched by silicon fertilizer treatments. Functional prediction analysis showed that indicator species were mainly involved in metabolism and degradation functions, and the predicted functional groups of phyllosphere bacteria were attributed to chemoheterotrophy, aerobic chemoheterotrophy, nitrate reduction, and fermentation. Quantitative PCR results showed that AOA, AOB, and nifH genes were at low abundance levels in all treatments, and nirK genes was not significantly different among treatments. These results contribute to the in-depth understanding of the effects of foliar spray silicon fertilizer on the bacterial community structure and diversity of rice phyllosphere and provide a theoretical basis for the application of silicon fertilizer in reclaimed water irrigation agriculture.

Biodegradation of di-n-butyl phthalate by a newly isolated halotolerant Sphingobium sp.

A Gram-negative strain (TJ) capable of growing aerobically on mixed phthalate esters (PAEs) as the sole carbon and energy source was isolated from the Haihe estuary, Tianjin, China. It was identified as belonging to the Sphingobium genus on the basis of morphological and physiological characteristics and 16S rRNA and gyrb gene sequencing. The batch tests for biodegradation of di-n-butyl phthalate (DBP) by the Sphingobium sp. TJ showed that the optimum conditions were 30 °C, pH 7.0, and the absence of NaCl. Stain TJ could tolerate up to 4% NaCl in minimal salt medium supplemented with DBP, although the DBP degradation rates slowed as NaCl concentration increased. In addition, substrate tests showed that strain TJ could utilize shorter side-chained PAEs, such as dimethyl phthalate and diethyl phthalate, but could not metabolize long-chained PAEs, such as di-n-octyl phthalate, diisooctyl phthalate, and di-(2-ethyl-hexyl) phthalate. To our knowledge, this is the first report on the biodegradation characteristics of DBP by a member of the Sphingobium genus.

Response of Soil Microenvironment and Crop Growth to Cyclic Irrigation Using Reclaimed Water and Brackish Water.

The scarcity of freshwater resources has increased the use of nonconventional water resources such as brackish water, reclaimed water, etc., especially in water-scarce areas. Whether an irrigation cycle using reclaimed water and brackish water (RBCI) poses a risk of secondary soil salinization to crop yields needs to be studied. Aiming to find an appropriate use for different nonconventional water resources, pot experiments were conducted to study the effects of RBCI on soil microenvironments, growth, physiological characteristics and antioxidation properties of crops. The results showed the following: (1) compared to FBCI, the soil moisture content was slightly higher, without a significant difference, while the soil EC, sodium and chloride ions contents increased significantly under the RBCI treatment. With an increase in the reclaimed water irrigation frequency (Tri), the contents of EC, Na+ and Cl- in the soil decreased gradually, and the difference was significant; the soil moisture content also decreased gradually. (2) There were different effects of the RBCI regime on the soil's enzyme activities. With an increase in the Tri, the soil urease activity indicated a significant upward trend as a whole. (3) RBCI can alleviate the risk of soil salinization to some extent. The soil pH values were all below 8.5, and were without a risk of secondary soil alkalization. The ESP did not exceed 15 percent, and there was no possible risk of soil alkalization except that the ESP in soil irrigated by brackish water irrigation went beyond the limit of 15 percent. (4) Compared with FBCI, no obvious changes appeared to the aboveground and underground biomasses under the RBCI treatment. The RBCI treatment was conducive to increasing the aboveground biomass compared with pure brackish water irrigation. Therefore, short-term RBCI helps to reduce the risk of soil salinization without significantly affecting crop yield, and the irrigation cycle using reclaimed-reclaimed-brackish water at 3 g·L-1 was recommended, according to the experimental results.

Transcriptomic Sequencing Analysis on Key Genes and Pathways Regulating Cadmium (Cd) in Ryegrass (Lolium perenne L.) under Different Cadmium Concentrations

Perennial ryegrass (Lolium perenne L.) is an important forage grass and has the potential to be used in phytoremediation, while little information is available regarding the transcriptome profiling of ryegrass leaves in response to high levels of Cd. To investigate and uncover the physiological responses and gene expression characteristics of perennial ryegrass under Cd stress, a pot experiment was performed to study the transcriptomic profiles of ryegrass with Cd-spiked soils. Transcriptome sequencing and comparative analysis were performed on the Illumina RNA-Seq platform at different concentrations of Cd-treated (0, 50 and 500 mg·kg−1 soil) ryegrass leaves and differentially expressed genes (DEGs) were verified by RT-qPCR. The results show that high concentrations of Cd significantly inhibited the growth of ryegrass, while the lower concentrations (5 and 25 mg·kg−1) showed minor effects. The activity levels of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and malondialdehyde (MDA) increased in Cd-treated ryegrass leaves. We identified 1103 differentially expressed genes (DEGs) and profiled the molecular regulatory pathways of ryegrass leaves with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis in response to Cd stress. Cd stress significantly increased the membrane part, the metabolic process, the cellular process and catalytic activity. The numbers of unigenes related to signal transduction mechanisms, post-translational modification, replication, recombination and repair significantly increased. KEGG function annotation and enrichment analysis were performed based on DEGs with different treatments, indicating that the MAPK signaling pathway, the mRNA surveillance pathway and RNA transport were regulated significantly. Taken together, this study explores the effect of Cd stress on the growth physiology and gene level of ryegrass, thus highlighting significance of preventing and controlling heavy metal pollution in the future.

Does Short-Term Combined Irrigation Using Brackish-Reclaimed Water Cause the Risk of Soil Secondary Salinization?

Brackish water has to be used to irrigate crops for harvest due to the scarcity of freshwater resources. However, brackish water irrigation may cause secondary soil salinization. Whether the combined utilization of different non-conventional water resources could relieve the risk of secondary soil salinization has not been reported. In order to explore the safe and rational utilization of brackish water in areas where freshwater resources are scarce, a pot experiment was conducted to study the risk of secondary soil mixed irrigation and rotational irrigation using brackish water and reclaimed water or freshwater. The results indicated that: (1) Short-term irrigation using reclaimed water did not cause secondary soil salinization, although increasing soil pH value, ESP, and SAR. The indices did not exceed the threshold of soil salinization. (2) Compared with mixed irrigation using brackish-freshwater, the contents of soil exchangeable Ca2+, K+, and Mg2+ increased, and the content of soil exchangeable Na+ decreased under rotational irrigation using brackish-reclaimed water. In addition, the contents of soil exchangeable Na+ and Mg2+ under mixed irrigation or rotational irrigation were significantly lower, and the exchangeable K+ content of the soil was higher compared with brackish water irrigation. The exchangeable Ca2+ content under rotational irrigation was higher than that of brackish water irrigation, while the reverse was seen under mixed irrigation. (3) For different combined utilization modes of brackish water and reclaimed water, the ESP and SAR were the lowest under rotational irrigation, followed by mixed irrigation and brackish water irrigation. The ESP under brackish water treatment exceeded 15%, indicating a certain risk of salinization, while ESPs under other treatments were below 15%. Under mixed irrigation or rational irrigation using reclaimed-brackish water, the higher the proportion or rotational times of reclaimed water, the lower the risk of secondary soil salinization. Therefore, short-term combined irrigation using brackish water and reclaimed water will not cause the risk of secondary soil salinization, but further experiments need to verify or cooperate with other agronomic measures in long-term utilization.

[Effects of Soil Amendments on the Bacterial Diversity and Abundances of Pathogens and Antibiotic Resistance Genes in Rhizosphere Soil Under Drip Irrigation with Reclaimed Water].

Due to reclaimed water, irrigation can cause human health and environmental risks. Soil amendments are applied to reveal the abundance of pathogens and antibiotic resistance genes in rhizosphere soil irrigated by reclaimed water and to better understand the effects of environmental factors on the rhizosphere soil bacterial composition, which has guiding significance for the reasonable use of soil amendments. In this study, the effects of biochar, bioorganic fertilizer, humic acid, loosening soil essence, and corn vinasse on bacterial community diversity and certain gene abundances in rhizosphere soil under drip irrigation with reclaimed water were studied using high-throughput assays and quantitative PCR. The results showed that biochar significantly increased pH, organic matter, and total nitrogen contents in the rhizosphere soil. The corn vinasse significantly decreased soil pH and increased the contents of total nitrogen and total phosphorus but significantly increased the soil EC value (P<0.05). The effects of the five soil amendments on the α-diversity of rhizosphere bacteria were not significantly different. The bacterial community structure and diversity of rhizosphere bacteria were similar at different taxonomic levels, but their relative abundance was different. α-Proteobacteria, γ-Proteobacteria, Bacteroidia, Actinobacteria, Acidimicrobiia, and Anaerolineae were the dominant bacteria in all treatments. The dominant genera consisted of Pseudomonas, Sphingobium, Sphingomonas, Cellvibrio, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Flavobacterium, and Algoriphagus (relative abundance>1%). Correlation analysis of environmental factors showed that the composition of the rhizosphere bacterial community was strongly correlated with pH, EC, total nitrogen, and total phosphorus content. The abundances of pathogenic bacteria and antibiotic resistance genes were 103-107 copies·g-1 and 104-108 copies·g-1, respectively. There were significant differences in the detection levels of pathogens and antibiotic resistance genes. Bioorganic fertilizer, loosening soil essence, and corn vinasse significantly increased the abundances of some antibiotic resistance genes, whereas humic acid and corn vinasse significantly decreased the abundances of Pseudomonas syringae, Ralstonia solanacearum, and total coliforms (P<0.05). A significant correlation was found between pathogens (Arcobacter, Bacillus cereus, Pantoea agglomerans, and Fecal bacteroidetes) and antibiotic resistance genes (tetA, tetB, tetO, tetQ, sul1, ermB, and ermC). In conclusion, while monitoring pathogens and antibiotic resistance genes in the agricultural environment under reclaimed water irrigation, attention should be paid to the rational application of soil amendments to avoid exacerbating the spread of biological contamination.

Ryegrass (Lolium multiflorum L.) and Indian mustard (Brassica juncea L.) intercropping can improve the phytoremediation of antibiotics and antibiotic resistance genes but not heavy metals.

Lolium multiflorum and Brassica juncea display phytoremediation potential for heavy metals and antibiotics pollution. However, there is limited understanding of their function in removing combined pollutants (heavy metals, antibiotics and antibiotic resistance genes (ARGs)) under different cropping patterns. Sole cropping had little effect on heavy metals, but reduced antibiotics by 2.46%-84.88% and increased ARGs by 15.96%-33.82%. Intercropping was more beneficial to soil remediation and plant accumulation of L. multiflorum, and further increased the remediation of antibiotics by 2.38%-54.40%. Members of phyla (Actinobacteria, Bacteroidetes, and Proteobacteria) were mainly responsible for most antibiotics removal. Compared with sole cropping, intercropping reduced more ARGs abundance in rhizosphere soil for L. multiflorum (20.43%) and in bulk soil for B. juncea (23.22%). Mobile genetic elements (MGEs) played a significant role in the variation of ARGs. Further, sample type showed a higher indirect negative impact on ARGs by mainly affecting soil properties and bacterial community, and the co-occurrence between the bacterial community and ARGs in bulk soil was more complex than that in rhizosphere soil. Together these results suggest that phytoremediation of combined soil pollution was positive but limited, and intercropping resulted in enhanced removal efficiency when compared with sole cropping.

[Effects of Selected Biochars Application on the Microbial Community Structures and Diversities in the Rhizosphere of Water Spinach (Ipomoea aquatica Forssk.) Irrigated with Reclaimed Water].

The utilization of reclaimed water is one of the most important ways of alleviating the shortage of water resources for agricultural irrigation. As an effective disposal method for biomass waste, biochar has been widely used in the improvement and remediation of agricultural environments. However, few studies have been performed on the effects of biochar application on microbial community structures and pathogen abundances in rhizosphere soils irrigated with reclaimed water. Based on a pot experiment, high throughput sequencing technology and quantitative polymerase chain reaction (PCR) methods were used to investigate the effects of different biochars on the microbial community structure and diversity and pathogen abundance of rhizosphere soils irrigated with reclaimed water. The results showed that four different types of biochars had different effects on the soil nutrient status. Rice hull-derived biochar and rice straw-derived biochar resulted in significantly increased soil pH with reclaimed water irrigation, while peanut shell-derived biochar, rice straw-derived biochar, and wheat straw-derived biochar significantly increased EC values (P<0.05). The Sobs index, Shannon index, and Chao1 index of bacterial community in the rhizosphere soil significantly increased with rice straw-derived biochar treatment, while the Simpson index significantly decreased by adding peanut shell-derived biochar, rice hull-derived biochar, and wheat straw-derived biochar (P<0.05). There were differences in the relative abundances of bacterial communities in rhizosphere soils under different treatments. The dominant taxonomic groups at the phylum level were Proteobacteria, Actinobacteria, Chloroflexi, Bacteroidetes, and Acidobacteria. The dominant genera included Pseudomonas, Rheinheimera, Arthrobacter, Sphingomonas, and Aeromonas (relative abundance>5%). Redundancy (RDA) and heatmap analyses showed that the diversities and compositions of bacterial communities in rhizosphere soils in different treatments were closely related to soil EC values, organic matter, total nitrogen, and cadmium contents. Biochar application had no significant effect on the abundances of Aeromonas hydrophila and Bacillus cereus. Rice straw-derived biochar and peanut shell-derived biochar could significantly reduce the γ-Proteobacteria, while rice hull-derived biochar and wheat straw-derived biochar could significantly reduce the relative abundance of AOA (P<0.05). In conclusion, there were no obvious negative effects observed from reclaimed water irrigation on soil quality. Biochar application not only significantly improved the physicochemical properties of the soil, but also impacted the bacterial community structure and the abundance of the functional bacteria in the rhizosphere soil, which was closely related to the soil properties.

Investigation of bacterial diversity and pathogen abundances in gibel carp (Carassius auratus gibelio) ponds during a cyprinid herpesvirus 2 outbreak.

Cyprinid herpesvirus 2 (CyHV-2) infection is detrimental to gibel carp health and may result in severe economic loss in freshwater aquaculture. However, information regarding the interaction of this pathogen with the aquatic environment is scarce. In this study, quantitative polymerase chain reaction (qPCR) and high-throughput sequencing were used to determine the abundances of pathogens and bacterial community compositions in two aquaculture ponds in Jiangsu Province, China. The results indicate that the concentrations of six selected pathogens were higher in the water than in the sediment and that these concentrations peaked during disease outbreak. In total, 8,326 and 18,244 operational taxonomic units were identified from water and sediment samples, respectively. The dominant phyla were Proteobacteria, Actinobacteria, Cyanobacteria, Bacteroidetes, and Chlorobi in water samples and Proteobacteria, Firmicutes, Actinobacteria, Chloroflexi, and Bacteroidetes in sediment samples. Bacterial communities were similar at the phylum level in different ponds, although significant differences were observed at the genus level. In addition, bacterial diversity was associated with environmental factors (temperature, chemical oxygen demand, NO2- -N, NO3- -N, and NH4+ -N) in the pond where the outbreak occurred. Additionally, CyHV-2 abundance was positively correlated with dissolved oxygen levels and Aeromonas spp. abundance in pond water (p < .01). This study provides comprehensive insight into the mechanisms of interaction between potential pathogens and the freshwater environment of aquaculture ponds during CyHV-2 disease outbreaks. Furthermore, the results from this study can contribute to improvement of the aquatic environment and establishment of disease prevention and control measures.

[Effect of Different Reclaimed Water Irrigation Methods on Bacterial Community Diversity and Pathogen Abundance in the Soil-Pepper Ecosystem].

Reclaimed water is considered to be a reasonable and sustainable alternative water resource to improve water resource layout and mitigate the shortage of traditional water resources. Its use in irrigation will cause changes in the microbial community structure and opportunistic pathogen abundance in soils and crops, but few studies have been conducted on this subject. Peppers were used as the research subjects, and the treatments were direct irrigation of reclaimed water, mixed irrigation with freshwater and reclaimed water, rotated irrigation with freshwater and reclaimed water, with potable water irrigation as the control. The effects of different irrigation methods of reclaimed water on the soil physicochemical properties were analyzed through a pot experiment. Furthermore, changes in bacterial community and opportunistic pathogen abundance in pepper fruit and the rhizosphere under reclaimed water irrigation conditions were investigated based on high-throughput sequencing technology and quantitative PCR methods. The results showed that direct irrigation with reclaimed water increased soil EC and decreased soil pH. 16S rDNA high-throughput sequencing showed that Proteobacteria, Bacteroides, Actinobacteria, and Firmicutes were present in both pepper fruit and the rhizosphere at phylum level, and the most dominant genera (Pantoea, Pseudomonas, Sphingomonas, Sphingopyxis, Luteimonas, and Mariniflexile) were greatly affected by reclaimed water irrigation methods. Quantitative PCR results indicated that the influence of reclaimed water irrigation on the distribution and abundance of pathogenic bacteria in the soil-pepper system was different, and the abundance of Legionella spp. in pepper fruit and Pseudomonas syringae in the rhizosphere increased with reclaimed water irrigation. Our results indicated that the reclaimed water was suitable for agricultural irrigation, but different reclaimed water irrigation methods may introduce different degrees of microbial contamination. In addition, attention must be given to some opportunistic pathogens and phytopathogens.

[Effects of Different Levels of Irrigation with Reclaimed Water on Soil Enzyme Activity and Distribution of Thermotolerant Coliforms].

This study aimed to discern the effect of different levels of irrigation with reclaimed water on soil enzyme activities and on the distribution of thermotolerant coliforms. The effects of two irrigation water qualities (reclaimed water and tap water) and two irrigation levels (full irrigation and insufficient irrigation) on soil enzyme activity and distribution of thermotolerant coliforms were studied through indoor soil column irrigation. Results indicated that:① Reclaimed water irrigation increased soil urease and invertase activity compared with tap water under the same irrigation conditions. Compared with insufficient irrigation with reclaimed water, full irrigation with reclaimed water significantly increased soil invertase activity and deep soil catalase activity. ② At the same irrigation level, the number of thermotolerant coliforms in soil significantly increased with reclaimed water irrigation. The number of thermotolerant coliforms in soil under full irrigation with reclaimed water was significantly higher than with insufficient irrigation using reclaimed water. The number of thermotolerant coliforms in surface soil of the four irrigated treatments was significantly higher than in the deep soil layer, and with progression of irrigation, the number of thermotolerant coliforms in the 0-60 cm soil layer decreased. ③ There was a significant positive correlation between the quantity of thermotolerant coliforms and the activity of soil urease and invertase, and between the quantity of thermotolerant coliforms and the activity of soil catalase. Reclaimed water therefore promoted enzyme activity that can promote transformation of C and N in soil. Reasonable irrigation with reclaimed water can therefore effectively control the number of thermotolerant coliforms in soil.

Bacterial communities and potential waterborne pathogens within the typical urban surface waters.

Waterborne pathogens have attracted a great deal of attention in the public health sector over the last several decades. However, little is known about the pathogenic microorganisms in urban water systems. In this study, the bacterial community structure of 16 typical surface waters in the city of Beijing were analyzed using Illumina MiSeq high-throughput sequencing based on 16S rRNA gene. The results showed that Bacteroidetes, Proteobacteria and Actinobacteria were the dominant groups in 16 surface water samples, and Betaproteobacteria, Alphaproteobacteria, Flavobacteriia, Sphingobacteriia and Actinobacteria were the most dominant classes. The dominant genus across all samples was Flavobacterium. In addition, fifteen genus level groups of potentialy pathogenic bacteria were detected within the 16 water samples, with Pseudomonas and Aeromonas the most frequently identified. Spearman correlation analysis demonstrated that richness estimators (OTUs and Chao1) were correlated with water temperature, nitrate and total nitrogen (p < 0.05), while ammonia-nitrogen and total nitrogen were significantly correlated with the percent of total potential pathogens (p ≤ 0.05). These results could provide insight into the ecological function and health risks of surface water bacterial communities during the process of urbanization.

[Influence of Mirabilis jalapa Linn. Growth on the Microbial Community and Petroleum Hydrocarbon Degradation in Petroleum Contaminated Saline-alkali Soil].

In order to explore the effect of Mirabilis jalapa Linn. growth on the structure characteristics of the microbial community and the degradation of petroleum hydrocarbon (TPH) in the petroleum-contaminated saline-alkali soil, Microbial biomass and species in the rhizosphere soils of Mirabilis jalapa Linn. in the contaminated saline soil were studied with the technology of phospholipid fatty acids (PLFAs) analysis. The results showed that comparing to CK soils without Mirabilis jalapa Linn., the ratio of PLFAs species varied were 71. 4%, 69. 2% and 33. 3% in the spring, summer and autumn season, respectively. In addition, there was distinct difference of the biomasses of the microbial community between the CK and rhizosphere soils and among the difference seasons of growth of Mirabilis jalapa Linn.. Compare to CK soil, the degradation rates of total petroleum hydrocarbon (TPH) was increased by 47. 6%, 28. 3%, and 18. 9% in spring, summer, and autumn rhizosphere soils, respectively. Correlation analysis was used to determine the correlation between TPH degradation and the soil microbial community. 77. 8% of the total soil microbial PLFAs species showed positive correlation to the TPH degradation (the correlation coefficient r > 0), among which, 55. 6% of PLFAs species showed high positive correlation(the correlation coefficient was r≥0. 8). In addition, the relative content of SAT and MONO had high correlation with TPH degradation in the CK sample soils, the corelation coefficient were 0. 92 and 0. 60 respectively; However, the percent of positive correlation was 42. 1% in the rhizosphere soils with 21. 1% of them had high positive correlation. The relative content of TBSAT, MONO and CYCLO had moderate or low correlation in rhizosphere soils, and the correlation coefficient were 0. 56, 0. 50, and 0. 07 respectively. Our study showed that the growth of mirabilis Mirabilis jalapa Linn. had a higher influence on the species and biomass of microbial community in the rhizosphere soils, and the results will provide a basis theory for the research of phytoremediation petroleum contaminated saline soil.