Biolog Eco Research Articles

The response of soil microbial community to application of organic amendment to saline land.

The salinization of coastal soils is a primary cause of global land degradation. The aim of this study was to evaluate the effect of organic amendment on the soil microbial community within a saline gradient. The study was designed with five levels of electrical conductivity (EC): 0.33, 0.62, 1.13, 1.45 and 2.04 ds m-1. By conducting indoor potted plant experiments, determine the effects of applying microbial organic fertilizer on the physicochemical properties of soil and the structure of soil microbial communities under different salinity concentrations. Compared with the control, higher OM content, total N, and higher crop biomass were observed in samples with organic amendment at the same salinity level. At the same salinity levels, the mean bacterial activity (AUC) and the mean number of substrates were higher than in the soil without organic amendment according to analyses by means of Biolog ECO MicroPlate. The results of canonical correspondence analysis indicate that after the application of organic amendments, the composition of loam and clay replaces soil pH, and aboveground biomass replaces root biomass as key indicators closely monitoring Community level physiological profiling (CLPP). In soil with the same salinity level, the application of microbial organic fertilizer led to an increase in the proportion of Actinobacteriota and a decrease in the proportion of Chloroflexi. In 0.3dS m-1 soil, the abundance of actinomycetes increased from 23% to 27% after application of microbial organic fertilizer, while the abundance of basidiomycetes decreased from 20% to 6%. In addition, after the application of microbial organic fertilizer, RB41, blastococcus and solirubrobacter significantly increased, while Melothermus and Herpetosiphon significantly decreased. This study provides a strong theoretical basis for using microbial organic fertilizers to improve saline-alkali soil.

Microbial Community Structure and Metabolic Function in the Venom Glands of the Predatory Stink Bug, Picromerus lewisi (Hemiptera: Pentatomidae).

The predatory stink bug, Picromerus lewisi (Hemiptera: Pentatomidae), is an important and valuable natural enemy of insect pests in their ecosystems. While insects are known to harbor symbiotic microorganisms, and these microbial symbionts play a crucial role in various aspects of the host's biology, there is a paucity of knowledge regarding the microbiota present in the venom glands of P. lewisi. This study investigated the venom glands of adult bugs using both traditional in vitro isolation and cultural methods, as well as Illumina high-throughput sequencing technology. Additionally, the carbon metabolism of the venom gland's microorganisms was analyzed using Biolog ECO metabolic phenotyping technology. The results showed 10 different culturable bacteria where the dominant ones were Enterococcus spp. and Lactococcus lactis. With high-throughput sequencing, the main bacterial phyla in the microbial community of the venom glands of P. lewisi were Proteobacteria (78.1%) and Firmicutes (20.3%), with the dominant bacterial genera being Wolbachia, Enterococcus, Serratia, and Lactococcus. At the fungal community level, Ascomycota accounted for the largest proportion (64.1%), followed by Basidiomycota (27.6%), with Vishniacozyma, Cladosporium, Papiliotrema, Penicillium, Fusarium, and Aspergillus as the most highly represented fungal genera. The bacterial and fungal community structure of the venom glands of P. lewisi exhibited high species richness and diversity, along with a strong metabolism of 22 carbon sources. Functional prediction indicated that the primary dominant function of P. lewisi venom-gland bacteria was metabolism. The dominant eco-functional groups of the fungal community included undefined saprotroph, fungal parasite-undefined saprotroph, unassigned, endophyte-plant pathogen, plant pathogen-soil saprotroph-wood saprotroph, animal pathogen-endophyte-plant pathogen-wood saprotroph, plant pathogen, and animal pathogen-endophyte-epiphyte-plant pathogen-undefined saprotroph. These results provide a comprehensive characterization of the venom-gland microbiota of P. lewisi and demonstrate the stability (over one week) of the microbial community within the venom glands. This study represents the first report on the characterization of microbial composition from the venom glands of captive-reared P. lewisi individuals. The insights gained from this study are invaluable for future investigations into P. lewisi's development and the possible interactions between P. lewisi's microbiota and some Lepidopteran pests.

Mixed planting of subtropical Chinese fir in South China improves microbial carbon source metabolism and functional diversity through the accumulation of nutrients from soil aggregates.

Soil microbial functional traits are key indicators of soil microbial ecological traits; however, how mixing patterns of Chinese fir and broadleaved trees drive soil microbial functional trait variation at the aggregate scale and how soil microbial functional traits are linked to soil fertility factors have largely not been determined. In this study, soil from the 0-20 cm depths in three Chinese fir plantations was collected, and the soil samples were separated into >2 mm (large macro-aggregate), 0.25-2 mm (macro-aggregate) and <0.25 mm (micro-aggregate) by complying with an optimal moisture sieving procedure. The metabolic activities, functional diversity and different carbon sources utilization characteristics of the soil microorganisms were determined by the Biolog Eco microplate method. In all Chinese fir plantations, micro-aggregates (<0.25 mm) consistently exhibited the highest levels of microbial metabolic activity, a more uniform carbon source utilisation capacity, and the highest microbial diversity. Micro-aggregates also showed elevated levels of soil organic carbon (OC), total nitrogen (TN), total phosphorus (TP), and higher ratios of C/N and C/P compared to large macro-aggregates and macro-aggregates, indicating that micro-aggregates contain more resources available to soil microorganisms. Soil OC, TN, and TP content were enhanced by integration with Michelia macclurei, suggesting that this combination promotes relatively favourable soil conditions for microbial growth and multiplication. This, in turn, promotes microbial metabolic activity. Furthermore, redundancy and correlation analyses showed that soil OC, TN, and TP were identified as principal determinants of soil microbial functional properties in Chinese fir plantations. In summary, mixed cultivation and aggregate size influenced microbial functional properties via soil nutrient alterations. Consequently, adopting a mixed cultivation approach of Chinese fir and broadleaved trees is advocated in the subtropical regions of Guangxi. Employing a diversity of tree species, including M. macclurei, is recommended for optimal soil quality preservation.

Response of petroleum-contaminated soil to chemical oxidation combined with biostimulation

In this study, a microcosm experiment was conducted to investigate the effects of Na2S2O8 preoxidation combined with biostimulation on petroleum-contaminated soil remediation. The response of microbial community during this process was explored using BIOLOG ECO microplate carbon utilization method and 16 s rDNA high-throughput sequencing. The results showed that use of 10 mg/g Na2S2O8 removed 19.8 % of the petroleum hydrocarbons, reduced soil biotoxicity and did not affect soil microbial activity compared to other concentrations. Therefore, sodium persulfate of ca. 10 mg/g was used to oxidize petroleum in soil before the biostimulation experiment with organic and inorganic fertilizers. Our finding showed that the content of total petroleum hydrocarbons (TPHs) in soil was reduced by 43.3 % in inorganic fertilizer treatment after 60 days. The results of BIOLOG ECO microplate carbon utilization analysis and 16 S rDNA high-throughput sequencing further confirmed that biostimulation quickly restored the microbial activities in oxidant treated soil. The main marker bacteria in chemical oxidation combined with biostimulation remediation were Arthrobacter and Paenarthrobacter, and their relative abundances were both significantly negatively correlated with the content of petroleum hydrocarbons in soil.

Tea and Pleurotus ostreatus intercropping modulates structure of soil and root microbial communities.

Intercropping with Pleurotus ostreatus has been demonstrated to increase the tea yield and alleviate soil acidification in tea gardens. However, the underlying mechanisms remain elusive. Here, high-throughput sequencing and Biolog Eco analysis were performed to identify changes in the community structure and abundance of soil microorganisms in the P. ostreatus intercropped tea garden at different seasons (April and September). The results showed that the soil microbial diversity of rhizosphere decreased in April, while rhizosphere and non-rhizosphere soil microbial diversity increased in September in the P. ostreatus intercropped tea garden. The diversity of tea tree root microorganisms increased in both periods. In addition, the number of fungi associated with organic matter decomposition and nutrient cycling, such as Penicillium, Trichoderma, and Trechispora, was significantly higher in the intercropped group than in the control group. Intercropping with P. ostreatus increased the levels of total nitrogen (TN), total phosphorus (TP), and available phosphorus (AP) in the soil. It also improved the content of secondary metabolites, such as tea catechins, and polysaccharides in tea buds. Microbial network analysis showed that Unclassified_o__Helotiales, and Devosia were positively correlated with soil TN and pH, while Lactobacillus, Acidothermus, and Monascus were positively correlated with flavone, AE, and catechins in tea trees. In conclusion, intercropping with P. ostreatus can improve the physical and chemical properties of soil and the composition and structure of microbial communities in tea gardens, which has significant potential for application in monoculture tea gardens with acidic soils.

Acidification induce chemical and microbial variation in tea plantation soils and bacterial degradation of the key acidifying phenolic acids.

Camellia sinensis is an important economic plant grown in southern subtropical hilly areas, especially in China, mainly for the production of tea. Soil acidification is a significant cause of the reduction of yield and quality and continuous cropping obstacles in tea plants. Therefore, chemical and microbial properties of tea growing soils were investigated and phenolic acid-degrading bacteria were isolated from a tea plantation. Chemical and ICP-AES investigations showed that the soils tested were acidic, with pH values of 4.05-5.08, and the pH negatively correlated with K (p < 0.01), Al (p < 0.05), Fe and P. Aluminum was the highest (47-584mg/kg) nonessential element. Based on high-throughput sequencing, a total of 34 phyla and 583 genera were identified in tea plantation soils. Proteobacteria and Acidobacteria were the main dominant phyla and the highest abundance of Acidobacteria was found in three soils, with nearly 22% for the genus Gp2. Based on the functional abundance values, general function predicts the highest abundance, while the abundance of amino acids and carbon transport and metabolism were higher in soils with pH less than 5. According to Biolog Eco Plate™ assay, the soil microorganisms utilized amino acids well, followed by polymers and phenolic acids. Three strains with good phenolic acid degradation rates were obtained, and they were identified as Bacillus thuringiensis B1, Bacillus amyloliquefaciens B2 and Bacillus subtilis B3, respectively. The three strains significantly relieved the inhibition of peanut germination and growth by ferulic acid, p-coumaric acid, p-hydroxybenzoic acid, cinnamic acid, and mixed acids. Combination of the three isolates showed reduced relief of the four phenolic acids due to the antagonist of B2 against B1 and B3. The three phenolic acid degradation strains isolated from acidic soils display potential in improving the acidification and imbalance in soils of C. sinensis.

Analysis on metabolic functions of rhizosphere microbial communities of Pinus massoniana provenances with different carbon storage by Biolog Eco microplates.

Rhizosphere microorganisms are influenced by vegetation. Meanwhile, they respond to vegetation through their own changes, developing an interactive feedback system between microorganisms and vegetation. However, it is still unclear whether the functional diversity of rhizosphere soil microorganisms varies with different carbon storage levels and what factors affect the functional diversity of rhizosphere soil microorganisms. In this study, the Biolog-Eco microplate technique was used to analyze the metabolic diversity of carbon source of rhizosphere soil microorganisms from 6 Pinus massoniana provenances with three levels of high, medium and low carbon storage. The results showed that the average well color development(AWCD) value of rhizosphere microorganisms was significantly positive correlated with carbon storage level of Pinus massoniana (p < 0.05). The AWCD value, Simpson and Shannon diversity of high carbon sequestrance provenances were 1.40 (144h incubation) 0.96 and 3.24, respectively, which were significantly higher (p < 0.05) than those of other P. massoniana provenances. The rhizosphere microbial AWCD, Shannon and Simpson diversity of the 6 provenances showed the same variation trend (SM>AY>QJ>SX>HF>SW). Similarly, microbial biomass carbon (MBC) content was positively correlated with carbon storage level, and there were significant differences among high, medium and low carbon storage provenances. The PCA results showed that the differences in the carbon source metabolism of rhizosphere microorganisms were mainly reflected in the utilization of amino acids, carboxylic acids and carbohydrates. Pearson correlation analysis showed that soil organic carbon (SOC), total nitrogen (TN) and pH were significantly correlated with rhizosphere AWCD (p < 0.05). Soil properties are important factors affecting rhizosphere microbial carbon source metabolism. The study confirmed that the microorganisms of high carbon storage provenances had relatively high carbon metabolic activity. Among them, the carbon metabolic activity of rhizosphere microorganisms of SM provenance was the highest, which was the preferred provenances in effective ecological service function.

Phyllospheric microbial community structure and carbon source metabolism function in tobacco wildfire disease.

The phyllospheric microbial composition of tobacco plants is influenced by multiple factors. Disease severity level is one of the main influencing factors. This study was designed to understand the microbial community in tobacco wildfire disease with different disease severity levels. Tobacco leaves at disease severity level of 1, 5, 7, and 9 (L1, L5, L7, and L9) were collected; both healthy and diseased leaf tissues for each level were collected. The community structure and diversity in tobacco leaves with different disease severity levels were compared using high-throughput technique and Biolog Eco. The results showed that in all healthy and diseased tobacco leaves, the most dominant bacterial phylum was Proteobacteria with a high prevalence of genus Pseudomonas; the relative abundance of Pseudomonas was most found at B9 diseased samples. Ascomycota represents the most prominent fungal phylum, with Blastobotrys as the predominant genus. In bacterial communities, the Alpha diversity of healthy samples was higher than that of diseased samples. In fungal community, the difference in Alpha diversity between healthy and diseased was not significant. LEfSe analysis showed that the most enriched bacterial biomarker was unclassified_Gammaproteobacteria in diseased samples; unclassified_Alcaligenaceae were the most enrich bacterial biomarker in healthy samples. FUNGuild analysis showed that saprotroph was the dominated mode in health and lower diseased samples, The abundance of pathotroph-saprotroph and pathotroph-saprotroph-symbiotroph increases at high disease levels. PICRUSt analysis showed that the predominant pathway was metabolism function, and most bacterial gene sequences seem to be independent of the disease severity level. The Biolog Eco results showed that the utilization rates of carbon sources decrease with increasing disease severity level. The current study revealed the microbial community's characteristic of tobacco wildfire disease with different disease severity levels, providing scientific references for the control of tobacco wildfire disease.

Characteristics of Epicoccum latusicollum as revealed by genomic and metabolic phenomic analysis, the causal agent of tobacco Epicoccus leaf spot.

Epicoccum latusicollum is a fungus that causes a severe foliar disease on flue-cured tobacco in southwest China, resulting in significant losses in tobacco yield and quality. To better understand the organism, researchers investigated its optimal growth conditions and metabolic versatility using a combination of traditional methods and the Biolog Phenotype MicroArray technique. The study found that E. latusicollum exhibited impressive metabolic versatility, being able to metabolize a majority of carbon, nitrogen, sulfur, and phosphorus sources tested, as well as adapt to different environmental conditions, including broad pH ranges and various osmolytes. The optimal medium for mycelial growth was alkyl ester agar medium, while oatmeal agar medium was optimal for sporulation, and the optimum temperature for mycelial growth was 25°C. The lethal temperature was 40°C. The study also identified arbutin and amygdalin as optimal carbon sources and Ala-Asp and Ala-Glu as optimal nitrogen sources for E. latusicollum. Furthermore, the genome of E. latusicollum strain T41 was sequenced using Illumina HiSeq and Pacific Biosciences technologies, with 10,821 genes predicted using Nonredundant, Gene Ontology, Clusters of Orthologous Groups, Kyoto Encyclopedia of Genes and Genomes, and SWISS-PROT databases. Analysis of the metabolic functions of phyllosphere microorganisms on diseased tobacco leaves affected by E. latusicollum using the Biolog Eco microplate revealed an inability to efficiently metabolize a total of 29 carbon sources, with only tween 40 showing some metabolizing ability. The study provides new insights into the structure and function of phyllosphere microbiota and highlights important challenges for future research, as well as a theoretical basis for the integrated control and breeding for disease resistance of tobacco Epicoccus leaf spot. This information can be useful in developing new strategies for disease control and management, as well as enhancing crop productivity and quality.

Copper-oxide nanoparticles exert persistent changes in the structural and functional microbial diversity: A 60-day mesocosm study of zinc-oxide and copper-oxide nanoparticles in the soil-microorganism-nanoparticle system

Recent advances in nanotechnology and development of nanoformulation methods, has enabled the emergence of precision farming – a novel farming method that involves nanopesticides and nanoferilizers. Zinc-oxide nanoparticles serve as a Zn source for plants, but they are also used as nanocarriers for other agents, whereas copper-oxide nanoparticles possess antifungal activity, but in some cases may also serve as a micronutrient providing Cu ions. Excessive application of metal-containing agents leads to their accumulation in soil, where they pose a threat to non-target soil organisms. In this study, soils obtained from the environment were amended with commercial zinc-oxide nanoparticles: Zn-OxNPs(10−30), and newly-synthesized copper-oxide nanoparticles: Cu-OxNPs(1−10). Nanoparticles (NPs) in 100 and 1000 mg kg-1 concentrations were added in separate set-ups, representing a soil-microorganism-nanoparticle system in a 60-day laboratory mesocosm experiment. To track environmental footprint of NPs on soil microorganisms, a Phospholipd Fatty Acid biomarker analysis was employed to study microbial community structure, whereas Community-Level Physiological Profiles of bacterial and fungal fractions were measured with Biolog Eco and FF microplates, respectively. The results revealed a prominent and persistent effects exerted by copper-containing nanoparticles on non-target microbial communities. A severe loss of Gram-positive bacteria was observed in conjunction with disturbances in bacterial and fungal CLPPs. These effects persisted till the end of a 60-day experiment, demonstrating detrimental rearrangements in microbial community structure and functions. The effects imposed by zinc-oxide NPs were less pronounced. As persistent changes were observed for newly synthesized Cu-containing NPs, this work stresses the need for obligatory testing of nanoparticle interactions with non-target microbial communities in long-term experiments, especially during the approval procedures of novel nano-substances. It also underlines the role of in-depth physical and chemical studies of NP-containing agents, which may be tweaked to mitigate the unwanted behavior of such substances in the environment and preselect their beneficial characteristics.

New insights into the effects of antibiotics and copper on microbial community diversity and carbon source utilization.

Residual antibiotics (ABs) and heavy metals (HMs) are continuously released from soil, reflecting their intensive use and contamination of water and soil, posing an environmental problem of great concern. Relatively few studies exist of the functional diversity of soil microorganisms under the combined action of ABs and HMs. To address this deficiency, BIOLOG ECO microplates and the Integrated Biological Responses version 2 (IBRv2) method were used to comprehensively explore the effects of single and combined actions of copper (Cu) and enrofloxacin (ENR), oxytetracycline (OTC), and sulfadimidine (SM2) on the soil microbial community. The results showed that the high concentration (0.80mmol/kg) compound group had a significant effect on average well color development (AWCD) and OTC showed a dose-response relationship. The results of IBRv2 analysis showed that the single treatment group of ENR or SM2 had a significant effect on soil microbial communities, and the IBRv2 of E1 was 5.432. Microbes under ENR, SM2, and Cu stress had more types of available carbon sources, and all treatment groups were significantly more enriched with microorganisms having D-mannitol and L-asparagine as carbon sources. This study confirms that the combined effects of ABs and HMs can inhibit or promote the function of soil microbial communities. In addition, this paper will provide new insights into IBRv2 as an effective method to evaluate the impacts of contaminants on soil health.

Dynamic responses of carbon metabolism of sediment microbial communities to Ag nanoparticles: Effects of the single and repeated exposure scenarios

The widespread use of nanosilver will inevitably lead to their release into aquatic environment, threating the health of freshwater ecosystem. The toxic effects of silver nanoparticles (AgNPs) on sediment microbial diversity, community composition, and functional enzyme activity are well established, while little is known about how sediment microbes dynamically respond to the stress of different AgNPs exposure scenarios. Herein, microcosm experiments were performed to investigate the impacts of repeated (1 mg/L, applied every 6 days for 10 times) and single (10 mg/L) exposure scenarios of AgNPs on the specific functions of sediment microbes (5–60 days). The carbon metabolism of sediment microbial communities was measured using BIOLOG ECO microplates, and carbon metabolic rate and functional diversity indices were calculated. Compared to control group, the maximum carbon source utilization capacity of the microbial community increased by 6.6 and 15.4 % in the single and repeated exposure group, respectively. And the metabolic rates of sediment microorganisms were significant increased by 6.1 % in the repeated exposure group, which suggested that repetitive low-dosing of AgNPs induce a larger alteration of both capacity and rate of microbial carbon metabolism. Notably, different AgNPs exposure scenarios resulted in a shift in the carbon source preference of the microorganisms. After exposure for 60 days, compared with the controls, the ability to utilize polymers was significantly increased by 51.5 and 21.7 % in the single and repeated exposure groups, respectively, and decreased by 33.7 and 10.5 % in the utilization of miscellaneous, both exhibiting significant differences (P < 0.05), implying that AgNPs exposure scenarios affected the microbial-mediated carbon cycling processes in sediment. These results highlight that different exposure scenarios of AgNPs have different effects on the carbon metabolism capacity of microbial communities, thus affecting the carbon cycling processes in which microorganisms are involved.

Effects of the Desiccation Duration on the Dynamic Responses of Biofilm Metabolic Activities to Rewetting

Global climate changes have increased the duration and frequency of river flow interruption, affecting the physical and community structure of benthic biofilms. However, the dynamic responses of biofilm metabolism during the dry-wet transition remain poorly understood. Herein, the dynamic changes in biofilm metabolic activities were investigated through mesocosm experiments under short-term (25 day) and long-term drought (90 day), followed by a 20 day rewetting. The biofilm ecosystem metabolism, as measured by gross primary production and community respiration, was significantly inhibited and turned heterotrophic during the desiccation phase and then recovered, becoming autotrophic during the rewetting period regardless of the desiccation periods due to the high resilience of the autotrophic community. However, long-term drought decreased the recovery rate of the ecosystem metabolism and also caused irreparable damage to the biofilm carbon metabolism, measured using Biolog Eco Plates. Specifically, the recovery of the total carbon metabolic activity is related to the specific carbon source utilized by biofilm microorganisms, such as polymers, carbohydrates, and carboxylic acids. However, the divergent changes of amino acids caused the failure of the total carbon metabolism in long-term drought treatments to recover to the control level even after 20 days of rewetting. This research provides direct evidence that the increased duration of non-flow periods affects biofilm-mediated carbon biogeochemical processes.

In-situ sludge reduction based on Mn2+-catalytic ozonation conditioning: Feasibility study and microbial mechanisms

To improve the sludge conditioning efficiency without increasing the ozone dose, an in-situ sludge reduction process based on Mn2+-catalytic ozonation conditioning was proposed. Using ozone conditioning alone as a control, a lab-scale sequencing batch reactor coupled with ozonated sludge recycle was evaluated for its operating performance at an ozone dose of 75 mg O3/g VSS and 1.5 mmol/L Mn2+ addition. The results showed a 39.4% reduction in MLSS and an observed sludge yield of 0.236 kg MLSS/kg COD for the O3+Mn2+ group compared to the O3 group (15.3% and 0.292 kg MLSS/kg COD), accompanied by better COD, NH4+-N, TN and TP removal, improved effluent SS and limited impact on excess sludge properties. Subsequently, activity tests, BIOLOG ECO microplates and 16S rRNA sequencing were applied to elucidate the changing mechanisms of Mn2+-catalytic ozonation related to microbial action: (1) Dehydrogenase activity reached a higher peak. (2) Microbial utilization of total carbon sources had an elevated effect, up to approximately 18%, and metabolic levels of six carbon sources were also increased, especially for sugars and amino acids most pronounced. (3) The abundance of Defluviicoccus under the phylum Proteobacteria was enhanced to 12.0% and dominated in the sludge, they had strong hydrolytic activity and metabolic capacity. Denitrifying bacteria of the genus Ferruginibacter also showed an abundance of 7.6%, they contributed to the solubilization and reduction of sludge biomass. These results could guide researchers to further reduce ozonation conditioning costs, improve sludge management and provide theoretical support.

Effects of Straw Returning on Soil Chemical Properties and Microbial Community Diversity under the Rice-Crayfish Integrated System

This study presents an investigation of soil chemical properties and microbial community diversity by Biolog ECO analysis in a seven-year field experiment using winter flooded fallow + no straw returning (W), winter flooded fallow + straw returning (WS), and winter flooded fallow + straw returning + crayfish farming (WSC) at soil depths of 0–10 cm and 10–20 cm. Compared with the WS treatment, the WSC treatment had significantly higher total organic carbon (TOC) content in the 0–10 cm layer as well as greater available nitrogen (AN) and total nitrogen (TN) contents and acid phosphatase and sucrase activities in the 10–20 cm layer, while the pH value, total reducing substances, and Fe2+ content in the 0–20 cm layer were considerably lower. The WSC treatment improved the microbial species abundance in the 10–20 cm layer and the utilization rate of carbon sources in the 0–20 cm layer compared with the WS treatment. The soil microbial species abundance, microbial community diversity, and utilization rate of carbon sources in all of the layers examined were significantly higher in the WSC treatment than in the W treatment. The results indicate that straw returning under the rice-crayfish integrated system improves the contents of TOC, TN, and AN, decreases reducing substances properties, increases acid phosphatase and sucrase activities, and improves microbial community functional diversity, thereby contributing to the improvement of soil quality and the long-term sustainable development of the rice-crayfish integrated system.

Effects of Specific Surface Area of Artificial Carriers on Carbon Metabolism Activity of Biofilm

Biofilm technology has been applied to polluted water treatment. The functional characteristics of biofilm play key roles in the decontamination efficiency and are affected by the physicochemical properties of the carrier surface. Previous studies reported that the larger the specific surface area of the artificial carrier, the more biofilms there are attached to its surface. However, the coupling relationship between the metabolic activity of biofilm and the specific surface area of carrier remained limitedly studied. Herein, five carriers with a gradient of specific surface areas were selected to incubate biofilms in two low-pollution rivers for 44 days, and a BIOLOG ECO microplate was used to analyze the metabolic activity of biofilm. Results demonstrated that the carbon metabolic capacity of biofilm fluctuates with the increase in the specific surface area. In Xuanwu Lake, the carbon metabolism rate of biofilm demonstrated a linear correlation with a specific surface area. In Donghu Lake, there was a trend of a single hump because of mass transfer limitation. In different water bodies, the carbon metabolic activity of biofilm demonstrated different trends with the change of specific surface area, and mass transfer limitation should be considered. For the classified carbon sources, there was a threshold (with Xuanwu Lack of 7500 m2/m3 and Donghu Lack of 5000 m2/m3) to make the biofilm achieve the best utilization ability of classified carbon sources. Nevertheless, the change of carriers’ specific surface area had little effect on the community structure of biofilm.

Effects of salinomycin and ethanamizuril on the three microbial communities in vivo and in vitro.

The fate of a drug is not only the process of drug metabolism in vivo and in vitro but also the homeostasis of drug-exposed microbial communities may be disturbed. Anticoccidial drugs are widely used to combat the detrimental effects of protozoan parasites in the poultry industry. Salinomycin and ethanamizuril belong to two different classes of anticoccidial drugs. The effect of salinomycin and ethanamizuril on the microbiota of cecal content, manure compost, and soil remains unknown. Our results showed that although both salinomycin and ethanamizuril treatments suppressed some opportunistic pathogens, they failed to repair the great changes in chicken cecal microbial compositions caused by coccidia infection. Subsequently, the metabolite5 profiling of cecal content by LC-MS/MS analyses confirmed the great impact of coccidia infection on chicken cecum and showed that histidine metabolism may be the main action pathway of salinomycin, and aminoacyl tRNA biosynthesis may be the major regulatory mechanism of ethanamizuril. The microbial community of manure compost showed a mild response to ethanamizuril treatment, but ethanamizuril in soil could promote Actinobacteria reproduction, which may inhibit other taxonomic bacteria. When the soil and manure were exposed to salinomycin, the Proteobacteria abundance of microbial communities showed a significant increase, which suggested that salinomycin may improve the ability of the microbiota to utilize carbon sources. This hypothesis was confirmed by a BIOLOG ECO microplate analysis. In the animal model of coccidia infection, the treatment of salinomycin and ethanamizuril may reconstruct a new equilibrium of the intestinal microbiota. In an in vitro environment, the effect of ethanamizuril on composting and soil microbiota seems to be slight. However, salinomycin has a great impact on the microbial communities of manure composting and soil. In particular, the promoting effect of salinomycin on Proteobacteria phylum should be further concerned. In general, salinomycin and ethanamizuril have diverse effects on various microbial communities.

Spatio-Temporal Variation in the Phyllospheric Microbial Biodiversity of Alternaria Alternata-Infected Tobacco Foliage.

Phyllospheric microbial composition of tobacco (Nicotiana tabacum L.) is contingent upon certain factors, such as the growth stage of the plant, leaf position, and cultivar and its geographical location, which influence, either directly or indirectly, the growth, overall health, and production of the tobacco plant. To better understand the spatiotemporal variation of the community and the divergence of phyllospheric microflora, procured from healthy and diseased tobacco leaves infected by Alternaria alternata, the current study employed microbe culturing, high-throughput technique, and BIOLOG ECO. Microbe culturing resulted in the isolation of 153 culturable fungal isolates belonging to 33 genera and 99 bacterial isolates belonging to 15 genera. High-throughput sequencing revealed that the phyllosphere of tobacco was dominantly colonized by Ascomycota and Proteobacteria, whereas, the most abundant fungal and bacterial genera were Alternaria and Pseudomonas. The relative abundance of Alternaria increased in the upper and middle healthy groups from the first collection time to the third, whereas, the relative abundance of Pseudomonas, Sphingomonas, and Methylobacterium from the same positions increased during gradual leaf aging. Non-metric multi-dimensional scaling (NMDs) showed clustering of fungal communities in healthy samples, while bacterial communities of all diseased and healthy groups were found scattered. FUNGuild analysis, from the first collection stage to the third one in both groups, indicated an increase in the relative abundance of Pathotroph-Saprotroph, Pathotroph-Saprotroph-Symbiotroph, and Pathotroph-Symbiotroph. Inclusive of all samples, as per the PICRUSt analysis, the predominant pathway was metabolism function accounting for 50.03%. The average values of omnilog units (OUs) showed relatively higher utilization rates of carbon sources by the microbial flora of healthy leaves. According to the analysis of genus abundances, leaf growth and leaf position were the important drivers of change in structuring the microbial communities. The current findings revealed the complex ecological dynamics that occur in the phyllospheric microbial communities over the course of a spatiotemporal varying environment with the development of tobacco brown spots, highlighting the importance of community succession.

Distinct Responses of Biofilm Carbon Metabolism to Nanoplastics with Different Surface Modifications.

Recently, there is an increasing concern regarding the toxicity of nanoplastics (NPs) on freshwater organisms. However, knowledge about the potential impacts of NPs with different surface modification on freshwater biofilms is still very limited. In this research, biofilms were cultured in lab and exposed to nano polystyrene (PS) beads: non-functionalized PS NPs, PS-COOH NPs, and the carbon source utilization of biofilms were measured by BIOLOG ECO microplates. The results showed that both two types of PS NPs significantly reduced the total carbon metabolic activity of biofilms, compared with the controls, whereas the carbon metabolic rate increased notably, especially for the PS-COOH NPs treatments at day 14. Moreover, results from six categories of carbon sources analysis suggested that PS NPs with different surface chemical properties exhibit distinct effects on the carbon utilization of biofilms, and the divergent changes of the specific carbon source category were observed at day 21 from the two PS NPs treatments. In addition, the metabolic functional diversity of biofilms were not altered by the PS NPs treatments. These findings highlighted that chemical properties of NPs play an important role in the toxic effects on the carbon metabolism activities of the biofilms. This study offers new insights that nanoplastics of different chemical characteristics have the ability to affect the microbial-mediated carbon cycling process in aquatic ecosystems.

Effects of different organic fertilisers on the microbial functional diversity and bacterial communities in a tobacco soil

ABSTRACT A long-term field experiment was conducted to investigate the effects of organic manure (commercial organic manure, CFM1; cow manure, CFM2; green manure, CFM3) and chemical fertiliser (CK) on the microbial diversity of a tobacco-planted soil using Biolog ECO and 16S rRNA sequencing technology. The results showed that organic manure treatments increased the available nutrient content in the soil, including nitrogen (AN, 2.1–13.3%), potassium (AK, 14.2–31.8%), labile organic carbon (19.9–37.4%), and microbial biomass carbon (11.9–21.0%), as well as the mean weight diameter and geometric mean diameter. The average well colour development value and McIntosh diversity index in CFM1 treatment increased by 23.6% and 18.9%, respectively, compared with CK. The 16S rRNA sequencing analysis suggested that CFM1 and CFM2 treatments improved the richness and diversity of soil bacteria and recruited more potentially beneficial bacteria. Redundancy analysis showed that soil physicochemical properties, excluding AN and Olsen–P were positively correlated with the abundance of bacteria dominant in the CFM1 treatment. This indicates that application of CFM1 is a sound fertilisation strategy to improve soil fertility and create a good ecological environment.