Study on Effect of Nitrogen and Phosphorous on the Resistance of E.coli to Chloramphenicol and its Mechanism

This study was designed to explore the effect of nitrogen and phosphorous on the resistance of E.coli from environment and the mechanism.Microcosms were established to study the effect of nitrogen and phosphorous on the resistant phenotype of E.coli to chloramphenicol (CHL).cat gene of isolated drug-resistant strains and susceptible strains were detected.The results showed that,different concentration of nitrogen and phosphorous could induce the formation of antibiotics resistance of E.coli to CHL.The rate of cat gene of 46 strains of chloramphenicol resistant E.coli was 89.13%,which was 0 in the 16 strains of chloramphenicol sensitive E.coli.The results indicated that,nitrogen and phosphorous in the microcosms could induce the formation and maintenance of resistance to chloramphenicol in E.coli,which had correlation with cat gene.     

Increase in the Nitrogen Content of Soil by the Introduction of Earthworms into Soil

The contribution of an earthworm species ( Amynthas vittatus ) to the increase of the nitrogen content of soil was examined. Three specimens of adult earthworms were introduced into 300 g of soil (Gray Lowland soil, silty clay) supplemented with 1% carboxymethyl cellulose in a container and incubated for 32 d at 22°C in the dark. The contents of total-N, NH₄-N and NO₃-N, and the population of aerobic nitrogen-fixing bacteria in soil significantly increased after incubation with the earthworms, while the natural abundance of ¹⁵N (δ¹⁵N) in soil decreased. The amount of nitrogen in the earthworms did not decrease during the incubation in the microcosm. Both acetylene reduction activity of the microcosm and incorporation of ¹⁵N to soil from atmospheric ¹⁵N₂ were significantly enhanced by the introduction of the earthworms into soil, though the observed increment of nitrogen in soil was much higher than the estimated one based on the nitrogen-fixing activity. The results obtained in the present study indicated that the earthworms increased the nitrogen content of soil, presumably due to the enhancement of the nitrogen-fixing activity of the soil from the microcosm by the earthworms.     

Effect of potassium phosphate fertilization on production and emission of methane and its C-stable isotope composition in rice microcosms

Rice fields are an important source for atmospheric CH₄, but the effects of fertilization are not well known. We studied the turnover of CH₄ in rice soil microcosms without and with addition of potassium phosphate. Height and tiller number of rice plants were higher in the fertilized than in the unfertilized microcosms. Emission rates of CH₄ were also higher, but porewater concentrations of CH₄ were lower. The δ¹³C values of the emitted CH₄ and of the CH₄ in the porewater were both 2-6% higher in the fertilized microcosms than in the control. Potassium phosphate did not affect rate and isotopic signature of CH₄ production in anoxic soil slurries. On the other hand, roots retrieved from fertilized microcosms at the end of incubation exhibited slightly higher CH₄ production rates and slightly higher CH₄-δ¹³C values compared to roots from unfertilized plants. Addition of potassium phosphate to excised rice roots generally inhibited CH₄ production and resulted in increasingly lower δ¹³C values of the produced CH₄. Fractionation of ¹³C during plant ventilation (i.e. δ¹³C in pore water CH₄ versus CH₄ emitted) was larger in the fertilized microcosms than in the control. Besides plant ventilation, this difference may also have been caused by CH₄ oxidation in the rhizosphere. However, calculation from the isotopic data showed that less than 27% of the produced CH₄ was oxidized. Collectively, our results indicate that potassium phosphate fertilization stimulated CH₄ emission by enhancing root methanogenesis, plant ventilation and/or CH₄ oxidation, resulting in residence times of CH₄ in the porewater in the order of hours.     

Vertical advection of extracellular DNA by water capillarity in soil columns

The fate of extracellular DNA in the environment concerns both the fate of transgenes from genetically modified organisms and the evolution of active bacteria capable of incorporating this DNA into their genomes. This study addressed the possibility that DNA, like other organic molecules, could move vertically in the capillary fringe of groundwater aquifers. The targeted gene fragment used here was the 35S-nptII sequence, which was below detection levels in controls. Initial microcosm studies detected the DNA target molecule by PCR during the entire experiment. The vertical advection of water and DNA were monitored for a period of 3 days in soil columns. DNA was added as a water solution at the bottom of the unsaturated soil column, and then DNA-free water was added at the bottom after 12 and 24 h. After the addition of the DNA solution, capillary water rose 4 cm within the soil column and the target DNA was detected up to that height. After 60 min, the entire soil column (10 cm) was wetted and the target sequence was detected up to a height of 7.5 cm. After the second wetting (12 h later), the target sequence was detected up to the top of the soil column (10 cm). However, after the third wetting (24 h later), the marker sequence was only found at heights from 0.5 to 4 cm. Results clearly show the vertical movement of DNA due the capillary rise and suggest the possibility of DNA degradation within the soil column.     

Effects of rhamnolipids on bacterial communities in a dioxin-contaminated soil and the gut of earthworms added to the soil

The biosurfactants rhamnolipids and the “soil ecosystem engineers” earthworms are often used to remediate contaminated soils. However, the effects of rhamnolipids on earthworm intestinal flora and microbial community in soil containing earthworms are not clearly understood. In our study, a 21-d microcosm experiment was carried out to reveal the effects of rhamnolipids on microbial abundance, composition, and metabolism, as well as contaminant degradation capacity. Both rhamnolipids and earthworm...     

Effects of weathering state of coarse‐soil fragments on tree‐seedling nutrient uptake

Fine earth accumulated within the weathering fissures of the coarse‐soil fraction (particles > 2 mm), so called “stone‐protected fine earth”, can provide a high short‐term nutrient release by cation exchange. It is thus hypothesized that unweathered gneiss particles cannot provide plants with exchangeable‐cation nutrients and that biological weathering is needed to include silicate‐bound nutrients into biochemical cycles. In a microcosm experiment, ectomycorrhizal Norway spruce (Picea abies) seedlings were grown on either weathered or unweathered paragneiss coarse‐soil fragments under natural hydraulic and climatic boundary conditions. A nutrient solution containing N, P, and K was added, however Mg and Ca could only be taken up from the coarse‐soil substrate. Solutes in drainage were analyzed during the experiment; plant nutrient uptake was determined after the experiment ended. Solute dynamics depended on the weathering state of the substrates: unweathered gneiss showed high initial Mg and Ca fluxes that diminished strongly afterwards, whereas weathered gneiss showed a much more gradual and sustainable release of these cations. Patterns in dissolved organic C and sulfate drainage indicated that the internal pores of weathered gneiss fragments contained organic material most likely as a result of living spaces from microorganisms. Plant biomass did not differ between treatments, however Mg content was higher in seedlings grown on weathered gneiss. Nutrient budgets demonstrated that the “stonesphere” of weathered gneiss can act as a quasi‐constant nutrient source whereas unweathered gneiss only provided high initial nutrients fluxes. In nutrient‐depleted, acidified fine‐earth environments, the coarse‐soil fraction may therefore act as a retreat for nutrient‐adsorbing tissues and as a buffer for nutrient shortages.     

植穴控制体对盐渍土水分运移的影响

该研究旨在探讨以植穴控制体作为调控水分运移的基本单元,以提高盐渍土造林成效及土地使用价值。结合土柱微宇宙试验控制技术,选取植穴控制体（PHCB）的结构因子（木本植物、植穴袋材料、覆膜材料配比、填充材料配比及填料厚度）和环境因子（地下水矿化度、土壤质地、补水量）为影响因素,设计U12(12×43×34)的混合均匀试验。经回归分析筛选出PHCB最佳结构设计参数,建立植穴控制体结构因子和环境因子与含水率间关系的预测方程。分析结果显示：当每次补水量达到5.06 mm,土壤质地为轻黏土的环境条件下,木本植物选择紫叶李、植穴袋材料为麻布、PVA（polyvinyl alcohol）/VAE（vinyl acetate-ethylene copolymer emulsion）/蒙脱石的体积比按100︰20︰10配制成覆膜材料,秸秆、枯枝落叶和木屑以质量比1︰1︰2混合制成填料,可使植穴袋内的土壤含水率提高48.83%。而环境因子的地下水矿化度及结构因子填料厚度对植穴内水分运移的影响均不显著。PHCB可以有效调控盐渍土水分运移过程,是一种盐渍土生态修复方法。     

Effects of collembola on nutrient cycling and microbial biomass on intact soil microcosm

(Jpn. J. Soil Sci.Plant Nutr., 77, 299–306, 2006)

The effects of Collembola (Folsomia candida Willem) on nutrient cycling, microbial biomass, and soil respiration were studied using intact soil microcosms. Intact soil microcosms (dia. 10·6 cm and depth 15 cm) were taken from pine forest soil, and were divided into four treatments · the unmanipulated control and three Collembolan manipulations in which microcosms were defaunated by deep-freezing, and then F. candida were introduced at three densities (0, 50, 100 per microcosm). The microcosms were incubated on forest floor with a roof. At 3- to 4-week intervals the microcosms were irrigated with deionized water for analyses of nutrients (Na+, K+, NH4+, Ca2+, Mg2+, Cl?, NO3?, SO42?) in the leachate. Soil respiration was measured using an infrared gas analyser. After 13 and 34 weeks of exposure, microcosms were destructively sampled. Collembola did not significantly affect microbial biomass C, N, and P nor soil respiration. Because the experiment was started in winter, nutrient leaching increased from spring to summer with increasing microbial activity. At the end of the experiment, leached nitrate from microcosms was significantly different between the 0 and 50 Collembolan treatments. Total established Collembolan biomass was under 4% of the soil microbial biomass in the microcosms, while manipulation of Collembola affected soil nitrogen dynamics at high microbial and collembolan activity.     

Response of soil enzymes to Linear Alkylbenzene Sulfonate (LAS) addition in soil microcosms

Soil enzymatic activities (phosphatases, arylsulphatase and dehydrogenase) were measured in microcosm systems designed for the study of the impact of a commercial mixture of Linear Alkylbenzene Sulphonate (LAS) homologues on a xerofluvent agricultural soil. The soil microcosms consisted of glass columns filled with 800 g of dry soil which were fed with sterile commercial LAS solutions at concentrations of 10 or 50 mg l⁻¹ for periods of time up to 21 days. A soil microcosm fed with sterile distilled water was included in this study and considered as control. Our results showed that the continuous application of the anionic surfactant to soil increased the values of the enzymes acid and alkaline phosphatases and arylsulphatase. On the contrary, the dehydrogenase activity was decreased by the continuous application of 10 or 50 mg l⁻¹ LAS when compared with control microcosms. In addition, a statistically negative correlation was found between this enzymatic activity in the upper portion of the soil columns amended with LAS and the viable counts of heterotrophic aerobic microorganisms. Moreover, in order to test the influence of LAS on nutrient availability and, consequently, on bacteria populations and soil biological activities, phosphate concentration was regularly determined in the microcosm leachates. The phosphate concentration tested in the leachate of the microcosm continuously amended with 50 mg l⁻¹ LAS solution was significantly lower than the concentrations detected in the leachate of the microcosms continuously amended with 10 mg l⁻¹ LAS throughout the experiment.     

氮磷营养盐对大肠杆菌四环素耐药性的影响

根据临床和实验室标准协会（clinical and laboratory standards institute, CLSI）推荐的微量肉汤稀释法检测泥样中大肠杆菌对四环素的敏感性,并应用PCR法检测四环素耐药基因tetA,tetB,探讨氮磷营养盐对大肠杆菌四环素耐药性影响的可能机制。结果表明：添加不同剂量氮磷营养盐能够导致大肠杆菌对四环素产生耐药性,但大肠杆菌的耐药率与剂量间无明显相关性。37株高耐药率大肠杆菌tetA基因的阳性率为100%,66株低耐药率大肠杆菌tetA基因的阳性率为16.67%,而16株对药物敏感的大肠杆菌未检测到tetA基因;但供试大肠杆菌均未检测到tetB基因。这表明,氮磷营养盐诱导的大肠杆菌四环素耐药性与tetA基因有关。