Assessment of the Parameters Influencing Microbial Calcite Precipitation in Injection Experiments Using Taguchi Methodology

Soil improvement is one of the major concerns in civil engineering. Therefore, a variety of approaches have been employed for different soil types. The loose granular soils and sediments have always imposed challenges due to their low strength and bearing capacity as well as presenting difficulties in drilling and excavation. Biomediated soil improvement, i.e., utilizing some bacteria to precipitate calcite on soil particles, has recently been introduced as a novel link of biotechnology and civil engineering to improve the problematic soils. Biogrout as a branch of biomediated soil improvement is based upon microbial calcium carbonate precipitation (MICP). In the present study, the Taguchi method with the aim of optimizing the process was utilized to design the experiments (DOE). A standard L9 orthogonal array with four parameters comprising bacterial cell concentration, molar concentration ratio of nutrient solution, curing time, and flow rate, each assigned to three levels, was selected. In this regard, soil samples were stabilized in sandy soil columns. Two-phase injections were conducted by injecting the bacterium Sporosarcina pasteurii PTCC 1642 in the first phase and nutrient in the second phase. Specimens were subjected to an unconfined compressive strength (UCS) test. ANOVA pointed out how effectual each parameter was. The most effective parameter was curing time, which accounted for 45.97% of the overall variance of the experimental data followed by bacterial cell concentration (22.01%), nutrient strength (19.98%), and flow rate (12.04%). Predicted UCS values for the optimum condition were validated in a confirmation test. Indeed, the UCS of the soil increased from 85 kPa in the control sample to 930 kPa for the optimally treated specimen. It was concluded that rather than curing time, the other parameters are almost equally influential in the applied injection procedure.     

Optimization of Influencing Parameters on Phenanthrene Removal Efficiency in Soil Washing Process by Using Response Surface Methodology

Response surface methodology (RSM) under Box–Behnken design (BBD) was applied to evaluate the effect of the influencing parameters including surfactant concentration, liquid/soil ratio, Humic Acid concentration, and washing time on phenanthrene removal efficiency in soil washing process by using the nonionic surfactant Tween 80 and find an optimal operational conditions to achieve the highest removal efficiency. A polynomial quadratic model was used to correlate phenanthrene removal efficiency and four independent variables (R² = 0.9719). Based on the obtained results the most influential parameter on phenanthrene removal efficiency was surfactant concentration with an impact value of 69.519%. Liquid/soil ratio was also another factor that significantly influenced on removal efficiency with an impact value of 25.014%. The interaction between surfactant concentration and liquid/soil ratio was also shown to have a positive significant effect on removal efficiency (p_value = 0.0027). However, the other independent variables Humic Acid concentration and time were not significant in the ranges selected in this study. Based on the optimization results maximum removal efficiency of 70.692 ± 3.647% was achieved under the conditions of surfactant concentration 5000 mg L^?1, liquid/soil ratio 30 v/w, HA concentration 9.88 mg L^?1, and washing time 2 h, which was in good agreement with predicted value (66.643%).     

Effects of antibiotics in soil on the population dynamics of transposon Tn5 carrying Pseudomonas fluorescens

The effects of kanamycin and streptomycin added to soil on the survival of transposon Tn5 modified Pseudomonas fluorescens strain R2f were investigated. Kanamycin in high (180 g g^-1 dry soil) or low (18 g g^-1) concentration or streptomycin in low concentration in Ede loamy sand soil had no noticeable effect on inoculant population dynamics in soil and wheat rhizosphere, whereas streptomycin in high concentration had a consistent significant stimulatory effect, in particular in the wheat rhizosphere. Streptomycin exerted its effect by selecting P. fluorescens with Tn5 insertion whilst suppressing the unmodified sensitive parent strain, as evidenced by comparing the behaviour of these two strains in separate and mixed inoculation studies.Soil textural type influenced the effect of streptomycin on the Tn5 carrying inoculant; the effect was consistently detected in rhizosphere and rhizoplane samples of wheat grown in Ede loamy sand after 7 and 14 days incubation, whereas it was only apparent after 7 days in rhizoplane or rhizosphere (and bulk soil) samples of wheat grown in two silt loam soils. Modification of soil pH by the addition of CaCO₃ or bentonite clay resulted in an enhancement of the selective effect of streptomycin by CaCO₃ and its abolishment by bentonite clay.The addition to soil of malic acid or wheat root exudate, but not of glucose, enhanced the streptomycin selective effect on the Tn5-modified P. fluorescens strain. Neither the streptomycin producer Streptomyces griseus nor two non-inhibiting mutants obtained following UV irradiation affected the dynamics of P. fluorescens (chr::Tn5) in soil and wheat rhizosphere.The effect of streptomycin in soil on inoculant Tn5 carrying bacteria depends on conditions such as soil type, the presence of (wheat) root exudates and the type of available substrate.     

Environmental Factors Affecting the Compressive Strength of Microbiologically Induced Calcite Precipitation-Treated Soil

Some microorganisms such as Sporoscarcina pasteurii precipitate calcium carbonate and are suitable for biocementation. This study aimed to investigate the effects of several factors including concentration of bacteria, chemical reactants, temperature, and pH on precipitation of calcium carbonate. The results showed that after 7 and 14 days of curing, the compressive strength of silty clay soil samples increased steadily as pH increased from 5 to 9. It was observed that pH plays an important role in biocementation. The highest compressive strength (i.e. 92 kPa) was observed when the soil was treated with 50 ml of bacterial solution after 14 days of curing. In addition, it was observed that the highest compressive strength of samples was achieved when the temperature was 40°C.     

Over-expression of gsh1 in the cytosol affects the photosynthetic apparatus and improves the performance of transgenic poplars on heavy metal-contaminated soil

Recent studies of transgenic poplars over‐expressing the genes gsh1 and gsh2 encoding γ‐glutamylcysteine synthetase (γ‐ECS) and glutathione synthetase, respectively, provided detailed information on regulation of GSH synthesis, enzymes activities and mRNA expression. In this experiment, we studied quantitative parameters of leaves, assimilating tissues, cells and chloroplasts, mesophyll resistance for CO₂ diffusion, chlorophyll and carbohydrate content in wild‐type poplar and transgenic plants over‐expressing gsh1 in the cytosol after 3 years of growth in relatively clean (control) or heavy metal‐contaminated soil in the field. Over‐expression of gsh1 in the cytosol led to a twofold increase of intrafoliar GSH concentration and influenced the photosynthetic apparatus at different levels of organisation, i.e., leaves, photosynthetic cells and chloroplasts. At the control site, transgenic poplars had a twofold smaller total leaf area per plant and a 1.6‐fold leaf area per leaf compared to wild‐type controls. Annual aboveground biomass gain was reduced by 50% in the transgenic plants. The reduction of leaf area of the transformants was accompanied by a significant decline in total cell number per leaf, indicating suppression of cell division. Over‐expression of γ‐ECS in the cytosol also caused changes in mesophyll structure, i.e., a 20% decrease in cell and chloroplast number per leaf area, but also an enhanced volume share of chloroplasts and intercellular airspaces in the leaves. Transgenic and wild poplars did not exhibit differences in chlorophyll and carotenoid content of leaves, but transformants had 1.3‐fold fewer soluble carbohydrates. Cultivation on contaminated soil caused a reduction of palisade cell volume and chloroplast number, both per cell and leaf area, in wild‐type plants but not in transformants. Biomass accumulation of wild‐type poplars decreased in contaminated soil by more than 30‐fold, whereas transformants showed a twofold decrease compared to the control site. Thus, poplars over‐expressing γ‐ECS in the cytosol were more tolerant to heavy metal stress under field conditions than wild‐type plants according to the parameters analysed. Correlation analysis revealed strong dependence of cell number per leaf area unit, chloroplast parameters and mesophyll resistance with the GSH level in poplar leaves.     

Effect of Water Stress Induced by Polyethylene Glycol on Growth,Proline Accumulation in <i>Agave americana</i> L.

The effect of water deficit was determined on both in vitro and soil seedling as well as in cells in suspension of Agave americana L. In order to do the establishment of cells, the formation of callus was induced; for it two auxins were evaluated: 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-mino-3,5,6-trichloropicolinic acid (picloram) at three concentrations (0.25, 0.5 and 0.75 mg L⁻¹) in three explants (leaf, root and meristems) cultured in MS semisolid medium. The callogenesis response was related to the type and section of the explant, as well as the regulator used, and a cell suspension was established using 0.5 mg L⁻¹ naphthaleneacetic acid (NAA) + 0.5 mg L⁻¹ Benzylaminopurine (BAP). Seedlings were exposed to polyethyleneglycol (15% and 30% w/v) with a water potential of −0.87 and −2.67 MPa, respectively, under soil conditions. Water stress was applied through restricted irrigation. Fresh weight, root system growth, and chlorophyll concentration were some of the parameters that were affected by the effect of water deficit on A. americana L. Chlorophyll concentration values were significantly decreased by 15 at 30% PEG (19.6 SPAD units) compared to the control treatment. In in vitro plants, the highest concentration of proline was found in the roots, being the treatment with 30% polyethylene glycol where the highest concentration of this osmoregulator was obtained (62.5 mg g⁻¹ DW). Under restricted irrigation conditions, an increase in proline concentration was observed both in the aerial part (2.2 µg 100 g⁻¹ DW) and in the root system (1.8 µg 100 g⁻¹ DW). However, the concentrations found were approximately ten times greater, less than those found under in vitro conditions. Therefore, the accumulation of proline can be considered an indicator of stress in Agave Americana L. growth in vitro.     

In Situ emulsification and mobilization of gasoline range hydrocarbons using surfactants

In this article the conditions that govern surfactant‐enhanced emulsification and mobilization of petroleum hydrocarbons in soil are reviewed. The effect of soil properties, groundwater constituents, and differing surfactant solutions on the emulsification process is discussed. A constant head soil flushing apparatus used to characterize surfactant‐enhanced mobilization of m‐xylene is described. Data showing the effect of surfactant‐enhanced mobilization on m‐xylene removal efficiency in washed sand is presented. Flushing solutions were used at concentrations from below to well above the critical micelle concentration (CMC) of the surfactants used. Removal efficiencies are shown to vary with surfactant concentration and with surfactant type. Flushing solutions of anionic, nonionic, and anionic/nonionic surfactant mixtures were evaluated.     

Cultivation, mineral nutrition and seed production of Catharanthus roseus (L.) G. Don in the temperate climate zone

The following aspects of Catharanthus roseus (L.) G. Don cultivation are covered in this chapter: climatic and soil requirements, agricultural measures, including propagation, sowing, planting, fertilization, irrigation and harvest date. The influence of these parameters on quality of herb material and alkaloid concentration is also discussed. Although tropical and subtropical zone have been reported as the most suitable for the cultivation of Catharanthus roseus, it seems to be possible to grow it also under other climatic conditions. However then, adequate conditions should be provided, according to the specific requirements of this species. Proper air temperature and humidity as well as sufficient soil moisture during growth, development and seed set are important factors for growing of Catharanthus roseus in the temperate zone. Albeit it may tolerate unfavourable environment conditions such as drought or salinity, its yield may be reduced considerably. Special attention has been paid in this paper to the effect of different ways of fertilization on alkaloid yield.     

In vitro development of maize immature embryos: a tool for embryogenesis analysis

During monocot embryo development, the zygote goes through a proembryo stage characterized by a radial symmetry and later becomes a true embryo with a bilateral symmetry. In order to determine culture conditions for immature embryonic stages, proembryos and embryos were isolated from controlled pollinated maize plants and developed in vitro. Precise culture conditions were determined for each type of explant: a monolayer system for embryos using NBM medium enriched with maltose (0.25 M) but without hormones, and a bilayer system for proembryo stages using N6 medium supplemented with maltose (0.35 M) and zeatin (3 mM). Morphological, cytological, and in situ hybridization analysis have shown that the resulting embryos (stages 1-2), developed in vitro, were similar to those formed in vivo and subsequently gave rise to fertile plants. This work demonstrates that successful embryo differentiation is dependent on specific parameters including the genotype, the nature of the carbon source, the type and concentration of hormones used and orientation of the embryos on the medium. The potential use of these results for embryo rescue and mutant analysis are discussed.     

CARAGANA FABR. PROMOTES REVEGETATION AND SOIL REHABILITATION IN SALINE-ALKALI WASTELAND

To determine how plantations of Caragana microphylla shrubs affect saline-alkali soil amelioration and revegetation, we investigated the vegetation and sampled soils from saline-alkali wasteland (SAW), perennial Caragana forestland (PCF), Caragana forest after fire disturbance (CFF). Results showed that with the development of Caragana Fabr., highly dominant species of Poaceae family, including Elymus dahuricus, Thermopsis lanceolata, Stipa tianschanica, died out in PCF. Moreover, Papilionaceae family, including Lespedeza indica, Oxytropis psammocharis, and Astragalus scaberrimus, was established both in PCF and CFF. Phytoremediation of saline-alkali wasteland (SAW) was achieved by plantation, resulting in the reduced soil pH, sodium adsorption ratio, exchangeable sodium percentage, salinity, and Na⁺ concentration around Caragana shrubs. Greater amounts of soil organic, total nitrogen, ammonium nitrogen, available phosphorus, and available potassium were observed in PCF topsoil than in SAW topsoil. The concentration of mineralized N in PCF soil was significantly lower than that in SAW soil at all sampled depths, indicating that Caragana shrubs were just using N and therefore less measured in soils. Fire disturbance resulted in decreased soil pH and salinity, but increased organic content, total nitrogen, and ammonium nitrogen. The improved soil parameters and self-recovery of shrubs indicated that Caragana shrubs were well established after burning event.     

青藏高原不同草地利用方式对土壤粒径分形特征的影响

研究青藏高原草地土壤粒径结构分形特征,为该地区土壤质量评价和生态恢复提供科学依据。以青藏高原4种高寒草地(放牧、围栏禁牧、围栏禁牧+补植、未干扰)为对象,采用分形理论,研究不同利用方式对高寒草地土壤颗粒组成及分形特征的影响,明确土壤粒径分形特征的影响因素。结果表明：与放牧和围栏禁牧+补植相比,围栏禁牧草地中黏粒和粉粒体积分数分别增加了60%—91.1%、43.5%—80.1%,禁牧能够促进土壤砂粒向黏粒和粉粒转变。不同草地利用方式对分形维数有显著影响,单重分形维数D值依次为放牧草地<围栏禁牧+补植草地<未干扰草地=围栏禁牧草地,多重分形维数,包括信息维数D₁、信息维数/容量维数比值D₁/D₀和关联维数D₂依次为放牧草地<围栏禁牧+补植草地<围栏禁牧草地<未干扰草地。单重分形维数D与土壤黏粒、粉粒呈极显著正相关(P<0.01);砂粒、黏粒、粉粒、有机碳和全氮是多重分形维数的限制因素。信息维数D₁、信息维数/容量维数比值D₁...     

小果核果茶扦插繁殖技术研究

小果核果茶为山茶科石笔木属常绿乔木,树形美观,叶泽光亮,洁白亮丽的花朵和茂密的果实具有较高的观赏价值;而且其种子含油量高,是一种具有潜在开发价值的新型生物能源植物。该研究采用4因素3水平的正交设计,分析了激素种类、处理浓度、浸泡时间和扦插基质对小果核果茶扦插生根的影响,并以生根率、生根数和平均根长为扦插生根效果的评价指标,构建生根效果指数,总体评价小果核果茶的生根效果。结果表明:对生根效果指数影响最大的是扦插基质,激素种类次之,影响最小的是处理时间。其中,扦插基质对生根效果指数的影响达到显著水平(P0.05);当激素种类为NAA、处理浓度为500mg·L~(-1)、处理时间为12 h、扦插基质为泥炭土时,小果核果茶扦插的生根效果最好。该研究结果为小果核果茶的栽培生产、育种以及资源开发和利用提供了科学依据。     

Phytoremediation of soil contaminated with PCBs using different plants and their associated microbial communities

In this work, we evaluate the abilities of the plants Brassica juncea, Avena sativa, Brachiaria decumbens, and Medicago sativa to uptake polychlorinated biphenyls (PCBs) and induce degradation of soil microorganisms from contaminated soil. Removal of PCBs 44, 66, 118, 153, 170, and 180 was evaluated in both rhizospheric and nonrhizospheric soils. Microbial and bphA1 gene quantifications were performed by real-time PCR. The PCB concentrations in plant tissues and soil were determined, and a fluorescein diacetate (FDA) hydrolysis assay was used to measure microbial activity in soil. The removal percentages for all PCB congeners in planted soil versus unplanted control soil were statistically significant and varied between 45% and 63%. PCBs 118, 153, 138, and 170 were detected in Brachiaria decumbens roots at different concentrations. In planted soil, an increase in the concentration of bacteria was observed compared to the initial concentration and the concentration in unplanted control soil; however, no significant differences were identified between plants. The number of copies of the bphA1 gene was higher in rhizospheric versus non- rhizospheric soil for all plants at the end of the experiment. However, alfalfa and oat rhizospheric soil showed significant differences in the copy number of the bphA1 gene. In general, the concentration of fluorescein in the rhizospheric soil was greater than that in the nonrhizospheric soil. Although the plants had a positive effect on PCB removal, this effect varied depending on the type of PCB, the plant, and the soil.     

Cadmium Uptake by Lettuce (Lactuca sativa L.) as Basis for Derivation of Risk Limits in Soils

The availability and uptake of Cd by lettuce (Lactuca sativa L.) in two common tropical soils (before and after liming) were studied in order to derive human health-based risk soil concentration. Cadmium concentrations ranging from 1 to 12 mg kg^?1 were added to samples from a clayey Oxisol and a sandy-loam Ultisol under glasshouse conditions. After incubation, a soil sample was taken from each pot, the concentration of Cd in the soil was determined, lettuce was grown during 36 d, and the edible parts were harvested and analyzed for Cd. A positive linear correlation was observed between total soil Cd and the Cd concentration in lettuce. The amount of Cd absorbed by lettuce grown in the Ultisol was about twice the amount absorbed in the Oxisol. Liming increased the soil pH and slightly reduced Cd availability and uptake. CaCl₂ extraction was better than DTPA to reflect differences in binding strength of Cd between limed and unlimed soils. Risk Cd concentrations in the Ultisol were lower than in the Oxisol, reflecting the greater degree of uptake from the Ultisol. The derived risk Cd values were dependent on soil type and the exposure scenario.     

Enhanced inhibition of Aspergillus niger on sedge (Lepironia articulata) treated with heat‐cured lime oil

Aims

This study aimed to examine heat curing effect (30–100°C) on antifungal activities of lime oil and its components (limonene, p‐cymene, β‐pinene and α‐pinene) at concentrations ranging from 100 to 300 μl ml^?1 against Aspergillus niger in microbiological medium and to optimize heat curing of lime oil for efficient mould control on sedge (Lepironia articulata).

Methods and Results

Broth dilution method was employed to determine lime oil minimum inhibitory concentration, which was at 90 μl ml^?1 with heat curing at 70°C. Limonene, a main component of lime oil, was an agent responsible for temperature dependencies of lime oil activities observed. Response surface methodology was used to construct the mathematical model describing a time period of zero mould growth on sedge as functions of heat curing temperature and lime oil concentration. Heat curing of 90 μl ml^?1 lime oil at 70°C extended a period of zero mould growth on sedge to 18 weeks under moist conditions.

Conclusions

Heat curing at 70°C best enhanced antifungal activity of lime oil against A. niger both in medium and on sedge.

Significance and Impact of the Study

Heat curing of lime oil has potential to be used to enhance the antifungal safety of sedge products.     

Growth promotion of groundnut by IAA producing rhizobacteria Bacillus licheniformis MML2501

B. licheniformis MML2501 which was isolated from groundnut rhizosphere soil showed increased populations on spermozphere colonisation and significantly increased the seed germination and other growth parameters in groundnut under in vitro conditions. B. licheniformis MML2501 did not solubilise phosphate but produced indole acetic acid (IAA), with a maximum of 23 μg/ml under optimised conditions such as pH 7.0, temperature 35°C, tryphtophan at a concentration of 16 mM and at 200 rpm shaken conditions. The production of IAA by B. licheniformis MML2501 was further confirmed by TLC and HPLC analyses, in which the R_f value and retention time of 0.66 and 4.1 min respectively, match with that of the authentic IAA. Seed treatment of B. licheniformis MML2501 in groundnut showed a significant increase in seed germination, other growth parameters and yield parameters under potted plant experiments.     

黄土丘陵区植被类型与土壤微生物区系及生物量的关系

以陕西延安黄土丘陵区5种不同植被类型(人工刺槐林、天然侧柏林、天然辽东栎林、灌丛和裸地)为研究对象,分析了土壤微生物生物量碳、氮含量、细菌和真菌的丰度变化规律及其与土壤基本化学性质的关系。结果表明:(1)4种植被类型的土壤质量较之裸地都有不同程度的改善,总体趋势:天然林地人工林地裸地;(2)土壤微生物生物量碳、氮的总体趋势:天然林地最高,人工林次之,裸地最低,且与土壤有机碳(SOC)、全氮(TN)和速效磷(AP)极显著正相关(P0.01);(3)裸地土壤的细菌丰度最低,人工刺槐林真菌含量显著低于天然辽东栎林。细菌丰度与土壤营养状况呈显著正相关(P0.05),而真菌与土壤营养无明显相关性,只与土壤pH负相关。说明在该研究区域,植被类型与土壤质量对微生物资源都具有不同程度的作用。     

Carbon and nitrogen status of litterfall, litter decomposition and soil in even-aged larch, red pine and rigitaeda pine plantations

The carbon (C) and nitrogen (N) status in forest ecosystems can change upon establishment of plantations because different tree species have different nutrient cycling mechanisms. This study was carried out to evaluate C and N status of litterfall, litter decomposition and soil in three adjacent plantations consisting of one deciduous (larch: Larix leptolepis) and two evergreen (red pine: Pinus densiflora; rigitaeda pine: P. rigida × P. taeda) species planted in the same year (1963). Both the pine plantations showed comparatively higher C input from needle litter but significantly lower N concentration and input than the larch plantation (P < 0.05). During the decomposition process, the deciduous larch needle litter showed low C concentration and C remaining in soil, but high N concentration and N remaining in soil compared to the two evergreen pine needle litters. However, the soil C and N concentration and their content at a soil depth of 0–10 cm were not affected significantly (P > 0.05) by the plantation type. These results demonstrate the existence of considerable variation in C and N status resulting from needle litter input and litter decomposition in these three plantations grown at sites with similar environmental conditions.     

Phytoremediation-biorefinery tandem for effective clean-up of metal contaminated soil and biomass valorisation

During the last few decades, phytoremediation process has attracted much attention because of the growing concerns about the deteriorating quality of soil caused by anthropogenic activities. Here, a tandem phytoremediation/biorefinery process was proposed as a way to turn phytoremediation into a viable commercial method by producing valuable chemicals in addition to cleaned soil. Two agricultural plants (Sinapis alba and Helianthus annuus) were grown in moderately contaminated soil with ca. 100 ppm of Ni and further degraded by a fungal lignin degrader—Phanerochaete chrysosporium. Several parameters have been studied, including the viability of plants, biomass yield, and their accumulating and remediating potentials. Further, downstream processing showed that up to 80% of Ni can be easily extracted from contaminated biomass by aqueous extraction at mild conditions. Finally, it was demonstrated that the growth of plants on the contaminated soil could be degraded by P. chrysosporium, and the effect of nickel and biomass pretreatment on the solid-state fermentation was studied. The proposed and studied methodology in this work could pave the way for successful commercialization of the phytoremediation process in the near future.