Ecotoxicological Evaluation of a Bench-Scale Bioslurry Treating Explosives-Spiked Soil

The ecotoxicological effects of four bioslurry reactors treating 2,4,6-trinitotoluene (TNT)- and 1,3,5-trinitro-1,3,5-triazacyclohexane (RDX)-spiked soil were evaluated. A control bioslurry reactor was used to assess the endogenous toxicity of the bioslurry operation conditions. A battery of ecotoxicity tests was used: Microtox, green algae growth inhibition, bacterial genotoxicity and mutagenicity, and earthworm mortality and growth inhibition. Bioslurry soluble and solid phases were separated by centrifugation in order to identify toxicity and possible toxicants associated with each phase. Microtox toxicity values were initially very high in both bioslurry reactors spiked with TNT, in relation with TNT concentration. Initial toxicity was also detected by algal growth inhibition, earthworm lethality, genotoxicity and mutagenicity tests. An endogenous toxicity was detected in the control bioreactor using the Microtox and the SOS Chromotest. The soluble phase of the control bioslurry was genotoxic, suggesting that some potentially genotoxic agents were induced in the bioslurry samples. At the end of the bioremediation treatment, data showed that toxicity was reduced using all of the bioassays, except for earthworm lethality and growth inhibition tests in both RDX-spiked bioslurries. This study demonstrates the usefulness of a battery of toxicity tests to monitor bioremediation processes.     

Stimulation of weed seed germination by 1–(3-chlorophthalirnido) cyclohexanecarboxamide (AC 94377)

The N-substituted phthalimide, AC 94377, promoted the germination of dry-stored seeds of 17 out of 24 weed species in Petri-dish tests maintained at 22°C. In a further test it was able to substitute for light and/or alternating temperatures in promoting the germination of some species. AC 94377 was more active than GA, in stimulating Solanum nigrum seed to germinate. When mixed with soil in the laboratory, AC 94377 again promoted germination of S. nigrum but the level and persistence of activity differed between soils. In the glasshouse, with soil containing a natural weed seed population, more seedlings emerged from treated than from untreated soil but this number represented fewer than half of the apparently viable seeds present. Field applications of AC 94377 promoted the germination of hand-sown seeds of certain weed species but did not increase the number of seedlings emerging from the natural weed seed bank.     

Toxicities of DDE on Wheat and Bioremediation of DDE by Fungus Pleurotus pulmonarius

An in vitro study was performed to determine the acute phytotoxicities and genotoxicity of DDE either spiked to soil or added to hydrophonic cultures on wheat Triticum aestivum. A 24-well plate was first used to determine toxicity on individual grains using conventional seed germination/seedling growth toxicity tests whereas a single cell electrophoresis system was applied to measure genotoxicity at single cell level for wheat. Hydrophonic cultures provide a simplified environment to screen for toxicities with high sensitivity. Inverse dose-response relationships were detected between exogenous DDE levels and one of the following parameters: seed germination, seedling growth, and genotoxicity. In contrast, soil reduced the stress on T. aestivum by lowering bioavailability leading to less DDE distributed in radicle and coleoptile, modulated growth, and enhanced tolerance. At all DDE doses spiked to soil including the reference safety level of 0.5 mg/kg, DNA breakage was detected in both radicle and coleoptile but their magnitudes did not correlate with the organ nor the soil DDE contents. Thus, although wheat is highly sensitive to the genotoxic effect of DDE, first demonstrated here, the seed germination test offers a simple quantitative measure of DDE's phytotoxicity in soil and hydrophonic cultures. This study also found that fungus Pleurotus pulmonarius, which secretes extracellular ligninolytic enzymes causing non-specific cleavage of lignin and organopollutants, remediated DDE spiked to soil. In 5 weeks, 78% of 10 mg/kg DDE was biodegraded, and the fungal-treated soil reduced acute toxicity on T. aestivum using the seed germination test.     

PFOS对蚯蚓急性毒性和回避行为的影响

全氟辛烷磺酰基化合物(PFOS)作为一种新型持久性有机污染物,已经成为环境科学和毒理学的研究热点,其对生态环境的影响值得深入研究.本文采用OECD标准滤纸接触法、人工土壤法及自然土壤法研究了PFOS对蚯蚓急性致死作用及回避行为的影响.结果表明： PFOS对蚯蚓的急性毒性作用与染毒时间和染毒浓度相关,试验求得滤纸法48 h、人工土壤法14 d和自然土壤法14 d的LC₅₀值分别为13.64 μg·cm^-2、955.28 mg·kg^-1和542.08 mg·kg^-1;人工土壤和自然土壤中蚯蚓在PFOS的最大试验浓度组160 mg·kg^-1中均表现出显著的回避行为,表明蚯蚓可以明显感知较高浓度PFOS污染土壤并作出回避反应.与急性毒性试验的测试终点LC₅₀相比,蚯蚓行为测试终点对PFOS的反应更为敏感.自然土壤中PFOS对蚯蚓的急性毒性大于人工土壤,相同浓度PFOS作用下,蚯蚓于自然土壤中的回避行为较人工土壤明显.     

甲胺磷、乙草胺和铜单一与复合污染对蚯蚓的毒性效应研究

以赤子爱胜蚓(Eisenia foetida)为例,通过滤纸急性毒性试验法研究了东北黑土区普遍存在的3种农用化学品甲胺磷、乙草胺和Cu对蚯蚓的单一与复合毒性效应．单一实验结果表明,三者对蚯蚓均有毒性,它们的毒性顺序为乙草胺＞甲胺磷＞Cu,半致死浓度分别为：0．307、0．708和118．70mg.kg^-1;同时通过观察可知,乙草胺与Cu以皮肤渗入蚯蚓体的毒性效应较甲胺磷明显．复合毒性实验结果表明,低浓度Cu与高浓度Cu对甲胺磷的毒性均有增强作用;低浓度Cu对乙草胺的毒性有削弱作用,但高浓度Cu对乙草胺的毒性有增强作用．可见,三者对土壤生态系统生态安全性和土壤健康质量存在潜在危害,同时这几种污染物的共存进一步加大了潜在危害性,且复合毒性效应与各组浓度组合密切相关．     

植冠与土壤种子库储存种子的萌发特性及策略

为明确黄土丘陵沟壑区植物种子库如何调控种子萌发来提高个体适合度,选择研究区7种具有种子库的主要物种为研究对象,以刚成熟和室内储存种子为对照,比较植冠宿存(5个宿存期)和土壤埋藏(5a埋藏期)对植物种子萌发特性的影响,探讨植冠种子库与土壤种子库储存下的种子萌发策略。结果表明:7种植物种子经过不同种子库储存后萌发特性表现出明显的种间差异,黄刺玫(Rosa xanthina)和水栒子(Cotoneaster multiflorus)种子萌发力表现为植冠宿存不变型、土壤储存增强型,土壤储存明显提高水栒子种子萌发速率;达乌里胡枝子(Lespedeza davurica)和狼牙刺(Sophora davidii)种子萌发力表现为植冠宿存增强型、土壤储存减弱型,种子萌发历时表现为植冠宿存延长型,土壤种子库储存还可加快达乌里胡枝子萌发速率、缩短萌发历时;茭蒿(Artemisia giralaii)和铁杆蒿(Artemisia gmelinii),种子萌发率随植冠宿存时间先升高后降低,随土壤储存时间先降低后升高,土壤储存可推迟其萌发,铁杆蒿种子萌发速率在植冠与土壤储存后均加快;紫丁香(Syringa oblata)种子萌发率随植冠宿存先升高后降低,土壤储存明显加快其种子启动萌发与速率。在黄土丘陵沟壑区,植物种子经过植冠或土壤种子库储存,或增加、加快、提早萌发充分利用有利条件提高占据性,或减少、减缓、推迟萌发分摊不利条件的风险;而且该区植物植冠与土壤储存后种子萌发特性间的关系,体现各自适应环境与应对干扰的分工与协作策略,主要表现为:单一主导型和相辅相成型。     

Enhanced Biodegradation of Petroleum Hydrocarbons in Contaminated Soil

Soil samples taken from a contaminated site in Northern Quebec, Canada, exhibited a low capacity for biodegradation of total petroleum hydrocarbons (TPH), despite a high capacity for the mineralization of aromatic hydrocarbons and a low toxicity of soil leachates as measured by Microtox assay. Toxicity assays directly performed on surface soil, including earthworm mortality and barley seedling emergence, indicated moderate to high levels of toxicity. Soil biostimulation did not improve the removal of petroleum hydrocarbons, while bioaugmentation of soil with a developed enrichment culture increased the efficiency of hydrocarbon removal from 20.4% to 49.2%. A considerable increase in the removal of TPH was obtained in a bioslurry process, enhancing the mass transfer of hydrocarbons from soil to the aqueous phase and increasing the efficiency of hydrocarbon removal to over 70% after 45 days of incubation. The addition of ionic or nonionic surfactants did not have a significant impact on biodegradation of hydrocarbons. The extent of hydrocarbon mineralization during the bioslurry process after 45 days of incubation ranged from 41.3% to 58.9%, indicating that 62.7% to 83.1% of the eliminated TPH were transformed into CO₂ and water.     

4种蚯蚓肠道微生物对砷毒性的响应差异研究

蚯蚓肠道是微生物多样性的一个潜在存储库。砷对蚯蚓肠道微生物群落的影响已被证实,但砷在不同蚯蚓肠道菌群中生物转化的差异仍不清楚。为了进一步阐述土壤中广泛存在的低浓度砷(浓度为5,15,25 mg/kg)对不同种类蚯蚓肠道微生物影响的差异,将4种典型蚯蚓暴露于砷污染土壤后,测定其肠道微生物组成变化,并分析砷对不同蚯蚓肠道内砷富集、形态和砷生物转化基因的影响。结果显示,所有蚯蚓组织内均存在明显的砷富集,其富集系数由高到低依次为:安德爱胜蚓(1.93)>加州腔蚓(0.80)>通俗腔蚓(0.78)>湖北远盲蚓(0.52),蚯蚓组织和肠道内砷形态主要以无机砷为主,其中As(III)含量比例> 80%,部分蚯蚓组织内还发现少量有机砷。4种蚯蚓肠道微生物群落在门水平上主要以变形菌、厚壁菌和放线菌为主,并与周围土壤细菌群落组成存在显著差异。同时,在土壤和肠道内共检测到17个砷转化基因,其中蚯蚓肠道内As(V)还原和砷转运相关基因相对丰度较高,而砷(去)甲基化基因丰度较低。此外,低浓度砷污染对蚯蚓生长无显著影响,却能引起蚯蚓肠道微生物群落的紊乱。蚯蚓种类和砷污染是引起蚯蚓肠道微生物...     

Phytoremediation of polycyclic hydrocarbon contaminated soil: part II. Impact on ecotoxicity

Several biological assays were used to evaluate the toxic effects of contaminants in soil after phytoremediation. During the treatment process, significant decreases in overall toxicity were observed. Specifically, earthworm survivability and lettuce germination increased over the study period. Microbial respiration improved, but only in planted treatments. Toxicity and total polycyclic aromatic hydrocarbon concentrations showed some correlation, but the relationships generally were not significant. Soil moisture was less of a predictor for biological responses. The presence of plants did not provide a clear advantage for improving toxicity compared to unplanted treatments.     

土壤种子库与矿业废弃地植被恢复研究I.Leonard瓶-罐装置在土壤种子库检测中的应用

１前言土壤种子库的组成和动态的检测主要有两种方法,其一是把土壤样品铺在垫有沙子（经消毒除去沙子内可能有的种子）的花盆或其它发芽框上,给予合适条件使土样中的种子萌发,记录幼苗数及种类［１～５］;另一种是用物理方法分离土样直接得到种子,检测种子活力和统计...     

Application of bioluminescence-based microbial biosensors to the ecotoxicity assessment of organotins

The toxicity of two common organotin pollutants and their initial breakdown products (tributyltin, dibutyltin, triphenyltin and diphenyltin) were assessed using two different bioluminescent microbial biosensors: Microtox and lux -modified Pseudomonas fluorescens pUCD 607. The organotins were made up as standards, and tested both in double-deionized water and in extracted soil solution, the latter representing a realistic matrix for terrestrial contamination. Microtox was especially sensitive to the organotins, with 50% effective concentration (EC₅₀) values (15 min) for tributyltin as low as 21·9 μg l⁻¹ in pure water, and 0·118 μg l⁻¹ in soil extract. The sensitivity of Microtox was increased by an order of magnitude in soil extract. The Ps. fluorescens was less sensitive, with EC₅₀ values (30 min) of around 800 μg l⁻¹ in pure water. The toxicity to Ps. fluorescens was decreased by around an order of magnitude in soil extract. The two biosensors showed different response patterns, with Microtox being more sensitive to the triorganotins and Ps. fluorescens being more sensitive to the diorganotins.     

Relative toxicity of arsenite and arsenate on germination and early seedling growth of rice (Oryza sativa L.)

Elevated soil arsenic levels resulting from long-term use of arsenic contaminated ground for irrigation in Bangladesh may inhibit seed germination and seedling establishment of rice, the country's main food crop. A germination study on rice seeds and a short-term toxicity experiment with different concentrations of arsenite and arsenate on rice seedlings were conducted. Percent germination over control decreased significantly with increasing concentrations of arsenite and arsenate. Arsenite was found to be more toxic than arsenate for rice seed germination. There were varietal differences among the test varieties in response to arsenite and arsenate exposure. The performance of the dry season variety Purbachi was the best among the varieties. Germination of Purbachi was not inhibited at all up to 4 mg l^–1 arsenite and 8 mg l^–1 arsenate treatment. Root tolerance index (RTI) and relative shoot height (RSH) for rice seedlings decreased with increasing concentrations of arsenite and arsenate. Reduction of RTI caused by arsenate was higher than that of arsenite. In general, dry season varieties have more tolerance to arsenite or arsenate than the wet season varieties.     

Phytotoxicity of Sodium Fluoride and Uptake of Fluoride in Willow Trees

The willow tree (Salix viminalis) toxicity test and a cress seed germination test (Lepidium sativum) were used to determine uptake of F and phytotoxicity of NaF. Concentrations in hydroponic solutions were 0–1000 mg F/L and 0–400 mg F/L in the preliminary and definitive test. A third test was done with soils collected from a fluoride-contaminated site at Fredericia, Denmark. The EC₁₀, EC₂₀ and EC₅₀-values for inhibition of transpiration were determined to 38.0, 59.6 and 128.7 mg F/L, respectively. The toxicity test with soil showed strong inhibition for the sample with the highest fluoride concentration (405 mg free F per kg soil, 75 mg F per L soil solution). The seed germination and root elongation test with cress gave EC₁₀, EC₂₀ and EC₅₀-values of 61.4, 105.0 and 262.8 mg F/L, respectively. At low external concentrations, fluoride was taken up more slowly than water and at high external concentrations at the same velocity. This indicates that an efflux pump becomes overloaded at concentrations above 210 mg F/L. Uptake kinetics were simulated with a non-linear mathematical model, and the Michaelis-Menten parameters were determined to half-saturation constant K_M near 2 g F/L and maximum enzymatic removal rate v_max at 9 g/(kg d).     

Plant Growth–Promoting Bacteria Facilitate the Growth of the Common Reed Phragmites australisin the Presence of Copper or Polycyclic Aromatic Hydrocarbons

To test whether plant growth–promoting bacteria might be useful in facilitating the growth of Phragmites australis, the common reed, in the presence of metals and organic compounds, P. australis seeds were treated with plant growth–promoting bacteria. The bacterium Pseudomonas asplenii AC was genetically transformed to express a bacterial gene encoding the enzyme 1-aminocyclopropane-1-carboxylate deaminase, and both the native and transformed bacteria were tested in conjunction with P. australis. Inoculation of seeds, which were subsequently grown in the presence of copper or creosote, with transformed P. asplenii AC significantly increased seed germination. Moreover, the addition of either native or transformed P. asplenii AC to P. australis seeds enabled the plants (shoots and roots) to attain a greater size than noninoculated plants after growth in soil in the presence of either copper or creosote.     

Comparative toxicity of ten organic chemicals to four earthworm species

Ten organic chemicals were tested for toxicity to four earthworm species: Allolobophora tuberculata, Eisenia fetida, Eudrilus eugeniae and Perionyx excavatus, using the European Economic Community's (EEC) earthworm artificial soil and contact testing procedure. The phenols were the most toxic chemicals tested, followed by the amine, substituted benzenes, halogenated aliphatic hydrocarbon, polycyclic aromatic hydrocarbon and phthalate as the least toxic chemical tested. Correlations among species within each type of test for a given chemical were extremely high, suggesting that the selection of earthworm test species does not markedly affect the assessment of a chemical's toxicity. The correlation between the two tests was low for all test species. The contact test LC50 for a given chemical cannot be directly correlated to an artificial soil test LC50 for the same earthworm species.     

A study of the impacts of Zn and Cu on two rhizobial species in soils of a long-term field experiment

Two agriculturally important species of rhizobia, Rhizobium leguminosarum biovar viciae (pea rhizobia) and R. leguminosarum bv. trifolii (white clover rhizobia), were enumerated in soils of a long-term field experiment to which sewage sludges contaminated predominantly with Zn or Cu, or Zn plus Cu, were added in the past. In addition to total soil Zn and Cu concentrations, soil pore water soluble Zn and free Zn²⁺, and soluble Cu concentrations are reported. Pea and white clover rhizobia were greatly reduced in soils containing ≥200 mg Zn kg^-1, and soil pore water soluble Zn and free Zn²⁺ concentrations ≥7 and ≥3 mg l^-1, respectively, in soils of pH 5.9–6. Copper also reduced rhizobial numbers, but only at high total soil concentrations (>250 mg kg^-1) and not to the same extent as Zn. Yields of field grown peas decreased significantly as total soil Zn, soil pore water soluble Zn and free Zn⁺² increased (R² = 0.79, 0.75 and 0.75, respectively; P < 0.001). A 50% reduction in seed yield occurred at a total soil Zn concentration of about 290 mg kg^-1, in soils of pH 5.9–6. The corresponding soil pore water soluble Zn and free Zn²⁺ concentrations were about 9 and 4 mg l^-1, respectively. Pea seed yields were not significantly correlated with total soil Cu (R² = 0.33) or soil pore water soluble Cu (R² = 0.39). Yield reductions were due to a combination of greatly reduced numbers of free-living rhizobia in the soil due to Zn toxicity, thus indirectly affecting N₂-fixation, and Zn phytotoxicity. These effects were exacerbated in slightly acidic soils due to increased solubility of Zn, and to some extent Cu, and an increase in the free Zn²⁺ fraction in soil pore water. The current United Kingdom, German and United States limits for Zn and Cu in soils are discussed in view of the current study. None of these limits are based on toxicity thresholds in soil pore water, which may have wider validity for different soil types and at different pH values than total soil concentrations. This revised version was published online in June 2006 with corrections to the Cover Date.     

Enhanced Biodegradation of Petroleum Hydrocarbons in Contaminated Soil

Soil samples taken from a contaminated site in Northern Quebec, Canada, exhibited a low capacity for biodegradation of total petroleum hydrocarbons (TPH), despite a high capacity for the mineralization of aromatic hydrocarbons and a low toxicity of soil leachates as measured by Microtox assay. Toxicity assays directly performed on surface soil, including earthworm mortality and barley seedling emergence, indicated moderate to high levels of toxicity. Soil biostimulation did not improve the removal of petroleum hydrocarbons, while bioaugmentation of soil with a developed enrichment culture increased the efficiency of hydrocarbon removal from 20.4% to 49.2%. A considerable increase in the removal of TPH was obtained in a bioslurry process, enhancing the mass transfer of hydrocarbons from soil to the aqueous phase and increasing the efficiency of hydrocarbon removal to over 70% after 45 days of incubation. The addition of ionic or nonionic surfactants did not have a significant impact on biodegradation of hydrocarbons. The extent of hydrocarbon mineralization during the bioslurry process after 45 days of incubation ranged from 41.3% to 58.9%, indicating that 62.7% to 83.1% of the eliminated TPH were transformed into CO₂ and water.     

Plant Biotests for Soil and Water Contaminated with Oil and Oil Products

Reactions of higher plants (mustard, oat, rye, lettuce, dill and barley) and microalgae (Euglena gracilis) to the contamination of soil and water with oil and oil products was studied. The germination of seeds was analyzed. The length of sprouts, dry biomass and length of plant roots, as well as the optical density of microalgal broth culture were determined. Negative effects of soil and water contamination with oil and oil products on plant and microalgal parameters examined was shown. After biological destruction of contaminants by an association of destructor strains (Acinetobactersp., Mycobacterium flavescens andRhodoccocussp.), the toxicity of contaminated mediums decreased. The data suggest that the integral toxicity of soil and water contaminated with oil and oil products and toxicity changes during biodestruction of these pollutants can be analyzed by using plant test organisms.     

Influences of addition of different krilium in saline-sodic soil on the seed germination and growth of cabbage

China is one of the largest countries with huge amount of saline-sodic soil. How to ameliorate these lands is a hotspot in China. Through measurement of soil salinity, pH value and seed germination and growth status of cabbage seedlings, 17 kinds of krilium were added to experimented soil to evaluate their effects on amelioration of heavy saline-sodic soil. Firstly, among these 17 kinds of krilium, 11 (HPMA, gypsum, HEDP, T225, PAPEMP, MA-AA, ATMP, AA/AMPS, PAA, charcoal and BHMTPMPA) were effective agents and could effectively induce seed germination without any germination phenomena in the control saline-sodic soil. HPMA was the best one in ameliorating saline-sodic soil, which has been manifested by various indices of seed germination. Secondly, addition of gypsum together with other organic agents could strongly increase the ameliorative effectiveness if compared with relatively weak agents, while decrease seed germination if compared with the stronger agent of the 2 mixed agents, showing that the synergetic effect between gypsum and other kinds of tested krilium was not evident. Thirdly, optimum dosages for separate addition of HPMA and gypsum were 42.8 L m^?3 and 25 kg m^?3, while mixed-addition of these 2 agents at the same dosages did not cause plus effect in soil improvement. Finally, field application of HPMA could enhance the growth of cabbage seedlings at the first month, but began to wilt thereafter. Thus, instead of one-time addition, several-time addition of HPMA at the suitable dosage in field practice could strengthen the effectiveness of soil amelioration.     

蚯蚓在我国南方土壤修复中的应用

蚯蚓作为生物量最大的土壤动物, 对土壤生态系统和环境质量影响深远。本研究介绍了华南地区主要应用的皮质远盲蚓(Amynthas corticis)、毛利远盲蚓(A. morrisi)、壮伟远盲蚓(A. robustus)、参状远盲蚓(A. aspergillum)、南美岸蚓(Pontoscolex corethrurus)和赤子爱胜蚓(Eisenia fetida)的生态特征, 阐述了它们与土壤pH值、酶活性、金属富集和有效性改变、孔道和微团聚体形成之间的紧密关系: (1)蚯蚓生存的土壤酸碱性范围较广(pH为3.8-7.9), 其存活率与土壤类型、有机质含量和成分、土壤污染程度和蚯蚓种类相关; (2)肠道内、蚓粪和蚓触圈的酶活性分别表征了蚯蚓取食喜好、土壤养分循环及微生物种群特征; (3)蚯蚓能够富集不同种类的金属并改变其有效性, 这些变化具有蚓种间、金属种类间和土壤类型之间的差异; (4)蚯蚓活动及其生产的蚓粪能改变土体结构、产生孔道、影响土壤团聚体数量、大小和分布。蚯蚓的上述作用使其在解决中国南方红壤酸化、土壤金属污染、茶园土壤养分不平衡、高速公路建设临时用地土壤损毁等方面具有广阔的应用前景。目前, 由于华南远盲蚓的生理特征差异研究较少, 远盲蚓繁育技术的缺乏一定程度上限制了这些蚯蚓在中型和大型尺度下应用技术的研究和推广。有必要进一步挖掘蚯蚓在土壤修复中的潜力, 进行蚯蚓主导的相关技术研发, 深入探讨其影响机制。