黑麦草对土壤中苯并[a]芘动态变化的影响

本文通过盆栽试验初步研究了黑麦草 (LoliummultiflorumLam)对污染土壤中多环芳烃苯并 [a]芘动态变化的影响。盆栽试验设计 3种苯并 [a]芘处理浓度 ,分别为 1、10、10 0mgkg-1。将苗龄为 1周的黑麦草移植于受苯并 [a]芘污染的土壤中 ,同时设置有相同的苯并 [a]芘处理浓度但不种植物的对照试验。试验在2 0m3 的控温、控光的生长室内进行 ,土壤湿度维持在田间持水量的 6 0 %。通过 12 0d的温室盆栽试验 ,观察到土壤中苯并 [a]芘的可提取浓度随着时间逐渐减少 ,种植黑麦草加快了土壤中可提取态苯并 [a]芘浓度的下降。在 1、10、10 0mgkg-1苯并 [a]芘处理浓度下 ,黑麦草生长的土壤中苯并 [a]芘的减少率分别达 82 3%、74 0 %和 5 5 9%。结果还显示 ,随盆栽时间的延长 ,黑麦草根圈土壤中多酚氧化酶含量提高 ,这可能根圈土壤中可提取态苯并 [a]芘含量降低有关。黑麦草的地上部可以积累苯并 [a]芘 ,变幅在 0 0 6～ 3 6 0mgkg-1。初步认为 ,土壤具有缓解苯并 [a]芘污染的自然本能 ,促进黑麦草生长 ,增强土壤多酚氧化酶活性 ,可提高黑麦草对苯并 [a]芘污染土壤的修复能力。     

污染土壤中苯并[a]芘的微生物共代谢修复研究

多环芳烃(PAHs)是一类普遍存在于环境中的优先具毒有机污染物,具有致突变、致癌特征,美国EPA已将16种PAHs列入优先控制有机污染物黑名单中[1],苯并[a]芘(B[a]P)是PAHs的典型代表种类之一。PAHs因疏水性、辛醇水分配系数高,而易于从水生态系统向沉积层迁移,最终造成沉积层土壤的污染。目前普遍认为PAHs在土壤中是一类难降解有机物,具有较高的稳定性,生物可降解性随着苯环数和苯环密集程度增加而降低,尤其是四环以上的PAHs常以共代谢方式进行[2-3]。共代谢(Co-metabo-lism)是指某些有机物在环境中不能作为微生物的唯一碳源与能源,必须有另外的化合物存在提供碳源与能源时     

长江三角洲地区土壤环境质量与修复研究Ⅲ.农业土壤不同粒径组分中菲和苯并[a]芘的分配特征

持久性有机污染物在土壤环境中的行为在很大程度上取决于土壤有机质对它们的吸附作用，粒径分组通常用来区分具有不同有机质组成和周转速率的有机质库。本研究利用菲（Phe）和苯并[a]芘（Bap）作为多环芳烃（PAHs）低、高环组分的代表物质，研究了它们在污染区9个农业表层土壤（0～20cm）不同粒径组分（粘粒、细粉粒、粗粉粒、细砂粒和粗砂粒）中的分配特征。结果表明，Phe在不同粒径组分中的平均含量大小顺序为粗砂粒〉细砂粒〉粘粒〉细粉粒〉粗粉粒，Bap为粗砂粒〉细砂粒〉粗粉粒〉细粉粒〉粘粒。Phe和BaP在不同粒径组分中的含量与粒径组分中有机质的含量均呈显著性正相关（P〈0．01）。不同粒径组分中的有机质对Phe富集能力的大小顺序为粗粉粒〉细粉粒〉细砂粒〉粗砂粒〉粘粒，对Bap的富集能力为粗粉粒〉粗砂粒〉细粉粒〉细砂粒〉粘粒。     

真菌对污染水稻土中苯并[a]芘共代谢降解研究

在恒温和恒定转速培养条件下,模拟生物泥浆反应器法,选择从石油污染土壤中分离出来的青霉菌、黑曲霉、白腐真菌等3种真菌,在添加不同浓度菲和邻苯二甲酸作为共存底物情况下,研究其对水稻土中苯并[a]芘（B[a]P）的共代谢降解。结果表明,未灭菌土壤对B[a]P有降解能力。当土壤中添加菲时,提高了B[a]P在土壤中的降解率,100 mg kg-1浓度菲处理的降解率显著高于200 mg kg-1浓度菲处理,邻苯二甲酸对B[a]P降解影响不大。灭菌土壤中的B[a]P几乎没有降解。添加菲及邻苯二甲酸均促进了青霉菌对B[a]P的降解,其中菲浓度为100 mg kg-1处理效果最显著。与灭菌土壤相比,接种黑曲霉提高了B[a]P的降解率,但添加菲与邻苯二甲酸却均抑制了黑曲霉菌对B[a]P的降解。白腐真菌能有效地降解B[a]P,但高浓度菲抑制了白腐真菌对B[a]P的降解,同时邻苯二甲酸对促进白腐真菌降解B[a]P的效果不明显。     

丛枝菌根对菲芘污染土壤中几种酶活性的影响

采用温室盆栽试验方法,研究了丛枝菌根真菌（AMF）Glomus mosseae和Glomus etunicatum对菲芘复合污染土壤中3种酶活性的影响。宿主植物为三叶草（Trifolium repens L.）和黑麦草（Lolium multiflorum Lam）。土样中菲和芘起始浓度分别为203.4mg·kg^-1和107.5mg·kg^-1。结果表明,植物增加了土壤多酚氧化酶、酸性磷酸酶和过氧化氢酶活性。接种Glomus mosseae和Glomus etunicatum使三叶草根际土壤多酚氧化酶活性20-60d分别增加19.6%-72.0%和29.7%-90.6%,过氧化氢酶活性分别增加3.3%-12.2%和7.8%-34.7%,酸性磷酸酶活性总体呈增加趋势;接种Glomus mosseae使黑麦草根际多酚氧化酶活性增加18.0%-43.1%,过氧化氢酶活性总体上呈降低趋势,酸性磷酸酶活性先升高后降低。供试的两种AMF对三叶草根际酸性磷酸酶和过氧化氢酶活性的影响30d前有显著差异。AM真菌作用的性质与宿主植物特性有关。     

土壤pH、水分及温度对长期污染土壤中苯并[a]芘动态变化的影响初探

以受多环芳烃长期污染农田土壤中苯并[a]芘(B[a]P)为研究对象,采用室内模拟试验研究了pH、水分和温度对土壤中B[a]P的动态变化的影响。结果表明,供试污染土壤中B[a]P能被土著微生物快速去除,其消减过程受到pH、水分和温度的影响。初始pH为4.6、5.9和7.3的泥浆中B[a]P在7天内的去除率分别为43.8%、37.8%和14.0%。在土壤水分为22%、43%、65%和87%田间持水量,温度为28℃条件下培养80天后土壤中B[a]P的去除率分别为42.5%、96.6%、96.3%和34.3%。在土壤pH为4.5,土壤水分为60%±5%田间持水量,温度为25℃和40℃条件下土壤中B[a]P的半衰期分别为33天和16天。可见,pH、水分和温度等环境条件对土壤中B[a]P的消减过程具有明显的影响,可通过人为调控强化B[a]P污染土壤的生物修复。     

高锰酸钾氧化修复污染土壤中菲和芘的研究

利用人工污染土壤,研究了高锰酸钾对4种不同土壤中菲和芘的氧化修复效果。结果表明,当高锰酸钾浓度为33.33mmol·L^-1时,土壤中菲和芘的氧化去除率达到最大。高锰酸钾氧化去除率不仅与高锰酸钾浓度有关,还与土壤性质和老化时间有关。土壤有机质含量的增加会降低高锰酸钾对土壤中菲和芘的氧化去除率;随着老化时间的增加,高锰酸钾的氧化去除率逐渐降低。老化40d后,4种土壤中菲和芘的氧化去除率显著降低,菲的氧化去除率在14%～67%之间,芘的氧化去除率在61%～84%之间。高锰酸钾氧化前后,4种土壤中有机质含量下降范围为0.77%～9.21%。从土壤有机质含量来看,高锰酸钾氧化修复多环芳烃污染土壤对土壤质量影响较小,具有较好的应用前景。     

三叶草对污染土壤中芘的去除研究

Phytoremediation has been used as an emerging technology for remediation of soil contamination with polycyclic aromatic hydrocarbons (PAHs), ubiquitous persistent environmental pollutants derived from natural and anthropogenic processes, in the last decade. In this study, a pot experiment was conducted to investigate the potential of phytoremediation of pyrene from spiked soils planted with white clover (Trifolium repens) in the greenhouse with a series of pyrene concentrations ranging from 4.22 to 365.38 mg kg^-1. The results showed that growth of white clover on pyrenecontaminated soils was not affected. The removal of pyrene from the spiked soils planted with white clover was obviously higher than that from the unplanted soils. At the end of the experiment (60 d), the average removal ratio of pyrene in the spiked soils with white clover was 77%, which was 31% and 57% higher than those of the controls with or without micobes, respectively. Both roots and shoots of white clover took up pyrene from the spiked soils and pyrene uptake increased with the soil pyrene concentration. However, the plant-enhanced dissipation of soil pyrene may be the result of plant-promoted microbial degradation and direct uptake and accumulation of pyrene by white clover were only a small part of the pyrene dissipation. Bioconcentration factors of pyrene (BCFs, ratio of pyrene, on a dry weight basis, in the plant to that in the soil) tended to decrease with increase in the residual soil pyrene concentration. Therefore, removal of pyrene in the contaminated soils was feasible using white clove.     

苏丹草对土壤中菲芘的修复作用

采用盆栽试验法,研究了苏丹草对土壤中菲、芘的去除效果,及植物和微生物在去除土壤菲、芘中的交互效应。结果显示,在试验浓度范围（0-322．06mg·kg^-1内,土壤～苏丹草系统（TR,）对菲、芘的去除效果明显。种植苏丹草60d后,土壤菲、芘去除率分别为73．07％-83．92％、63％～77．62％;平均去除率分别比对照1（无植物,不加0．1％NaN3）高55．58％、50．71％,比对照2（无植物,加0．1％NaN3）高72．71％、66．57％,说明种植苏丹草可以促进微生物对土壤中芘、菲降解。土壤酶活性测定结果也显示,酶活性越高,污染物降解率越高,反之亦然。因此,植物一微生物间的交互效应是土壤中多环芳烃降解的主要途径。     

土壤苯并[a]芘多次叠加污染对蚯蚓体腔细胞抗氧化酶的毒性效应

苯并[a]芘(B[a]P)是具有典型"三致"效应的持久性有机污染物,在土壤中逐步累积,对土壤环境质量造成潜在的威胁。一次污染条件下,B[a]P进入土壤的过程与较低剂量逐步累积的污染过程存在一定差异,在一定程度上会高估B[a]P的环境风险。本试验采用一次污染和多次累积污染2种方法,模拟污染物在土壤中逐步累积的过程,研究土壤B[a]P叠加污染的有效性特征及其在蚯蚓(Eisenia foetida)体内的富集规律,分析蚯蚓体腔细胞超氧化物歧化酶(SOD)和过氧化物酶(POD)的毒性效应。结果表明,随着土壤中B[a]P培养时间的延长,B[a]P有效含量、蚯蚓富集量和蚯蚓体腔细胞SOD、POD活性均呈逐渐下降的趋势。在多次叠加污染和一次污染条件下,土壤B[a]P有效含量在培养前期(1~28 d)下降速率分别为2.37μg·kg-1·d-1和3.35μg·kg-1·d-1,后期(28~56 d)下降速率逐步减缓为0.24μg·kg-1·d-1和0.53μg·kg-1·d-1;蚯蚓体内B[a]P富集量在培养前期(1~28 d)下降速率分别为6.94μg·kg-1·d-1和14.84μg·kg-1·d-1,后期(28~56 d)逐步减缓为0.73μg·kg-1·d-1和1.64μg·kg-1·d-1。蚯蚓体内B[a]P富集量与土壤B[a]P有效含量(Tenax吸附提取量)之间呈极显著正相关(P0.01),相关系数(R2)为0.914 7。蚯蚓体腔细胞SOD和POD活性与土壤B[a]P有效含量和蚯蚓体内B[a]P富集量之间分别呈极显著正相关(P0.01),相关系数(R2)分别为0.754 3(SOD)、0.829 6(POD)和0.704 0(SOD)、0.727 1(POD)。在叠加污染条件下,土壤B[a]P有效含量比一次污染低17.1%~38.6%(P0.05),蚯蚓富集量比一次污染低22.6%~46.8%(P0.05),蚯蚓体腔细胞SOD和POD活性分别为一次污染酶活性的49.6%~82.7%和75.5%~109.6%,差异达到显著水平(P0.05)。这表明土壤B[a]P多次叠加污染对蚯蚓体腔细胞的毒性效应低于一次污染。     

Phenylpyrazoles impact on Folsomia candida (Collembola)

Effects of the broad-spectrum insecticide fipronil were investigated on a non-target insect living in the soil, the springtail Folsomia candida Willem. Fipronil induced a significant reduction in juvenile production (PNEC = 250 μg kg⁻¹ dry soil), which seemed to be linked with an impact on the first stages of springtail development: juveniles and 7-day-old adults. These young organisms have a thinner integument, a smaller mass body and a weaker detoxification efficiency and were more sensitive than adults (14 days old) to fipronil and phenylpyrazole derivatives. Contact toxicity for juveniles was measured (LC₅₀(96 h)) giving the following values: fipronil, 450 μg l⁻¹; sulfone-fipronil, 430 μg l⁻¹; sulfide-fipronil, 160 μg l⁻¹. F. candida organisms were able to avoid contaminated food because phenylpyrazoles decreased food appetency. However, F. candida could bioaccumulate fipronil through trans-tegumental penetration (BAF_96 h = 160) and its high biotransformation rate inside springtail bodies (1 ng fipronil metabolized day⁻¹ individual⁻¹) was suspected to increase this process. Under natural conditions, phenylpyrazoles risk assessment on springtails seems to be weak due to their capacity of avoiding high contaminated zones and their biochemical tolerance to this class of insecticides.     

Growth of the collembolan Folsomia candida Willem in soil supplemented with glucose

Freely available glucose improves the conditions for soil microorganisms which are utilized as food by Collembola. We examined the effects of glucose application on collembolan (Folsomia candida Willem) growth and on several biotic and abiotic soil parameters (microbial biomass, soil respiration, qCO₂, dissolved organic carbon, inorganic nitrogen, and Olson-P) in an artificial system without predatory pressure on Collembola. Glucose addition increased soil respiration and qCO₂, and decreased nutrient levels in the soil. Collembolan growth increased with increasing glucose doses. We conclude that the availability of carbon substrates can sustain collembolan growth via an improvement of microbial growth conditions.     

Toxicity of lead pollution to the collembolan Folsomia candida in Ferri-Udic Cambosols

Toxic effect of lead(Pb) pollution on Collembola in soils has seldom been studied in depth, which is especially true for growth responses since the juveniles are very small and numerous. A single species test was conducted using the collembolan Folsomia candida as the indicator species, and soil Pb pollution(at Pb concentrations of 0, 300, 600, 1 200, 2 400, and 4 800 mg kg~(-1)) was simulated by mixing Pb(NO_3)_2 with a natural unpolluted soil. Adult survival, reproduction, body length, and body Pb concentration were determined. Adult survival and reproduction started to decrease at soil Pb concentrations of 1 200 and 600 mg kg~(-1), respectively. Lead concentrations in adult and juvenile bodies increased with increasing Pb concentration in soil. The percentage and individual abundances based on body length analysis showed that the larger adults(2.1–2.6 mm, P 0.01) were more sensitive to soil Pb concentration than the intermediate(1.9–2.1 mm, P 0.001) and smaller(1.7–1.9 mm, P 0.001) adults. Similarly, the larger juveniles(1.0–1.4 mm, P 0.001) were more sensitive to soil Pb concentration than the intermediate(0.6–1.0 mm, P 0.001) and smaller(0.3–0.6 mm, P 0.001) juveniles. For both adults or juveniles, the abundance and percentage of larger individuals declined significantly with increasing soil Pb concentration, suggesting that larger individuals were more sensitive to soil Pb pollution. Therefore, body length features, especially the percentage and abundance of larger individuals, would be potential bio-indicators of soil Pb pollution and possibly other types of soil pollution.     

Genetic variation in a subterranean arthropod (Folsomia candida) as a method to identify low-permeability barriers in an aquifer

Folsomia candida (order Collembola), a common soil arthropod, has recently been described living in a shallow (2.5–5.7 m below surface), unconsolidated aquifer in southwestern Michigan. F. candida was haplotyped from 14 wells using inter-simple sequence repeats (ISSRs) markers, and genetic variation in F. candida was used as a bioindicator to identify pore scale to mesoscale migration barriers in the aquifer. With one exception, individual haplotypes were only found in single wells. A significant relationship was identified between interpopulation genetic differentiation and geographic distance, and F. candida exhibits genetic population structuring over a very fine geographic scale (0.65 km²). Monmonier's algorithm was used to identify two most likely subsurface barriers to migration. Hypothesized barriers divided the F. candida populations into three genetically distinct groups, with two groups separated by only 15 m. These groupings are not dependent on groundwater flow direction, and are supported by a minimum spanning network and AMOVA analysis. Because F. candida movement is limited by pore size, these barriers identify local areas of low permeability in the aquifer material. The study suggests that F. candida may serve an important role in identifying geological characteristics and patterns in aquifers that may be difficult to evaluate by other means.     

Sensitivity to cadmium of the standard test species Folsomia candida compared to two other species, Onychiurus yodai and Sinella umesaoi (Collembola)

Knowing whether test species used for single-species toxicity tests are representative of other species provides valuable information because contaminant effects are often extrapolated to the community level based on these tests. To evaluate how representative the sensitivity of the standard test collembolan species Folsomia candida is for Collembola, we devised toxicity tests using the collembolan species Onychiurus yodai and Sinella umesaoi, which exhibit life-forms different from F. candida. Sensitivity to cadmium (Cd) was compared between F. candida and the novel test species. The 50% effective concentrations for reproduction were 154.7, 72.2 and 40.9 mg Cd/kg dry soil for O. yodai, F. candida and S. umesaoi, respectively. Thus, the reproductive sensitivity of F. candida to Cd was between that of the other two species. Increasing reproductive sensitivity to Cd corresponded to increasing activity levels and a more surface-dwelling life-form. Our data may facilitate the generalisation of effect data for F. candida to other species.     

Two major evolutionary lineages revealed by molecular phylogeny in the parthenogenetic collembola species Folsomia candida

In order to measure the genetic variability and determine the evolutionary relationships among strains of the parthenogenetic “standard” springtail Folsomia candida, we used Random Amplified Polymorphic DNA (RAPD-PCR) markers and determined the nucleotide sequence of the 18S and 28S rDNA genes. Both types of molecular characters were found to be polymorphic. We obtained phylogenetic trees using Direct Optimization in the dynamic homology paradigm. The trees were polarized with Isotoma viridis as an outgroup. All the trees based on one or the other type of molecular characters or based on all characters pooled together, support the hypothesis of an early divergence of two distinct lineages among the 11 strains of F. candida under study. Our results also suggest that these lineages differ in their rate of evolution and mode of diversification. The geographical origin of the studied strains was examined but we found no clear relation between the phylogenetic relationships and probable geographical origins. The early divergence of several lineages in this species should be taken into account when comparing studies on genetically different strains of this model organism. RAPD-PCR typing is an easy and efficient tool for doing such a task.     

High levels of genetic differentiation between Wolbachia-infected and non-infected populations of Folsomia candida (Collembola, Isotomidae)

The Gram-negative -Proteobacterium Wolbachia pipientis has been described as an obligate endosymbiont in many arthropod species, where it induces a variety of reproductive alterations, including parthenogenesis. Recently, this microorganism has also been detected in the parthenogenetic collembolan Folsomia candida. Here, we confirm the occurrence of the endosymbiont also in two Italian parthenogenetic populations of F. candida using ultrastructural (electron microscopy) and molecular (PCR screening on two bacterial genes) evidence. The strain isolated in the Italian populations has almost-identical gene sequences compared to that previously isolated in other populations of F. candida. In addition, we discovered a population of Folsomia cf. candida, which showed the presence of both males and females. This population is not infected by Wolbachia. A screening of two mitochondrial genes (COI and COII) showed that the bisexual population has high levels of genetic divergence in comparison with the parthenogenetic ones, even suggesting the possibility that it belongs to a different species. Furthermore, the remarkably high levels of genetic divergence between the two parthenogenetic populations suggests a possible influence of Wolbachia on inducing such differentiation, and, in the long term, speciation.     

Defensive strategies of soil fungi to prevent grazing by Folsomia candida (Collembola)

Fungi represent a major part of the living biomass in the upper soil horizon and serve as an important food source for many soil organisms. We hypothesized that certain mycelial characteristics may serve to protect fungi from grazing. Specifically, this study focused on the influence of poisonous or other repellent metabolites and crystalline structures at the hyphal surface on the feeding preference of the soil microarthropod Folsomia candida Willem. The formation of crystalline structures was studied microscopically and the content of certain metabolites such as amanitin and muscarin was investigated using analytical methods. The feeding preference of F. candida was studied in different in-vitro food choice experiments. Additionally, the palatability of the fungal isolates was estimated by the amount of egg clusters laid by F. candida and by analysing the carbon and nitrogen content of the mycelia. F. candida was repelled by fungal species with toxic metabolites or crystals on their hyphal surface, which indicates that these traits serve as feeding protection. F. candida preferred dark-pigmented fungi. Total number of egg clusters and feeding preference were not correlated. However, insects that fed on fungi without repellent characteristics laid the most eggs. The amount of carbon and nitrogen in the mycelium had no influence on feeding behaviour. We conclude that the content of repellent metabolites and crystalline structures at the hyphal surface are defensive strategies of soil fungi and strongly influence feeding preference of F. candida. Other traits such as palatability were less important. Our results help to explain collembolan feeding behaviour and interactions between soil fungi and Collembola.     

Flux of Benzo(a)pyrene to the Ground Surface and its Distribution in the Ecosystem

Benzo(a)pyrene (BP) has been investigated in bulk atmospheric deposition, moss, needles of pine and some species of vascular plants. At two remote Lithuanian sites, for 1990-1995 the flux of benzo(a)pyrene from the atmosphere to the ground surface varied between 0.3 to 4.8 μg-2 mo-1. Consequently the territory of Lithuania (65,000 km2) yearly was exposed to 624-2574 kg of carcinogen. The distribution of BP in soil and various vascular plant tissues (Trifolium tepens, Elitrygea repens, Thymus serpyllum) indicates that benzo(a)pyrene is assimilated by flora. The concentration of BP is different in various organs of vascular plants and mostly depends on the degree of soil pollution. More than 300 samples of moss, mostly Hylocomium splendens and Pleurozium schreberi were analyzed for BP. From 3.1 to 896.0 μg kg-1 of BP were measured in the moss samples. The flux of BP to the ground surface correlates well with its concentration in moss. A map of BP flux across Lithuania was created.