Impact of landspread sewage sludge and earthworm introduction on established earthworms and soil structure

Summary Sewage sludge was applied to twelve 4-m² plots in two forest (mixed hardwood, Norway spruce plantation) site and one old field site. The earthworm Eisenia fetida was introduced to half the control and half the treated plots. Earthworm populations were sampled by formalin extraction and hand-sorting five times in the year following treatment. One year after treatment, soil samples were wet-sieved and water-stable aggregate size-class arrays were determined.The dominant earthworm in the study site, Lumbricus terrestris, increased in density and mean individual biomass in response to sludge treatment in mixed hardwood and old field plots. In the Norway spruce plots, L. terrestris increased in individual biomass but decreased in density following sludge application. The density of the introduced E. fetida rapidly declined in all control plots. One year after introduction, E. fetida was found only in the sludge-treated Norway spruce plot. The introduction of E. fetida with sludge decreased the density and biomass of L. terrestris in the hardwood plots.Sludge treatment increased the percentages of 4-mm diameter water-stable aggregates in old field and hardwood plots. The addition of E. fetida with sludge in the hardwood plots generated no increase in 4-mm water-stable aggregates. In the old field, sludge + E. fetida increased the 4-mm water-stable aggregates. Little change in water-stable aggregates in response to either treatment combination was seen in the Norway spruce site.     

Influence of earthworms and leaf litter on edaphic variables in sewage-sludge-treated soil microcosms

Summary A series of 48 greenhouse soil microcosms were established and treated with combinations of sewage sludge, Acer saccharum leaves, and the earthworms Eisenia fetida and Lumbricus terrestris. The microcosms were incubated at constant moisture for 110 days. Samples were then taken and analysed for sludge depth, organic-matter content, and waterstable soil aggregates. Weights of surface leaves and weights and numbers of surviving earthworms were determined for each microcosm. L. terrestris significantly reduced sludge depth and the surface organic-matter content of microcosm soil and significantly increased percentages of 4-mm diameter water-stable aggregates. Leaf litter also significantly reduced sludge depth and increased 4-mm water-stable aggregates. E. fetida inhibited surface feeding by L. terrestris, reduced its 110-day survival rate, and inhibited the production of 4-mm water-stable aggregates in L. terrestris treatments. Numbers of E. fetida increased in L. terrestris treatments. Sludge depth, organic-matter content and water-stable aggregates were not significantly different from controls in E. fetida treatments.     

不同生境对抗虫棉绿盲蝽及其天敌发生动态的影响

于2005～2007年采用田间调查方法对不同生境抗虫棉田内绿盲蝽及其天敌的发生动态进行了研究, 试验设10种处理: 远离果园棉田、紧靠果园棉田, 密度为54 000株·hm-2的均行距棉田、密度为45 000株·hm-2的大小行距棉田, 合理化控棉田、不化控棉田, 纯作棉田和插种葵花棉田, 转双价基因抗虫棉田和转单价基因抗虫棉田.结果表明, 不同生境抗虫棉田内绿盲蝽天敌的种类基本相同, 主要为龟纹瓢虫、七星瓢虫、中华草蛉、大草蛉、华姬蝽、小花蝽、T-纹豹蛛、三突花蛛、草间小黑蛛等.不同生境抗虫棉田内绿盲蝽及其天敌种群数量差异明显.远离果园、密度为45 000株·hm-2的大小行距、合理化控能减轻转基因抗虫棉田绿盲蝽的发生, 但不利于绿盲蝽天敌的发生;种植转双价基因抗虫棉能减轻绿盲蝽的发生且对绿盲蝽天敌影响不大, 插种葵花既能增加棉田绿盲蝽天敌的数量, 又能减轻绿盲蝽的发生.因此, 转双价基因抗虫棉田插种葵花, 并远离果园、合理化控、合理密度并大小行距种植是保益灭害控制绿盲蝽的有效方法.     

A comparison of the biology of Eisenia fetida and Eisenia andrei (Oligochaeta)

Summary Specimens of locally occurring Eisenia fetida were compared with specimens of E. fetida and E. andrei obtained from France. All specimens were cultured in the laboratory under controlled conditions and growth, maturation, cocoon production, hatching success, and the number of hatchlings were compared. The results showed some differences between the two forms. The eventual worm biomass was the same for both, although the growth reats and the rate at which maturity was attained differed. E. andrei had a higher cumulative cocoon production. Although the local worms resembled E. andrei in appearance their cocoon production rate resembled that of E. fetida from France. E. andrei produced higher numbers of hatchlings per cocoon. The results show that the biology of the two species/forms is similar. The study did not provide convincing proof to substantiate a taxonomic distinction between the two forms of Eisenia.     

Chemical changes during vermicomposting (Eisenia fetida) of sheep manure mixed with cotton industrial wastes

Summary Castings of Eisenia fetida from sheep manure alone and mixed with cotton wastes analysed for their properties and chemical composition every 2 weeks for 3 months and compared with the same manures in the absence of earthworms. The results showed that earthworms accelerated the mineralization rate and converted the manures into castings with a higher nutritional value and degrees of humification. The castings obtained from manure mixed with cotton wastes exhibited good agronomic quality, suggesting that this kind of industrial residue may be used in vermicomposting.Work supported by Obra Social Caixa de Barcelona     

Growth and survival of Eisenia fetida (Sav.) (Oligochaeta: Lumbricidae) in relation to temperature,moisture and presence of Enchytraeus albidus (Henle) (Enchytraeidae)

Summary The aim of this study was to investigate the environmental requirements of Eisenia fetida (Lumbricidae) and its relation to Enchytraeus albidus (Enchytraeidae). Specimens of Eisenia fetida were cultured at two different temperatures (15°C and 25°C) and moisture conditions (50% and 80%) in presence or absence of Enchytraeus albidus. Eisenia fetida grew best at the higher temperature and amount of moisture. Enchytraeus albidus impaired the growth of Eisenia fetida at the higher temperature. When Enchytraeus albidus was present, the mortality of Eisenia fetida increased.Interaction between Eisenia fetida and Enchytraeus albidus was shown, although the mechanisms remain unclear. These species, coexisting in the same habitat (compost), appear to have slightly different niches: Eisenia fetida favours warmer, moister conditions than Enchytraeus albidus.Dedicated to the late Prof.Dr. M.S. Ghilarov     

转Bt-Cry1Ac基因棉花对烟粉虱体内几种酶活力的影响

为研究转Bt基因棉花上烟粉虱种群较常规棉上升的原因是否受到Bt毒蛋白的影响,以及是否与体内保护酶和解毒酶活力变化有关,应用转Bt基因棉花‘新棉33B’及其常规棉受体‘33’为试验对象,采用ELISA法和酶活力测定法,分别研究了取食转Bt基因棉花后烟粉虱体内Bt毒蛋白含量以及体内过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)3种保护酶和乙酰胆碱酯酶(AChE)、羧酸酯酶(CarE)和谷胱甘肽-S-转移酶(GSTs)3种解毒酶活力的变化情况。结果表明,在取食转Bt基因棉花‘新棉33B’4 h后,烟粉虱体内能检测到Cry1Ac蛋白,并且在12 h后维持在一个相对稳定的状态。取食‘新棉33B’后烟粉虱体内的SOD和GSTs活力受到显著抑制(P0.05),并且随着取食时间的延长,SOD活力逐渐下降,其中取食8 h、12 h、24 h和36 h后较取食‘33’的对照分别下降了37.8%、32.1%、32.0%和31.9%。CAT、POD和CarE活力显著提高(P0.05),并且随着取食时间的延长,酶活力逐渐上升,与对照相比,取食12 h、24 h和36 h后,CAT活力分别为取食‘33’的对照的1.54倍、1.55倍和1.42倍;POD活力分别为取食‘33’的对照的1.59倍、1.39倍和1.53倍;CarE活力分别为取食‘33’的对照的1.32倍、1.34倍和1.39倍;取食‘新棉33B’对AChE活力没有明显影响。结果提示,转Bt-Cry1Ac基因棉花对烟粉虱保护酶活力总体起到促进作用,对解毒酶活力总体影响不大。故烟粉虱体内保护酶活力的增加可能会有益于烟粉虱种群的增长,但是否起到决定性作用还有待进一步研究确定。     

Use of Avoidance Tests for Investigating Potential of the Earthworm Eisenia fetida to Improve Composting of Grass Clippings

The earthworm Eisenia fetida is the most commonly used worm for worm-supported composting of organic residues. Within the present study, the potential of E. fetida for decomposing grass clippings, an organic waste which usually causes anoxic conditions and thus insufficient degradation in the course of common composting, was investigated. To enable a thorough investigation, the substrate-related requirements of E. fetida were studied using so-called avoidance tests. These tests provide a sensitive method for evaluating the preferences and aversions of soil animals related to substrate ingredients in a sublethal range. E. fetida favored relatively moist soil with about 70% of the WHC_max and the most preferred concentration of fresh grass clippings within soil was 15% (v/v). Pretreatments of the grass clippings like silage, precomposting or inoculation with the fungi Trichoderma viride and Geotrichum klebahnii were investigated and point to an increased tolerance of the worms against pre-composted and inoculated grass whereas ensiled grass and remoistened hay was avoided. The optimum concentration of ammonium for E. fetida was 18 μg NH₄⁺-N g^?1 DW soil although the worms could withstand much higher concentrations. Lactic and acetic acid, intermediates that are quickly released from fresh lawn clippings under oxygen lacking conditions, were indicated to be the most important factors for preventing worms from tolerating higher concentrations of grass clippings.     

Stress protein response (Hsp70) and avoidance behaviour in Eisenia fetida, Aporrectodea caliginosa and Lumbricus terrestris when exposed to imidacloprid

Purpose  

The earthworm species Eisenia fetida and Eisenia andrei are commonly used in ecotoxicological standard tests. In the present study, we compared the sensitivity of E. fetida with that of two soil-dwelling earthworm species (Aporrectodea caliginosa and Lumbricus terrestris) in order to evaluate the capacity of E. fetida to predict effects of the insecticide imidacloprid. Responses were compared using two endpoints, a biochemical (changes in heat shock protein level (hsp70)) and a behavioural (avoidance behaviour).     

赤子爱胜蚓谷胱甘肽和丙二醛含量变化指示重金属污染土壤的生态毒性

为探讨赤子爱胜蚓谷胱甘肽(GSH)和丙二醛(MDA)含量变化对重金属污染土壤生态毒性的指示作用,在试验室模拟条件下,利用人工土壤和自然土壤,研究了重金属含量与蚯蚓GSH和MDA含量变化的剂量效应关系。结果显示,在人工土壤试验中,Cu可诱导蚯蚓GSH和MDA含量上升,暴露2 d、7 d、14 d后,Cu浓度与蚯蚓GSH含量的相关系数分别为0.556、0.807和0.775,与MDA含量的相关系数分别为0.760、0.672和0.544,均存在显著的剂量效应关系(p<0.05)。自然土壤试验结果显示,赤子爱胜蚓的GSH和MDA含量与土壤Cu全量的偏相关系数分别为-0.830(p<0.001)和-0.599(p<0.05),与Cd全量的偏相关系数分别为-0.697(p<0.05)和-0.690(p<0.05)。同时发现,赤子爱胜蚓GSH含量变化对土壤重金属响应的敏感性要高于MDA。     

Soil persistence of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> (Bt) toxin from transgenic Bt cotton tissues and its effect on soil enzyme activities

A silty loam soil was incubated with the leaves and stems of two transgenic Bacillus thuringiensis (Bt) cotton varieties and nontransgenic Bt cotton to study the soil persistence of the Bt toxin from the decomposing transgenic Bt cotton tissues and its effect on soil enzyme activities. The results showed that after Bt cotton tissue amendment, Bt toxin was introduced into soil upon decomposition; about 50% of the introduced Bt toxin persisted in soil for at least 56 days. No Bt toxin was detected in the nontransgenic Bt cotton-amended soil; the amount of Bt toxin was the highest in the soil treated with the residue with the higher Bt toxin content. Activities of soil urease, acid phosphomonoesterase, invertase, and cellulase were stimulated by the addition of Bt cotton tissues, whereas activity of soil arylsulfatase was inhibited. Probably cotton tissue stimulated microbial activity in soil, and as a consequence, enzyme activities of soil were generally increased. This effect can mask any negative effect of the Bt toxin on microbial activity and thus on enzyme activities.     

转基因抗虫棉对赤子爱胜蚓生长、生殖及SOD活性的影响

转基因抗虫棉给人类带来经济效益的同时,也存在生物安全性问题。蚯蚓通过影响土壤的物理和生物性质使土壤肥力发生变化。目前国内外针对转基因抗虫棉对蚯蚓的影响研究十分有限,本研究通过室内喂养试验研究了转基因抗虫棉对赤子爱胜蚓生长、生殖及超氧化物歧化酶(SOD)活性的影响。试验设置5个处理:100 g牛粪+500 g土壤(CK)、50 g转基因抗虫棉叶+550 g土壤(50T)、100 g转基因抗虫棉叶+500 g土壤(100T)、50 g非转基因抗虫棉叶+550 g土壤(50NT)、100 g非转基因抗虫棉叶+500 g土壤(100NT)。结果表明,喂养同等剂量转基因抗虫棉叶与非转基因抗虫棉叶的蚯蚓体重变化趋势基本一致,总体来说喂养100 g棉叶(100T和100NT处理)的蚯蚓体重高于喂养50 g棉叶(50T和50NT处理)和喂养牛粪(CK)的蚯蚓。在培养试验结束时,喂养100 g转基因抗虫棉叶(100T处理)蚯蚓体重下降为前一次测定时的23.8%。喂养同等剂量的转基因抗虫棉叶和非转基因抗虫棉叶的蚯蚓茧数和新产生的小蚓数无显著差异(在0 27和0 57之间变化),但喂养100 g转基因抗虫棉叶(100T处理)的小蚓数低于其他处理。在大多数培养时间内,同等剂量转基因抗虫棉叶处理对蚯蚓体内SOD活性的影响与非转基因抗虫棉叶处理差异不显著,但是在试验结束时(201 d),喂养50 g棉叶(50T和50NT处理)的蚯蚓体内SOD活性显著高于喂养100 g棉叶(100T和100NT处理)的蚯蚓。     

Effects of earthworm activity and P‐solubilizing bacteria on P availability in soil

The effects of phosphate solubilizing bacteria (Bacillus megateriumi) (PSB) and earthworms (Pheretima guillelmi and Eisenia fetida) on phosphorus (P) turnover and transformation in soil were investigated in a laboratory experiment lasting for 21 days. The treatments included soil + P. guillelmi (SW1), soil + E. fetida (SW2), soil + B. megaterium (SB), soil + P. guillelmi + B. megaterium (SBW1), soil + E. fetida + B. megaterium (SBW2), and the control with sterilized soil (control) only. The results showed that the number of B. megaterium. was enhanced in all treatments especially those with earthworms added when compared with the control. Activities of acid phosphatase (pH = 6.5) increased in all treatments, especially in the presence of P. guillelmi with or without PSB (64 % and 38 %, respectively). Significant increases in both inorganic P and water‐soluble P were observed in treatments involving B. megaterium. and earthworms when compared with the control. Inoculation of both earthworms and PSB had significant effects on microbial growth, enzymatic activity, and thus enhanced the release of available P. The dual inoculation of earthworms and bacteria further accelerated P transformation. Different performances observed for the earthworm species were probably due to their different feeding habits and physiology.     

Chemical changes of beech litter during passage through the gut of the lumbricid earthworm Eisenia fetida (SAV.)

Casts of the lumbricid earthworm species E. fetida were investigated by means of chemical analyses and ¹³C NMR spectroscopy and compared to the beech litter used as food. The casts are characterized by slightly reduced C-to-N ratios, higher ash contents and lower polysaccharide and lignin concentrations in comparison with the litter material. These differences between casts and litter are very small, ranging from 1 to 7%, and are not supported by any statistical evidence. In addition, NMR and Py-FIMS data complete the picture of E. fetida casts as almost exclusively physically altered (i.e. comminuted) plant residues.     

转Bt基因棉对土壤无脊椎动物群落结构的影响

采用干、湿漏斗分离法,运用群落生态学方法,研究了转Bt基因棉"GK12"和"新棉33B"对棉田0～15 cm土层土壤无脊椎动物群落结构的影响.结果表明,线虫和螨类为棉田土壤无脊椎动物优势类群,其频度分别为54%以上和18%以上,与亲本常规棉"泗棉3号"相比,"GK12"提高了土壤线蚓数量,但降低了某些土壤无脊椎动物类群(如同翅目、目和双翅目)的数量;两种Bt棉田中,"新棉33B"棉田的同翅目昆虫数量显著高于"GK12",蜘蛛目数量显著低于"GK12"棉田,其他土壤无脊椎动物类群的数量则基本上无显著差异."GK12"显著降低了土壤无脊椎动物类群的丰富度、多样性和均匀度;两种Bt棉中,"新棉33B"棉田土壤无脊椎动物类群的丰富度、多样性和均匀度较高.Renyi多样性指数曲线也表明,"GK12"棉田各土层土壤无脊椎动物群落的多样性均显著低于"泗棉3号"棉田;"新棉33B"棉田各土层土壤无脊椎动物群落的多样性高于"GK12",但10～15 cm土层常见类群和优势类群的多样性较低.在各土层中,土壤无脊椎动物数量、多样性指数和均匀度指数动态在不同类型棉田趋势基本相同.7～11月期间,10月是土壤无脊椎动物数量的高峰期,此时在"泗棉3号"棉田土壤无脊椎动物在0～5 cm的表层和10～15 cm的深层土壤均较多,在"GK12"和"新棉33B"棉田则集中于10～15 cm的深层土壤.11月, "泗棉3号"土壤无脊椎动物类群的多样性和均匀度呈下降趋势,"GK12"则呈上升趋势.可见,种植Bt棉"GK12"可改变某些土壤无脊椎动物类群的数量,降低类群的丰富度、多样性和均匀度;两种Bt棉田的土壤无脊椎动物群落结构也有一定的差异,"新棉33B"棉田土壤无脊椎动物群落的多样性高于"GK12".     

Reproduction of the African earthworm,Eudrilus eugeniae (Oligochaeta) — cocoons

Summary Although various authors have contributed to our knowledge of the biology of Eudrilus eugeniae some basic facts about the life cycle and reproduction are still not fully documented. Quantitative observations were made of the cocoons at 25 °C in different substrates. The incubation period for 166 cocoons was 16.89 days, based on cocoons produced by worms between the ages of 70 and 100 days. These cocoons produced a mean 2.12 hatchlings per cocoon after incubation in cattle manure, moist filter paper and distilled water. A smaller batch of cocoons incubated in cattle manure produced a mean of 2.7 hatchlings per cocoon. The hatching success of the cocoons was 84% in cattle manure, 50% in distilled water, and 48% on moist filter paper. The reproductive capabilities of E. eugeniae and Eisenia fetida were compared.     

转Bt基因棉花根际细菌与古菌群落结构分析

在一年内棉花的四个生长时期（苗期,蕾期,花铃期,吐絮期）分别采集转Bt基因抗虫棉GK12和非转基因亲本棉花泗棉3号根际土壤,以及未种植棉花的背景土壤,利用末端标记限制性片段长度多态性（T-RFLP）分析技术,分析三种土壤中细菌和古菌的16S rRNA基因片段多态性,结合克隆文库建立和测序,研究了土壤中细菌和古菌群落结构的变化.结果表明：在棉花生长的各个时期,背景土壤中细菌群落结构发生了明显的变化,生物多样性指数明显降低,古菌群落结构也有一定的变化,说明季节性变化对土壤中微生物群落产生了明显的影响.与背景土壤相比,棉花种植后根际土壤中细菌和古菌群落发生显著的变化.转基因棉花与非转基因棉花相比,根际土壤细菌和古菌的种类和种群大小的分布也发生了明显的改变.克隆文库和测序结果表明土壤中主体微生物为目前未培养的、功能特性未知的细菌和古菌,转基因棉花种植对这些细菌和古菌影响的原因、环境危害和生态风险目前尚不清楚.与古菌群落相比,棉花种植对细菌群落结构的影响较小.     

Laboratory and field evaluation of mineral oil spray for the control of citrus red mite,Panonychus citri (McGregor)

Abstract

The aim of this study was to investigate the protection role of potato proteinase inhibitor-II gene against insect and fungal pathogens of cotton. The positive integration of pinII was confirmed by polymerase chain reaction and Southern analysis in transgenic tobacco plants. In insect bioassay, the transgenic lines were more resistant to cotton bollworm (Helicoverpa armigera) and caused insect mortality and severe growth inhibition. Expression against Rhizoctonia implicated a pivotal role of this gene in defence towards pathogens. Potato proteinase inhibitor-II had shown significant expression against insect as well as fungal pathogens and suggests the role of pinII in engineering insect/pathogen-resistant plants.     

Identification of AFLP Markers that Discriminate Between Cultivated Cotton and the Hawaiian Island Endemic, <Emphasis Type="Italic">Gossypium tomentosum</Emphasis>Nuttall ex Seeman

Interspecific gene flow from crop species into wild relatives may compromise the genetic integrity of native species and in the case of transgenic crops lead to the escape of transgenes into natural populations. Approximately 72% of the upland cotton (Gossypium hirsutum L.) acreage grown in the United States in 2000 utilized transgenic cultivars. The closest relative of G. hirsutum is G. tomentosum Nuttall ex Seeman, endemic to the Hawaiian archipelago. Because these two species are fully interfertile, cultivation of transgenic G. hirsutum in the Hawaiian Islands is restricted. We investigated the possibility of identifying AFLP genetic markers that are diagnostic for each species and thus could be used in future studies to detect introgression between them. In addition, we were interested in comparing levels and geographic patterns of AFLP diversity in G. tomentosum to previous estimates using allozyme data. AFLP analysis led to the detection of 11 and 16 species-specific markers for G. tomentosum and G. hirsutum, respectively. These species-specific AFLP markers will be useful for detecting gene flow between G. hirsutum and G. tomentosum that has occurred in the past and thus might occur in the future if the restrictions on cultivation of transgenic G. hirsutum are relaxed in the Hawaiian Islands. Little genetic diversity and limited geographic patterning were discovered using AFLP markers, consistent with data from previous allozyme studies.     

Effects of Single,Binary and Quinary Mixtures of Phenanthrene and Its N-PAHs on <Emphasis Type="Italic">Eisenia fetida</Emphasis> in Soil

It is now acknowledged that aromatic hydrocarbons present in contaminated soils occur in mixtures. The effect of single, binary and quinary mixtures of phenanthrene and selected nitrogen-containing polycyclic aromatic hydrocarbons (N-PAHs) were investigated on the survival, growth and behavioural index of earthworms (Eisenia fetida) over a 21-day incubation in soil. The results showed that the LC₅₀ values ranged from (not detected) ND–329.3 mg kg^?1 (single mixture), ND–219.8 mg kg^?1 (binary mixtures) to 148.4 mg kg^?1 (quinary mixture), while the EC₅₀ values (based on weight loss) ranged from 13.3–148.4 mg kg^?1 (single mixture), 63.8–148.4 mg kg^?1 (binary mixture) to 24.2 mg kg^?1 (quinary mixture). Greater impacts were recorded where N-PAHs are present with phenanthrene. Further, behavioural index of E. fetida was affected after 24-h exposure to N-PAH-amended soils. Among the N-PAHs however, benzo[h]quinoline recorded the greatest impact on the survival, growth and behavioural index of E. fetida in soil. Findings from this study showed that three ring-N-PAHs are more toxic than phenanthrene as expected from their physico-chemical properties. The binary and quinary mixtures of phenanthrene and N-PAHs in soil intensified toxicity, suggesting that PAHs-N-PAHs mixtures represent greater risk to soil biota.