武夷山国家自然保护区不同植被类型土壤微生物群落特征

运用BIOLOG微平板技术,对武夷山国家自然保护区不同植被类型土壤微生物群落结构和代谢功能多样性进行研究,探讨不同植被类型对土壤微生物群落的影响.结果表明: 不论是土壤理化性质、酶活性,还是反映土壤微生物代谢功能多样性的平均颜色变化率(AWCD),青冈林和米槠林代表的天然林均明显优于马尾松林和杉木林代表的人工林,荒地最差.AWCD随着培养时间的延长而逐渐增加,但不同植被类型土壤AWCD值具有较大差异.碳水化合物和羧酸类碳源是各植被类型土壤微生物的主要碳源,其次为氨基酸类、酚酸类和聚合物类,胺类碳源的利用率最小.土壤微生物Simpson指数、Shannon指数、丰富度指数和McIntosh指数也呈现天然林高于人工林的趋势.主成分分析表明,从31个因素中提取的与碳源利用相关的主成分1、主成分2分别能解释变量方差的56.3%和30.2%,不同植被类型土壤微生物碳源利用特征出现分异,在主成分分离中起主要贡献作用的是胺类和氨基酸类碳源.研究结果可为进一步探讨植被多样性与土壤微生物多样性之间的关系奠定基础.     

不同施肥处理下蔬菜塑料大棚土壤微生物活性及功能多样性

利用土壤酶学方法、微生物培养方法及Biolog微生物自动分析系统,分析了不同施肥处理下塑料大棚种植黄瓜与番茄的土壤中微生物活性及功能多样性.结果表明,与传统施肥相比,配方施肥下塑料大棚黄瓜土壤脱氢酶活性提高了36.5%,番茄土壤脱氢酶活性则提高了66.5%,且达到了显著水平.配方施肥下塑料大棚黄瓜与番茄土壤可培养放线菌数量分别比传统施肥处理增加了30.0%和72.2%,且都达到了显著水平.Biolog结果显示,在土壤微生物培养过程中,配方施肥下塑料大棚番茄土壤微生物群落AWCD始终大于无肥处理及传统施肥处理.与传统施肥相比,配方施肥下塑料大棚黄瓜土壤微生物培养96 h的AWCD增加了1.9%,番茄土壤微生物培养96h的AWCD则增加了68.5%,且达到了显著水平.配方施肥下塑料大棚番茄土壤微生物Shannon指数、Simpson指数及McIntosh指数都大于传统施肥处理,并且McIntosh指数达到了显著水平.PCA分析表明,配方施肥下蔬菜塑料大棚土壤微生物群落碳源利用能力与传统施肥处理明显不同.     

外来物种刺槐对土壤微生物功能多样性的影响

以延河流域不同植被区内人工刺槐(Robinia pseudoacacia)群落和乡土植物群落的土壤微生物为研究对象,利用Biolog微平板技术对土壤微生物功能多样性进行测定,分析人工引种刺槐在不同环境梯度(3个植被区)下对土壤微生物功能多样性的影响。结果表明:刺槐对不同环境梯度下的土壤微生物的影响明显不同。从草原区到森林区,刺槐林之间土壤微生物群落的平均颜色变化率(AWCD)和土壤微生物功能多样性指数均没有显著变化;但与乡土植物群落比较,草原区、森林草原区和森林区土壤AWCD分别表现为刺槐乡土植物、刺槐乡土植物、刺槐乡土植物;在草原区和森林草原区刺槐林土壤微生物群落的群落丰富度指数(S)、Shannon-Wiener指数(H)、Simpson指数(D)、Mc Intosh指数(U)均大于乡土植物,森林区刺槐林群落丰富度指数(S)、Shannon指数(H)、Simpson指数(D)均小于乡土植物;刺槐林和乡土植物群落下土壤微生物碳源利用存在差异,主要体现在对糖类、氨基酸类的利用上。PCA分析显示主成分1贡献较大的碳源有24种,在主成分分离中起主要贡献作用的是糖类、氨基酸类和羧酸类。土壤碳氮含量能影响土壤微生物功能多样性指数,土壤含水量和温湿度能够影响碳源的利用类型。刺槐对土壤微生物功能多样性的影响存在区域差异,在评价刺槐对土壤生态过程与功能的影响时必须要考虑这种空间差异性。     

不同海拔红松林土壤微生物功能多样性

为全面了解红松林土壤微生物碳源利用特点,以长白山海拔700～1100 m红松林0～5和5～10 cm表土为研究对象,采用Biolog微平板法,分析了土壤微生物功能多样性沿海拔的垂直分布特征和变化规律.结果表明: 不同海拔红松林土壤微生物功能多样性差异显著,平均每孔颜色变化率（AWCD）随培养时间延长而增加,同一深度土层的AWCD值随海拔升高而降低;Shannon、Simpson和McIntosh多样性指数也随海拔升高呈现下降趋势,且不同海拔间3个多样性指数差异显著;物种多样性和功能多样性表现出相同的变化规律.土壤微生物对6大类碳源利用强度存在差异,各海拔土壤微生物对氨基酸类碳源利用率最高,为优势碳源;主成分分析表明,不同海拔土壤微生物在碳源利用上有明显的空间分异,土壤微生物功能多样性垂直地带性差异主要体现在对碳水类、氨基酸类和羧酸类碳源的利用上,其中碳水类尤为突出.对不同海拔土壤微生物群落功能多样性聚类分析表明,样地植被组成会对土壤微生物组成和功能活性产生重要影响.     

施用韩国假单胞菌(Pseudomonas koreensis)CLP-7对连作烟田土壤质量及微生物群落功能多样性的影响

针对目前连作导至植烟土壤酸化和根茎病害发生重的问题,田间条件下,研究了嗜酸性韩国假单胞Pseudomonas koreensis CLP-7对酸化植烟土壤pH、养分、酶活性及微生物群落功能多样性的影响。结果表明:CLP-7可以提高酸化植烟土壤pH;同时,CLP-7显著提高根际土壤速效钾、有效磷、铵态氮、硝态氮和有机质含量,其中有机质含量增加最显著;根际土壤脲酶、蔗糖酶活性呈上升趋势且均高于未施菌土壤,过氧化氢酶活性变化不明显。CLP-7处理不同时间的土壤微生物代谢多样性指数表现出明显的差异,施菌30 d时Shannon指数、Simpson指数,Richness及McIntosh指数均达到最高;所有处理的Pielou指数变化较小。微生物碳代谢主成分分析结果显示,施菌30 d时,根际土壤微生物对碳源利用与其他处理差异显著,主要为羧酸类和糖类碳源;随着施菌时间增加,微生物对氨基酸类、羧酸类、双亲化合物类、聚合物类和糖类利用率明显提高,说明CLP-7有利于提高连作烟田根际土壤微生物对碳源的利用能力。综上所述,P.koreensis CLP-7能够明显提高土壤pH、土壤酶活性,增加土壤养分含量和土壤微生物群落功能多样性,进而改善酸化植烟土壤质量,在微生物修复酸化土壤和减轻根茎病害发生的烟草绿色防控中具有较大的应用潜力。     

EG03菌剂对辣椒青枯病的防治效果及对根围土壤微生物群落的影响

EG03菌剂由多种抗青枯劳尔氏菌(Ralstonia solanacearum)的拮抗细菌复配而成,田间施用能有效防治辣椒青枯病,平均防效达85.8%.采用稀释平板法、最大或然数(MPN)法以及Biolog微孔板培养系统研究了EG03菌剂对田间辣椒根围土壤微生物群落特征的影响.结果表明:EG03菌剂对根围土壤微生物群落的影响随时间变化而不同,施用后不同程度提高了真菌和芽孢杆菌的数量,并显著增加了自生固氮细菌的数量.根围土壤微生物群落平板每孔颜色平均变化率(AWCD)的变化随培养时间呈现典型的"S"型曲线,辣椒生长后期根围土壤AWCD值高于前期.6类碳源利用分析表明,EG03菌剂的施用在短期内会降低根围土壤微生物的碳源利用率,辣椒生长后期根围土壤中以糖类物质作为碳源的微生物占主导地位.微生物多样性指数分析发现,EG03菌剂施用前期会不同程度降低根围土壤微生物各项多样性指数,但施用后期会提高各项多样性指数,尤其在Simpson指数和McIntosh均匀度两项指标上差异显著.     

冬季覆盖作物秸秆还田对双季稻田根际土壤微生物群落功能多样性的影响

微生物群落功能多样性是土壤质量变化重要的指标,不同作物类型的秸秆还田措施对土壤微生物群落功能多样性具有明显的影响。以位于双季稻主产区不同冬季覆盖作物-双季稻种植模式大田定位试验田为研究对象,以冬闲-双季稻种植模式为对照(CK),应用Biolog-GN技术开展黑麦草-双季稻(Ry)、紫云英-双季稻(Mv)、油菜-双季稻(Ra)和马铃薯-双季稻(Po)种植模式条件下不同冬季覆盖作物秸秆还田后对双季稻田根际土壤微生物功能多样性影响的研究。研究结果表明,早稻和晚稻成熟期,与CK处理相比,冬季覆盖作物秸秆还田处理增加了稻田土壤碳源平均颜色变化率(AWCD),以Po处理AWCD均为最高,均显著高于Ry和CK处理。不同冬季覆盖作物秸秆还田处理土壤微生物代谢多样性指数表现出明显的差异,早稻成熟期,Po处理的Richness、Shannon和McIntosh指数均为最高,其次为Ry、Mv和Ra处理,CK处理最低;晚稻成熟期,各处理的Richness、Shannon和McIntosh指数大小顺序均表现为PoRaMvRyCK。土壤微生物碳源利用的主成分分析结果表明,各冬季覆盖作物秸秆还田处理根际土壤微生物利用的主要碳源为氨基酸类和糖类物质,不同处理间碳源利用类型有差异。冬季覆盖作物秸秆还田措施有利于提高双季稻田根际土壤微生物对碳源的利用能力、物种丰富度和均匀度。     

不同生态条件下油松(Pinus tabulaeformis)菌根根际土壤微生物群落

利用BIOLOG代谢指纹方法分析了陕南商南和陕北安塞不同生态条件下油松菌根根际土壤微生物群落。结果表明,安塞油松和商南油松菌根根际微生物对糖类和氨基酸类碳源较易利用,商南油松菌根根际微生物总体上代谢碳源的种类和活性远大于安塞油松,而且对同类碳源的代谢商南油松的AWCD比安塞油松均高出2倍多。安塞油松菌根根际微生物以氨基酸类代谢群为优势类群,商南油松以糖类代谢群为优势类群。微生物群落多样性指数和微生物群落主成分分析（PCA）指标均表明商南油松和安塞油松菌根根际土壤微生物群落有明显不同,起分异作用的碳源主要为糖类,其次是羧酸类和氨基酸类。商南油松菌根根际土壤微生物群落AWCD极显著高于安塞油松（P<0.01）,细菌数量显著高于安塞油松（P<0.05）,Shannon指数和丰富度指数达极显著性差异（P<0.01）,商南油松和安塞油松菌根侵染率差异不显著,但菌根生物量差异达极显著水平（P<001）。相关性分析表明,菌根生物量与丰富度指数、AWCD呈极显著正相关,与Shannon指数呈显著正相关,但是与菌根侵染率相关性不显著。在商南温暖潮湿丘陵区油松菌根根际微生物活性、群落大小和多样性高于安塞油松,在安塞黄土高原干旱区微生物群落稳定性强于商南油松。     

假臭草入侵对土壤养分与微生物群落功能多样性的影响

假臭草(Praxelis clematidea)是华南地区的主要外来入侵杂草之一,已对当地的生物多样性、农业生产和生态安全造成严重威胁。采用野外样方法研究了假臭草入侵(入侵年限4~5年)对土壤养分、土壤微生物生物量、土壤酶活性以及土壤微生物群落代谢活性、碳源利用特征与功能多样性的影响。结果表明,与土著灌木丛对照区相比,假臭草入侵使土壤有机质、全氮、碱解氮、速效钾含量显著降低,全磷、速效磷含量的变化不明显,但全钾含量显著上升;土壤微生物生物量碳、氮、磷显著下降;土壤脲酶、蛋白酶、蔗糖酶和过氧化氢酶活性亦显著降低,但土壤纤维素酶活性显著增加;假臭草入侵显著降低土壤微生物群落的功能多样性,平均孔颜色变化率(AWCD)、碳水化合物和羧酸类碳源的利用率以及土壤微生物群落的Mc Intosh指数(U)、丰富度指数(S)显著下降,Pielou均匀度指数(E)则显著上升,但两处理间的Shannon指数(H)、Simpson优势度指数(Ds)的差异不明显。这说明假臭草入侵造成土壤养分的大量损耗,土壤酶活性与土壤微生物群落功能多样性降低,土壤质量下降。     

锶污染下燕麦对土壤酶活性和微生物群落功能多样性的影响

为了评价燕麦对金属锶(Sr)污染土壤的修复效果,通过土培盆栽试验,在土壤中添加Sr,共设置6个处理:25 mg/kg Sr(CK)、100 mg/kg Sr(L)、250 mg/kg Sr(H)、25 mg/kg Sr+燕麦(CK+燕麦)、100 mg/kg Sr+燕麦(L+燕麦)和250 mg/kg Sr+燕麦(H+燕麦)。从土壤酶(转化酶、磷酸酶、蛋白酶、脲酶和脱氢酶)活性和土壤微生物群落功能多样性来评价燕麦在锶污染土壤植物修复中的作用。结果表明:燕麦显著提高了脲酶和脱氢酶活性。燕麦显著提高了锶污染土壤微生物对31种碳源的平均利用率(AWCD),并提高了锶污染土壤条件下微生物群落的Shannon多样性指数(H)、Shannon均匀度指数(E)和碳源利用丰富度指数(S),降低了Shannon优势度指数(D)。主成分分析表明种植燕麦显著增加了锶污染土壤中微生物群落功能多样性。与未种植处理(L和H)相比,L+燕麦处理中羧酸类碳源和聚合物类碳源的利用率分别增加了19%和31%,H+燕麦处理中羧酸类碳源和聚合物类碳源的利用率分别增加了24%和41%。因此,在锶污染土壤中,燕麦显著提高了土壤酶活性,改善了微生物功能多样性。     

EM堆肥对黑土耕作区中小型土壤动物群落的影响

2012年5、7和9月在哈尔滨市典型黑土农田区,采用定点试验的方法设置空白对照、低浓度、中浓度和高浓度4个处理样地,分析不同浓度EM堆肥处理对中小型土壤动物群落组成、垂直结构、季节性动态和多样性的影响.结果表明: 不同浓度EM堆肥处理下共捕获中小型土壤动物7860只,平均密度49125只·m^-2,隶属于3纲10目,划分为30个类群.EM堆肥处理增加了中小型土壤动物类群数量,低浓度处理中小型土壤动物个体密度低于空白对照样地,但随着EM堆肥浓度的增加,中小型土壤动物个体密度均有所增加;中小型土壤动物群落的类群数和个体密度的垂直分布具有表聚性特征;中小型土壤动物群落存在一定的季节变化特征;不同浓度EM堆肥处理样地中小型土壤动物群落多样性指数均有所增加,土壤有机质对这种多样性的变化影响相对较大;EM堆肥处理下中小型土壤动物对土壤环境因子的响应不同,常见类群和优势类群对土壤环境的变化有较强的适应性,而稀有类群相对敏感,受到特定的环境因子的影响.说明EM堆肥处理可在一定程度上改善黑土区农田中小型土壤动物群落结构、增加其多样性.     

Influence of biochar and compost on phytoremediation of oil-contaminated soil

The use of pyrolyzed carbon, biochar, as a soil amendment is of potential interest for improving phytoremediation of soil that has been contaminated by petroleum hydrocarbons. To examine this question, the research reported here compared the effects of biochar, plants (mesquite tree seedlings), compost and combinations of these treatments on the rate of biodegradation of oil in a contaminated soil and the population size of oil-degrading bacteria. The presence of mesquite plants significantly enhanced oil degradation in all treatments except when biochar was used as the sole amendment without compost. The greatest extent of oil degradation was achieved in soil planted with mesquite and amended with compost (44% of the light hydrocarbon fraction). Most probable number assays showed that biochar generally reduced the population size of the oil-degrading community. The results of this study suggest that biochar addition to petroleum-contaminated soils does not improve the rate of bioremediation. In contrast, the use of plants and compost additions to soil are confirmed as important bioremediation technologies.     

城市垃圾堆肥对高羊茅生长及土壤性质的影响

采用盆栽实验方法,比较了在每盆施用130 g垃圾堆肥(T1)或1.5 g化肥(T2)以及130 g垃圾堆肥与1.5 g 化肥混合施用(T3)条件下高羊茅(Festuca arundinacea L)生长及土壤性质的变化.结果表明:T1和T3处理组的土壤容重显著低于对照(不施肥)和T2处理组,土壤中的阳离子代换量及有机质、碱解氮、速效磷和速效钾含量显著高于对照及T2处理组,且T3处理组的增加幅度更大;T1和T3处理组土壤中Pb、As、Cu、Cr和Cd含量均高于对照和T2处理组,且T1处理组土壤中Pb、As、Cu和Cd含量均显著高于仍处理组,但均小于土壤环境质量一级标准,不会造成土壤环境污染.T1和T3处理组高羊茅种子萌发率均明显低于T2处理组和对照,但随萌发时间的延长逐渐提高,表明垃圾堆肥对高羊茅种子萌发的抑制作用是暂时性的.施肥处理对高羊茅生长均有明显的促进作用;T1、T2和T3处理组高羊茅的株高、地上部分和根于质量、叶片N含量和叶绿素含量总体上均显著高于对照;其中T1处理组在栽培前期高羊茅的株高、地上部分干质量和叶片N含量低于T2处理组,但在后期显著高于T2处理组;而T3处理组高羊茅的各项指标总体上均最高.结果显示:使用垃圾堆肥能明显改善土壤性质、增加土壤肥力;垃圾堆肥具有缓释效应,能改善高羊茅的生长状况、提高草坪质量,与化肥混合施用效果更佳.     

Vermicompost derived from mushroom residues improves soil C/P cycling,bacterial community, and fungal abundance

The utilization of agricultural waste organic materials through composting technology has gained significant traction in agricultural production as an effective means of crop nutrient management. However, the differences in the impact of organic amendments prepared by traditional composting and vermicomposting on soil properties still deserve further research. Based on field experiments conducted in greenhouse, compared to chemical fertilizer treatments as control, we utilized traditional compost (OF) and vermicompost (VcF) derived from agricultural organic waste edible mushroom bran and cow manure (2:8). Variations in soil physiochemical properties, activities of soil enzymes related C and P cycling, abundances and diversities of bacterial 16S rRNA and fungal ITS gene at total DNA level were analyzed. Both compost treatments enhanced soil organic carbon, soil total phosphorus, and soil available P content significantly and also increased the activities of soil α-glucosidase, β-glucosidase, acid phosphomonoesterase, and alkaline phosphomonoesterase significantly. The above results suggested that soil C and P transformations were stimulated effectively by both organic amendments. OF and VcF increased the fungal ITS absolute abundances significantly while diversity indices of soil bacterial community increased significantly under both treatments. Correlation analysis indicated that bacterial community composition was strongly correlated with several soil property indexes while fungal community composition was only significantly correlated with soil total phosphorous content. In conclusion, similar to traditional compost, vermicompost significantly improved soil nutrient cycling (especially C and P aspects). In terms of soil microbes, bacteria and fungi showed different responding mechanism to vermicompost: bacteria adjust microbial structure, while fungi tend to proliferated. In consideration of the advantages of vermicompost in technology and economic cost, it could be applied in the subsequent agricultural production more frequently.     

Siderophore cooperation of the bacterium Pseudomonas fluorescens in soil

While social interactions play an important role for the evolution of bacterial siderophore production in vitro, the extent to which siderophore production is a social trait in natural populations is less clear. Here, we demonstrate that siderophores act as public goods in a natural physical environment of Pseudomonas fluorescens: soil-based compost. We show that monocultures of siderophore producers grow better than non-producers in soil, but non-producers can exploit others'' siderophores, as shown by non-producers'' ability to invade populations of producers when rare. Despite this rare advantage, non-producers were unable to outcompete producers, suggesting that producers and non-producers may stably coexist in soil. Such coexistence is predicted to arise from the spatial structure associated with soil, and this is supported by increased fitness of non-producers when grown in a shaken soil–water mix. Our results suggest that both producers and non-producers should be observed in soil, as has been observed in marine environments and in clinical populations.     

Biocontrol of Meloidogyne incognita inciting disease in tomato by using a mixed compost inoculated with Paenibacillus ehimensis RS820

Meloidogyne spp. causes root-knot disease in tomato plants. Biological control of the disease may present economically feasible, agronomically durable and environmentally safe alternative of nematicides. A chitinolytic bacterial strain, Paenibacillus ehimensis RS820, previously isolated from the soil in Korea, produced lytic enzymes in higher amounts and inhibited the growth of phytopathogenic root-knot nematodes. Moreover, the juveniles and eggs of root-knot nematodes induced secretion of lytic enzymes by RS820 including chitinases, gelatinases and collagenases. Furthermore, mixed compost containing increased amounts of chitin and inoculated with RS820 was prepared in the present study. Use of the mixed compost not only reduced the disease caused by root-knot nematodes but also improved the plant growth. The extent of inoculation of the mixed compost with RS820 significantly influenced its ability to control the root-knot disease in tomato. The mixed compost also significantly altered the activity and density of the rhizosphere bacteria. Chitinase and gelatinase producing soil bacteria, as well as their enzyme activities, were significantly influenced by the mixed compost. The mixed compost proposed in the present study may represent a viable alternative to nematicides against the root-knot nematodes in tomato.     

Co-composting of PAH-contaminated soil with poultry manure

AIMS: To investigate the effects of co-composting poultry manure with soil contaminated with different concentrations of polycyclic aromatic hydrocarbons (PAHs), on the degradation of selected PAHs in a static-pile compost system. METHODS: Mispah form (Food & Agricultural Organisation, FAO : lithosol) soil contaminated with PAHs was co-composted with poultry manure for 19 months. The soil was mixed with wood chips in a ratio of 1:1 to improve aeration and then mixed with poultry manure in a ratio of 4:1. A data logger measured temperature monthly. Residual concentrations of selected PAHs in the compost were determined monthly by gas chromatography/flame ionisation detection (GC/FID). Moisture, pH, ash content and C:N ratios were also monitored monthly. Microbial activity was measured by measuring CO(2) evolution. CONCLUSIONS: The results obtained in this experiment have shown that co-composting poultry manure with PAH-contaminated soil is capable of removing large concentrations of high molecular weight PAH from contaminated soil to levels below 1 mg kg(-1) in 19 months. SIGNIFICANCE AND IMPACT OF THE STUDY: The paper adds to the body of knowledge necessary for the development of a cost effective technology for the remediation of soil contaminated with high molecular weight PAHs by providing information on the behaviour of selected PAHs and factors such as nutrient ratio, temperature and pH during composting.     

Variations of soil structure and microbial population in a compost amended soil

Changes in soil structure and in microbial population were recorded in a long term field experiment over the growing season of maize (June–November). Determinations were made on samples from plots which had received, for two years, the following treatments: mineral fertilizers, farmyard manure and three rates of compost. Seasonal variations were observed for the stability of soil aggregates, total porosity, pore size distribution, mycorrhizal infection and aerobic cellulolytic microorganisms. The stability of the soil aggregates changed in a similar way to that found for both mycorrhizal infection and the number of aerobic cellulolytic microorganisms. Physical characteristics were not affected in any instance by the organic dressings and microbiological populations were generally influenced only by the higher doses of compost.     

Conifer growth on residual organic soils: A greenhouse study

The effects of the addition of compost, prepared from vegetable market refuse and stomach contents of slaughtered cattle, were studied in a sandy soil contained, in pots and kept in a greenhouse environment. Comparison was made between: i) a treatment involving pots containing compost mixed with 5% soil, ii) four treatments in which increasing quantities of compost homogeneously taken from the same lot (0, 10000, 20000, 30000 kg ha⁻¹) were integrated with NPK mineral fertilizer to equalize available nutrients; iii) an untreated control. At 3, 4, and 5 years from the date of treatment, after various other crops, sunflower was planted. The yield obtained, though it fell off from year to year, was approximately double in the case of 95% compost. The other four treatments also resulted in production increases compared with the untreated control. Production was found to rise progressively with increasing quantities of compost. The improvement in soil chemistry and microbiology, as shown by analyses performed 5 years after treatment with compost, suggests that the rise in crop production may be attributed to an overall improvement in all components involved in the fertility of the soil used, in our experimental conditions.     

Persistence and mobility of a Clostridium botulinum spore population introduced to soil with spiked compost

In a recent study it could be shown that compost samples can contain Clostridium botulinum. It was investigated if C. botulinum introduced with compost into botulinum-free soil can persist and be translocated within the soil. Compost was spiked with two C. botulinum type D spore concentrations (10(3) and 10(5) spores g(-1)) and the composts were spread on an experimental site. Over a period of 939 days, samples were taken from the upper (0-5 cm) and the lower (10-30 cm) soil horizons. Physical and chemical as well as microbiological variables were measured. Clostridium botulinum spores were quantified in a culture MPN-PCR assay. On day 757 the last positive sample was obtained in the plots with the lower spore concentration (10(3) g(-1)). The bacteria were never detected in the samples taken from the lower horizons of these plots. Clostridium botulinum persisted over the whole investigation period in the plots which were treated with compost spiked with 10(5) spores g(-1). The concentrations found were between 20 and 20,000 spores g(-1) soil. The bacteria were vertically translocated and could be found in the lower soil horizons (20-2000 spores g(-1) soil) starting 70 days after the compost was spread.