Manual for Soil Analysis-Monitoring and Assessing Soil Bioremediant Phần 1 ppt

Manual for Soil Analysis – Monitoring and Assessing Soil Bioremediation With 31 Figures 123... The increasing use of soil bioremediation technologies requires new cepts and methods to as

Soil Biology

Volumes published in the series

Volume 1

A Singh, O.P Ward (Eds.)

Applied Bioremediation and Phytoremediation

2004

Volume 2

A Singh, O.P Ward (Eds.)

Biodegradation and Bioremediation

2004

Volume 3

F Buscot, A Varma (Eds.)

Microorganisms in Soils: Roles in Genesis and Functions2005

Volume 4

S Declerck, D.-G Strullu, J.A Fortin (Eds.)

In Vitro Culture of Mycorrhizas

2005

Rosa Margesin

Franz Schinner (Eds.)

Manual for Soil Analysis – Monitoring and Assessing Soil Bioremediation

With 31 Figures

123

Leopold Franzens University

Prof Dr Franz Schinner

Leopold Franzens University

ISBN-10 3-540-25346-7 Springer Berlin Heidelberg New York

ISBN-13 978-3-540-25346-4 Springer Berlin Heidelberg New York

This work is subject to copyright All rights reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilm or in any other way, and storage in data banks Duplication of this publication or parts thereof is permitted only under the provisions

of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer Violations are liable for prosecution under the German Copyright Law.

Springer is a part of Springer Science + Business Media

Cover design: design&production, Heidelberg, Germany

Typesetting and production: LE-TEX Jelonek, Schmidt & Vöckler GbR, Leipzig, Germany 31/3150-YL - 5 4 3 2 1 0 - Printed on acid-free paper

The increasing use of soil bioremediation technologies requires new cepts and methods to assess the feasibility of a remediation technologyand to monitor the success of the treatment The knowledge of the reac-tion of the soil microflora to contamination facilitates the optimization of

con-biodegradation Manual of Soil Analysis – Monitoring and Assessing Soil Bioremediation differs from other books on soil analysis in that the moni-

toring and assessing of soil bioremediation are the central themes

In this comprehensive laboratory manual, sampling, pretreatment andstorage of soil, feasibility studies for soil bioremediation, and the mostimportant methods to analyze physical, chemical, and biological soil pa-rameters are presented Chapters written by experts for those involved inresearch, teaching, and routine analyses outline molecular and immunolog-ical techniques, the use of conserved internal markers, radiorespirometry,bioreporter technology, the interpretation of fatty acid profiles, soil mi-crobial and enzymatic methods, and the assessment of ecotoxicity usingbioassays Particular emphasis has been placed on the comprehensible andcomplete description of the experimental procedures The broad spectrum

of modern soil biological methods provides an excellent complementation

of traditional soil investigation and characterization Our book, however,does not claim to present all modern methods available, it rather contains

a selection of the most suitable methods for investigating contaminated

soil More biological methods can be found in our volume Methods in Soil Biology (Schinner, ¨Ohlinger, Kandeler and Margesin 1996, Springer)

We are most grateful to the authors for their excellent contributions and

to Springer, especially to Dr Jutta Lindenborn and Dr Dieter Czeschlik, forcontinuous support and cooperation We also thank Dr Ajit Varma for thepossibility to publish this book in the Soil Biology Series

Innsbruck, Austria, Rosa Margesin

January 2005 and Franz Schinner

Andreas Paetz, Berndt-Michael Wilke

1.1 Objective of Soil Sampling 1

1.1.1 Principal Objectives 1

1.1.2 Specific Objectives 4

1.2 Selection of Sampling Technique 6

1.3 Sampling Strategy 7

1.3.1 General 7

1.3.2 Preliminary Investigation 7

1.3.3 Exploratory Investigation 9

1.3.4 Main Site Investigation 9

1.3.5 Samples and Sampling Points 10

1.4 Sampling Methods 25

1.4.1 General 25

1.4.2 Type of Sample 25

1.4.3 Undisturbed Samples 27

1.4.4 Cross-Contamination 34

1.4.5 Sampling Containers 34

1.5 Pretreatment 37

1.5.1 Chemical Analysis 37

1.5.2 Physical Analysis 40

1.5.3 Biological Analysis 40

1.6 Storage of Samples 41

1.6.1 General 41

1.6.2 Specific Considerations for Biological Parameters 42

1.6.3 Preparing the Samples After Storage 44

References 44

2 Determination of Chemical and Physical Soil Properties 47 Berndt-Michael Wilke 2.1 Soil Dry Mass and Water Content 47

2.2 Water-Holding Capacity 50

VIII Contents

2.3 Bulk Density – Total Porosity 52

2.3.1 Core Method 52

2.3.2 Excavation Method 54

2.3.3 Clod Method 57

2.4 Water Retention Characteristics – Pore Size Distribution 59

2.4.1 Determination of Soil Water Characteristics Using Sand, Kaolin, and Ceramic Suction Tables 62

2.4.2 Determination of Soil Water Characteristics by Pressure Plate Extractor 65

2.5 Soil pH 68

2.6 Soil Organic Matter – Soil Organic Carbon 71

2.6.1 Dry Combustion Method 72

2.6.2 Loss On Ignition Method (LOI) 74

2.7 Soil Nutrients: Total Nitrogen 76

2.7.1 Dry Combustion Method (“Elemental Analysis”) 77

2.7.2 Modified Kjeldahl Method 79

2.8 Soil Nutrients: Inorganic Nitrogen 82

2.8.1 Extraction 83

2.8.2 Quantification of Nitrate Nitrogen 84

2.8.3 Quantification of Ammonium Nitrogen 86

2.9 Soil Nutrients: Phosphorus 87

2.9.1 Extraction of Total Phosphorus 88

2.9.2 Extraction of Labile Phosphorus 90

2.9.3 Quantification of Phosphorus 91

References 93

3 Quantification of Soil Contamination 97 Kirsten S Jørgensen, Olli Järvinen, Pirjo Sainio, Jani Salminen, Anna-Mari Suortti 3.1 General Introduction 97

3.2 Volatile Hydrocarbons 99

3.3 Hydrocarbons in the Range C10to C40 103

3.4 Polyaromatic Hydrocarbons (PAHs) 109

3.5 Heavy Metals 115

References 118

4 Immunotechniques as a Tool for Detection of Hydrocarbons 121 Gra ˙zyna A Płaza, Krzysztof Ulfig, Albert J Tien 4.1 RaPID Assay Test System 121

4.2 EnviroGard Test System 126

References 130

Contents IX

5 Feasibility Studies for Microbial Remediation

Ajay Singh, Owen P Ward, Ramesh C Kuhad

5.1 Introduction 131

5.2 Determination of Biodegradation Potential 132

5.2.1 Sampling and Soil Preparation 132

5.2.2 Selective Microbial Enrichment 134

5.2.3 Controls 135

5.2.4 Soil Microcosms 136

5.2.5 Slurry Bioreactors 137

5.2.6 Land Treatment 139

5.2.7 Composting 140

5.2.8 Scale-Up 141

5.3 Process Monitoring and Evaluation 142

5.4 Bioaugmentation 143

5.5 Effect of Surfactants 144

5.5.1 Screening of Microbial Cultures for Biosurfactant Production 145

5.5.2 Effect of Biosurfactants 146

5.5.3 Effect of Chemical Surfactants 146

5.6 Optimization of Environmental Conditions 147

5.7 Optimization of Nutritional Factors 148

5.8 Conclusions 150

References 151

6 Feasibility Studies for Microbial Remediation of Metal-Contaminated Soil 155 Franz Schinner, Thomas Klauser References 159

7 Feasibility Studies for Phytoremediation of Metal-Contaminated Soil 161 Aleksandra Sas-Nowosielska, Rafal Kucharski, Eugeniusz Malkowski 7.1 Introduction 161

7.2 Phytoextraction 161

7.2.1 Treatability Study 162

7.2.2 Full-Scale Application 166

7.2.3 Conclusions 170

7.3 Phytostabilization Potential for Soils Highly Contaminated with Lead, Cadmium and Zinc 171

7.3.1 Evaluation of Site Contaminants 171

7.3.2 Logistic Considerations 172

X Contents

7.3.3 Additives 172

7.3.4 Plants 173

7.3.5 Full-Scale Application 173

7.3.6 Effectiveness of Technology 174

7.3.7 Monitoring 174

7.3.8 Conclusions 175

References 176

8 Quantification of Hydrocarbon Biodegradation Using Internal Markers 179 Roger C Prince, Gregory S Douglas References 187

9 Assessment of Hydrocarbon Biodegradation Potential Using Radiorespirometry 189 Jon E Lindstrom, Joan F Braddock References 198

10 Molecular Techniques for Monitoring and Assessing Soil Bioremediation 201 Lyle G Whyte, Charles W Greer 10.1 General Introduction 201

10.2 Extraction and Purification of Nucleic Acids (DNA) from Soil 202

10.3 Amplification of Catabolic Genotypes and 16S rDNA Genotypes by PCR 208

10.4 DGGE Analysis Soil Microbial Communities 218

10.5 Genomics in Environmental Microbiology 226

References 228

11 Bioreporter Technology for Monitoring Soil Bioremediation 233 Steven Ripp 11.1 General Introduction 233

11.2 An Overview of Reporter Systems for Soil Bioremediation Application 235

11.3 Single Point Measurements of Soil Contaminants 241

11.4 Continuous On-Line Vapor Phase Sensing of Soil Contaminants 244

11.5 Quantification of Soil-Borne lux-Tagged Microbial Popula-tions Using Most-Probable-Number (MPN) Analysis 247

References 249

Contents XI

12 Interpretation of Fatty Acid Profiles of Soil Microorganisms 251

David B Hedrick, Aaron Peacock, David C White

12.1 Obtaining Fatty Acid Profiles from Soil Samples 251

12.2 Transforming Fatty Acid Peak Areas to Total Microbial Biomass 252

12.3 Calculation and Interpretation of Community Structure 254

12.3.1 Standard Community Structure Method 254

12.3.2 Custom Community Structure Methods 255

12.3.3 Factor Analysis 255

12.4 Calculation and Interpretation of Metabolic Stress Biomarkers 256

12.5 Naming of Fatty Acids 257

References 258

13 Enumeration of Soil Microorganisms 261 Julia Foght, Jackie Aislabie 13.1 Sample Preparation and Dilution 261

13.2 Direct (Microscopic) Enumeration 264

13.3 Enumeration by Culture in Liquid Medium (Most Probable Number Technique) 268

13.4 Enumeration by Culture on Solid Medium (Plate Count Technique) 272

References 279

14 Quantification of Soil Microbial Biomass by Fumigation-Extraction 281 Rainer Georg Joergensen, Philip C Brookes 14.1 General Introduction 281

14.2 Fumigation and Extraction 282

14.3 Biomass C 284

14.3.1 Biomass C by Dichromate Oxidation 284

14.3.2 Biomass C by UV-Persulfate Oxidation 286

14.3.3 Biomass C by Oven Oxidation 288

14.4 Biomass N 289

14.4.1 Ninhydrin-Reactive Nitrogen 289

14.4.2 Total Nitrogen 292

References 294

15 Determination of Adenylates and Adenylate Energy Charge 297 Rainer Georg Joergensen, Markus Raubuch References 302

XII Contents

16 Determination of Aerobic N-Mineralization 303

Rainer Georg Joergensen

References 306

17 Determination of Enzyme Activities in Contaminated Soil 309 Rosa Margesin 17.1 General Introduction 309

17.2 Lipase-Esterase Activity 310

17.3 Fluorescein Diacetate Hydrolytic Activity 313

17.4 Dehydrogenase Activity 316

References 319

18 Assessment of Ecotoxicity of Contaminated Soil Using Bioassays 321 Adolf Eisentraeger, Kerstin Hund-Rinke, Joerg Roembke 18.1 General Introduction: Strategy 321

18.2 Sample Preparation 323

18.3 Water-Extractable Ecotoxicity 330

18.3.1 Vibrio fischeri Luminescence-Inhibition Assay 330

18.3.2 Desmodesmus subspicatus Growth-Inhibition Assay 331

18.4 Water-Extractable Genotoxicity 332

18.4.1 The umu Test 332

18.4.2 Salmonella/Microsome Assay (Ames Test) 333

18.5 Habitat Function: Soil/Microorganisms, Soil/Soil Fauna, Soil/Higher Plants 334

18.5.1 Respiration Curve Test 334

18.5.2 Ammonium Oxidation Test 337

18.5.3 Combined Earthworm Mortality/Reproduction Test 340

18.5.4 Collembola Reproduction Test 342

18.5.5 Plant Growth Test 344

18.5.6 Test Performance for the Derivation of Threshold Values 346

18.6 Combined Performance of Bioassays and Assessment of the Results 348

18.6.1 Water-Extractable Ecotoxic Potential 348

18.6.2 Water-Extractable Genotoxicity 349

18.6.3 Assessment of the Habitat Function 350

18.6.4 Overall Assessment – Combined Strategy 353

References 355

Biotechnology Research Institute, National Research Council of Canada,

6100 Royalmount Ave., Montreal, Quebec, Canada H4P 2R2

Hedrick, David B

Hedrick Services, Knoxville, TN 37932–2575; Center for Biomarker ysis, University of Tennessee, 10515 Research Drive, Suite 300, Knoxville,Tennessee 37932–2575, USA

Anal-Hund-Rinke, Kerstin

Fraunhofer Institute for Molecular Biology and Applied Ecology, P.O Box

1260, 57377 Schmallenberg, Germany

XIV Contributors

Järvinen, Olli

Finnish Environment Institute, P.O Box 140, 00251 Helsinki, Finland

Joergensen, Rainer Georg

Department of Soil Biology and Plant Nutrition, University of Kassel, bahnhofstr 1a, 37213 Witzenhausen, Germany

Shannon & Wilson, Inc., 2355 Hill Road, Fairbanks, Alaska 99709; Institute

of Arctic Biology, University of Alaska Fairbanks, Fairbanks, Alaska 99775,USA

Malkowski, Eugeniusz

Department of Plant Physiology, Faculty of Biology and Environmental tection, University of Silesia, Jagiello˜nska 28 St, 40–032 Katowice, PolandMargesin, Rosa

Pro-Institute of Microbiology, Leopold Franzens University, Technikerstrasse

Dept of Natural Resource Sciences, McGill University, Macdonald Campus

21, 111 Lakeshore Road, St Anne de Bellevue, Quebec, Canada H9X 3V9Wilke, Berndt-Michael

Institute of Ecology, Berlin University of Technology, Franklinstrasse 29,

10587 Berlin, Germany

1 Soil Sampling and Storage

Andreas Paetz, Berndt-Michael Wilke

Whenever a volume of soil is to be characterized, it is generally not sible to examine the whole and it is therefore necessary to take samples.The samples collected should be as fully representative as possible, and allprecautions should be taken to ensure that, as far as possible, the samples

pos-do not undergo any changes in the interval between sampling and nation The sampling of multiphase systems, such as soils containing water

exami-or other liquids, gases, biological material, radionuclides, exami-or other solidsnot naturally belonging to soil (e.g., waste materials), can present specialproblems In addition, the determination of some physical soil parametersmay require so-called undisturbed soil samples for correct execution of therelevant measurement

Before any sampling program is devised, it is important that the tives be first established since they are the major determining factors, e.g.,the position and density of sampling points, time of sampling, samplingprocedures, subsequent treatment of samples and analytical requirements.The details of a sampling program depend on whether the informationneeded is the average value, the distribution, or the variability of given soilparameters

objec-Andreas Paetz: Deutsches Institut für Normung (DIN), Normenausschuss Wasserwesen (NAW), 10772 Berlin, Germany

Berndt-Michael Wilke: Institute of Ecology, Berlin University of Technology, Franklinstrasse

29, 10587 Berlin, Germany, E-mail: bmwilke@tu-berlin.de

Soil Biology, Volume 5

Manual for Soil Analysis

R Margesin, F Schinner (Eds.)

c

Springer-Verlag Berlin Heidelberg 2005

It may often be necessary to carry out an exploratory analysis program before the final objectives can be defined It is important

sampling-and-to take insampling-and-to account all relevant data from previous programs at the same

or similar locations and other information on local conditions Previouspersonal experience can also be very valuable Time and money allocated

to the design of a proper sampling program are usually well justified cause they ensure that the required information is obtained efficiently andeconomically

be-It is emphasized that complete achievement of objectives of soil tigations depends mainly on the design and execution of an appropriatesampling program The four principal objectives of soil sampling may bedistinguished as follows and are discussed below:

inves-• Sampling for determination of general soil quality

• Sampling for characterization purposes in preparation of soil maps

• Sampling to support legal or regulatory action

• Sampling as part of a hazard or risk assessmenthack

The utilization of the soil and site is of varying importance depending onthe primary objective of an investigation For example, while consideration

of past, present, and future site use is particularly relevant to sampling forrisk assessment, it is less important for soil mapping where the focus is

on description rather than the evaluation of a soil Objectives such as soilquality assessment, land appraisal, and soil monitoring take utilization intoaccount to varying degrees

The results obtained from sampling campaigns to assess soil quality formapping may indicate a need for further investigation For example, ifcontamination is detected, a need arises for identification and assessment

of potential hazards and risks

Sampling for Determination of General Soil Quality

This is typically carried out at irregular time intervals to determine thequality of the soil for a particular purpose, e.g., agriculture As such, it willtend to concentrate on factors such as nutrient status, pH, organic mattercontent, trace element concentrations, and physical factors, which provide