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Principles of ecotoxicology Third Edition. Edition Hopkin
Digital Instant Download
Author(s): Hopkin, Stephen P.; Peakall, D. B.; Sibly, R.M.; Walker, Charles
Henley
ISBN(s): 9781439894224, 1439894221
Edition: Third edition.
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Year: 2012
Language: english
Principles of
Ecotoxicology
Third Edition
3635_C000.fm Page ii Wednesday, November 9, 2005 8:06 AM
Principles of
Ecotoxicology Third Edition

C.H. Walker • S.P. Hopkin

R.M. Sibly • D.B. Peakall

Boca Raton London New York

A CRC title, part of the Taylor & Francis imprint, a member of the
Taylor & Francis Group, the academic division of T&F Informa plc.
CRC Press
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3635_C000.fm Page v Wednesday, November 9, 2005 8:06 AM

Dedication

This third edition is dedicated to the memory of Dr. David Peakall (1931–2001)
who made important contributions to the two earlier editions
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3635_C000.fm Page vii Wednesday, November 9, 2005 8:06 AM

Contents

Preface to the Third Edition ....................................................................................................... xv

Preface to the Second Edition................................................................................................... xvii
Preface to the First Edition........................................................................................................ xix
Acknowledgments ....................................................................................................................... xxi
Introduction ............................................................................................................................... xxiii

Part 1 Pollutants and Their Fate in Ecosystems

Chapter 1 Major Classes of Pollutants........................................................................................ 3
1.1 Inorganic Ions ............................................................................................................................ 3
1.1.1 Metals ................................................................................................................................. 3
1.1.2 Anions................................................................................................................................. 6
1.2 Organic Pollutants...................................................................................................................... 7
1.2.1 Hydrocarbons ..................................................................................................................... 8
1.2.2 Polychlorinated Biphenyls (PCBs) .................................................................................... 9
1.2.3 Polychlorinated Benzodioxins (PCDDs) ......................................................................... 10
1.2.4 Polychlorinated Dibenzofurans (PCDFs) ........................................................................ 11
1.2.5 Polybrominated Biphenyls (PBBs).................................................................................. 11
1.2.6 Organochlorine Insecticides............................................................................................. 11
1.2.7 Organophosphorous Insecticides (OPs)........................................................................... 13
1.2.8 Carbamate Insecticides .................................................................................................... 14
1.2.9 Pyrethroid Insecticides..................................................................................................... 14
1.2.10 Neonicotinoids ............................................................................................................... 15
1.2.11 Phenoxy Herbicides (Plant-Growth-Regulator Herbicides) .......................................... 15
1.2.12 Anticoagulant Rodenticides ........................................................................................... 16
1.2.13 Detergents....................................................................................................................... 16
1.2.14 Chlorophenols ............................................................................................................... 16
1.2.15 Ethinylestradiol (EE2).................................................................................................... 16
1.2.16 Pharmaceuticals.............................................................................................................. 18
1.3 Organometallic Compounds .................................................................................................... 18
1.4 Radioactive Isotopes ................................................................................................................ 19
1.4.1 Introduction ...................................................................................................................... 19
1.4.2 The Nature and Intensity of the Radioactive Decay Products........................................ 19
1.4.3 Half-Lives......................................................................................................................... 20
1.4.4 Biochemistry .................................................................................................................... 20
1.5 Gaseous Pollutants................................................................................................................... 21
1.6 Summary .................................................................................................................................. 21
1.7 Further Reading ....................................................................................................................... 21
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Chapter 2 Routes by Which Pollutants Enter Ecosystems

2.1 Entry Into Surface Waters........................................................................................................ 23
2.2 Contamination of Land............................................................................................................ 26
2.3 Discharge Into the Atmosphere ............................................................................................... 27
2.4 Quantification of Release of Pollutants................................................................................... 30
2.5 Summary .................................................................................................................................. 31
2.6 Further Reading ....................................................................................................................... 31

Chapter 3 Long-Range Movements and Global Transport of Pollutants

3.1 Factors Determining Movement and Distribution of Pollutants............................................. 33
3.1.1 Polarity and Water Solubility........................................................................................... 34
3.1.2 Partition Coefficients........................................................................................................ 35
3.1.3 Vapor Pressure.................................................................................................................. 35
3.1.4 Partition Between Different Compartments of the Environment.................................... 36
3.1.5 Molecular Stability and Recalcitrant Molecules ............................................................. 36
3.2 Transport in Water ................................................................................................................... 36
3.3 Transport in Air........................................................................................................................ 38
3.4 Models for Environmental Distribution of Chemicals............................................................ 40
3.5 Summary .................................................................................................................................. 42
3.6 Further Reading ....................................................................................................................... 43

Chapter 4 The Fate of Metals and Radioactive Isotopes in Contaminated Ecosystems

4.1 Introduction.............................................................................................................................. 45
4.1.1 Localization...................................................................................................................... 45
4.1.2 Persistence........................................................................................................................ 45
4.1.3 Bioconcentration and Bioaccumulation Factors.............................................................. 46
4.1.4 Bioavailability .................................................................................................................. 46
4.1.5 Cocktails of Inorganic Pollutants .................................................................................... 46
4.2 Terrestrial Ecosystems ............................................................................................................. 46
4.2.1 Introduction ...................................................................................................................... 46
4.2.2 Metals .................................................................................................................................47
4.2.3 Radioactivity .................................................................................................................... 49
4.3 Aquatic Systems....................................................................................................................... 52
4.4 Summary .................................................................................................................................. 56
4.5 Further Reading ....................................................................................................................... 56

Chapter 5 The Fate of Organic Pollutants in Individuals and in Ecosystems

5.1 Fate Within Individual Organisms........................................................................................... 58
5.1.1 General Model.................................................................................................................. 58
5.1.2 Processes of Uptake ......................................................................................................... 59
5.1.3 Processes of Distribution ................................................................................................. 62
5.1.4 Storage.............................................................................................................................. 62
5.1.5 Metabolism....................................................................................................................... 63
5.1.6 Sites of Excretion............................................................................................................. 71
5.1.7 Toxicokinetic Models....................................................................................................... 73
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5.1.8 Toxicokinetic Models for Bioconcentration and Bioaccumulation ................................ 74

5.2 Organic Pollutants in Terrestrial Ecosystems.......................................................................... 76
5.2.1 Fate in Soils ..................................................................................................................... 76
5.2.2 Transfer Along Terrestrial Food Chains .......................................................................... 79
5.3 Organic Pollutants in Aquatic Ecosystems ............................................................................. 81
5.3.1 Pollutants in Sediments.................................................................................................... 81
5.3.2 Transfer Along Aquatic Food Chains .............................................................................. 82
5.4 Summary .................................................................................................................................. 83
5.5 Further Reading ....................................................................................................................... 83

Part 2 Effects of Pollutants on Individual Organisms

Chapter 6 Toxicity Testing
6.1 General Principles.................................................................................................................... 87
6.2 Determination of the Toxicity of Mixtures ............................................................................. 91
6.3 Toxicity Testing with Terrestrial Organisms ........................................................................... 91
6.3.1 Introduction ...................................................................................................................... 91
6.3.2 Invertebrates ..................................................................................................................... 92
6.3.3 Vertebrates ........................................................................................................................ 97
6.3.4 Plants ................................................................................................................................ 98
6.4 Toxicity Testing with Aquatic Organisms ............................................................................... 99
6.4.1 Tests Concerned with Direct Absorption from Water ..................................................... 99
6.4.2 Sediment Toxicity Tests ................................................................................................. 102
6.5 Risk Assessment .................................................................................................................... 102
6.6 Toxicity Testing in the Field.................................................................................................. 104
6.7 Alternative Methods in Ecotoxicity Testing.......................................................................... 105
6.7.1 Alternative Methods for Estimating Toxicity to Vertebrates......................................... 106
6.7.2 Alternative Methods and Strategies That Work to More Ecological End Points ......... 107
6.8 Summary ................................................................................................................................ 108
6.9 Further Reading ..................................................................................................................... 108

Chapter 7 Biochemical Effects of Pollutants

7.1 Introduction............................................................................................................................ 109
7.2 Protective Biochemical Responses ........................................................................................ 111
7.3 Molecular Mechanisms of Toxicity....................................................................................... 112
7.4 Examples of Molecular Mechanisms of Toxicity ................................................................ 114
7.4.1 Genotoxic Compounds................................................................................................... 114
7.4.2 Neurotoxic Compounds ................................................................................................. 115
7.4.3 Mitochondrial Poisons ................................................................................................... 117
7.4.4 Vitamin K Antagonists ................................................................................................... 117
7.4.5 Thyroxine Antagonists ................................................................................................... 118
7.4.6 Inhibition of ATPases..................................................................................................... 119
7.4.7 Environmental Estrogens and Androgens...................................................................... 119
7.4.8 Reactions with Protein Sulfhydryl (SH) Groups........................................................... 120
3635_C000.fm Page x Wednesday, November 9, 2005 8:06 AM

7.4.9 Photosystems of Plants .................................................................................................. 120

7.4.10 Plant-Growth-Regulator Herbicides............................................................................. 120
7.5 Summary ................................................................................................................................ 120
7.6 Further Reading ..................................................................................................................... 121

Chapter 8 Physiological Effects of Pollutants

8.1 Introduction............................................................................................................................ 123
8.2 Effects of Pollutants at the Cellular Level in Animals ......................................................... 123
8.3 Effects at the Organ Level in Animals.................................................................................. 125
8.4 Effects at the Whole Organism Level in Animals ................................................................ 127
8.4.1 Neurophysiological Effects............................................................................................ 128
8.4.2 Effects on Behavior........................................................................................................ 131
8.4.3 Reproductive Effects ...................................................................................................... 133
8.5 Effects on Plants .................................................................................................................... 135
8.6 Energy Costs of Physiological Change ................................................................................. 136
8.7 Summary ................................................................................................................................ 140
8.8 Further Reading ..................................................................................................................... 140

Chapter 9 Interactive Effects of Pollutants

9.1 Introduction............................................................................................................................ 141
9.2 Additive Effects...................................................................................................................... 142
9.3 Potentiation of Toxicity ......................................................................................................... 143
9.4 Potentiation Due to Inhibition of Detoxication..................................................................... 145
9.5 Potentiation Due to Increased Activation.............................................................................. 145
9.6 The Detection of Potentiation in the Field............................................................................ 146
9.7 Summary ................................................................................................................................ 148
9.8 Further Reading ..................................................................................................................... 148

Chapter 10 Biomarkers
10.1 Classification of Biomarkers ............................................................................................... 149
10.2 Specificity of Biomarkers .................................................................................................... 150
10.3 Relationship of Biomarkers to Adverse Effects.................................................................. 152
10.4 Discussion of Specific Biomarkers...................................................................................... 154
10.4.1 Inhibition of Esterases ................................................................................................. 154
10.4.2 The Monooxygenases .................................................................................................. 154
10.4.3 Studies on Genetic Material ........................................................................................ 155
10.4.4 Porphyrins and Heme Synthesis .................................................................................. 157
10.4.5 Induction of Vitellogenin ............................................................................................. 158
10.4.6 Behavioral Biomarkers................................................................................................. 159
10.4.7 Biomarkers in Plants.................................................................................................... 160
10.5 Role of Biomarkers in Environmental Risk Assessment .................................................... 161
10.6 Summary .............................................................................................................................. 163
10.7 Further Reading ................................................................................................................... 163
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Chapter 11 In situ Biological Monitoring

11.1 Introduction.......................................................................................................................... 165
11.2 Community Effects (Type 1 Biomonitoring) ...................................................................... 165
11.2.1 Terrestrial Ecosystems ................................................................................................. 165
11.2.2 Freshwater Ecosystems ................................................................................................ 167
11.2.3 Marine Ecosystems ...................................................................................................... 168
11.3 Bioconcentration of Pollutants (Type 2 Biomonitoring) .................................................... 169
11.3.1 Terrestrial Ecosystems ................................................................................................. 169
11.3.2 Freshwater Ecosystems ................................................................................................ 170
11.3.3 Marine Ecosystems ...................................................................................................... 173
11.4 Effects of Pollutants (Type 3 Biomonitoring)..................................................................... 174
11.4.1 Terrestrial Ecosystems ................................................................................................. 174
11.4.2 Freshwater Ecosystems ................................................................................................ 175
11.4.3 Marine Ecosystems ...................................................................................................... 175
11.5 Genetically Based Resistance to Pollution (Type 4 Biomonitoring) ................................ 176
11.6 Conclusions.......................................................................................................................... 177
11.7 Summary .............................................................................................................................. 177
11.8 Further Reading .................................................................................................................. 178

Part 3 Effects of Pollutants on Populations and Communities

Chapter 12 Changes in Numbers: Population Dynamics
12.1 Population Abundance ......................................................................................................... 182
12.2 Population Growth Rate ...................................................................................................... 182
12.3 Population Growth Rate Depends on the Properties of Individual Organisms.................. 183
12.3.1 The Life History and Population Growth Rate of the Coastal Copepod
Eurytemora affinis ................................................................................................................... 184
12.4 Density Dependence ............................................................................................................ 187
12.5 Identifying Which Factors Are Density Dependent: k-Value Analysis .............................. 188
12.6 Interactions Between Species .............................................................................................. 190
12.7 Field Studies: Three Case Studies....................................................................................... 191
12.7.1 The Decline of the Partridge ....................................................................................... 192
12.7.2 Population Studies of Pesticides and Birds of Prey in the UK .................................. 198
12.7.3 The Boxworth Project (An Experimental Analysis of the Effects of
Pesticides on Farmland) .......................................................................................................... 203
12.8 Modeling the Effects of Insecticides on Skylarks for Risk Assessment Purposes ............ 206
12.9 Summary .............................................................................................................................. 207
12.10 Further Reading ................................................................................................................. 208

Chapter 13 Evolution of Resistance to Pollution

13.1 Chronic Pollution Is Environmental Change ...................................................................... 209
13.2 The Evolutionary Process in a Constant Environment ....................................................... 210
13.3 The Evolution of Resistance When There Is a Mortality–Production Trade-Off .............. 212
13.4 Evolutionary Responses to Environmental Change ............................................................ 213
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13.5 Resistance Is Often Monogenic........................................................................................... 216

13.6 Case Studies ........................................................................................................................ 217
13.6.1 Evolution of Pesticide Resistance .............................................................................. 217
13.6.2 Evolution of Metal Tolerance in Plants....................................................................... 219
13.6.3 Evolution of Industrial Melanism................................................................................ 221
13.6.4 Evolutionary Response of Dog Whelks, Nucella lapillus, to TBT Contamination........ 224
13.6.5 Evolution of Resistance to Pollution in Estuaries....................................................... 224
13.7 Summary .............................................................................................................................. 226
13.8 Further Reading ................................................................................................................... 227

Chapter 14 Changes in Communities and Ecosystems

14.1 Introduction ......................................................................................................................... 229
14.2 Changes in Soil Processes: The Functional Approach ....................................................... 230
14.3 Changes in Composition of Communities: The Structural Approach................................ 233
14.3.1 Changes in Soil Ecosystems........................................................................................ 233
14.3.2 Acidification of Lakes and Rivers ............................................................................... 236
14.3.3 Mesocosms ................................................................................................................... 238
14.4 Global Processes .................................................................................................................. 240
14.5 Summary .............................................................................................................................. 241
14.6 Further Reading ................................................................................................................... 242

Chapter 15 Extrapolating from Molecular Interactions to Consequent Effects at the

Population Level
15.1 Introduction.......................................................................................................................... 243
15.2 The Translation of Toxic Effects Across Organizational Boundaries ................................ 244
15.2.1 From Effects at the Site of Action to Localized Cellular Disturbances..................... 244
15.2.2 From Cellular Disturbances to Effects at the Level of the Whole Organism ............ 248
15.2.3 From Effects Upon the Whole Organism to Population Effects ................................ 250
15.2.4.The Complete Causal Chain ........................................................................................ 252
15.3 Biomarker Strategies............................................................................................................ 253
15.3.1 Establishing Causality Where Pollution Already Exists ............................................. 253
15.3.2 Biomarker Strategies in the Field................................................................................ 254
15.3.3 The Problem of Controls ............................................................................................. 255
15.3.4 Selection of Biomarkers for Field Studies .................................................................. 255
15.4 Biomarkers and Environmental Risk Assessment............................................................... 256
15.5 Summary .............................................................................................................................. 257
15.6 Further Reading ................................................................................................................... 257

Chapter 16 Biomarkers in Population Studies

16.1 DDE-Induced Eggshell Thinning in Raptorial and Fish-Eating Birds............................... 259
16.2 Reproductive Failure of Fish-Eating Birds on the Great Lakes of North America ........... 264
16.3 Reproductive Failure of Molluscs Caused by Tributyl Tin ................................................ 268
16.4 Forest Spraying in Eastern Canada to Control Spruce Budworm...................................... 270
16.5 Summary .............................................................................................................................. 274
16.6 Further Reading ................................................................................................................... 274
3635_C000.fm Page xiii Wednesday, November 9, 2005 8:06 AM

Appendix: Introduction to Population Projection Matrices ................................................. 275

Glossary ....................................................................................................................................... 277

Bibliography................................................................................................................................ 283

Index ............................................................................................................................................ 305

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Preface to the Third Edition

Since publication of the second edition of Prin- tems approach and to prepare the ground for
ciples of Ecotoxicology, David Peakall has died, Chapter 16, which discusses the employment of
a serious loss to the international scientific com- biomarker strategies in field studies. Chapter 15
munity. David made a very important contribu- includes examples of neurotoxicity and endo-
tion to the two earlier editions, and this third crine disruption and discusses the role of
edition is dedicated to his memory. microarray technology in a more mechanistic
In this intervening period there have been approach to ecotoxicology.
important advances in some areas of ecotoxicol- A theme running through this new material
ogy, but disappointingly slow development in is how the concepts discussed may contribute to
others, reflecting the high cost of certain types of improved methods of environmental risk assess-
research and the difficulty of obtaining support ment. To what extent will it be possible to adopt
for some areas of study that have not been given a more mechanistic approach to risk assessment
the highest priority by funding agencies. The using biomarker assays? How far is it feasible to
objective of this third edition has been to bring use results from tests where the endpoints are
the text up to date and to strengthen the treatment not toxicity data for laboratory species but
of certain topics that we believe to be very impor- changes in populations, communities, or ecosys-
tant at the present time. In doing this, we have tems? Protocols for ecotoxicity testing are cur-
continued to follow our original purpose—to rently the subject of much debate and touch not
emphasize principles rather than practice. Other only on scientific issues but on ethical, eco-
works, which describe testing protocols and mod- nomic, and political ones as well. In recent years
ern statistical techniques for analyzing data from the issue of suffering caused to laboratory ani-
ecotoxicological studies, are referred to in the text mals by toxicity testing has become important in
and will be found in the lists for further reading Western countries.
that are given at the end of each chapter. In producing this third edition the authors
Throughout the book, small changes and gratefully acknowledge the help and advice of
additions have been made. The principal ones Professor Charles Tyler for updated data con-
are as follows: An expansion of Chapter 1 cerning recent advances on endocrine disrup-
includes further pollutants that have recently tion in fish, Dr. Diane Nacci for a much valued
received attention, plus a short section on natu- contribution to the case study on Fundulus
rally occurring poisons and the history of chem- heteroclitus, Professor Paule Vasseur, Professor
ical warfare; expansions of Chapters 12 and 13 Pavel Migula, Dr. Miroslav Nakonieczny,
include new case studies and emphasize the Professor Nico van Straalen, Dr. Laurent Lag-
importance of simple measures of abundance; adic, Dr. Steve Maund, Dr. Tom Hutchinson,
additional material in Chapter 14 has been Dr. Chris Topping, Professor Paul Jepson, and
included to say more about the effects of chem- Dr. Mark Fellowes.
icals upon communities and ecosystems, giving
particular attention to recent work with meso- C.H. Walker, R.M. Sibly,
cosms; we have added a new Chapter 15 to and S.P. Hopkin
explain in more detail the molecules-to-ecosys-
3635_C000.fm Page xvi Wednesday, November 9, 2005 8:06 AM
3635_C000.fm Page xvii Wednesday, November 9, 2005 8:06 AM

Preface to the Second Edition

When writing the first edition of Principles of mining their environmental fate. The chapter
Ecotoxicology, we were conscious of the need entitled “Toxicity Testing” has been expanded
for such a book as well as of the difficulty of to give more examples, to say more about the
writing one for a new discipline which repre- problems of testing the toxicity of mixtures and
sents a synthesis of several older ones. Since to address the currently very topical issue of
publication, there has been both encouraging alternative testing methods. Chapter 8, “Physi-
support and constructive criticism pointing to ological Effects of Pollutants,” has been
certain topics that were lightly treated in the expanded to include more on neurotoxicologi-
original. The purposes of this second edition cal effects, behavioral effects, and effects on
are to fill gaps in the original that have become plants. Chapter 9 has also been expanded to
apparent with the benefit of constructive criti- deal at greater length with additive effects of
cism and of hindsight and to update some mixtures. Chapters 10 and 15 have been
aspects of this rapidly evolving discipline. updated to describe recent developments in the
It is important to emphasize that our pur- field of biomarkers. Chapter 12 has been
pose is to explain principles. Other texts empha- expanded to include recent evidence for the
size practice. Detailed descriptions of decline of certain species of birds on farmland,
ecotoxicity testing procedures or of analytical and Chapter 14 has been enlarged to deal with
procedures lie outside the scope of the present structural changes in communities in response
text and would deflect it from its primary pur- to the action of pollutants. Summaries have now
pose. Such descriptions of practice are used been added at the end of all chapters, and the
economically and are given as examples to bibliography has been updated.
illustrate principles.
In the new edition, there has been some C.H. Walker, S.P. Hopkin,
expansion of Chapter 3 to explain the impor- R.M. Sibly, and D.B. Peakall
tance of the properties of chemicals in deter-
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3635_C000.fm Page xix Wednesday, November 9, 2005 8:06 AM

Preface to the First Edition

The origins of this book lie in the MSc course ical chemist. Over a period he moved into bio-
“Ecotoxicology of Natural Populations,” which chemistry and finally into environmental
was first taught at Reading in 1991. In recent toxicology. The last move was in keeping with
years ecotoxicology has emerged as a distinct his long-standing interest and active involve-
subject of interdisciplinary character. The struc- ment in ornithology. During the last 15 years
ture of the course reflects this, and it is taught of his scientific career, he was chief of the Wild-
by people of widely differing backgrounds life Toxicology division of the Canadian Wild-
ranging from chemistry and biochemistry life Service, where he had a major involvement
through to population genetics and ecology. in studies of the Great Lakes.
Putting the different disciplines together in an Richard Sibly applied a degree in mathe-
integrated way was something of a challenge. matics first in animal behavior and then more
Experience of teaching the course per- widely in population biology. He has particular
suaded the authors of the need for a textbook interests in life history evolution and trade-offs
which would deal with the basic principles of and in how these may be affected by environ-
such a wide-ranging subject. The intention has mental pollutants.
been to approach ecotoxicology in a broad Colin Walker originally qualified as an
interdisciplinary way, cutting across traditional agricultural chemist and was responsible for
subject boundaries. However, the nature of the chemical and biochemical studies on environ-
text is bound to reflect the experience and inter- mental pollutants at Monks Wood Experimental
ests of the authors, which will now be briefly Station during the mid-1960s, when the major
reviewed. concern was about the effects of organochlorine
Steve Hopkin is a zoologist who has insecticides. He subsequently moved to the
worked on electron microscopy and x-ray anal- University of Reading, where he has developed
ysis for his Ph.D. and later investigated the teaching and research into the molecular basis
effects of metals on soil ecology at the Univer- of toxicity, with particular reference to ecotox-
sity of Bristol. Since coming to Reading his icology.
teaching and research has focused on the role
of essential and nonessential metals in the biol- C.H. Walker, S.P. Hopkin
ogy of soil invertebrates. D.B. Peakall, and R.M. Sibly
David Peakall originally graduated as a
chemist and commenced his research as a phys-
3635_C000.fm Page xx Wednesday, November 9, 2005 8:06 AM
3635_C000.fm Page xxi Wednesday, November 9, 2005 8:06 AM

Acknowledgments

Many people have contributed to this book in George Warner. Last, but not least, Gill Bogue
all sorts of ways. Although we cannot acknowl- and Val Walker who have given invaluable sec-
edge them all, we would particularly like to retarial support.
mention our MSc students, who have contrib- We have made every effort to contact
uted much in discussion and feedback, and authors/copyright holders of works reprinted in
Amanda Callaghan, Peter Dyte, Glen Fox, Principles of Ecotoxicology. This has not been
Andy Hart, Graham Holloway, Alan McCaf- possible in every case, and we would welcome
fery, Mark Macnair, Ian Newton, Demetris correspondence from those individuals/compa-
Savva, Ken Simkiss, Nick Sotherton, and nies that we have been unable to trace.
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Introduction

The term ecotoxicology was introduced by Tru- problem of progressing from the measurement
haut in 1969 and was derived from the words of concentrations of environmental chemicals
ecology and toxicology. The introduction of this to establishing their effects at the levels of the
term reflected a growing concern about the individual, the population and the community.
effects of environmental chemicals upon spe- New disciplines frequently present prob-
cies other than man. It identified an area of lems of terminology, and ecotoxicology is no
study concerned with the harmful effects of exception. Several important terms in ecotoxi-
chemicals (toxicology) within the context of cology are used inconsistently in the literature.
ecology. Up to this time, the subject of environ- Their use in the present text will now be
mental toxicology had been principally con- explained. Both pollutants and environmental
c e r n e d w i t h t h e h a r m f u l e ff e c t s o f contaminants are regarded as chemicals that
environmental chemicals upon humans, e.g., exist at levels judged to be above those that
the effects of smoke upon urban communities. would normally occur in any particular compo-
However, environmental toxicology, in its wid- nent of the environment. This immediately
est sense, encompasses the effects of chemicals raises the question of what is to be considered
upon ecosystems as well. Thus ecotoxicology normal. With most man-made organic chemi-
is a discipline within the wider field of environ- cals, such as pesticides, the situation is simple
mental toxicology (see Calow, 1994). In the — any detectable level is abnormal as the com-
present text, it is defined as the study of harmful pounds did not exist in the environment until
effects of chemicals upon ecosystems and released by man. On the other hand, chemicals
includes effects upon individuals as well as con- such as metals, sulphur dioxide, nitrogen
sequent effects at the levels of population and oxides, polycyclic aromatic hydrocarbons
above. (PAHs), and methyl mercury are naturally
Despite the definition given above, much occurring and were present in the environment
early work answering to the description of before the appearance of human beings. In the
ecotoxicology had little ecology or toxicology nature of things, there is a variation in the con-
about it. It was concerned with the detection centration of these chemicals from place to
and determination of chemicals in samples of place and from time to time. This makes it
animals and plants. Seldom could the analytical difficult to judge their normal ranges.
results be related to the effects upon individual The distinction sometimes made between
organisms, let alone effects upon populations pollutants and contaminants raises further dif-
or communities. Analytical techniques such as ficulties. The term pollutant is taken to indicate
gas chromatography, thin-layer chromatogra- that the chemical it describes is causing actual
phy, and atomic absorption facilitated the detec- environmental harm, whereas the term contam-
tion of very low concentrations of chemicals in inant implies that the chemical is not necessar-
biota; establishing the biological significance of ily harmful. The difficulties with this distinction
these residues was a more difficult matter! One are threefold. First, there is the general toxico-
of the main themes of the present text is the logical principle that toxicity is related to dose
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(Chapter 5). Thus a compound may answer to judgment on whether it should be regarded as
the description of pollutant in one situation but a pollutant or as a contaminant.
not in another — a problem mentioned earlier. Another word that has been used inconsis-
Second, there is no general agreement about tently in the literature is the term biomarker.
what constitutes environmental harm or dam- Here, biomarkers are defined as biological
age. Some scientists would regard deleterious responses to environmental chemicals at the
biochemical changes in an individual organism individual level or below, demonstrating depar-
as harmful; others would reserve the term for ture from normal status. Biomarker responses
declines in populations. Third, the effects of may be at the molecular, cellular, or whole-
measured levels of chemicals in living organ- organism level. Some workers would regard
isms — or in their environment — are seldom population responses (changes in number or
known, yet the term pollutant is frequently gene frequency) as biomarkers. However, as the
applied to them. Judgment of this issue is made latter tend to be much longer term than the
more difficult by the possibility that there is former, it may be unwise to use the same term
potentiation of toxicity when organisms are for both. In the present text, the term biomarker
exposed to mixtures of environmental chemi- will be restricted to biological responses at the
cals. To minimize these problems of terminol- level of the whole organism or below. An
ogy, the term pollutant will be applied to important thing to emphasize about biomarkers
environmental chemicals that exceed normal is that they represent measurements of effects,
background levels and can cause harm. It would which can be related to the presence of partic-
be attractive to reserve the term for particular ular levels of environmental chemicals; they
chemicals in situations where they have been provide a means of interpreting environmental
shown to cause harm, but because of the mea- levels of pollutants in biological terms.
surement problems referred to above this usage Finally, the organic pollutants to be con-
would be too restrictive. Harm will be taken to sidered here are examples of xenobiotics (for-
include biochemical or physiological changes eign compounds). They play no part in the
that adversely affect individual organisms’ normal biochemistry of living organisms. The
birth, growth, or mortality rates. Such changes concept of xenobiotics will be discussed further
would necessarily produce population declines in Chapter 5.
were it not that other processes (e.g., density An exciting feature of ecotoxicology is that
dependence) may compensate (Chapter 12). it represents a molecules-to-ecosystems
Whether a contaminant is a pollutant there- approach that relates to the genes-to-physiolo-
fore depends on its level in the environment, on gies approach originally identified by Clarke
the organism being considered, and on whether (1975) and extensively developed in North
the organism is harmed. Thus a compound may America in the 1980s (see, for example, Feder
answer to the description of pollutant for one et al., 1987). Moreover, it analyzes experimen-
organism but not for another. Because of the tal manipulations on the largest of scales
problems in demonstrating harmful effects in (although the experiments were not designed as
the field, the terms pollutant and contaminant such). Thus metal pollution, acid rain, and the
will, to a large extent, be used synonymously application of pesticides have affected whole
because it can seldom be said that contaminants ecosystems, sometimes with dramatic conse-
have no potential to cause environmental harm quences for the populations within them. In
in any situation. The term environmental chem- ecotoxicology, the ecosystem response is stud-
ical will be used to describe any chemical that ied at all levels. Initially (see Figure 1), the
occurs in the environment, without making any molecular structures of pollutants and their
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properties and environmental fate are consid- environment, and their movement, storage, and
ered (Part 1 of this book). transformation within the environment. Thus it
Ecophysiologists generally analyze the bears a certain resemblance to toxicokinetics in
impact of pollutants on an organism’s growth classic toxicology, which is concerned with the
rates, birth rates, and death rates; indeed, as uptake, distribution, metabolism, and excretion
explained above, pollutants can adversely affect of xenobiotics by living organisms (Chapter 5).
these vital rates. This makes it desirable to The difference is one of complexity. Ecotoxi-
understand how adverse effects on vital rates cology deals with movements of pollutants in
have implications for populations (Chapters 12 air, water, soils, and sediments and through
and 13). Consequently it is, in principle, possi- food chains, with chemical transformation and
ble to evaluate pollutants quantitatively in terms biotransformation.
of their population effects. This emphasis on Part 2 deals with the effects of pollutants
vital rates as crucial intervening variables, link- upon living organisms, thus resembling toxico-
ing physiological effects to population effects, dynamics in classic toxicology. The difference
is a particular feature of this book. The is again one of complexity. Whereas toxicody-
approach is continued in Chapter 13 to consider namics focuses upon interactions between
whether and how quickly resistant genes xenobiotics and their sites of action, ecotoxi-
increase in populations. The rate at which resis- cology is concerned with a wide range of effects
tant genes increase is measured by the popula- upon individual organisms at differing organi-
tion growth rate of the population of resistant zational levels (molecular, cellular, and whole
genes. The population growth rate of resistant animal). Toxicity data obtained in the labora-
genes is a measure of their Darwinian fitness. tory are used for the purposes of risk assess-
Although this is not the conventional popula- ment. Effects of pollutants are studied in the
tion-genetic measure of fitness, it is particularly laboratory, an approach that can lead to the
useful in ecotoxicology because it (alone) development of biomarker assays (Chapter 10).
shows explicitly how the fitness of resistant The use of biomarker assays in biomonitoring
genes depends on the effects those genes have is discussed in Chapter 11, which also considers
on their carriers. To summarize, the approach some effects at the population level, thereby
taken in this book allows linkage to be made looking ahead to the final part of the text.
between the different levels of organization Part 3 addresses questions that are of the
shown in Figure 1, from molecules to physiol- greatest interest to ecologists. What effects do
ogies to populations, right through to ecosys- pollutants have at the level of population, com-
tems. This is the underlying basis for the munity, and whole ecosystem? This takes the
biomarker strategy, which seeks to measure discussion into the disciplines of population
sequences of responses to pollutants from the biology and population genetics. Whereas clas-
molecular level to the level of ecosystems sic toxicology is concerned with chemical tox-
(Chapters 10, 15, and 16). The use of biomar- icity to individuals, ecotoxicologists are
kers in biomonitoring is described in Chapter particularly interested in effects at the level of
11. These four chapters are placed at the end population community and whole ecosystem.
of their respective sections of the book. They Effects at the population level may be changes
represent the practical realization of theoretical in numbers of individuals (Chapter 12), changes
aspects described in earlier chapters. in gene frequency (as in resistance) (Chapter
The text is divided into three parts. 13), or changes in ecosystem function (e.g., soil
Part 1 describes major classes of organic nitifrication) (Chapter 14). They may be suble-
and inorganic pollutants, their entry into the thal effects (e.g., on physiology or behavior)
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rather than lethal toxicity. Sometimes they may gists, physiologists, biochemists, toxicologists,
be indirect (e.g., the decline in a predator chemists, meteorologists, soil scientists, and
because of direct chemical toxicity may lead to others. It is nevertheless a discipline with its
an increase in numbers of its prey). It is often own distinct character. Apart from the important
very difficult to establish effects of pollutants applied aspects that address current public con-
on natural populations. However, the develop- cerns, it has firm roots in basic science. Chem-
ment of appropriate biomarker assays can help ical warfare is nearly as old as life itself, and
to resolve this problem. the evolution of detoxication mechanisms by
Part 3 illustrates the truly interdisciplinary animals to avoid the toxic effects of xenobiotics
character of ecotoxicology. The study of the produced by plants is paralleled by the recent
harmful effects of chemicals upon ecosystems development of resistance by pests to pesticides
draws on the knowledge and skills of ecolo- made by humans.

Ecosystems
Community
composition
Population
changes
Whole organism
responses
Physiological
changes
Biochemical
changes

Pollutant

Increasing response time

Increasing difficulty of linkage to specific chemicals

Increasing importance

Figure 1. Schematic relationship of linkages between responses at different organizational levels.

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Pollutants and Their

1 Part

Fate in Ecosystems
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Major Classes of Pollutants

Chapter
1
Many different chemicals are regarded as pol- 1990). However, in most cases, metals become
lutants, ranging from simple inorganic ions to pollutants where human activity, mainly
complex organic molecules. In the present through mining and smelting, releases them
chapter, representatives will be identified of all from the rocks in which they were deposited
the major classes of pollutant, and their prop- during volcanic activity or subsequent erosion
erties and occurrence will be briefly reviewed. and relocates them into situations where they
These pollutants will be used as examples can cause environmental damage. In April
throughout the text. Their fate in the living envi- 1998, for example, a major spill of metal-rich
ronments will be the subject of the remainder mining waste near Donana National Park, south
of Part 1. Their effects upon individuals and west Spain, caused one of the worst environ-
ecosystems will be considered in Parts 2 and 3, mental disters ever to have happened in western
respectively. Europe (Pastor et al., 2004).
The extent to which human activity con-
tributes to global cycles of metals can be
described by the anthropogenic enrichment fac-
1.1 Inorganic Ions tor (AEF) (Table 1.1). From this table it is clear
that human activity is responsible for the major-
ity of the global movement of cadmium, lead,
1.1.1 Metals
zinc, and mercury but is relatively unimportant
in the cycling of manganese. The very high AEF
A metal is defined by chemists as being an for lead is due mostly to the widespread use
element that has a characteristic lustrous and subsequent release of lead-based additives
appearance, is a good conductor of electricity, to gasoline. For most radioactive isotopes, the
and generally enters chemical reactions as pos- AEF is 100%.
itive ions or cations. Elements considered to be metals are iden-
Although metals are usually considered as tified within the periodic system for the classi-
pollutants, it is important to recognize that they fication of all elements shown in Figure 1.1.
are natural substances. With the exception of Groups of elements sharing similar chemical
radioisotopes produced in man-made nuclear properties are contained within individual verti-
reactions (bombs and reactors), all metals have cal columns. The first two columns contain ele-
been present on the earth since its formation. ments that readily lose one or two outer
There are a few examples of localized metal electrons to yield monovalent cations (column
pollution resulting from natural weathering of 1) or divalent cations (column 2). Among these
ore bodies (e.g., Hågvar and Abrahamsen, are many of the most widespread metals found

3
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4 Principles of Ecotoxicology

TABLE 1.1
Anthropogenic Enrichment Factors (AEF) for Total Global Annual Emissions of
Cadmium, Lead, Zinc, Manganese, and Mercury in the 1980s (All Values 106 kg
year–1)*
Anthropogenic Sources Natural Sources AEF (A/T) ´
Metal (A) (industry etc.) (Volcanoes etc.) Total (T) 100 (%)
Cadmium (Cd) 8 1 9 89
Lead (Pb) 300 10 310 97
Zinc (Zn) 130 50 180 72
Manganese (Mn) 40 300 340 12
Mercury (Hg) 100 50 150 66
* From various sources.

in surface waters and in soils in their stable ionic metals). Other characteristics of iron, copper,
forms, e.g., Na+, K+, Mg2+, and Ca2+. The fol- and certain other transition elements are variable
lowing 10 columns contain what are termed valency and participation in electron transfer
transition elements, and these are also regarded reactions. Electron transfer reactions involving
as more complex than the alkali and alkali earth oxygen can lead to the production of toxic
elements that constitute the first two groups. oxyradicals, a toxicity mechanism now known
Moving from left to right through the three main to be of considerable importance in both animals
series of transition elements, the nuclei become and plants: it is now recognized that some
larger and the outer electrons show less tendency oxyradicals, such as superoxide anion (·O2–) and
to escape (i.e., to form cations) than the elements the hydroxyl radical (·OH), can cause serious
listed in columns 1 and 2. Consequently, there cellular damage. In the remaining vertical
is a tendency to share electrons with other ele- groups, as one moves from left to right, there is
ments, leading to the formation of covalent a reducing tendency to form cations. There is a
bonds and complex ions (e.g., by copper, iron, progression from metals to metalloids, the latter
cobalt, or nickel). Some of the larger atoms tend showing characteristics of both metals and non-
to retain electrons and remain in the elemental metals, until the nonmetals are reached (C, N,
state (e.g., silver and gold, the so-called noble O, P, S, Cl, Br, etc.). The final vertical column

H He

Li Be B C N O F Ne

Na Mg Al Si P S Cl Ar

K Ca Sc Ti V Cr Mn Fe Co Ni Cu Zn Ga Ge As Se Br Kr

Rb Sr Y Zr Nb Mo Tc Ru Rh Pd Ag Cd In Sn Sb Te I Xe

Cs Ba La Hf Ta W Re Os Ir Pt Au Hg Ti Pb Bi Po At Rn

Fr Ra Ac Rf Ha

Ce Pr Nd Pm Sm Eu Gd Tb Dy Ho Er Tm Yb Lu

Th Pa U Np Pu Am Cm Bk Cf Es Fm Md No Lr

Figure 1.1
Periodic table of the elements. Those considered to be metals are surrounded by bold lines. Metalloids (with properties of
metals and nonmetals) are shaded. (Reproduced from Hopkin, 1989, with permission from Elsevier Applied Science
Publishers.)
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Major Classes of Pollutants 5

contains the very stable inert gases, which have TABLE 1.2
hardly any chemical reactivity. The two horizon- Separation of Some Essential and
tal boxes below the main periodic classification Nonessential Metal Ions of Importance
contain the generally rare elements of the lan- as Pollutants into Class A (Oxygen
thanide and actinide series, which are metallic Seeking), Class B (Sulfur or Nitrogen
in character. Seeking), and Borderline Elements
The tendency to form covalent bonds Based on the Classification Scheme of
shown by metalloids, and also by metals located Nieboer and Richardson (1980)*
close to them in the periodic classification, has Class A Borderline Class B
two important toxicological consequences.
First, these elements are able to bind covalently Calcium Zinc Cadmium
to organic groups, thereby forming lipophilic Magnesium Lead Copper
compounds and ions. Some of these compounds Manganese Iron Mercury
are highly toxic, e.g., tetraalkyl lead, tributyl tin Potassium Chromium Silver
oxide, methyl mercury salts, and methylated Strontium Cobalt
forms of arsenic. Because of their lipophilicity, Sodium Nickel
their distribution within animals and plants and Arsenic
their toxic action usually differs from that of Vanadium
simple ionic forms of the same elements. Orga- * This distinction is important in determining rates of
nometallic compounds are discussed later in transport across cell membranes and sites of intracel-
Section 1.3. Second, these elements can have lular storage in metal-binding proteins and metal-con-
toxic effects by binding to nonmetallic constit- taining granules (e.g., Section 8.2).
uents of cellular macromolecules, e.g., the bind-
ing of copper, mercury, lead, and arsenic to them in an insoluble form in intracellular gran-
sulphydryl groups of proteins. ules for long-term storage or excretion in the
The term heavy metal has been used exten- feces (see Chapter 8).
sively in the past to describe metals that are Essential elements all have a window of
environmental pollutants. For a metal to be con- essentiality within which dietary concentra-
sidered heavy, it must have a density relative to tions in animals, or soil concentrations in
water of greater than five. However, the term plants, have to be maintained if the organism
heavy metal has been replaced by a classifica- is to grow and reproduce normally (Figure
tion scheme that considers their chemistry 1.2). In addition to carbon, hydrogen, oxygen,
rather than their relative density (Nieboer and and nitrogen, all animals need the seven major
Richardson, 1980, Table 1.2). This approach is mineral elements calcium, phosphorus, potas-
more logical because there are some metals that sium, magnesium, sodium, chlorine; and sul-
are not heavy but can be important environmen- fur for ionic balance and as integral parts of
tal pollutants. Aluminum, for example, which amino acids, nucleic acids, and structural com-
is a metal, has a relative density of only 1.5. pounds. Thirteen other so-called trace ele-
However, it is an extremely important pollutant ments are definitely required, namely iron,
in acidified lakes, where it becomes soluble and iodine, copper, manganese, zinc, cobalt,
is toxic to fauna. The gills of fish are particu- molybdenum, selenium, chromium, nickel,
larly susceptible to aluminum poisoning. Alu- vanadium, silicon, and arsenic. Zinc, for
minum has also been implicated in Alzheimer’s example, is an essential component of at least
disease in humans and may be deposited in the 150 enzymes; copper is essential for the nor-
brain. mal function of cytochrome oxidase; and iron
Metals are nonbiodegradable. Unlike some is part of haemoglobin, the oxygen-carrying
organic pesticides, metals cannot be broken pigment in red blood cells. Boron is required
down into less harmful components. Detoxifi- exclusively by plants. A few other elements,
cation by organisms consists of hiding active such as lithium, aluminum, fluorine, and tin,
metal ions within a protein such as metallothio- may be essential at ultratrace levels. The win-
nein (binding covalently to sulfur) or depositing dow of essentiality for some elements is very
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6 Principles of Ecotoxicology

P P

Optimum No effect
Essential Non-essential
element element

y
enc

Tox
d

fici

Tox
ic
De

ic
Ce Cne
‘Window of essentiality’

Figure 1.2
Relationships between performance (P) (growth, fecundity, survival) and concentrations of an essential (Ce) or nonessential
(Cne) element of the diet of animals. Possible deficiency effects at ultratrace levels (d) of an apparently nonessential element
may be discovered as the sensitivities of analytical techniques are improved.

narrow. Selenium, for example, was consid- 1.1.2 Anions

ered for a long time to be only a dangerous
toxin until its role in the enzyme glutathione There are some inorganic pollutants that are not
peroxidase was discovered. The dose deter- particularly toxic but which cause environmen-
mines the poison. tal problems because they are used in such large
Nonessential metals such as mercury or quantities. These include anions such as nitrates
cadmium, in addition to being toxic above cer- and phosphates.
tain levels, may also affect organisms by Nitrate fertilizers are used extensively in
inducing deficiencies of essential elements agriculture. During the growth period of crops,
through competition at active sites in biologi- most of the fertilizer applied is absorbed by
cally important molecules (Table 1.3) (see plant roots. However, when growth ceases,
Chapter 7). Such antagonism also occurs nitrate released during the decomposition of
between essential elements. A concentration dead plant material passes down through the
of only 5 μg Mo (molybdenum) g–1 in the diet soil and may enrich adjacent water courses.
of cattle is sufficient to reduce copper intake The increase in available nitrogen may cause
by 75%, which often leads to symptoms of blooms in algal populations. This effect is
copper deficiency. called eutrophication and eventually leads to
oxygen starvation as microorganisms break
TABLE 1.3 down the dead algal tissues (Nosengo, 2003).
Level of Activity of Carbonic The safe limit for nitrates in drinking
Anhydrase, Expressed Relative to water in the UK has been set at 50 parts per
That of Zinc, of Different Metals million (p.p.m.). A human health problem may
Substituted in the Protein* arise if young babies ingest bottled milk made
Normal Activity
up with nitrate-contaminated water. During
Metal (Hydration of CO2) (%)
their first few months of life, human infants
have an anaerobic stomach. The nitrates are
Zinc 100 converted to nitrites in this oxygen-poor envi-
Cobalt 56 ronment. The nitrites bind to hemoglobin and
Nickel 5 reduce its capacity to carry oxygen, and the
Cadmium 4 infant may develop blue-baby syndrome or
Manganese 4 methemoglobinemia. The problem does not
Copper 1 arise with breast-fed babies (definitely a case
Mercury 0.05
of “breast is best”!). In regions of intensive
agriculture, the 100 p.p.m. level is exceeded
* From Coleman (1967).
in water extracted from rivers or in bore holes
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Major Classes of Pollutants 7

where nitrates have leached down to aquifers. bon compounds tend to be more polar and
The problem can be solved by removing the more chemically reactive when they contain
nitrate chemically at the water treatment works functional groups such as OH, HCO, and NO2.
or by diluting the contaminated water with In these examples, the oxygen atom attracts
water from a relatively nitrate-free source. The electrons away from neighboring carbon
long-term solution is of course to reduce atoms, thereby creating a charge imbalance on
nitrate usage, and this is being done in so- the molecule. Molecules of high polarity tend
called exclusion zones around sources of water to enter into chemical and biochemical reac-
for human consumption. tions more readily than do molecules of low
Similar problems of eutrophication can polarity.
also arise with phosphates used as fertilizers. The behavior of organic compounds is
However, there is an additional source: wash- dependent upon their molecular structure —
ing powders. These have been made less resis- molecular size, molecular shape, and the pres-
tant to breakdown in recent years because of ence of functional groups being important
cooperation between soap manufacturers and determinants of metabolic fate and toxicity.
water-treatment companies. In the 1950s and Thus it is important to know the formulae of
1960s, it was common to see a huge buildup pollutants in order to understand or predict what
of foam below weirs and waterfalls down- happens to them in the living environment. The
stream of the outfalls of sewage treatment principles operating here are illustrated by
works. examples given in Chapters 5 and 7. Readers
with a limited knowledge of chemistry are
referred to the text of Manahan (1994), which
1.2 Organic Pollutants contains two useful concise chapters on basic
principles.
The great majority of compounds that contain The pollutants that will be described here
carbon are described as organic, the few excep- are predominantly man-made (“anthropo-
tions being simple molecules such as CO2 and genic’) compounds that have appeared in the
CO. Carbon has the ability to enter into the natural environment only during the last cen-
formation of a bewildering diversity of complex tury. This is a very short time in evolutionary
organic compounds, many of which provide the terms, and there has been only limited oppor-
basic fabric of living organisms. The reason for tunity for the evolution of protective mecha-
this is the tendency of carbon atoms to form nisms against their toxic effects (e.g.,
stable bonds with one another, thereby creating detoxication by enzymes) beyond preexisting
rings and extended chains. Carbon can also mechanisms that operate against natural xeno-
form stable bonds with hydrogen, oxygen, and biotics (see Box 1.1). In this respect, they dif-
nitrogen atoms. fer from inorganic pollutants and from those
Molecules built of carbon alone (e.g., naturally occurring xenobiotics that have sub-
graphite and diamond) or of carbon and hydro- stantial toxicity (e.g., nicotine, pyrethrins, and
gen (hydrocarbons) have very little polarity rotenone are compounds produced by plants
and consequently low water solubility. Polar that are highly toxic to certain species of
molecules have electrical charge associated insect). Aromatic hydrocarbons represent a
with them; nonpolar molecules have little or special case. They have been generated by the
none. Molecules with a strong charge are combustion of organic matter since the appear-
described as highly polar; molecules of low ance of higher plants on earth (e.g., as a result
charge have low polarity. Polar compounds of forest fires started by volcanic lava). Like
tend to be water soluble because the charges metals that are mined, their environmental lev-
on them are attracted to opposite charges on els increase substantially as a consequence of
water molecules. For example, a positive human activity (as with the combustion of coal
charge on an organic molecule will be attached or gasoline to produce aromatic hydrocar-
to a negative charge on a water molecule. Car- bons).
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8 Principles of Ecotoxicology

BOX 1.1 CHEMICAL WARFARE

Chemical warfare has been in progress for a very long period on the evolutionary scale, and the
employment of chemical weapons by humans is, from this point of view, a very recent event.
In higher plants, for example, enzymes have evolved that can synthesise secondary compounds
with high toxicity toward the animals that feed upon them, thereby giving protection against
grazing. In response, detoxication mechanisms (e.g., detoxifying enzymes) have evolved in
mammals; they give protection against these plant toxins. Taken together, these two related
phenomena have been termed a coevolutionary arms race (Ehrlich and Raven, 1964; Harborne,
1993), during the course of which many toxic natural xenobiotics have emerged, together with
defense mechanisms against them. The capacity to synthesise chemical weapons has also evolved
in mammals, both weapons of defense and others that are used for attack. Microorganisms
produce antibiotics that are toxic to other microorganisms that compete with them.
The use of pesticides, biocides, and chemical warfare agents by man should be seen against
this background (see Walker, 2001). Human beings are truly newcomers to this game. Indeed,
early pesticides were frequently natural products. Examples include pyrethrins, nicotine, and
rotenone, all of which have been used as insecticides. Also, the design and subsequent synthesis
of novel pesticides has sometimes been based upon the structure of toxic natural xenobiotics.
Examples include the neonicotinoid insecticides related to nicotine, anticoagulant rodenticides
related to coumarol, pyrethrins related to pyrethrum, and carbamate insecticides related to
physostigmine (see Chapter 1, Section 1.2, and Walker, 2001).
In view of the foregoing, it is hardly surprising that most novel pesticides are readily
degraded by detoxifying enzymes (e.g., monooxygenases and/ or esterases) that were present
in target species long before the discovery of the compounds. It is generally believed that
enzymes of this type have evolved during the course of the coevolutionary arms race.
It is also worth reflecting on the current popular belief in the safety of what is termed
organic produce; in other words, food that is relatively low in synthetic organic chemicals such
as pesticides, pharmaceuticals, and so on. In fact, many natural products are known to be highly
toxic, including botulinum toxin, tetradotoxin, aflatoxin, ergot alkaloids, and many others (see
Trewavas, 2004). All the examples given have been involved in human food poisoning, so the
fact that a product is “organic” does not in itself guarantee safety! At least pesticides and
pharmaceuticals have been subject to rigorous testing and consequent bans and restrictions that
take account of human health risks.

1.2.1 Hydrocarbons Hydrocarbons are divisible into two

classes, (i) alkanes, alkenes, and alkynes, and
(ii) aromatic hydrocarbons (Figure 1.3). The
Hydrocarbons are compounds composed of the distinguishing feature of aromatic hydrocar-
elements carbon and hydrogen only. Some bons is the presence of one or more benzene
hydrocarbons of low molecular weight (e.g., rings in their structure. Benzene rings are six-
methane, ethane, and ethylene) exist as gases membered carbon structures that are unsatur-
at normal temperature and pressure. However, ated in the sense that not all available carbon
most hydrocarbons are liquids or solids. They valences are taken by linkage to hydrogen. In
are of low polarity (i.e., electrical charge, see fact, benzene rings have delocalized electrons
above) and consequently have low water solu- that can move freely over the entire ring system
bility, but they have high solubility in oils and and do not remain in the immediate vicinity of
in most organic solvents. (They are not very any one atom. Other hydrocarbons do not have
soluble in polar organic solvents such as meth- this feature. They vary greatly in molecular size
anol and ethanol.) and may be fully saturated (e.g., hexane and
3635_C001.fm Page 9 Wednesday, November 9, 2005 8:18 AM

Major Classes of Pollutants 9

H H bers of the series are gases, the higher members

H H H H C H liquids or solids.
C C Aromatic hydrocarbons exist as liquids or
H H
H C C C H
H H solids — none of them has a boiling point below
C C
H H H H H
C 80˚C at normal atmospheric pressure. They are
H H more reactive than alkanes, being susceptible
Propane Cyclohexane
to chemical and biochemical transformation.
There are many polycyclic aromatic hydrocar-
bons (PAHs) that are planar (i.e., flat) molecules
consisting of three or more six-membered (ben-
zene) rings directly linked together.
The major sources of hydrocarbons are
deposits of petroleum and natural gas in the
Benzene 3,4-Benzo(a)pyrene upper strata of the Earth’s crust. These fossil
fuels originate from the remains of plants and
Figure 1.3
animals of earlier geological times (notably the
Hydrocarbons are composed of only hydrogen and carbon.
These compounds have low polarity and thus low water Carboniferous period). Although nonaromatic
solubility but high solubility in oils and organic solvents. hydrocarbons predominate in these deposits,
Propane and cyclohexane are examples of alkanes, benzene, crude oils also contain significant amounts of
and benzo(a)pyrene of aromatic compounds. The latter con- PAHs. PAHs are also formed as a consequence
tain six-membered carbon rings (benzene rings) with delo-
of the incomplete combustion of organic mate-
calized electrons. This is indicated by representing the
benzene ring as a hexagon (the six-membered carbon frame) rials. Thus they are generated when coal, oil,
and a circle (the cloud of delocalized electrons) situated or gasoline are burned, when trees or houses
within it. Aromatic hydrocarbons undergo certain character- burn, and when people smoke cigarettes. Major
istic biotransformations influenced by the delocalized elec- sources of hydrocarbon pollution are spillage
trons (Section 5.1.5).
of crude oils (e.g., tanker disasters) and the
combustion of fossil fuels (notably the use of
octane) or unsaturated. Unsaturated hydrocar- brown coal in parts of Eastern Europe).
bons contain carbon–carbon double bonds (e.g.,
ethylene) or carbon–carbon triple bonds (e.g.,
acetylene). Saturated hydrocarbons are referred 1.2.2 Polychlorinated
to as alkanes, unsaturated hydrocarbons with a Biphenyls (PCBs)
carbon–carbon double bond are alkenes, and
unsaturated hydrocarbons with a carbon–car- These are commercial mixtures of related com-
bon triple bond are alkynes. They may exist as pounds (congeners) that are useful for their
single chains, branched chains, or rings (Figure physical properties. They are stable, unreactive
1.3). The properties of these two groups of viscous liquids of low volatility that have been
hydrocarbons will now be considered sepa- used as hydraulic fluids, coolant–insulation flu-
rately. ids in transformers, and plasticizers in paints.
The properties of nonaromatic hydrocar- There are altogether 209 possible PCB conge-
bons depend upon molecular weight and degree ners, and some 120 of these are present in com-
of unsaturation. Alkanes are essentially stable mercial products such as Aroclor 1254, Aroclor
and unreactive and have the general formula 1260, and Clophen A60. The last two digits in
CnH2n + 2. The first four members of the series the numbers refer to the percentage of chlorine
exist as gases (<n = 4). Where n = 5–17, they in the PCB mixture. In either case, the larger
are liquids at normal temperature and pressure. the number, the greater the proportion of higher
Where n = 18 or more, they are solids. Alkenes chlorinated PCBs in the mixture.
and alkynes are more chemically reactive PCB mixtures have very low solubility in
because they contain carbon–carbon double or water but high solubility in oils and organic
triple bonds. As with alkanes, the lower mem- solvents of low polarity. The water solubilities
3635_C001.fm Page 10 Wednesday, November 9, 2005 8:18 AM

10 Principles of Ecotoxicology

Cl
H
γ HCH (Lindane) Cl Cl C Cl p,p’ -DDT
Cl
Cl C
Cl Cl Cl
Cl Cl
Cl
Cl Cl
Cl Cl
Cl
Cl Dieldrin (HEOD)
Aldrin (HHDN) Cl
O
Cl
Cl HH Cl
H H Cl

PCBs Cl Cl Cl Cl
Cl
Cl Cl Cl Cl Cl Cl

Cl
Cl Cl Cl Cl
3,3ʼ,4,4ʼ-Tetrachlorobiphenyl 3,3ʼ,4,4ʼ,5,5ʼ-Hexachlorobiphenyl 2,2ʼ,4,4ʼ,6,6ʼ-Hexachlorobiphenyl
(Coplanar) (Coplanar) (Not coplanar)

Cl O Cl Cl Cl
PCDDs PCDFs

Cl O Cl Cl O Cl
2,3,7,8-tetra-chlorodibenzodioxin 2,3,7,8-tetra-chlorodibenzofuran
(TCDD) (TCDF)

Figure 1.4
Organohalogen compounds are organic compounds containing halogen atoms (the halogen elements are fluorine, chlorine,
bromine, and iodine). All the examples given here are of organochlorine compounds, although it should be noted that
organofluorine compounds (e.g., chlorfluorocarbons) and organobromine compounds (e.g., polybrominated biphenyls) are
also environmental pollutants. The compounds shown here are stable solids of low polarity and water solubility. They are
not found in nature, and in many cases they are only slowly metabolized and consequently are persistent in living organisms
(Chapter 5).

of Aroclor 1254 and Aroclor 1260 are only 21 tries, the use of PCBs is now banned or
μg l–1 and 2.7 μg l–1, respectively. severely restricted.
The individual congeners of PCB vary in Major sources of pollution are (or have
their stereochemistry, depending on the posi- been) manufacturing wastes and the careless
tions of substitution of chlorine atoms. Where disposal or dumping of the liquids referred to
there is no substitution in the ortho positions, above (Waid, 1985–1987).
the two benzene rings tend to remain in the
same plane (coplanar PCBs); 3,3’,4,4’-tetra-
chloro-biphenyl is an example of a coplanar 1.2.3 Polychlorinated
PCB (Figure 1.4). By contrast, substitution of Benzodioxins (PCDDs)
two, three, or four ortho positions with chlorine
leads to the movement of the rings out of plane
because of the interaction of adjacent chlorines The best known member of this group of com-
in different rings (chlorine atoms are bulky). pounds is 2,3,7,8-tetrachlorodibenzodioxin
The molecular conformation is not then a copla- (2,3,7,8-TCDD) (Figure 1.4), usually referred
nar one but is a more globular structure. to simply as dioxin. This is a compound of
PCBs were once used for a number of extremely high toxicity to mammals (LD50
purposes — as dielectric fluids, heat trans- 10–200 μg kg l–1 in rats and mice). In struc-
former fluids, lubricants, and vacuum pump ture, these are flat molecules, formed by the
fluids, as plasticizers (e.g., in paints), and for linking of two benzene rings by two oxygen
making carbonless copy paper. In many coun- bridges with varying substitutions of chlorine
3635_C001.fm Page 11 Wednesday, November 9, 2005 8:18 AM

Major Classes of Pollutants 11

BOX 1.2 PCDDS

The problem of environment pollution by PCCDs is best illustrated by reference to three

well-publicized examples involving 2,3,7,8-TCDD.
1. 2,3,7,8-TCDD (dioxin) can occur as a contaminant of commercial preparations of
herbicides such as 2,4-D and 2,4,5-T. There have been a number of investigations into
pollution caused by these preparations, most notoriously the spraying by the US Air Force
of the Vietnamese jungle with the defoliant herbicide Agent Orange in 1960 to 1969 (see
Section 1.2.11).
2. The release of PCDDs by combustion furnaces used to dispose of PCB wastes, e.g., at the
Rechem plant in Scotland in the 1980s. PCDD residues were detected in soil and cattle in
the surrounding area. The release of these compounds indicated incorrect operation of the
furnace.
3. The release of PCDDs into the air from the Seveso Chemical Works in northern Italy in
1976. The PCDDs were formed in a chemical cloud that contained trichlorophenols.
Some people who had been exposed developed the skin condition chloracne. There were,
however, no fatalities or serious toxic effects attributable to PCDD.

on the available ring positions. There are 75 not, however, received as much attention as
possible congeners of PCDD. PCDDs are PCDDs and do not appear to have raised such
chemically stable compounds with very low serious environmental problems.
water solubilities (less than 1 μg 1–1 at 20˚C)
and limited solubility in most organic sol-
vents, even though they have a lipophilic char- 1.2.5 Polybrominated
acter. Biphenyls (PBBs)
PCDDs are not produced commercially but
are unwanted by-products generated during the Mixtures of polybrominated biphenyls have
synthesis of other compounds. They are also been marketed as fire retardants (e.g., “Firemas-
formed during the combustion of PCBs (fires ter”). These mixtures bear a general resem-
or chemical waste disposal) and by the interac- blance to PCB mixtures and are lipophilic,
tion of chlorophenols during disposal of indus- stable, and unreactive. As with PCBs, some
trial wastes. In general, they are formed when congeners are very persistent in living organ-
chlorophenols interact. isms and have long biological half-lives. In one
PCDD residues have been detected very incident in the USA, a PBB mixture was acci-
widely in the environment (especially in the dentally fed to cattle, leading to the appearance
aquatic environment), albeit at low concentra- of substantial residues in meat products and
tions, e.g., in fish and fish-eating birds (Elliott humans in Wisconsin and neighboring states
et al., 2001). (Carter, 1976).

1.2.4 Polychlorinated 1.2.6 Organochlorine Insecticides

Dibenzofurans (PCDFs)
Organochlorine insecticides are a relatively
These compounds are similar to PCDDs both large group of insecticides with considerable
in structure (Figure 1.4) and in origin. Once diversity of structure, properties, and uses
again, there are many congeners, and the com- (Brooks, 1974). Three major types will be men-
pounds arise as unwanted by-products — they tioned here — DDT and related compounds,
are not synthesized intentionally. They have the chlorinated cyclodiene insecticides (e.g.,
3635_C001.fm Page 12 Wednesday, November 9, 2005 8:18 AM

12 Principles of Ecotoxicology

aldrin and dieldrin), and hexachlorocyclohex- the 1950s), and some of them gave rise to seri-
anes (HCHs) such as lindane (Figure 1.4). ous environmental problems because they (or
Organochlorine insecticides are stable sol- their stable metabolites) have both high toxicity
ids of limited vapor pressure, very low water to vertebrates and marked biological persis-
solubility, and high lipophilicity. Some of them tence. Aldrin, dieldrin, and heptachlor are exam-
are highly persistent in their original form or as ples of cyclodiene insecticides showing this
stable metabolites. All the examples given here undesirable combination of properties (acute
are nerve poisons (see Chapter 7). oral LD50 to rats 40–60 mg kg–1). Chlordane is
Commercial DDT contains 70–80% of the a similar insecticide, but is of lower vertebrate
insecticidal isomer p,p′-DDT. Related insecti- toxicity than the foregoing examples. Endrin
cides include rhothane (DDD) and methoxy- and, to a lesser extent, endosulphan are cyclo-
chlor. The insecticidal properties of DDT were diene insecticides of very high vertebrate toxic-
discovered by Paul Müller of the firm Ciba- ity but only limited biological persistence. In
Geigy in 1939. DDT was used, mainly for vec- general, the cyclodienes resemble DDT in being
tor control, during the Second World War and stable lipophilic solids of very low water solu-
came to be very widely used thereafter for the bility but differ from it in their mode of action
control of agricultural pests, vectors of disease (see Chapter 7). Endosulphan is an exception to
(e.g., malarial mosquitoes), and ectoparasites of this rule, having appreciable water solubility.
farm animals and insects in domestic and indus- The cyclodienes were introduced into
trial premises. Because of its low solubility in Western countries during the 1950s and were
water (<1 mg 1–1), DDT has been formulated used in diverse formulations for many different
as an emulsifiable concentrate for application purposes. Because of their insolubility in water,
as a spray. (Emulsifiable concentrates are solu- emulsifiable concentrates and wettable powders
tions of pesticides in organic liquids; when were the formulations normally used for spray-
added to water, they form creamy emulsions ing. Sprays were used for the control of certain
that can be sprayed on crops.) crop pests and for certain vectors of disease
DDT has an acute oral LD50 of 113–450 (e.g., tsetse fly). They were also used in dips
mg kg–1 and is considered to be only moderately and sprays to control ectoparasites of livestock
toxic to vertebrates (Chapter 6). However, it has and were very widely used as seed dressings
been shown to cause eggshell thinning in cer- for cereals and other crops. The use of aldrin,
tain sensitive species of birds at very low doses dieldrin, and heptachlor for the latter purpose
through the action of its stable metabolite p,p′- had very serious ecological consequences,
DDE (see Chapters 7, 12, and 16). which will be discussed in some detail in Chap-
In addition to p,p′-DDT, some 20% of com- ters 12 and 16.
mercial insecticide is represented as o,p′-DDT, By the 1990s, few uses of either DDT and
which is more readily biodegradable than p,p′- its relatives or of the cyclodiene insecticides
DDT and has very low toxicity to insects and remained. The use of these compounds for most
vertebrates. On the other hand, it has been purposes had been banned on the ground of
shown to have estrogenic activity, for example perceived human health risks or hazards to the
in rats, and has sometimes, together with other environment. However, some of these com-
compounds of the DDT group, been implicated pounds continue to be used on a limited scale
in cases of alleged endocrine disruption in the in some countries. For example, some use is
natural environment (see Chapter 7). still made of DDT in certain developing coun-
Kelthane (dicofol) is another pesticide tries to control vectors such as the malarial mos-
related in structure to DDT, which has been quito, which is seen to represent a greater
marketed as an acaricide. It has only weak hazard to people than the side effects of the
insecticidal activity, is of limited persistence, chemical. Although these compounds are little
and there is some evidence that it may act as used today, their ecotoxicology is discussed in
an endocrine disruptor (see Chapter 7). some detail in parts of the ensuing text for two
The chlorinated cyclodiene insecticides reasons. On the one hand, the very marked per-
were introduced after DDT (very widely during sistence of compounds such as dieldrin and
3635_C001.fm Page 13 Wednesday, November 9, 2005 8:18 AM

Major Classes of Pollutants 13

p,p′-DDE has ensured that significant residues 1.2.7 Organophosphorous

are still present in once heavily contaminated Insecticides (OPs)
soils and/or sediments and will only slowly dis-
appear over the decades to come. These resi- During the Second World War, interest devel-
dues are still slowly released into aquatic and oped in organophosphorous compounds, which
terrestrial food chains and can reach significant act as nerve poisons (neurotoxins) because of
concentrations in animals at higher trophic lev- their ability to inhibit the enzyme acetylcho-
els. The second reason is that they have been linesterase (AChE) (Chapter 7). These com-
studied in considerable depth and detail — pounds were produced for two main uses — as
probably more so than any other type of organic insecticides and as chemical warfare agents
pollutant. A great deal has been learned about (nerve gases) (Ballantyne and Marrs, 1992).
the ecological hazards associated with them, They are organic esters of phosphorus acids
and they serve as useful examples and models (Figure 1.5). Today, a large number of organo-
for persistent lipophilic pollutants more gener- phosphorous compounds are marketed as insec-
ally in the present text. Further, their persistence ticides, and nearly all of them correspond to the
is associated with a potential to cause neuro- basic formula shown in Figure 1.5.
toxic effects and associated behavioral effects Most organophosphorous insecticides
at sublethal levels (see 15.13, Example B). (OPs) are liquids of lipophilic character and
Hexachlorocyclohexane (HCH) has been some volatility; a few are solids. They are, in
marketed as a crude mixture of isomers (“BHC”) general, less stable than organochlorine insec-
but more extensively as a refined product con- ticides and are more readily broken down by
taining mainly the γ isomer, known as γ-HCH, chemical or biochemical agencies (Eto, 1974;
γ-BHC, or lindane (Figure 1.4). γ-HCH has sim- Fest and Schmidt, 1982). Thus they tend to be
ilar properties to other organochlorine insecti- relatively short-lived when free in the environ-
cides but is somewhat more polar and water ment, and the environmental hazards that they
soluble (7 mg l–1). Emulsifiable concentrates of present are largely, but not exclusively, associ-
HCH have been used for controlling agricultural ated with short-term (acute) toxicity. There is
pests or parasites of farm animals. It has also concern, as with other neurotoxic insecicides,
been used as an insecticidal seed dressing (e.g., about their behavioral effects upon animals at
on cereal seed). HCH is only moderately toxic sublethal doses. They are more polar and water
to rats (LD50 60–250 mg kg–1). soluble than the main types of organochlorine

Organophosphorus insecticides: general formula

S

C2H5O P O
RO O or S
C2H5O N CH3
P R = Alkyl group
X = Leaving group CH
Diazinon CH3
RO X N
CH3

Carbamate insecticides: general formula H

O C N
O CH3 CH3
O
X O C N (1-Naphthyl-
N- methylcarbamate)
H or CH3
X = Organic group
(e.g. aromatic or heterocyclic) Carbaryl

Figure 1.5
Organophosphorous and carbamate insecticides. These compounds are toxic to insects because they inhibit the enzyme
acetylcholinesterase (Chapter 7). They vary in their polarity and water solubility. They are generally more reactive and less
stable and persistent than the organochlorine insecticides. The leaving group of the organophosphorous compounds breaks
away from the rest of the molecule when hydrolysis occurs (Chapter 5).
3635_C001.fm Page 14 Wednesday, November 9, 2005 8:18 AM

14 Principles of Ecotoxicology

insecticides. Their water solubility is highly cides for controlling slugs and snails. It is impor-
variable, with some compounds (e.g., tant to distinguish between the insecticidal
dimethoate) having appreciable solubility. The carbamates and herbicidal carbamates (e.g.,
active forms of some OPs have sufficient water propham, chlorpropham), which have only low
solubility to be effective systemic insecticides, toxicity to animals.
reaching high enough concentrations in the Carbamate insecticides are formulated in a
phloem of plants to poison sap-feeding insects similar way to OPs, with the most toxic ones
(cf. organochlorine compounds and pyre- (e.g., aldicarb and carbofuran) being available
throids). only as granules. They are used, principally, to
The formulation of organophosphorous control insect pests on agricultural and horti-
compounds is important in determining the cultural crops, although they also have some
environmental hazards that they present. Many use for control of nematodes (i.e., as nemati-
are formulated as emulsifiable concentrates for cides) and molluscs (i.e., as molluscicides).
spraying. Others are incorporated into seed
dressings or into granular formulations. Gran-
ular formulations are required for the most toxic 1.2.9 Pyrethroid Insecticides
OPs (e.g., disyston and phorate) because they
are safer to handle than emulsifiable concen- Naturally occurring pyrethrin insecticides,
trates or certain other types of formulation. The which are found in the flowering heads of Chry-
insecticide is locked up within the granule and santhemum spp., provided the model for the
is only slowly released into the environment. development of synthetic pyrethroids. Synthetic
In many countries, OPs are still applied to pyrethroids are, in general, more stable chemi-
crops as sprays, granules, seed dressings and cally and biochemically than are natural pyre-
root dips. They are used to control ectoparasites thrins (Leahy, 1985). Pyrethroids are solids of
of farm and domestic animals (commonly in very low water solubility that act as neurotoxins
sheep dipping) and sometimes also for control- in a way similar to DDT (see Chapter 7). They
ling internal parasites (e.g., the ox warble fly). are esters formed between an organic acid (usu-
Other uses include control of certain vertebrate ally chrysanthemic acid) and an organic base
pests (e.g., the bird Quelea in parts of Africa), (see Figure 1.6). Although pyrethroids are more
locusts, stored product pests, especially beetles, stable than pyrethrins, they are readily biode-
insect vectors of disease such as mosquitoes gradable and do not have long biological half-
and parasites of salmon at fish farms (Hites et lives. They can, however, bind to particles in
al., 2003). soils and sediments and show some persistence
in these locations. With their low water solubil-
ities, they do not show significant systemic
1.2.8 Carbamate Insecticides properties and are not used as systemic insec-
ticides. The hazards that they present relate
Carbamate insecticides are derivatives of car- mainly to short-term toxicity. However, it
bamic acid that have been developed more should be emphasized that they are highly
recently than organochlorine compounds (OCs)
and OPs (Figure 1.5) (Kuhr and Dorough, 1977).
CH3 CH3
Like OPs, however, they act as inhibitors of ace-
O
tylcholinesterase. Carbamates are frequently sol- Cl
C CH O
ids, sometimes liquids. They vary greatly in C CH2
Cl
water solubility. Like OPs, they are readily
O
degradable by chemical and biochemical agen-
cies and do not usually raise problems of persis- Permethrin

tence. The main hazards that they present relate

Figure 1.6
to short term toxicity. Some of them (e.g., aldi- Pyrethroid insecticides. These have low polarity and limited
carb and carbofuran) act as systemic insecticides. water solubility. They are related in structure to the natural
A few (e.g., methiocarb) are used as mollusci- pyrethrins, which are also toxic to insects.
3635_C001.fm Page 15 Wednesday, November 9, 2005 8:18 AM

Major Classes of Pollutants 15

NO2 R
N 2’ 1’
Cl O CH COOH
N N H CH3

N R = CH3 or Cl
CI
Phenoxyacetic acids (general formula)
FIGURE 1.7
Imidacloprid is an example of a neonicotinoid insecticide,
Figure 1.8
which shows a structural resemblance to nicotine.
Phenoxyalkanoic acid herbicides. The example given is a
general formula for phenoxyacetic acids, which include
selective between insects on the one hand and 2,4-D and MCPA. There are also phenoxypropionic acids
mammals and birds on the other. The main envi- (e.g., CMPP) and phenoxybutyric acids (e.g., 2,4-DB). All
of them have plant-growth-regulating properties and have
ronmental concerns relate to their toxicity to some resemblance to the natural growth regulator
fish and nontarget invertebrates.There is also indoleacetic acid.
concern about their effects upon animal behav-
ior at sublethal doses (8.4.2).
Pyrethroids are formulated mainly as emul- ples are 2,4-D, MCPA, CMPP, 2,4-DB, and
sifiable concentrates for spraying. They are 2,4,5-T (for general formulae, see Figure 1.8).
used to control a wide range of insect pests of They act by disturbing growth processes in a
agricultural and horticultural crops throughout manner akin to that of the natural plant growth
the world and are coming to be used extensively regulate indoleacetic acid. They are deriva-
to control insect vectors of disease (e.g., the tives of phenoxyalkane carboxylic acids.
tsetse fly in parts of Africa). When formulated as alkali salts, they are
highly water soluble; when formulated as sim-
ple esters, they are lipophilic and of low water
1.2.10 Neonicotinoids solubility.
Most phenoxy herbicides are readily bio-
Another group of neurotoxic insecticides intro- degradable and so are not strongly persistent in
duced since the pyrethroids became widely living organisms or in soil. They are selective,
available are the neonicotinoids. These are i.e., selectively toxic between monocotyledon-
structurally similar to the natural compound ous and dicotyledonous plants. Their principal
nicotine. Like nicotine, they can interact with use is to control dicot weeds in monocot crops
nicotinic receptors located on cholinergic syn- (e.g., cereals and grasses). The environmental
apses of animals. However, they are less polar hazards are of two kinds. First, there is the
than the natural compound and show more tox- problem of unwanted phytotoxicity as a conse-
icity toward insects than to vertebrates. An quence of spray or vapor drift. Second, formu-
example of a neonicotinoid is imidacloprid, the lations of certain herbicides of this type have
structure of which is shown in Figure 1.7. As sometimes been contaminated with the highly
with other neurotoxic insecticides, there is evi- toxic compound TCDD, e.g., Agent Orange, a
dence that sublethal doses can cause behavioral formulation containing 2,4-D and 2,4,5-T that
effects (See 8.4.2 ). was used as a defoliant in Vietnam (Stellman
et al., 2003) (see Section 1.2.3).
Water-soluble salts (e.g., Na+, K+) are for-
1.2.11 Phenoxy Herbicides mulated as aqueous solutions (aqueous concen-
(Plant-Growth-Regulator trates), whereas lipophilic esters are formulated
Herbicides) as emulsifiable concentrates. The latter have
sometimes caused unwanted phytotoxicity
Phenoxy herbicides constitute the single most owing to the volatility of some of the esters
important group of herbicides. Familiar exam- (vapor drift).
3635_C001.fm Page 16 Wednesday, November 9, 2005 8:18 AM

16 Principles of Ecotoxicology

OH nonpolar (hydrophobic) C–C chains. Nonionic

H
detergents have no such permanent charge;
C rather they have a number of atoms that are
weakly electropositive and electronegative.
CH2
O O This is due, in the examples shown, to the elec-
COCH3 tron-attracting power of oxygen atoms.
Detergents are very widely used in both
Figure 1.9 domestic and industrial premises. The major
Rodenticides. Warfarin and related compounds, such as entry point into water is via sewage works into
diphenacoum and brodifacoum, are anticoagulant rodenti-
cides. They are complex molecules bearing some structural
surface waters. They are also used in pesticide
resemblance to vitamin K. Their toxic action is due to formulations and for dispersing oil spills at
competition with vitamin K in the liver (vitamin K antag- sea.
onism). The degradation of alkylphenol polyethox-
ylates (nonionic detergents) can lead to the for-
1.2.12 Anticoagulant m a t i o n o f a l ky l p h e n o l s ( p a r t i c u l a r l y
Rodenticides nonylphenols), which act as endocrine disrup-
tors (see Chapter 10).
For many years, the compound warfarin (Figure
1.9) has been used as a rodenticide. It is a lipo-
philic molecule of low water solubility that acts 1.2.14 Chlorophenols
as an antagonist to vitamin K (Chapter 7). More
recently, as wild rodents have developed resis- A number of different polychlorinated phenols
tance to warfarin, a number of second-genera- (PCPs) occur as environmental pollutants
tion anticoagulant rodenticides (sometimes (Figure 1.11). A major source is pulp mill
called super warfarins) have been marketed, effluent, from which they arise because of the
which are structurally related to warfarin. These chemical action of chlorine (used as a bleach-
include diphenacoum, bromadiolone, brodifa- ing agent) upon phenolic substances present
coum, and flocoumafen and resemble warfarin in wood pulp (Södergren, 1991). Pentachlo-
in their general properties but are more toxic to rophenol is used as a wood preservative and
mammals and birds and are markedly persistent this is an important source of pollution. Chlo-
in the livers of vertebrates. Thus they may be rinated phenols have acidic properties and are
transferred from rodents to the vertebrate pred- water soluble, chemically reactive, and of lim-
ators and scavengers that feed upon them. Owls, ited persistence. The tendency of some PCPs
for example, have been found to contain resi- to interact and form PCDDs was discussed in
dues of them in the UK. Rodenticides are usu- Section 1.2.3.
ally incorporated into bait, which is then placed
in buildings or out of doors, where it will be
taken by wild rodents. 1.2.15 Ethinylestradiol (EE2)

EE2 is a potent estrogen , widely used in oral

1.2.13 Detergents contraceptives. It has a close structural resem-
blance to the natural hormone estradiol (Fig-
Detergents are organic compounds that have ure 1.12). It shows some persistence in fish
both polar and nonpolar characteristics. They and in surface waters. Its persistence in fish is
tend to exist at phase boundaries, where they partly due to the tendency of its glucuronide
are associated with both polar and nonpolar conjugate to undergo enterohepatic circula-
media. Some examples are shown in Figure tion, as the result of the action of the enzyme
1.10. Detergents are of three types, anionic, B-glucuronidase in the gut (see Section 5.1.6).
cationic, and nonionic. The first two have per- The glucuronide seems to be relatively unsta-
manent negative or positive charges attached to ble in the environment. There is concern that
3635_C001.fm Page 17 Wednesday, November 9, 2005 8:18 AM

Major Classes of Pollutants 17

Anionic
H H

H H H HH C H H HH C H O

H C C C C C C C C S O-Na+

H H H HH C H H HH C H O
H H
Sodium tetrapropylene
benzene sulphonate (hard) H H
H H H H H H H H HH C H O

H C C C C C C C C C C S O-Na+

H H H H H H H H HH C H O

Dobane J.N. sulphonate H H

(soft)
O

CH3(CH2)n O S O-Na+ Sodium alkyl sulphonate (soft)

Cationic H H
H C (CH2)14 C N+ Br-

H H

Cetyl pyridinum bromide

Nonionic

Polyglycol ethers of alkylated HO (C2H4O)n R R = alkyl group

phenols, e.g., lissapol N stergene

Figure 1.10
Detergents are molecules that have both polar and nonpolar elements. They may have permanent negative charge (anionic
detergents), permanent positive charge (cationic detergents), or a collection of small positive and negative charges over
their structure (nonionic detergents).

OH
Cl
OH
CH3 C CH
Cl
H
Cl

H H
2,4,5-Trichlorophenol
HO
Figure 1.11
Figure 1.12
Chlorinated phenols have acidic properties, releasing H+
17 A- Ethinylestradiol (EE2). A potent synthetic estrogen
ions when they dissolve in water. They can interact to form
bearing a structural resemblance to estradiol.
dioxin (see Section 1.2.3).
3635_C001.fm Page 18 Wednesday, November 9, 2005 8:18 AM

18 Principles of Ecotoxicology

EE2, acting in concert with other environmen- is demonstrated by its use as a lining in food
tal estrogens present in surface water, may containers.
cause the feminization and consequent infer- Nevertheless, the toxicity of several met-
tility of male fish. The possible ecological con- als is greatly enhanced if they become bound
sequences of this are discussed in Section either deliberately or accidentally to an
15.2. organic ligand. This changes their chemical
behavior in the environment and within organ-
isms. Metals are modified in this way to
1.2.16 Pharmaceuticals increase their toxicity for use as pesticides.
Organomercury compounds were used widely
With recent rapid advances in medical science as antifungal seed dressings in the UK until as
and the associated growth of the pharmaceu- recently as 1993. Organolead compounds have
tical industry, it is not surprising that traces of been used extensively for control of caterpil-
drugs are sometimes detected in surface lars on fruit crops. Organotin compounds, par-
waters. Where drugs are widely used they are ticularly tributyl tin, are extremely toxic. Their
likely to appear as contaminants of sewage main use is for preserving timbers from the
outflows. Indeed, the case of EE2 discussed activities of aquatic boring animals and as a
above is an example of this. On the available component of antifouling paints that are
evidence, levels of drugs detected in surface applied to the outer surface of boats and fish
waters appear relatively low and unlikely to cages to inhibit settlement by marine organ-
cause ecotoxicity. For example, a study of the isms. When these substances leach into the
beta blockers propranolol, metoprol and nad- environment, they can affect nontarget organ-
olol suggested that environmental levels were isms. Tributyl tin, for example, has devastated
not high enough to have significant toxic populations of the dog whelk Nucella lapillus
effects on aquatic invertebrates (Huggett et al., near sites of boating activity in many countries
2002). However, little work has been done in (see Sections 13.6.4 and 16.3).
this field as yet, and there are no grounds for A tragic example of the effects of organo-
complacency. With so many compounds now mercury compounds occurred in Minamata
there is a question of possible potentiation of Bay, Japan, in the 1950s. Metallic mercury
toxicity (see Chapter 9) with the complex mix- released from a paper factory on the shores of
tures of environmental chemicals that now the bay was methylated in the sediments by
occur in some surface waters, albeit at very bacteria to form methyl mercury (see Kudo et
low concentrations. There are also questions al., 1980). Mercury in its methylated form is
about pollution arising from farms where anti- much more bioavailable than liquid mercury
biotics and drugs used to treat farm animals and it passed rapidly along the food chain until
or fish may contribute to the pollution of fresh it reached high concentrations in fish. The
waters. local people relied heavily on locally caught
fish and were thus vulnerable to poisoning.
About 100 people died and many suffered
severe disabilities from mercury poisoning.
1.3 Organometallic Such incidents are most severe when a local
Compounds population is highly dependent on a single
food source but are rare in more developed
Some metal ions are so insoluble that they are regions where food is obtained from much
relatively nontoxic to animals if ingested. Liq- wider sources. Similar problems occur in the
uid metal mercury, for example, can be swal- Amazon Basin. Huge quantities of mercury
lowed in small amounts by humans with little are being dumped into the river as a by-prod-
long-term effect. Indeed, until the last century, uct of gold refining, and there is evidence that
drinking liquid mercury was recommended as this is becoming methylated and passed into
a cure for constipation! The low toxicity of tin food chains (Pfeiffer et al., 1989/1993).
Another Random Scribd Document
with Unrelated Content
 Si looked, and saw Mrs. Bolster striding toward them. Shorty hung
back instinctively for an instant, and then braced up and bade her
good morning.
She grunted an acknowledgment, and said rather imperiously:
"Y're a-gwine, air yo'?"
"Certainly," answered Shorty.
"And yo'?" she inquired, looking at Si.
"He's a-goin', too," answered Shorty. "Mustn't expect him to talk.
He's short on tongue this mornin'. Ketched a bad cold night before
last. Settled on his word-mill. Unjinted his clapper. Can't speak a
word. Doctor says it will last several days. Not a great affliction.
Couldn't 've lost anything o' less account."
"Must've bin an orful cold," said she, taking her pipe from her
mouth and eyeing Si suspiciously.
"Never knowed a cold to shut off any one's gab afore. Seems t' me
that hit makes people talk more. But these Yankees air different.
Whar air yer things? Did yo' bring plenty o' coffee?'
"We've got 'em hid down here in the brush," said Shorty. "We'll git
'em when we're ready to start."
"We're ready now," she answered. "Come along."
"But we hain't no passes," objected Shorty. "We must go to the
Captain and git passes."
"Yo' won't need no passes," she said impatiently. "Foller me."
Shorty had expected to make the pretext about the passes serve
for informing Capt. McGillicuddy of the presence of the woman in the
camp. He looked quickly around and saw the Captain sauntering
carelessly at a little distance, so that any notification was
unnecessary. He turned and followed Mrs. Bolster's long strides, with
Si bringing up the rear.
They went to the clump of brush where they had hidden their
haversacks and guns. Mrs. Bolster eagerly examined the precious
package of coffee.
 "I'll take keer o' this myself," she said, stowing it away about her
lanky person. "I can't afford to take no resks as to hit."
Si and Shorty had thought themselves very familiar with the
campground, but they were astonished to find themselves led
outside the line without passing under the eye of a single guard. Si
looked at Shorty in amazement, and Shorty remarked:
"Well, I'll be durned."
The woman noticed and understood. "Yo' Yanks," she said
scornfully, "think yourselves moughty smart with all your book-larnin',
and yo'uns put on heaps o' airs over po' folks what hain't no
eddication; but what you don't know about Tennessee woods would
make a bigger book than ever was printed."
"I believe you," said Shorty fervently. His superstition in regard to
her was rapidly augmenting to that point where he believed her
capable of anything. He was alarmed a'bout Capt. McGillicuddy's
being able to follow their mysterious movements. But they soon
came to the road, and looking back from the top of a hill, Shorty's
heart lightened as he saw a squad moving out which he was
confident was led by Capt. McGillicuddy.
But little had been said so far. At a turn of the road they came
upon a gray-bearded man, wearing a battered silk hat and
spectacles, whom Mrs. Bolster greeted as "'Squire."
The word seemed to send all the blood from Shorty's face, and he
looked appealingly to Si as if the crisis had come.
The newcomer looked them over sharply and inquired:
"Who are these men, Mrs. Bolster?"
"They'uns 's all right. They'uns 's had enough o' Abolition doin's,
and hev come over whar they'uns allers rayly belonged. This one is a
partickler friend o' mine," and she leered at Shorty in a way that
made his blood run cold.
"Hain't yo' time t' stop a minute, 'Squire?" she asked appealingly,
as the newcomer turned his horse's head to renew his journey.
"Not now; not now," answered the 'Squire, digging his heels into
his steed's side. "I want to talk t' yo' and these 'ere men 'bout what's
gwine on in the Lincoln camps, but I must hurry on now to meet
Capt. Solomon at the Winding Blades. I'll come over to your house
this evening," he called back.
"Don't fail, 'Squire," she answered, "fur I've got a little job for yo',
an' I want hit partickerly done this very evenin'. Hit can't wait."
"I'll be there without fail," he assured her.
"Capt. Solomon's the man what sent the letter to you," she
explained, which somewhat raised Shorty's depressed heart, for he
began to have hopes that Rosenbaum might rescue him if Capt.
McGillicuddy should be behind time.
As they jogged onward farther from camp Mrs. Bolster's saturnine
earnestness began to be succeeded by what were intended to be
demonstrations of playful affection for her future husband, whom she
now began to regard as securely hers. She would draw Shorty into
the path a little ahead of Si, and walk alongside of him, pinching his
arm and jabbering incoherent words which were meant for terms of
endearment. When the narrowness of the road made them walk in
single file she would come up from time to time alongside with cuffs
intended for playful love-taps.
At each of these Shorty would cast such a look of wretchedness at
Si that the latter had difficulty in preserving his steadfast silence and
rigidity of countenance.
But the woman's chief affection seemed to be called forth by the
package of coffee. She would stop in the midst of any demonstration
to pull out the bag containing the fragrant berry, and lovingly inhale
its odor.
It was long past noon when she announced: "Thar's my house
right ahead." She followed this up with a ringing whoopee, which
made the tumbledown cabin suddenly swarm with animation. A
legion of loud-mouthed dogs charged down toward the road.
Children of various ages, but of no variety in their rags and unkempt
wildness, followed the dogs, or perched upon the fence-corners and
stumps, and three or four shambling, evil-faced mountaineers lunged
forward, guns in hand, with eyes fiercer than the dogs, as they
looked over the two armed soldiers.
"They'uns is all right, boys," exclaimed the woman. "They'uns 's
plum sick o' doggin' hit for Abe Lincoln an' quit."
"Let 'em gin up thar guns, then," said the foremost man, who had
but one eye, reaching for Shorty's musket. "I'll take this one. I've
been longin' for a good Yankee gun for a plum month to reach them
Yankee pickets on Duck River."
Though Shorty and Si had schooled themselves in the part they
were to play, the repugnant thought of giving up their arms to the
rebels threatened to overset everything. Instinctively they threw up
their guns to knock over the impudent guerrillas. The woman strode
between them and the others, and caught hold of their muskets.
"Don't be fools. Let 'em have your guns," she said, and she caught
Si's with such quick unexpectedness that she wrenched it from his
grasp and flung it to the man who wanted Shorty's. She threw one
arm around Shorty's neck, with a hug so muscular that his breath
failed, and she wrenched his gun away. She kept this in her hand,
however.
"Now, I want these 'ere men treated right," she announced to the
others, "and I'm a-gwine to have 'em treated right, or I'll bust
somebody's skillet. They'uns is my takings, and I'm a-gwine to have
all the say 'bout 'em. I've never interfered with any Yankees any o'
yo'uns have brung in. Yo've done with them as you pleased, an' I'm
a-gwine to do with these jest as I please, and yo'uns that don't like
hit kin jest lump hit, that's all."
 "'Frony Bolster, I want yo' to take yo'r arms from around that
Yank's neck," said the man who had tried to take Shorty's gun. "I
won't 'low yo' to put yo'r arm 'round another man's neck as long's
I'm alive to stop it."
"Ye won't, Jeff Hackberry," she sneered. "Jealous, air ye? You've
got no bizniss o' bein'. Done tole ye 'long ago I'd never marry yo', so
long as I could find a man who has two good eyes and a 'spectable
character. I've done found him. Here he is, and 'Squire Corson 'll
splice us to-night."
How much of each of the emotions of jealousy, disappointment,
hurt vanity, and rebel antagonism went into the howl that Mr. Jeff
Hackberry set up at this announcement will never be known. He
made a rush with clenched fists at Shorty.
A better description could be given of the operations of the center
of a tornado than of the events of the next few minutes. Shorty and
Hackberry grappled fiercely. Mrs. Bolster mixed in to stop the fight
and save Shorty. Si and the other three rebels flung themselves into
the whirlpool of strikes, kicks, and grapples. The delighted children
came rushing in, and eagerly joined the fray, striking with charming
impartiality at every opportunity to get a lick in anywhere on
anybody; and finally the legion of dogs, to whom such scenes
seemed familiar and gladsome, rushed in with an ear-splitting clamor,
and jumped and bit at the arms and legs that went flying around.
This was too violent to last long. Everybody and everything had to
stop from sheer exhaustion. But when the stop came Mrs. Bolster
was sitting on the prostrate form of Jeff Hackberry. The others were
disentangling themselves from one another, the children and the
dogs, and apparently trying to get themselves into relation with the
points of the compass and understand what had been happening.
"Have yo' had enough, Jeff Hackberry," inquired Mrs. Bolster, "or
will yo' obleege me to gouge yer other eye out afore yo' come to yer
senses?"
"Le' me up, 'Frony," pleaded the man, "an' then we kin talk this
thing over."
 CHAPTER XV. SHORTY NEARLY GOT
MARRIED
BREAKING UP A BAD REBEL NEST IS NO
PICNIC.
WHEN physical exhaustion called a halt in the fracas, Mrs. Bolster
was seated on Jeff Hackberry's breast with her sinewy hands
clutching his long hair, and her thumb, with a cruel, long nail,
pressing the ball of his one good eye. Shorty was holding down one
of the guerrillas who had tried to climb on his back when he was
grappling with Hackberry. Si had knocked one guerrilla senseless with
his gun-barrel, and now came to a breathless standstill in a struggle
with another for the possession of his gun. The children and dogs
had broken up into several smaller stormcenters, in each of which a
vicious fight was going on. In some it was dog and dog; in some
child and child, and in others dogs and children mixed.
Then they all halted to observe the outcome of the discussion
between Mrs. Bolster and Jeff Hackberry.
"Holler 'nuff, Jeff, or out goes yer last light," commanded Mrs.
Bolster, emphasizing her words by rising a little, and then settling
down on Jeff's breast with a force that drove near every spoonful of
breath out of him.
"'Frony, le' me up," he begged in gasps.
"Mrs. Bolster," she reminded him, with another jounce upon his
chest.
"Mrs. Bolster, le' me up. I'd 'a' got away with that 'ere Yank ef ye'
hedn't tripped me with them long legs o' your'n."
"I'm right smart on the trip, aint I," she grinned. "I never seed a
man yit that I couldn't throw in any sort of a rastle."
 "Le' me up, Mrs. Bolster, an le's begin over agin, an' yo' keep out,"
begged Hackberry.
"Not much I won't. I ain't that kind of a chicken," she asserted
with another jounce. "When I down a man I down him fer good, an'
he never gits up agin 'till he caves entirely. If I let yo' up, will yo'
swar to quite down peaceable as a lamb, an' make the rest do the
same?"
"Never," asserted Hackberry. "I'm ergwine to have it out with that
Yank."
"No you haint," she replied with a still more emphatic jounce that
made Hackberry use all the breath left him to groan.
"I'll quit," he said, with his next instalment of atmosphere.
"Will yo' agree t' let me marry this Yank, an' t' give me away as my
oldest friend, nearest o' kin, an' best man?" she inquired, rising
sufficiently to let him take in a full breath and give a free, unforced
answer.
"Nary a time," he shrieked. "I'll die fust, afore I'll 'low yo' t' marry
ary other man but me."
"Then you'll lose yor bClinker, yo' pigheaded, likker-guzzling',
ornery, no-account sand-hill crane," she said, viciously coming down
on his chest with her full weight and sticking the point of hei nail
against his eye. "I wouldn't marry yo' if ye wuz the last nubbin' in the
Lord A'mighty's crib, and thar'd never be another crap o' men. Ye'll
never git no chance to make me yer slave, and beat me and starve
me t' death as yo' did Nance Brill. I ain't gwine t' fool with yer
pervarsity nary a minnit longer. Say this instant whether yo'll do as I
say with a freewill and good heart, or out goes yer peeper." "I
promise," groaned Jeff.
"Yo' sw'ar hit?" she demanded.
"Yes, I sw'ar hit," answered Jeff.
Mrs. Bolster rose, and confirmed the contract by giving him a kick
in the side with her heavy brogan.
"That's jest a lovetap," she remarked, "'t let yo' know t' le' me
alone hereafter. Now, le's straighten things around here fer a
pleasant time."
She initiated her proposed era of good feeling by a sounding kick
in the ribs of the most obstreperous of the dogs, and a slap on the
face of a 12-year-old girl, who was the noisest and most pugnacious
of the lot. Each of these set up a howl, but there was a general
acquiescence in her assertion of authority.

Jeff Hackberry sat up, scratched and rubbed himself, seemed to be

trying to once more get a full supply of air in his lungs, and turned a
one-eyed glare on his surroundings. The guerrilla whom Si had
knocked down began to show signs of returning consciousness, but
no one paid any attention to him. One of the other two pulled out a
piece of tobacco, split it in two, put the bigger half in his mouth and
handed the remainder to his partner. Both began chewing
meditatively and looking with vacant eyes for the next act in the
drama. Shorty regained his gun, and he and Si looked inquiringly at
one another and the mistress of the ranch.
"Come on up t' the house," she said, starting in that direction. The
rest followed, with Si and Shorty in the lead.
The boys gazed around them with strong curiosity. The interior
was like that of the other log cabins they had seen—a rough
puncheon floor for the single room, a fireplace as big as a barn door,
built of rough stones, with a hearth of undressed flat stones, upon
which sat a few clumsy cooking utensils of heavy cast-iron, three-
legged stools for chairs, a table of rough whip-sawed boards held
together by wooden pins. In two of the corners were beds made of a
layer of poles resting upon a stick supported at one end upon a log in
the wall and at the other end a forked stick driven between the
puncheons into the ground below. Upon this was a pile of beech
leaves doing duty as a mattress. The bed-clothes were a mass of
ragged fabrics, sheepskins, etc., used in the daytime for saddle-
blankets and at night upon the bed. There had been added to them,
however, looking particularly good and rich in contrast with their
squalor, several blankets with "U. S." marked upon them. Around the
room were canteens, shoes, and other soldier belongings.
"Have they killed and robbed the men to whom these belonged, or
merely traded whisky for them?" was the thought that instantly
flashed through Si's and Shorty's minds. The answer seemed to be
favorable to murder and robbery. "Set down an' make yourselves at
home. I'll git yo' out suthin' t' wet yer whistles," said Mrs. Bolster,
wreathing as much graciousness as she could into her weathered-
wood countenance. She apparently kicked at the same instant a stool
toward them with her left foot, and a dog out of the way with her
right, a performance that excited Shorty's admiration.
 "When I see a woman kick in different directions with both feet at
the same time, I understood how dangerous her trip would be in a
rastle," he said afterward.
Si and Shorty shoved two of the stools so that they could sit with
their backs to the wall, still holding their guns.
The guerrillas came filing in, with an expectant look on their faces.
Even Jeff Hackberry looked more thirstily longing than wrathful. The
man who had fallen under Si's gunbarrel had gotten able to walk,
was rubbing his head and moaning with the design of attracting
attention and sympathy.
Mrs. Bolster produced a key from her pocket. The others
understood what this meant. They lifted aside some sacks of meal
and shelled corn, and revealed a puncheon which had been cut in
two, and the short piece was garnished by rude iron hinges and
hasp, all probably taken from some burned barn. The hasp was
locked into the staple by one of the heavy padlocks customary on the
plantations, and this Mr. Bolster proceeded to open with her key.
When the puncheon was turned up it revealed a pit beneath, from
which she lifted a large jug of whisky. She poured some out in a tin
cup and handed it to Shorty.
"Take a big swig," she said; "hit's mouty good stuff—ole Jeff
Thompson's brewin' from yaller corn raised on rich bottom land."
Si trembled as he saw his partner take the cup. Shorty smelled it
appreciatively. "That is good stuff," he said. "Roses ain't nowhere
alongside."
He put the cup to his lips and took a sip.
"Tastes as good as it smells," he said, heartily, while the mouths of
the guerrillas were watering. He put the cup again to his lips, as if to
take a deep draft. Then came a short cough and a tremendous
sputter, followed by more painful coughing and strangling.
"Jest my infernal luck," gasped Shorty. "I would talk, an' I got
some down the wrong way.
"Lord, it's burnin' my lights out. Gi' me a drink o' water, somebody."
 One of the children handed him a gourdful of water, while he
continued to cough and sputter and blame himself for talking when
he was drinking.
The woman handed the cup to Si, who feared that the liquor might
be poisoned or drugged. He made a pretense of drinking, and then
handed the cup back, making motions that his throat was so sore
that he could not drink much. Mrs. Bolster looked at him suspiciously,
but the clamor of the guerillas distracted her attention, and she
turned to supply them.
"No, Jeff Hackberry," she said firmly, "yo' can't have more'n two
fingers. I know yo' of old, an' jest how much yo' orter tote. Two
fingers'll make yo' comfortable an' sociable; three'll raise the devil in
yo,' an' four'll make yo' dancin' drunk, when yo'll have t' be held
down. Yo'll have jest two fingers, an' not a drap more."
"Jest another finger, 'Frony. Remember, yo've bin orful rough on
me, an' I need more. I'll promise t' be good," pleaded Hackberry.
"No, not a drap more'n two fingers now. If yo' behave yo'self I'll
give yo' another two fingers by-an'-by."
"Hackberry swallowed his portion at a thirsty gulp and sat down on
the door-sill to let it do its invigorating work. The other two guerrillas
were given each two fingers, and the man whom Si knocked down
had his moanings rewarded by three fingers and a liberal application
in addition to the wound on his head, which he declared was much
relieved by it.
"Set your guns up agin the wall an' ack nacherul," commanded
Mrs. Bolster. "Nobody's a-gwine to hurt yo'. The 'Squire'll be here
soon, we'll git spliced, an' have a good time all around."
The noisy barking of the dogs announced the approach of
someone.
"Lord, I hope that's 'Squire Corson," said Mrs. Bolster, running
eagerly to the door. "If hit's him, we kin go right ahead with the
weddin'."
"If that's the 'Squire," said Shorty, in a low whisper, without turning
his head, "we'll grab our guns and fight to the death. We may clean
out this gang."
Si's attention had been in the meanwhile attracted to some boxes
concealed under the beds, and his curiosity was aroused as to what
such unusual things in a cabin might contain.
"No; hit's Capt. Sol. Simmons," said she in a tone of
disappointment mixed with active displeasure. "Now, he'll be cavortin'
and tearin' around, and wantin' t' kill somebody. I wish he wuz whar
hit's a good deal hotter."
She came over to where the boys were sitting, and said in a low
tone:
"This man's allers makin' trouble, an' he's bad from his boots up.
Keep a stiff upper lip, both on yo', an' we'll try t' manage him. Don't
weaken. Hit'll do no good. He'll be wuss'n ever then."
Si and Shorty instinctively felt for the revolvers in their pockets.
The newcomer tied his horse to a sapling and strode into the
house. The guerrillas seemed rather more fearful than otherwise to
see him, but met him with manners that were ranged from respectful
by Jeff Hackberry to absolute servility by the others. He was a burly,
black-bearded man, wearing a fairly-good uniform of a rebel Captain.
His face showed that he was a bully, and a cruel one.
He acknowledged in an overbearing way the greetings of the
others, and called out imperiously:
"'Frony, gi' me a stiff dram o' yer best at wunst. My throat's drier'n
a lime-kiln. Bin ridin' all mornin'."
"Folks wantin' likker don't say must t' me, but will yo', an' please,"
she answered sulkily.
"'Must,' 'please,' yo' hag," he said savagely. "Talk that a-way to me.
I'll 'please' yo'. I've killed two Yankees this mornin', an' I'm not in the
humor to fool around with an old pennyroyal huzzy like yo'. Gi' me
some whisky at wunst, or I'll baste yo'."
If ever Mrs. Bolster had been favorably disposed to him, she could
not endure to have him treat her this way before Shorty. She would
assert herself before him if ever.
 She put her arms akimbo and retorted vigorously:
"Nary drap o' likker yo'll git from me, Sol. Simmons. Go and git yer
likker whar y're welcome. Y're not welcome here. I don't keer if yo'
have killed two Yankees or 20 Yankees. Y're allers talkin' about killin'
Yankees, but nobody never sees none that y've killed. I'm a better
Confederit than yo' ever dared be. I'm doin' more for the Southern
Confedrisy. Y're allers a-blowin' while I'm allers adoin.' Everybody
knows that. Talk about the two Yankees y've killed, an' which
nobody's seed, here I've brung two Yankees right outen their camps,
an' have 'em to show. More'n that, they're gwine to jine we'uns."
She indicated the two boys with a wave of her hand. Simmons
seemed to see them for the first time.
"Yankees here, an' yo' haint killed 'em," he yelled. He put his hand
to his revolver and stepped forward. The two boys jumped up and
snatched their guns, but before another move could be made Mrs.
Bolster's unfailing trip brought Simmons heavily to the floor, with his
revolver half out the holster. In an instant she sat down heavily upon
him, and laid her brawny hand upon his pistol. The dogs and children
gathered around in joyous expectation of a renewal of general
hostilities. But the dogs broke away at the scent or sight of someone
approaching.
"Mebbe that's 'Squire Corson,'" said Mrs. Bolster with a renewed
flush of pleasant anticipation.
Instead, a rather, good-looking young rebel officer wearing a
Major's silver stars dismounted from his horse and, followed by two
men, entered the cabin.
"Hello, Simmons," said the Major in a tone of strong rebuke as
soon as he entered. "What in the world are you doing here? Is this
the way you carry out the General's orders? You're at your old tricks
again. You were sent out here early this morning, to capture or drive
away that Yankee picket at Raccoon Ford, so as to let Capt. Gillen
come through with his pack-mules. I expected to meet him here and
go on with him. Your men have been waiting at the crossroads for
you since daylight, while you've been loitering around the rear. I
ought to have you shot, and you would be if I reported this to the
General. You skulking whelp, you ought to be shot. But I'll give you
one more chance. It may not be too late yet. Break for your place as
fast as you can, and take these whelps with you. I'll wait here till
sundown for you. If you don't report back to me by that time you'd
better make your will. Jump now."
Mrs. Bolster had let go of Simmons as this exordium proceeded, as
she felt that he was in good hands.
As they disappeared the Major turned to Mrs. Bolster and inquired:
"Did Capt. Gillen get through with that quinine and guncaps?"
"They're thar," she said, pointing to the boxes under the beds.
"Very good. I've brought some men to take them away. We need
them very badly. Who are these men?"
Mrs. Bolster told her story about how they were tired of the
Abolition war, and had yielded to her persuasions to join the
Southern army.
The Major looked them over sharply, and began a close cross-
questioning as to where they were born, what regiment they
belonged to, how long they had been in the service, what battles
they had been engaged in and on what part of the field, where their
regiment now was, its brigade, division and corps, commanders, etc.,
etc.
As Shorty did not see any present occasion for lying, he had no
trouble in telling a convincing straightforward story. Si successfully
worked the loss-of-voice racket, and left the burden of conversation
to his partner.
The Major seemed satisfied, and said at the conclusion:
"Very good. I'll take you back with me when I return, and place
you in a good regiment."
This was a new and startling prospect, which was almost too much
for Shorty's self-control. For a minute he had wild thoughts of
assassinating the Major then and there, and making a run for life.
But he decided to wait a little longer and see what would develop.
 If Mrs. Bolster's hue had permitted she would have turned pale at
this threatened loss of a husband and upsetting of all her plans. She
merely gulped down a lump in her throat and seemed to be thinking.
She became very attentive to the Major, and brought for his
edification a private bottle of fine old whisky. She set about preparing
something for them to eat.
Again the dogs barked, and in walked a man dressed in the fatigue
uniform of a Union soldier with the chevrons of a Sergeant. The boys
gave a start of surprise, and a great one when they saw on his cap:

A
200 Ind. Vols.

Si would have sprung up to greet him, but Shorty laid a restraining

hand, and whispered:
"He don't belong to our regiment."
A second glance satisfied Si of this. While it is hardly possibly for a
man to know every other man in his regiment, yet in a little while
there comes something which enables him to know whether any man
he meets does or does not belong to his regiment.
The Major and Mrs. Bolster instantly recognized the newcomer.
"Awful glad to see you, Tuggers," said the Major, rising and shaking
his hand. "Did you get through without any trouble?"
"Not a bit o' trouble, thanks to you and Mrs. Bolster here. She got
me this uniform and this cap," said Tuggers, taking off the latter
article and scanning the lettering. "Rather more brass than I'm in the
habit of carrying on top of my head, no matter how much I have in
my face. I got your not giving me the positions of the Yankee
regiments, for which I suppose we must also thank Mrs. Bolster. I
found them all correct. As the 200th Ind. was the farthest out, I had
no difficulty getting through the rest of them by saying that I was on
my way to my regiment. Of course, I didn't come through the camp
of the 200th Ind., but modestly sought a byroad which Mrs. Bolster
had put me onto. I've got a lot of important letters from the mail in
Nashville, among which are some letters for the General, which I am
told are highly important. I'm mighty glad to be able to place them in
your hands, and relieve myself of the responsibility. Here they are.
Thanks, I don't care if I do, since you press me so hard," said he,
without change of voice, as he handed over the letters and picked up
the bottle and tin cup.
"Excuse me, Tuggers, for not asking you before," said the Major. "I
was so interested in you and your letters I forgot for the moment
that you might be thirsty. Help yourself."
"I didn't forget it," said Tuggers, pouring out a liberal dram. "Here's
to our deserving selves and our glorious Cause."
A shy girl of about eight had responded to Si's persistent
encouragement, and sidled up to him, examining his buttons and
accouterments. Si gave her some buttons he had in his pocket, and
showed her his knife and other trinkets in his pockets. The other
children began to gather around, much interested in the elaborate
dumb show he was making of his inability to speak.
Again the dogs barked. Mrs. Bolster ran to the door. "Hit's 'Squire
Corson," she exclaimed joyously, and hustled around to make extra
preparations for his entertainment.
The 'Squire entered, mopping his face with his bandana, and
moving with the deliberation and dignity consistent with his official
position.
He looked at the boys with a severe, judicial eye, and gave the
ominous little cough with which he was wont to precede sentences.
But he recognized the Major and Tuggers, and immediately his
attention was centered in them. They were connected with Army
Headquarters; they were repositories of news which he could spread
among his constituents. He greeted them effusively, and was only too
glad to accept their invitation to sit down and drink. But he
suggested, with official prudence, that they go out in front and sit
under a tree where they could converse wore at liberty.
"Afore you go out, 'Squire," said Mrs. Bolster, with an attempt at
coyness, "I want yo' t' do a little job fer me."
Shorty's hair tried to stand on end.
 "Jest wait a little, my good woman," said the 'Squire patronizingly.
"I want to talk to these gentlemen first; I kin 'tend to your matter
any time."
They lighted their pipes, and talked and talked, while Mrs. Bolster
fidgeted around in growing anxiety. Finally, as the sun was going
down, she could stand it no longer, and approached the group.
"'Squire," she said, "I'm orferly anxious to have a little job o' mine
done. 'Twon't take yo' five minits. Please 'tend to it right away."
"What is it she wants?" inquired the Major.
"I think she wants me to marry her to a Yankee deserter in there.
She whispered suthin' o' that kind to me awhile ago."
"That reminds me," said the Major; "I want you to swear those two
men into the service of the Southern Confederacy. You might as well
do it now, if you please, for I want to take them back with me and
put them into a regiment."
"That won't give much of a honeymoon to Mrs. Bolster," grinned
the 'Squire.
"Well, we've all got to make sacrifices for the Cause," said the
Major; "her honeymoon'll be the sweeter for being postponed. I've
had to postpone mine."
"Well, bring the men out," said the 'Squire, pouring himself out
another drink.
Si and Shorty had moved to the front door when Mrs. Bolster went
out, and could hear the whole conversation. They looked at one
another. Their faces were whiter than they had ever been on the field
of battle.
"Take the oath of allegiance to the Southern Confederacy? Die
right here a hundred times," surged through both their hearts.
Si pulled the bunches of firecrackers from his pocket, undid them
before the children's wondering eyes. He went through a pantomime
to tell them to take a coal from the fire, run out back with them, and
touch it to the fuses.
 "Take a coal, run back, and tech it to them strings," said Shorty,
forgetting himself in his excitement. "It'll be the greatest fun ye ever
saw."
"What's that y're sayin'?" said Mrs. Bolster.
"Jest talkin' to the children," said Shorty, seeing with relief the
children bolt out of the back door. He slipped his hand on his
revolver, determined to kill the 'Squire, the Major, and the other three
men before he would take a syllable of the oath.
"Come out here, men," said the Major authoritatively. Si slipped his
hand into his pocket, grasped his revolver, and walked forward very
slowly.
"Ahem," said the 'Squire, with an official cough. "Raise yer right
hands, and repeat these words after me, givin' your own names."
The other rebels took off their hats.
The dogs raised a clamor, which directed all eyes to the road. Sol
Simmons and the rest could be seen coming on a dead run.
"What does that mean?" said the Major anxiously.
At the same instant there was a series of crashes behind the
house; the firecrackers were going off like a volley of rifle-shots. The
Major whirled around to see what that meant, and looked into the
muzzle of Shorty's revolver.
"Surrender, or I'll kill you," shouted Shorty desperately. "Don't stop
a minit. Throw up your hands, I tell you."
Si was making a similar demand on Tuggers, while the 'Squire was
standing, open-mouthed, with the first word of the oath apparently
still on his tongue.
The Major sprang at Shorty, whose bullet cut his hair. The next
bullet caught the officer in the shoulder, and he reeled and went
down. Si was not so fortunate with Tuggers, who succeeded in
grappling him. Simmons dashed by and struck Si, in passing, with his
fist, which sent him to the ground, with Tuggers on top.
The next minute the 'Squire, who was the only one who had any
opportunity to look, saw Yankees pop out of the brush and jump the
fences in a long, irregular line which immediately surrounded the
house. Capt. McGillicuddy cut down Simmons with his sword, and the
rest incontinently surrendered.
"We had got tired of waiting, and were on the point of dashing in,
anyhow, when we heard the firecrackers," said Capt. McGillicuddy,
after the prisoners had been secured and things quieted down. "That
feller that I cut down was out there with a squad and caught sight of
us, and started back this way, and I concluded to follow him up and
jump the house. Neither of you hurt, are you?"
"Not hurt a mite," answered Shorty cheerfully, "but it's the closest
squeak I ever had. Wouldn't go through it agin for a pile o'
greenbacks big as a cornshock. Say, Cap., you've made a ten-strike
today that ought to make you a Major. That house's plum full o'
contraband, and there's a lot o' important letters there. But, say,
Cap., I want you to either kill that 'Squire or git him as fur away as
possible. I ain't safe a minnit as long as him and that woman's a-nigh
me."
 CHAPTER XVI. AN UNEXPECTED
MARRIAGE
THE BOYS CAPTURE REBELS AND ADMINISTER
THE OATH.
THE REBEL Major accepted the unexpected turn of events with
soldierly philosophy. Tuggers, captured in a blue uniform, saw the
ignominious fate of a spy loom up before his eyes. His face grew very
white and set. He sat down on a log, looked far away, and seemed
oblivious to everything around him.
Jeff Hackberry and Sol Simmons were frightened into nerveless
terror, and occasionally sighed and groaned audibly. Their men
huddled together like frightened sheep, and looked anxiously at
every move of their captors.
'Squire Corson had ventured two or three remarks in a judicial and
advisory way, but had been ordered by Capt. McGillicuddy to sit
down and keep quiet. He took a seat on a stump, pulled a large
bandana out of his beaver crowned hat, wiped his bald head, and
anxiously surveyed the scene as if looking for an opportunity when
the power and dignity of the State of Tennessee might be invoked to
advantage.
Only Mrs. Bolster retained her aggressiveness and her tongue. If
anything, she seemed to be more savage and virulent than ever. She
was wild that she had been outwitted, and particularly by Si, whose
fluent speech had returned the moment the firecrackers went oif. She
poured out volleys of scorching epithets on all the Yankees from
President Lincoln down to Corp'l Si Klegg, and fervently invoked for
them speedy death and eternal torment where the worm dieth not
and the fire is not quenched.
 Capt. McGillicuddy rounded up his prisoners, took arms from those
who still retained them, had Si and Shorty do what they could toward
dressing the Major's wound, and then began an examination of the
house.
He found abundant evidence of all that he, Si and Shorty had
believed of it. It was a rendezvous for spies, both great and small—
both those, like Mrs. Bolster, who infested our camps, and got news
of whatever was going on there, and those who operated on a larger
scale, passing directly from the Headquarters of the rebels to the
Headquarters of ours, and to the rear, and the sources of information
at Nashville and Louisville. It was an important station on the route
for smuggling gun-caps, quinine, medicines and other contraband
from the North. Quantities of these were there waiting to be
forwarded. As the source of the fighting whisky introduced into the
camp of the 200th Ind. too much was known of it to require any
further information. And it was more than probable that it was the
scene of darker crimes—Union soldiers lured thither under some
pretext, murdered and robbed.
"How in the world am I going to break this infernal nest up?" said
Capt. McGillicuddy, with a puzzled air, after he had ordered the
whisky destroyed and the other things gotten in shape to send back
to camp. "By rights, I ought to burn that house down, but that would
leave all these children without shelter. By the same token, I ought to
shoot or at least send off to prison that old she-catamount, but that
would mean starving the children to death. I declare, I don't know
what to do."
He had drawn apart a little with Si and Shorty, to whom he spoke
confidentially, while casting his eyes about him as if seeking some
solution of the problem.
"If you'll allow me. Captain," said Shorty, "I've an idee. Now that
we've got the trap, let's set it agin, and see if we can't ketch some
more."
"Splendid idea. Shorty," said the Captain, catching on at once.
"And my idee," said Shorty, emboldened by the reception of his
first suggestion, "is that you take all the company but me and Si and
four or fire of the boys back to camp, leavin' us here until to-morrow
at least. There'll probably some very interestin' men happen along
here to-night, not knowing what's happened, and we'll jest quietly
yank 'em in."
"That's good," assented the Captain.
"In the meantime," continued Shorty, "you kin be considerin' what
you'll do with the house. It may be best to let it stand, and watch it.
That's a good way to do with a bee-tree or a woodchuck hole.
"I believe you are right. I'll do as you say. Si, you and Shorty pick
out as many men as you want to stay with you. I'll leave one of
these horses with you. If you should happen to need any more,
mount one of the boys and send him back for help. I'll come out with
the whole company."
Shorty and Si consulted together for a few minutes, picked out
their men, gave their names to the Captain, and received his assent
to the selection. Then Shorty said:
"Captain, you don't want to take that old woman, the 'Squire and
that skunk they call Jeff Hackberry back to camp with you, do you?
Leave 'em here with us. I've got a little scheme."
"The old woman and the 'Squire you can take and welcome,"
answered the Captain. "I'll be glad to have them oif my hands. But
Hackberry is a rebel soldier. I don't know about giving him up."
"Leave him with us, then. We'll turn him back to you all right, and
the old woman and the 'Squire, too, if you want 'em."
"No," said the Captain, with an impatient wave of his hand. "Keep
them, do what you please with them. If you should accidentally kill
the old woman I should not be unduly distressed. But don't let
Hackberry get away from you. I'll take the rest back to camp, and I
must start at once, for it's getting late, and we didn't bring any
rations with us. Do you suppose you can find enough around the
house to keep you till morning?"
"O, yes," said Si. "There's a sack of meal in there and some side-
meat. We gave the old woman a lot of coffee. We'll make out all
right."
 The prisoners had been watching the Captain and his men with
greatest anxiety. They now saw Si with his squad take the 'Squire.
Mrs. Bolster and Hackberry off to one side, while the Captain placed
the remainder of the prisoners in the center of his company and
started back to camp with them.
There was something in this separation that terrified even Mrs.
Bolster, who stopped railing and began to look frightened.
"What are yo'uns goin' to do with we'uns?" she inquired hoarsely
of Si.
"You'll find out soon enough," said Si significantly. "Set down there
on that log and think about what you deserve. You might put in any
spare time you have in doing some big repentin'."
Hackberry began to whine and beg for mercy, but Shorty ordered
him to keep silent.
"I want you to understand," said the 'Squire, "that I'm a regerlarly
elected and qualified Magistrate o' the State o' Tennessee; that I'm
not subjeck to military laws, and if any harm comes to me you'll have
to answer for it to the State o' Tennessee."
"Blast the State o' Tennessee," said Shorty contemptuously. "When
we git through there won't be no State o' Tennessee. It'll be roasting
in the same logheap with South Caroliny and Virginny, with Jeff Davis
brilin' in the middle."
"Boys," ordered Si, "a couple of you look around the house and see
if you can't find a mattock and shovel."
Terrible fears assailed the three unhappy prisoners at this. What
could a mattock and shovel be wanted for but to dig their graves?
Shorty stepped over a little distance to a large clump of "red-
sticks." These grow in long wands of brilliant red, as straight as a
corn-stalk, and slenderer. They are much used about the farms of the
South for rods for rough measurement. He cut one off about six feet
long and stripped off its leaves.
The anxious eyes of the prisoners followed every movement.
Two of the boys appeared with an old mattock and shovel.
 "Guess you'd better dig right over there," said Si, indicating a little
bare knoll.
"Nothin' else's ever bin planted there. At least nothin's ever come
up. The chances are agin their comin' up if we plant 'em there."
"Stand up," said Shorty, approaching Hackberry with the bright
crimson rod in his hand. "I'm goin' to measure you for a grass-green
suit that'll last you till Gabriel blows his horn."
Hackberry gave a howl of terror. The 'Squire and Mrs. Bolster
began a clamor of protests.
"Don't fuss," said Shorty calmly to them, as he took Hackberry's
dimensions. "I ain't goin' to show no partiality. I'll serve you both the
same way. Your turns 'll come after his'n."
The children, aware that something unusual was going on, yet
unable to comprehend what it was, stood silently around, their
fingers in their mouths and their vacant eyes fixed in the stolid stare
of the mountaineer youth. Even the dogs were quiet, and seemed
watching the scene with more understanding than the children.
Mrs. Bolster's mood suddenly changed from bitter vituperation. She
actually burst into tears, and began pleading for her life, and making
earnest promises as to better conduct in the future. The 'Squire and
Hackberry followed suit, and blubbered like schoolboys. Mrs. Bolster
reminded Si and Shorty how she had saved them from being killed by
the fierce Hackberry and the still fiercer Simmons. This seemed to
move them. She tried a ghastly travesty of feminine blandishments
by telling Shorty how handsome she had thought him, and had fallen
in love with him at first sight. Shorty gave a grimace at this. He and
Si stepped back a little for consultation.
When they came back Shorty said oracularly:
"Our orders is strict, and we should've carried 'em out at once.
But, talkin' with my partner here, we're reminded o' somethin'. We
believe it's the law that when a man or woman is sentenced to death
the execution kin be put off if they kin find anybody to marry 'em. Is
that good law, 'Squire?"
 "H-m-m," answered the Magistrate, resuming his judicial manner
at once; "that is a general belief, and I've heard o' some instances of
it. But before sayin' positively, I should like to examine the authorities
an' hear argument."
"Well, there hain't goin' to be no continuance in this case for you to
look up authorities and hear arguments," said Shorty decisively.
"We're the higher court in this case, and we decided that the law's
good enough for it. We've settled that if Mrs. Bolster 'll marry
Hackberry, and Hackberry 'll marry Mrs. Bolster, and you'll marry 'em
both, we'll grant a stay o' proceedings in the matter o' the execution
o' the sentence o' death until we kin be advised by the higher
authorities."
"I'll do anything. Mister," blubbered Hackberry. "I'll marry her this
minnit. Say the words, 'Squire."
"I've said I'd rather die 10 times over than marry yo', Jeff
Hackberry," murmured Mrs. Bolster. "I've bin the wife o' one ornery
snipe of a whisky-sucking sand-digger, and when the Lord freed me
from him I said I'd never git yoked with another. But I s'pose I've got
to live for my children, though the Lord knows the yaller-headed
brats hain't wuth hit. They're everyone of 'em their dad over agin—all
Bolsters, and not wuth the powder to blow 'em to kingdom come. I'd
a heap ruther marry Jeff Hackberry to make sure o' havin' him shot
than to save him from shootin'."
"You hain't no choice, Madam," said Shorty severely. "Law and
orders is strict on that pint."
"Well, then," said she, "since hit's a ch'ice betwixt death and Jeff
Hackberry, I'll take Jeff Hackberry, though I wouldn't take him on no
other terms, and I'm afeared I'm makin' a mistake as hit is."
"What do you say, 'Squire?" asked Shorty.
"I've bin studyin' on jest whar I come in," answered the
Magistrate. "These two save their necks by marryin', but do you
understand that the law says that the Magistrate who marries 'em
gits his neck saved?"
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