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TH I R D ED ITI O N

Microbiology
An Evolving Science

Joan L. Slonczewski
Kenyon College

John W. Foster
University of South Alabama

Appendices 1 and 2 by
Kathy M. Gillen
Kenyon College

W. W. NORTON & COMPANY

NEW YORK LONDON
W. W. Norton & Company has been independent since its founding in 1923, when William Warder
Norton and Mary D. Herter Norton first published lectures delivered at the People’s Institute, the
adult education division of New York City’s Cooper Union. The firm soon expanded its program
beyond the Institute, publishing books by celebrated academics from America and abroad. By mid-
century, the two major pillars of Norton’s publishing program—trade books and college texts—
were firmly established. In the 1950s, the Norton family transferred control of the company to its
employees, and today—with a staff of four hundred and a comparable number of trade, college, and
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Copyright © 2014, 2011, 2009 by W. W. Norton & Company, Inc.

All rights reserved.
Printed in the United States of America.
Third Edition

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Manufacturing by Transcontinental–Beauceville QC

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 Dedication
We dedicate this Third Edition to the memory of Lynn Margulis
(1938–2011) and Carl Woese (1928–2012), who forever
changed our understanding of evolutionary biology. Margulis
hypothesized that modern-day eukaryotic cells evolved from
symbiotic relationships with bacteria; and Woese used the
evidence of gene sequences to propose the now accepted
paradigm of three, continually evolving, domains of life: Bacteria,
Archaea, and Eukarya. The world will long miss the deep intellect
and larger-than-life personalities of these two scientists. We, the
authors, were profoundly influenced by their thinking and deeply
moved by their passing.
BRIEF CONTENTS

eTopic Contents!xix PART 4

Preface!xxi
Microbial Diversity
About the Authors!xxxiii
and Ecology'662
PART 1 17 Origins and Evolution!665
18 Bacterial Diversity!711
The Microbial Cell'2 19 Archaeal Diversity!757
1 Microbial Life: Origin and Discovery!5
20 Eukaryotic Diversity!797
2 Observing the Microbial Cell!41
21 Microbial Ecology!839
3 Cell Structure and Function!79
22 Microbes in Global Elemental Cycles!889
4 Bacterial Culture, Growth, and
Development!119
5 Environmental Influences and Control PART 5
of Microbial Growth!157 Medicine and Immunology'922
6 Viruses!191
23 Human Microbiota and Innate Immunity!925
24 The Adaptive Immune Response!961
PART 2 25 Microbial Pathogenesis!1003
Genes and Genomes'234 26 Microbial Diseases!1051

7 Genomes and Chromosomes!237 27 Antimicrobial Therapy!1107

8 Transcription, Translation, 28 Clinical Microbiology and Epidemiology!1147

and Bioinformatics!275
9 Gene Transfer, Mutations, and Genome APPENDIX 1: Biological Molecules!A-1
Evolution!321 APPENDIX 2: Introductory Cell Biology:
10 Molecular Regulation!365 Eukaryotic Cells!A-23
11 Viral Molecular Biology!409 APPENDIX 3: Laboratory Methods for
Microbiology!A-43
12 Biotechniques and Synthetic Biology!457
APPENDIX 4: Taxonomy!A-59

PART 3 Answers to Thought Questions!AQ-1

Metabolism and Glossary!G-1

Biochemistry'488 Figure Credits!F-1

Index!I-1
13 Energetics and Catabolism!491
14 Electron Flow in Organotrophy, Lithotrophy,
and Phototrophy!539
15 Biosynthesis!585
16 Food and Industrial Microbiology!623

VI
 CONTENTS

eTopic Contents!xix
Preface!xxi
About the Authors!xxxiii

PART 1
The Microbial Cell 2
AN INTERVIEW WITH RICHARD LENSKI:
Evolution in the Lab

CHAPTER 1
Microbial Life: Origin and Discovery........................................................................ 5
1.1 From Germ to Genome: What Is a Microbe?!7
1.2 Microbes Shape Human History!10
Special Topic 1.1: How Did Life Originate?!18
1.3 Medical Microbiology!20
1.4 Microbial Ecology!26
1.5 The Microbial Family Tree!29
1.6 Cell Biology and the DNA Revolution!32

CHAPTER 2
Observing the Microbial Cell ....................................................................................41
2.1 Observing Microbes!42
2.2 Optics and Properties of Light!46
2.3 Bright-Field Microscopy!51
2.4 Fluorescence Microscopy!58
2.5 Dark-Field and Phase-Contrast Microscopy!63
2.6 Electron Microscopy and Tomography!66
2.7 Visualizing Molecules!73
Special Topic 2.1: Molecular “Snapshots”: Chemical Imaging!74

VII
VIII ! CONTENTS

CHAPTER 3
Cell Structure and Function .....................................................................................79
3.1 The Bacterial Cell: An Overview!81
3.2 The Cell Membrane and Transport!86
3.3 The Cell Wall and Outer Layers!91
3.4 The Nucleoid, RNA, and Protein Synthesis!101
3.5 Cell Division!104
3.6 Cell Polarity and Aging!107
Special Topic 3.1: Senior Cells Make Drug-Resistant Tuberculosis!108
3.7 Specialized Structures!112

CHAPTER 4
Bacterial Culture, Growth, and Development ................................................. 119
4.1 Microbial Nutrition!120
4.2 Nutrient Uptake!127
4.3 Culturing Bacteria!133
4.4 Counting Bacteria!136
4.5 The Growth Cycle!139
4.6 Biofilms!145
Special Topic 4.1: Sharks and Biofilms Don’t Mix!146
4.7 Cell Differentiation!150

CHAPTER 5
Environmental Influences and Control of Microbial Growth ..................... 157
5.1 Environmental Limits on Growth!158
5.2 Adaptation to Temperature!160
Special Topic 5.1: It’s Raining Bacteria!163
5.3 Adaptation to Pressure!164
5.4 Water Activity and Salt!166
5.5 Adaptation to pH!167
5.6 Oxygen and Other Electron Acceptors!172
5.7 Nutrient Deprivation and Starvation!176
5.8 Physical, Chemical, and Biological Control of Microbes!178

CHAPTER 6
Viruses ........................................................................................................................... 191
6.1 What Is a Virus?!192
6.2 Virus Structure!198
6.3 Viral Genomes and Classification!203
6.4 Bacteriophage Replication!208
6.5 Animal and Plant Virus Replication!213
 CONTENTS ! IX

6.6 Culturing Viruses!221

Special Topic 6.1: The Good Viruses!226
6.7 Viral Ecology!226

PART 2
Genes and Genomes 234

AN INTERVIEW WITH CHRISTINE JACOBS-WAGNER:

The Thrill of Discovery in Molecular Microbiology

CHAPTER 7
Genomes and Chromosomes ................................................................................ 237
7.1 DNA: The Genetic Material!238
7.2 Genome Organization!240
7.3 DNA Replication!248
7.4 Plasmids!258
7.5 Eukaryotic Chromosomes!260
7.6 DNA Sequence Analysis!263
Special Topic 7.1: Where Have All the Bees Gone? Metagenomics, Pyrosequencing, and Nature!268

CHAPTER 8
Transcription, Translation, and Bioinformatics .............................................. 275
8.1 RNA Polymerases and Sigma Factors!276
8.2 Transcription of DNA to RNA!280
8.3 Translation of RNA to Protein!286
Special Topic 8.1: Stalking the Lone Ribosome!298
8.4 Protein Modification and Folding!303
8.5 Protein Degradation: Cleaning House!304
8.6 Secretion: Protein Traffic Control!306
8.7 Bioinformatics: Mining the Genomes!312

CHAPTER 9
Gene Transfer, Mutations, and Genome Evolution ....................................... 321
9.1 The Mosaic Nature of Genomes!322
9.2 Gene Transfer!322
Special Topic 9.1: There’s a Bacterial Genome Hidden in My Fruit Fly!330
X ! CONTENTS

9.3 Recombination!338
9.4 Mutations!341
9.5 DNA Repair!348
9.6 Mobile Genetic Elements!355
9.7 Genome Evolution!358

CHAPTER 10
Molecular Regulation ............................................................................................... 365
10.1 Regulating Gene Expression!366
10.2 Paradigm of the Lactose Operon!369
10.3 Other Systems of Operon Control!376
10.4 Sigma Factor Regulation!382
10.5 Regulatory RNAs!385
10.6 DNA Rearrangements: Phase Variation by Shifty Pathogens!389
10.7 Integrated Control Circuits!391
10.8 Quorum Sensing: Chemical Conversations!397
10.9 Transcriptomics and Proteomics!401
Special Topic 10.1: Networking with Nanotubes!402

CHAPTER 11
Viral Molecular Biology ...........................................................................................409
11.1 Phage T4: The Classic Molecular Model!410
11.2 Hepatitis C: (+) Strand RNA Virus!417
11.3 Influenza Virus: (–) Strand RNA Virus!424
11.4 Human Immunodeficiency Virus (HIV): Retrovirus!432
11.5 Herpes Simplex Virus: DNA Virus!443
Special Topic 11.1: Cytomegalovirus!448
11.6 Gene Therapy with Viruses!450

CHAPTER 12
Biotechniques and Synthetic Biology ................................................................ 457
12.1 Basic Tools of Biotech: A Research Case Study!458
12.2 Genetic Analyses!458
12.3 Classic Molecular Techniques!463
12.4 Viewing the Interactions and Movements of Proteins!471
12.5 Applied Biotechnology!474
12.6 Synthetic Biology: Biology by Design!477
Special Topic 12.1: Bacteria “Learn” to Keep Time and Signal Danger!478
 CONTENTS ! XI

PART 3
Metabolism and Biochemistry 488

AN INTERVIEW WITH DAN WOZNIAK:

Polymer Biosynthesis Makes a Pathogenic Biofilm

CHAPTER 13
Energetics and Catabolism .................................................................................... 491
13.1 Energy and Entropy for Life!494
13.2 Energy in Biochemical Reactions!498
13.3 Energy Carriers and Electron Transfer!501
Special Topic 13.1: Microbial Syntrophy Cleans Up Oil!502
13.4 Catabolism: The Microbial Buffet!510
13.5 Glucose Breakdown and Fermentation!516
13.6 The Tricarboxylic Acid (TCA) Cycle!527
13.7 Aromatic Pollutants!532

CHAPTER 14
Electron Flow in Organotrophy, Lithotrophy, and Phototrophy .............. 539
14.1 Electron Transport Systems!541
14.2 The Proton Motive Force!546
Special Topic 14.1: Testing the Chemiosmotic Theory!548
14.3 The Respiratory ETS and ATP Synthase!551
14.4 Anaerobic Respiration in Organotrophs!559
Special Topic 14.2: Bacterial Electric Power!562
14.5 Lithotrophy and Methanogenesis!563
14.6 Phototrophy!571

CHAPTER 15
Biosynthesis ................................................................................................................585
15.1 Overview of Biosynthesis!586
15.2 CO2 Fixation: The Calvin Cycle!589
15.3 CO2 Fixation: Diverse Pathways!597
15.4 Biosynthesis of Fatty Acids and Polyketides!601
15.5 Nitrogen Fixation!605
Special Topic 15.1: Mining a Bacterial Genome for Peptide Antibiotics!606
15.6 Biosynthesis of Amino Acids and Nitrogenous Bases!612
15.7 Biosynthesis of Tetrapyrroles!618
XII ! CONTENTS

CHAPTER 16
Food and Industrial Microbiology ........................................................................623
16.1 Microbes as Food!625
16.2 Fermented Foods: An Overview!627
16.3 Acid- and Alkali-Fermented Foods!629
16.4 Ethanolic Fermentation: Bread and Wine!637
16.5 Food Spoilage and Preservation!641
16.6 Industrial Microbiology!650
Special Topic 16.1: Companies Take On Tuberculosis!652
Special Topic 16.2: Microbial Enzymes Make Money!654

PART 4
Microbial Diversity and Ecology 662

AN INTERVIEW WITH NICOLE DUBILIER:

Marine Animals with Bacterial Symbionts

CHAPTER 17
Origins and Evolution............................................................................................... 665
17.1 Origins of Life!667
17.2 Early Metabolism!676
17.3 Microbial Phylogeny and Gene Transfer!681
Special Topic 17.1: Phylogeny of a Shower Curtain Biofilm!686
17.4 Adaptive Evolution!692
17.5 Microbial Species and Taxonomy!696
Special Topic 17.2: Jump-Starting Evolution of a Hyperthermophilic Enzyme!698
17.6 Symbiosis and the Origin of Mitochondria and Chloroplasts!703

CHAPTER 18
Bacterial Diversity ..................................................................................................... 711
18.1 Bacterial Diversity at a Glance!712
18.2 Cyanobacteria: Oxygenic Phototrophs!718
18.3 Firmicutes and Actinobacteria (Gram-Positive)!723
18.4 Proteobacteria (Gram-Negative)!734
Special Topic 18.1: Carbon Monoxide: Food for Bacteria?!738
18.5 Deep-Branching Gram-Negative Phyla!748
18.6 Spirochetes: Sheathed Spiral Cells with Internalized Flagella!750
18.7 Chlamydiae, Planctomycetes, and Verrucomicrobia: Irregular Cells!752
 CONTENTS ! XIII

CHAPTER 19
Archaeal Diversity ..................................................................................................... 757
19.1 Archaeal Traits!758
19.2 Crenarchaeota across the Temperature Range!766
19.3 Thaumarchaeota: Symbionts and Ammonia Oxidizers!773
19.4 Methanogens!775
Special Topic 19.1: Eating Ammonia: Thaumarchaeotes!776
19.5 Haloarchaea!784
Special Topic 19.2: Haloarchaea in the Classroom!786
19.6 Thermophilic and Acidophilic Euryarchaeota!790
19.7 Deeply Branching Divisions!793

CHAPTER 20
Eukaryotic Diversity ................................................................................................. 797
20.1 Phylogeny of Eukaryotes!798
20.2 Fungi!806
Special Topic 20.1: Yeast: A Single-Celled Human Brain?!810
20.3 Algae!818
20.4 Amebas and Slime Molds!824
20.5 Alveolates: Ciliates, Dinoflagellates, and Apicomplexans!827
20.6 Trypanosomes and Metamonads!833
Special Topic 20.2: The Trypanosome: A Shape-Shifting Killer!834

CHAPTER 21
Microbial Ecology ...................................................................................................... 839
21.1 Metagenomes—and Beyond!841
21.2 Functional Ecology!849
21.3 Symbiosis!852
21.4 Marine and Aquatic Microbes!857
Special Topic 21.1: Cleaning Up the Deepwater Oil Spill!858
21.5 Soil and Subsurface Microbes!869
21.6 Plant Microbial Communities!876
21.7 Animal Microbial Communities!882

CHAPTER 22
Microbes in Global Elemental Cycles .................................................................889
22.1 Biogeochemical Cycles!890
22.2 The Carbon Cycle and Bioremediation!894
22.3 The Hydrologic Cycle and Wastewater Treatment!896
Special Topic 22.1: Bioremediation of Weapons Waste!902
22.4 The Nitrogen Cycle!903
22.5 Sulfur, Phosphorus, and Metals!908
22.6 Astrobiology!916
XIV ! CONTENTS

PART 5
Medicine and Immunology 922

AN INTERVIEW WITH FERRIC FANG:

Molecular Microbiology Dissects a Pathogen

CHAPTER 23
Human Microbiota and Innate Immunity........................................................... 925
23.1 Human Microbiota: Location and Shifting Composition!926
23.2 Risks and Benefits of Microbiota!934
23.3 Overview of the Immune System!935
Special Topic 23.1: Are NETs a Cause of Lupus?!940
23.4 Barbarians at the Gate: Innate Host Defenses!943
23.5 The Acute Inflammatory Response!946
23.6 How Phagocytes Detect and Kill Microbes!950
23.7 Interferon, Natural Killer Cells, and Toll-like Receptors!953
23.8 Complement’s Role in Innate Immunity!955
23.9 Fever!958

CHAPTER 24
The Adaptive Immune Response ......................................................................... 961
24.1 Overview of Adaptive Immunity!962
24.2 Immunogenicity!965
24.3 Antibody Structure and Diversity!968
24.4 Primary and Secondary Antibody Responses!974
24.5 Genetics of Antibody Production!977
24.6 T Cells Link Antibody and Cellular Immune Systems!982
24.7 Complement as Part of Adaptive Immunity!992
Special Topic 24.1: An Uneasy Peace: Détente at the Microbiota-Intestine Interface!994
24.8 Hypersensitivity and Autoimmunity!995

CHAPTER 25
Microbial Pathogenesis ........................................................................................ 1003
25.1 Host-Pathogen Interactions!1004
25.2 Virulence Factors and Pathogenicity Islands!1010
25.3 Microbial Attachment: First Contact!1013
25.4 Toxins Subvert Host Function!1018
 CONTENTS ! XV

25.5 Deploying Toxins and Effectors!1028

25.6 Surviving within the Host!1033
Special Topic 25.1: Type VI Secretion: Poison Darts!1034
25.7 Experimental Tools That Define Pathogenesis!1045

CHAPTER 26
Microbial Diseases ................................................................................................. 1051
26.1 Characterizing and Diagnosing Microbial Diseases!1052
26.2 Skin and Soft-Tissue Infections!1054
26.3 Respiratory Tract Infections!1058
26.4 Gastrointestinal Tract Infections!1066
Special Topic 26.1: Sprouts and an Emerging Escherichia coli!1068
26.5 Genitourinary Tract Infections!1075
26.6 Central Nervous System Infections!1083
26.7 Cardiovascular System Infections!1090
26.8 Systemic Infections!1094
26.9 Immunization!1102

CHAPTER 27
Antimicrobial Therapy .......................................................................................... 1107
27.1 The Golden Age of Antibiotic Discovery!1108
27.2 Fundamentals of Antimicrobial Therapy!1111
27.3 Measuring Drug Susceptibility!1112
27.4 Mechanisms of Action!1116
27.5 Challenges of Drug Resistance!1126
27.6 The Future of Drug Discovery!1134
27.7 Antiviral Agents!1135
Special Topic 27.1: Anti-Quorum Sensing Drug Blocks Pathogen “Control and Command”!1136
Special Topic 27.2: Resurrecting the 1918 Pandemic Flu Virus!1139
27.8 Antifungal Agents!1142

CHAPTER 28
Clinical Microbiology and Epidemiology .........................................................1147
28.1 Principles of Clinical Microbiology!1148
28.2 Specimen Collection and Processing!1149
28.3 Conventional Approaches to Pathogen Identification!1152
28.4 Rapid Techniques for Pathogen Identification!1160
28.5 Point-of-Care Rapid Diagnostics!1167
28.6 Biosafety Containment Procedures!1169
28.7 Principles of Epidemiology!1171
Special Topic 28.1: What’s Blowing in the Wind?!1178
28.8 Detecting Emerging Microbial Diseases!1179
XVI ! CONTENTS

APPENDIX 1
Biological Molecules .................................................................................................. A-1
A1.1 Elements, Bonding, and Water!A-2
A1.2 Organic Molecules!A-6
A1.3 Proteins!A-8
A1.4 Carbohydrates!A-11
A1.5 Nucleic Acids!A-13
A1.6 Lipids!A-15
A1.7 Biological Chemistry!A-16
Special Topic A1.1: Calculating the Standard Free Energy Change, ∆G°, of Chemical Reactions!A-19

APPENDIX 2
Introductory Cell Biology: Eukaryotic Cells ................................................... A-23
A2.1 The Cell Membrane!A-24
A2.2 The Nucleus and Mitosis!A-31
A2.3 Problems Faced by Large Cells!A-34
A2.4 The Endomembrane System!A-34
A2.5 The Cytoskeleton!A-38
A2.6 Mitochondria and Chloroplasts!A-40

APPENDIX 3
Laboratory Methods for Microbiology ............................................................A-43
A3.1 Isolating Parts of Cells by Using an Ultracentrifuge!A-44
A3.2 Agarose Gel Electrophoresis!A-45
A3.3 Protein Identification on 2D Gels with Mass Spectrometry!A-47
A3.4 RNA and DNA Identification by Northern and Southern Blots!A-49
A3.5 Sanger Method of DNA Sequencing!A-51
A3.6 Gene Fusions Identify Regulatory Mutants!A-52
A3.7 Primer Extension Identifies Transcriptional Start Sites!A-52
A3.8 DNA Microarray!A-54
A3.9 Multiplex PCR!A-55
A3.10 Fluorescence In Situ Hybridization (FISH) and CARD-FISH!A-55
A3.11 Immunoprecipitation Techniques!A-57

APPENDIX 4
Taxonomy ................................................................................................................... A-59
A4.1 Viruses!A-60
A4.2 Bacteria!A-62
A4.3 Archaea!A-66
A4.4 Eukarya!A-68

Answers to Thought Questions!AQ-1

Glossary!G-1
Figure Credits!F-1
Index!I-1
 eTOPIC CONTENTS
Access to the eTopics is available through both the ebook and the Norton Coursepack.

1.1 An Interview with Rita Colwell: The Global Impact 10.5 The Phage Lambda Lysis/Lysogeny “Decision”
of Microbiology Is to Kill or Not to Kill
1.2 Discovering the Genetic Code 11.1 Poliovirus: (+) Strand RNA
1.3 Clifford W. Houston: From Aquatic Pathogens 11.2 The Filamentous Phage M13: Vaccines
to Outer Space—An Interview and Nanowires
2.1 Confocal Microscopy 11.3 Genetic Resistance to HIV
3.1 Isolation and Analysis of the Ribosome 12.1 Mapping the E. coli Interactome
3.2 How Antibiotics Cross the Outer Membrane 12.2 GFP Proteins Track Cell Movements in Biofilms
3.3 Outer Membrane Proteins: Isolation for Vaccine 12.3 DNA Vaccines
3.4 Experiments That Reveal the Bacterial Cytoskeleton 12.4 Gene Therapy and Gene Delivery Systems
4.1 Transport by Group Translocation: 12.5 Directed Evolution through Phage Display
The Phosphotransferase System Technology
4.2 Eukaryotes Transport Nutrients by Endocytosis 12.6 DNA Shuffling Enables In Vitro Evolution
4.3 Biofilms, Antibiotics, Garlic, and Disease 12.7 Site-Directed Mutagenesis Helps Us Probe Protein
5.1 The Arrhenius Equation Function
5.2 Some Alkaliphilic Enzymes Produce Useful Drug 13.1 Observing Energy Carriers in Living Cells
Delivery Systems 13.2 Swiss Cheese: A Product of Bacterial Catabolism
5.3 Signaling Virulence 13.3 Genomic Analysis of Metabolism
5.4 Oligotrophs 13.4 Pyruvate Dehydrogenase Connects Sugar
6.1 How Did Viruses Originate? Catabolism to the TCA Cycle
6.2 West Nile Virus, an Emerging Pathogen 13.5 Genetic Analysis of Aromatic Catabolism
7.1 Genes and Proteins Involved in DNA Replication 14.1 Caroline Harwood: A Career in Bacterial
Photosynthesis and Biodegradation: An Interview
7.2 Trapping a Sliding Clamp
14.2 Measuring Dy and DpH in Microbes by the Uptake
7.3 Replication Mechanisms of Bacteriophages
of Molecules
7.4 Plasmid Partitioning and Addiction
14.3 Environmental Regulation of the ETS
7.5 Equilibrium Density Gradient Centrifugation
14.4 ATP Synthesis at High pH
8.1 Building the Ribosome Machine
15.1 The Discovery of 14C
8.2 Discovering the mRNA Ribosome-Binding Site
15.2 Metagenomic Screening for Polyketide Drugs
8.3 Ubiquitination: A Ticket to the Proteasome
15.3 Antibiotic Factories: Modular Biosynthesis
8.4 What Is the Minimal Genome? of Vancomycin
9.1 F Pili and Biofilm Formation 15.4 Riboswitch Regulation
9.2 Mapping Bacterial Chromosome Gene Position 16.1 From Barley and Hops to Beer
by Conjugation
16.2 Caterpillar Viruses Produce Commercial Products
9.3 Deinococcus Uses RecA to Repair Fragmented
17.1 The RNA World: Clues for Modern Medicine
Chromosomes
17.2 Horizontal Gene Transfer in E. coli O157:H7
9.4 Mutation Rate
17.3 Leaf-cutter Ants with Partner Fungi and Bacteria
9.5 The Transposase for a Bacterial Transposon
Resembles the Integrase for HIV-1 18.1 Karl Stetter: Adventures in Microbial Diversity Lead
to Products in Industry
9.6 Integrons and Gene Capture
20.1 Oomycetes: Lethal Parasites That Resemble Fungi
10.1 CRP Interactions with RNA Polymerase
and CRP-Dependent Promoters 20.2 A Ciliate Model for Human Aging
10.2 Glucose Transport Alters cAMP Levels 21.1 Mapping Bermuda Phytoplankton
10.3 Slipped-Strand Mispairing 21.2 Cold-Seep Ecosystems
10.4 Toxin-Antitoxin Modules: Mechanisms 22.1 Wetlands: Disappearing Microbial Ecosystems
for Self-preservation or Altruism? 22.2 Metal Contamination and Bioremediation

XVII
XVIII ! ETOPIC CONTENTS

23.1 Microbes as Vaccine Delivery Systems 26.1 Sequenced Genomes of Pathogens

23.2 Do Defensins Help Determine Species Specificity 26.2 Human Papillomavirus
for Infection? 26.3 The Respiratory Tract Pathogen Bordetella Binds
23.3 Cathelicidins to Lung Cilia
24.1 Factors That Influence Immunogenicity 26.4 The Common Cold versus Influenza
24.2 ABO Blood Groups: Antigens, Antibodies, 26.5 Intracellular Biofilm Pods Are Reservoirs of Infection
and Karl Landsteiner 26.6 Human Immunodeficiency Virus: Pathogenesis
24.3 Organ Donation and Transplant Rejection 26.7 Spongiform Encephalopathies
24.4 Case Studies in Hypersensitivity 26.8 Atherosclerosis and Coronary Artery Disease
25.1 Finding Virulence Genes: Signature-Tagged 27.1 Antibiotic Spectrum of Activity
Mutagenesis
27.2 Antibiotic Biosynthesis Pathways
25.2 Finding Virulence Genes: In Vivo Expression
27.3 Poking Holes with Nanotubes: A New Antibiotic
Technologies
Therapy
25.3 Caught in the Act: Streptococcus agalactiae Evolved
27.4 Critical Virulence Factors Found in the 1918 Strain
through Conjugation
of Influenza Virus
25.4 Pili Tip Proteins Tighten Their Grip
28.1 API Reactions and Generating a Seven-Digit Microbe
25.5 Normal G-Factor Control of Adenylate Cyclase Identification Code
25.6 Diphtheria Toxin 28.2 DNA-Based Detection Tests
25.7 Identifying New Microbial Toxins 28.3 Microbial Pathogen Detection Gets Wired Up
25.8 Bacterial Covert Operations: Secreted Shigella
Effector Proteins Jam Communications between
Target Cells and Innate Immunity
 PREFACE

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Another random document with
no related content on Scribd:
The Project Gutenberg eBook of Kitty's enemy
This ebook is for the use of anyone anywhere in the United States
and most other parts of the world at no cost and with almost no
restrictions whatsoever. You may copy it, give it away or re-use it
under the terms of the Project Gutenberg License included with this
ebook or online at www.gutenberg.org. If you are not located in the
United States, you will have to check the laws of the country where
you are located before using this eBook.

Title: Kitty's enemy

or, the boy next door.

Author: Eleanora H. Stooke

Release date: September 9, 2023 [eBook #71599]

Language: English

Original publication: London: S. W. Partridge & Co, 1911

*** START OF THE PROJECT GUTENBERG EBOOK KITTY'S

ENEMY ***
Transcriber's note: Unusual and inconsistent spelling is as printed.
MR. DOTTIN SHOOK HIS FIST AT HIS LATE VISITORS,

AND SHUT THE DOOR UPON THEM.

KITTY'S ENEMY:
OR,
 The Boy Next Door.

BY

ELEANORA H. STOOKE.

S. W. PARTRIDGE & CO.

4, 5 & 6 SOHO SQUARE, LONDON, W. I.

 MADE IN GREAT BRITAIN

First published September, 1911

Frequently reprinted

CONTENTS.

Chap.

I. THAT DREADFUL BOY

II. TIM'S REVENGE

III. KITTY INTERVIEWS TIM

IV. MAKING AMENDS

V. A SPOILT PLAN

VI. CONCERNING THE COVERED BASKET

VII. KITTY'S ACCIDENT

VIII. KITTY IS MYSTIFIED

IX. GENERAL BEWILDERMENT

X. SNIP INTRUDES

XI. KITTY ASKS FORGIVENESS OF TIM

XII. TIM ENTERTAINS

XIII. A REGULAR BRICK

XIV. CONCLUSION
 KITTY'S ENEMY.

CHAPTER I.

THAT DREADFUL BOY.

"KITTY, fetch the rake from the tool-house, will you?"

"All right," responded Kitty; and dropping the fork with which she had
been at work moving up the earth in her garden-plot, she hastened
to do her brother's bidding.

Kitty and Bob Glanville were "putting in a morning gardening," as

Bob would have expressed it. They were both enthusiastic
gardeners, and had been allotted a few yards each in the long strip
of ground which stretched at the back of the semi-detached villa in
the small provincial town where they lived. And it being the Easter
holidays—it was at the end of April—they had plenty of time to
devote to tilling their respective garden-plots, and were enjoying their
labours in anticipation of the fine show of flowers they would have
later on from the seeds which they were now sowing. Kitty, who was
a nice-looking little girl of ten years old, with blue eyes and fair, curly
hair, was two years her brother's junior, and being extremely good-
natured she allowed him to order her about, and rarely thought of
refusing to do his will. Bob was very fond of his sister; but he
presumed on his seniority in many ways and expected always to
take the lead.
Kitty having procured the rake returned with it to her brother. "Bob,"
she said, "there's a boy next door—at Mr. Shuttleworth's."

"I know," Bob answered laconically. "I saw him arrive yesterday with
a big portmanteau. He came in a cab with Mr. Shuttleworth."

"Then you may depend Mr. Shuttleworth met him at the station."

Mr. Shuttleworth was an elderly bachelor of studious habits, who

lived next door. Being of a reserved disposition he had little to do
with his neighbours, though sometimes he exchanged a few words
with Mr. and Mrs. Glanville if they happened to be in their front
garden when he was in his, and he always nodded to the children
when he met them. Neither Kitty nor Bob remembered Mr.
Shuttleworth ever having had a visitor before, and they had lived
next door to him for several years now.

"Bob, I wonder who the boy can be," said Kitty, as her brother took
the rake from her hand. "I saw him watching us from Mr.
Shuttleworth's dining-room window. Such a very ugly-looking boy he
is, with red hair, and green goggly eyes, and a snub nose, and a big
mouth. He grinned at me."

"And did you grin back at him?" asked Bob with a laugh.

"No, certainly not," Kitty responded loftily; "as though I should do

that! He's gone now," she added, with a furtive glance at their
neighbour's house. "Oh," she exclaimed in accents of intense
astonishment, a moment later, "why, there he is, staring at us over
the wall! He must be standing on a ladder, I suppose."

Bob paused in the midst of raking his flower-bed smooth and looked
at the wall which divided the two back gardens of the semi-detached
villas. It was a brick wall of about six feet in height, and at the
present minute the head and shoulders of a boy were visible above
it. He had been watching the two young gardeners intently; but when
he perceived they noticed him he looked a trifle embarrassed, and
his freckled countenance deepened in colour.
"Yes, he must be on a ladder," Bob agreed. "The idea of his spying
on us like that! I shall take no notice of him!"

"Nor I," said Kitty; and she turned her attention to her garden again;
but she could not refrain from glancing at the partition wall frequently,
and every time she did so there was still the ugly boy watching them.

"Do you think he wishes to speak to us?" she suggested to her

brother at length. "Shall we ask him if he wants anything?"

"Most certainly not," Bob responded. "I call it cheek his perching
himself there; if I spoke to him I should tell him so."

Meanwhile the boy next door—Tim Shuttleworth—who had come out

in hopes of picking an acquaintance with his neighbours, saw that,
though they had observed him, they had no intention of speaking to
him, and was consequently disappointed. Although he had only
arrived on a visit to his uncle, Mr. Shuttleworth, on the preceding day,
time was hanging heavily on his hands already. At breakfast his
uncle had told him he might go where he pleased and do exactly as
he liked, and had remarked that no doubt he would find his own
amusements—adding that there were young folks in the next house
he could play with, but he had evidently not thought of introducing
him to them. And then he had shut himself up in his study, leaving
his nephew to his own devices.

From the dining-room window Tim had taken stock of the next door
garden, and of Kitty and of Bob Glanville with their lemon-and-white
fox-terrier, Snip, in close attendance upon them; and seeing that Bob
was about his age and that Kitty looked a merry, nice little girl, he
had thought how pleasant it would be for him to know them.
Accordingly he had procured a short ladder, which he had found
hanging against the tool-house wall, and had taken up a position
where he could overlook the next door garden. He had seen Kitty
call her brother's attention to him, and had observed them
whispering together, and he deemed it almost unsociable for them
not to speak.
By-and-by Snip trotted up the garden path nearest the partition wall,
and suddenly paused, sniffing the air in a suspicious manner. Then
he looked up and caught sight of the stranger; but instead of barking
at him, he seated himself on his haunches and contemplated him
with a world of perplexity in his brown eyes, into which there
gradually crept the loveliest possible canine smile.

"Good doggie," Tim murmured. "You're a jolly, friendly little chap."

And Snip wagged his tail and wriggled with excitement, every nerve
in his body on the twitch, for he liked the voice which addressed him,
and he liked the countenance of the next door boy, which did not
seem ugly to him at all.

"Look at Snip," said Kitty in an undertone. "The boy's speaking to

him, and he's quite pleased."

"Little sniveller!" exclaimed Bob. He immediately called to the dog to

come away; whereupon Tim made a grimace at him and descended
the ladder in haste.

"What an impertinent boy!" cried Kitty. "But I don't think it was nice of
you to call Snip away like that; he was doing no harm, and it seemed
rather rude, I thought. I know you didn't mean it to be so," she added
hastily, noticing that her brother's colour had risen.

"The fellow had no right there at all," Bob replied. "If any one was
rude, he was. He is evidently full of cheek. Did you see what a face
he made at me?"

"Yes, and didn't he look ugly?" cried Kitty, smiling at the

remembrance of the distorted countenance which had disappeared
so suddenly on the other side of the wall. "I wonder if he is a relation
of Mr. Shuttleworth's, and if he will stay long?"

"You'd like to know all about him, wouldn't you, Kitty?" her brother
said, laughing. "You wouldn't be a girl if you weren't curious."

"Boys are curious, too, only they never will own it," Kitty retorted.
"Some are," Bob admitted. "That red-headed chap is, I should think,
or he wouldn't have looked at us over the wall like that."

Tim Shuttleworth watched the young folk next door constantly during
the next day or two, for they spent much time in the back garden; but
he always kept behind the lace curtains in the dining-room, and did
not try to attract their notice again. Their doings interested him
exceedingly. Sometimes they would place Snip on the wall to watch
for cats, and on one occasion the little dog hunted one of his natural
enemies all about Mr. Shuttleworth's garden, around the house, and
out into the road. No one would have been more shocked and
horrified than the two young Glanvilles had Snip really caught a cat;
but he had never done so, and it was most unlikely he ever would,
so his mistress and master saw no harm in permitting him to enjoy
his little game, though they had several times been told by their
father not to encourage him in his favourite pursuit.

Tim knew his visit to his uncle was to be of long duration. He was the
eldest of several children, and as his father was only a clerk in a
bank in Dublin, his home was a comparatively poor one. During the
winter he had had a serious attack of pneumonia, and when his
uncle had written and suggested that he should spend the summer
with him in England, at the small country town where Mr.
Shuttleworth had lived for many years, his parents had promptly
accepted the invitation; for the doctor who had attended him during
his illness had declared that a thorough change of air and scene,
and an absence from school for a few months, would be to his
ultimate benefit. Tim had known but little of his uncle up to the
present, having only seen him on the rare occasions when he had
come on a visit to Dublin, and when he found out how thoroughly
engrossed his relative was in his books and his studies, he was
dismayed at the prospect of the months of loneliness, he feared he
must look forward to. If only the children next door would seek to
know him! But he realized that they were not prepossessed with his
appearance, and he was too shy and too proud to make any
overtures of friendship to them, and determined to find his own
amusements, as his uncle expected him to do.
Accordingly, he selected a sunny spot in the back garden—which
was almost a wilderness, for Mr. Shuttleworth did not care for
flowers, and bought his vegetables—and began to turn up the soil. It
was hard work, for the ground had long been neglected, and the
tools he found in the tool-house were blunt and heavy; but he
persevered. He would have a garden of his own, he decided, which
should rival those of the children next door; and at length he
succeeded in getting the earth in fairly good condition, and planted it
with the seeds of nasturtiums and mignonette and other hardy
annuals which he purchased with his own pocket-money.

There came a night when Tim went to bed greatly pleased with the
result of his finished labours; but the next morning he found they had
been all in vain, for on going into the back garden after breakfast, he
discovered that some animal—a dog, presumably—had been
scratching and digging holes in the ground he had prepared so
carefully for the reception of the seeds.

His indignation was intense; and fetching the ladder from the tool-
house, he placed it against the partition wall, climbed it, and looked
into the next garden. There were the sister and brother, their
attention fixed on the contents of a big box which they had placed
against the wall, and near them was Snip, his nose covered with
earth.

Tim's wrath burst bounds as he regarded the trio, and he poured

forth a volume of angry words, his Irish accent growing more and
more pronounced as he proceeded, in which he accused Kitty and
Bob of having deliberately put Snip in over the wall to destroy his
garden. He vowed vengeance upon them, and at last, overcome with
rage, he shook his fist at them and then abruptly quitted the scene.

"What a dreadful boy!" cried Kitty, who had turned quite pale. "What
can he mean by speaking to us like that? What can Snip have
done?"

"Destroyed his garden, he said," responded Bob. "I didn't put Snip
over the wall, though. If he's been in there, he went by himself. My,
what a temper that fellow has! I believe, as you say, he's a dreadful
boy."

CHAPTER II.
TIM'S REVENGE.

A SECOND survey of his garden did not lessen Tim Shuttleworth's

indignation in the least; and too angry to endeavour to repair the
mischief done, he marched into the house and took up his position
behind the lace curtains in the dining-room, through which he kept a
watchful eye on the Glanvilles' back garden, his mind full of wild
plans for revenge.

What could there be in the box which had been placed against the
partition wall, Tim pondered? It had not been there yesterday; but
that it contained something very attractive was evident, for Mr. and
Mrs. Glanville, and the cook, and housemaid, and later the boot boy,
all visited it one by one.

"I expect they've been given some potting plants," Tim decided at
length. "Yes, that must be it, for they've put the box against the
sunny wall. How would they like it, I wonder, if someone else's dog
came into their garden and dug big holes in their flower-beds? I've a
great mind to make a complaint against them to uncle; but no, I won't
be a sneak. I'll find some way of paying them out myself."

Later in the morning—which was cold and windy for the end of April
—Tim went for a stroll in the town, and as he walked up the main
street feeling very lonely, and longing for congenial society, he
encountered Kitty and Bob coming out of a sweet shop, followed by
Snip. He gave them one swift glance in passing, and noticed that
Kitty, who had turned rosy red, was pretending not to see him, and
that Bob was regarding him with eyes which twinkled humorously. It
flashed upon him that his exhibition of temper had caused the other
boy amusement, and the knowledge only made him more angry. He
felt it was adding insult to injury to laugh at him; and when Snip,
evidently recognising him, obtruded himself on his notice by jumping
against him, he gruffly told him to get away.

"Come here, Snip," commanded Bob, in a voice which betrayed

merriment. "Come here, sir, im—mediately!"

So Snip, looking decidedly crestfallen, turned his attention once

more to his young mistress and master; and Tim went on, his head
held haughtily, his hands thrust deep into his trousers' pockets.

"Don't you think we ought to have told him that it was not our fault
that Snip did harm to his garden?" asked Kitty of her brother,
glancing over her shoulder at Tim's retreating form. She spoke rather
indistinctly, because her mouth was full of sweets.

"What! After the way he spoke to us?" cried Bob. "Not likely. Let the
fellow think what he likes."

"It doesn't seem right to let him think what isn't true, though, does it?"

"I don't see that it matters."

"No, I don't know that it does." Kitty agreed, as usual coming round
to her brother's way of reasoning. "How crossly he spoke to Snip,
didn't he? Poor little Snip, he meant no harm."

Meanwhile, Tim having reached the corner of the street, looked back
and saw that the Glanvilles were not going home, but had turned
down a side road leading in the opposite direction. A sudden,
malicious thought flashed through his mind, which caused him to
turn and hastily retrace his footsteps. On his arrival at his uncle's
house he went immediately to the back garden, and, armed with a
spade, he mounted the ladder, which he had left against the wall,
and peered cautiously around the Glanville's garden. No one was in
sight, and, being assured that he was unobserved, Tim leaned over
the wall, the better to view the box beneath; and then, with the aid of
the spade, he tried to lift the lid, but failed in the attempt. However, it
did not matter what was inside, he told himself; if it was something
breakable so much the better.

For a few minutes Tim hesitated, his conscience telling him he was
about to act very wrongly; but the remembrance of the grievance, he
believed he had every right to cherish against Kitty and Bob nerved
him to do that which he afterwards bitterly repented having done.
And, with a vigorous shove from the spade, he overturned the box
with a jerk, descended the ladder hastily, and hurried back to the
house, his feelings those of mingled fear and exultation—fear lest he
might have been seen after all, and exultation because he believed
he had now scored off the next door children.

When uncle and nephew met at the mid-day meal that day, it
occurred to Mr. Shuttleworth to ask Tim how he had spent the
morning.

"Oh, I've been in town," Tim answered, flushing, "and—and in the

garden. I don't think much of the shops here, Uncle John; they're not
to be compared with ours in Dublin."

"No, of course not. You must remember this is only a small country
town—not to be compared in any way to 'Dublin's fair city,'" Mr.
Shuttleworth replied with a smile. He was a tall man with stooping
shoulders, and near-sighted eyes which peered at Tim very kindly
through spectacles, and he was very clever; but with all his
cleverness, he did not understand children, which was a pity for Tim.
"Have you spoken to the young folks next door yet?" he inquired.

"Oh, yes," assented Tim, trying to respond in a careless tone. "I've

spoken to them; but I don't think much of them, Uncle John."
"Dear me. Why not?" There was surprise in Mr. Shuttleworth's voice.

"They're stuck-up," Tim asserted, after a moment's reflection.

"Stuck-up? You astonish me. I don't know much of my neighbours,

but Mr. and Mrs. Glanville seem pleasant people to talk to, and the
children—though I have never spoken to them—always struck me as
being well behaved. They go to school, I believe; but I suppose it is
holiday-time now. Don't you think you'll be friendly with them, then?"

Tim shook his head decidedly, and his uncle dropped the subject,
telling himself that perhaps his nephew, like himself, preferred his
own company.

As soon as Mr. Shuttleworth had returned to his study, and the

servant had cleared the table, Tim took up his post at the window, on
the alert to see what would happen when Kitty and Bob paid their
next visit to the back garden. It was less cold than it had been earlier
in the day, and the wind had gone down, so he opened the window,
and it was not long before he heard voices, and the sister and
brother appeared, the former carrying something very carefully in a
saucer.

Tim's heart beat unevenly as he watched the children. As he had

anticipated, they made straight for the box, and great was the
consternation depicted on their faces when they found it had been
overturned.

"Oh, Bob, what can have happened?" cried Kitty; and the listener at
the window caught a note of alarm in her voice. "Oh, dear! Oh, dear!"

"Don't be frightened," Bob answered reassuringly. "It's only upset; I'll

soon put it right. I hope a dog hasn't been here; it wouldn't have
been Snip, anyway, for we warned him off, and he quite understood.
Oh, be careful, Kitty! You're spilling the bread and milk all over your
frock."