Microbiology, 2021

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58 2 • How We See the Invisible World Figure 2.29 Society for <strong>Microbiology</strong>) (credit “TEM”: modification of work by American Society for <strong>Microbiology</strong>; credit “SEM”: modification of work by American Figure 2.30 2.4 Staining Microscopic Specimens Learning Objectives By the end of this section, you will be able to: • Differentiate between simple and differential stains • Describe the unique features of commonly used stains • Explain the procedures and name clinical applications for Gram, endospore, acid-fast, negative capsule, and flagella staining In their natural state, most of the cells and microorganisms that we observe under the microscope lack color and contrast. This makes it difficult, if not impossible, to detect important cellular structures and their distinguishing characteristics without artificially treating specimens. We have already alluded to certain Access for free at openstax.org.
2.4 • Staining Microscopic Specimens 59 techniques involving stains and fluorescent dyes, and in this section we will discuss specific techniques for sample preparation in greater detail. Indeed, numerous methods have been developed to identify specific microbes, cellular structures, DNA sequences, or indicators of infection in tissue samples, under the microscope. Here, we will focus on the most clinically relevant techniques. Preparing Specimens for Light Microscopy In clinical settings, light microscopes are the most commonly used microscopes. There are two basic types of preparation used to view specimens with a light microscope: wet mounts and fixed specimens. The simplest type of preparation is the wet mount, in which the specimen is placed on the slide in a drop of liquid. Some specimens, such as a drop of urine, are already in a liquid form and can be deposited on the slide using a dropper. Solid specimens, such as a skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid used is simply water, but often stains are added to enhance contrast. Once the liquid has been added to the slide, a coverslip is placed on top and the specimen is ready for examination under the microscope. The second method of preparing specimens for light microscopy is fixation. The “fixing” of a sample refers to the process of attaching cells to a slide. Fixation is often achieved either by heating (heat fixing) or chemically treating the specimen. In addition to attaching the specimen to the slide, fixation also kills microorganisms in the specimen, stopping their movement and metabolism while preserving the integrity of their cellular components for observation. To heat-fix a sample, a thin layer of the specimen is spread on the slide (called a smear), and the slide is then briefly heated over a heat source (Figure 2.31). Chemical fixatives are often preferable to heat for tissue specimens. Chemical agents such as acetic acid, ethanol, methanol, formaldehyde (formalin), and glutaraldehyde can denature proteins, stop biochemical reactions, and stabilize cell structures in tissue samples (Figure 2.31). Figure 2.31 (a) A specimen can be heat-fixed by using a slide warmer like this one. (b) Another method for heat-fixing a specimen is to hold a slide with a smear over a microincinerator. (c) This tissue sample is being fixed in a solution of formalin (also known as formaldehyde). Chemical fixation kills microorganisms in the specimen, stopping degradation of the tissues and preserving their structure so that they can be examined later under the microscope. (credit a: modification of work by Nina Parker; credit b: modification of work by Nina Parker; credit c: modification of work by “University of Bristol”/YouTube) In addition to fixation, staining is almost always applied to color certain features of a specimen before examining it under a light microscope. Stains, or dyes, contain salts made up of a positive ion and a negative ion. Depending on the type of dye, the positive or the negative ion may be the chromophore (the colored ion); the other, uncolored ion is called the counterion. If the chromophore is the positively charged ion, the stain is classified as a basic dye; if the negative ion is the chromophore, the stain is considered an acidic dye. Dyes are selected for staining based on the chemical properties of the dye and the specimen being observed, which determine how the dye will interact with the specimen. In most cases, it is preferable to use a positive stain, a dye that will be absorbed by the cells or organisms being observed, adding color to objects of interest to make them stand out against the background. However, there are scenarios in which it is advantageous to use a negative stain, which is absorbed by the background but not by the cells or organisms in the specimen. Negative staining produces an outline or silhouette of the organisms against a colorful background (Figure
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Microbiology SENIOR CONTRIBUTING AU
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OPENSTAX OpenStax provides free, pe
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Contents Preface 1 CHAPTER 1 An Inv
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9.2 Oxygen Requirements for Microbi
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18.1 Overview of Specific Adaptive
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Appendix C Metabolic Pathways 1155
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Preface 1 PREFACE Welcome to Microb
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Preface 3 that often confer critica
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Preface 5 further. Our features inc
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Page 23 and 24: CHAPTER 1 An Invisible World Figure
Page 25 and 26: 1.1 • What Our Ancestors Knew 11
Page 31 and 32: 1.2 • A Systematic Approach 17 Th
Page 33 and 34: 1.2 • A Systematic Approach 19 Fi
Page 35 and 36: 1.2 • A Systematic Approach 21 Ta
Page 37 and 38: 1.3 • Types of Microorganisms 23
Page 45 and 46: 1 • Summary 31 SUMMARY 1.1 What O
Page 47 and 48: 1 • Review Questions 33 17. The p
Page 49 and 50: CHAPTER 2 How We See the Invisible
Page 51 and 52: 2.1 • The Properties of Light 37
Page 53 and 54: 2.1 • The Properties of Light 39
Page 55 and 56: 2.2 • Peering Into the Invisible
Page 57 and 58: 2.3 • Instruments of Microscopy 4
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Page 75 and 76: 2.4 • Staining Microscopic Specim
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Page 87 and 88: CHAPTER 3 The Cell Figure 3.1 Micro
Page 89 and 90: 3.1 • Spontaneous Generation 75 F
Page 91 and 92: 3.2 • Foundations of Modern Cell
Page 99 and 100: 3.3 • Unique Characteristics of P
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3.4 • Unique Characteristics of E
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3 • Summary 125 SUMMARY 3.1 Spont
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3 • Review Questions 127 3. Which
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3 • Review Questions 129 Critical
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CHAPTER 4 Prokaryotic Diversity Fig
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4.1 • Prokaryote Habitats, Relati
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4.2 • Proteobacteria 139 for or s
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4.2 • Proteobacteria 141 Class Al
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4.2 • Proteobacteria 143 Class Be
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4.2 • Proteobacteria 145 Figure 4
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4.2 • Proteobacteria 147 Class Ga
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4.3 • Nonproteobacteria Gram-Nega
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4.4 • Gram-Positive Bacteria 157
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4.6 • Archaea 167 The class Therm
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4.6 • Archaea 169 thus is a livin
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4 • Summary 171 SUMMARY 4.1 Proka
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4 • Review Questions 173 REVIEW Q
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4 • Review Questions 175 39. Expl
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CHAPTER 5 The Eukaryotes of Microbi
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5.1 • Unicellular Eukaryotic Para
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Figure 5.7 5.1 • Unicellular Euka
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5.2 • Parasitic Helminths 193 hel
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5.2 • Parasitic Helminths 195 Fig
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5.2 • Parasitic Helminths 197 Fig
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5.2 • Parasitic Helminths 199 MIC
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5.3 • Fungi 201 Figure 5.25 Multi
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5.3 • Fungi 203 Figure 5.27 zygos
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5.3 • Fungi 205 diploid stages (F
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5.3 • Fungi 207 Figure 5.32 prese
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5.3 • Fungi 209 MICRO CONNECTIONS
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5.4 • Algae 211 and other photosy
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5.5 • Lichens 213 Figure 5.37 Chl
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5.5 • Lichens 215 marina. (b) Thi
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5 • Review Questions 217 REVIEW Q
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CHAPTER 6 Acellular Pathogens Figur
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6.1 • Viruses 221 assembly of vir
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6.1 • Viruses 223 Viral Structure
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6.1 • Viruses 225 Figure 6.5 (a)
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6.1 • Viruses 227 LINK TO LEARNIN
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6.2 • The Viral Life Cycle 229 on
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6.2 • The Viral Life Cycle 231 Fi
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6.2 • The Viral Life Cycle 237 by
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6.2 • The Viral Life Cycle 239 Ca
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6.3 • Isolation, Culture, and Ide
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6.4 • Viroids, Virusoids, and Pri
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6.4 • Viroids, Virusoids, and Pri
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6 • Summary 253 SUMMARY 6.1 Virus
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6 • Review Questions 255 18. A/an
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CHAPTER 7 Microbial Biochemistry Fi
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7.1 • Organic Molecules 259 Figur
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7.1 • Organic Molecules 261 Figur
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7.2 • Carbohydrates 263 and the m
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7.2 • Carbohydrates 265 Figure 7.
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7.3 • Lipids 267 Figure 7.11 Star
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7.3 • Lipids 269 Figure 7.13 This
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7.3 • Lipids 271 Figure 7.15 Five
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7.4 • Proteins 273 Figure 7.17 Am
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7.4 • Proteins 275 Figure 7.19 Re
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7.4 • Proteins 277 Figure 7.23 Pr
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7.5 • Using Biochemistry to Ident
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7.5 • Using Biochemistry to Ident
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7 • Review Questions 283 as hormo
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7 • Review Questions 285 19. A tr
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CHAPTER 8 Microbial Metabolism Figu
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8.1 • Energy, Matter, and Enzymes
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8.2 • Catabolism of Carbohydrates
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8.3 • Cellular Respiration 301 8.
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8.3 • Cellular Respiration 303 Fi
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8.4 • Fermentation 305 If respira
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8.4 • Fermentation 307 Common Fer
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8.5 • Catabolism of Lipids and Pr
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8.6 • Photosynthesis 311 independ
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8.6 • Photosynthesis 313 Oxygenic
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8.7 • Biogeochemical Cycles 315 L
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8.7 • Biogeochemical Cycles 317 N
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8.7 • Biogeochemical Cycles 319 F
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8.7 • Biogeochemical Cycles 321 F
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8 • Summary 323 oxygen as the fin
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8 • Review Questions 325 4. To wh
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8 • Review Questions 327 Matching
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CHAPTER 9 Microbial Growth Figure 9
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9.1 • How Microbes Grow 331 and a
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9.1 • How Microbes Grow 333 The G
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9.1 • How Microbes Grow 335 Susta
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9.1 • How Microbes Grow 337 chang
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9.1 • How Microbes Grow 339 Figur
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9.1 • How Microbes Grow 341 trans
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9.1 • How Microbes Grow 343 micro
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9.2 • Oxygen Requirements for Mic
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9.3 • The Effects of pH on Microb
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9.4 • Temperature and Microbial G
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9.4 • Temperature and Microbial G
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9.5 • Other Environmental Conditi
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9.6 • Media Used for Bacterial Gr
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9 • Summary 361 SUMMARY 9.1 How M
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9 • Review Questions 363 7. Filam
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9 • Review Questions 365 Matching
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9 • Review Questions 367 Critical
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CHAPTER 10 Biochemistry of the Geno
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10.1 • Using Microbiology to Disc
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10.2 • Structure and Function of
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10 • Summary 401 SUMMARY 10.1 Usi
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10 • Review Questions 403 3. Whic
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10 • Review Questions 405 Short A
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CHAPTER 11 Mechanisms of Microbial
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11.1 • The Functions of Genetic M
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11.2 • DNA Replication 411 Figure
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11.2 • DNA Replication 413 Figure
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11.2 • DNA Replication 415 strand
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11.2 • DNA Replication 417 telome
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11.3 • RNA Transcription 419 Figu
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11.3 • RNA Transcription 421 the
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11.4 • Protein Synthesis (Transla
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11.5 • Mutations 429 Effects of M
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11.5 • Mutations 431 In recent ye
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11.5 • Mutations 433 Figure 11.20
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11.5 • Mutations 435 formation of
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11.6 • How Asexual Prokaryotes Ac
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11.7 • Gene Regulation: Operon Th
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11 • Summary 457 SUMMARY 11.1 The
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11 • Summary 459 undamaged DNA st
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11 • Review Questions 461 11. Whi
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11 • Review Questions 463 46. ___
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71. The following figure is from Mo
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CHAPTER 12 Modern Applications of M
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12.1 • Microbes and the Tools of
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12.2 • Visualizing and Characteri
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12.3 • Whole Genome Methods and P
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12.3 • Whole Genome Methods and P
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12.4 • Gene Therapy 499 Figure 12
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12.4 • Gene Therapy 501 overall w
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12 • Summary 503 SUMMARY 12.1 Mic
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12 • Review Questions 505 11. The
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CHAPTER 13 Control of Microbial Gro
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13.1 • Controlling Microbial Grow
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13.2 • Using Physical Methods to
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13.3 • Using Chemicals to Control
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13.4 • Testing the Effectiveness
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13 • Summary 553 commonly used to
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13 • Review Questions 555 12. Ble
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CHAPTER 14 Antimicrobial Drugs Figu
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14.1 • History of Chemotherapy an
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14.1 • History of Chemotherapy an
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14.2 • Fundamentals of Antimicrob
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14.2 • Fundamentals of Antimicrob
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14.3 • Mechanisms of Antibacteria
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14.4 • Mechanisms of Other Antimi
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14.5 • Drug Resistance 591 Common
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14.5 • Drug Resistance 593 negati
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14.5 • Drug Resistance 595 Clavul
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14.7 • Current Strategies for Ant
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14.7 • Current Strategies for Ant
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14 • Summary 605 cells. • Becau
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14 • Review Questions 609 50. Why
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CHAPTER 15 Microbial Mechanisms of
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15.1 • Characteristics of Infecti
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15.2 • How Pathogens Cause Diseas
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15.3 • Virulence Factors of Bacte
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15.4 • Virulence Factors of Eukar
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15.4 • Virulence Factors of Eukar
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15 • Review Questions 647 protect
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15 • Review Questions 649 Critica
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CHAPTER 16 Disease and Epidemiology
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16.1 • The Language of Epidemiolo
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16.1 • The Language of Epidemiolo
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16.2 • Tracking Infectious Diseas
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16.3 • Modes of Disease Transmiss
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16.4 • Global Public Health 673 C
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16.4 • Global Public Health 675 S
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16 • Summary 677 SUMMARY 16.1 The
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16 • Review Questions 679 Matchin
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16 • Review Questions 681 20. Wha
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CHAPTER 17 Innate Nonspecific Host
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17.1 • Physical Defenses 685 Over
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17.1 • Physical Defenses 687 know
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17.1 • Physical Defenses 689 Figu
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17.2 • Chemical Defenses 691 Phys
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17.2 • Chemical Defenses 693 sali
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17.2 • Chemical Defenses 695 prod
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17.2 • Chemical Defenses 697 Figu
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17.2 • Chemical Defenses 699 Clin
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17.3 • Cellular Defenses 701 Hema
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17.3 • Cellular Defenses 703 stra
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17.3 • Cellular Defenses 705 Clin
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17.3 • Cellular Defenses 707 Figu
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17.4 • Pathogen Recognition and P
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17.4 • Pathogen Recognition and P
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17.5 • Inflammation and Fever 713
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17 • Summary 719 SUMMARY 17.1 Phy
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17 • Review Questions 721 8. Hist
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17 • Review Questions 723 Short A
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CHAPTER 18 Adaptive Specific Host D
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18.1 • Overview of Specific Adapt
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18.2 • Major Histocompatibility C
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18.3 • T Lymphocytes and Cellular
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18.4 • B Lymphocytes and Humoral
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18.4 • B Lymphocytes and Humoral
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18.5 • Vaccines 751 pathogen—bu
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18.5 • Vaccines 753 Eye on Ethics
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18.5 • Vaccines 755 for Disease C
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18.5 • Vaccines 757 Classes of Va
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18 • Summary 759 SUMMARY 18.1 Ove
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18 • Review Questions 761 5. MHC
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18 • Review Questions 763 20. Mat
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CHAPTER 19 Diseases of the Immune S
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19.1 • Hypersensitivities 767 Fig
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19.1 • Hypersensitivities 769 bin
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19.1 • Hypersensitivities 771 mec
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19.1 • Hypersensitivities 775 CHE
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19.1 • Hypersensitivities 777 Cli
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19.1 • Hypersensitivities 779 MIC
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19.2 • Autoimmune Disorders 783 t
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19.2 • Autoimmune Disorders 785 F
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19.2 • Autoimmune Disorders 787 M
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19.2 • Autoimmune Disorders 789 F
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19.3 • Organ Transplantation and
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19.4 • Immunodeficiency 793 and m
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19.4 • Immunodeficiency 795 CHECK
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19.5 • Cancer Immunobiology and I
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19 • Summary 799 SUMMARY 19.1 Hyp
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19 • Review Questions 801 13. All
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CHAPTER 20 Laboratory Analysis of t
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20.1 • Polyclonal and Monoclonal
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20.2 • Detecting Antigen-Antibody
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20.3 • Agglutination Assays 823 a
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20.3 • Agglutination Assays 825 F
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20.3 • Agglutination Assays 827 e
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20.3 • Agglutination Assays 829 s
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20.4 • EIAs and ELISAs 831 Mechan
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20.4 • EIAs and ELISAs 833 Figure
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20.4 • EIAs and ELISAs 837 • Wh
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20.5 • Fluorescent Antibody Techn
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20 • Review Questions 849 20.4 EI
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20 • Review Questions 851 15. Sup
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CHAPTER 21 Skin and Eye Infections
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21.1 • Anatomy and Normal Microbi
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21.2 • Bacterial Infections of th
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21.3 • Viral Infections of the Sk
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21.3 • Viral Infections of the Sk
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21.4 • Mycoses of the Skin 881 se
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21.4 • Mycoses of the Skin 883 fr
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21.4 • Mycoses of the Skin 885 Fi
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21.5 • Protozoan and Helminthic I
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21.5 • Protozoan and Helminthic I
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21 • Summary 891 SUMMARY 21.1 Ana
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CHAPTER 22 Respiratory System Infec
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22.3 • Viral Infections of the Re
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22.4 • Respiratory Mycoses 931 ha
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22.4 • Respiratory Mycoses 933 Fi
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22.4 • Respiratory Mycoses 935 re
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Figure 22.29 22.4 • Respiratory M
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22 • Review Questions 939 200 vir
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CHAPTER 23 Urogenital System Infect
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23.5 • Fungal Infections of the R
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23.6 • Protozoan Infections of th
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23 • Summary 975 SUMMARY 23.1 Ana
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CHAPTER 24 Digestive System Infecti
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24.2 • Microbial Diseases of the
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24.4 • Viral Infections of the Ga
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24.6 • Helminthic Infections of t
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24 • Summary 1033 SUMMARY 24.1 An
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CHAPTER 25 Circulatory and Lymphati
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25.1 • Anatomy of the Circulatory
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25.1 • Anatomy of the Circulatory
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25.3 • Viral Infections of the Ci
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Figure 25.26 25.3 • Viral Infecti
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25.4 • Parasitic Infections of th
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25 • Review Questions 1089 • To
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25 • Review Questions 1091 Critic
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CHAPTER 26 Nervous System Infection
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26.1 • Anatomy of the Nervous Sys
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26.1 • Anatomy of the Nervous Sys
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26.2 • Bacterial Diseases of the
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26.3 • Acellular Diseases of the
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Figure 26.19 26.3 • Acellular Dis
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26.4 • Fungal and Parasitic Disea
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26 • Review Questions 1133 are fa
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26 • Review Questions 1135 Matchi
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26 • Review Questions 1137 59. Th
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A • Fundamentals of Physics and C
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B • Mathematical Basics 1149 APPE
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B • Mathematical Basics 1151 Mult
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B • Mathematical Basics 1153 The
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C • Metabolic Pathways 1155 APPEN
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C • Metabolic Pathways 1157 Entne
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C • Metabolic Pathways 1159 Figur
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C • Metabolic Pathways 1161 Elect
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APPENDIX D Taxonomy of Clinically R
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D • Taxonomy of Clinically Releva
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E • Glossary 1177 APPENDIX E Glos
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E • Glossary 1179 destruction by
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E • Glossary 1181 pollutants from
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E • Glossary 1183 chromosome disc
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E • Glossary 1185 cysts are forme
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E • Glossary 1187 occurring ectop
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E • Glossary 1189 molecules of DN
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E • Glossary 1191 glycoprotein co
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E • Glossary 1193 image point is
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E • Glossary 1195 discontinuously
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E • Glossary 1197 infection cause
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E • Glossary 1199 target cells by
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E • Glossary 1201 intact ribosome
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E • Glossary 1203 point source sp
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E • Glossary 1205 complexes forme
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E • Glossary 1207 antibody is fir
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E • Glossary 1209 the number of c
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E • Glossary 1211 the reciprocal
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E • Glossary 1213 vector animal (
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1215 ANSWER KEY Chapter 1 1. D 2. D
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1217 B 18. A, C, B 19. peristalsis
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Index 1219 INDEX Symbols +ssRNA 237
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Index 1221 antigen-presenting cells
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Index 1223 Bordetella 142 Bordetell
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Index 1225 chromosomes 372, 394 chr
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Index 1227 de Duve 113 dead zone 35
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Index 1229 Enteroinvasive E. coli (
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Index 1231 Gardnerella vaginalis 47
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Index 1233 shunt 298 Hfr cell 443 H
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Index 1235 iron lungs 1118 iron-sul
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Index 1237 maturation 230 mature na
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Index 1239 Health 501 National Noti
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Index 1241 patterns (PAMPs) 710 pat
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Index 1243 primary lymphoid tissue
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Index 1245 596, 1048, 1051, 1051 ri
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Index 1247 sterilant 511, 541 steri
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Index 1249 toxigenicity 633 toxin 6
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Index 1251 water activity 522 water
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Microbiology, 2021

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