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840 20 • Laboratory Analysis of the Immune Response Immunoblots & Enzyme Immunoassays Type of Assay Mechanism Specific Procedures Examples Immunoblots Uses enzyme-antibody conjugates to identify specific proteins that have been transferred to an absorbent membrane Western blot: Detects the presence of a particular protein Detecting the presence of HIV peptides (or peptides from other infectious agents) in patient sera Immunostaining Uses enzyme-antibody conjugates to stain specific molecules on or in cells Immunohistochemistry: Used to stain specific cells in a tissue Stain for presence of CD8 cells in host tissue Enzyme-linked immunosorbent assay (ELISA) Uses enzyme-antibody conjugates to quantify target molecules Direct ELISA: Uses a single antibody to detect the presence of an antigen Indirect ELISA: Measures the amount of antibody produced against an antigen Detection of HIV antigen p24 up to one month after being infected Detection of HIV antibodies in serum Immunochromatographic (lateral flow) assays Techniques use the capture of flowing, colorlabeled antigen-antibody complexes by fixed antibody for disease diagnosis Sandwich ELISA: Measures the amount of antigen bound by the antibody Detection of antibodies for various pathogens in patient sera (e.g., rapid strep, malaria dipstick) Pregnancy test detecting human chorionic gonadotrophin in urine Table 20.4 Clinical Focus Part 3 Although the indirect ELISA for HIV is a sensitive assay, there are several complicating considerations. First, if an infected person is tested too soon after becoming infected, the test can yield false-negative results. The seroconversion window is generally about three weeks, but in some cases, it can be more than two months. Access for free at openstax.org.
20.5 • Fluorescent Antibody Techniques 841 In addition to false negatives, false positives can also occur, usually due to previous infections with other viruses that induce cross-reacting antibodies. The false-positive rate depends on the particular brand of test used, but 0.5% is not unusual. 10 Because of the possibility of a false positive, all positive tests are followed up with a confirmatory test. This confirmatory test is often an immunoblot (western blot) in which HIV peptides from the patient’s blood are identified using an HIV-specific mAb-enzyme conjugate. A positive western blot would confirm an HIV infection and a negative blot would confirm the absence of HIV despite the positive ELISA. Unfortunately, western blots for HIV antigens often yield indeterminant results, in which case, they neither confirm nor invalidate the results of the indirect ELISA. In fact, the rate of indeterminants can be 10–49% (which is why, combined with their cost, western blots are not used for screening). Similar to the indirect ELISA, an indeterminant western blot can occur because of cross-reactivity or previous viral infections, vaccinations, or autoimmune diseases. • Of the 1300 patients being tested, how many false-positive ELISA tests would be expected? • Of the false positives, how many indeterminant western blots could be expected? • How would the hospital address any cases in which a patient’s western blot was indeterminant? Jump to the previous Clinical Focus box. Jump to the next Clinical Focus box. 20.5 Fluorescent Antibody Techniques Learning Objectives By the end of this section, you will be able to: • Describe the benefits of immunofluorescent antibody assays in comparison to nonfluorescent assays • Compare direct and indirect fluorescent antibody assays • Explain how a flow cytometer can be used to quantify specific subsets of cells present in a complex mixture of cell types • Explain how a fluorescence-activated cell sorter can be used to separate unique types of cells Rapid visualization of bacteria from a clinical sample such as a throat swab or sputum can be achieved through fluorescent antibody (FA) techniques that attach a fluorescent marker (fluorogen) to the constant region of an antibody, resulting in a reporter molecule that is quick to use, easy to see or measure, and able to bind to target markers with high specificity. We can also label cells, allowing us to precisely quantify particular subsets of cells or even purify these subsets for further research. As with the enzyme assays, FA methods may be direct, in which a labeled mAb binds an antigen, or indirect, in which secondary polyclonal antibodies bind patient antibodies that react to a prepared antigen. Applications of these two methods were demonstrated in Figure 2.19. FA methods are also used in automated cell counting and sorting systems to enumerate or segregate labeled subpopulations of cells in a sample. Direct Fluorescent Antibody Techniques Direct fluorescent antibody (DFA) tests use a fluorescently labeled mAb to bind and illuminate a target antigen. DFA tests are particularly useful for the rapid diagnosis of bacterial diseases. For example, fluorescence-labeled antibodies against Streptococcus pyogenes (group A strep) can be used to obtain a diagnosis of strep throat from a throat swab. The diagnosis is ready in a matter of minutes, and the patient can be started on antibiotics before even leaving the clinic. DFA techniques may also be used to diagnose pneumonia caused by Mycoplasma pneumoniae or Legionella pneumophila from sputum samples (Figure 20.28). The fluorescent antibodies bind to the bacteria on a microscope slide, allowing ready detection of the bacteria using a fluorescence microscope. Thus, the DFA technique is valuable for visualizing certain bacteria that are difficult to isolate or culture from patient samples. 10 Thomas, Justin G., Victor Jaffe, Judith Shaffer, and Jose Abreu, “HIV Testing: US Recommendations 2014,” Osteopathic Family Physician 6, no. 6 (2014).
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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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Preface 7 and science, and pursues
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CHAPTER 1 An Invisible World Figure
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1.1 • What Our Ancestors Knew 11
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1.2 • A Systematic Approach 17 Th
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1.2 • A Systematic Approach 19 Fi
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1.2 • A Systematic Approach 21 Ta
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1.3 • Types of Microorganisms 23
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1 • Summary 31 SUMMARY 1.1 What O
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1 • Review Questions 33 17. The p
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CHAPTER 2 How We See the Invisible
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2.1 • The Properties of Light 37
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2.1 • The Properties of Light 39
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2.2 • Peering Into the Invisible
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2.3 • Instruments of Microscopy 4
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2.4 • Staining Microscopic Specim
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2 • Review Questions 71 REVIEW QU
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CHAPTER 3 The Cell Figure 3.1 Micro
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3.1 • Spontaneous Generation 75 F
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3.2 • Foundations of Modern Cell
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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 • 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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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 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 • 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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