DNA Replication, Transcription, and Translation

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Questions and Answers

What is the function of helicase in DNA replication?

  • Unzips DNA (correct)
  • Seals gaps in DNA
  • Adds mRNA
  • Builds DNA strands

What enzyme builds DNA strands during replication?

  • DNA polymerase (correct)
  • Ligase
  • RNA polymerase
  • Helicase

What is the role of ligase in DNA replication?

  • Adding primers to DNA
  • Sealing gaps in DNA (correct)
  • Building DNA strands
  • Unzipping DNA

What does it mean for DNA replication to be 'semi-conservative'?

<p>Each new DNA molecule contains one original and one new strand. (D)</p>
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Which enzyme is responsible for making mRNA from DNA during transcription?

<p>RNA polymerase (B)</p>
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What cellular structure reads mRNA to assemble proteins?

<p>Ribosome (D)</p>
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Which molecule delivers amino acids to the ribosome for protein synthesis?

<p>tRNA (D)</p>
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What is the term for a change in the nucleotide sequence of DNA?

<p>Mutation (D)</p>
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What type of mutation does NOT change the amino acid sequence?

<p>Silent (B)</p>
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Which of the following mutations results in a stop codon being introduced too early?

<p>Nonsense (D)</p>
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What kind of mutation involves a shift in the reading frame of a gene?

<p>Frameshift (B)</p>
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What is one potential cause of mutations?

<p>Radiation (C)</p>
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What does the Ames test primarily detect?

<p>Mutagenic potential of chemicals (D)</p>
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What is the term for the uptake of free DNA by bacteria?

<p>Transformation (D)</p>
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Which process involves the transfer of DNA via a virus?

<p>Transduction (A)</p>
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What is the transfer of DNA between bacteria via a pilus called?

<p>Conjugation (D)</p>
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What is the process that destroys all microbes?

<p>Sterilization (A)</p>
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What is the process that reduces pathogens on surfaces called?

<p>Disinfection (D)</p>
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What is the term for the process of making something safe for skin by reducing pathogens?

<p>Antisepsis (B)</p>
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What are the standard autoclave conditions?

<p>121°C, 15 psi, 15 minutes (D)</p>
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How does an autoclave work to sterilize?

<p>Using steam heat to denature proteins (D)</p>
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Which of the following is a common item treated in an autoclave?

<p>surgical tools (A)</p>
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What temperature and time are used in pasteurization?

<p>72°C for 15 seconds (B)</p>
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What is the main purpose of pasteurization?

<p>Kill pathogens and slow spoilage (B)</p>
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Which of the following is a halogen used as a disinfectant?

<p>Bleach (C)</p>
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What is the effective range of alcohol concentration for disinfection?

<p>60-70% (B)</p>
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How do Quats (Quaternary Ammonium Compounds) work?

<p>Disrupting membranes (B)</p>
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Which of the following is an advantage of using Quats?

<p>Non-corrosive (D)</p>
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What type of disinfectant is hydrogen peroxide?

<p>Peroxide (C)</p>
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What is the function of a 1 micron filter?

<p>Removes protozoa (C)</p>
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What does HEPA filter remove?

<p>Particles from air (D)</p>
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How does UV radiation kill bacteria?

<p>Damaging DNA (A)</p>
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What is a common effect of cold temperatures on microbes?

<p>Slowed metabolism (C)</p>
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What is the mechanism of action for penicillin?

<p>Cell wall inhibitor (C)</p>
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Which drug class inhibits protein synthesis?

<p>Tetracyclines (C)</p>
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How does ciprofloxacin work?

<p>Inhibits DNA replication (B)</p>
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What is the mechanism of action for sulfa drugs?

<p>Folic acid synthesis inhibition (A)</p>
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Enzymes that inactivate penicillin are called:

<p>Beta-lactamases (A)</p>
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What is a mechanism of antibiotic resistance where bacteria pump the drug out of the cell?

<p>Drug efflux (D)</p>
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What is the Kirby-Bauer test used for?

<p>Measuring zones of inhibition (D)</p>
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What does the zone of inhibition indicate in the Kirby-Bauer test?

<p>Antibiotic sensitivity (C)</p>
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Which two types of alcohol are commonly used as disinfectants?

<p>Isopropanol and ethanol (C)</p>
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What did Griffith's experiment demonstrate?

<p>Transformation in bacteria (A)</p>
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What did Avery's discovery determine?

<p>DNA is the genetic material (D)</p>
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What does the E-test determine?

<p>The minimum inhibitory concentration (MIC) of an antimicrobial drug (B)</p>
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How is the MIC determined in a broth dilution test?

<p>By observing the lowest concentration of drug that stops growth (C)</p>
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Which of the following is considered a sterile site in the human body?

<p>Blood (D)</p>
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Where is normal flora typically found in the body?

<p>Skin (C)</p>
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Which stage of disease occurs immediately after the incubation period?

<p>Prodromal (C)</p>
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What is a fomite?

<p>An inanimate object that transmits disease (C)</p>
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Which of the following is a possible reservoir for pathogens?

<p>Soil (A)</p>
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Which of the following is a common healthcare-associated infection (HAI)?

<p>Clostridium difficile infection (B)</p>
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Which characteristic describes exotoxins?

<p>Secreted and very toxic (D)</p>
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What part of Gram-negative bacteria is endotoxin?

<p>Lipopolysaccharide (LPS) (C)</p>
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What is the function of adhesins?

<p>To help bacteria attach to host cells (A)</p>
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How do capsules contribute to bacterial virulence?

<p>By evading the immune system (C)</p>
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What does antigenic variation allow a pathogen to do?

<p>Evade the immune system (A)</p>
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What is a gene?

<p>A segment of DNA that codes for a protein or RNA (B)</p>
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What is a plasmid?

<p>A small, circular piece of DNA carrying extra genes (B)</p>
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Which of the following describes a bacterial chromosome?

<p>Large and circular (C)</p>
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What does a bacteriophage infect?

<p>Bacteria (C)</p>
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What is the primary function of a cytokine?

<p>Immune signaling (C)</p>
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What does PAMP stand for?

<p>Pathogen-Associated Molecular Pattern (B)</p>
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What recognizes PAMPs?

<p>PRRs (B)</p>
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Which of these is an inflammatory cytokine?

<p>IL-1 (D)</p>
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What is opsonization?

<p>Tagging microbes for phagocytosis (D)</p>
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What is the function of the MAC?

<p>To lyse microbes (C)</p>
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What is the correct order of disease stages?

<p>Incubation → Prodromal → Illness → Decline → Convalescence (B)</p>
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What is the name for the measure of the lowest concentration of a drug that stops growth?

<p>Minimum Inhibitory Concentration (A)</p>
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What is Candida Auris?

<p>A drug-resistant yeast (A)</p>
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Exotoxins are secreted by which of the following?

<p>Bacteria (A)</p>
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What is the purpose of coagulase?

<p>Help bacteria evade the immune system (A)</p>
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What is kinase?

<p>A virulence enzyme (A)</p>
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CSF stands for:

<p>Cerebral Spinal Fluid (A)</p>
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Where is lipopolysaccharide found?

<p>Gram-negative bacteria (A)</p>
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Botulinum toxin is:

<p>A type of exotoxin (C)</p>
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What does it mean for transformation to occur?

<p>Bacteria uptakes genetic material (A)</p>
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What is the point where bacterial growth touches the strip called?

<p>Minimum inhibitory concentration (A)</p>
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Flashcards

Semi-conservative DNA replication

Each strand of DNA serves as a template for a new strand.

Helicase

Unzips DNA, separating strands.

DNA polymerase

Builds new DNA strands.

Ligase

Seals gaps in DNA strands.

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Transcription

RNA polymerase makes mRNA from DNA.

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Translation

Ribosomes read mRNA, and tRNA delivers amino acids to make proteins.

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Silent Mutation

No change in amino acid sequence.

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Missense Mutation

Results in a wrong amino acid.

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Nonsense Mutation

Results in a premature stop codon.

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Frameshift Mutation

Shifts the reading frame of the genetic code.

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Transformation (in bacteria)

Uptake of free DNA by bacteria.

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Transduction (in bacteria)

Virus transfers DNA between bacteria.

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Conjugation (in bacteria)

DNA transfer via plasmid through a pilus.

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Sterilization

Destroys all microbes.

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Disinfection

Reduces pathogens on surfaces.

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Antiseptics

Safe for use on the skin to reduce microbes.

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Autoclave

Denatures proteins via steam heat.

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Pasteurization

Kills pathogens & slows spoilage but doesn't sterilize.

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Halogens (as disinfectants)

Effective against bacteria, viruses, and fungi.

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Alcohols (as disinfectants)

Denatures proteins with water.

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Quats (as disinfectants)

Disrupt membranes.

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Peroxides (as disinfectants)

Forms reactive oxygen species (ROS).

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1 micron filter

Removes protozoa and most bacteria.

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How alcohols work

Denatures proteins and dissolves lipids.

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Radiation (UV)

Damages DNA via thymine dimers.

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Ionizing Radiation

Forms toxic oxygen which damages DNA.

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Cold

Slows metabolism and prevents reproduction.

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Penicillin

Penicillin inhibits cell wall synthesis.

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Drug resistance mechanism of efflux

Drug efflux pumps drug out of the cell.

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Broth dilution test for MIC

Lowest concentration to stop growth.

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Normal flora locations

Skin, GI tract, mouth, throat.

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Transmission routes

Airborne, foodborne, vector, fomite, direct contact.

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Descriptive Epidemiology

Description of disease patterns.

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Experimental Epidemiology

Test hypotheses about disease causes.

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Exotoxins

Secreted, very toxic proteins.

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Adhesins (fimbriae)

Surface structures for attachment.

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Swab Plate Evenly

Lab method to prep bacterial lawn.

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E-test

A lab method using a plastic strip with antibiotic gradient to find the Minimum Inhibitory Concentration (MIC).

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Sterile Sites

Blood, cerebrospinal fluid and lungs.

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Disease Stages

Incubation, Prodromal, Illness, Decline, and Convalescence.

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Reservoirs

Humans, animals, soil, and water.

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HAI Organisms

Clostridium difficile, Candida Auris, MRSA, E. coli, Pseudomonas.

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Gene

A segment of DNA that contains the instructions to make a specific protein or functional RNA.

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Plasmid

A small, circular piece of DNA found in many bacteria that exists separately from the chromosome and can carry extra genes.

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Chromosome

A large, organized structure of DNA that contains many genes.

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Bacteriophage

Virus that infects bacteria.

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Cytokine

Immune signaling molecule.

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PAMP

Pathogen signal recognized by host.

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PRR

Host receptor for PAMP.

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IL-1, TNF-alpha

Inflammatory cytokines.

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Opsonization

Tags microbes for phagocytosis.

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MAC

Membrane attack complex, lyses microbes.

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Clostridium difficile

Healthcare-associated infections caused by endospore-forming bacteria, leads to diarrhea.

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Candida Auris

A drug-resistant yeast causing healthcare-associated infections.

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Alcohols

Effective when diluted with water to denature proteins.

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Griffith’s Experiment

Dead virulent + live harmless bacteria = disease (proved transformation).

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Avery’s Discovery

DNA is the genetic material.

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Study Notes

DNA Replication

  • DNA replication is semi-conservative, meaning each strand serves as a template for a new strand
  • Key enzymes in DNA replication include:
    • Helicase: Unzips the DNA double helix
    • DNA polymerase: Builds the new DNA strands
    • Ligase: Seals gaps in the newly synthesized DNA

Transcription

  • RNA polymerase is the enzyme responsible for making mRNA from a DNA template

Translation

  • Ribosomes read mRNA sequences
  • tRNA molecules deliver corresponding amino acids to build a protein

Prokaryotes vs. Eukaryotes

  • Prokaryotes
    • Lack a nucleus
    • Transcription and translation occur simultaneously
  • Eukaryotes
    • Transcription occurs in the nucleus
    • Translation occurs in the cytoplasm

Types of Mutations

  • Silent mutation: No change in the amino acid sequence
  • Missense mutation: Results in a wrong amino acid being incorporated
  • Nonsense mutation: Introduces a stop codon prematurely, truncating the protein
  • Frameshift mutation: Shifts the reading frame, altering the entire amino acid sequence downstream

Causes of Mutations

  • Radiation
  • Chemicals
  • Copying errors during DNA replication

Ames Test

  • Used to detect the mutagenic potential of chemicals

DNA Sharing in Bacteria

  • Transformation: Uptake of free DNA from the environment
  • Transduction: Transfer of DNA via a virus
  • Conjugation: Transfer of DNA via a plasmid through a pilus

Griffith's Experiment

  • Demonstrated transformation
    • Dead virulent bacteria + live harmless bacteria = disease (proved transformation)

Avery's Discovery

  • Identified DNA as the genetic material

Microbial Control - Definitions

  • Sterilization: Destroys all microbes
  • Disinfection: Reduces pathogens on surfaces
  • Antiseptics: Safe for use on skin

Autoclave Conditions

  • 121°C, 15 psi, for 15 minutes
  • Denatures proteins via steam heat
  • Used for liquids, glassware, surgical tools, and some plastics
  • Time can vary from 20-45 minutes

Pasteurization

  • 72°C for 15 seconds (does not boil)
  • Kills pathogens, slows spoilage, but doesn't stop it completely

Disinfectant Categories & Examples

  • Halogens
    • Bleach
    • Effective against bacteria, viruses, and fungi
    • Do not kill all endospores or cysts
  • Alcohols
    • Isopropanol
    • Ethanol
    • Effective range: 60-70% (with water for protein denaturation)
  • Quats (Quaternary Ammonium Compounds)
    • Benzalkonium chloride
    • Disrupt membranes
    • Non-corrosive, non-staining, no strong fumes, inexpensive, and easy to use
    • Some microbes (e.g., Pseudomonas) can grow in quats
  • Peroxides
    • Hydrogen peroxide
  • Filters
    • 1 micron: Removes protozoa and most bacteria
    • HEPA: Filters air in rooms
    • N95: Filters air for individual breathing

How Disinfectants Work

  • Alcohols: Denature proteins and dissolve lipids (effective against envelope viruses)
  • Autoclave: Steam denatures proteins

Radiation (UV)

  • Damages DNA via thymine dimers

Ionizing Radiation

  • Forms toxic oxygen via ionized water and damages DNA

Cold

  • Slows metabolism and prevents reproduction

Filtration

  • Physically removes microbes from liquid or air

Antimicrobial Drugs - Common Drug Classes

  • Penicillin: Cell wall inhibitor (e.g., for strep infections)
  • Tetracycline: Protein synthesis inhibitor (e.g., for acne)
  • Ciprofloxacin: DNA replication inhibitor (e.g., for UTIs)
  • Sulfa drugs: Folic acid synthesis inhibitor

Resistance Mechanisms

  • Drug inactivation: Beta-lactamase inactivates penicillin
  • Drug efflux: Pumps drug out of the cell (e.g., tetracycline resistance)
  • Blocked entry: Modified porins in Gram-negative bacteria
  • Target alteration: Changes in ribosomes (e.g., tetracycline, aminoglycoside resistance)

Testing Methods – Kirby-Bauer

  • Measures the zone of inhibition around antibiotic disks

E-test (Epsilometer test)

  • Determines the Minimum Inhibitory Concentration (MIC)
    • A plastic strip with a gradient of antibiotic concentrations is placed on an agar plate with bacteria
    • As the bacteria grow, an elliptical zone of inhibition forms around the strip.
    • The point where bacterial growth touches the strip indicates the MIC—the lowest concentration of the drug that inhibits visible bacterial growth

Broth Dilution

  • Determines the lowest concentration to stop growth (MIC)
    • Bacteria exposed to serial dilutions of drug in 96-well plate
    • MIC is the lowest concentration with no visible growth

Antimicrobial Drug Examples

  • Penicillin: Inhibits cell wall synthesis, used for streptococcal infections; resistance via beta-lactamase
  • Tetracycline: Inhibits protein synthesis, used for acne and respiratory infections; resistance via efflux pumps
  • Ciprofloxacin: Inhibits DNA replication, used for UTIs and anthrax; resistance via altered target sites
  • Sulfa drugs: Inhibits folic acid synthesis, used for UTIs and respiratory infections; resistance via altered target sites
  • Common sensitivity tests include Kirby-Bauer, E-test, and Broth dilution

Epidemiology - Normal Flora Locations

  • Skin
  • GI tract
  • Mouth
  • Throat

Sterile Sites in Body

  • Blood
  • CSF
  • Lungs

Disease Stages

  • Incubation → Prodromal → Illness → Decline → Convalescence

Transmission Routes

  • Airborne
  • Foodborne
  • Vector
  • Fomite
  • Direct contact

Reservoirs

  • Human
  • Animal
  • Soil
  • Water

HAI (Healthcare-Associated Infection) Organisms & Diseases

  • Clostridium difficile: Endospores, causes diarrhea
  • Candida Auris: Drug-resistant yeast
  • MRSA, E. coli, Pseudomonas: pneumonia, wounds, UTIs

Key Organisms and Characteristics

  • Clostridium difficile
    • Gram-positive bacterium
    • Forms endospores, anaerobic
    • Causes severe diarrhea, colitis
  • Candida auris
    • Fungal yeast
    • Drug-resistant, invasive
    • Causes bloodstream, wound, and ear infections
  • MRSA (Staph. aureus)
    • Gram-positive bacterium
    • Methicillin-resistant
    • Causes pneumonia and wound infections
  • Escherichia coli
    • Gram-negative bacterium
    • Normal flora, opportunistic
    • Causes UTIs, pneumonia, and wound infections
  • Pseudomonas aeruginosa
    • Gram-negative bacterium
    • Resistant and forms biofilms
    • Causes pneumonia (esp. in CF), UTIs, and wounds

Descriptive vs. Experimental Epidemiology

  • Descriptive Epidemiology
    • Goal:Describe disease patterns (who, where, when)
    • Design: Observational
    • Questions: Who is affected? When and where?
    • Tools: Case reports, surveillance, epidemic curves
    • Example: Tracking measles outbreak in a school
    • Data Collection: Often retrospective or real-time
    • Use: Generate hypotheses
  • Experimental Epidemiology
    • Goal: Test hypotheses about causes or treatments
    • Design: Controlled experiment
    • Questions: Does X cause Y? Does a treatment work?
    • Tools: Clinical trials, randomized controlled trials
    • Example: Testing a new vaccine in two randomized groups
    • Data Collection: Prospective (follows subjects forward in time)
    • Use: Confirm or refute hypotheses

Virulence & Pathogenicity - Toxins

  • Exotoxins: Secreted, very toxic (e.g., botulinum toxin)
  • Endotoxins: LPS of Gram-negatives, weakly toxic

Virulence Factors

  • Adhesins (fimbriae): Attachment
  • Capsules: Evade immune system
  • Enzymes (coagulase, kinase): Damage tissues
  • Antigenic variation (e.g., HIV, Trypanosoma)
  • Invasion into host cells

Virulence Factors - Examples

  • Adhesins (fimbriae)
    • Surface structures for attachment
    • E. coli, Neisseria gonorrhoeae
    • Helps microbes stick to host tissues
  • Capsules
    • Polysaccharide coating that hides the cell
    • Streptococcus pneumoniae
    • Prevents phagocytosis by immune cells
  • Enzymes
    • Coagulase (clots), kinase (spreads infection)
    • Staphylococcus aureus
    • Disrupts tissues or evades immune response
  • Antigenic variation
    • Alters surface proteins to avoid detection
    • HIV, Trypanosoma
    • Evades immune memory and prolongs infection
  • Invasion
    • Forces entry into host cells
    • Salmonella, Listeria
    • Hides inside host cells, avoids immune attack

Key Vocabulary

  • Gene: Segment of DNA with instructions to make a specific protein or functional RNA
  • Plasmid: Small, circular piece of DNA in bacteria, separate from the chromosome, carries extra genes like antibiotic resistance
  • Chromosome: Large, organized structure of DNA containing many genes
  • Bacteriophage: Virus that infects bacteria
  • Cytokine: Immune signaling molecule
  • PAMP (Pathogen-Associated Molecular Pattern): Pathogen signal recognized by host
  • PRR (Pattern Recognition Receptor): Host receptor for PAMP
  • IL-1, TNF-alpha: Inflammatory cytokines
  • Opsonization: Tags microbes for phagocytosis
  • MAC: Membrane attack complex, lyses microbes

Kirby-Bauer Disk Diffusion Test Steps

  • Prepare a bacterial lawn
    • Use a sterile swab to evenly spread the test bacterium over the entire surface of a Mueller-Hinton agar plate
  • Let the plate dry
    • Allow the plate to sit for about 5–10 minutes so moisture absorbs
  • Place antibiotic-impregnated disks
    • Use sterile forceps or a disk dispenser, place antibiotic disks evenly spaced on the agar surface
    • Gently press each disk to ensure full contact
  • Invert and incubate the plate
    • Turn the plate upside down and incubate at 35–37°C for 16–18 hours
  • Measure zones of inhibition
    • After incubation, use a ruler or caliper to measure the diameter (in mm) of the clear zones around each disk
  • Interpret results
    • Compare measurements to a standard chart to determine if the bacterium is: Susceptible (S), Intermediate (I), or Resistant (R) to the antibiotic

Spectrophotometer

  • Measures turbidity to estimate bacterial growth

Disinfectant Lab

  • Halogens
    • Example: bleach, iodine
    • Mode of Action: oxidizes cell components
    • Effectiveness: Very effective
  • Phenolics
    • Example: Lysol
    • Mode of Action: disrupts membranes, denatures proteins
    • Effectiveness: Moderately effective
  • Alcohols
    • Example: Ethanol, isopropanol
    • Mode of Action: Denatures proteins, dissolves membranes
    • Effectiveness: Effective (requires water)
  • Quats (Quaternary ammonium)
    • Example: Benzalkonium chloride
    • Mode of Action: Disrupts membrane integrity
    • Effectiveness: Less effective against gram-negatives
  • Peroxides
    • Example: Hydrogen peroxide
    • Mode of Action: Forms reactive oxygen species (ROS)
    • Effectiveness: Highly effective (broad spectrum)
  • Heavy Metals
    • Example: Silver nitrate
    • Mode of Action: Inactivates proteins
    • Effectiveness: Effective but less commonly used

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