Chapter 7

Microbial Nutrition, Ecology, and Growth

Chapter Outline

7.1. Microbial Nutrition

A. Introduction
1. Nutrition
2. Nutrients
a. Essential
b. Macronutrients
c. Micronutrients or “trace elements”
B. Chemical analysis of microbial cytoplasm
1. Water
2. Proteins
3. Organic compounds
4. Chemical elements
C. Sources of essential nutrients
1. Carbon sources
a. Heterotroph
b. Autotroph
2. Nitrogen sources
3. Oxygen sources
4. Hydrogen sources
a. Maintain pH
b. Form hydrogen bonds between molecules
c. Serve as source of free energy in oxidation-reduction reactions
5. Phosphorus (phosphate) sources
6. Sulfur sources
7. Other nutrients important in microbial metabolism
8. Growth factors: Essential organic nutrients
a. Include amino acids, a nitrogenous base, or a vitamin that cannot be
synthesized by an organism.
b. Haemophilus influenzae
9. How microbes feed: Nutritional types
a. Phototrophs
b. Chemotrophs
10. Autotrophs and their energy sources
a. Photoautotrophs
i. Plants, algae, some bacteria
b. Chemoautotrophs
i. Methanogens
c. Lithoautotrophs
11. Heterotrophs and their energy sources
a. Chemoheterotrophs
i. Aerobic respiration
ii. Fermentation
c. Saprobic microorganisms
i. Obligate saprobe
ii. Facultative parasite

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 iii. Opportunistic pathogens
d. Parasitic microorganisms
i. Pathogens
ii. Obligate parasites
iii. Obligate intracellular parasite
D. Transport mechanisms for nutrient absorption
E. The movement of water: Osmosis
1. Selectively or differentially permeable membranes
2. Tonicity conditions (concentration)
a. Isotonic
b. Hypotonic
c. Hypertonic
d. Direction of net water movement and osmotic pressure
3. Adaptations to osmotic variations in the environment
F. The movement of molecules: Diffusion and transport
1. Diffusion
a. Simple diffusion
b. Facilitated diffusion
c. Specificity
d. Saturation
2. Active transport: Bringing in molecules against a gradient
a. Transport of nutrients against the diffusion gradient
b. Transport of nutrients in the same direction as the natural gradient but faster
than diffusion alone.
c. Presence of specific membrane proteins
d. Expenditure of energy
e. Examples:
i. Sodium, potassium, and hydrogen pumps
ii. Group translocations
G. Endocytosis: Eating and drinking by cells
a. Phagocytosis
b. Pinocytosis
7.2. Environmental Factors That Influence Microbes
A. Temperature adaptations
1. Minimum, optimum, and maximum temperature
2. Psychrophile
3. Mesophile
4. Thermophile
B. Gas requirements
1. How microbes process oxygen
a. Aerobe
b. Obligate aerobe
c. Facultative anaerobe
d. Microaerophile
e. Anaerobe
f. Strict or obligate anaerobe
g. Aerotolerant anaerobes
2. Capnophiles—higher CO2
C. Effects of pH
1. Acidophiles
2. Alkalinophiles

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 D. Osmotic pressure
1. Halophiles
2. Facultative halophiles
E. Miscellaneous environmental factors
1. Barophiles
2. Dehydrated cell stages
F. Ecological associations among microorganisms
1. Symbiosis and symbionts
2. Mutualism
3. Commensalism and satellitism
4. Parasitism
5. Synergism
6. Antagonism
a. Antibiosis
G. Interrelationships between microbes and humans
1. Normal microbial flora
2. Lactobacillus spp. in vagina
7.3. The Study of Microbial Growth
A. The basis of population growth: Binary fission
B. The rate of population growth
1. Generation or doubling time
2. Mycobacterium leprae and Salmonella enteriditis
3. Exponential: Geometric increase
C. The population growth curve
D. Stages in the normal growth curve (Fig. 7.15)
1. Lag phase
2. Exponential or logarithmic (log) phase
3. Stationary phase
4. Death phase
5. Practical importance of the growth curve
i. Chemostat
E. Other methods of analyzing population growth
1. Turbidity assessment
2. Enumeration of bacteria
a. Direct or total cell count
b. Cytometer counting
c. Coulter counter

Key Terms and Phrases

Nutrition Chemotroph Passive transport

Nutrients Photoautotroph Active transport
Essential nutrient Chemoautotrophs Bulk transport
Macronutrients Lithoautotrophs Diffusion
Micronutrients Chemoheterotrophs Osmosis
Trace elements Saprobes Permeable
Inorganic nutrient Parasites Isotonic
Organic nutrient Obligate Hypotonic
Heterotroph Facultative Hypertonic
Autotroph Opportunistic Facilitated diffusion
Phototroph Pathogens Group translocation

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Endocytosis Aerotolerant Normal microbial flora
Phagocytosis Capnophiles Binary fission
Pinocytosis Acidophiles Generation or doubling time
Minimum temperature Alkalinophiles Exponential
Maximum temperature Halophiles Lag phase
Optimum temperature Barophiles Exponential (log) phase
Psychrophile Symbiosis Stationary phase
Mesophile Mutualism Death phase
Thermophile Commensalism Turbid
Aerobe Parasitism Chemostat
Anaerobe Synergism Direct (total) cell count
Microaerophile Antagonism

Topics for Discussion

The most complex issue for most students, in this chapter, continues to be the concept of osmotic
pressure. The terms isotonic, hypotonic, and hypertonic should be reviewed. The students need a clear
understanding of these terms to apply the material in this chapter. Additionally, students will need
reinforcing of nutritional requirement concepts (autotrophs, heterotrophs, etc.). Students could be placed
in discussion groups and directed to create lists of specific organisms and how they would best grow
them. The clinical infective applications of the growth curve and the rate of infection spread in humans
should be discussed.

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 Chapter 8
Microbial Metabolism: The Chemical Crossroads of Life

Chapter Outline

8.1. The Metabolism of Microbes

A. Metabolism
1. Anabolism
2. Catabolism
3. Metabolism:
a. Assembles smaller molecules into larger macromolecules for the cell
b. Degrades macromolecules into smaller molecules and yields energy
c. Conserves energy in the form of ATP or heat
B. Enzymes: Catalyzing the chemical reactions of life
1. How do enzymes work?
a. Catalysts
b. Energy of activation
c. Substrates
2. Enzyme structure
a. Simple enzymes
b. Conjugated enzymes
c. Holoenzyme
i. Apoenzyme
ii. Cofactors
1. Coenzymes (organic molecules)
2. Inorganic elements (metal ions)
3. Apoenzymes: Specificity and the active site
a. Active site: where the substrate binds
b. Crevice or groove on the enzyme
4. Enzyme-substrate interactions
a. Temporary union between enzyme and substrate
b. Weak, reversible bonding
c. Reaction occurs on substrate
d. Product is formed and released
5. Cofactors: Supporting the work of enzymes
a. Metallic cofactors
b. Coenzymes and vitamins
6. Classification of enzyme functions
a. Location and regularity of enzyme action
i. Exoenzymes
ii. Endoenzymes
iii. Constitutive enzymes
iv. Regulated enzymes
b. Synthesis and hydrolysis reactions
i. Dehydration reactions
ii. Hydrolysis reaction
c. Transfer reactions by enzymes
i. Oxidation and reduction
ii. Aminotransferases
iii. Phosphotransferases
iv. Methyltransferases

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 v. Decarboxylases
7. The role of microbial enzymes in disease
a. Streptokinase: Streptococcus pyogenes
b. Elastase and collagenase: Pseudomonas aeruginosa
c. Lecithinase C: Clostridium perfringens
8. The sensitivity of enzymes to their environments
a. Labile
b. Denaturation
C. Regulation of enzymatic activity and metabolic pathways
1. Metabolic pathways
a. Multistep series of reactions
b. Interconnected and merge at many sites
2. Direct controls on the actions of enzymes
a. Competitive inhibition
b. Noncompetitive inhibition through negative feedback
3. Controls on enzyme synthesis
a. Enzyme repression
b. Enzyme induction: Escherichia coli
8.2. The Pursuit and Utilization of Energy
A. Energy in cells
1. Exergonic
2. Endergonic
B. A closer look at biological oxidation and reduction
1. Redox reactions
a. Redox pair: electron donor and electron acceptor
2. Phosphorylation and ATP synthesis
3. Electron transfer
4. Dehydrogenation
5. Electron carriers: Molecular shuttles
a. NAD and NADH
b. Final electron acceptor in aerobic metabolism is oxygen
c. In anaerobic metabolism, final electron acceptor is some other organic or
inorganic compound.
C. Adenosine triphosphate: Metabolic money
1. The molecular structure of ATP
a. Phosphate, sugar (ribose), and a base (adenine)
2. The metabolic role of ATP
a. Substrate-level phosphorylation
b. Oxidative phosphorylation
c. Photophosphorylation
8.3. The Pathways
A. Introduction
1. Metabolism
a. Enzymes
b. Catabolize
c. Precursor molecules
d. Anabolize
e. Reducing power
f. Energy
B. Catabolism: Getting materials and energy
1. Glycolysis

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 2. Aerobic respiration
a. Glycolysis
b. Krebs cycle
c. Respiratory chain
3. Anaerobic respiration
a. Glycolysis
b. Final electron acceptor is not oxygen
C. Aerobic respiration
1. Glucose: The starting compound
2. Glycolysis: The starting lineup
i. An anaerobic step
ii. Pyruvic acid synthesis
iii. Steps in the glycolytic pathway
b. Pyruvic acid: A central metabolite
c. The Krebs cycle: A carbon and energy wheel
i. The processing of pyruvic acid
ii. Role of acetyl coenzyme A
iii. Role of NADH
iv. Steps in the Krebs cycle
3. The respiratory chain: Electron transport and oxidative phosphorylation
a. Elements of electron transport: The energy cascade
i. ATP synthase
ii. Oxidative phosphorylation
b. Formation of ATP and chemiosmosis
c. Potential yield of ATPs from oxidative phosphorylation
4. Summary of aerobic respiration
a. Total possible yield
b. The terminal step
i. Formation of water
ii. Neisseria, Pseudomonas, Bacillus
iii. Klebsiella, Enterobacter
iv. Streptococcus
v. Superoxide dismutase and catalase
D. Anaerobic respiration
1. Nitrate and nitrite reduction systems
a. E. coli
2. Denitrification
a. Pseudomonas and Bacillus
E. Fermentation
1. Products of fermentation in microorganisms
a. Alcoholic fermentation
i. Converting pyruvic acid to ethanol
b. Acidic fermentation
i. Streptococcus and Lactobacillus: Homolactic fermentation
ii. Leuconostoc: Heterolactic fermentation
c. Mixed acid fermentation
i. Propionibacterium
F. Catabolism of noncarbohydrate compounds
1. Lipid catabolism:
a. Lipases: break down lipids to fatty acids and glycerol
b. Fatty acids are broken down through beta oxidation

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 2. Protein catabolism:
a. Proteases
b. Deamination
8.4. Biosynthesis and the Crossing Pathways of Metabolism
A. The frugality of the cell: Waste not, want not
1. Amphibolic sources of cellular building blocks
a. Gluconeogenesis
b. Beta-oxidation
c. Amination
d. Transamination
B. Anabolism: Formation of macromolecules
1. Carbohydrate biosynthesis
2. Amino acids, protein synthesis, and nucleic acid synthesis
C. Assembly of the cell
8.5 It All Starts with the Sun
A. Photosynthesis
1. Two phases
a. Light-dependent reactions require sunlight
b. Light-independent reactions proceed in light or dark conditions
c. Photons are energy packets from the sun
d. Photosynthetic pigments absorb energy:
i. Chlorophylls
ii. Carotenoids
iii. Phycobilins
2. Light-dependent reactions
a. Take place in the thylakoid membranes (grana) of the chloroplast
b. Photosystem I
c. Photosystem II
i. Photolysis
ii. Cytochromes
3. Light-independent reactions
a. RuBP
b. Carbon fixation
4. Other mechanisms of photosynthesis
a. Oxygenic
b. Anoxygenic

Key Terms and Phrases

Anabolism Coenzymes Regulatory

Catabolism Hydrolysis Enzyme repression
Metabolites Exoenzyme Enzyme induction
Enzymes Endoenzyme Energy
Catalysts Constitutive enzymes Endergonic
Energy of activation Induced enzymes Exergonic
Substrate Oxidized Redox reactions
Holoenzyme Reduced Phosphorylate
Apoenzyme Labile Dehydrogenation
Cofactors Denaturation Final electron acceptor
Active site Competitive inhibition Labile
Metallic cofactors Negative feedback NAD

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FAD ETS Transamination
ATP Fermentation Beta oxidation
Glycolysis Amphibolism Chemiosmosis
Krebs Cycle Photosynthesis Gluconeogenesis
Aerobic respiration Light-dependent reactions Photolysis
Ferment Light-independent reactions RuBP
Anaerobic respiration Chlorophylls Carbon fixation
Pyruvic acid Amination Chloroplast

Topics for Discussion

A review of cellular metabolism, both anabolic and catabolic, will help the student with the material in
this chapter. Students should have had some coverage of this material from prerequisite courses.
Students should compare the effectiveness of aerobic versus anaerobic reactions that produce energy.
Discuss the role of vitamins, minerals, and nucleotides in energy production. Create a list of useful
products of fermentation. The process of photosynthesis can be discussed in terms of its importance to
the development of life on earth.

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 Chapter 9
Microbial Genetics

Chapter Outline

9.1. Introduction to Genetics and Genes: Unlocking the Secrets of Heredity

A. Genetics is the study of the heredity of living things
1. Includes
a. Transmission of traits
b. Expression and variation of those traits
c. Structure and function of the genetic material
d. How this material changes
2. The nature of the genetic material
3. Levels of structure and function of the genome
a. Genome
b. Chromosome
c. Gene
i. DNA that codes for a protein or RNA molecule
ii. Structural genes
iii. RNA genes
iv. Regulatory genes
d. Genotype and phenotype
4. Size and packaging of genomes
B. The DNA code: A simple yet profound message
1. Nucleotide
a. Consists of a phosphate group, deoxyribose sugar and a nitrogenous base
2. Purines and pyrimidines: Adenine, guanine, cytosine, and thymine
3. Antiparallel arrangement in DNA
C. The significance of DNA structure
1. Maintaining the code
2. Providing variety
D. DNA replication: Preserving the code and passing it on
1. Overall replication process
a. Templates and enzymes
b. Semiconservative replication
2. Refinements and details of replication
a. Helicases
b. DNA polymerase
c. RNA primer
d. Replication fork
e. Leading strand
f. Lagging strand
g. Okazaki fragments and DNA ligase
3. Elongation and termination of the daughter molecules
4. Replication in other biological systems
9.2. Applications of the DNA Code: Transcription and Translation
A. Introduction
1. Transcription: RNA copy of DNA sequence
2. Translation: information in RNA is used to make protein
3. “Central dogma”

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 B. The gene-protein connection
1. The triplet code and the relationship to proteins
a. Structural genes
b. Phenotype
C. The major participants in transcription and translation
1. RNAs: Tools in the cell's assembly line
a. Single-stranded molecule
b. Composed of nucleotides with a phosphate group, ribose sugar, and a
nitrogenous base (uracil, adenine, guanine, and cytosine)
c. Include mRNA, tRNA, rRNA, miRNA (micro), riboswitch RNA, and siRNA
(small interfering)
2. Messenger RNA: Carrying DNA's message
a. Codons: a sequence of three nucleotides
3. Transfer RNA: The key to translation
a. Anticodon: complements mRNA codons
4. The ribosome: A mobile molecular factory for translation
a. Prokaryotic ribosome (70S)
D. Transcription: The first stage of gene expression
1. RNA polymerase
2. Template strand
3. Coding strand
4. Promoter region
5. mRNA transcript
E. Translation: The second stage of gene expression
1. The initiation of translation
2. The master genetic code: The message in messenger RNA
a. Redundancy and “wobble”
3. The beginning of protein synthesis
a. Start codon: AUG
4. Continuation and completion of protein synthesis: Elongation and termination
a. tRNA
b. Translocation
c. Termination (nonsense) codons
d. Posttranslational modifications
e. Polyribosomal complex
F. Eucaryotic transcription and translation: Similar yet different
1. Introns
2. Exons
3. Split gene
4. Spliceosome
G. The genetics of animal viruses
9.3. Genetic Regulation of Protein Synthesis and Metabolism
A. The lactose operon: A model for inducible gene regulation in bacteria
1. Regulator
a. Repressor
b. Allosteric
2. Control locus
a. Promoter
b. Operator
3. Structural locus
4. Inducer

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 B. A repressible operon
1. Corepressor
C. Antibiotics that affect transcription and translation
1. Actinomycin D
2. Erythromycin and spectinomycin
3. Chloramphenicol
4. Streptomycin
9.4. Mutations: Changes in the Genetic Code
A. Introduction
1. Mutation
2. Wild type strain
3. Mutant strain
B. Causes of mutations
1. Spontaneous mutation
2. Induced mutation
a. Mutagens
C. Categories of mutations
1. Point mutations
2. Missense mutation
3. Nonsense mutation
4. Silent mutation
5. Back-mutation
6. Frameshift
D. Repair of mutations
1. Photoactivation or light repair
2. Excision repair
a. Xeroderma pigmentosa and the enzyme photolyase
3. Mismatched bases
E. The Ames Test: Salmonella typhimurium
F. Positive and negative effects of mutations
1. Adaptation
2. Natural selection
9.5. DNA Recombination Events
A. Transmission of genetic material in bacteria
1. Conjugation: Bacterial “sex”
a. Fertility or F factor
b. Conjugative pilus
c. Donor and recipient
d. Resistance plasmids or R factors
2. Transformation: Capturing DNA from solution
a. Streptococcus pneumoniae
b. Transformation
c. Competent
d. Transfection
3. Transduction: The case of the piggyback DNA
a. Generalized transduction
b. Specialized transduction
c. Corynebacterium diphtheriae
4. Transposons: "This gene is jumpin''

Key Terms and Phrases

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Genetics tRNA Exons
Heredity rRNA Split gene
Genome Genotype DNA polymerase
DNA Phenotype Corepressor
RNA Single stranded Mutation
Chromosome Double stranded Spontaneous mutation
Deoxyribose sugar Codons Missense mutation
Nitrogenous base Anticodon Point mutation
Purine RNA polymerase Induced mutation
Pyrimidine Promoter region Nonsense mutation
Adenine Start codon Photoactivation
Thymine Translocation Ames Test
Guanine Nonsense codons Conjugation
Cytosine Triplets Transformation
Uracil Carcinogenic Transduction
Replication Operon Sex pilus
Okazaki fragments Locus Transposons
Transcription Posttranslational Allosteric
Translation modifications
mRNA Introns

Topics for Discussion

The students will appreciate a review of the nature of the genetic material. A good historical discussion
can center on the work of Watson, Crick, Wilkins, and Rosalind Franklin. Animations, 3D graphics, and
interactive tutorials are available on the Internet and will aid in the students' understanding of the
replication process. A good discussion of mutations and adaptation will include the topic of antimicrobial
resistance in bacteria and its increase in hospital as well as community settings.

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 Chapter 10
Genetic Engineering: A Revolution in Molecular Biology

Chapter Outline

10.1. Basic Elements and Applications of Genetic Engineering

A. Genetics is considered basic science
B. Genetic engineering is an applied science
10.2. Tools and Techniques of Genetic Engineering
A. DNA: The raw material
1. Enzymes for dicing, splicing, and reversing nucleic acids
a. Restriction endonucleases
i. Palindromes
ii. EcoRI from E. coli and HindIII from Haemophilus influenzae
iii. Restriction fragments and restriction fragment length
polymorphisms (RFLPS)
b. Ligase
c. Reverse transcriptase
d. Complementary DNA (cDNA)
2. Analysis of DNA
a. Gel electrophoresis
3. Nucleic acid hybridization and probes
a. Gene probes
b. Southern blot method
c. FISH
4. Methods used to size, synthesize, and sequence DNA
a. DNA sequencing: Determining the exact genetic code
i. Sanger method
5. Polymerase chain reaction: A molecular Xerox machine for DNA
a. Primers
b. DNA polymerases
i. Thermus aquaticus and Thermococcus litoralis
c. Steps in PCR cycle:
i. Denaturation
ii. Priming
iii. Extension
10.3. Methods in Recombination DNA Technology: How to Imitate Nature-Or to “One-Up” It
A. Introduction
1. Recombinant DNA technology
a. Clones
b. Vectors
c. Cloning hosts
B. Technical aspects of recombinant DNA and gene cloning
1. Characteristics of cloning vectors
a. Charon phage
b. Cosmids
c. Artificial chromosomes
2. Characteristics of cloning hosts
a. E. coli
b. Saccharomyces cerevesiae

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 c. Bacillus subtilis
C. Construction of a recombinant, insertion into a cloning host, and genetic expression
10.4. Biochemical Products of Recombinant DNA Technology
A. Human growth hormone
B. Insulin
C. Clotting factor VIII
10.5. Genetically Modified Organisms
A. Recombinant microbes: Modified bacteria and viruses
1. Pseudomonas syringae and Frostban
2. Pseudomonas fluorescens, Bacillus thuringiensis and insecticide genes
B. Transgenic plants: Improving crops and foods
1. Agrobacterium
C. Transgenic animals: Engineering embryos
1. Pharming
10.6. Genetic Treatments: Introducing DNA into the Body
A. Gene therapy
1. Adenosine deaminase (ADA) deficiency
2. SCID
B. DNA technology as genetic medicine
1. Antisense DNA: Targeting messenger RNA
10.7. Genome Analysis: Maps, Fingerprints, and Family Trees
A. Genome mapping and screening: An atlas of the genome
1. Locus and alleles
2. Linkage, physical and sequence maps
3. Genomics and bioinformatics
B. DNA fingerprinting: A unique picture of a genome
1. Single nucleotide polymorphism
2. Markers
a. Variable number of tandem repeats (VNTRs)
b. Microsatellite polymorphisms
3. Microarray analysis

Key Terms and Phrases

Genetic engineering Recombinant DNA Transgenic animals

Biotechnology technology Gene therapy
Germline therapy Clones Antisense RNA
Restriction endonuclease Vector Gene mapping
Ligase Cloning host DNA fingerprinting
Reverse transcriptase Yeast artificial chromosomes Forensic science
Complementary DNA Chimera Cleave
Gel electrophoresis Transformation Fluorescent in situ
Hybridize Transcription hybridization
Gene probes Translation Origin of replication
Polymerase chain reaction Pharming Bioterrorism
Primers "Superweeds" Bioethics
DNA polymerase Transfection Genomic libraries
Amplicons Plasmid

Topics for Discussion

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A major topic of discussion should be on the heightened awareness of bioterrorism and what impact
advances in biotechnology have had on this issue. Student discussions should include modern products,
including the Hepatitis B vaccine and why they should all receive it. Students should be encouraged to
list current uses of recombinant DNA in medicine, criminal investigations, and paternity suits, as well as
hypothesize about future developments. Discuss possible positive and negative aspects of using
genetically modified crops and/or animals.

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 Chapter 11
Physical and Chemical Control of Microbes

Chapter Outline

11.1. Controlling Microorganisms

A. General considerations in microbial control
1. Sterilization
2. Disinfection
3. Antisepsis
4. Decontamination
B. Relative resistance of microbial forms
1. Highest: prions, endospores
2. Moderate: Protozoan cysts, sexual fungal spores, naked viruses, Mycobacterium
3. Least: Most bacterial vegetative cells, ordinary fungal spores, enveloped viruses,
yeasts, and protozoan trophozoites
C. Terminology and methods of microbial control
1. Sterilization
a. Sterile
b. Sterilants
c. Disinfection
d. Sepsis
e. Asepsis
f. Antiseptics
g. A note about prions
2. The agents versus the process
a. –cide
i. Bactericide
ii. Germicide
iii. Fungicide
iv. Virucide
v. Microbicide
b. –static
i. Bacteristatic
ii. Microbistatic
3. Decontamination
a. Sanitization
b. Degermation
4. Practical concerns in microbial control
D. What is microbial death?
1. Factors that affect death rate
a. Numbers
b. Nature of microorganism
c. Temperature and pH
d. Concentration of agent
e. Mode of action of agent
f. Presence of solvents, organic matter, and inhibitors
E. How antimicrobial agents work: Their modes of action
1. The effects of agents on the cell wall

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 a. Cells deprived of functional cell wall lyse easily
2. How agents affect the cell membrane
a. Surfactants lower surface tension of cell membranes
3. Agents that affect protein and nucleic acid synthesis
a. Ribosome function
b. DNA and RNA function
4. Agents that alter protein function
a. Denaturation of proteins
11.2. Methods of Physical Control
A. Heat as an agent of microbial control
1. Mode of action and relative effectiveness of heat
a. Moist heat
b. Dry heat
2. Heat resistance and thermal death of spores and vegetative cells
3. Practical concerns in the use of heat: Thermal death measurements
a. Thermal death time
b. Thermal death point
c. Common methods of moist heat control
i. Steam under pressure
1. Autoclave
ii. Nonpressurized steam
1. Tyndallization (intermittent sterilization)
iii. Pasteurization: Disinfection of beverages
1. Thermoduric microbes
2. Sterile milk
iv. Boiling water: Disinfection
b. Dry heat: Hot air and incineration
i. Incineration: most rigorous heat treatment
ii. Hot-air oven
iii. Dry oven
B. The effects of cold and desiccation
1. Cold retards activities of microbes
2. Dessication
3. Lyophilization
C. Radiation as a microbial control agent
1. Modes of action of ionizing versus non-ionizing radiation
2. Ionizing radiation: Gamma rays, X rays, and cathode rays
a. Cold sterilization
b. Applications of ionizing radiation
3. Non-ionizing radiation: Ultraviolet rays
a. Pyrimidine dimers
b. Applications of ultraviolet radiation
D. Decontamination by filtration: Techniques for removing microbes
1. Pore diameters
2. Applications of filtration
11.3. Chemical Agents in Microbial Control
A. Choosing a microbicidal chemical
1. Levels of chemical decontamination
a. High
b. Intermediate
c. Low

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B. Factors that affect the germicidal activity of chemicals
1. Nature of microorganism
2. Nature of material being treated
3. Degree of contamination
4. Time of exposure
5. Strength and chemical action of germicide
C. Germicidal categories according to chemical group
1. The halogen antimicrobial chemicals
a. Chlorine and its compounds
b. Chlorine compounds in disinfection and antisepsis
i. Hypochlorites
ii. Chloramines
c. Iodine and its compounds
i. Free iodine
ii. Iodophors (alcohol and iodine complex)
d. Applications of iodine solutions
i. Iodine tincture
1. Surgical antiseptic
ii. Iodophore
1. Betadine
a. Surgical handscrub
2. Phenol and its derivatives
a. Phenolics
b. Aromatic carbon ring with added functional groups
3. Applications of phenolics
a. Triclosan
4. Chlorhexidine
a. Surgical hand scrub
b. Obstetric antiseptic
5. Alcohols as antimicrobial agents
a. Applications of alcohols
b. Ethanol, isopropyl alcohol
6. Hydrogen peroxide and related germicides
a. Applications of hydrogen peroxide
i. 3% hydrogen peroxide as an antiseptic
ii. Vaporized hydrogen peroxide
iii. Ozone used to disinfect air and water
7. Chemicals with surface action: Detergents
a. Surfactants
i. Anionic detergents (negatively charged): limited microbicidal power
ii. Cationic detergents (positively charged): much more effective
b. Applications of detergents and soaps
i. Quaternary ammonium compounds:
1. Benzalkonium chloride
2. Zephiran
8. Heavy metal compounds
a. Oligodynamic action
b. Drawbacks:
i. Toxic to humans
ii. Allergic reactions
iii. Neutralized by biological fluids and wastes

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 iv. Resistance can develop in treated microbes
c. Applications of heavy metals
i. Merthiolate
ii. Metaphen
iii. Silver nitrate
9. Aldehydes as germicides
a. Glutaraldehyde; formaldehyde
b. Applications of the aldehydes
10. Gaseous sterilants and disinfectants
a. Ethylene oxide
b. Chlorine dioxide
c. Applications of gases and aerosols
11. Dyes as antimicrobial agents
a. Crystal violet and malachite green
b. Acridine dyes
12. Acids and alkalis

Key Terms and Phrases

Sterile Microbistatic Tinctures

Germicide Denature Halogens
Disinfection Moist heat Oligodynamic action
Asepsis Dry heat Glutaraldehyde
Antisepsis Thermal death time Formaldehyde
Sanitation Thermal death point Ethylene oxide
Decontamination Autoclave Irradiation
Sterilization Pasteurization Chlorine dioxide
Fungicide Thermoduric Filtration
Bactericide Incineration Dessication
Virucide Dry oven Lyophilization
Sporicide Radiation Phenolics
Bacteristatic Ionizing radiation Pyrimidine dimer
Degermation Non-ionizing radiation
Surfactant Cold sterilization

Topics for Discussion

Discuss why it is not wise to kill all microorganisms in certain areas. Bring to their attention the issue of
the pervasive use of antimicrobial soaps, and possible drawbacks to its overuse.
What are the most effective ways of eliminating harmful contaminating agents, especially when
dealing with contaminants such as HIV, anthrax, and prions? The issue of bioterrorism and the spate of
misinformation (stocking up on duct tape and plastic) will also encourage vigorous discussion.
Students will be interested in a general discussion on food handling and preparation and the
sterilization of medical equipment and supplies. An introduction to wound care is a good way to
introduce the chemical compounds (and their relative efficacies) for disinfection or antisepsis.

11-4