Course: Microbiology

Type: Lecture (4)

Date: 23-2-2013
Notes:
Chapter 13

Viruses, Viroid, and Prions

Acellular Microbes
Organism: Virus
Definition:
Microscopic
Acellular agent
Composed of nucleic acid
Surrounded by a protein coat
General Characteristics of Viruses
Obligatory intracellular parasites
Contain DNA or RNA
No ribosomes (can express protein)
No ATP-generating mechanism
Contain a protein coat (for protection)
Some enclosed by an envelope
Some have spikes
Most infect only specific types of cells in one host
Host range specific host attachment sites and cellular factors
(specific marker for different type of cells)
Properties of viruses
1. Obligate intracellular parasites (host has its full range)
Host: bacteria, fungi, algae, plants and animals
2. Ubiquitous (all around appear)
Everywhere with major impact on development of biological life
3. Ultramicroscopic (Nano size)
From 20nm to 450nm (diameter)
4. Acellular
Compact and economical
5. Dependent
Cannot fulfill the characteristics of life
6. Inactive Macromolecules
Active only inside the host (once stay outside or without host, it cannot survive)
7. With basic structures only
Protein shell: Capsid and Nucleic acids
8. Nucleic acid
Only DNA or RNA but not both
9. Nucleic acids can only be double-stranded or single-strand
Double-stranded DNA or RNA; Single-stranded DNA or RNA
10. High specificity
Molecules on virus surface impart high specificity for attachment to host cells
(each is different)
11. Host mechanisms to multiply (Making use of Host multiplication cycle instead
of have its own)
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Control of host cells genetic material and regulating the synthesis and
assembly of new viruses
12. Lack enzymes
Deprived of enzymes for most metabolic processes
13. Lack machinery
Deprived of machinery for synthesizing proteins
(Refer to TB P.372 fig 13.1 Virus sizes)
Virion Structure
Nucleic acid
DNA or RNA
Capsid
Capsomeres
Envelope
Spikes
Refer to fig 13.3

13.6

Viral Antigens
Viral proteins: antigenic glycoproteins (different virus,
different proteins)
Hemagglutinin receptor binding and membrane fusion
Neuraminidase required for viral replication
Virus Particle Flowchart
Capsid
Covering
Virus
particle
Central
core

Envelope
(not foung un all
viruses)
Nucleic acid
molecule(s)
(DNA or
RNA)
Matrix proteins
Enzymes (not found in
all viruses)

(Refer to TB P. 379 fig 13.7 Inoculation of an embryonated egg)

Embryonated egg is a living tissue, it can make use the large mechanism of the living
tissue to multiply

Virus Identification
Cytopathic effects
Serological tests
Detect antibodies against viruses in a patient
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Use antibodies to identify viruses in neutralization tests, viral

hemagglutination, and Western blot
Nucleic acids
PFLPs
PCR

Multiplication of Animal Viruses

1. Attachment: viruses attach to hosts cell membrane
2. Penetration by endocytosis of function
3. Uncoating by viral or host enzymes
4. Biosynthesis: production of nucleic acid and proteins (done by the host cell)
5. Maturation: nucleic acid and capsid proteins assemble (expressed by the host
cell)
6. Release by budding (enveloped viruses) or rupture

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Oncogenic Viruses (Examples)

Oncogenic DNA viruses
Adenoviridae
Herpesviridae
Poxviridae
Papovaviridae
Hepadnaviridae

Oncogenic RNA viruses

Retroviridae
Viral RNA is
transcribed to DNA,
which can integrate
into host DNA
HTLV 1
Latent and Persistent Viral Infections
HTLV 2
Virus remains in asymptomatic host cell or long
Cold sores, shingles
Disease process occur over a long period; generally
Subacute sclerosing panencephalitis (measles

periods
is fatal
virus)

Prions
Proteinaceous infectious particle
Inherited and transmissible by ingestion, transplant
and surgical instruments
Spongiform encephalopathies (BSE)
Sheep scrapie
Fatal familial insomnia
Mad cow disease
Chapter 20
Antimicrobial Drugs
Antimicrobial Drugs
Chemotherapy: the use of drugs to treat a disease
Antimicrobial drugs (*)
Antibiotic
Broad spectrum
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Narrow spectrum
Superinfection
Selective toxicity: killing harmful microbes without damaging the host

Terminology of Chemotherapy
Chemotherapeut Any chemical used in the treatment, relief, or prophylaxis of a
ic Drug
disease
Prophylaxis
Use of a drug to prevent imminent infection of a person at risk
Antimicrobial
The use of chemotherapeutic drugs to control infection
Chemotherapy
Antimicrobials
All-inclusive term for any antimicrobial drug, regardless of its
origin
Antibiotics
Substances produced by the natural metabolic processes of
some microorganisms that can inhibit or destroy other
microorganisms
Semisynthetic
Drugs that are chemically modified in the laboratory after being
Drugs
isolated from natural sources
Narrow
Antimicrobials effective against a limited array of microbial
Spectrum
types. For example, a drug effective mainly on gram-positive
(Limited
bacteria
Spectrum)
Broad Spectrum Antimicrobials effective against a wide variety of microbial
(Extended
types. For example, a drug effective against both gram-positive
Spectrum)
and gram-negative bacteria
Principles of Antimicrobial Therapy
Goal of antimicrobial chemotherapy
Destroys the infective agent
Without harming the hosts cells
Drugs must be able to
Be easy to administer
Able to reach the infectious agent anywhere in the body
Be absolutely toxic to the infectious agent; absolutely nontoxic to the host
(selective)
Remain in the body as long as needed and be safety and easily broken down
and excreted

Characteristics of Ideal Antimicrobial Drug (Selectively toxic to microbe)

Nontoxic to host cells
Bacteriocidal rather than Bacteriostatic
Relatively soluble
Long potent action
Does not lead to antimicrobial resistance (depend on drugs molecule)
Complement or assist hosts defenses (prevent allergy) (depend on host)
Remain active in host tissues and body fluid
Readily delivered to site of infection
Reasonably priced
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Does not cause allergy or predisposing host to other infections

The Origins of Antimicrobial Drugs

Metabolic products of aerobic bacteria and fungi
Inhibit the growth of competing microbes in the same habitat
Derived from
Bacteria in the genera Streptomyces and Bacillus
Molds in the genera Penicillium and Cephalosporium
Considerations before Antimicrobial Therapy (Three factors)
Nature of the infection microorganism
Susceptibility (or sensitivity) of the microorganism to various drugs
The overall medical condition of the patient
(Refer to table 20.1 20.2 [P.562])
Fungi are difficult to treat because it is type of Eukaryotes. More technique is needed
to decide a selective
Check Your Understanding (20-3, 20-4)
The Action of Antimicrobial Drugs
Bactericidal
Kill microbes directly
Bacteriostatic
Prevent microbes from growing
(Refer to table 20.3 [P.564])
Cell wall = Peptidoglycan

Inhibitors of Cell Wall Synthesis

Penicillin
Natural penicillins
Semisynthetic penicillins
Extended-spectrum penicillins
Prevent cross-linking of the peptidoglycans
Refer to 4.13b-c Bacterial cell walls
Gram-positive cell wall
Thick peptidoglycan
Teichoic acids
Gram-negative cell wall
Thin peptidoglycan
Outer membrane
Periplasmic space
Refer to P.567 fig 20.6
(a) Natural penicillins (susceptible to penicillinases)
(b)Semisynthetic penicillins
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-lactam ring
Inhibit the cell wall synthesis (Interfere the biosynthesis of the cell wall)
Refer to fig 20.7 [P.568]

-Lactam Antibiotics
Penicillin
Penicillinases-resistant penicillins
Penicillins + -lactamase inhibitors
Carbapenems
Substitute a C for an S, add a double bond
Monobactam
Single ring
(Refer to fig 20.9)

Inhibitors of Cell Wall Synthesis

Polypeptide antibiotics
Bacitracin
Topical application
Against gram-positives
Vancomycin
Glycopeptide
Important last line against antibiotic-resistant S.aureus
Antimycobacterial antibiotics
Isoniazid (INH)
Inhibits mycolic acid synthesis
Mycobacterium tuberculosis
Ethambutol
Inhibits incorporation of mycolic acid
Secondary drug to avoid drug resistance
Check your understanding (20-6 20-8)
Inhibitors of Protein Synthesis
Chloramphenicol
Broad spectrum
Binds 50S subunit; inhibits people bond formation
Aminoglycosides
Streptomycin, neomycin, gentamicin
Broad spectrum
Change shape of 30S subunit
Tetracyclines
Broad spectrum
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Interfere with tRNA attachment

Glycylcyclines
MRSA and Acinetobacter baumanii
Bind 30S subunit; inhibit translation

[Refer to P.561 fig 20.2]

Antifungal Drugs: Inhibition of Ergosterol Synthesis
Targets the fungal membrane
Principal sterols of fungal membrane: ergosterol
v.s. Animal membranes: cholesterol
Azoles
Miconazole
Triazole
Allylamines
For azole resistant
Refer to fig 20-16 [P.576]

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{Inhibit antibiotic drug, affect DNA making process, block nucleotide replication}
Antiviral Drugs: Enzyme Inhibitors
Protease inhibitors
Indinavir: HIV
Integrase inhibitors
HIV
Antiviral Drugs: Entry Inhibitors
Entry inhibitors (prevent entering the host)
Amantadine: influenza
Fusion inhibitors (prevent fuse and perform packaging)
Zanamivir: influenza
Block CCR5; HIV
Antiviral Drugs: Interferons
Prevent spread of viruses to new cells
Alpha interferon: Viral hepatitis
Imiquimod
Promotes interferon production
Test to Guide Chemotherapy
MIC: Minimal Inhibitory Concentration
MBC: Minimal Bactericidal Concentration
Antibiogram (against the concentration that in order to estimate the optimal
dosage)
Identifying the Agent (Collect Specimens before any antimicrobial drug is given)
Body fluids, sputum, stool
Testing for Drug Susceptibility
Kirby-Bauer technique
Surface of an agar plate is spread with bacteria
Small discs containing a prepared amount of antibiotic are plate
Zone of inhibition surrounding the discs is measured and compared with a
standard for each drug
Antibiogram provide data for drug selection
This method is less effective for anaerobic, fastidious, or slow-grow bacteria
(Refer to fig 20.17)
Technique for Preparation and Interpretation of Disc Diffusion Test
Test for Drug Susceptibility
Tube dilution tests (volume is same comparable)
More sensitive and quantitative than the Kirby-Bauer test
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Antimicrobial is diluted serially

Uniform sample of pure culture
Minimum inhibitory concentration (MIC): the smallest concentration
(highest dilution) of drug that visibly inhibits growth
A comparative index against other antimicrobials
Determine effective dosage

Tube Dilution Test for Determining MIC (Same inoculum size of test bacteria added)
Refer to fig 20.18, 20.19
Check Your Understanding 20-16
Resistance to Antimicrobial Drugs

(Refer to P.582 fig 20.21

The development of an antibioticresistant mutant during antibiotic
therapy)

Antibiotic Resistance
A variety of mutations can lead to antibiotic resistance
Resistance genes are often on plasmids or transposons that can be transferred
between bacteria
Misuse of antibiotics selects for resistance mutants
Misuse includes:
Using outdated or weakened antibiotic
Using antibiotics for the common cold and other inappropriate conditions
Using antibiotics in animal feed
Failing to complete the prescribed regimen
Using someone elses leftover prescription

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1. Enzymatic destruction
or inactivation of the
drug
2. Prevention of
penetration to the
target site within the
microbe
3. Alternation of the
drugs target site
4. Rapid efflux (ejection)
of antibiotic
5. Variations of
mechanisms of
resistance
Check Your Understanding 20-17
Effects of Combinations of Drugs
Synergism occurs when the effect of two drugs together is greater than effect of
either alone (W.Win)
Antagonism occurs when the effect of two drugs together is less than the effect
of either alone(V.S.)
(Refer to fig 20.23)
Antibiotic Safety
Therapeutic index: risk versus benefits
Superinfection
Suppression and Alteration of the Microbiota by Antimicrobials
Biota
Normal microbial colonists of healthy body surfaces
Harmless or beneficial bacteria
A few may be pathogens
Broad-spectrum antimicrobials destroy health biota along with pathogens
Superinfection:
Normal resident biota destroyed
Microbes that were once small in number overgrow

Examples of Superinfection
Urinary tract infection caused by E.coli treated with antibiotics
UTI broad-spectrum cephalosporin
Lactobacilli in Female vagina killed by the
Overgrowth of Candida albicans vaginal yeast infection or oral thrush
Antibiotic-associated colitis
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Oral therapy with tetracyclines, clindamycin, and broad-spectrum penicillins

kills off normal biota of the colon
Overgrowth of Clostridium difficile invades intestinal ling releases toxins
diarrhea, fever, and abdominal pain

The Role of Antimicrobials in Superinfection

Primary Sites of Actions of Antimicrobial Drugs on Bacterial Cells

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