INTRODUCTION

I.

Definition, Classification and Characteristics of Microorganisms.

II.

Concept: A kind of microorganism which is small bodies, simple structure, not visible by the
naked eyes, visible usually only with light microscope or electromicroscope.
Classification:
1. Non-cellular type: Virus.
2. Prokaryotic group: with primitive nuclei, e.g. bacteria, actinomyces, rikettsia, chlamydia,
mycoplasma, spirochete.
3. Eukaryotic group: e.g.. Fungi .
Characteristics
1Very small bodies and simply structure.
2To multiple rapidly and mutant easily.
3 Many species of microbes, and distributed widely.
Relation between Microbes and Human---Be more beneficial than harmful to Human
1. the cycle of natural materials
2.industrial and agriculture production
3.medical industry.
4.genetic engineering.
5.normal flora.
6.pathogen and opportunistic pathogen

III.

Developmental history of microbiology

Experiential period of microbiology
Experimental period of microbiology
1. Morphological period
2. Physiological period: Louis Pasteur; Robert Koch; Iwanovsky
Modern Microbiological period.
IV.
BIOFILM -- Community of microorganisms irreversibly attached to solid surface or each
other and encased in an exopolysaccharide matrix.
Circulating surface conditioning
Primary adhesion
Formation of micro colonies
Mature biofilm (lots of bacteria)
Detaches from biofilm, moves to other places to form new biofilm.
Bac close to surf easy to contact with antibiotics
o Inner bac impossible to kill
o DNA of inner bac easy to exchange because lack of nutrients (Die releases bac
frag)
V.
MEDICAL ECOLOGY: A science to study the relation between the microorganisms,
the microorganism and human
body, microorganism , human body and environment. Include: eubiosion dysbiosion
ecological adjustment.
NORMAL MICROBIAL FLORA
o Relation between species :
Symbiosis : neutralism, commensalism, synergism, antagonistic
symbiosis, amensalism
Parasitism
1

o Concept:
inhabit the skin and mucous membranes of healthy normal persons
nonpathogenic
can play a definite role in maintaining health and normal function
o Distribution: Skin; Intestinal tract; Respiratory tract; Genitourinary tract
o Physiologic functions of normal flora:
1) Bacterial interference
competition for receptors or binding sites
competition for nutrients
antibiotic materials or bacteriocins
alter PH
2) Nutritious function
synthesize vitamin K and vitamin B
aid in the adsorption of nutrients.
3) Immunostimulation
induce sIgA
increase the activity of mononuclear-phagocytes
increase the releasing of cytokines
4) Inhibition the growth of tumor cell
5) Resistant to the old and feeble
o Normal flora of the body
the role of the normal mouth flora in dental Caries
normal floral of the genitourinary tract
normal flora of the intestinal tract
Esophagus and stomach
Small intestine
Large intestine (pathogen; symbiosis; Intermediate)
normal floral of the respiratory tract
normal floral of the skin
Microeubiosis and Microdysbiosis
o Microeubiosis: this is a dynamic balance
Related factor: location; character; quantity; host
Evaluate: from host; from microorganism
o Microdysbiosis
1) Classify
Flora disequilibrium:
I degree
II degree chronic disease
III degree dysbacteriosis (superinfection)
Shift of location
Metastasis through blood circulation
Focus which alter location
2) Induced factors: irradiate of ray; usage of antibiotic; surgical operation; others
o Prevention & control of Microdysbiosis:
protect the environment; increase the immunity
make rational use of antibiotic
principle: just the right amount; specificity; no-oral giving drug;
protecting anaerobe
usage of microecological preparation
2

 Opportunistic Infection
I. Opportunistic bacterium : normal flora opportunistic bacterium
specific condition: flora disequilibrium; shift of location; decrease of hostimmunity
II. Opportunistic infection
CHAPTER 1 : BASIC CHARACTERS OF BACTERIA
I.

Bacterial Morphology and Structure

Bacterial Morphology
Units of measurement: m (micron or micrometer); nm;
Basic morphology: Coccus; Bacillus; Spirillar bacterium; (Vibrio; Spirillum;)
Arrangement of bacteria: Staphylococcus, diplococcus, streptococcus, tetrad,
sarcina, coccobacillus, streptobacillus, vibrio, spirillum.
II. Basic Structure Of Bacteria
1. Cell Wall
1) Gram-Positive Bacteria(G+):
PEPTIDOGLYCAN
back-bone: N-acetylmuramic acid(M)
N-acetylglucosamine(G)
-1.4glycosidic
side chain(tetrapeptide)
peptide cross-bridge:Ala-Glu-Lys-Ala
TEICHOIC ACID: wall teichoic acid; membrane teichoic acid

2) Gram-negative bacteria(G-):
PEPTIDOGLYCAN
back-bone: N-acetylmuramic acid(M)
N-acetylglucosamine(G)
-1.4glycosidic
side chain(tetrapeptide)
peptide cross-bridge:Ala-Glu-Lys-Ala
OUTER MEMBRANE
3

LPS

lipid protein
phospholipid bilayer
lipopolysaccharide(LPS)
lipid A
core polysaccharide
specific polysaccharide

2) Comparison Of Cell Wall For G+ And G_:

COMPONENT
G+
THICKER
PEPTIDOGLYCAN
TEICHOIC ACID

YES

GTHINNER
NO

LIPOPOLYSACCHARIDE NO

YES

PNC

ANTIGENICITY

TECHOIC ACID

LPS

Gram Positive
Teichoic acid
Wall teichoic acid
Membrane teichoic acid
Peptidoglycan
50 sheets
Tetrapeptide side chain : Ala-Glu-LysAla
Backbone: Alternating Nacetylglucosamine and N-acetylmuramic
acid subunits connected with glycosidic
bonds
Pentapeptide cross bridges

Gram Negative
Peptidogylcan
Thin layer, 2-3 sheets
Doesnt have pentapeptide bridge
DAP AA instead of Lys
Outer membrane
Lipid protein
phospholipid bilayer
lipopolysaccharide (LPS)
Lipid A
Core polysaccharide
Specific polysaccharide

 Penicillin
Tetrapeptide and pentapeptide connecting
point sensitive to.
Prevents cell wall from forming
3) Function Of The Cell Wall
maintain the shape of bacteria
protect from low osmotic pressure
permeability
surface antigenic determinates
pathogenicity
4) Main component: Peptidoglycan (i.e. mucopeptide):
Backbone ; Side chain; (Gly)5 peptide cross-bridge
III.
L Type bacteria ( Bacteria L form ) : Cell membrane ; Cytoplasm; Nucleus;
Mesosome, Chondroid.
Characteristics :
lacking of the bacterial wall
protoplasts from gram-positive bacteria spheroplasts from gram-negative bacteria
lacking a rigid cell wall-regular size and shape
difficult to cultivate and require a medium
that is solidfied with agar as well as having
the right osmotic strength
some can revert to the normal bacillary
L-forms can produce chronic infection
Structure :
Cell membrane
Structure: a typical unit membrane, which composed of a phospholipid bilayer and
proteins similar to that in eukaryotic cells, but do not contain sterols.
Function:
Selective permeability and active transport
Energy generation
Synthesis of precursors of the cell wall
Secretion of hydrolytic exoenzymes and toxins
Mesosome(Chondroid)
Structure: Septal mesosomes; Lateral mesosomes
Function:
Compartmenting of DNA at cell division and at sporulation
Providing a membranous support for respiratory enzymes
Convoluted invaginations of cell membrane
Acts on an anchor to bind and pull apart daughter chromosomes during cell
division (like EU spindles)
Increases membrane area --> enzyme content and energy production enhanced
Cytoplasm
1) Ribosome --- Being the site of protein synthesis

50S subunits
70S

60s
80S

30s subunits
40s
(bacterial)
(eukaryotic)
2) Inclusion granules
Function :
----serve as storage areas for nutrients
----consist of volutin ( polyphosphate ),
lipid, glycogen, or starch
----metachromatic grandules
characteristically found in the Diphtheria bacillus
Having distinguishable significant
3) Plasmids
Concept: extrachromosomal , closed double-stranded, circular DNA molecules
Important plasmid on medicine
F plasmid----Sex pili
R plasmid----antibiotic resistance
Vi plasmid--- several toxins
Col plasmids---bacteriocins
Other plasmids---a variety of degradative enzymes
Characteristics
self-replicating independently of the bacteria chromosome
Be not essential for bacterial survival, can be absent by themselves
can be transferred from one to another bacterial cell by conjugation or
transduction
Plasmids carry genes associated with specialized functions
4) Nucleoid (nuclear material)
The area of cytoplasm in which DNA is located
Contains no nuclear membrane, no nucleolus, no mitotic spindle, and histones
They are bacterial genetic material
Special structures of bacteria
1. Capsule
Concept
Contact with the cell wall immediately.
thick enough ( 0.2m or more )
Chemical composition
Polysaccharide or polypeptide
Development of it is often dependent on favourable environmental condition.
Function
Protect the cell wall against attack by various kinds of antibacterial agents
play a role in the adherence of bacteria to human tissues
play a very important part in determining the antigenic specificity of bacteria
Examination: capsule stain and negative stain
2.Flagella
Concept: Only certain bacteria have flagella:
--- all spirochetes and many rods --- but most cocci without it
Four types of arrangement
Monotrichate, amphitrichate,
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IV.

Lophotrichate and peritrichate

Composition: Protein (flagellin)
Basal body
Structure Hook
Filament
Function: motility; antigenicity
Examination
The flagella stain
Observe directly by EM
Observe the motility of bacteria
Inoculating on the semisolid agar
3. Pili ( Fimbriae )
Concept
Many G- bacteria and few G+ bacteria possess
rigid surface appendages
shorter and straighter than flagella
Composition: composed of subunits of a protein----pilin
Classes And Function
1. Ordinary pili
--- adherence of pathogenic bacteria to host cells
--- being the site of the major surface antigen
2. Sex pili
--- only appear in a few G- bacteria
--- having 1~4 being borne per cell
--- sex pili are coded by F plasmid
--- mediating bacterial conjugation
--- acting as receptor sites for certain bacteriophages
Examination: Observed through EM
4. Spores ( endospores )
Concept
Characteristics
a highly resistant resting phase
spore formation occurs when nutrients are depleted
forming insides the cell
containing necessary components for bacterial life
resistance of the spore may be mediatedby dipicolinic acid
having low metabolic activity and remaining dormant for many years
Relation between spore and vegetative forms
Germination
spore
vegetative forms
Sporulation
Examination: The spore stain
Significance in medical science
Having extraordinary resistance to heat and chemicals
can cause diseases through germination
Having significance in differentiation
Examination and Staining:
1.Simple staining
2.Complex staining
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Grams stain
differential staining
Acid-fast stain
Capsule stain
special staining
Flagella stain
Spore stain
Procedure of Grams Stain; Practical Significance of Grams stain:
First stain
Crystal violet solution , 1
wash
Mordant
Lugols iodine solution , 1
wash
Decolorize
95% alcohol for 30
wash
Counter stain Dilute carhol fuchsin , 1
wash and check
3. Results
violet ----- G+
Red ----- G4. Practical significance: Differentiate bacteria; Selecting drugs; Judge the pathogenic ability
5. Consists of: Primary stain (violet gram dye); Iodine ; 95% alcohol ; Saffron
V. Bacterial Metabolism and Multiplication
1) Chemical Composition and Physical Characteristics of Bacteria. (Self-learning)
2) Growth and Mutiplication of bacteria
1. Nutritional typies of bacteria: autotrophs; heterotrophs
Bacteriophage
Viruses which can infect bacteria, fungi and spirochetes
Highly host specific
Morphology
Measurment: nm (EM)
Consists of: head, tail
Lytic Cycle
Multiply process of virulent phages
Adsorption, penetration, biological synthesis, maturation, release
3) Basic requirement of bacterial multiplication
Nutrients: Water; Carbon Source; Nitrogen Source; inorganic salts; growth
factors
Optimal pH : 7.2-7.6
Special for vibrio: 8.5-9
Proper Temperature : 370C
Definite gaseous (aeration): O2, CO2, N2
Obligate aerobes: T.b;
Cholera
Microaerophilic bacterium: C.jejuni
Facultative anaerobe: the most bacteria
Obligate Anaerobes: C. Tetani
o Anaerobic principles of anaerobes
(1) Do not possess cytochrome oxidase nor cytochrome.
(2) Do not possess hydrogen peroxidase, catalase and superoxide dismutase (SOD).
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o Methods and speed of bacteria growth.

1. Growth Of Individual
Manner ---simple binary fission
Speed ---One generation/15~20 for general
One generation/16~18hr for T.b
2. Typical growth curve(growth of population)
Lag phase --- a period of adaptation
Logarithmic phase --- rapid growth. Observe bacterial biological
property
Stationary phase --- Gain bacterial metabolite and spore
Decline phase

4) Cultivation of bacteria
5) Artificial culture of bacteria
Culture media:
Concept:
Classification:
According to physical condition

According to nutriental composition

Liquid medium
Semisolid medium
Solid medium

Basic Medium
Enrichment Medium
Selective Medium
Differential Medium
Anaerobic Medium
The phenomenon of bacterial growth
Physical condition
Liquid medium-- turbid, sediment, gas producing
Semisolid medium-- motility
Solid:
slant (bacterial storage)
Plate (pure cultures- colonies smooth, rough, mucoid
Nutrient composition: Basic, enrichment, selective, differential, anaerobic
culture medium
6) Metabolism of bacteria
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VI.

VII.

1. Biochemical reaction of bacteria

SUGAR : Sugar fermentation of bacteria; Methyl red test (M )
AMINO ACID:Indol test (I); Hydrogen sulfide production test
CITRATE UTILIZING TEST (C)
Significances of Products of medical importance (synthetic products):
Pyrogen :
Toxins and enzymes : exotoxins
endotoxins
enzymes of invasiveness
Antibiotics:
Bacteriocin:
Vitamin: Vitamin B and K
Pigment: Possessing differential significance
Morphological identification of bacteria
1) Examination of unstained specimen:
2) Examination after staining
simple staining:
Grams stain
complex staining
Acid fast stain
Special stain
GRAMS STAIN
Principle of Grams stain: structure of cell wall
crystal violet solutions
2. Step of Grams stain:
(1) First stain
iodine solution
(2) Mordant
(3) Decolorize
95%alcohol
(4) Counter stain
diluted carbol fuchsin

CHAPTER 2 BACTERIAL GENETICS AND VARIATION

I. DEFINITION
o Heredity
o Variation Genetical Variation Change in genotype
Non-genetical Variation Change in phenotype
II. VARIATION OF BACTERIA
Variation in morphology and structure: lose capsule (Pneumococcus); H O; Lose
Spore; L-form
Variation in virulence

Bovine TB BCG (13years, 230 generation)

C. diphtheriae : b-corynephage, diphtheriae toxin
Variation in drug resistance: Penicillin resistant strains of staphylococcus aureus
Variation in colony: S-R
Variation in enzyme activity: Phenotypic and Genotypic
III.
PHENOMENON OF BACTERIAL VARIATION
1. Morphological And Structure Change
on media
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Capsule

lose of capsule
in mouse
0.1% carbolic acid

Flagella
H

lose of Flagella
common media
O
PNC & lysozyme
Bacteria
L form bacteria
without
42 10-20days
Anthrax Bacilli
Lose of spores
Attenuated Virulence
2. Variation of virulence
Glycerine potato
Bovine tubercle
BCG
bacillus
13 years,
Without virulence
230 generations
-corynephage
Diphtheria bacilli
Without virulence

Gaining virulence
Lysogenic conversion

3. Antibiotics resistant variation

4. Colonial Variation (SR)

IV.
GENETIC MATERIALS OF BACTERIA
1. Chromosomes : DNA forms: ds-DNA, circle
Size : E.coli 1300mm, 4288gene; Rolling-circle pattern of replication

Replication

DNA

RNA

Transcription

Protein

Translation

2. Plasmids: Extrachromosomal genetic elements that are capable of autonomous

replication.
Small double-stranded DNA molecules, usually circular
exist independently of host chromosome
autonomously replicating (replicon)
may disappear spontaneously or by induction (UV)
11

 incompatibility and compatibility

Classification of Plasmids
Transfer properties
Conjugative: 40-100kbp; eg. F, R plasmid
Nonconjugative : <15kbp, transfer by mobilization; eg. ColE1 plasmid
Phenotypic effects
Fertility plasmid,F plasmid: coding sex pilus
Resistance plasmid, R plasmid: resistance transfer factor; resistance
determinant
Virulence plasmid: Coliciogenic plasmid
3. Bacteriophage / Phage
Definition
(1) one kind of viruses that could infect bacteria, fungi, spirochaete et al.
(2) multiplication in living host cells
(3) being highly host-specific
(4) can lyse the host cells
Properties of phage
Morphology: Unit of measurement - nm; Visible with EM
Other characteristics --- antigenity, resistance
Composition and structure

Composition
o Nucleic acid: DNA or RNA
o Protein: Protection, Infection

Structure (T4): Head or capsid; tail

Types of Bacteriophage (Relation between host cell and phage)

Lytic or virulent phage (e.g., T4): Phage that can only multiply within
bacteria and kill the cell by lysis. (e.g., T4)
Produce many copies of themselves as they kill their host cell
Replicative cycle:
Adsorption Penetration
Biosynthesis
Assembly
Lysis and release

Lytic cycle :
Attachment
Injection and uncoating
Biosynthesis
Eclipse: Early proteins; Phage DNA synthesis; Late proteins
Intracellular accumulation
Maturation and releasing

12

Lysogenic phage
Lysogenic or temperate phage(e.g.): Phage that can either multiply via
the lytic cycle or enter a quiescent state in the bacterial cell. (e.g., l)
can integrate their genomes into those of their host cells
can produce lysogenic state
Prophage --- phage DNA that exists and replicants
along with the bacterial DNA
Lysogenic bacterium --- A bacterium that carries a prophage.
Lysogenic state --- Some phages are able to infect a bacterium
without inducing the production of more phage or the lysis of the
infected cell, and integrate their genome into that of the host
cell, phage DNA replicates along with the bacterial DNA.
Artificial stimuli may cause depression of prophage, and stop the
lysogenic state.
Lysogenic or phage conversion: A change in the phenotype of a bacterial
cell as a consequence of lysogeny
Modification of Salmonella O antigen
Toxin production by Corynebacterium diphtheriae

13

Infection of Host Cells

Attachment Sheath contraction

Nucleic acid injection

Application of phage
Diagnosis
Identification and typing bacteria
Genetic engineering
As a vector
Transduction
Phage Conversion
4. Transposable elements
Concept: are pieces of DNA that move readily from one site to another,within
between the DNAs of bacteria ,plamids and bacteriophage.
Function : mutation, drug resistance enzymes, toxins
Properties: Random movement; Jumping genes or movable genes; First
discovered in the 1940s by Barbara McClintock during her study on maize
genetics.(won the Nobel prize in 1983).
Classification :
(1) Insertion sequences (IS): A short sequence of DNA containing only the
genes for those enzymes required for its transposition. Importance:
mutation and plasmid insertion.

*IR : Inverted Repeat

14

(2) Transposons (Tn) or complex Tn : contain genes other than those

required for transposition (eg. Antibiotic resistance or toxin genes).
Importance : antibiotic resistance.

V.

(3) Mutator bacteriophage, Mu phage

MECHANISM OF BACTERIA VARIATION
Mutations in bacteria
spontaneous mutation
induced mutation
point mutation
multiple mutation
Character
occur with a frequency of 10-8~10-6 but can be induced by artificial
stimuli
posses spontaneity and random
reverse mutation
Wild strain
Mutant strain

Changes in DNA sequences : Base substitutions, deletions, insertions,

rearrangements
Backword mutation or reverse mutation
Gene transfer and recombination
General Features of Gene Transfer in Bacteria

Unidirectional: Donor to recipient

Donor does not give an entire chromosome

Gene variation can occur between species

1. Transformation : Gene transfer resulting from the uptake of DNA from a
donor. Competence of the recipient (Bacillus, Haemophilus, Neisseria,
Streptococcus).
Cell-free DNA
DONOR

RECIPIENT

Absorb directly
Condition of transformation
Homolog between donor and recipient
Recipient cell depend on their competence (in the growth cycle )
2. Conjugation: Donor DNA transferred to recipient cell through sex pilus.
Conjugative Plasmid
DONOR
RECIPIENT
Sex pili
Character --- Contact of the two cells directly
Method --F+ F - F+
Hfr F - FF plasmid
R plasmid--- component of RTF(coding sex pili)
15

Resistance determinant
Function : relate with mutiple antibiotic resistance
TYPES
INFORMATION
F PLASMID (F+ F - F+ )
Mechanism of F+ x F- Crosses
Pair formation : Conjugation bridge
DNA transfer: Origin of transfer;
Rolling circle replication

Physiological States F plasmid

F plasmid Hfr (High Frequency Of
Recombination)

F plasmid

Mechanism of Hfr x F- Crosses

16

Mechanism of F x F- Crosses

Resistance plasmid, R plasmid

RTF (resistance transfer factor)
o transfer genes
o code pili
R determinant(resistance determination)
o resistance genes
o transponsons

3. Transduction : Gene transfer from a donor to a recipient by way of a

bacteriophage. Types :
Generalized transduction any of donor genetic information transfer to the
chromosome of recipient. Example :

Infection of Donor

Release of phage

Phage replication and

Infection of recipient
degradation of host DNA

Legitimate recombination

Assembly of phages particles

17

Restricted transduction Bacteriophage can only transfer certain genes

from the donor to recipient cells.

Plasmid, chromosome
DONOR

RECIPIENT
Temperate phage

4. Lysogenic conversion Some bacteria are infected by a temperate phage and

their DNA recombinated together and expressed
18

Prophage DNA
recipient
5. Protoplast fusion
6. DNA Recombination
VI.
Practical implications
1. Application in diagnosis, treatment and prevention of infectious diseases. Eg. L-form;
PCR
2. Detection of mutagenicity The Ames test.
3. Application in genetic engineering
CHAPTER 3 : BACTERIAL DRUG-RESISTANCE
I.

Introduction
o Antibacterial agents and antibiotics
o Antibiotic resistance in human and animal
o Mutiple drugs resistance (MDR)
II. Main mechanisms of antibiotic resistance formation
1. Genetic Mechanisms
1). Intrinsic resistance
2). Acquired resistance
--- Bacterial chromosomal mutation
--- Plasmid mediated Drug-Resistance
--- Transposon ( IS ; Tn ; Mu phage )
2. Biochemical Mechanisms
1). Target site modification
(1) PBPs--- MRSA
(2) DNA gyrase--- gyrA and gyr B gene mutation
2). Production of modified enzyme
(1) -lactamase --- inactivated enzyme penicillinase and cephalosporinase
(2) aminoglycoside-modifiled enzymes
Phosphotransferase
Acetyl transferase
Adenyl transferase
(3) chloramphenicol acetyl transferase
3). Active efflux pump
P. Aeruginosa--MexA, MexB-OprM
MexC, MexD-OprJ
MexE, MexF-OprN
MexX, MexY-OprM
4). Change of permeability of bacterial cell wall
Decided by Porin : E. Coli--- OmpF and OmpC
3. Common drug-resistant bacteria in hospital infection
Pseudomonas aeruginosa --- PA
Mycobacterium tuberculosis
Neisseria gonorrohoeae
Staphylococus aureus --- SA
Vancomycin resistant enterococci --- VRE ; Vancomycin resistant staphylococcus
aureus (VRSA)
4. Prevention and treatment principles
1). Minimal inhibitory concentration --- MIC
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2). Minimal bactericidal concentration --- MBC

Drug sensitivity test
CHAPTER 4 : BACTERIAL INFECTION AND PATHOGENESIS
I.

Bacterial Infection

1. Infectious sources
1) Exogenous infection --- patients ; carrier; animal
2) Endogenous infection --- normal flora; latent bacteria
2. Infectious Types
1. Inapparent infection (subclinical infection)
2. Apparent infection: Toxemia; Septicemia; Bacteremia; Pyemia; Endotoxemia.
3. Transmissible manner
infection through mucosa --- local and spreading
infection through hurting
infection through muti-roads
3. Infectious type
Inapparent Infection --- Subclinical Infection
Apparent Infection
Acute Infection
Chlornic Infection
Local Infection
Generalized Infection
Toxemia
Septicemia
Bacteremia
Pyemia
Endotoxemia
II. Bacterial Pathogenecity
Pathogenecity
Virulence (LD50 or ID50)
Invasiveness
Toxigenicity
Number of bacterial entry
Portal entry
*)
LD50 (Lethal Dose 50) the number of microbes or micrograms of toxin that must be
administered to kill 50% of the animals.
1. Virulence --- toxic factor
1) Invasiveness :
Bacterial related structures
--- pili or LTA
--- capsule or microcapsule
Invasive enzyme
--- Hyaluronidase spread
--- Coagulase anti-human defense
--- IgA protease; streptokinase.
2) Bacterial toxin
o Exotoxin
Origin --- Produced mainly by G+, Liberated from living bacterial cell
Nature --- Protein
Type --- Neurotoxins, Cytotoxins, Enterotoxins
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Character --- Highly toxin; Highly selectivity; Highly antigenity (can produce
anti-toxin); Detoxification.
0.4%formalin
EXOTOXIN
TOXOID
4~6weeks
Molecule structure: A subunit
+ B subunit
o Endotoxin
Origin
--- Produced mainly by G--- Liberated from cell wall of dead bacteria
Nature
--- LPS
Character --- Weakly toxin; Having no selectivity; Weak antigenity (can
not produce toxoid); Having no detoxification.
Structure of LPS
lipid A
corepolysaccharide
specific polysaccharide
Biologic effects of bacterial toxin
Fever due to the release of endogenous pyrogen
Increase of numbers of WBC
Shwartzman and DIC
Hypotension, shock, and impaired perfusion of essential organs
Activation of the complement, resulting in inflammation and tissue damage
Immunoregulation can be caused by less endotoxin
DIFFERENTIATION ON ENDOTOXIN EXOTOXIN
PROPERTY
ENDOTOXIN
EXOTOXIN
Certain species of
Cell wall of most GSOURCE
some G+and GSECRETED FROM CELL
Yes
No
CHEMISTRY

Polypeptide

Lipopolysaccharide

LOCATION OF GENES

Bacterial chromosome

TOXICITY

Plasmid or
bacteriophage
High

CLINICAL EFFECTS

Various effects

Fever, shock

ANTIGENICITY

Induces high-tiler
antitoxins
Toxoids used as vaccines

Poorly antigenic

VACCINES

Low

No toxoids formed

Destroyed rapidly
Stable at 1000C for 1 hour
0
at 60 C
Pathogenicity of virulent factors --- Superantigen; Pathogenicity island
2. Number of bacterial entry
3. Portal entry
HEAT STABILITY

HOSPITAL INFECTION (NOSOCOMIAL INFECTION)

21

 Conception: Nosocomial infection are those that arise during hospitalization as a

comlication of another illness. e.g. acquired in hospital.
I.
Classification According To The Sources Of Pathogens:
1. Endogenous nosocomial infection (self-infection): Colonization changed; lower
immune function; dysbacteriosis.
2. Exogenous nosocomial infection (cross-infection): patients; carrier; environment;
animal.
II.
Classification According To The Infection Position: urinary drainage infections
most common; surgical infections; skin infection;
III.
Common Properties Of Pathogenic Microbes
Conditional pathogen.
Drug-resistant bacterium.
New pathogen.
Higher resistance to environments.
IV.
Prevention And Control
1. Asepsis: sterilization and disinfection; operation room; hand wshing; person
2. Isolation precaution : patients; drainage-secretion; blood mainpulation.
3. Antibiotics application in rationality
4. Organization: surveillance network.
main principles: prevent disease; reduce the costs of medical care.
CHAPTER 5: IMMUNE DEFENCE AGAINST INFECTION
(Human Immunity against Bacterial and Viral Infection)
I.

Nonspecific immunity
Biological Barriers
Phagocytosis
Humoral Factors
II. Specific immunity
o Cellular and Humoral immunity
o Immunity against extracellular bacterium -- Neutrophil, Ab, complement
o Immunity against intracellular bacterium -- Macrophage, Sensitized T, CTL
o Immunity to exotoxin -- Antitoxin, SIgA
III. Antiviral immunity
Non-Specific Immunity (Innate resistance)
Specific Immunity
Synergism of immune factors
Duration of acquired immunity.
CHAPTER 6: LABORATORY DIAGNOSIS OF BACTERIAL INFECTION
I.

Procedure of detection for pathogenic bacteria

Sample collection and treatment : Smear and stain sample (primary diagnosis)
Cultivation and isolation
Biochem test
Serological test / Identification (detection of Ab)
Animal test
Drug sensitive test
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II.

Phage typing
Molecular technology : Fingerprinting, DNA hybridization; PCR
Serological diagnosis : For detection of Antibodies.
Prevention (control) and treatment: Enhance the immunity (resistance) of human
body
CHAPTER 7: PREVENTION OF PATHOGENIC MICROBIAL INFECTION

I.

INTRODUCTION
Disinfection : Killing pathogenic microorganism, meaning vegetative forms
not spore.
Sterilization : Killing or removal of all microorganism, including bacterial
spore.
Bacteriostasis : bacteriostat used to inhibit microorganisms in vivo or vitro.
Antisepsis : chemicals used to inhibit or kill microorganisms in vitro.
Asepsis : not including living bacteria
II. PHYSICAL AGENTS
1. Heat
Mechanism : denaturing proteins; membrane or wall damage; enzymatic
cleavage of DNA
Classification :
dry-heat : incineration ; Hot-air oven
moist-heat

Pasteurization---be used primarily for milk

Boiling --- mainly for vegetative forms; 1000C within 2-3minutes

Fractional sterilization

Autoclaving -- most frequently used method of sterilization; pressure

of 15 1b/in2, temperature of 1210C; for 15~20 minutes; Killing all
microorganism, including spore
2. Radiation
(1) ultraviolet (UV): 250~260nm wavelength region of maximum
absorption by DNA formation of thymine dimer, DNA replication is
inhibited for air and surface of subjects can damage cornea and skin
(2) X-ray
3. Filtration
o for sterilizing certain solutions with heat-sensitive components
o physically trapping particles larger than the pore size
4. Ultrasonic vibrations
5. Drying
6. Cold lyophlization ( freeze drying)
III.
CHEMICAL AGENTS
Mechanism
o disruption of the lipid-containing cell membrane
o modification of proteins
o modification of DNA
1. Common disinfectants
Phenols carbolic acid, Lysol
Halogens iodine
Alcohol ethyl alcohol
Aldehyde formaldehyde,
23

glutadehyde
Acids and alkalies acetic acid,
lime
Surface active substances
1. Mechanism of chemical disinfectants
Cell membrane injury
Coagulation and denaturation
Interactions with functional groups of proteins
3. Factors influencing chemical disinfectants
Concentration
Time
Organic materials
pH
Temperature
Kinds of bacteria
4. Affected agents of disinfection
Disinfectant : type, concentration, time
Bacteria : age, type
Environment : organism, Ph, temperature
5. Selection of disinfectant: low toxicity; cheap; low destroy ; easy to store;
convenient; effective

I.

II.

III.

IV.

CHAPTER 8: VIRAL PROPERTIES AND PATHOGENESIS

Concept of virus:
Viruses are the smallest infectious agents and contain only one kind of nucleic acid
(RNA or DNA) as their genome.
The nucleic acid is encased in a protein shell,which may be surrounded by a lipidcontaining membrane.
Viruses are inert in the extracellular environment; they replicate only in living cells,
being parasites at the genetic level.
The entire infectious unit is termed a virion
o Having intactness of structure
o Having property of infectiousness
o Being a structure of outside-cellular
Nature of virus
Smallest
either RNA or DNA, surrounded by a protective protein shell
Non-cellular type of microorganism
Reproduce within cells obligate intracellular parasites
Resistant to antibiotics, sensitive to IFN
Classification of virus
According to transmission route: Virion; Physicochemical Property; Virus
genome; Virus proteins; Genome organization and replication; Ag prop; Bio
Prop
Shape of Virus: Spherical; Rod; Brick; Tadpole; Bullet; Filament
Many infections diseases caused by viruses
Fatal --- rabies, smallpox, HIV etc
Contagious --- influenza, measles etc
Congenital abnormalities --- CMV, HSV etc
Tumors and cancers --- HBV,HPV,EB etc
RABIES
Rabies is a disease caused by the rabies virus. It may take several weeks or even
a few years for people to show symptoms after getting infected with rabies, but
24

usually people start to show signs of the disease 1 to 3 months after the virus
infects them.

V.

VI.

The early signs of rabies can be fever or headache, but this changes quickly to
nervous system signs, such as confusion, sleepiness, or agitation. Once
someone with rabies infection starts having these symptoms, that person usually
does not survive.
SMALLPOX
Smallpox is a serious, contagious, and sometimes fatal infectious disease.
There is no specific treatment for smallpox disease, and the only prevention is
vaccination.
Smallpox outbreaks have occurred from time to time for thousands of years,
but the disease is now eradicated after a successful worldwide vaccination
program.
After the disease was eliminated from the world, routine vaccination
against smallpox among the general public was stopped because it was no longer
necessary for prevention.
HPV
A human papillomavirus (HPV) is a papillomavirus that infects the skin
and mucous membranes of humans.
Persistent infection with "high-risk" HPV types may progress to precancerous
lesions and invasive cancer. HPV infection is a cause of nearly all cases of
cervical cancer.
HPV is the most common sexually transmitted infection in the United
States. Estimates of prevalence vary from 14% to more than 90%.
The Basic properties of Viruses
Size and morphology of virus
Size --- Unit of measurement -- nm
Visible with EM
largest---Poxvirus
300X250nm
smallest---Polio
30nm
Mk --- spherical & others
The most common viral morphologies. A naked icosahedral virus (e.g.
poliovirus), an enveloped icosahedral virus (e.g. herpes virus), a naked helical
virus (e.g. tobacco mosaic virus) and an enveloped helical virus (e.g. influenza
virus). Individual capsomeres are arranged to form a capsid which encloses the
nucleic acid (DNA or RNA) of the virus. Many animal viruses also contain an
envelope, which is partly derived from the host cell membtrane but which
always contains unique viral proteins drawn here as "spikes".
Structure and components of virus
Nucleocapsid =Nucleic acid +capsid
Virion = Nucleocapsid (Naked virus) = Nucleocapsid + Envelope (envelope
virus)
Structure
1. core : nucleic acid genome
Deciding the heredity, variation and replication of virus
2. capsid :
POLYPEPTIDE CAPSOMERE

CAPSID
(structural sub)
(morphologic sub)

25

protecting , introducing and antigenity

Arrangement: Helical Symmetry; Cubic Symmetry; Complex Symmetry
capsid + genome nucleocapsid (Or naked virus)
3. envelope
PEPLOMERE

ENVELOPE

protecting , introducing and antigenity

surrounding the nucleocapsid
acquired as budding through host cell
peplomere or spike on the surface
nucleocapsid +envelope enveloped virus

Component
1) Nucleic acids

DNA or RNA

double-strand or single-strand; linear or circular

Mw: 2~200X106; Bp: 2~200X103

ORF (open reading frame )

containing all genetic information
Roles --- taxonomy
infectious nucleic acid
2) Proteins

Nature --- structural or non-structural proteins

Analytic methods --- SDS-page

Roles:
protecting the genomes
functional proteins, eg.
mediate the attachment
enzymes
determining the antigenicity
having self-toxicity
3) Lipid

Roles --- protecting; causing fever, toxicity; mediate the attachment

VII. Viral Multplication
Definition
1. Viral multiplication --- self replication
2. Multiplicative cycle: Adsorption and penetration; uncoating; biosynthesis ;
assemble and release
Replication cycle
1. Adsorption
26

1) Ionic attraction : at pH7.0; non specific; reversable

2) Receptor combine: specific; non reversible; e.g. HIV---CD4, HBV--PHSA
2. Penetration and uncoating: Often occur simultaneously
1). Penetration | Model--- injection; viropexis; membrane fusion
2). Uncoating : hydrolysis----- envelope; enzymes ----- capsid
3. Biosynthesis
1) Replication of genome | Principle --- semi-conservative replication

DNA --- ds and ss

RNA --- ds; +ss; -ss; retrovirus

2) Synthesis of proteins

parent genome early mRNA early proteins

(functional)

progeny genome late mRNA late proteins (structural)

4. Assemble and release of progeny virus
1) Assemble
viral genome + capsid proteins progeny virus particles
DNA virus- nucleus
location
RNA virus-cytoplasm
2) Release
naked virus --- cell lysis
enveloped virus--- budding
STAGES OF THE VIRAL GROWTH CYCLE
Attachment and penetration by parental virionUncoating of the viral
genomeEarly viral mRNA synthesisEarly viral protein synthesisViral genome
replicationLate viral mRNA synthesisLate viral protein synthesisProgeny
virion assemblyVirion release from cell
VIII. Abnormal Multiplication
1. Defective virus

containing normal capid proteins

having mutation or a deletion of part of genome

reproducing with helper virus

interfering the homologus virus

2. Abortive infection --- the virus enters the non-permissive cell which can not
provid the essential enzymes & energy to viral multiplication permissive cell and
non-permissive cell
IX.
Viral interference
Concept
Probably mechanism

inducing interferon (IFN)

blocking or destroying receptors for virus

altering the metabolic pathway of host

emerged defective virus (DIP)

Significances

Benefits --- breaking off disease infected by virus

Disadvantage --- interfere with effect of inoculation of vaccine

Complementation & Phenotypic mixing

27

VIRUS A

VIRUS B

Phenotype
mixing

VIRUS A

VIRUS B

Integration
INACTIVATION OF VIRAL
o Affected by physical agent : Heat; PH; Radiation
o Affected by chemical agent :
Mechanisms

Disruption of the lipid-containing envelop

Modification of proteins

Modification of DNA
XI.
SUBVIRUS
Satellite Virus (E.G.Virusoid)
Viroid
Prion
o an infectious factor of protein with no detectable nucleic acid
o Basic structural unit is prion protein
o PrPc PrPsc while obtaining pathogenicity and transmissibility
o possesing strong resistant to various factors such as heatproteinase K
ultraviolet et.
o Cause TSE of animal and people, such as Kuru, Creutzfeldt-Jakob disease(CJD),
Scrapie, BSE et
XII. VIRUS INFECTION AND IMMUNITY
Infections pattern and pathway
1. Horizontal transmission --- from each other of groups
1) Via mucous membrane

respiratory ---- surface infection

gastrointestinal----naked virus

genitourinary --- occur with sexual or newborn delivery

2) Via skin

injection-------HBV, HIV

bite -------- rabies virus; damaged skin

2. Vertical transmission

from parents to progeny

more than 10 viruses

the most common: Rubella, CMV, HBV, HIV

X.

28

3. Spreading within the host

Local (superficial ) spreading

Generalized spreading by blood, lymph and nerve fiber

Types of infection
Virus body inapparent viral infection
acute viral infection
persistent viral infection
1. Inapparent viral infection

Virus body asymptom

Reasons --- immunity viral virulence

2. Acute viral infection

Virus body symptom

Results--- tissue damage; toxic products

3. Persistent infection

Virus body persisting a long period

Reasons :
o weak immunity
o weak Ag
o abnormal replication or DIP interfere
o integration
1) Chronic infection

chronic and in progress

course of illness 1 year

e.g. chronic hepatitis cirrhosis cancer

2) Latent infection

recurrent or repeatedly onset

e.g. varicella virus

3) Delayed infection (slow virus infection)

long incubation period; bad prognosis

e.g. SSPE, Kuru

Mechanism of viral pathogensis
1. Effect of virus infection on host cell
1) Cytocidal effect
inhibition of macromolecular biosynthsis
lysosome damage self lysis
cell organella damage
change of Antigenity
cytotoxicity of viral protein
Results--- cytopathic effects(CPE)
2) Steady state infection
appearance of new antigenic determinants
on the cell surface cell fusion
eg. Influenza and hemagglutinin
3) Cell apoptosis
4) Integration of virual gene
5) Cell transformation and malignant --- cancer
6) Formation of inclusion body
2. Effect of virus infection on host
1) Direct damage --- eg. Pollo. V, motor neuron
29

2) Immunopathology system --- Type II, III and IV hypersensitivity

3) Direct effect on immune system
(1) Immune suppression --- HSV, Measles virus, Humor Immunity
(2) Immune competent cells--- HIV, CD4+ cells
(3) Auto-immune diseases
CHAPTER 9: COCCI
I.

Common features and classification .

Grouped together based on

Gram stain reaction

Thick cell wall composition

Spherical shape
Classification of Pyrogenic Coccus :
G+: Staphylococcus, Streptococcus
G-: Meningococcus, Gonococcus
Common: Pus; Enzyme and toxin production
II.
Comparison between staphylococcus and streptococcus .
Staphylococcus
.Biological Properties:
Morphology And Culture : Spherical cocci , G+; arranged in grapelike clusters, Nonmotile,
dont form spores; Round, smooth colonies
Typing:
s.aureus (golden)
Pigments s.epidermidis (white)
s.saprophyticus (lemon)
Antigenic Structure
Staphylococcal protein A(SPA): is the major protein in the cell wall,which is an
important virulence factor because it binds to the Fc portion of IgG.
Teichoic acids
Peptidoglicans
Capsule ( some S. Aureus srain)
II. Pathogenicity And Immunity
Pathogenicity:
coagulase:
staphlolysin and leukocidin:
enterotoxin: heatresistant protein
exfoliation
toxic shock syndrome toxin(TSST)
Diseases:
invasive infection
toxic diseases
food poisoning
toxic shock syndrome
Scalded skin syndrome
Diagnosis of microbiology
Pus swab on blood agar, 37 C, 24 hr
Select colonies for
Gram stain
30

 Plasma coagulase test

Antibiotics sensitivity test
Pus swab blood plate 370C,24hr select colonies

plasma colagulase test

grams stain
antibiotics sensitivity test

Streptococcus
I. Important properties:
Morphology: spherical ,G+ ,arranged in chain
Culture: blood plate: colonies: small, translucent -hemolytic
Antigenic structure: protein antigen/ M antigen; C antigen; P antigen
Classifications :
-hemolytic streptococcus:
-hemolysis: a green zone; opportunistic pathogen
-hemolytic streptococcus:
-hemolysis: a clear zone; pathogen
-streptococcus:
non-hemolytic; non-pathogen
II . Pathogenesis:
Virulence factors:
Streptolysin (streptolysin O; streptolysin S)
Streptococcal pyrogenic exotoxin
Hyaluronidase:
Streptokinase(SK):
Streptodornase(SD)
Diseases: pyogenic diseases; toxigenic diseases; immunologic diseases: rheumatic fever
Diagnosis: Gram-stained smears; culture of swabs; serological test
III.
Pneumococcus (streptococcus pneumoniae)
o Pathogenesis is relate to capsule
o Pneumococcal pneumonia
IV.

Meningococcus
o Pathogenesis is related to endotoxin and capsule
o Meningitis

V.
Gonococcus: Pathogenesis
Neisseria gonorrhoeae (Gonococcus)
I.
Important properties:
Morphology & Staining: resemble paired kidney beans; G-- ; pili
Cultural Characteristics: chocolate agar medium; colonies.
Biochemical reaction: Oxidase test
Antigen: pilus protein; LPS; outer membrane proteins(protein)
Resistance: weakly
II.
Pathogenicity and immunity
1. Important virulence factors: pili; LPSout membrane protein; IgA protease
31

2. Clinical finding:

men:
urethritis
woman: urethritis, cervicitis, cervicocopitis
children: conjunctivis

3.Immunity:
III. Diagnosis of microbiology
1. Grams stain: G- diplococci with PMNs
Culture: Oxidase test
IV. Treatment Personal: hygiene & hospital cross infection
CHAPTER 10: ENTERBACTERIACEAE
Eschericia coli
I.
Important properties:
Morphological and stain : G- rod, flagella, pili
Culture: biological reaction: ferments Lactose; IMViC: + + - Antigen:
O antigen: cell wall; 150; H antigen: flagella; 50; K antigen: capsular; 90
serological types: O:K:H\
II.
Pathogenesis:
Apportunitistic bacilli .
Pathogenic E. Coli
Virulence factors:
Colonization factor (adhesin) : pili; K Ag
Enterotoxin (Exotoxin) :
Heat Labile erterotoxin(LT) : ATP
cAMP
Heat stable enterotoxin(ST) : GTP
cGMP
Endotoxin
*LT:
A subunit A1: ATP
cAMP
outpouring of fluid
A2
B subunit: GM1 receptor
*ST: cGMP

Diseases:
infection outside the enteric tract ; Pyogenic diseases : peritonitis , cholecystitis , pyelitis
diarrhea ETEC; EIEC; EPEC; EHEC; EaggEC
Diagnosis:
o for pathogen: 1. Specimen: urine, stool , blood, Pus, isolation and
identification; Hygienic tests.
Stool swab s-s agar plate pick
Grams Stain
Up The Colonies Biochemical Test
Serological Test
EHEC
Hemorrhagic: Bloody; copious diarrhea; Few leukocytes; Afebrile
Hemolytic-uremic syndrome: Hemolytic anemia; Thrombocytopenia; Kidney failure
Vero toxin/ shiga-like toxin
32

 Inhibits protein synthesis --> intestinal epithelial cells death --> bloody
diarrhea
Invades, holds onto and releases toxins
LT Toxin
Same as cholera toxin structure : 5 B subunits around A
Process
Binds with ganglioside R on cell mem all along I tract
Stim ATP increase [cAMP] stimulate cell
Cell secretes Na+, H2O, Cl-, K+, HCO3- into I tract P increase diarrhea
and Ab pain
Person losses cell nutrition and H2O
III. Treatment and prevention
Shigella (dysentery bacterium)
I.
Important properties:
Morphological and stain : G- rod; nonmotile; non-lactose-fermenting
antigen O antigen: 4 groups ABCD
K antigen
4 species
o S. flexneri, S. boydii, S. sonnei, S. dysenteriae
o According to O Ag
Gen feature
o Nonmotile, non-fermenting lactose, G- rod
o Easily cause drug-resistance strain
Shiga toxin
o Enterotoxic
o Cytotoxic (I protein synthesis, endothelial damage)
o Effected cells die and cause bleeding in epithelium. Blood mixes with feces.
Invades, holds onto and releases toxins
II.
Pathogenesis:
Principal factors:
invasion: pili
endotoxin: feverhypotension shockDIC; bloody diarrhea
enterotoxin (Shige toxin)
Diseases:
Fecal-oral transmission
Dysentery: ferve , abdominal cramps, diarrhea
Bacillary dysentery
Shigellosis: bloody feces, intestinal pain, pus
Immunity: sIgA
Transmission and clinical significance
Mostly young children
Fecal to oral contact
Child to adults
Adult food handlers
4F (flies, fingers, feces, food)
III.
Diagnosis
Specimen: Stool
33

 Quick immuno method/ detect

Immunofluorescent ball test
Coaggultination
Stool swab s-s agar plate pick up
small transparent colorless colonies semisolid agar
double sugar iron agar
* the rapid
* the rapid test

grams stain
biochemical test
serological test

IV.

Principles of prevention and treatment

Prevention
Streptomycin dependent (SD) (Some resistant to)
Dysentery vaccine
Treatment of shigellosis
Manage dehydration
Antibiotics (disease duration diminished)
Salmonella
II.
Important properties:
Morphological and Stain : G- rod; pili; flagellar; non-ferment lactose; produce H2S;
antigen

Antigenic
Type

Location

Properties

Serotype

Antigenicity
Specificity

O antigen

Cell wall

LPS,
Heat stable

42 groups:
A,B,C,D..

Weakly,
Low,IgM
(early)

H antigen

flagella

Protein;
Heat labile

Phase : a,b,c
Phase:1,2,3

Strong, high,IgG
(late,long)

Capsule,
labile

Poly-N-acetyl
D
galactosamineuronic acid

Vi antigen
III.

Weakly
Related virulence
ViAb

Pathogenesis
Principal factors:
Invasion : pili; Vi-antigen
endotoxin:
exotoxin
Virulence Factors: Endotoxins; Capsule; Adhesion; Flagella
Diseases:
Typhoid (enteric fevers)
the organisms enter multiply in the mononuclear phagocytes leads to
bacteremia produce endotoxin fever & other symptom
Enterocolitis:
Septicemia:
34

 Gastroenteritis
IV.
Immunity
V.
Diagnosis: Isolation and indentification of organism:
Specimen
Enteric Fever: blood, marrow, stool, urine
Food poisoning: stool, vomit, food
Septicemia: blood
Widal test:
Concept:
Method:
Tube Dilution Agglutination (widal test)
Salmonella typhii OAg
O>=1: 160 -> active infection
HAg
H>=1:160 -> past infection
S.paratyphi A,B,C ---- HAg
Vi>=1:160 -> carrier
Results: a. normal level of Ab
OAb and HAb
OAb
HAb

Carrier state
VI.
Prevention : 4 Fs (food, finger, flies, feces)
VII. Enteric (Typhoid) Fever --- Bac leave intestine and multiply with cells of
reticuloendothelial sys.
Process

Invades epithelial cell and damages gastroenteritis (acute phase)

Enters LN, spreads through lymph or blood to different organ (Septicemia,

occurs 10-14 days, lasts 7 days)

Enters blood stream again to I tract, damage more severe

Therapy: Antibiotic; Vaccines: Vi Ag
SALMONELLA VS. SHIGELLA

Shigella Gram-negative, non-motile, non-spore forming rod-shaped bacteria

Salmonella rod-shaped, Gram-negative, non-spore forming, motile with flagella
Shigella invades the intestinal epithelial and releases shiga toxins. Salmonella invades the
intestine and then spreads to the bloodstream (bacteremia).
Shigella causes bloody diarrhea, while Salmonella usually does not.
Shigella doesnt ferment lactose. Salmonella also doesnt ferment lactose but produces
H2S.
CHAPTER 11: VIBRIOS

Vibrio Cholerae (Classical biotype and El Tor biotype)

I.
Important properties
Morphological and stain : Short and fine, Curved rod, G, Single flagella, active
motility; Non-spore forming; Oxidase positive; Low resistance to heat, acid, drying;
Readily cultivated, simple nutritional requirements; Aerobic, pH 9 (tolerates alkaline
conditions); Associated with salt water
Culture: Aerobic, PH 9, TCBS agar
Resistance: Lower resistant to heat, acid, drying
35

 Antigenic structures and classification

1). O antigen: total 155, mainly O1 and O139
o O1 and O139 :

Biotypes: Classic, EL tor

Serotypes (Ag factors): Ogawa, Inaba, Hikojima

o non-O1/non-O139
2). Biology:
o Classic biotype: 1817~1923(1st~6th)
o El Tor biotype: in 1961(7th)
o O139 serotype: from 1992 (8th?)
3) Virulence factors of V. Cholerae O1 and O139 Cholera: toxin, single polar
flagella
Pathogenesis and Immunity
Vibrio Cholerae habitat only adherence to the intestinal epithelial cellular surface, not
enter into cells and blood, multiply there and release Cholera toxin and endotoxin.
Pathogenic only for human.
Clinical findings: nausea, vomiting, diarrhea
rice water stool rapid loss fluid and electrolytes

mortality rate: 50% without treatment

<1% with treatment
Immunity: gastric acid; anti-toxin: very strong, reinfection is rare!

II.

Cholera enterotoxin
B Subunit

bind to the receptor of mucous

III.

IV.

A Subunit (A1A2)
A1 through
Cell membrane.
activation
ATP

cAMP

DIARRHEA

Diagnostic Laboratory Test.

Rapid Diagnosis
1). Smear and staining
2). Immobilize test
3). Fluorescin bacteria ball test spa coagglutination: for toxins
Cultures
37
Peptone broth
Smear
PH8-9
68 h
37
agar plate
Colony
Smear identification
PH8-9
10-24 h
Prevention and Treatment
1. Prevention : separate the patients. 4f , water, personal hygiene.
36

V.

Vaccine: its only for 6 months study on oral vaccine.

2. Treatment: supplement fluid, electrolytes; antibiotics: tetracycline
3. Isolate patient; 4F
Examples :
Vibrio Cholerae -- Gastroenteritis
Vibrio parahaemolyticus --- Gastroenterisitis; wound infection; bacteremia
Vibrio vulnificus -- Wound infection; Bacteremia
Cholerae Exotoxin
B subunit: binds to R of mucous
A subunit : (A1,A2)
1. A1 goes through cell membrane
2. Activation with ATP
3. cAMP increases
4. Water and electrolytes secreted from inner cell to intestinal tract
diarrhea, vomiting, stomach pain
CHAPTER 12 : MYCOBACTERIA

Mycobacterium
I.
Important Properties
I.
Morphological and stain: Rod-shaped, aerobic bacteria, acid-fast bacilli(High lipid
content in cell wall); No capsule, no flagellum, no exotoxin and no endotoxin. >50
species.
Mainly pathogen of human:
M. tuberculosis, and bovis- tuberculosis
M. leprae-leprosy
M. aviumintracellulare-opportunistic infection of AIDS
Mycobacterium Tuberculosis
I. Biological properties:
Morphology and stain: slender rod-shaped, G+, Acid- fast stain (Red), L form; Not
classified as either G+ or GAcid-fast staining (Ziehl-Neelsen method)
thick smear
5% carbol-fuchsion (heat, 5 min)
95% alcohol containing 3% HCl (decolor)
methyl blue (1 min, counter staining)
Culture: (Lazygreedy and stubborn) high resistance to heat, difficult to kill.
1). rich nutrients: egg, yolk, potato, glycerol, and complex organic substances
(malachite green), called Lowenstein media
2). obligate aerobes
3). growth rate is much slower: doubling time-18hr, colony-2~4 weeks.
4). grow in clumps or masses.
because of hydrophobic cell surf
permeability of nutrients into cell (reason for slow growth)
Resistances: acids, alkalis, dehydration, drug
37

 Variation: virulence: BCG; drug resistance

II. Pathogenesis
In general: without production of endotoxin or exotoxin; type IV hypersensitivity--important role
1. Constituents of Tubercle bacilli: Lipid, fatty acid and wax are responsible for
delayed hypersensitivity.
2. Pathogenicity
Toxic factor
o capsule(polysaccharides)
o lipids : wax D; sulfatides; phosphatides; mycolic acids; cord factor
o proteins---Ab for diagnosis, tuberculin sensitivity
3. Pathogenesis & Pathology
Two Principal Lesions(pathology)
1). exudative lesions: bacilli, PMN, monocytes
2). productive type: central area, epithelioid cells, fibroblasts, lymphocytes
Spread of Organism in the host: direct extension, lymphatic channel, and
blood stream, bronchi and gastrointestinal tract
Primary infection & Reactivation(secondary) Types
1) Primary infection: usually in childhood, exudative lesion; caseation and
calcifying lymph node; OT test---+
2) Reactivation: usually in adults(bacilli survived in primary lesions). Chronic
tissue lesions; Formation of tubercles; Caseation and fibrosis
4. Immunity and hypersensitivity: old tuberculin (OT), purified protein derivative
(PPD);
Mainly in Cellular immunity: T cell and M,; cytokine
Ab: useful in diagnosis; hypersensitivity
Relationship of Immunity & Hypersensitivity: infections immunity;
hypersensitivity.
5. *Tuberculin Test:
1) material: OT(old tuberculin), or PPD
2) dose: 5 TU/0.1 ml (1~250 TU)
3) reaction: time---48~72 hr.
induration <5mm - never been infected or immunity down
>5mm + infected
>15mm ++ active disease especially in children
6. Diagnostic laboratory test:
o Specimen: sputum, gastric washings, blood, etc.
o Smear Acid-fast stain(104~5/ml)
o Culture: (102~3 /ml)
o DNA detection: <10/ml
o Animal test: Guinea pigs(check lymph nodes)
o Antibodies
o Specimens-sputum, pus, CSF, stool, urine, etc.
o Microscopic examination- Acid fast stain.
o Culture- solid culture media method; slide culture media method.
7. Prevention and treatment:
1. Prevention: BCG inoculation/ vaccination; DNA vaccine
The only available vaccine.
Administered intradermally to children
38

 Creates false positive results in Tuberculin test

2.Treatment: specific chemotherapy; 2 major drugs (first-line drugs(3);
second-line drugs(6))
8. Tuberculosis is easily transmitted because: Air-borne; Highly Resistant; Highly
mutative
CHAPTER 13: ANAEROBIC BACTERIA (Clostridium)
Cl.Tetani ; Cl.Perfringens ; Cl.Botulinum
I.

Common characteristics
Morphology and stain: G+ Bacilli; posses spore.
Culture :
Resistance :
Exotoxin :

II.

Pathogenicity.
Pathogenic factors:
Cl. Tetani

Tetanospasmin.
Tetanolysin;
Diseases:

Cl. Perfringens
Exotoxin (12)
invasive enzyme;

III.

Prevention and treatment.

IV.

Diagnostic laboratory tests.

Cl.Botulinum
Exotoxin (8)

CHAPTER 14: CORYNEBACTERIUM (DIPHTHERIA BACILLI)

Corynebacterium Diphteriae
I.
Biological properties:
Morphology and stain: G+, Club-shaped, Metachromatic granules.
Albert staining (body---blue, metachromatic granules---black)
Albert staining :
Smear Alberts staining solution (5 min) Lugols iodine solution (1 min) check
under microscope
Culture media: Loefflers serum media; potasium tellurite---3 biotypes
Three biotypes of C. diphtheriae :
Properties
Gravis
Intermedius
Mitis
Big,gray
Black in center
Black
Colony size
39

Starch fermentation +
+
Hemolysis
Resistances: not strong; resistance to dehydration; sensitivity to penicillin, chloromphonic
erythromycin
II.
Pathogenesis: Exotoxin produced by Lysogenic Corynebacterium.
In general: spread by droplets or by contact; grow on mucous membrane
1. Pathogenic material:
Diphtheria toxin (important role): produce condition: lysogenic conversion
Fe+2---0.14~0.5ug/ml; structure: as following figure; toxicity: very strong, kill one
cell/one molecule
Structure of Diphtheria toxin
Fragment B: 38 kD, binding
Fragment A: 24kD, toxicity
Action: inhibit polypeptide chain elongation
EF-2 + NAD +
ADPR-EF-2 + NA + H+
EF-2: elongation factor 2
NAD: nictotinamide adenine dinucleotide
ADPR: adenosine diphosphate-ribose
NA: nicotinamide adenine
Susceptible tissue: heart muscle, kidney, liver,
Disease : Endocarditis ; Pseudomembrane ; paralysis of the soft palate.
Immunotoxin:
1. Structure of Diphtheria toxin (DT)

2. DT-390 immunotoxins:

Disease:
1) diphtheria---is an acute respiratory infectous disease
2) illed age
3) the bacilli grow on mucous membrane pseudomembrane toxin blood
distant toxic damage
4) earlier stage---prostration and dyspnea late stage------heart damage
Immunity:
1) specific neutralization anti-toxin by Schick test material and dose:
(subcutaneous injection)
experiment---0.1ml toxin
control side---0.1ml inactivated-toxin
Diagnostic laboratory test:
pseudomembrane or swabs(nose, throat)
Loeffler or tellurite plate
40

Albert staining

virulence test
( Elek plate, ELISA, animal test)

Prevention and treatment:

1. Prevention: active immunization with diphtheria toxoid ; anti-toxin: 1000~3000
Units.
2. Treatment: anti-toxin: as early as possible; sufficient 20,000~10,0000 Units
antibiotics.
CHAPTER 15: ACTINOMYCETES AND NOCARDIOSIS
Actinomycetes
I.
Concepts
normal flora
pathogens
A.israelii
A.bovis
A.naeslundii
II.
Properties of biology
Morphology and staining : filament; actinoid arrangement; G+; prokaryotic;
taxonomic position between fungi and bacteria;sulfur granulesin pus or tissue are
diagnostic

I.

II.

CHAPTER 16: SPIROCHETES

Concept: Being slender, flexible, helically coiled; rapid-motile; Gram-negative,
procaryotic organism; having eight genera; three caused illness (Treponema;
Borrelia; Leptospira)
Leptospiro
1. Properties of biology
1) Morphology : cylinder-like; 0.1~0.2um width, 6~20um length; bent or hooked
ends, like c or s ; G- ; fontana silver-staining- brown
2) Basic structure: outer membrane; axial fibrcles (endoflagellum); protoplasmic
cylinder (cell wall&cell membrane)
3) Culture
media---Korthofs media
culture condition (1~2 weeks)
temperature----28
aerobic
growth results liquid media----translucence
solid media ----dish-like colonies
4) Antigenic structures
surface antigens : glycoprotein, type specific; more than 200 types (49 in china)
inner antigen: LPS, group specific; 25 groups (16 in china)
2. Pathogenicity and immunity
1) Pathogenicity : hemolysin; cytotoxin factors(CTF); endotoxin-like materials
2) Course of disease
Leptospira kidney of animal (pig, mice)contaminated waterhuman
bodyleptoemia
3) Immunity : persistant; humoral immunity
3. Diagnosis and therapy
41

1) diagnosis
observation directly by darkfield microcopy
isolation : Korthofs media; inoculation of guinea pigs
serologic test: microscopic agglutiontest; complement fixation; indirect
agglutination test
2) Therapy: antibiotic; vaccine
CHAPTER 17 : MYCOPLASMA & CELL WALL DEFECTIVE BACTERIA
Concept: Being the smallest; free-living; Procaryotes; their most striking feature is
the absence of a cell wall; highly pleomorphic, can pass through filtres that retain
most bacteria; Colonies minute (<0.2mm), show a fried-egg appearance; Exist in
mouth, respiratory and genito-urinary tracts, common contaminants of tissue-culture.
II.
Properties of biology
Morphology and structure: very small (0.2~0.3um); Wall-less; Multiple
reproductive forms; poly-shapes; Cell member containing a lot of cholesterol ;
Gram--negative; Giemsa staining----light purples; Penicillin--no effect ;
tetracyclin, erythomycineffects
Culture
can grow on artificial media , but have complex
nutritional requirements, including several lipids
grow slowly and required at least 1 week.
form a visible fried-eggcolony
Antigenic structures : GIT , MIT
III.
Pathogenicity and immunity
1. Pathogenicity
often cause surface infections
adhere to a variety of surfaces
toxic materials: neurotoxin, superoxide ions
2. Immunity
cold agglutination reaction :auto-antibody + red cells---- agglutination
IV.
Differences between mycoplasm and bacterial L form
MYCOPLASMA
L FORM
+
Heredity relation to bacteria
+
Restored to bacteria
Slow, small
Fast, large
Growth on solid media
Light turbid
Very turbid
Growth in liquid media
+
Requirement for cholesterol
VI.
Mycoplasma pnemonia
1. a short filament, 2 to 5um in length
2. Pathogenicity
transmission--------aerosol droplets
disease-----primary atypical pneumonia
3. Diagnosis: Isolation and cultures; Serologic test; PCR or ELISA
VII. Others mycoplasma : Ureaplasma urealyticum---- nongonococcal urethritis
M. pneumoniae --primary atypical pneumonia; M. hominis; genito-urinary
infections
CHAPTER 18: CHLAMYDIA
I.

I.

Concept
42

II.

III.

Being a family of obligate intracellular parasites

have a unique development cycle
can pass through the filters.
Characteristics:
G-, spheric or ellipsoidal: Cell wall without peptidoglycan
Development cycle: elementary body reticular body
Both DNA and RNA
Having ribosome, energy parasite;
Sensitivity to some antibiotics
Properties Of Biology
1. Development cycle and staining

2. Culture --- living cells

3. Classification
antigens
genus specific----LPS
species specific---MOMP
type specific---MOMP

IV.

V.

I.

4. Resistances
Pathogenicity and immunity
1. Pathogenicity: unknown; inhibition of host cell metabolism; toxic materials; IV
allergy; inflammation
2. Immunity: both humoral and cellular immunity; weak, short time
Diagnosis and treatment
1. Diagnosis: staining inclusion body; IFA; PCR culture
2. Treatment: rifampin etc
CHAPTER 20: RICKETTSIA
Concept
Procaryotic organism that are obligate intracellular parasites
Obligate intracellular parasites.
43

II.

III.

IV.

V.

Most of them infect both human & animals, transmitted by arthropod vectors.
important species:
Share common antigens with certain strains of proteus (ox19, ox2, oxk).
Weil-felix test is a famous test to help diagnosis
Important rickettsia Characteristics
Arthropod-rodent and vertebrate reservior
Related to arthropods closely
Size between bacteria and virus
Pleomorphic
Intracellar multplication
Sensitivity to antibiotics
Contain DNA and RNA
Properties of biology
Morphology: pleomorphic (rod-shaped to coccoid); Gram negative; Giemsa
purple or blue
Culture: require living cells
Antigenic structures

group specific antigen: lime layer

type specific antigen: cell wall

cross-reaction antigen: Weil-Felix reaction

Pathogenicity and immunity
1. Pathogenicity
materials : endotoxin; phospholipase A
pathology: perivascular infiltration; twice rickettsemia
manifestation: headache, fever, rash
disease: epidemic typhus; endemic typhus; scrub typhus; Q fever
2. Immunity: cellular immunity
Diagnosis and treatment
1. Diagnosis
isolation culture --- susceptible animal
serologic test --- Weil-felix reaction
2. Treatment : tetracyclin and chloramphenicol etc.

CHAPTER 21: VIRUSES ASSOCIATED WITH RESPIRATORY INFECTIONS .

INFLUENZA VIRUS
I.

II.

Categories
o Orthomyxovirus: influenza virus.
o Paramyxovirus: parainfluenza virus; mumps virus; measles virus; respiratory
syncytial virus.
o Adenovirus: human adenovirus.
o Miscellaneous: rubella virus; rhinovirus; coronavirus.
Characteristics of respiratory virus infection
1). High incidence, quick transmission and serious harm.
2). Not only causes the respiratory diseases, but also causes the diseases in the other
parts of the body.
INFLUENZA VIRUS
44

I.

Biological properties
Morphology and sructure
1). Morphology: spherical, 80-120 nm,enveloped.
2). Structure:
(1) Internal layer: nucleocapsid.
a. Viral nucleic acid: -ssRNA, 7~8 segments
b. Viral capsid: nucleoprotein(NP)+RNA RNP
(2) Middle layer: Matrix protein(M1 and M2).
(3) External layer: envelope.
a. Hemagglutinin (HA): trimer, stalk
structure:
566 aa endoplasmic reticulum removed
signal sequence cleaved into HA1 and HA2
function:
HA1: binding to receptor; HA2: membrane fusion
HA can agglutinate with RBC if human, chicken.
b. Neuraminidase (NA): tetramer, mushroom facilitates release of mature
virion
Type specific Ag: RNP & M
influenzavirus A, B, and C
influenzavirus A, B---human and animal A
human strains of type B
influenzavirus C--- human and swine C
Subtype specific Ag: HA & NA (5:1)
H1~15
N1~9
ORTHOMYXOVIRUS
type A, B, C : NP, M1 protein
sub-types: HA or NA protein

PARAMYXOVIRUS

45

II.

III.

IV.

Antigenicity and typing

There are 2 kinds of antigens:
1) type specific antigens: nucleoprotein(NP) and M protein
2) subtype specific antigens: HA, NA (surface glycoproteins)
3) Relationship between the antigenic variation and epidemiology
(1) Epidemic condition
a. Antigenic drift: no new subtype appearance
epidemic--- small or median
b. Antigenic shift: new subtype appearance
epidemic--- large scale
(2)Variation of HA and NA of influenza virus
a. Antigenic drift: mutation range---smaller, <1%
b. Antigenic shift: mutation range---larger, >20%~50%
(3) Reasons
a. RNA consists of 8 segments.
b. These RNA segments can reassortment freely.
c. The variation of the HA and NA10-3~10-5
Where do new HA and NA come from?

15 types HA

9 types NA: all circulate in birds

Pigs: avian and human

Influenza virus culture
1) embryonated eggs
2) primary monkey kidney cells
3) life cycle as follows: 8~10 hours/cycle
Resistance: not strong, especially for lactic acid.

46

V. Pathogenicity and immunity

1) Source of infection: patients and carriers.
2) Transmission: aerosol
3) Susceptibility: in general
4) Seasons: winter and spring
5) Incubation time: 18~72 hours
PATHOGENESIS
patients or carriers

viruses
respiratory tract
release toxoid substance
invasive
into
neighbouring cells
blood

respiratory acute inflammation

toxic symptoms
(lighter)
(heavier)
Transmission
Aerosol: 100,000 To 1,000,000 per Droplet
Incubation: 18-72 Hr
Shedding
Immunity: SIgA, IgG, IgM .
a. Transient immunity, which lasted within 1-2 years and SIgA is more important.
b. Antibodies to HA, NA, share the protect functions.
VI. Laboratory diagnosis
1. Isolation and identification of viruses:
isolation---first generation grow slowly
identification--- hemagglutination; hemagglutination inhibition (type and subtype)
2. Serological diagnosis:
antibody to HA; NA and RNP.
assay: hemagglutination inhibition
VII. Prophylaxis and treatment:
47

1. Public and personal hygiene: avoid gathering; evaporate lactic acid

2. Vaccination:
inactivated virus---from 1941, USA
sub-unit vaccine---from 1980, UK
3. Treatment: Amantadine hydrochloride; Antibiotics (prevention complication)
CORONAVIRUSES
Genome:
5 polymerase---S---E---M---N---3
ORF1a/1b: RNA polymerase(Rep)
S: virus attachment and fusion
E: virus assemble
M: stabilizing core
N: equal to nucleoprotein of influenza
SARS (severe acute respiratory syndrome)
CHAPTER 22: HEPATITIS VIRUSES
In general,heptatitis viruses are composed of HAV,HBV,HDV,HCV, HEV and HGV .

I.

Hepatitis A virus (HAV)

Biological Properties
Morphology and structure: Typical enterovirus in piornavirus family
27nm; sphere;
nonenveloped icosahedral nucleocapsid
having a single-stranded RNA genome
HAV protein having antigenity, can cause
neutralizing Ab

48

II.

1.

2.
3.

Culture
replicates in the cytoplasm of the cell
Animal inoculation and cell culture
Resistance : weak; stable under 560C for 30 weeks
Pathogenicity and immunity
1. Infection sources --- acute patients
2. Transmission pathway --- fecal -oral route mainly
3. Pathogenesis --- direct damage and immunopathogensis
4. Immunity --- persistant immunity after recovery
Laboratory diagnosis
detection of IgM antibody is the most important
A 4-fold rise in IgG antibody titer can be used
Isolation of the virus in cell culture
Treatment & prevention
No antiviral therapy is available
Inactivated HAV is used to active immunize
Passive immunization with immune serum globulin
Hepatitis B Viruses
---1964, Blumberg (Australian)HBsAg
---1970, Dane (England)---- HBV viral particle
Morphology and structure
Dane particle
42nm, double shells, contain all of antigen) (HBsAg,
preS1, preS2 HBcAg, HBeAg) and core (DNA, DNA
polymerase), infectivity.
Spheric particle-- 22nm, containing HBsAg
Pipe-like particle-- 22nm, long 50-700nm
Genome structure : 3.2bp in length; consisting doublestranded circular DNA; the DNA polymerase has both reverse transcriptase and DNA
dependent
1) S-strand
2) L-strand--- 4 ORF
S----HBsAg, preS1, preS2
C----HBcAg, HBeAg
P----DNA polymerase
X----HBxAg
4) replication cycle:
5)
49

infectious virion attaches to cells

uncoating

partially double-stranded covered to covalently closed circular double-stranded

2.1KbmRNA
3.5KbmRNA

protein of outer
protein of inner capsid as temple for replication of
HBV-DNA

Capsid
a negative strand DNA copy

a positive strand DNA copy

Antigenic Components And Antibody
1) HBsAg and HBsAb
4) HBcAg and HBcAb
HBsAg:
HBcAg:
subtype: adr, adw, ayr, ayw
183 aa, Mw, 22000
location:surface antigen
Antigen associated with core of HBV
symbol of infection
cannt detected in blood
HBsAb: neutralizing, a protected
HBcAb:
antibody
positive during window phase
IgM HBcAb is an indicator of recent
2) Pre-S2 and Pre-S2-Ab
disease
Pre-S2:
5) HBeAg and HBeAg -Ab
55aa linked to amino terminal of
HBsAg (middle molecule)
HBeAg:
mediated virual adsorption by PHSA-R
Mw, 18 KD ; subtype - e1, e2, e3
A second , different antigen in HBV
Pre-S2-Ab: a protected antibody
core
3) Pre-S1 and Pre-S1-Ab
But is a soluble antigen
Pre-S1:
Being a indicator of transmissibility
119aa linked to amino terminal of
HBeAb: indicates low transmissibility
pre-S2 (large molecule)
Pre-S1-Ab:
Animal model and tissue culture
susceptible animal
chimpanze, duck
tissue culture-----hepatoma cell transinfected by HBV gene
Pathogenicity and immunity
1. Infection sources : HB patients; chronic carriers of HBsAg
2. Transmission pathway : blood or blood product; sexual intercourse; mother to
newborn
3. Direct viral-induced cytopathogenicity
4. Immunopathology: Cytotoxic T cells mediate an immune attack against the viral
antigen , and inflammation and necrosis occur.
Cellular immutity
normal------acute hepatitis
strong------- fulminant hepatitis
50

weak------- chronic
Immunocomplexes
extrahepatic manifestations
arthritis, nephritis, dermatitis etc
Self-immunorresponse
LSP TC or TD damage of infected cell
5. Hepatitis B characteristics: Extrahepatic manifestation; Chronic forms; Long
time carrier state; Cirrhosis
6. Immunity: HBsAb, pre-S2Ab having protective; Cellular immunity can eliminate
virus in cell
Diagnosis
1. Specific Antigen and antibody
Method: ELISA and RIA
Immunodiffusion ( for subtype )
West-blot ( for peptide fragment )
Significance: HBsAg persisting in blood for at least 6 months; Indicate
chronic carrier
Window phase: HBsAg has disappeared but HBsAb is not yet detectable.

2. HBV DNA: DNA hybridization techniques; Spot and Southern; PCR

3. Dane particle: IEM
4. DNA polymerase
Principle of prevention and treatment
1. Treatment of patients
rest ; nutrition
Medicine: IFN; chinese medine;
2. Cut off the infectious route : Isolation of patients with acute hepatitis; Screen the
blood donors; Sterilization of instruments
3. Immunization
1) Active immunization
Vaccine from plasma of HBsAg of symptomatic carriers; genetic engineering;
artificial polypeptide of synthesis
Procedure--- usually for newborn infants0-1-6
2) Passive immunization: injection HBIg; new infants born from mother whose
51

HBsAg positive; individual exposed to HBsAg-positive blood

CHAPTER 23: HEMORRHAGIC FEVER VIRUSES (HEMORRHAGIC FEVER WITH
RENAL SYNDROM VIRUS)
Biological properties:
Sphere, diameter=100nm, SSRNA; Envelop; Stable: in 4-20
Animal reservoir: rodents
Transmission and pathogenisis:
Incubation period was 4-16 days.
Bleeding shock, renal failure etc.; Locality and season.
Microbiological diagnosis
1. Viral isolation: from acute serum. Pulmonary. kidney. etc. grows in cultures of vero-E cells
or A549 cells
2. IgM or IgG Ab:
CHAPTER 24: RETROVIRUSES
Basic Properties:
1. +ss RNA, but double strands, three structure genes
2. Reverse transcriptase (RT)
3. Spherical, 100 nm or so
4. Replication:
RT
RNaseH/DDDP
Insertion
RNA+ cDNA(RNA:DNA hybrid)

ds DNA

host DNA

provirus transcription

mRNA
RNA protein assemble mature viruses

assemble mature virus

Classification:
Three Subfamilies totally
1. RNA Oncovirinae: causing leukemia and tumors, and Human T cell Lymphotropic Virus--HTLV- I, II, V types
2. Spumavirinae:
3. Lentivirinae: Human Immuno deficiency Virus (HIV)
HIV
Epidemiology: 15000/day=1/6sec
Transmission :
1). infectious Sources:
AIDS Patients; HIV Carriers;
2). infectious routes:
(1) Sexual transmission:
(2) Infectious blood: Blood products, Syringes
(drug abusers), Transplantation, etc.
(3) Mother-baby born:
General properties : locality; young people(male>Female); homosexual
HIV biological properties:
52

1. Genome: +ss RNA, 9 genes, 9.7kb

Structure genes:
po1enzymes(P66/P51, proteinase)
gagCapsid(P24, P17, P7)
envenvelope (gP41, gP120, gp160)
Regulatory genes: vif, vpu, vpr, tat, rev, nef;
2. Replication: Reverse transcription
3. Variation: relative higher, especially in gP120 gene subtypes of HIV-1: 11
subtypes
4. Resistance: weak; 56oC for 30 minutes room temperature for 7 days
Clinical Transmission
Clinical symptoms:
long incubation; 100% died;
HIV
CD4 receptor CCR5, CXCR4
T4 , Mononucleus cell, M

Cell death
T4/T8 <1

Cellular Immunity
HIV transmission to central Nervous system, other monocyteMNK
Pathogenic mechanism and clinical symptoms:
CD4 recepter;CCR5;CXCR4 ; T4/T8 <1; Cellular Immunity
ARC: feverweightfatiquediarrheaKaposi Sarcoma
Opportunistic infection: Fungi. Bacteria..
Diagnosis:
1. Anti-HIV (primary test and confirming test) IF+ELISA; Western blot+RIA.
2. T4, TH/TS <1
3. Isolation of HIV from blood.
4. HIV RNA : PCR.
Treatment: Medicine: AZT. DDI; Vaccine: GP160; Gene therapy:
Control: health education; blood; blood products; drug abuser; sexual; mother-baby
born
Anti HIV drugs : Nucleoside Reverse Transcriptase Inhibitors (NRTI); NonNucleoside Reverse Transcriptase Inhibitors (NNRTI); Protease Inhibitors (PIs).
HERPESVIRUS
I.
INTRODUCTION
Biological properties:
1. ds DNA: Surrounded by protein coat (Nucleo-capsid) icosahedral symmetry.
2. Envelope: (Size 150-220 nm): Spherical virion
3. DNA replication in cellular nucleus, and budding from nuclear membrane
4. produced intranuclear acidophilic inclusion body.
II.
CLASSIFICATION
Microbiolocal diagnosis
53

1.Isolation of virus:(cell culture); 2.DNA Hybridization; 3..Antibody: IF, ELISA

CHAPTER 25: PRION
Basic Properties:
Prions are infectious proteins that differ from all other known pathgons and are belived to be
devoid of nucleic acid. Prions are highly resistance to conventional inactivation procedures.
Pathogenic mechanism and diseases: Prion diseases are neurodegenerative diseases, which
affect humans and a variety of domestic and wild animal species.
Small; Filterable; Need host cells; No machinery for energy generation of protein synthesis
Different from viruses

No dectectable virions in infected tissues

No detectable virions in purified infectious material

If nucleic acid is present, very small

Very resistant
Resistant to or only partially inactivated
Inactivated by
1. Formaldehyde
1. Autoclaving
2. Ethanol
2. 5% sodium hypochlorite
3.Sodium hydroxide
3. Glutaraldehyde(
4. urea, other protein denaturants
4. Ultraviolet and ionizing irradiation
5. Proteases
6. Non-ionic detergents

Structure
A prion is composed of proteins encoded by a normal cellular gene.
The protein, designated PrPc, is converted from a normal benign form into a diseasecausing form by a change in conformation to a protein designated PrPsc (for the
scrapie protein).
Brain extracts from scrapie-infected
animals contain PrPsc, which is not found
in the brains of normal animals.
PrPsc is the prion that is responsible for
transmission and infection.
The conformational change is also the
way that prions multiply
Contact
with
PrPsc
results
in
conformational change of the normal host
cell protein PrPc and the formation of
additional PrPsc.

How can this model explain the sporadic, acquired or inherited form of the disease?

54

During scrapie infection, prion protein may aggregate into birefringent rods and form
filamentous structures termed scrapie-associated fibrils (Fig 441), which are found in
membranes of scrapie-infected brain tissues.

Pathogenesis

Kuru

CJD(Creutzfeldt Jakob disease)

GSS(Gerstmann Straussler-Scheinker syndrome)

FFI(Fetal familial insomnia)

Scrapie(sheep and goats)

Bovine spongiform encephalopathy

Transmissible mink encephalopathy

Chronic wasting disease of mule deer

CHAPTER 26: GENERAL PROPERTIES OF FUNGI
Characteristics:
1. The largest one among all the microorganism.
2. Eucaryotic, including
unicellularyeasts.yeast-like organism, cryptococcus neoformans
Multicellularmolds; hyphae spores
3. Easily to cultivate, low nutrient; PH (4.0-6.0); temp (22-28); high humidity; plenty of O2;
rich sugar
Sabourauds medium
Slide culturefavorable to observe the shape of molds
Colonies: three types
4. Resistant to dryness: alcohol; common antibiotic. Susceptible to Bromogeramine()
5.A great number of species(>105), widespread, participate in the cycle of nature, useful foodindustry ,contaminate cultures. food and drink crops etc. Only a small part are pathogenic.
Classification and disease
1. Dermatophytes, causing superficial mycoses (skin, hair, nail); Causing
opportunistic mycoses
(1) Candida albicans: Chlamydespores; Pseudohypea yeast-like colonies
Causing candidiasisoral (Cutaneous; Alimentary; Vaginal; Systemic)
(2) Cryptococcus neoformans: very thick capsule; causing cryptococcosis
3. Causing allergy
4. Causing Mycocoxicosis
5. Causing Cancer-related disease
55

 Detection:
1. direct observed by microscope: hair
2. Culture:
3. Animal inoculation: cryptococcus neoformans.
Major Pathogenic Fungi: Candida albicans; Cryptococcus Neoformans
Histoplasma capsulatum;Penicillium marneffei .

56