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INTRODUCTION  Pasteurization – destroy

pathogens in milk by heating the


History / Milestones of Microbiology: milk at 63 degrees Celsius for
 Old testament 30 minutes or 72 degree Celsius
o Leprosy – 1st disease discovered for 15 seconds
o Book of Leviticus – 1st recording of laws o 1865
concerning public health  Joseph Lister
 personal hygiene • Father of Antiseptic
 burying of waste materials • used phenol as spray in
 isolation of the sick operating rooms to
 13 century
th reduce infections
o Roger Bacon • the 1st to use culture
 invisible creatures as pathogens media to isolate
 15 century
th bacteria into pure
o 1546 – Giralamo Fracastoro / Girolamo culture
o 1870
Fracastorius
 early observations on the nature  Germ Theory of Disease
and spread of infectious disease • Specific disease is
 father of scientific epidemiology caused by a specific
type or microorganism
 most famous work was the
narrative poem  Robert Koch
Syphilis sive morbus Gallicus • proposed the Koch’s
• described a new postulate :
sexually transmitted o postulate was
disease, now called formulated to
syphilis, and its provide proof
treatment that a specific
 16th century bacterium
causes the
o 1667 – Anton van Leeuwenhoek / disease
Antony van Leeuwenhoek
o postulate must
 father of Microbiology be fulfilled for
 invented the simple microscope the micro-
– one lens microscope organism to be
 demonstrated the presence of considered as
invisible organisms (microbes) the etiologic
 he called the microbes as agent (the
“animalcules” cause) of the
o 1678 disease
 Robert Hooke  Koch’s Postulate
a.
 invented the compound
Organism must always be
microscope
found in the diseased
 1800 – 1900’s : animal but not in healthy
o most of the bacteria were identified one
o “Golden era of Bacteriology” b.
o 1850 Organism must be isolated
 John Tyndall from the diseased animal
• he found out that and grown in pure culture
bacteria with c.
endospores were The organism isolated from
resistant pure culture must reproduce
• tyndallization – the disease when
endospores destroyed inoculated into a susceptible
by boiling and cooling animal
three (3) times d.
Organism must be re-
isolated from the
Introduction – Francis Anthony V Marfori MD, BSN 2
Yvonne Agnes Villarta-Marfori MD, BSN

experimentally-infected o developed the technique of vaccination


animal by attenuated (chemically weakened)
o 1892 virus
 Dimitri Ivanofsky / Iwanosky  Elie Metchnikoff
• Virus called “liquid o proposed the Cellular Theory of
poison” Immunity
• tobacco virus : 1st virus  Jules Jean Baptiste Vincent Bordet
discovered; tobacco o proposed the Humoral Theory of
mosaic disease was Immunity
traced to the
 19th century
suspension that was
o 1947 – disinfection of hands with
passed thru bacterial
solution of chlorinated lime
filters
o Florence Nightingale
o 1898
 developed modern principle of
 Martinus Beijerinck / Bujenick
nursing
• discovered that virus is
 organization of hospitals to
an obligate parasite
reduce spread of disease
 1928
o Penicillin was discovered by British
bacteriologist Sir Alexander Fleming Molecular Biology:
 1940  In 1953, American biochemist James Watson
o most of the viruses were discovered and British biophysicist Francis Crick proposed
with the advent of electron microscope the now-famous double-helix model of DNA
(E/M)
 the study of DNA composition / sequence of an
o rapid advances in virology with the
organism
invention of the E/M
Microbiology
Concepts of Immunity:
 ancient China  the study of living organisms too small to be
o variolation seen by human eyes (microbes)
 Major Groups of Microbes :
 type of vaccination against small
Bacteria : Bacteriology
pox
Viruses : Virology
 healthy people inhaled a powder
Fungi : Mycology
made from scabs of healing
Parasites : Parasitology
pustules of small pox
a. Protozoa – single-celled
 Edward Jenner b. Metazoa – multicellular, called
o Father of Vaccination helminths (worms)
o 1st vaccine (“vacca”) : cowpox vaccine to
prevent smallpox in man  Classification :
o 1796 inoculated an eight-year-old boy Bacteria : prokaryote kingdom
with cowpox virus; six weeks after the Fungi
boy's reaction Jenner reinoculated him Protozoa : protists kingdom
with smallpox virus, finding the result Helminths : animal kingdom
negative Viruses are not cells but can replicate
 Ignaz Semmelweis only within cells
o late 1800’s
o found out that puerperal sepsis caused Major Groups: (Microbes)
by hands of doctors and midwives
 Louis Pasteur Prokaryotes Eukaryotes
o French chemist/ bacteriologist Eu = true ; karyon =
o founded the science of microbiology nucleus
 simple  organisms
o coined “vaccination” from the word
organisms whose cells are
“vacca” the first vaccine composed of more complex
Introduction – Francis Anthony V Marfori MD, BSN 3
Yvonne Agnes Villarta-Marfori MD, BSN

single cells  well-defined  reproduce by binary fission : referred to


 lack a nucleus as exponential or logarithmic growth
well-defined  well-defined o 1 bacterium produce 16 bacteria after 4
nucleus nuclear membrane generations
 nucleoid o the doubling (generation) time ranges
consists of a
from 20 min (E. coli) to >24 hrs (Myco.
single circular
TB)
molecule of
 motility is by flagellar-like structure
loosely organized
DNA  most are enclosed in a rigid wall made up of
peptidoglycan
 lacking a
nuclear memb  photosynthesis, if carried out, is anaerobic
and mitotic  rickettsiae, chlamydiae and mycoplasma –
apparatus referred to as “small bacteria”
 no  organelles  rules in writing names of bacteria
intracellular in the cytoplasm o genum and species names are written
organelles o 1st letter in genum is capitalized; 1st letter
 ribosome in species not capitalized
s are free in the  chlorophyll
cytoplasm is contained within
o both names are italic or underlined
 chlorophy a chloroplast o if underlined, one line for genum and
ll is dissolved in another line for species name
the cytoplasm o eg:
 only 1  multiple  Neisseria gonorrhea
chromosome chromosomes  Salmonella typi
 most  10 characteristics – these characteristics are
have a rigid used for identifying and classifying bacteria
external wall that o I morphology
contain o II staining
peptidoglycan
 cell
o III motility
memb. has no o IV growth
sterols except the o V atmospheric requirements
wall-less o VI nutritional requirements
Mycoplasma o VII biochemical / metabolic
 multiply  reproductio activities
by binary fission n is by mitosis o VIII pathogenicity
 motility  most o IX amino acid sequencing of
occasionally protozoa are motile
proteins
present (moves by cilia,
pseudopod or o X genetic composition
flagella) but fungi
and virus are
nonmotile
 eg:  eg: I Morphology
bacteria protozoa fungi Major Classes of bacteria
chlamydiae
rickettsiae
1. 3 basic groups based on shape : (determined by
its rigid cell wall)
mycoplasma
cocci : sphere-shape
blue-green bacilli : rods
algae spirochetes : spiral

o Bacterial arrangements are determined


BACTERIA
by the orientation and degree of
 unicellular
attachment of the bacteria at the time of
cell division
Introduction – Francis Anthony V Marfori MD, BSN 4
Yvonne Agnes Villarta-Marfori MD, BSN

o Cocci : 2. Isolatio
 clusters (grapelike) n of organism in pure culture
: staphylococcus 3. Bioche
mical tests
 in pairs 4. Animal
: diplococcus inoculation
 in chain 5. Immun
: streptococcus ologic reaction
 in groups of 4
: tetrads Microscope :
 in groups of 8 1. Light microscope (L/M) (Bright-field) :
: sarcina o a 2-lens system :
o Bacilli :  objective lens (lens nearest the
 Diplobacilli : object)
• Snapping (V-shape)  ocular lens (lens nearest the
• Slipping (parallel ) eye)
 Coccobacilli o organism looks dark; background is light
: short, plump rods
 Fusobacteria 2. Dark-field microscopy :
: long, tapering o dark background and object is
illuminated
 Sporing bacilli o organism is living, unstained
: with spore
o eg: Treponema pallidum
 Streptobacilli
: in chain 3. Phase-contrast microscopy :
 Bacillus with squared ends o alive and without staining
: B. anthracis o internal details can be seen
 Bacillus with rounded ends o contain special filters, diaphragm, which
: salmonella splits the light beam separated light
 Curved rods passes around and through the object
: vibrio cholera o small differences in the densities of the
o Spirals : object shows up as different degrees of
 Spirillum brightness and contrast
: rigid
4. Flourescent microscopy :
 Spirochete
o organism is coated with fluorescent dye
: flexible

2. Intracellular obligate parasite : Rickettsiae illuminate with UV light


o As electrons in the dye are excited, they
3. Cell wall-less : Mycoplasma move to a higher energy level, then
quickly drop back giving off the excess
energy as visible light object fluoresce
Sizes largest bacterium : bacillus anthracis 5. Electron microscopy :
(2µ ) o resolving power is 300X that of L/M
smallest bacterium : chlamydia and o for viruses
rickettsia
smallest virus picorna virus
30µ (0.001 inch)
: smallest scope of the eye II Staining
Staining Techniques:
Methods of Identification of Bacteria 1. Simple
1. Use of staining
microscope (stained or unstained)
Introduction – Francis Anthony V Marfori MD, BSN 5
Yvonne Agnes Villarta-Marfori MD, BSN

o small amount of bacteria is placed in a  All bacilli are gm (-) except :


droplet of H2O on a glass slide sporeformers (clostridium,
o air-dry bacillus), C. diphtheriae,
o heat-fix (slide passed thru a flame) : Mycobacteria and Nocardia
 binds bacteria to the slide  All spirochetes are gm (-)
 kills most of the bacteria o Significance :
 prepares the bacteria for  Gm (+) bacteria are :
staining • more resistant to
o slide flooded with basic dye (crystal oxidizing agents, alkali,
violet or methylene blue) for 1 minute proteolytic enzymes
o positively charged dye attracted to • more susceptible to
negatively charged bacterial cytoplasm acid, detergents, sulfa,
antibiotics like penicillin
2. Negative staining G
o bacteria on glass slide
o do not heat-fix 4. Acid-fast staining:
o fix smear by heat
o add acid dye (congo red or
o cover with carbolfuchsin
nigrosin)mix air-dry
o dye which is negatively charged is o wash with water
repelled by negatively charged bacteria o decolorize with acid alcohol (95%
o stain gathers around the cell ethanol containing 3% HCl)
o wash with water
3. Gram staining o counterstain with Loeffler’s methylene
o 1884 – Dr Hans Gram blue
o differential staining technique o Acid-fast bacteria :
o Steps :  once stained, bacteria not easily
 air-dry and heat-fix the smear decolorized with acid alcohol;
 stain with crystal violet acid fastness – due to mycolic
acid (resist decolorization)
 stain with Gram’s iodine solution o Result :
to form crystal violet – iodine
complex (blue)  Acid-fast organisms : pink or
red
 decolorize the smear with 95%
alcohol – what is decolorized is  Non acid-fast : blue
the iodine complex o Acid-fast organisms :
 counter stain with safranin  Mycobacteria
• gram (+) – not  Nocardia
decolorized; remains
blue because of a III Motility
thicker cell wall and the  motility is associated with the presence of
gram’s-iodine complex flagella or axial filaments
is well embedded in the  general rule
wall o spiral shaped and half of the bacilli –
• gram (-) – decolorized motile
to red; have high lipid o cocci – generally non-motile
content in its cell wall
which is dissolved by IV Growth
alcohol  leakage of  an increase in the number of organisms rather
crystal violet-iodine than in their size
complex  in the laboratory, we grow bacteria in a culture
medium

o General Rule :
 All cocci are gm (+) except :
Neisseria  culture medium
Introduction – Francis Anthony V Marfori MD, BSN 6
Yvonne Agnes Villarta-Marfori MD, BSN

o area where microorganisms are the type of gm


controlled (-)
o Nutrient agar o phenylethyl
 is the standard bacteriological alcohol (PEA) –
medium grows gm (+)
 agar serves as solidifying agent o colistin-nalidixic
 composed of : acid (CAN) –
• H2O grows gm (+)
• peptone(protein from
o Thayer-Martin
soybean) agar (chocolate
• agar agar + nutrients
+ antimicrobial)
• dried beef extract
– favors growth
o Nutrient broth
of Neisseria
 a liquid medium gonorrhea
 agar is omitted o mannitol salt
agar (MSA) –
Culture: for salt tolerant
 Classify culture medium in terms of: bacteria
1. Physical state: o sabouraud
Solid = contains at least 2% dextrose agar –
agar for fungi
Semisolid = 0.5-1% agar  enrichment media
Liquid = no agar
• allows prolific growth for
certain bacteria
2. According to composition: • supplied with growth
a. factors
synthetic and non-synthetic like albumin, • broth or solid medium
meat containing a rich supply
b. of special nutrients
selectivity and enrichment (growth factor) that
 non selective (simple) promote growth of
• no growth factors fastidious organism
• any ordinary bacteria • eg: chocolate agar –
can grow in it nutrient agar +
 selective media powdered hemoglobin
• with dyes that inhibit  differential media
growth of certain • with use of dyes, it
bacteria and allow determines the type of
growth of other kind growth
bacteria • eg: eosin methylene
• added inhibitors that blue (EMB) agar
discourage growth of
certain organism  Specimen for culture :
• eg: o Blood
o blood agar – o Spinal fluid pure bacterial
favor growth of o Closed abscess specimen
gm (+) cocci
o MacConkey
agar – inhibits streak over culture medium
growth of gm incubate
(+); also a
differential o stool
media
o sputum mixed
(differentiate
organisms colonies
Introduction – Francis Anthony V Marfori MD, BSN 7
Yvonne Agnes Villarta-Marfori MD, BSN

o body orifice • Superoxide dismutase


o needed to
Bacterial Identification based on detoxify
1. Colonia superoxide
l morphology : radical
o due to bacte. motility or post-fission • Catalase/ peroxidase
bacteria movement o needed to
a. size convert H2O2 
b. shape : convex, flat, umbilical H2O
center  Subtypes
c. color • Obligate
d. smell o vulnerable to
e. edges : smooth, wavy, even traces of
serrated O2
f. adherence to culture media o requires
(serologic properties) anaerobic
f.1 Mucoid (M) : environment
 characteristic of o eg: Clostridium
organisms producing slime tetani
or capsule • Aerotolerant
 virulent bacteria o organism
 eg ; streptococci, K. possess
pneumoniae and H. superoxide
influenzae dismutase but
no catalase or
f.2 Smooth (S) peroxidase
 characteristic of freshly o does not
isolated organism require oxygen
 mild type virulence but survive in
 eg; E. coli, salmonella, atmospheres
shigella containing
molecular
f.3 Rough (R) oxygen
 produced by mutant
strains without surface • Facultative anaerobes
protein o capable of
growth both in
 generally avirulent an environ of
except :B. anthracis and
free O2 or no O2
Myco. Tb
o from 0 oxygen
to 20-21%
2. Microscopic
oxygen
morphology and staining reaction
o eg: Escherichia
V Atmospheric Requirements coli
 classify bacteria on the basis of
their relationship to oxygen and carbon dioxide
 Types of organisms based on atmospheric 2. Aerobes
requirements :
1. Anaero  need O2 for respiration
bes  Subtypes :
 organisms that cannot grow in • Obligate aerobes
the presence of free O2 o cannot grow
with no O2
 Why can’t anerobes grow in the o atmosphere
presence of O2?
containing
Because anerobes do not have:
Introduction – Francis Anthony V Marfori MD, BSN 8
Yvonne Agnes Villarta-Marfori MD, BSN

molecular  Organic matter : complex


oxygen polysaccharide
comparable to o Inorganic matter :
that found in  S N2 NH3 nitrate CO2
room air (20-
21% oxygen)
o Patterns of Nutrition :
o eg:
 Autotrophs (lithotroph)
1. Nocardi
a sp • group of bacteria which
2. Psudo synthesize their
monas nutrients from simple
sp sources (CO2, H2O)
3. Mycoba using solar or chemical
cterium energy
tubercu  Heterotrophs (organotroph)
losis – • uses energy-rich
predilec organic carbon
tion for molecules (glucose)
the from the environment
apices • Saprobes
of the o those that feed
lungs exclusively on
4. Bacilus dead organic
sp matter like
• Microphilic compost
o grows best at • Parasite
low O2 tension o feed on living
like in the GIT, matter
GUT
o about 5%  energy source
oxygen o sun
o eg: o oxidation process from chemical
Campylobacter compounds
sp o suitable environment

3. Capnophiles / capnophilic organisms o Type of organisms based on energy


 grow better in the presence of source :
increased concentration of  photosynthetic – energy source
carbon dioxide is sunlight
 grow in candle extinction jar  chemosynthetic – energy source
• provides an atmosphere is chemicals
with 12-17% oxygen
and 3-5% carbon  Physical Requirement
dioxide 1. Right oxidation-reduction potential of
 eg: Neisseria gonorrhea culture medium
2. Temperature :
VI Nutritional Requirements  Cryophilic
 nutrients • cold-loving (0-20°C)
carbon, hydrogen H2O  Mesophilic
O2 • 20-45°C (most
metabolites organisms)
N2 • most pathogens –
minerals, etc because they grow best
at normal body
o Nutrient source : temperature 37 degree
Celsius
Introduction – Francis Anthony V Marfori MD, BSN 9
Yvonne Agnes Villarta-Marfori MD, BSN

 Thermophilic o function : locomotion


• 45-90°C (hot springs) o the protein (flagellin) is twisted like a
• survive boiling rope
• eg: bacteria with spores o seen in :
 almost all spirals
Gen. Rule :  ½ of bacilli (eg; pseudomonas,
Cold temp  retards growth proteus)
High temp  kills  spirochetes have 2 flagella like
fibrils called axial filaments
3. Light / Radiation o absent in most cocci
4. pH
 prefer a neutral pH  Pili (Fimbriae)
 acidophilic (stomach) – pH 2-5 o hair-like appendage on cell surface
 alkaphiles (intestine) – pH 9 o straighter, shorter, more numerous than
5. Osmotic pressure – concentration of flagella
solutes inside host cell o only in gm(-) bacteria particularly enteric
 Halophilic / Haloduric / cocci
• grow in high o origin : cell cytoplasm
concentration of salt o function :
 Saccharophilic  adherence to specific receptors
• in high conc. of sugar on the human cell surface
6. Moisture infection
 some microorganism can  attachment to other bacteria and
survive complete desiccation viruses
(drying process)  grow and  sex pilus (forms the attachment
reproduce once in a moist between male / female bacteria
environment during conjugation)
• for transfer of genetic
VII Biochemical / Metabolic Activities
material to another
 as bacteria grows  produce many waste bacterial cell by a
products and secretions process know as
conjugation
VIII Pathogenicity o donor is the
“male”
IX Amino Acid Sequencing of Proteins o recipient is the
“female”
X Genetic Compositions
True motility
o bacteria progress continuously in a
given direction

Brownian movement
o organisms all vibrate at the same rate
o due to bombardment of bacteria by
molecules in suspension

2. Bacterial Cell Envelope (cell wall + surface adherents)


Bacterial Cell Ultrastructure:  Surface-adherent materials
1. Append o Glycocalyx (slime layer)
age  a polysaccharide coating
 Flagella  secreted by many bacteria
o whip like filaments from cell wall  allows the bacteria to adhere to
o thread-like protein appendage with a structures like heart valves,
whip like motion
Introduction – Francis Anthony V Marfori MD, BSN 10
Yvonne Agnes Villarta-Marfori MD, BSN

skin, teeth surface forms • rich in mycolic acids


plaque resist decolorization
o Capsule with acid-alcohol
 gelatinous layer
 covers the entire bacterium
 composed of polysaccharide  Plasma membrane
(except anthrax) o lies just inside the peptidoglycan layer
 the sugar component varies and o Functions:
determines the serologic type  waste removal
species  active transport of molecules
into the cell
 role of capsule:
 ATP synthesis
• stimulate Ab formation
 secretion of enzymes/ toxins
• serves as virulence
factor  folds into the cyto during cell
division to form mesosomes
• vaccine preparation
(immunogenic) • mesosome - ensures
the separation of
• used in the specific
chromosomes by
identification of an
forming a cross wall
organism
o In the presence
 Cytoplasm (cell sac)
of homologous
o importance :
Ab, capsule
swellsquellu  center of functional activities
ng rx.  primary site of synthetic
processes
 Organisms with capsule o Structures :
 Nucleoid
• Klebsiella pneumoniae
• single, circular DNA is
• Clostridium perfringens located
• Streptococcus  Plasmid
pneumoniae • smaller molecules of
DNA
 Cell wall
o outermost component common to all
• confer to the bacteria
the ability to produce
bacteria except :
bacteriocin prevents
 mycoplasma (cell membrane)
bacteria overgrowth and
 spirochete determines the type/
o maintains cell shape distribution of bacteria
o phage receptor site flora in the body
o determine gram stain reaction • carry genetic
o peptidoglycan layer: determinant for
 most important component resistance to antibiotics
 responsible for tensile strength / and for synthesis of
shape toxin
• for pili production
 target for antibacterial drugs
 Ribosomes
(pen, cephalosporin)
o site of action of some antibiotics • protein synthesis (site of
antibiotic action)
o cell wall of:
 Granules
 gram (-) bacteria contains:
• storage area for
• endotoxin
nutrients
• β -lactamase (enzyme • stain characteristically
that degrades pen) with certain dyes
 Myco. TB : o Chromatophores
Introduction – Francis Anthony V Marfori MD, BSN 11
Yvonne Agnes Villarta-Marfori MD, BSN

 contain pigments for disintegration


photosynthesis Effects:
o Endospores (spores)  Impair protein
 seen only in bacillus anthracis / synthesis of
clostridia tetani and botulinum infected cell
 Impair capillary
 provide bacteria with means of
permeability
survival in adverse conditions
 Interfere with
 most resistant living structure synaptic activity
known to science owing to its
multiple coverings great 5. extracellular enzymes:
resistance to heat / chem. o coagulase
 destroyed by : steam-heating  in staphylococci
under pressure (autoclave)  enable to clot plasma, forming
 Diseases caused by sticky coat of fibrin around
sporeformers : themselves for protection from
• Anthrax Tetanus phagocytes
• Botulism Gas o kinase (streptokinase)
gangrene  opposite effect with coagulase
 lyse fibrin clot and enables
Attributes of Microorganisms that Enable them to streptococci to invade and
Cause Diseases: spread throughout the body
1. capsule o hyaluronidase
2. flagella  AKA spreading factor
o aid bacteria to invade the aqueous area  enables pathogens to spread
of the body through connective tissue by
o eg: some bacteria from ureter reaches breaking down hyaluronic acid
the kidneys (cement that holds tissue
3. pili (fimbriae) together)
o able to adhere to cells with in mucous  staph, strep and clostrodium
membrane to establish an infection o collagenase
o eg: N. gonorrhea attaches to urethral  breaks down collagen, the
cells and multiply supporting protein found in
4. toxins : endotoxins / exotoxins tendons, cartilage and bones
o hemolysin
Exotoxin Endotoxin  causes damage to the host’s
very toxic Weakly toxic RBC
in the cytoplasm Found in cell wall o leukocidin
does not produce fever Produce fever  enzyme secreted by some staph
Produced by gm (+) / (-) Produced by gm (-) and strep that causes
bacteria bacteria destruction of WBC
A polypeptide excreted by a lipid-polysaccharide
living cell peptide Intermicrobial Relationship:
 when microorganisms live together with other
Destroyed rapidly by heat withstands heating microorganisms a relationship is established
at 60°C which could be:
Can be converted to toxoid cannot be converted to o Symbiosis
by heat/formalin toxoid  a general relationship between
dissimilar organisms living
Highly antigenic  antigenic but does not together in close proximity
stimulate Ab formation stimulate antitoxin  relationship maybe beneficial,
(anti-toxin) formation because Ab is harmless or harmful
against lipopolysaccharide o Mutualism
moiety  a symbiotic relationship where
in both organisms benefit
released only upon cell
Introduction – Francis Anthony V Marfori MD, BSN 12
Yvonne Agnes Villarta-Marfori MD, BSN

o Commensalism o Reduce surface tension of cell


 1 organism derives benefit; membrane
none for the other  Detergent / soap
 eg E.coli in the human intestine
o Synergism  alcohol
 2 organisms accomplish  zephiran
together what neither can
accomplish on its own
 halogens (I2,Cl2) :
• tincture of iodine
 eg: spirochetes + rods • betadine
(fusobacteria)  gum infection  phenols
(trench mouth)
• cresols (wood
preservative)
o Parasitism
 1 organism feeds on the host
and often cause injury to the  hexachlorophene :
host
• soap
 eg: pathogen that cause human • deodorant
disease
o Antagonism
• Phisohex
 presence of 1 organism  hexylresorcinols loosens
prevents the presence of bacte attachment to host cell
another  mouthwash
 eg: gm(+) vs gm(-) : in the respi o Denatures protein :
tract  alcohol
 acetone
Sterilization / Disinfection:  acid/ alkali
 Sterile o Modify protein/ nucleic acid
o state of being free of microorganism of  mercuchrome
any kind o Oxidize free sulfhydril groups
 Septic  iodine
o characterized by the presence of  chlorox
pathogenic microbes in living tissue o Alkylating agents
 Aseptic  substitutes alkyl groups alter
o absence of pathogenic microbes in the chemistry of proteins,
living tissue nucleic acid
 Antiseptic  formalin (production of toxoid)
o substance that opposes sepsis by killing o H2O2
or preventing growth of microorganism
 oxidizes protein
o applied to body surface
o bacteriostatic to harmful vegetative form  creates aerobic environ
prevents growth of aerobes
of organisms
o used at lowest concentration to achieve
2. Physical antimicrobial agents
effect
 Disinfectant  soap dissolves lipids in cell memb
o chemical or physical agents used to kill H2O mechanical removal
microbes on body surface but too toxic  heat
to be applied directly to living tissue o method of choice; most practical,
o applied on inanimate surface efficient, inexpensive and reliable
o used at highest concentration method
o bactericidal o 2 factors to consider
 time
Mechanism of antimicrobial action: (antiseptic and  temperature
disinfectant)  higher the temperature, shorter
1. Chem. agents that damages cell membrane the time
Introduction – Francis Anthony V Marfori MD, BSN 13
Yvonne Agnes Villarta-Marfori MD, BSN

o cause denaturation of protein o kill the pathogens before they mutate


o types and become resistant to it
 dry heat – baking of metals,
glass wares, oils for 2 hours at  Types of antibiotics based on mechanism of
160-165 degree Celsius or for 1 action :
hour at 170-180 degree Celsius 1. Attacks peptidoglycan layer  inhibit cell wall
 moist heat – steaming for 10 synthesis
minutes at 70 degrees Celsius o β -lactam penicillins
or boiling for 10-30 minutes at o cephalosporin
90-100 degrees Celsius 2. Inhibit cyto. memb. function
 pressurized steam – autoclave o polymyxin
(a large metal pressure cooker) o imidazole (antifungal)
– 15psi at 121.5 degree Celsius o nystatin
for 20 minutes 3. Inhibit protein synthesis
 freezing o erythromycin
o good for preserving bacterial culture o chloramphenicol
o the metabolic activities are slowed  o tetracycline
inhibiting their growth o aminoglycosides
 radiation 4. Interfere with nucleic acid synthesis
o UV rays : a. inhibit RNA synthesis – rifampicin
b. inhibit DNA synthesis – quinolones
 for airborne organisms and trimethoprim
 in op room; hosp ward
 ionizing radiation :
• sterilize medicine,
sutures
 however, do not penetrate glass
and building materials, thus
effective only on the air
 filtration
o the disadvantage: small viruses may not
be filtered

Antimicrobial Agents
 compounds/substances produce by
microorganisms or produced by chemical
synthesis that, in low concentration, inhibit
growth of microorganisms
 Bactericidal
o agents with property of killing bacteria
o effect is irreversible
 Bacteriostatic
o agents which inhibit bacte multiplication
o relies on the immune system of the host
to get rid of the organism
o effect is reversible
 ideal antimicrobial agent
o kill or inhibit the growth of pathogens
o cause no damage to the host
o cause no allergic reaction in the host
o stable when stored in solid or liquid form
o remain in specific tissues in the body
long enough to be effective