Basic Microbiology

Mr. Mohamed Yusuf Abdi

M.Sc. Medical Microbiology and Immunology
 Define Microbiology?
 Micro - too small to be seen with the naked eye.
 Bio – life.
 Logy- means study
So, microbiology is the study of microscopic organisms (or microbes)
 Microbes can be divided into two categories:
A. Cellular microbes (or organisms ,ex. Bacteria, the Archaea, the
Fungi, and the Protists )
B. Acellular microbes (or agents, ex; Viruses, Prions and Viroids)
 Definitions
Taxonomy:
• Science of classifying organisms with goal of showing relationships
among them.
Classification:
• Arrangement of organism into taxonomic groups on the basis of
similarities or relationship.
Nomenclature:
• Naming microorganisms.
 Classification

I. Domain Eubacteria: “True bacteria”.

II. Domain Archaeabacteria: “Ancient bacteria”

III. Domain Eucarya: All eukaryotes: Protista, Fungi, Plantae,

and Animalia.
Prokaryotic Vs Eukaryotic Cells
 What about a virus, is it Prokaryotic or
Eukaryotic and why
• Viruses are not part of the Three Domain Classification, since they are
not considered living organisms.
• Viruses are not considered living organisms by most biologists,
because they lack cells and their own anabolic machinery.
• They are classified separately, using characteristics specific to viruses.
 Bacterial Cell Structures
• Bacteria are procaryotes with free circular DNA, ribosomes, no
mitochondria and a peptidoglycan cell wall.
• Bacterial structure includes:
– internal structure: nuclear material, ribosomes, cell membrane
– cell wall: peptidoglycan (plus periplasmic space and outer membrane
in Gram-negative bacteria)
– external structures: capsule, pili, flagella.
• Spores are metabolically inert in a protective coat.
 Structure of
Bacterial cell
 Cell Wall & Cytoplasmic Membrane
Cell Wall:
• The cell wall is the outermost component common to all bacteria
(except Mycoplasma species) and it is composed of peptidoglycan. It
maintains the characteristic shape of the cell.
Cytoplasmic Membrane:
• Just inside the peptidoglycan layer of the cell wall lies the cytoplasmic
membrane, which is composed of a phospholipid bilayer.
 Unusual and Wall-less Bacteria
• Bacteria belonging to the phylum Chlamydiae appear to lack
peptidoglycan. This has an advantage to the cell in providing
resistance to β-lactam antibiotics (such as penicillin), which attack
peptidoglycan.
• Mycobacteria: contains peptidoglycan but has large amounts of
mycolic acids.
• Mycoplasma: cell wall lacking bacteria.
 Cytoplasm
• Its major component is water (70%–80%).
• Serves as a solvent for the cell pool, a complex mixture of nutrients
including sugars, amino acids, and salts. Cell cytoplasm contents:
ribosomes, nutrient granules, plasmids and nucleoid region.
Ribosomes:
• The ribosomes are the location for protein synthesis.
• Bacterial ribosomes are 70S in size, with 50S and 30S subunits,
whereas eukaryotic ribosomes are 80S in size, with 60S and 40S
subunits.
 Nucleoid
• The genetic material of a bacterial cell is contained in a single, long
molecule of double-stranded deoxyribonucleic acid (DNA).
• The bacterial chromosome is haploid and replicates by simple fission.
• Remember:* no nucleolus , no nuclear membrane and no
mitochondria within bacterial cell.
 Plasmids
• Many bacteria contain small, circular, double-stranded DNA
molecules separate from the chromosome and replicate independent of
the chromosome.
• Functions of plasmids: Plasmids are not essential for host growth and
reproduction they inhibit, but may confer on it certain properties such
as drug resistance, and toxigencity which may constitute a survival
advantage.
 Inclusions
• Bacterial inclusions are generally defined as a distinct structure
located either within the cytoplasm or periplasm of the cell.
• Their role: serve as energy and nutrient reserves
 Spores
• Under conditions of limited supply of nutrition, certain bacteria form
highly resistant, dehydrated forms called spores or endospores. These
endospores are capable of surviving under adverse conditions viz.
heat, drying, freezing, action of toxic chemicals and radiation.
Resistance
• These structures are extraordinary resistant to environmental stresses.
Spores of all medically important species are destroyed by autoclaving
at 120°C for 15 minutes.
• A capsule is polysaccharide layer that completely surrounds some
bacterial cells.
• Flagella: are the organs of locomotion and it is made up of a protein
called flagellin.
• Fimbriae (also termed pili) are thin, hair-like appendages on the surface
of many Gram-negative and it is composed of proteins called pilins.
• Two types (Based on function):
Common pili: The structure for adherence to cell surface.
Sex pili: The structure for transfer of genetic material .
 Classification of bacteria
• Bacteria were initially classified by their
1. Shape
2. Gram stain
3. Oxygen requirements
4. Biochemical and serological characters.
5. Genetics is now being used to establish fundamental relationships.
 Bacterial Cell Morphology

• They have a few basic shapes:

1. Spherical coccus (plural: cocci, meaning

round)

2. Rod-shaped bacillus (plural: bacilli)

3. Spirilla: Spirilla are rigid spiral or helical

forms.
 Gram stain
• The two different cell wall types can be identified in the lab by a
differential stain known as the Gram stain.
• Developed in 1884, it’s been in use ever since.
• After this stain technique is applied the gram positive bacteria will
stain purple, while the gram negative bacteria will stain pink.
Gram Positive Vs Gram Negative cell walls
 Cell Wall of Acid-Fast Bacteria
• Mycobacteria (e.g., Mycobacterium tuberculosis) have an unusual cell
wall, resulting in their inability to be Gram-stained.
• These bacteria are said to be acid-fast because they resist decolorization
with acid–alcohol after being stained with carbolfuchsin.
• This property is related to the high concentration of lipids, called
mycolic acids, in the cell wall of Mycobacteria.
• Note that Nocardia asteroides, is weakly acid-fast.
 Bacterial Growth Conditions
• Microbes are loosely classified into several groups based on their preferred
Oxygen Requirements :
1. Obligate (strict) Aerobic: grow only in the presence of oxygen. These
organisms have enzymes which neutralize the toxic forms of oxygen.
2. Obligate (strict) anaerobes: grow only in the absence of free oxygen. Do
not have enzymes to neutralize toxic forms of oxygen therefore oxygen will
kill these organisms.
3. Facultative anaerobe: grow in the presence or absence of oxygen. Possess
enzymes to neutralize toxic forms of oxygen.
4. Microaerophilic: requires only a small amount of oxygen.
 Bacterial Reproduction
• Bacteria reproduce by binary fission, a process by which one parent
cell divides to form two progeny cells. Because one cell gives rise to
two progeny cells, bacteria are said to undergo exponential growth.
• Generation Time: is the time required for a bacterium to give rise to 2
daughter cells under optimum conditions.
• The doubling (generation) time of bacteria ranges from as
little as 20 minutes for Escherichia coli to as long as 18 hours for
Mycobacterium tuberculosis.
 Bacterial Virulence Factors
• Virulence:
• Measurement for the ability of microorganism to cause disease.
• Many factors determine microbial virulence or its ability to cause
infection and disease. These factors include:
1. Infective dose
2. Adherence factors: The microbial surface structures that mediate
attachment are called adhesins (in bacteria are fimbriae (pili) and surface
polysaccharides)
3. Invasion of host cells and tissues. Invasion refers to ability to
penetrate and grow in tissues.
 Bacterial Virulence Factors

4. Resist host defences: Antiphagocytic factors: toxins like

leukocidins, capsules which protect the bacteria from phagocytes or
Some can survive inside phagocytes after ingestion.

5. Ability to produce Enzymes: ex Hyaluronidases: facilitate the

spread of infection along tissue spaces.

6. Ability to produce Toxins.

 Infective Dose
 Infective dose is referred to the minimum number of microorganism
that is capable of initiating an infection.
 Certain organisms require a relatively small number to initiate
infection (Low infective dose), example: Shigella: vary low (as low as
10 bacteria)
 In contrast, other organisms required in large number to cause
infection (Large infective dose), example: vibrio cholera require about
1,000,000 bacteria to produce infection.
 ID50: Infectious dose for 50% of hosts.
 Number of microbes that will cause a demonstrable infection in 50%
of inoculated test animals.
 LD50: Lethal dose for 50% of hosts
 Number of microbes that will kill 50% of inoculated test animals.
Endotoxin Vs Exotoxin
 Bacterial Pathogenicity
A pathogen:
• A microorganism capable of causing disease.
Pathogenicity:
• The ability of the pathogen to cause disease in an individual.
Infection:
• Multiplication of an infectious agent within the body.
Nosocomial infections:
• Infections that are acquired or develop during a hospital stay.
 Bacterial Pathogenicity
• An iatrogenic infection is an infection that occurs as the result of
medical treatment or procedures.
• For example, Patients who are given immunosuppressive drugs
because they have received a transplant are more susceptible to
infection. Because any infection in such a patient would probably be
the result of the physician-ordered drug therapy, it would be
considered an iatrogenic infection.
 Bacterial Pathogenicity
• Bacteraemia:
• Circulation of bacteria in the blood
• Septicaemia:
• It is the condition where bacteria circulate and multiply in the
blood and produce toxin
• Incubation period:
• Time from infection until occurrence of initial disease symptoms.
• Invasion:
• The process whereby microorganism enter the host cells or tissues
and spread in the body.
• Colonization:
• It refers to the presence of a new organism that is neither a member of the
normal flora nor the cause of symptoms.
 Types of Pathogen

1. Primary pathogens (also called True pathogens):

 Are pathogens that capable of establishing infection and causing disease

in healthy individuals with intact immunological defences.
2. Opportunistic pathogens:
 Are pathogens that cause a disease only in immunocopremised people.
 These pathogens cause disease when defence mechanism of human are
reduced or altered by accident or surgery or therapy or by underlying
metabolic disorder or by an infectious dieses.
 Sources of Infection
• Sources of Infection could be:
A. Human beings: Patient or A Carrier.
B. Animals: zoonotic disease
C. Insects: Vector-borne
D. Soil and water.
E. Food.
 Modes of Infection Transmission
Contact: direct contact OR indirect contact
Inhalation
Ingestion
Cuts and bites, and via arthropods
Congenital (vertical transmission):
• It is transmission of infection from mother to the fetus. Two types:
trasplacental and via the birth canal
Transfusion or transplantation
Foreign bodies
(catheters,
shunts,
prosthetic heart
valves)
• Bacteria cause disease by two major mechanisms:
1. Toxin or enzyme production
2. Invasion and inflammation.
 Septic Shock
• The findings of fever and hypotension are salient features of
septic shock. Additional features include tachycardia, tachypnea, and
leukocytosis (increased white blood cells, especially neutrophils, in
the blood).
• The biologic effects of endotoxin include the following:
1. Fever due to the release by macrophages of IL-1 (endogenous
pyrogen) and IL-6, which act on the hypothalamic temperature-
regulatory center.
2. Hypotension, shock, and impaired perfusion of essential
organs due to nitric oxide–induced vasodilation, TNF-induced
increased capillary permeability, bradykinin-induced vasodilation,
and increased capillary permeability.
3. Disseminated intravascular coagulation (DIC) due to activation of
the coagulation cascade, resulting in thrombosis, a petechial or
purpuric rash, and tissue ischemia, leading to failure of vital organs.
4. Activation of the alternative pathway of the complement cascade,
resulting in inflammation and tissue damage.
 Septic Shock vs Toxic Shock
• Septic shock is different from toxic shock.
• In septic shock, the bacteria are in the bloodstream, whereas in toxic
shock, it is the toxin that is circulating in the blood.
• The clinical importance of this observation is that in septic shock,
blood cultures are usually positive, whereas in toxic shock, they are
usually negative.
 Immunopathogenesis
• In certain diseases, such as rheumatic fever and acute
glomerulonephritis, it is not the organism itself that causes the
symptoms of disease but the immune response to the presence of the
organism.
• For example, in rheumatic fever, antibodies are formed against the M
protein of S. pyogenes, which cross-react with joint, heart, and brain
tissue. Inflammation occurs, resulting in the arthritis, carditis, and
chorea that are the characteristic findings in this disease.
 Bacterial Infections Associated
with Cancer
• The fact that certain viruses can cause cancer is well established, but
the observation that some bacterial infections are associated with
cancers is just emerging. Several documented examples include
(1) the association of Helicobacter pylori infection with gastric
carcinoma and gastric mucosal-associated lymphoid tissue (MALT)
lymphoma.
(2) the association of Campylobacter jejuni infection with MALT
lymphoma of the small intestine.
 Typical stages of an infectious disease
• A typical acute infectious disease has four main stages:
1. The incubation period, which is the time between the acquisition of
the organism (or toxin) and the beginning of symptoms (this time
varies from hours to days to weeks, depending on the organism).
2. The prodrome period, during which nonspecific symptoms such as
fever, malaise, and loss of appetite occur.
3. The specific-disease period, during which the overt characteristic
signs and symptoms of the disease occur.
4. The recovery period, also known as the convalescence period, during
which the illness abates and the patient returns to the healthy state. IgG
and IgA antibodies protect the recovered patient from reinfection by the
same organism.
 Lab diagnosis of bacterial infections
A. Staining:
• Gram stain
• Zn stain
• Spore stain
• Capsule stain
B. Culture
C. Serology
D. Molecular methods
 Normal Flora
• The human microbiome (or human micro biota) is the aggregate of
microorganisms that reside on the surface and in deep layers of skin,
in the saliva and oral mucosa, in the conjunctiva, and in the
gastrointestinal tracts.
• They include bacteria, fungi.
• Some of these organisms perform tasks that are useful for the human
host.
 Origin of Microbial Biota
• Humans acquire the normal flora soon after the birth and then continue
to harbor until death.
• The fetus is in a sterile environment until birth. During delivery
and the first few days of life, the newborn is introduced to the
many and varied microorganisms present in the environment (food,
air).
• As the infant grows, the microbial biota eventually becomes similar
to the microbiota seen in older individuals.
 Categories of Flora
• Microbes typically fall into one of the two categories:
A. Resident flora
• These organisms are life-long members of the body's normal microbial
community.
B. Transient flora
• The transient flora consists of microorganisms that inhabit the body
surface or mucous membranes temporarily for a short interval.
 Beneficial Effects Normal Microbiota
1. Prevent colonization of pathogen: by competing for attachment
sites or for essential nutrients
2. Waste produced antagonize other bacteria: by producing a
variety of waste substances such as: Fatty acids and peroxides,
Lactic acid: Lactobacilli present as normal flora in vagina of adult
females maintain the acidic pH by producing lactic acid, thereby
prevent the growth of pathogenic bacteria
 Beneficial Effects Normal Microbiota
3. Synthesize vitamin: human enteric bacteria secrete
several vitamins such as vitamin K and B complex
4. Stimulus for the development of the immune system.
5. Assist in digestion
6. Play a role in toxin degradation
7. Act as anti-carcinogenesis
 Harmful Effects of Normal Microbiota
A. May be agents of disease: Members of the normal flora may cause
various endogenous disease When the host immunity is lowered,If
they enter a wrong site or tissue (e.g. blood, sterile body cavities)-then
even the resident flora can produce disease. For example, E. coli
which is a resident flora of intestine may cause urinary tract infection
if enters into urinary tract.
B. Bacterial synergism: Bacterial vitamins and growth factors produced
by members of the normal flora may promote the growth of the
potential pathogens
C. Competition for host nutrients: Bacteria in GIT absorb some of the
host's nutrients for their survival.
Disturbed Normal Flora Promote Infection
• When the composition of normal flora is disturbed, it facilitates
pathogenic organisms to enter and cause disease.
• Several mechanisms by which the normal flora is disturbed are as
follows:
A. Injudicious use of broad spectrum antimicrobial agent: It may
completely suppress the normal flora thus permitting the pathogen
to take the upper hand and cause infection. For example,
Clostridioides difficile, causing pseudomembranous colitis.
B. Immune suppression or imunodefisioncy
C. Physical destruction of the normal flora: by irradiations,
chemicals, burns, etc
C. Minor trauma in mouth (e.G. By dental procedure, chewing or
vigorous brushing) can promote passage of small numbers of
bacteria (e.G. Viridians streptococci) transiently into bloodstream,
which can cause bacterial endocarditis.
Viruses
Main properties of viruses:
1. Viruses do not have a cellular organization
2. They contain only one type of nucleic acid, either DNA or RNA.
3. They are obligate intracellular parasites.
4. They lack the enzymes necessary for protein and nucleic acid
synthesis and are dependent for replication on the synthetic
machinery of host cells
5. They multiply by a complex process and not by binary fission
6. They are unaffected by antibacterial antibiotics.
 Structure and Chemical Composition of
The Viruses
• Viruses vary in their structure.
• A virus particle consists of:
A. Viral capsid
B. Viral Envelope
C. Viral Nucleic Acids
 Baltimore classification of viruses based on
replication
1. Double stranded DNA viruses: example: hepadnaviruses.
2. Single stranded DNA viruses: example, parvovirus
3. Double stranded RNA viruses: example Reoviruses
4. Single stranded rna viruses: are classified into two categories
I. Positive strand (plus strand, positive sense): the viral RNA itself act
as the mRNA. example Picorna-virues
II. The negative strand (minus sense) RNA viruses: example rhabdo-,
orthomyxo virus
Baltimore classification of viruses based on
replication
5. Retroviridae exhibit a unique replicative strategy. Their single
stranded RNA genome is converted into an RNA: DNA hybrid by
the viral reverse transcriptase enzyme.
 Virus Replication
• Viral infection of host cells Viral replication in host cells involves the
following steps:
1. Attachment (or ‘adsorption’) of the virus to cell surface molecules
(receptors).Many viruses have highly specific receptors, which
limits the range of cell types that can be infected.
2. Penetration of the virus into the host cell (often by endocytosis).
3. Uncoating follows, which involves the enzymatic removal of viral
protein coat and liberation of nucleic acid and attached core
proteins.
 Virus Replication
4. Production of virus-specific mRNA, in order to direct the host cell
ribosome to produce viral proteins (core, capsid).
5. Morphogenesis and maturation occur with assembly of
components (nucleic acid and proteins) to form viral particles.
6. Release of the virus is by bursting of infected cells (lysis) or by
budding through the plasma membrane.
• Note: Replication of DNA viruses occurs in host cell nucleus, where
in RNA viruses occurs in the cytoplasm.
 Virus Cultivation
• Viruses are obligate intracellular parasites and can therefore replicate
only in living cells.
 Major types of viral infection
Three major types of viral infection occur:
1. Abortive, with no replication, no visible host cell effects, no disease.
2. Cytolytic, with cell death and virus dissemination, then disease, then
death or recovery of the host.
3. Persistent, which is of three further sub-types:
A. Latent, with no apparent effect on the cell, but may re-activate, e.g.
herpes.
B. Productive, which may give chronic carriage or disease e.g.
hepatitis B.
C. Transforming producing tumours e.g. EBV lymphomas.
 Host cell morphological effects due to
viral infection could be
1. No morphological change, in abortive or latent infection.
2. Multi-nucleated cells by fusion
3. Cytopathic effects (CPE) including rounding or darkening of the
cell, or inclusion bodies in the nucleus, or in the cytoplasm, or actual
cell death.
4. Haemagglutination, by viral surface haemagglutinins adhering to
red blood cells.
5. Transformation to malignant cells e.g. by papillomaviruses, EBV,
Hepatitis B or C viruses.
 Laboratory diagnosis of viral
Disease

Direct detection indirect detection

Serological diagnosis
of virus of virus

Detection of
Detection of Egg Animal
Microscopy viral genetic Tissue culture
viral proteins inoculation inoculation
material
 Prions
• Prions (proteinaceous infectious particles) are infectious particles
composed solely of protein with no detectable nucleic acid.
• Unlike viruses, they are highly resistant to inactivation by heat,
formaldehyde, and ultraviolet light that inactivate viruses.
• Prion diseases: Prion diseases, called “transmissible spongiform
encephalopathies,’’ include scrapie in sheep, mad cow disease in
cattle, and kuru and Creutzfeldt–Jakob disease in humans.
• Prions do not appear to be viruses.
 Fungal cell structure
• Fungi are eukaryotic organisms have true nuclei with definite
nuclear membrane, nucleolus, cytoplasmic organelles (including
mitochondria, golgi apparatus, endoplasmic reticulum,
lysosomes).
• Fungal cell wall consist primarily of chitin, thus fungi are
insensitive to antibiotics such as penicillin, that inhibit
peptidoglycan synthesis.
• Fungal cell membrane consist of ergosterol, in contrast to the
human cell membrane which contain cholesterol.
 Fungal Morphology
1) Filamentous fungi (molds):
• Molds: are multi-celled fungi, composed of filament structure,
termed hyphae. Intertwined mass of hyphae is called a
mycelium
• The hyphae may be branched or unbranched.
• Some hyphae contain walls (septate hyphae) where others do not
form walls (non septate hyphae or coenocytic).
 Molds

Septate hyphae Non septate hyphae

 Morphology
2) Yeast: are unicellular round or oval bodies and some are capsulated.
3) Yeast-like fungi : this form of undetached budding yeast-cells
which present the appearance of broad septate hyphae.
 Morphology
4) Dimorphic fungi :
The dimorphic fungi have two forms and this depend on temperature:
1. Yeast (parasitic or pathogenic form). This is the form usually seen in
tissue sections, in some exudates, or if cultured in an incubator at 37ºc.
2. Filamentous (saprophytic form). The form observed in nature or when
cultured at 25ºc.

So in the body, they are in yeast form in the pathologic state.
 Fungal Reproduction

Filamentous fungi (molds) reproduce by:

Asexually by fragmentation of hyphae

Asexual and sexual reproduction by spores.

Yeasts reproduce by:

Asexually by budding or fission

 Source of infection & Mode of transmission

Source of infection:
Endogenous: Normal flora
Exogenous: Source of human infection may be from soil (geophilic),
from animals (zoophilic), or from human (anthrophilic).
Mode of transmission
Respiratory tract (air borne infection)
GIT (food & water borne infection).
Skin = contact or inoculation
 Fungal Virulence factors
• There is evidence that some fungi do have virulence factors:
Ability to adhere to host cells by way of cell wall glycoproteins
Production capsules allowing them to resist phagocytosis
Production of a cytokine that suppress the production of complement.
Ability to damage host by secreting enzymes such as keratinase, elastase,
collagenase.
Ability to secrete mycotoxins (ex. Aflatoxin)
Ability to block the cell-mediated immune defences of the host.
Chronic fungal infections can also provoke an allergic response in the host.
 Types of Mycotic Diseases
1. Allergies or Hypersensitivity to fungal spores, particularly aspergillus
spore which cause asthma.
2. Mycotoxicosis. The best- known mycotoxicosis occurs after eating
Amanita mushrooms.
3. Infection, and has four types:
A. Superficial mycosis.
B. Cutaneous and Subcutaneous mycosis
C. Systemic mycosis
D. Opportunistic mycosis
Sterilization and
disinfectant
 Terminology

• Sterilization: a process that destroys all viable microbes, including viruses and
endospores.

• Disinfection: a process to destroy pathogens, (except endospores).

• Sanitization: any cleansing technique that mechanically removes microbes.

• Antiseptics: Chemicals that can be safely applied on skin or mucous membrane to
prevent infection.

• Decontamination: Process that removes pathogenic microorganisms from an

object to make it safe to handle.

• Disinfectant: Products used to kill microorganisms on objects or surfaces.

Disinfectants are not necessarily sporicidal, but may be sporostatic, inhibiting
germination or outgrowth.
 Methods of Sterilization
1. Physical methods
A. Heat:
• Dry
• Moist
B. Radiation:
• U.V. light
• Ionizing radiation
C. Filtration
2. Chemical methods:
A. Phenol Derivatives : Phenol, Cresol, resorcinol, chloroxylenol
B. Oxidizing agents :Pot.Permanganate
C. Halogens : Iodine, chlorine
D. Biguanide : Chlorhexidine
E. Alcohols : Ethanol, Isopropanol.
F. Aldehydes : Formaldehyde, Glutaraldehyde
G. Acids : Boric acid, acetic acid
H. Metallic salts ; Silver Nitrate, Zince Sulfate, Zinc Oxide.
I. Dyes : Gentian violet, proflamine, Acriflamine.
 Heat
• Most common and one of the most effective methods of sterilization.
• Factors influencing sterilization by heat are:-
I. Nature of heat :
a. Dry
b. Moist
 A. Dry Heat
1. Red Heat: It is used to sterilize metallic objects by holding them
in flame till they are red hot. Example: inoculating wires and
forceps (metallic objects).
2. Flaming: is passing an object over the flame without allowing
it to become red hot. (used to sterilize glass slides).
• 3. Hot air oven :
• It is used to sterilize items, which do not get damaged by high temp.
such as glass, metallic objects, wooden stick and Powders.

Time(Min) Temperature
180 140oC
150 150oC
60 160oC
45 170oC
18 180oC
7.5 190oC
 Moist heat
1. Pasteurization :
• The temperature employed is either 630C for 30mins (Holder method)
or 720C for 15-20 seconds (Flash method) followed by cooling
quickly to 130C.
• Method is used for heat sensitive liquid and pharmaceutical products.
2. Autoclave
3.:steam is the effective of sterilization, because of its:
1. High penetrating capacity.
2. It gives of large amount of heat to surface with which it comes in contact.
• Autoclaves essentially consist of following:
I. A cylindrical or rectangular chamber.
II. Water heating system
III. Steam outlet and inlet valves
IV. Single or double doors with locking mechanism.
V. Thermometer
VI. Pressure gauges
• To achieve sterility, a holding time of at least 15 minutes at
121 °C at 15 psi or or 10 minutes at 126 °C at 20 psi.
 3. Irradiation
• Radiation used for sterilization is of two types:
1.Ionizing radiation, e.g., X-rays, gamma rays, and high speed electrons .
2.Non-ionizing radiation, e.g. ultraviolet light, and infrared light.
• These forms of radiation can be used to kill or inactivate
microorganisms.
Thank You