STERILISATION

Sterilisation is a process by means of which an

article, surface or medium is made free from all
living micro – organisms including spores.
Antiseptics – substances which either kill micro –
organisms or inhibit their growth.

Disinfection – compounds that kill micro – organisms

(except bacterial endospores). Since these may be
harmful to human tissues , disinfectants are usually
reserved for inanimated objects.
Sterliant – chemical agents that are destroy all micro
organisms , including spores.

Germicide – chemical or physical agent that destroy

most organisms , but not endospores , used on skin or
inanimate objects.

Sporocide – generally a chemical agent that destroy both

bacterial and fungal spores.
Fungicide – chemical substances that destroy virus.

Virucide – chemical agents that destroy viruses.

Bacteriostat – chemical agents that inhabit bacterial

growth.

Sanitizer – an agent , usually a detergent , that reduces

the number of bacteria to a single level.
 Method Concentration(level of activity)
A. Physical sterilants

1. Steam under - 121’C for 15-20min or pressure

134’C for 3min.
2. Dry heat – 171’C for 1hrs; 160’C for
2hrs; 121’C for 16hrs.
3. Filtration – 0.22 to 0.45Nm pore size;
HEPA filters
4. UV radiation – exposure at 25.4nm
wavelength for variable time
5. Iodinizing - exposure to
radiation microwave or gamma
radiation for variable time.
B. Gas vapors sterilants
1. Ethylene oxide - 450-1200 mg/L at 29` to
65`C for 2 to 5 hrs.
2. Formalchehyde - 2% to 5% at 60` to 80`C
vapour
3. Hydrogen - 30% at 55% to 60`C
peroxide vapour
4. Plasma gas - highly ionized H2O2 gas
5. Chlorine dioxide - variable
gas
C. Chemical sterilants
1. Glutaealdehyde- 2%
2. Peracetic - 0.2%
STERILISATION BY HEAT
1. By dry heat.
2. By moist heat.

1. dry heat – it kills the microorganisms by

destructive oxidation of intracellular
components.
2. Moist heat – moist heat kills the micro – organisms by
coagulating and denaturing their enzymes and
structural proteins.
Procedure of sterilization by dry heat
1. Red heat - metallic objects such as innoculating
wires , tips of forceps and needles are held in the
flame of a Bunsen burner for instant sterilisation.
2. Flaming – certain articles like glass slides , needles ,
cotton – wool plugs are passed over flame. These are
not allowed to get red heat. This method is of
uncertain efficacy.
3. Incineration – this method is employed for destruction
of infective materials. The soiled dressing , bedding ,
pathological materials like sputum and stool and
carcasses are reduced to ashes by burning.
4. Hot air oven – it is the most widely used sterilizer of
dry heat particularly for glassware. Sterilisation of
articles by exposure to hot air is the accepted method
which cannot be reliably penetrated by steam and can
with stand temperature (160`-180`C).
 Procedure of sterilisation by moist heat
(a) At a temperature below 100`C :-

1. Vaccine bath
2. Pasteurization of milk
3. Fractional sterilisation
(b) At a temperature of 100`C :-
1. Boiling at 100`C.
2. Steam at atmospheric pressure at 100`C for
90minutes.
3. Steaming at 100`C for 3 successive days.
(c) At the temperature of above 100`C
Autoclave :- Autoclaving is the process of sterilisation by
statured steam under high pressure above 100`C.
essentially it is a modified pressure cooker or boiler
which may be horizontal or vertical. It is a double –
walled or jacketed chamber made of stainless steel or
gunmetal with a supporting frame. The steam circulates
within the jacket and is supplied under high pressure to
the closed inner chamber. Where goods are kept for
sterilisation.
Types of autoclaves
1. Simple iron – jacked
2. Low – pressure low temperature type.
3. High – pressure high vaccum type where as much as
98% of the air present is rapidly expelled by an
electric pump and sterilisation is done without delay.
 Sterilization by chemical methods :
 Characteristic of ideal antimicrobial chemical agent

1. Antimicrobial activity

2. Solubility

3. Stability

4. Nontoxic to humans and other animals.

5. Homogeneous

6. Non combination with extraneous organic material

7. Toxicity to microorganisms at room or body temperature.

8. Capacity to penetrating

9. Non corroding and non-staining

10. Deodorizing

11. Detergent capacity

12. Availability

13. Example : Phenol, Alcohol, Halogen and Chlorine

 Biological agent:
 Chemotherapy -> Treatment of the disease with chemical
substance is known as chemotherapy

-> And chemical substance used is known as

chemotherapeutic agent.

-> some of these chemical are synthesize in

laboratory .They are known as synthetic therapy.
-> While other are produce by microorganism are
known as natural chemotherapeutic agent.
-> Few are prepared by synthetic.
-> most of prepared by microbial synthesis.
-> Substance must have selective toxicity for
parasites.
-> Substance should damage the parasite without
causing any damage to host
-> Phenol and heavy metals are not used.
STREPTOCOCCUS
The genus streococcus includes a large number of species
of gram – positive , catalase negative cocci arranged in
chains of varying length and also in pairs. They form
chains during growth due to successive cell divisions
occurring in one plane. Some of them form part of the
normal flora of man and animals , some others are
important human pathogens causing pyogenic infections.
1. Streptococcus pyrogens.
2. Streptococcus viridans.
3. Streptococcus faecalis.
Morphology
The individual cocci are spherical o ovoid. Some members
show a diplococcal appearance , and rod – like forms are
occasionally seen. The lengths of chain vary widely with
cultural conditions. Larger chains are formed in liquid than in
solid media. Streptococci are nonmotile and nonspring. Most
strains belonging to group A and C possess capsules.
Culture
They are aerobes , facultative anaerobes and grow best
at a temperature of 37`C , group D streptococci grow
well at and between 10`C and 45`C. growth is poor on
solid media or broth. They grow well in media
containing blood and sugars and 10% carbon dioxide.
Classification
streptococcus

Aerobes and facultative anaerobes obligate anaerobes

peptostreptococci

1.Alpha-haemolytic classified 2. Most beta-haemolytic and 3.Gamma-haemolytic

Into species by physiological some alpha-haemolytic $ classified into
And bio-chemical properties nonhaemolytic species by physio
seriological ( by cell wall) -logical $ bio-
chemical properties
20 lancefield groups(ABCDEFGHKLMNOPQRSTUV)

Group A- streptococcus pyogens

serological typing (M protein)

Griffith types(1,2,3,etc up to 80)

1. Alpha -haemolytic streptococci exhibits
incomplete haemolysis which imparts a greenish
discolouration around the colony.The species
includes viridans group.

2. Beta-haemolytic streptococci exhibit a wide

zone haemolysis.They account for the majority of
streptococcal diseases,although all beta-haemolytic
streptococci are not pathogenic.

3. Nonhaemolytic streptococci include S.faecalis.

 Biochemical reaction
 Most sugars are fermented by streptococci with
production of acid but no gas.They are catalase
negative.Major fermentive product is lactic acid
which accumulates in culture media.
 Clinical syndromes
1. Acute infections of respiratory tract
(a) Sore throat is the commonent of streptococcal
diseases.
(b) Scarlet fever : Scarlet fever occurs as a complication
of streptococcal infection when the infecting strain
produces erythrogenic toxin and the patient has got no
antitoxic immunity.
2. Skin infections
(a) Impetigo : Impetigo is a superficial discrete crushed
spot specially in children,usually less than one inch in
diameter.It last for 1-2 weeks and heals spontaneously
without leaving any scar.
 (b) Erysipelas :Erysipelas is an acute spreading
lesion.Infected area of skin shows massive brawny
oedema with erythema.
(c) Cellulitis : Unlike erysipelas,cellulitis involve
deeper subcutaneous tissues.

3. Acute rheumatic fever-swelling of joints and

pancarditis

4. Acute post-streptococcal glomerulonephritis-

Haematuria,albuminuria,oedema
Laboratory diagnosis
A. Acute suppurative infections : specimen
is collected from the site of lesion such as
swab,pus or blood depending on the nature
of infection.
1. Smears :- Microscopy of smears of pus
showing gram-positive spherical or oval
cocci in chains or pairs in association with
pus cells suggest the presence of
streptococci.
2. Culture
3. Antigen detection