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Medical Microbiology

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43 views6 pages

Medical Microbiology

Uploaded by

tealempo80
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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AL Esraa University College

Subject: Medical Microbiology


Second stage/ 1st semester
Teacher: Assistant Lecturer: Zaineb Aziz AL-Abbassi

Sterilization: is a freeing of an article from all living microorganisms


including bacteria and their spores, viruses, yeasts, molds (pathogenic and
nonpathogenic).
Methods of Sterilization
Physical methods.
Chemical methods
Mechanical methods

Physical methods of sterilization:


A -Heat sterilization
1) Dry heat sterilization
a. Red heat used to sterilize wire loops ,point end of forceps
b. Flaming: used to sterilize mouth of tubes, glass spreaders (which
are flamed in ethanol).
c. Incineration in pathological fuming materials
d. Hot air oven:
(160-180) oc for 2-4 hr., used to sterilize glass wares (pipette)

Conical flask, volumetric flask, Beaker, Graduated cylinder, glass Petri dish

2) Moist heat sterilization


A. Temperature below 100 0 C, pasteurization (63 0 C for 30 min), to
sterilize milk
B. Temperature at 100 o c
Boiling (5-10 min) to sterilize rubber tubes, glass syringes (kills all non-
spore forming bacteria).
Steaming: steam 30 min for 3 days ,used to sterilize gelatin media, sugar
media.

C . Temperature above 100 o c (autoclave) the condition used in this


instrument (121 o c, 15 min, 15 p/inch2 ), used for sterilization of :
surgical tools and clothes culture media and to sterilize
inoculated media

B -Radiation sterilization
Non ionizing type, like ultra-violate rays, infra-red rays
Ionizing type, like Gamma rays, X ray, Beta rays Ultraviolet
Lampe (UV rays)

Gases:
formaldehyde ethylene oxide
Ethylene Oxide: Kills germs by damaging their DNA-RNA
Mechanical methods:
Filtration.: The material is effect by heat (Heat sensitive solutions)
Ex. (serum, protein, sugar, vaccine, antibiotics) are sterilized by filtration

pH
Pressure
• Osmotic pressure
Chemical methods of sterilization
Antiseptic:
It is chemical substance that kill micro-organisms on living tissues, ex.
70% alcohol, heptane, 10% Dettol to sterilize hand.
B- Disinfectant:
IT is a chemical substance used to sterilize non -living objects
ex. Phenol, 5% formalin to sterilize refrigerator
The disinfectant may be described either as:
Bacteriostatic.: any chemical substance which inhibits the growth and
multiplication of bacteria but do not necessarily kill them.
Bacteriocidal: any chemical substance which kills the bacteria and their
spores.
Sepsis: presence of infection (M.O) in living tissue.
Asepsis: Absence of infection (M.O) in living tissue.
Culture Media

Culture Medium is an artificial food prepared in laboratories.


Use of culture media:

l. Isolate the microorganism (bacteria).


2. Identify the microorganism, and study the characteristic of microorganism.
3. Study the antibiotics sensitivity.
4. maintain stock culture.

Basic requirements for bacteria culture media:


1 Energy sources and Nutrition
a. Carbon source: Ex: sugars, carbohydrate, C02
b. Nitrogen source
c. Others (like; Sulfurs, phosphorous, metal Salts, trace elements,
vitamins, essential metabolites).
2 pH (acid, alkaline, natural).
3 02 (aerobic or anaerobic).
4 Time for incubation.
5 Temperature: The optimum temperature of pathogenic bacteria is 37
6 Moisture

Culture Media prepared in suitable containers according to solidity of culture


media
• flasks or tubes with suitable sizes
a. Media dispense in plate Petri-dish for solid media, if colonial
characteristics of an organism are to be examined, the Petri dish is an
excellent container for the medium. The dish should be flat-bottomed, and
either of heat-resistance glass or plastic.

Kinds of media
Culture media are divided into 3 types according their solidity:
1. Solid.
2. liquid
3. Semisolid.
In liquid media the bacteria are free to move, but when grown in
solid media, they multiply at the site of inoculation and form
colonies. The appearance of these colonies is often typical of these
species; this makes possible the isolation of single species of
bacteria from a mixture.

Liquid media are solidified by the addition for example of gelatin or


agar.

Agar is a long-chain carbohydrate, obtained from sea algae it melt at 80


— 100 c o and solidified at 35- 42 co which does not affect the nutrient
properties of the original media (doesn't provided any nutrition to the
bacteria). It acts as solidifying agent.

Gelatin is a protein derived from the collagen of bone, skin

Culture Media Storage: For longer periods (1 week —3 week) store at 4


6 oc in (refrigerator).
Nutrient agar

Blood agar
Chocolate agar

MacConkey Agar
Is selective for Gram negative organisms, and helps to differentiate
lactose fermenting gram negative rods from Non lactose fermenting gram
negative rods. It is primarily used for detection and isolation of members of
family enterobacteriaceae and Pseudomonas spp.

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