Baedah MadjidBAGIAN MIKROBIOLOGI FK-

UNHAS2007

● Sterilization: total inactivasi segala bentuk kehidupan → kemampuan bereproduksi● Bactericide: agen yang membunuh bakteri ● Bacteristatic: substance yang mencegah pertumbuhan bakteri,● Disinfectant (germicide): agent membunuh mikroorganisme yang mempu menimbulkan infeksi.● Antiseptic: opposes sepsis or putrefaction dengan membunuh bakteri atau menghambat pertumbuhannya. (hidup dalam jaringan)● Decontamination: get rid of contamination (beberapa mikroorganisme.

Agen fisik1. Panas - panas lembab - panas kering2. Pembekuan 3. Radiasi - radiasi ultraviolet - radiasi ionisasi4. Ultrasonic & Sonic getaran5. Filtrasi

Agen kimiaMechanisms of action: ● merusak sell ● membran ● Denaturasi Protein● Modify Functional Groups of Proteins and Nucleic Acids.

Faktor yang mengakibatkan potensi Disinfectant

● Concentration (Cnt = K)● Time of Exposure● pH● Temperature● Nature of the Organisms● Presence of Extraneous materials → organic matter: serum, blood, pus, etc.

6

5

4

3

g10 jmlkteri 2ng bertahandup setelah 1manasan

0 D121

-11 2 3 4 5 6 7

Lama pemanasan pada 121oC

Campuran Name Effects Practical Applic Surface disinfectantsCATIONIC AGENTSQuaternary

ANIONIC AGENTS

NON-IONIC

ZephiranCeepryn chlorideDuponal LSTriton W-30Tween-80

Gram-positive (mostly)Gram-positive (Only)

Antiseptic

Phenolic Cresol

Diphenyl compound

LysolCreolinHexachlorophene

Bactericidal

Gram-positive

DisinfectantGermicidal soapDisinfectantGermicidal soap

Alcohols Ethanol 70% Bactericidal + spores

Disinfectant

Compounds Name Effects Practical Applic.Acids Hydrochloric acid

Benzoic acidLactic, citric, acetic, propionic acids

Antibacterial

DisinfectantFood preservativeFood preservative

Alkalies Hydroxide (metallic cation)

Direct toxic Disinfectant

Alcohol Ethanol 70% Bactericidal + spores

Disinfectant

Acetone Acetone Disinfectant

Free H+ & OH- ions → undissociated molecule or altering pH of organisms environment

Compounds Name Effects Practical Applic . Heavy metals

Mercurials

Silver compound

Mercuric chlorideMetaphen, Merthiolate, Mercurochrome

Phenylmercury saltsSilver nitrate 1%

Very toxicLess toxicGram-pos & neg, fungi, yeast, algaeBactericidal Agents

Limited useAntiseptics

Control contami-nation of Pseudo-monas & others Prophylaxis of ophthalmia neona-torum

Oxidizing HALOGENS

IodineChlorine

HYDROGEN PER-OXIDE

Betadine, Jodium ticture (Sunklin)H2O2 3%

GermcideGermcide

Damage DNA

Skin DisinfectantDisinfectant

Food preservativeWound cleaning

Compounds Name Effects Practical Applic .

DyesTriphenylmethae

dyesAcridine dyes

Malachite greenCristal violetProflavine

Gram-positiveGram-positiveSynthesis of Nucleic acid

Staining, mediumStainingWound antiseptics

Alkylating agents

Formadehyde

Glutaraldehyde

Ethylene oxide

Formalin (sol. 3%)Paraformaldehyde (solid 91-99%)Gas

Gas

All organisms(Irreversible)

10 x formal-dehyde All bacteria

DisinfetantLatex gloves

For rooms, fabrics, instr.Disinfectant forEndoscopic instr.

A. Dry heat a. Hot air → oven: metal/glass, powders, oils b. Flaming : loops/öse, glass object c. Incineration → incinerator: medical trash B. Moist Heat ● Boiling water : 100oC ● Steaming: 100oC ● Autoclaving: 125oC, 15 atmosphere ● Pasteurization: 72C for 15 seconds → milk ● Tyndalization

Moist Heat Dry Heat

Temperature Time (min ) Pressure Time (min)

121oC 15 15 -126oC 10 20 -134oC 3 30 -140oC - 180

150oC - 150

160oC - 120

170oC - 60

protein denaturation & coagulation

► Freezing : preserving► Repeated freezing & thawing → bacterial destruction.► In freezing: formation of ice crystals outside the cells → withdraw water from the cell interior → intracellular electrolyte concentration ↑ & protein denaturation → damage of cell membrane.

A. Ultraviolet radiationB. Ionizing radiation

● Sunlight : spontaneous sterilization occur under natural condition. ● Effects of Radiation: inactivation of molecule by rays energy absorbed by molecule. ● UV: bactericide ● Energy content of Ionizing radiations >>> than UV radiation

Lethal & mutagenic agent●Lethal dose: for non-spore-forming bacteria 1800 – 6500 μW/cm2. Bacterial spores required 10 X more●Bactericidal effect: wavelength of 240-280 nm wavelength of 260 nm lethal but can not penetrate the glass or water.●Practical use: to control air-borne infection → for operating rooms or hospital wards, biological safety cabinet (laminar flow)

► Lethal effects: most pathogenic non-spore-forming bacteria► Products by : a. radioactive decay: α, ß, γ rays b. x-rays, c. particle bombardment d. Nuclear reactor► Practical applications: - Sterilizing dose : 2.5 Mrad → most resistance microorgn - γ radiation of cobalt 60 : pharmacy and medicine, catgut, nylon sutures, disposable medical and medical supplies.

● Sound vibration (20 to 1000 kc) → disrupting cells.● Ultrasonic waves more effective as the frequency is increased. ● Effects: Disintegrating cells & H2O2 formation Depolymerization of macromolecules● The most susceptible: gram negative rods● The most resistance Staphylococcus : require long period exposure.● No practical value for sterilization & disinfection : numerous survivors.

►Sterilization: heat-labile materials (lab)►Mechanisms: ● Electrostatic & absorption phenomena ● Physical construction of filters►Types: ● Old types : Berkefeld, Chamberland, Seitz ● Membrane filters : porous disks of cellulose

● Pore size: 14-0.023 µm.● Most widely used: 0.023 µm. ● Practical used: serum, plasma, etc.●Two-dimensional screen.● Mechanisms: a. Particles > pore : retained on filter surface b. Particle < pore: retained by van der Waals forces.