Sterilization

1. Sterilization NorazliGhadin

2. Objectives • Understand and utilize correct sterilization and disinfection techniques • Distinguish between sterilization and disinfection • List the characteristics of an ideal antiseptic • Describe sterilizing agents and rank their effectiveness • Discuss the time/temperature relationship in destroying microorganisms

3. Disinfection is the elimination of pathogens, except spores, from inanimate objects • Disinfectants are chemical solutions used to clean inanimate objects (physical processes, e.g., UV radiation, may also be employed to effect disinfection) • Germicides are chemicals that can be applied to both animate (living) and inanimate objects for the purpose of eliminating pathogens • Antiseptics are formulated for application to living tissue Destruction/Removal of Harmful Microorganisms Disinfection

4. The Ideal Disinfectant • Resistant to inactviation • Broadly active (killing pathogens) • Not poisonous (or otherwise harmful) • Penetrating (to pathogens) • Not damaging to non-living materials • Stable • Easy to work with • Otherwise not unpleasant

5. Disinfectant Performance… • Is dependent on Disinfectant concentrations • Is dependent on length (time) of administration • Is dependent on temperature during administration (usual chemical reaction 2x increase in rate with each 10°C increase in temperature) • Microbe type (e.g., mycobacteria, spores, and certain viruses can be very resistant to disinfection—in general vegetative cells in log phase are easiest to kill) • Substrate effects (e.g., high organic content interferes with disinfection—stainless steel bench easier to disinfect than turd) • It is easier (and faster) to kill fewer microbes than many microbes

6. Sanitization: Lowering of microbial counts to prevent transmission in public setting (e.g., restaurants & public rest rooms) • Degerming: Mechanical removal of microbes, e.g., from hands with washing • Sepsis: Bacterial contamination • Antisepsis: Reduction of or Inhibition of microbes found on living tissue • Germincides, Fungicides, Virucides • Physical versus Chemical disinfectants • Static (stasis) versus Cidal (e.g., bacteriostatic versus bacteriocidal) Other Terms

7. Gram-negative bacteria (with their outer membrane) are generally more resistant than gram-positive bacteria to disinfectants and antiseptics • Stationary-phase (I.e., non-growing) bacteria generally are more resistant than log-phase (I.e., growing) bacteria • Mycobacteria, endospores, and protozoan cysts and oocysts are very resistant to disinfectants and antiseptics • Nonenveloped viruses are generally more resistant than enveloped viruses to disinfectants and antiseptics • Organic matter (such as vomit and feces) frequently affects the actions of chemical control agent • Disinfectant activity is inhibited by cold temperatures • Longer application times are preferable to shorter • Higher concentrations, though, are not always preferable to lower concentration (e.g., alcohols) Resistance to Killing

8. Chemical Antimicrobials

9. Physical Antimicrobials

10. Moist Heat • Moist heat kills microbes by denaturing enzymes (coagulation of proteins) • Boiling (at 100°C, I.e., at sea level) kills many vegetative cells and viruses within 10 minutes • Autoclaving: steam applied under pressure (121°C for 15 min) is the most effective method of moist heat sterilization—the steam must directly contact the material to be sterilized • Pasteurization: destroys pathogens (Mycobacterium tuberculosis, Salmonella typhi, etc.) without altering the flavor of the food—does not sterilize (63°C for 30 seconds)

11. Sterilization Times • 121oC, 15 minutes, moist heat (but don’t start the clock until entire item is up to temp—e.g., large volumes fluid)

12. Filtration: Air & Fluids

13. Filtration Membrane filters • These are porous membrane about 0.1mm thick, made of cellulose acetate, cellulose nitrate, polycarbonate, and polyvinylidene fluoride, or some other synthetic material. • The membranes are supported on a frame and held in special holders. • Fluids are made to transverse membranes by positive or negative pressure or by centrifugation.

15. USES • Remove toxin from bacterial cells • Sterilize drugs (antibiotic) • sterilize protein, glucose or other sugars • Sterilize media with serum/heat sensitive substance • Studyvirus size •  separate virus from cells

16. RADIATION • Energy transmision through a space ruangan • Basicly all radiation can cause a death/mutation on microorganisme

17. EFFECT OF RADIATION • Changes • Cells membrane • inhibit DNA replication

18. Types of Radition • Ultraviolet (UV) • Ionizing radiation

19. UV • Naturally from the sun. • MoA; • Induction of dimer thymine on DNA chain (mutation):  inhibit DNA replication cell death. • not all organisme can be kill. • Wavelength used (260 nm)

20. UV: MERCURY LAMP • 240 – 280 nm • To decrease bacterial volume in atmosphere, operation theatre, nursery, restaurant & food factory. • weakness : Low penetration power. • Stop by glass, paper, dust or stain • Can only penetrate a few milimeter of solution.

21. UV • Adjacent bases bond with each other, instead of across the “ladder.” • makes a bulge, and the • distorted DNA molecule. • Cant replicate MEDA 1012:MEDICAL MIKROBIOLOGI

22. IONIZING RADIATION • X-RAY and gamma radiation (). • More effective than UV. • MoA: destroying by mutation or oxidation

23. Cont… • Efective against SPORA. • Source : isotope Cobalt 60. Dos 2.5 mrad. • sterilizing disposable: ·  Syringe and needles ·  Chateter ·  heat sensitive items • EXPENSIVE