Disinfection is the elimination of pathogens, except spores, from inanimate objects

Disinfection

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

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

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 10C increase in temperature) Microbe type (e.g., mycobacteria, spores, and certain viruses can be very resistant to disinfectionin general vegetative cells in log phase are easiest to kill) Substrate effects (e.g., high organic content interferes with disinfectionstainless steel bench easier to disinfect than turd) It is easier (and faster) to kill fewer microbes than many microbes

Cleansing is the removal of soil or

Cleansing

organic material from instruments and equipment & may be done, clinically, in four steps:
Rinsing the object under cold water
Applying detergent and scrubbing object Rinsing the object under warm water

Drying the object prior to sterilization or disinfection

Sterilization is the total elimination of all microorganisms including spores

Sterilization

Typically the last things to die are the highly heat- and chemical-resistant bacterial endospores Instruments used for invasive procedures must be sterilized prior to use

Moist heat or steam, radiation, chemicals (e.g., glutaraldehyde), and ethylene oxide (a gas) are employed for sterilization Sterilization by autoclaving, which uses moist heat, is used in most hospital and microbiology laboratory settings

 Sanitization: Lowering of microbial counts to prevent transmission in public setting (e.g., restaurants & public rest rooms)

Other Terms

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)

Exponential Death

10 50C 9

60C

8 0

8060C C

10 Time.

20

30

40

Different Kinds of Bacteria Death

Total cell count

1. Bacteriostatic
Viable cell count

2. Bacteriocidal

3. Bacteriolytic Time

Resistance to Killing

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)

Chemical Antimicrobials

Agent Surfactants Quats (cationic detergent) Organic acids and bases Heavy Metals Halogens Alcohols Phenolics

Mechanisms of Action Membrane Disruption; increased penetration Denature proteins; Disrupts lipids High/low pH Denature protein Oxidizing agent Disrupts cell membrane Denatures proteins; Disrupts lipids Disrupts cell membrane

Comments Soaps; detergents Antiseptic - benzalconium chloride, Cepacol; Disinfectant Mold and Fungi inhibitors; e.g., benzoate of soda Antiseptic & Disinfectant; Silver Nitrate Antiseptic - Iodine (Betadine) Disinfectant - Chlorine (Chlorox) Antiseptic & Disinfectant Ethanol and isopropyl Disinfectant Irritating odor

Aldehydes

Denature proteins

Gluteraldehyde - disinfectant (Cidex); Formaldehyde disinfectant

Used in a closed chamber to sterilize

Ethylene Oxide

Denaturing proteins

Oxidizing agents

Denature proteins

Hydrogen peroxide antiseptic; Hydrogen peroxide disinfectan; Benzoyl peroxide antiseptic

Surfactants

Soap & Detergents

Soaps are sodium or potassium salts of fatty acids, a natural product

Detergents, instead, are artificial surfactants

While soaps are always negatively charged, some detergents are negatively charged while others are positively charged One example of a positively charged detergent are quaternary ammonium compounds (a.k.a., quats)

 Quats are cationic detergents that act by disrupting lipid bilayers

Quats are bactericidal, fungicidal, viricidal (enveloped), and amoebicidal

Quats

Quats are most effective against Grampositive bacteria

Quats do not kill endospores, Mycobacteria spp., nor non-enveloped viruses

Quats are rapidly inactivated by organics including cotton and soap Zephiran Benzalkonium chloride Cepacol Cetylpyridinium chloride

 Ag, Cu, Hg, Ni, Zn, Ag(NO3)2, CuSO4, ZnCl2, HgCl2

Heavy Metals

These metals (and metal ions) react with sulfhydral (SH) groups of proteins, denaturing proteins Silver nitrate is used to treat Ophthalmia neonatorum in newborns as caused by Neisseria gonorrhoeae Oligodynamic action: the ability of very small amounts of heavy metals (especially silver and copper) to exert antimicrobial activity

 Halogens are the seventh (VII) column of the periodic table of elements Two halogens are regularly employed as antimicrobials: Iodine and Chloride

Halogens

Iodine: commonly used as an antiseptic against all microbes, fungi, and viruses
Iodine: It inhibits protein synthesis and oxidizes SH groups of amino acids Chlorine: Used as a disinfectant (10% bleach) Chlorine: Hypochlorous acid (HOCl) is a product, formed in water, that is the active form of the disinfectant Chlorine: Applied in treatment of drinking water, swimming pool, and sewage

Chlorination
1744 discovered in Sweden 1810 identified as an element 1835 first used to control odors 1890s started to be used as a disinfectant 1896 earliest recorded use in experiments on water supplies 1897 used in England to sterilize water mains following typhoid outbreak 1902 first continuous use in water supplies in Belgium 1909 liquid chlorine (compressed gas) became commercially available Subsequent rapid spread in use of chlorine throughout the world WWI: Chlorine gas used as chemical warfare agent

Chlorination
Hypochlorite may either be added directly (i.e., in the What is known as modern chemical warfare began form World War I. The first chemical agent to be used during of bleach) or created within water by bubbling chlorine gas through the water was large amounts of chlorine gas, about one hundred Chlorine gas - preferred for medium to large disinfection sixty tons, which was released from 6,000 pressurized systems cylinders into the wind by the Germans against the Allies. The chlorine floated in a huge clouds for small Sodium Hypochlorite (liquid) - typically used toward the disinfection systems and large swimming pools Allies until it reached the Allied lines causing men to die from the Hypochlorite (powder, tablet) typically of the Calcium effects of the chlorine gas. -Because used largeprivate swimming pools for amounts of gas released the chlorine caused large water purification, do not use scented bleach amounts of yellowish fluid to form in the lungs of For its victim, also causing eye, nose, and throat burning Bromine sometimes by choking. before causing death used as a less-smelly alternative Hypochlorite is less effective in the presence of http://www.geocities.com/CapeCanaveral/Lab/4239/chemweapons/history.html significant organic compounds

 Aqueous ethanol (60-95%) and isopropanol are used as disinfectants Effectively kill bacteria and fungi but not endospores nor nonenveloped viruses

Alcohols

Fast acting, no residue (evaporate away), no staining

But not very penetrating and no residual activity (once gone gone) Exert their action by denaturing proteins and dissolving lipids

In tinctures, they enhance the effectiveness of other antimicrobial chemicals

Flammable; also may damage rubber, plastic, etc.

 Formulae: Fresh juice of Organic Habanero peppers, New Mexico Jalapeno, African Bird peppers and Hatch Chili peppers. Dosage: Five to thirty drops, three times daily. CAUTION ~ EXTREMELY HOT!! Therapeutic Action: Cayenne is the greatest herbal aid to circulation and should be used on a regular basis. The extract is very concentrated and gets into the bloodstream quickly and makes it a perfect first aid remedy for heart attacks, stroke, fainting, shock, dizziness, hemorrhage, internal and external bleeding. Use a few drops to 10 droppers full. It has saved many lives.

Tincture

 Formulae: Fresh Garlic Juice, Goldenseal root, Usnea lichen, Myrrh gum, Pine resin, Echinacea root juice, Tea Tree oil, Kelp, Black Walnut inner hulls, Oak galls and Cayenne pepper in 80% grain alcohol.

Tincture II

Dosage: Generally for external use but can be used in the oral cavity. Soak a cotton swab in the tincture and scrub into the infected area, let air dry. It has a burning sensation. If the wound is tender, just flush it with multiple droppers full of the A tincture is a nonvolatile substance tincture but no need to scrub it in.

(medicine) presented as an been an infected occur Therapeutic Action: There has never alcohol solution, when this formulafun been used. It's excellent for treating e.g., (for has with numerous [sic]) any cut or wound and it is anti-bacterial, anti-viral and antifungal. The tree resins in this formula leave an invisible protective, anti-bacterial coating over the wound. This formula was use on a man in England who had the top of his knee torn off in an automobile accident. In 24 hours it literally glued his knee back together. A nurse from Ireland on the scene said in all the years in the hospital, she had never seen such a severe wound close right up and heal, and with no infections.

Iodine & Iodophores

Phenol, Carbolic Acid, & Phenolics

Phenol (carbolic acid) and derivatives

Affect plasma membrane, inactivates enzymes, and denature proteins

Stable, persistant, and especially effective when dealing with disinfecting materials contaminated with organics but leave residual films, can irritate skin, dont kill endospores, and are corrosive to rubber and plastics Some phenolics are mild enough for use as antiseptics while others are too harsh or otherwise dangerous to be employed on living tissue

Hexachlorophene, Triclosan, Lysol, soap

 HOOH, hydrogen peroxide, is most common

Oxidizing Agents

HOOH is not a terribly effective disinfectant or anticeptic This is because bacteria and body tissues contain enzymes (catalase) that inactivate hydrogen peroxide However, the oxygen released upon inactivation can help oxygenate deep wounds and thus kill strict-anaerobe contaminants, e.g., Clostridium tetani

Ozone and peracetic acid are also oxidizing antimicrobial agents

They exert their effect by oxidizing cell macromolecules (e.g., proteins, DNA, etc.)

Glutaraldehyde is capable of effectiving sterilizationat room temperature, even against endospores, and even in the presence of organics, but achieving sterilization requries many hours of exposure and it is nasty stuff to work with!

Glutaraldehyde

Gaseous Chemosterilizers

Propylene oxide (C3H6O) Chlorine gas (Cl2) Chlorine dioxide (ClO2) Ozone (O3) Ethylene oxide (C2H4O) is used to sterilize heat- or moisture-sensitive items is used for items damaged by heat or moisture is not corrosive, not damaging to delicate instruments, microscopes, disposable plastic instruments and materials permeates porous materials dissipates rapidly from material but is costly, toxic, carcinogenic, explosive, and relatively lengthy process

Agent

Mechanisms of Action
Denatures proteins Denatures proteins Denatures proteins Incineration of contaminants Oxidation & Denatures proteins

Comments
Kills vegetative bacterial cells and viruses Endospores survive 121C at 15 p.s.i. for 30 min kills everything Kills pathogens in food products Used for inoculating loop 170C for 2 hours; Used for glassware & instrument sterilization

Physical Antimicrobials

Moist Heat, boiling Moist Heat, Autoclaving Moist Heat, Pasteurization Dry Heat, Flaming Dry Heat, Hot air oven Filtration

Separation of bacteria Used for heat sensitive liquids from liquid (HEPA: from air)

Cold, Lyophilization (also desiccation)

Cold, Refrigeration Osmotic Pressure, Addition of salt or sugar Radiation, UV Radiation, X-rays Strong vis. Light

Desiccation and low temperature

Decreased chemical reaction rate Plasmolysis of contaminants

Used for food & drug preservation; Does not necessarily kill so used for Long-term storage of bacterial cultures
Bacteriostatic Used in food preservation (less effective against fungi) Limited penetration Used for sterilizing medical supplies Line-drying laundry

DNA damage (thymine dimers) DNA damage

Application of Heat
Heat is frequently used to kill microorganisms Thermal death point (TDP) is the lowest temperature at which all bacteria in a liquid culture will be killed in 10 minutes Thermal death time (TDT) is the length of time required to kill all bacteria in a liquid culture at a given temperature

Decimal reduction time (DRT) is the length of time in which 90% of a bacterial population will be killed at a given temperature (especially useful in canning industry) Dry heat kills by oxidation (slow, uneven penetration)
Moist heat kills by protein coagulation (denaturation) so requires lower temperatures or shorter times, but the moisture must penetrate to pathogens to be effective (grease & oil can block)

Moist Heat
Moist heat kills microbes by denaturing enzymes (coagulation of proteins) Boiling (at 100C, I.e., at sea level) kills many vegetative cells and viruses within 10 minutes Autoclaving: steam applied under pressure (121C for 15 min) is the most effective method of moist heat sterilizationthe steam must directly contact the material to be sterilized Pasteurization: destroys pathogens (Mycobacterium tuberculosis, Salmonella typhi, etc.) without altering the flavor of the fooddoes not sterilize (63C for 30 seconds) Higher temperature short time pasteurization applies higher heat for a much shorter time (72C for 15 seconds) An ultra-high-temperature, very short duration treatment (140C for 3 sec.) is used to sterilize dairy products

Sterilization Times

171o C, 60 minutes, dry heat 160o C, 120 minutes, dry heat 149o C, 150 minutes, dry heat 141o C, 180 minutes, dry heat 121o C, 12 hours, dry heat 121o C, 15 minutes, moist heat (but dont start the clock until entire item is up to tempe.g., large volumes fluid)

Filtration: Air & Fluids

Evaluating Disinfectants

Phenol Coefficient
compares efficacy to that of phenol, with greater efficacy indicated with coefficient >1

Salmonella typhi and Staphylococcus aureus commonly used to determine coefficients Filter-Paper/Disk Diffusion method
placement of disinfectant impregnated filter paper on well-inocated agar Use-Dilution test drying bacteria to surface followed by exposure to disinfectant and subsequent washing and inoculation of sterile broth

Disk Diffusion Method

Hypochlorous acid
Phenol

Nisin Lysol

Escherichia coli

Hands Spread Disease

Link to Next Presentation

Acknowledgements

http://www.sw.vccs.edu/rth/RTH_132/decont.ppt http://www.nt.gov.au/powerwater/html/pdf/tennant_creek_chlorination.ppt http://www.bugcruncher.com/ws/docs/lekcii/D&S_INQUAL_eng.ppt (nice overview of specific antimicrobials) http://www.delmarhealthcare.com/olcs/white/ppts/chapter%2021.ppt (nice overview of microbilogy in nursing) http://www.countyofkings.com/health/ehs/forms/FDHNDLR.ppt http://bioeng.ttu.edu.tw/~chunyi/microbes/Chapter7.ppt http://microbiology.okstate.edu/courses/micro2124/babus/Chpt7_files/chapter7.ppt

Chemical Antimicrobials

* Type of Disinfectant: H = High level; I = Intermediate level; L = Low level