Asepsis, Sterilization and Disinfection

Pradeep B Patil

Sushruta 1500BCE

Hippocrates (460-377 BC)

Separated medicine from philosophy Irrigated wounds with wine or boiled water Foreshadowing of Asepsis

Joseph Lister
chemical antiseptics to control surgery related infections
disinfect surgical equipment and supplies

Microorganisms are simple forms of life commonly made up of a single cell only seen with a microscope.

Pasteur (1862)
Discovered bacterium Developed Pasteurization process

Mikulicz-Radecki (1887)---face masks Furbringer (1889)-----------hands-scrubbing Halsted (1890)---------------surgical-gloves

Basic terms
Sterilization Disinfection

Antisepsis
Bacteriostasis Asepsis

Microbial Control
Degerming: Removing microbes from a limited area Sanitization: Lowering microbial counts on eating utensils Biocide/germicide: Kills microbes Bacteriostasis: Inhibiting, not killing, microbes

Effectiveness of Treatment
Depends on Number of microbes

Environment (organic matter, temperature, biofilms)

Time of exposure Microbial characteristics

Actions
Alteration of membrane permeability Damage to proteins

Damage to nucleic acids

Standard Precautions
1. Medical asepsis Wash hands Sanitize & disinfect equipment 2. Use of PPE Gloves, Mask, Gown 3. Deal with waste & disposal OSHA --- deals with waste in facility EPA --- deals with waste outside facility 4. Sterilize instruments 5. Spillage

Handwashing

14

Using alcohol based hand rub

15

SPILLS
should be promptly cleaned up clean up = * gloved employee * paper towels * infectious waste container after clean up :
disinfect area with bleach solution
household bleach = 5.25% sodium hypochlorite bleach solution = 1:10 dilution

Surgical Asepsis
Surgical Asepsis and Sterilization
Steam Sterilization (Autoclave)

Dry Heat Sterilization

Gas Sterilization Chemical Sterilization

1. Chemical agents

Alcohols ,Chlorine, Formalin Suitable for skin and instruments Ultraviolet and Ionizing Radiation
Suitable for sterile room and inoculation hood

2. Radiation

3. Dry Heat

Direct flaming: e.g. inoculating loop Hot-air sterilization :160 , 2h in hot air oven 12130min in autoclave Suitable for medium and instruments
Removal of bacteria by filter medium
Used for heat sensitive materials and filtrated air

4. Moist Heat

5. Filtration

Heat sterilization how it works?

Dry heat- protein oxidation Moist heat- protein denaturation

Wrapping for Sterilization Techniques

involves sanitizing (washing & cleaning) make sure things are in proper working order these materials allow steam or chemical vapors to penetrate, but do not allow airborne or surface contaminants to enter muslin = cotton fabric----------------Shelf Life = 1 month special paper ------------------------- Shelf Life = 1 month special disposable envelopes ------ Shelf Life = 3 months Technique for paper & muslin : Diamond; Bottom; Right Side; Left Side ; Fold Up

Dry Heat
Flaming Hot air oven -170 C for 1 hour -140 C for 3 hours

Moist Heat
1. Pasteurization( below 100 C)
Destroys pathogens without altering the

flavor of the food.

Low temperature (holding method): 63 , 30 min High temperature (flash method) : 72 , 20sec

2. Boiling (at 100 C)

-killing most vegetative forms of bacteria

-10 min or longer time 3. Autoclaving (above 100 C) -killing both vegetative organisms and endospores -121-132 oC for 15 min or longer

Moist Heat vs Dry Heat

Moist heat
Penetrating potency Temp. for protein clotting Extra heat released from condensation higher lower yes

Dry heat
lower higher no

Sterilizing potency: Moist heat Dry heat

Why is moist heat more efficient than dry heat ?

Conductivity. Moisture conducts the heat better than a

dry system.

Sterilization processes
Batch sterilization Continuous sterilization

Filtration
Sterilize solutions that may be damaged or denatured by high temperatures or chemical agents.

The pore size for filtering bacteria, yeasts, and

fungi is in the range of 0.22-0.45 m (filtration

membranes are most popular for this purpose).

Air Sterilization
Very large volumes of sterile air is required in many aerobic

fermentation process.

Methods for air sterilization

Radiation

High temperature

Electrostatic bacteria removal

Filtration

WASTES
1. Medical 2. Contaminated 3. Non-Medical

Medical ( Infectious) Waste

1. laboratory waste = chemicals, detergents, containers 2. pathology specimens = human tissue, blood, excrement, secretions 3. surgical specimens = human parts removed at surgery / autopsy 4. sharps = any device having rigid areas capable of cutting / piercing 5. liquid body waste = stool, excretions, secretions 6. contaminated waste things that have been used on body (dressings)

Non-Medical Waste
- non-patient care waste - kitchen waste

Aseptic Technique
1. Remove all jewelry (e.g., watches, rings, bracelets, necklaces.

Aseptic Technique
2. Put on non-shedding coats, gowns, or coveralls (hospital scrubs); head and facial hair covers; face masks; and shoe covers.

Note that it is important to follow the sequence of items indicated in this step.

Personnel protective equipment

Keyboard and mouse with proper barrier protection

Aseptic Technique
3. Scrub hands and arms to the elbows thoroughly with an antiseptic cleanser (e.g., povidone-iodine or chlorhexidine gluconate).

Aseptic Technique
4. Clean the laminar flow hood with isopropyl alcohol. The alcohol must remain in contact with the surface for 30 seconds prior to compounding any sterile product.

Aseptic Technique
5. Place only essential materials under the airflow hood no paper, pens, or labels. Remove the selected syringe(s) from its overwrap, attach a needle, then discard the waste.

Aseptic Technique
6. Scrub again and glove.

Aseptic Technique
7. Swab or spray needle-penetration closures on vials, injection ports, and other materials.

Aseptic Technique
8. Prepare the sterile product by withdrawing the medication from vials or ampules introducing it into the IV container.

Aseptic Technique
9. Complete a quality check of the product for container integrity and leaks, solution cloudiness, particulates, color of solution

proper preparation of product.

Figure 10.2

Handling Issues
Safety Note!

Coughing and talking should be directed away from the hood.

Coughing -About 3,000 droplets - up to 50 miles per hour. Sneeze - 40,000 more than 200 miles per hour.

HANDLING AND DISPOSAL OF HAZARDOUS AGENTS

Receipt and Storage of Hazardous Agents:
Hazardous drugs should be delivered directly to the storage area, inventoried, and, if necessary, refrigerated. Access to storage areas and work areas for hazardous materials should be limited to specified trained personnel.

S# Waste class 1.

Type of container Colour Yellow

Plastic Human anatomical waste, Animal waste Solid (biomedical waste) Plastic Microbiology and Biotechnology waste Waste sharp Plastic bag puncture proof containers

2. 3.

Red Blue/White Translucent Black

4.

Discarded medicines Plastic bag and Cytotoxic waste Chemical waste (solid) Incineration waste

Reporting Guidelines
National Notifiable Disease Surveillance System Examples HIV/AIDS Cholera Rubella Hepatitis Legionellosis Lyme disease Mumps Smallpox Tetanus Tuberculosis

Sterilization of Surgical Instruments

1933

Steam Sterilization

1947

1906

Steam Sterilization (1974)

Microwave Oven (1947)

Early type of hot air sterilizer

Advanced Sterilizer
Plasma sterilizing system (1993)

Low temperature system for endoscopic devices (1989)

Steam Sterilization
Steam must contact items surface

Filtration

HEPA removes microbes >0.3 m Membrane filtration removes microbes >0.22 m

Physical Methods of Microbial Control

Low temperature inhibits microbial growth Refrigeration Deep-freezing Lyophilization High pressure denatures proteins Desiccation prevents metabolism Osmotic pressure causes plasmolysis

Radiation

Figure 7.5

Radiation
Ionizing radiation (X rays, gamma rays, electron beams) Ionizes water to release OH Damages DNA Nonionizing radiation (UV, 260 nm) Damages DNA

Microwaves kill by heat; not especially antimicrobial

Principles of Effective Disinfection

Concentration of disinfectant Organic matter

pH
Time

Biguanides
Chlorhexidine
Disrupt plasma membranes

Halogens
Iodine
Tinctures: In aqueous alcohol Iodophors: In organic molecules Alter protein synthesis and membranes

Chlorine
Bleach: Hypochlorous acid (HOCl) Chloramine: Chlorine + ammonia Oxidizing agents

Alcohols
Ethanol, isopropanol
Denature proteins, dissolve lipids Require water

Table 7.6

Heavy Metals
Ag, Hg, and Cu
Silver nitrate may be used to prevent gonorrheal ophthalmia neonatorum Silver sulfadiazine used as a topical cream on burns Copper sulfate is an algicide

Oligodynamic action
Denature proteins

Surface-Active Agents, or Surfactants

Soap
Acid-anionic detergents Quarternary ammonium compounds (cationic detergents)

Degerming
Sanitizing

Bactericidal, denature proteins, disrupt plasma membrane

Chemical Food Preservatives

Organic acids
Inhibit metabolism Sorbic acid, benzoic acid, and calcium propionate Control molds and bacteria in foods and cosmetics

Nitrite prevents endospore germination Antibiotics

Nisin and natamycin prevent spoilage of cheese

Aldehydes
Inactivate proteins by cross-linking with functional groups (NH2, OH, COOH, SH) Use: Medical equipment
Glutaraldehyde, formaldehyde, and orthophthalaldehyde

Gaseous Sterilants
Denature proteins Use: Heat-sensitive material
Ethylene oxide

Infrared rays

Temperature =160 Time = 60-120 min

Plasma
Free radicals destroy microbes Use: Tubular instruments

Supercritical Fluids
CO2 with gaseous and liquid properties Use: Medical implants

Peroxygens
Oxidizing agents Use: Contaminated surfaces
O3, H2O2, peracetic acid

Disinfectant

Status

Use

Alcohols (70% or 90%) (intermediatelevel)

Glutaraldehyde (high-level)

bactericidal, tuberculocidal, fungicidal, and virucidal

broad antimicrobial range, fungicidal and virucidal concentrations of 1000 ppm inactivate bacterial spores bactericidal, virucidal, fungicidal, tuberculocidal in 12 minutes at room temperature 6% solutions effective against some bacteria, fungi, and viruses vegetative bactericidal, M. tuberculosis, most viruses and fungi, no sporicidal capability most formulations are tuberculocidal, bactericidal, virucidal, and fungicidal not recommended for high-, intermediate- or low-level disinfection

to disinfect thermometers, medication vials, etc.

to disinfect endoscopes, thermometers, and rubber items to disinfect countertops, floors, other surfaces

Chlorine Compounds (dilution of 1:50 is high-level) Orthophthalaldehyde (high-level)

to clean and process endoscopes

Hydrogen Peroxide (low-level)

may be used to clean work surfaces, not widely used in health care settings

Iodine and Iodophors (intermediatelevel)

may be used as disinfectant or antiseptic

Phenolics (intermediate- or low-level)

have toxic effects, used as environmental not sporicidal disinfectants

Quaternary Ammonium Compounds

cleaning agents for noncritical surfaces

Microbial Characteristics

Figure 7.11

Endospores and Mycobacteria

Table 7.7

Disposal container for contaminated sharps