4/7/2013

M265 Medical Microbiology

Sterilization, Disinfection, & Microbial Waste Disposal Ref: Burtons Microbiology for the Health Sciences, 9th Ed P131-139,p211,216, Handout

Lecture # 18

Dr. Samer Swedan

Definition of Terms
Sterilization is the complete destruction of all microbes, including cells, spores, and viruses. Accomplished by dry heat, autoclaving (steam under pressure), gas, chemicals, and certain types of radiation. Disinfection is the destruction or removal of all or most pathogens from objects by physical or chemical methods; pasteurization is an example of a disinfection technique. Disinfectants are chemical substances that eliminate pathogens on inanimate objects; they cant kill bacterial spores Antiseptics are solutions used to disinfect skin and other living tissues.

4/7/2013

Definition of Terms, cont.

The suffix cide or cidal refers to killing.
Germicidal agents, biocidal agents, and microbicidal agents are chemicals that kill microbes. Bactericidal agents are chemicals that specifically kill bacteria, but not necessarily bacterial endospores. Sporicidal agents kill bacterial endospores.

Fungicidal agents kill fungi, including fungal spores.

Algicidal agents kill algae. Viricidal (or virucidal) agents destroy viruses.

Definition of Terms (cont.) A microbistatic agent is a drug or chemical that inhibits growth and reproduction of microbes. A bacteriostatic agent is one that specifically inhibits the metabolism and reproduction of bacteria. Sepsis refers to the presence of pathogens in blood or tissues, whereas asepsis means the absence of pathogens. Antisepsis is the prevention of infection.

4/7/2013

Using Physical Methods to Inhibit Microbial Growth

Heat 2 factors temperature and time - determine the effectiveness of heat for sterilization. The thermal death point (TDP) of any species is the lowest temperature that will kill all of the organisms in a standardized pure culture within a specified time. The thermal death time (TDT) is the length of time necessary to sterilize a pure culture at a specified temperature Types of Heat Dry heat e.g., oven (160C for 2h), electrical incinerator, or flame Moist heat boiling or use of an autoclave

Dry Heat Sterilization

Using a Bunsen burner flame

Using an electrical heating device

4/7/2013

Using Physical Methods to Inhibit Microbial Growth, cont.

The autoclave A large metal pressure cooker that uses steam under pressure to completely destroy all microbial life. Increased pressure raises the temperature above the temperature of boiling water (above 100oC) and forces steam into materials being sterilized. Autoclaving at a pressure of 15 psi at 121oC for 15 minutes destroys vegetative microorganisms, bacterial endospores, and viruses. Pressure-sensitive tape or spore strips or solutions Can be used as a quality control measure to ensure proper autoclaving.

A large autoclave.

4/7/2013

Pressure-sensitive autoclave tape showing dark stripes after sterilization.

Copyright 2011 Wolters Kluwer Health | Lippincott Williams & Wilkins

Required Self-Reading Biological Indicators for Monitoring the Effectiveness of Steam Sterilization
Sealed ampules containing bacterial spores suspended in a growth medium are placed in the load to be autoclaved/sterilized. Following sterilization, the ampules are incubated at 37C. If the spores were killed. There will be no change in color of the medium; it will remain purple. If the spores were not killed, acid production by the organisms will cause the medium to change from purple to yellow.

4/7/2013

Using Physical Methods to Inhibit Microbial Growth, cont.

Cold; most microorganisms are not killed, but their metabolic activities are slowed. Desiccation; many dried microorganisms remain viable, but they cannot reproduce. Radiation; UV rays, gamma rays, or beta rays. Ultrasonic waves; used in hospitals and medical and dental clinics to clean equipment. Filters; used to separate cells/microbes from liquids or gases. Gaseous atmosphere; can be altered to inhibit growth.

Using Chemical Agents to Inhibit Microbial Growth

Chemical disinfection refers to the use of chemical agents to inhibit the growth of pathogens, either temporarily or permanently. Disinfectants are affected by: Prior cleaning of the object or surface The organic load (e.g., feces, blood, pus) The bioburden; types and numbers of microbes Concentration of the disinfectant Contact time Physical nature of the object being disinfected Temperature and pH

4/7/2013

Using Chemical Agents to Inhibit Microbial Growth, cont.

Characteristics of an ideal chemical antimicrobial agent: Should have a broad antimicrobial spectrum Fast acting Not affected by the presence of organic matter Nontoxic to human tissues and noncorrosive Should leave a residual antimicrobial film on surface Soluble in water and easy to apply Inexpensive and easy to prepare Stable as both a concentrate and a working solution Odorless

4/7/2013

Using Chemical Agents to Inhibit Microbial Growth (cont.)

Antiseptics May safely be used on human tissues. Reduce the number of organisms on the surface of the skin; do not penetrate pores and hair follicles. Antiseptic soaps and scrubbing are used by healthcare personnel to remove organisms lodged in pores or folds of the skin.

Controversies Relating to the Use of Antimicrobial Agents in Animal Feed and Household Products 40% of the antibiotics manufactured in the U.S. are used in animal feed; microorganisms resistant to these antibiotics survive! Drug resistant organisms are transmitted in animal feces and in food products. Efforts are underway to eliminate or reduce the practice of adding antibiotics to animal feed.

SKIP

Use of antimicrobial agents is widespread in toys, cutting boards, in hand soaps, and many other household products; resistant microorganisms survive! Controversy: Should children be exposed to all sorts of microorganisms for their immune systems to develop properly?

4/7/2013

Infectious Medical Waste

Infectious Medical Waste is medical waste which is capable of producing an infectious disease. Plan: Sterilize all bacterial, fungal, viral and parasitic organisms and cultured cells.

Infectious Medical Waste

Medical waste shall be considered capable of producing an infectious disease if (1) it has been, or is likely to have been, contaminated by an organism likely to be pathogenic to healthy humans, (2) if such organism is not routinely and freely available in the community, AND

(3) such organism has a significant probability of being present in sufficient quantities and with sufficient virulence to transmit disease.

4/7/2013

TYPES OF INFECTIOUS WASTE

Cultures of microorganisms Human blood and blood products Pathological wastes All Sharps (even if not contaminated) Contaminated animal carcasses, body parts, bedding and related wastes Materials (soil, water, or other debris) which result from the cleanup of a spill of any infectious medical waste. Waste contaminated by or mixed with infectious medical waste.

Cultures and stocks of microorganisms

10

4/7/2013

Human Blood & Blood Products

All human blood (wet or dried) Products from human blood.

Pathological Waste
Human pathological wastes tissues, organs, body parts, containers of body fluids

11

4/7/2013

Sharps
Any article that can puncture or cut, and have been used in animal/human patient care or treatment Examples: needles, syringes, scalpel blades, razors, forceps

Disposal of Sharps
Sharps should be handled and disposed of properly Dispose of sharps in specifically designed puncture-resistant containers (sharps containers)

12

4/7/2013

Animal Waste
Contaminated animal carcasses, body parts, animal bedding known to have been exposed to infectious agents during research

PACKAGING DISPOSABLE INFECTIOUS WASTE

1. ORANGE BIOHAZARD BAGS MUST BE USED BY ALL LABS FOR DISPOSABLE WASTE.
2. ALL ORANGE AUTOCLAVE BAGS MUST BE DOUBLE-BAGGED. 3. DO NOT FILL THEM MORE THAN 2/3 FULL! 4. DO NOT TIE THE BAGS CLOSED. Gather the top and place a strip of autoclave tape around the top. Steam must be able to enter and exit the bag. 5. ALL bags and flasks must be LABELED with name, room number, phone number and contents.

13

4/7/2013

STERILIZATION OF INFECTIOUS WASTE

All infectious waste must be autoclaved Bags are then transferred to black plastic bags in a separate location. Disposable waste is taken to the dumpster.

DONT PUT NONINFECTIOUS ITEMS IN ORANGE BAGS

Bubble wrap Paper towels, either from drying your hands after washing them or from wiping down the bench with a disinfectant Scalpel blade wrappers Needle wrappers Packaging materials such as cardboard, Styrofoam, etc. Paper Food wrappers Soda cans!

14

4/7/2013

BLOOD AND BLOODBORNE PATHOGEN RULE

Bloodborne pathogens are organisms carried in human blood that cause disease, including bacteria and viruses like HBV and HIV An Exposure Control Plan is required.

BLOOD AND BLOODBORNE PATHOGEN RULE

Universal Precautions: Treat all human blood and byproducts as if it is contaminated No eating, drinking, smoking, applying cosmetics or contact lenses in the labs No mouth pipetting; use pipetting device Proper disposal (bleach and autoclave)

Protective clothing appropriate to the level of risk and use of barriers to exposure

END OF 2nd EXAM MATERIAL

15