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Microbiology, Lecture 18+19 Prt1

Microbiology, Lecture 18+19 Prt1

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Published by: Mohamed Harun B. Sanoh on Apr 28, 2011
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02/08/2013

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Today we're going to talk 
about Sterilization, Disinfection, & Microbial Waste Disposal
,which is the main topic of this lecture. And about the first part
of non specific immunity
.So lets begin with some terms:
 Sterilization
is the complete destruction of all microbes, including cells, spores, and viruses.And can be accomplished by physical methods such as radiation, heat. And can beaccomplished by chemical methods.In contrast to sterilization,
disinfection is
destruction or removal of all or most pathogensfrom objects by physical or chemical methods.
So sterilization is the destruction of all microbes, pathogenic or non pathogenic. Whiledisinfection only relates to pathogenic microorganisms
.An example of disinfection technique is basically wiping your skin with alcohol. It willremove most of microorganisms of it.Another example of disinfection is the process of  pasteurization of liquids such as milk. So you heat the milk for a certain temperature for several times, and you do that to remove mainly brucella. We're not going to talk aboutanother micro organisms here, only brucella.So pasteurization, at the bottom line, is a type of disinfection technique.
Disinfectants
are chemicals that we use to perform regular disinfections and usuallydisinfectants are used for non living objects such as bench tops.Disinfectants usually can killmost vegetative bacteria, most growing dividing bacteria, but cannot destroy regular sporesor bacterial spores.
Antiseptics
, these are basically disinfectants that are safe enough to be used on living tissues.So 70% alcohol is a type of antiseptics, whereas bleach is a disinfectant because it's veryharmful for the skin, and if u put it on metals it might ruin the metals.Whenever you see a word that ends with –cide or –cidal, that basically means death so agermicide agent is an agent that kills germs.
Bactericidal agent
is an agent that kills bacteria, but not necessarily bacterial spores.A
sporocidal agent
is an agent that kills bacterial spores.
A fungicidal agent
is an agent that kills fungi including their spores.
And Algicidal agents
is an agent that kills algae.
A viricidal
agent destroys viruses.We took this term for microbistatic, and
microbistatic agent
is an agent that inhibits growthand metabolism of bacteria, but not necessarily kill them. So if you remove the microbistaticagent from a microorganism, this microorganism can resume its growth.This is obviously a contrast
to bactericidal agent
that will actually kill the bacteria.Final two terms are sepsis and antisepsis.
Sepsis r
efers to the presence of pathogen in blood or tissues, whereas
asepsis (antisepsis
)means the absence of pathogens.
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Whenever you try to prevent an infection, the procedure or whatever you are trying to do isreferred to as antisepsis. So cleaning of surfaces using disinfectants, is relate to antisepsistechnique.
 Now let's discuss the various types of physical and chemical methods that are used to inhibit bacterial growth. And we'll begin with the
 physical methods
.
Basically we have the heat method, which is the most common method used to inhibit bacterial growth or destroy microorganisms, and we have two main types.1)Dry heat. Basically increase the temperature without adding moisture such as in thecase of the dry hot oven, or in the case of directly using the flame or a heating deviceto burn the organism. So later on as dentists, you use the oven to basically sterilizeyour various equipments. In the oven you place the items at 160 degrees centigrade or you can use higher temperature with less time.2)The second type of heat is moist heat. Examples include boiling and using anautoclave.We have two factors that determine the effectiveness of heat in killing the microbes.1)Temperature.2)The exposure time.So the higher the temperature is, the more effectively sterilization occurs. And the longer thetime of exposure is, the more effectively sterilization occurs.And each microbe has a particular property regarding its sensitivity to heat and this is
calledthe thermal death point.
So the thermal death point of any species is the lowest temperature that will kill all of theorganisms in a standardized pure culture within a specified time. For example, we can seethat bacteria x needs to be exposed to 70 degrees centigrade for 30 minutes in order to becompletely killed. If exposed to 65 degrees for 30 minutes, that will not ensure the killing of all of the bacteria. Obviously if we go above 70 we will kill it. As a minimum you need 70degrees. So 70 degrees for 30 minutes is the thermal death point. And each microbe will haveits own different thermal death point.You probably have seen this in the lab, whenever we are trying to transfer the bacteria fromone place to another, we use an instrument called a loop so the bacteria is attached to one endof the loop, and we use it to streak another plate. And we probably use the flame to sterilizethe loop to prevent contamination between different cultures.We can use another method other than flame such as electrical heating device, which heatsthe tip of the loop to a very high temperature.Autoclave is another example of using heat to sterilize materials. The autoclave is simply alarge metal pressure cooker. So it’s a sealed container that contains some liquids and water.We heat the container and the water starts boiling but since it's under pressure the boiling
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temperature will reach 121 degrees centigrade. And the internal pressure of the water vapor will reach 15 pounds per square inch, or 15 psi.So that high temperature will kill many bacteria and that high pressure that is generated will push the vapor very deep within the material insuring proper sterilization of the bacteria. Soyou can use an autoclave at that temperature of 121 degrees and at pressure 15 psi for at least15 minutes and you insure killing of all microbes that are present in the sample.
 And youhave to memorize these numbers (121 at 15 psi for 15 minutes)
. If you go longer obviouslythat’s fine but if less than any of those numbers you cannot insure the killing of all themicrobes present.A simple example of an autoclave is a pressure cooker and we have also other professionalmachines that can perform autoclaving and they can take a very large volume. And they havecomputers system to control the pressure, temperature and duration of autoclaving.In order to make sure that you have properly autoclaved and sterilized the material, you canadd some type of indicator with the material you try to autoclave in order to see if thetemperature and pressure were reached.Examples of these indicators are specific type of tape or specific types of vials .Here's an example of a very large autoclave (slide #8) so basically you place the materialinside and you close the lid and press start and the machine itself will heat the material andinsure that the heat and pressure were reached, and we'll run the machine for a specific time(at least 15 minutes).Here's an example of the autoclave tape (slide #9) so you sterilize the cloth and you add thisindicator tape, when you started the tape was completely clear, it did not contain these black stripes but exposure to the high heat and the high temperature and pressure of the autoclavewill change these white stripes into black. So the black stripes indicate that the autoclavingwas successful.Another method which we can use is basically introducing these vials which contain bacterialspores that are highly resistant to heat. So we place the vial in the autoclave, and after theautoclave we take it out and incubate it at 37 degrees, which would allow any survivingspores to germinate and grow, and the metabolism of the microorganisms will change thecolor of the tube from purple to yellow. So if you did the autoclave and incubated the tube(vial) at 37 degrees and the color stayed the same that means that you've killed all themicroorganisms in that sample. But, if the tube changes from violet to yellow, that meansthat the autoclaving was not successful.Other physical methods that inhibit bacterial growth;1)Cooling, or freezing the sample will not kill all the bacteria in the sample, in fact it will preserve the bacteria. So once the sample returns to the room temperature or 37degrees, it will resume its growth.
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