Aseptic Technique Lab Report

Aseptic Technique Marlene P. Tubieros Introduction Bacteria are everywhere, and some are good for us while others are harmful. That’s why minimizing our exposure to harmful bacteria when undergoing medical procedures is so important. Healthcare providers regularly use aseptic techniques to achieve this. Aseptic technique is a method designed to prevent contamination from microorganisms. It involves applying the strictest rules and utilizing what is known about infection prevention to minimize the risks that you’ll experience an infection. Objectives   

To acquire the skill of aseptic technique in the field of Microbiology. To prevent contamination of cultures and media from microbes in the environment. To transfer cultures from one medium by inoculating another medium.

Materials and Methods 1. Using a marking pen, label the test tubes correctly. 2. Sterilize your inoculating loop by passing it at an angle through the flame of the alcohol lamp until the entire length of the wire becomes glowing red/orange from the heat. (NOTE: Never lay the loop down once it is sterilized or it may become re-contaminated.) 3. Using your other hand, pick up the tube containing the culture and gently shake it to disperse the culture. 4. Loosen the cotton plug of the test tube which contains your bacteria culture with the little finger curled towards the palm of your right hand. 5. Do not put down the cotton plug. 6. Flame the neck of the test tube of bacteria culture by passing the neck forwards and back through a hot alcohol lamp flame. 7. Using the inoculating loop, get an ample amount of culture in the test tube. Carry out the transfer of cultures as quickly as possible, with tubes open to the air for the minimum length of time. (NOTE: Work close to the alcohol lamp flame to reduce the chances of contamination.) 8. Again, flame the neck of the test tube that contain your cultured bacteria and return the cotton plug using your little finger. 9. Get the test tube of sterile nutrient broth. Carefully remove the cotton plug) with your little finger of the hand holding the inoculating loop. 10. Flame the neck of the test tube of sterile nutrient broth. 11. Insert the inoculating loop into the sterile broth and inoculate it by gently moving the loop back and forth in order to disperse the cells.

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Remove the inoculating loop from the test tube and flame the mouth of the tube again then replace the cotton plug.

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Sterilize the loop by flaming it. Now you may lay it down on the lab bench or return it to its container.

Result and Discussion Aseptic technique aims to prevent pathogenic organisms, in sufficient quantity to cause infection, from being introduced to susceptible body sites by the hands of staff, surfaces or equipment. It protects patients during invasive clinical procedures by utilising infection prevention measures that minimise the presence of micro-organisms. Aseptic transfer is transferring living microbes from one place to another without contamination of the culture, the sterile medium, or surroundings. In addition, aseptic technique is of utmost importance to maintain pure stock cultures while transferring cultures to new media. Aseptic technique is also essential for isolation of a single species of microorganism from a mixed culture to obtain a pure culture. Furthermore, proper aseptic technique prevents microbes used in the laboratory from accidentally being released into the environment and/ or infecting people working in the laboratory. This is especially relevant when pathogens are being handled 1. Explain why the following steps are essential during subculturing. A. Flaming the inoculating prior to and after each inoculation. 

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Holding the loop in the flame of the Bunsen burner kills all contaminating organisms, thus sterilizing the loop Inoculation loops need to be sterilised before and after every use to reduce the chances of contamination as much as possible to achieve the most accurate results possible within tests.

B. Cooling the inoculating instrument prior to obtaining the sample 

If the inoculating instrument is too hot it may kill the bacteria you are trying to obtain on contact

Conclusion Aseptic technique is a fundamental and important laboratory skill in the field of microbiology. Microbiologists use aseptic technique for a variety of procedures such as transferring cultures, inoculating media, isolation of pure cultures, and for performing microbiological tests. Proper aseptic technique prevents contamination of cultures from foreign

bacteria inherent in the environment. For example, airborne microorganisms (including fungi), microbes picked up from the researcher’s body, the lab bench-top or other surfaces, microbes found in dust, as well as microbes found on unsterilized glassware and equipment, etc. may potentially contaminate cultures, thus interfering with the lab results. Using proper aseptic technique can greatly minimize or even eliminate the risk of contamination. References http://www.biotopics.co.uk/microbes/tech1.html http://www.austincc.edu/microbugz/aseptic_technique.php