Repeat the sterilization and streaking 2 more times. Close the Petri plate, and sterilize the inoculation loop. Once inoculated, the broth or agar plate should then be incubated at the ideal growth temperature for the given microorganism to obtain viable culture.
On solid medium, a lawn or continuous strand of bacteria would be visible on agar covered by the first two streaks, but individual colonies should be obtained on the final streak. Poor aseptic techniques would result in the growth of mold and other contaminants on the plate. Aseptic techniques are important in many experiments involving microbial samples from the environment.
In this study, researchers isolated bacteriophages, which are bacteria-infecting viruses, from the common soil bacterium Arthrobacter. Arthrobacter cultures were first grown under aseptic conditions. Soil samples were then washed and filtered in phage buffer, and the phage solution was mixed with the bacterial culture and plated onto agar plates.
A bacterial lawn would form on the plate, but there would be clearings, or "plaques", at spots where the virus had infected and killed the bacteria. Phage could then be purified from these plaques for further study. Other than using Bunsen burners, aseptic working environments can also be maintained in specialized workstations known as laminar flow hoods, which use directed airflow and filters to maintain sterility.
Here, scientists worked in a flow hood to isolate potential pathogenic bacteria and viruses from water samples. These isolates were then cultured together with amoebae. Because amoebae normally eat or "phagocytose" bacteria, any bacteria that were able to resist amoebal digestion and remain in these organisms can also potentially remain viable in human cells and cause diseases. Finally, sterile conditions permit detailed study of ecological mechanisms such as the formation of root nodules in legume plants - bacteria-filled organs that "fix" atmospheric nitrogen into ammonia, which is used by the plant for growth.
Researchers here created "microcosms" for studying the nodulation process using notched Petri plates with plant growth medium, placed seedlings into them and inoculated the seedlings with nodule-forming rhizobial bacteria. The aseptic environment of the flow hood prevents contamination of the cultures with other bacteria or fungi. You've just watched JoVE's video on aseptic techniques in environmental science.
You should now understand why aseptic working conditions are important; how to aseptically perform microbiological experiments; and some applications of aseptic techniques to environmental research. As always, thanks for watching! The outcome of the procedure demonstrates proper aseptic technique and poor aseptic technique. Figure 7 illustrates the contamination that can arise from poor aseptic technique when pouring agarose plates top plate: sterile medium; bottom plates: contaminated media.
Figure 7: Contamination that can arise from poor aseptic technique when pouring agarose plates. Top plate: sterile medium; bottom plates: contaminated media. Proper use of aseptic technique is vital for environmental microbiologists when sampling in the field and in the laboratory when working with media, reagents, and cultured isolates. Poor aseptic technique in the field can result in the transfer of microorganisms from the technician to critical environmental samples, as well as the cross-contamination of microbes from one sample to another.
Such events are of importance, for example, in microbial ecology studies seeking to identify and compare bacterial and fungal populations that may be present in a given biome. Contamination of such samples can result in a loss of data integrity. Aseptic technique is also critical for the maintenance of laboratory culture isolates originating from field sampling or from well-established microbial and cell culture repositories.
Environmental Microbiology. Aseptic Technique in Environmental Science. To learn more about our GDPR policies click here. If you want more info regarding data storage, please contact gdpr jove. Your access has now expired. Provide feedback to your librarian. If you have any questions, please do not hesitate to reach out to our customer success team. Login processing This is a sample clip.
Sign in or start your free trial. Previous Video Next Video. Overview Source: Laboratories of Dr. Charles Gerba - The University of Arizona Demonstrating Author: Luisa Ikner Aseptic technique is a fundamental skill widely practiced in the field of environmental microbiology that requires a balance of mindfulness and practice in the laboratory.
Log in or Start trial to access full content. Preparation for Aseptic Work Obtain and apply the following PPE items: lab coat, latex or nitrile gloves free from tears or holes , and safety goggles Figure 1. For safety in the event of using an open flame, tie back long hair. If you notice that someone fails to wash hands or sterilize equipment, speak up. Doing so may save you or a loved one from potentially fatal infections.
Clean intermittent self-catheterization is recommended when you have a condition that affects your ability to empty your bladder properly. Did you know your kitchen is most likely the dirtiest spot in your home?
Find out where else germs live and how to keep those spots clean. Learn the ins and outs of MRI vs. X-ray imaging tests, including the pros and cons of each test, how they compare to CT scans, how much they cost, and…. Health Conditions Discover Plan Connect. Aseptic Technique. Medically reviewed by Carissa Stephens, R. Uses Types Aseptic vs. What is aseptic technique used for?
Aseptic technique types. Aseptic technique vs. Aseptic technique at home. Aseptic technique benefits. Aseptic technique complications. Aseptic technique outcome. Read this next. What Are Nosocomial Infections? Ideally the equipment should be kept out of direct sunlight as this could also affect the packaging, as well as the quality of the sterile products of the equipment.
It is also sensible to store the equipment in a manner that enables staff to locate it easily, preventing wasted time. Preparation of aseptic technique equipment before performing a procedure should be done in a clean area. A suitable surface should be prepared, such as a dressing trolley or a procedure tray, by cleaning it with a detergent wipe or according to local policy Loveday et al, Ideally, preparation should be undertaken far enough away from hand washbasins to prevent droplets from the sink outlet contaminating the prepared equipment Centers for Disease Control and Prevention, Each sterile package should be inspected to ensure the packaging is still intact with no visible damage; the expiry date must be checked to ensure the equipment is still in date Lloyd Jones, All equipment required should be gathered at this point and laid out in a manner to protect key parts using sterile packets, covers and caps Rowley et al, Alternatively, a sterile field can be created using a sterile dressing pack Lloyd Jones, It is important to inform the patient before undertaking any procedure and to obtain consent, allowing time to explain the specific procedure and to help reduce any anxieties the patient may have Royal College of Nursing, To reduce the risk from airborne dispersal of micro-organisms, the procedure area should be prepared by closing windows, turning off any fans that are in use and avoiding any bed making being undertaken in close proximity Lloyd Jones, A clean disposable apron provides an ideal barrier between potentially contaminated uniforms and the procedure, reducing any contamination that may arise from the procedure.
The decision as to whether single-use sterile or single-use non-sterile examination gloves should be worn, or whether gloves are required at all, should be based on a risk assessment NHMRC, ; Loveday et al, Single-use non-sterile gloves should be worn to protect the healthcare worker when there is a risk of contact with blood or body fluids during any procedure Loveday et al, Where there is a risk that key parts or key sites cannot be protected, for example, during urinary catheter insertion, sterile gloves should be worn NHMRC, ; Rowley et al, During the procedure, the sterile field needs to be maintained; this can be achieved by careful opening of the sterile packets to avoid contamination of the sterile equipment and the sterile surfaces of the inside packaging.
If a sterile dressing pack is being used, care should be taken to ensure that only the corners are used when opening out the sterile field. Carefully open any other sterile equipment and gently place it onto the sterile field, avoiding any contamination of the sterile surfaces of the equipment Lloyd Jones, At the end of the procedure, all waste must be disposed of in the appropriate waste stream bin, such as a healthcare waste bin. Sharps, including needles, suture cutters, scissors and blades must be disposed of at the point of use in an approved sharps container Loveday et al, The remaining waste, including aprons and gloves, should be disposed of according to local policy Loveday et al, When all the waste has been disposed of safely and apron and gloves, if worn, have been removed hands should be decontaminated Loveday et al, Asepsis or aseptic means the absence of germs, such as bacteria, viruses, and other microorganisms that can cause disease.
Healthcare professionals use aseptic technique to protect patients from infection. This article will explain different types of aseptic technique, how it differs from clean technique, and how to use it at home. A person is vulnerable to infection as soon as there is a break in their skin, regardless of whether it occurs as a result of an accidental injury or a surgical incision. Aseptic technique helps prevent healthcare-associated infections HCAIs.
An HCAI is an infection that a person acquires as a result of treatment from a healthcare professional. These types of infection are a significant concern in the healthcare community. HCAIs can lead to severe health complications for affected individuals and disciplinary consequences for medical facilities. Additionally, HCAIs have a significant economic impact.
Aseptic techniques range from simple practices, such as using alcohol to sterilize the skin, to full surgical asepsis, which involves the use of sterile gowns, gloves, and masks. Healthcare professionals use aseptic technique practices in hospitals, surgery rooms, outpatient care clinics, and other healthcare settings. Using aseptic technique prevents the spread of infection by harmful germs. Healthcare professionals use aseptic technique when they are:.
Aseptic technique and clean technique are two closely related healthcare practices that both aim to keep people safe from infection. The aim of using aseptic technique is to eliminate germs, which are disease-causing microorganisms. Clean technique focuses on reducing the number of microorganisms in general. Healthcare professionals learn both aseptic and clean techniques and use them in different situations. Clean technique is often sufficient for long-term care, in-home care, and some outpatient clinical settings.
Healthcare professionals use clean technique for people who are not at high risk of infection.
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