Media preparation, proper storage, and quality control testing can assure a consistent supply of high quality media. To identify sterilized items use indicator tape to label the flask (medium name/code, preparation date, initials). A formal record retention and retrieval program should be in place. Water is the universal diluent for microbiological media. When exposed to moisture, a hard mass is formed, which alters the chemical and microbiological properties of the medium. Congress delegated such authority through the Safe Drinking Water Act (SDWA), which mandates the establishment of a list of contaminants and their corresponding maximum contaminant levels and maximum contaminant level goals. Use sterile, graduated, Close screw caps tightly after autoclaving. User-laboratory quality control tests on prepared media. The frequency of calibration and performance verification will vary based on the type of instrument and the importance of that equipment to the generation of data in the laboratory. Let the media cool down to 45-50C before adding heat-labile supplements, Filter-sterilize heat-labile supplements (not in the case of blood), Let supplements come to room temperature before adding, Verify pH (usually at 25C) with a pH meter before dispensing, Take 20 mL out of the flask, let it cool down, and measure pH. Some of the technologies available to small treatment plants to inactivate organic chemicals, some are already in use, include powdered activated carbon. Use chemical indicators with each cycle (e.g., time-steam-temperature strips) and biological indicators on an interval basis to verify autoclave cycles. Therefore, the level of containment necessary for working safely with bacterial cultures also varies according to a system that classifies microbes into one of four biosafety levels (BSL), which provides minimum standards for safe handling of microbes at each level. Record product name, lot number, preparation date, weight and volume of water, and operator identification in a logbook. In the United States, states are encouraged to develop and run their own hazardous waste program; otherwise, the EPA runs the program for the state. An example of data being processed may be a unique identifier stored in a cookie. As an asst. In the United States, the National Institutes of Health posted a waste disposal guide online in 2014 that directly addresses culture media containing antibiotics.10 The guidance document indicates the following: Before disposal, cell culture media must be decontaminated (see MPW Section for instructions) either by steam autoclave or adding disinfectant directly to vessel or treating pooled spent media. b. their susceptibility to laboratory disinfectants. This web page provides information on how to work with and dispose of bacterial cultures used in the WSSP research project. Facilities should be designed so that raw material and excipient sampling can be done under controlled conditions, including proper gowning and sterilized sampling equipment. Safeguarding the quality of this media is therefore critical to the success of the microbiology laboratory. Autoclaving by moist heat is the preferred sterilization technique, except in instances when boiling is required in order to avoid deterioration of heat-labile components of the media. The primary hazards to people working in a BSL 1 or 2 laboratory relate to. There are options, and some are easier to adopt than others. Also, from a public health perspective, overuse or abuse of antibiotics is known to influence microbial resistance.3. Sodium hypochlorite, the active ingredient in chlorine bleach, is routinely used in the laboratory to decontaminate surfaces and equipment or deactivate biological materials by inactivating vegetative bacteria, fungi, lipid and non-lipid viruses, and other liquid specimens. We and our partners use data for Personalised ads and content, ad and content measurement, audience insights and product development. These barriers include protective clothing, sanitization and disinfection procedures, and biological safety cabinets designated for clean or aseptic operations only. Subculturing, staining, microbial identification, or other investigational operations should be undertaken in the live culture section of the laboratory. Because of these characteristics of microbiological analysis, laboratory studies should be conducted with the utmost care to avoid exogenous contamination as previously discussed in this chapter. https://www.epa.gov/sites/production/files/2016-06/documents/npwdr_complete_table.pdf, http://www.formatex.info/microbiology4/vol3/1455-1460.pdf, https://www.epa.gov/sites/production/files/2015-08/documents/white_paper_aquatic_life_criteria_for_contaminants_of_emerging_concern_part_i_general_challenges_and_recommendations_1.pdf, https://www.epa.gov/sites/production/files/2015-08/documents/sab_advisory_on_aquatic_life_wqc_for_contaminants_of_emerging_concern.pdf, https://www.epa.gov/fedfac/emerging-contaminants-and-federal-facility-contaminants-concern, https://www.federalregister.gov/documents/2009/10/08/E9-24287/drinking-water-contaminant-candidate-list-3-final, https://www.orf.od.nih.gov/EnvironmentalProtection/WasteDisposal/Documents/Final%202014%20WASTE%20GUIDE%20PRINT.pdf, http://sti.epfl.ch/files/content/sites/sti/files/shared/security/dechets-bio-1-2.pdf, http://www.su.se/polopoly_fs/1.236898.1432209877!/menu/standard/file/procedures_disposal%20of%20liquid_150323.pdf, http://science.gu.se/digitalAssets/1383/1383230_guidelines-for-wastewater-from-laboratory-research-and-teaching_120928.pdf, http://static.usp.org/pdf/EN/referenceStandards/msds/1709007.pdf, https://tools.thermofisher.com/content/sfs/manuals/zeocin_pps.pdf, Beta-lactams: ampicillin, carbenicillin, penicillin. Technical Support - FAQs Read more: Automated Culture Media Preparation and Dispenser System. All articles and SOPs are written by Ankur Choudhary. Here's how. In tissue culture media traps change at least every 3 months (indicate the date of the last change on the flask). Culture Media - Microbiology Resource Center - Truckee Meadows Which of the following is required when working with BSL 2 agents? Run an empty Autoclave sterilization cycle at 15 lbs for 15 minutes after the sterilization of used media. . Ready to use SOPs, Protocols, Master Plans, Manuals and more Worldwide Regulatory Updates Pharmaceutical News Updates Interview Questions and Answers All Guidelines in One Place. Ideally, staff assigned to sampling activities, particularly those in support of aseptic processing, should not work in the vicinity of live culture laboratory operations. If contamination risk: keep lids closed. Within this list, contaminants are classified as disinfectant, disinfectant by-product, inorganic chemical, microorganism, organic chemical, or radionuclide, but antibiotics are not listed as contaminants. Where XXX indicates the year in which activity is performed e.g. Clin Microbiol Infect. a . They should not independently conduct a microbial test until they are qualified to run the test. Sterilization of media should be performed within the parameters provided by the manufacturer or validated by the user. Equipment 4.1 Maintenance of equipment 4.2 Quali cation 4.3 Calibration, performance veri cation and monitoring of use 5. . Corresponding Author: Esmeralda L. Meyer, Environmental Health and Safety Office, Emory University, Atlanta, GA 30322, USA. If in doubt as to how to handle or dispose of any specific medium consult the local waste disposal authority. In case of deviation, verify water, dehydrated medium, glassware, and procedure and start again. Microbiologists culture bacteria by providing them with food, water, and other growth requirements in an environment with a constant and comfortable growth temperature. Ready-to-use cultures may require confirmation of purity, identity, and inoculum size. ), the volumes of culture produced (is the pathogen being propagated? Media prepared in-house should be stored under validated conditions. In addition, the growth characteristics of the microorganism should be considered (especially in questions of the growth of filimentous fungi in liquid media). Validation of test methods 4. If your lab can afford to purchase them, automated culture media preparation systems can save you manual labor and time. The over-riding principle is that the test should be performed as written in the SOP, the SOP should be written to reflect how the test is actually performed, and the laboratory notebook should provide a record of all critical details needed to confirm the integrity of the data. A set of techniques, mostly developed in the late 19th century by Robert Koch, Louis Pasteur, and their collaborators, permitted the isolation of bacteria from their natural environments and separation into pure cultures for further study. Typically, manufacturers recommend using an autoclave cycle of 121, The pH of each batch of medium should be confirmed after it has cooled to room temperature (25. The performance of media prepared in a laboratory or by a manufacturer is highly dependent on preparation. Use a standardized and validated rack. Run an empty Autoclave sterilization cycle at 15 lbs for 15 minutes after the sterilization of used media. x 4 in.) Improper media preparation can cause unsatisfactory conditions for microbial growth or recovery and unreliable results. If possible, any sample found to contain growing colonies should not be opened in the clean zone of the laboratory. Following the approval and implementation of the corrective action plan, the situation should be carefully monitored and the adequacy of the corrective action determined. __________________________________________________________________________, 2. Dehydrated media are stored in a cool, dry place, protected from light and dust. Safety glasses or goggles are also recommended. Defined media and complex media are two broad classes of culture media used in microbiology. While there are no specific requirements in the United States for the management of antibiotics and other pharmaceuticals in cell culture media generated in the laboratory, it could be considered a best practice to start the conversation in your institution. Stir or rotate for a few minutes (do not shake! The summary provided here is just the beginning of the conversation. All of the equipment and supplies used in experiments involving bacterial cultures should be sterilized. a. accidental skin punctures caused by mishandling of sharp lab tools. For disposal of used culture media, follow in-country or WHO guidelines, which recommend inactivation and subsequent incineration before disposal. Therefore, record the opening date on the container and use the product for a maximum of 6 months after opening unless otherwise specified by the manufacturer. b. It is critical to know if the finding is statistically significant in light of assay variability. Precautions in the use and disposal of prepared media. Disposal Of Microbial Cultures And Culture Media In addition, specific laboratory rules must be followed for containment of microbial cultures in the laboratory, for the safety of all. This will prevent extraneous contamination from being carried into controlled environments and will prevent false-positive results. We and our partners use cookies to Store and/or access information on a device. Table 1 has been updated from the original posting at the University of Gothenburgs website under the sections Rules for Handling Antibiotics and Handling Instructions to add a few more antibiotics commonly used in culture media.13 The recommendation provided here is for antibiotics contained in media; it should not be used for disposal of stock solutions. Tests routinely performed on in-house prepared media are pH, growth promotion, and periodic stability checks to confirm the expiry dating. It is essential that cross-contamination of microbial cultures be minimized to the greatest extent possible, and it is also important that microbiological samples be handled in an environment that makes contamination highly unlikely. Importantly, it reiterated that bleach not be added to cell culture media that requires inactivation by heat.11, The University of Stockholm published an online document titled Procedures for the Disposal of Liquid Chemical Residues and Aqueous Solutions. This guidance document covers laboratory research and teaching laboratories within the university that are connected to a very specific part of the sewage system in the city of Stockholm. Then carefully remove the saturated paper towels, dispose of them in the biohazard waste, and clean the area again with disinfectant. Dispensing at too high temperature leads to excessive evaporation. Hello, thank you for visiting my blog. Recognize the international symbol for biohazards, and know where and how to dispose of all waste materials, particularly biohazard waste. If other ingredients are to be added (e.g., supplements such as sheep blood or specific vitamins, nutrients, growth promoters, or antibiotics), they should be incorporated when the molten agar has cooled, just before distribution to plates. 2023 Mary Ann Liebert, Inc., publishers. Agar plates, like a lot of lab refuse and equipment, will need to be sterilized before either disposal (for polystyrene plates) or reuse (for glass plates).Sterilizing polystyrene agar plates before disposal follows . Inhibitory substances can come from detergent residue after cleaning glassware or from prior materials used in the glassware. Disposal of used culture media. Generally, good microbiological practices recommend that the biohazard be treated before disposal of the culture . Institutions generating hazardous waste must meet the requirements of the state or federal plan. Decontaminated media must be collected as chemical waste and called for pick up by Chemical Waste Services. Allow the oven to cool to 50C before opening (to avoid cracking glassware).