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CULTURE MEDIA Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Culture Media Culture media are mediums that provide essential nutrients and minerals to support the growth of microorganisms in the laboratory. Microbes that grow and multiply in or on a cu...

CULTURE MEDIA Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Culture Media Culture media are mediums that provide essential nutrients and minerals to support the growth of microorganisms in the laboratory. Microbes that grow and multiply in or on a culture medium are known as a culture. Agar is a common solidifying agent for a culture medium. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya A. Classification of culture media based on consistency 1. Solid Media In these media, the agar which is an unbranched long chain of polysaccharides is added in the concentration of 1.5-2.0%. Most commonly, 1.3% agar is used to prepare solid media in labs. The agar-containing media solidifies at 37 ºC. Solid media are used to grow microorganisms in their full physical form, prepare bacterial pure cultures, or isolate bacteria to study colony characteristics. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya A. Classification of culture media based on consistency 2. Semi-Solid Media This media has 0.2-0.5% agar concentration, and due to the reduced agar concentration, it appears as a soft, jelly-like substance. It’s mainly used to study the motility of microorganisms, distinguish between motile and non-motile bacterial strains and cultivate microaerophilic bacteria – bacteria on this media appear as a thick line. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya A. Classification of culture media based on consistency 3. Liquid Media These media do not contain any traces of solidifying agents, such as agar or gelatin, and large growth of bacterial colonies can be observed in the media. Liquid media are also called broths, they allow for uniform and turbid growth of bacterial strains when incubated at 37ºC for 24hrs. The media is used for the profuse growth of microorganisms and fermentation studies. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya A. Classification of culture media based on consistency 4. Biphasic Media Consist of both solid and liquid media. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya B. Classification based on the nutritional component 1. Simple Media It’s a general-purpose media that supports the growth of non- fastidious microbes, and it is primarily used for the isolation of microorganisms. Examples are nutrient broth, peptone water, and nutrient agar. 2. Complex Media These are media containing nutrients in unknown quantities that are added to bring about a particular characteristic of a microbial strain. Examples are tryptic soy broth, blood agar, and nutrient broth. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya B. Classification based on the nutritional component 3. Synthetic media Synthetic media is a type of chemically defined media and is produced from pure chemical substances. A defined media refers to a medium having a known concentration of ingredients, like sugar (glucose or glycerol) and nitrogen source (such as ammonium salt or nitrate as inorganic nitrogen). It is generally used in scientific research, Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 1. Basal media These are routinely used simple media having carbon and nitrogen sources that boost the growth of many microorganisms. They are also known as general-purpose media and are considered non-selective media. The basal media do not require enrichment sources for the growth of non-fastidious bacteria and are suitable for growing Staphylococcus and Enterobacteriaceae. They are generally used to isolate microorganisms in labs or in sub-culturing processes. Examples are nutrient broth, nutrient agar, and peptone water. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 2. Enriched media This media is prepared by adding additional substances like blood, serum, or egg yolk in the basal medium. It’s used to grow fastidious microorganisms as they require additional nutrients and growth-promoting substances. Examples are chocolate agar, blood agar, and Loeffler’s serum slope. Chocolate media is used to grow N. gonorrhea while blood agar (which is prepared by adding 5-10% blood by volume to a blood agar base) is used to identify hemolytic bacteria. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 3. Selective media This media allows the growth of certain microbes while inhibiting the growth of others. It’s an agar-based medium that is used to isolate microorganisms in labs. The selective growth of microbes is decided by adding substances like antibiotics, dyes, bile salts, or by pH adjustments. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 4. Enrichment media It’s a liquid medium, used to increase the relative concentration of certain microbes before culturing them on a solid medium plate. It’s used as a broth medium and inhibits the growth of commensal species of microorganisms (those who live in close association with each other) in the clinical specimen. It’s also used in isolating fecal and soil microorganisms. Examples are selenite F broth which is used to isolate Salmonella typhi from a fecal sample, tetrathionate broth, and alkaline peptone water. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 5. Differential or indicator media It contains certain indicators like dyes or metabolic substrates in the medium composition which gives different colors to colonies of different microbial species when they utilize or react with these components. It allows the growth of more than one microorganism, however, the bacterial colonies are differentiated based on their color when a chemical change occurs in the indicator, such as neutral red, phenol red, methylene blue. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 6. Transport media: Transport media are useful for clinical specimens which are required to be transferred immediately to labs to maintain the viability of potential pathogens and to prevent overgrowth of commensals or contaminating microorganisms. Some of them are semi-solid in consistency. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 7. Anaerobic media This media is for anaerobic bacteria which require low oxygen levels, extra nutrients, and reduced oxidation- reduction potential. It is supplemented with hemin and vitamin K nutrients and oxygen is removed by boiling it in a water bath and sealing it with paraffin film. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya C. Classification of culture media based on application/ chemical composition 8. Assay media It’s used for amino acids, vitamins, and antibiotics assays. For example, antibiotic assay media is used to determine the antibiotic potency of microorganisms. 9. Storage media It’s used to store microorganisms for a longer period, examples are chalk cooked meat broth and egg saline medium. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Components and Their Roles Peptones and Protein Sources: Derived from proteins, peptones are hydrolysates that provide nitrogen, vitamins, and growth factors essential for microbial growth. Carbohydrates: These serve as energy sources. For example, lactose in MacConkey agar allows for differentiation of lactose-fermenting bacteria. Minerals and Salts: Essential ions like sodium, potassium, magnesium, and phosphate are crucial for various cellular processes and maintaining osmotic balance. Agar: A polysaccharide derived from seaweed, agar is a common solidifying agent. It is not metabolized by most microorganisms, making it ideal for solid media. Buffers: Substances like phosphate or bicarbonate that help maintain a stable pH in the medium. Indicators: pH indicators like phenol red or neutral red change color in response to the pH changes resulting from microbial activity, aiding in differentiation. Selective Agents: These include antibiotics or dyes that inhibit the growth of unwanted microbes. For example, crystal violet in MacConkey agar inhibits Gram-positive bacteria. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Applications in Research and Industry Clinical Diagnostics: Culture media are essential in clinical microbiology for isolating and identifying pathogens from clinical specimens such as blood, urine, or tissue. Pharmaceuticals: Media are used in the production of antibiotics, vitamins, vaccines, and other pharmaceuticals. Fermentation media are critical in this context. Food Microbiology: Used to detect and quantify microbial contaminants in food and beverages. For example, selective media are used to isolate foodborne pathogens like E. coli or Salmonella. Environmental Microbiology: Culture media help in studying microorganisms from soil, water, and air, including the isolation of environmental pathogens or the study of microbial ecology. Biotechnology and Genetic Engineering: Culture media are used to grow genetically modified organisms (GMOs) and for producing enzymes, biofuels, and other industrial products. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Innovations in Culture Media 3D Cell Culture Media: Recently, there has been a move toward 3D cell culture systems that better mimic the natural environment of cells, allowing for more accurate studies in cancer research, drug testing, and tissue engineering. Synthetic and Defined Media: These are formulated with known quantities of all ingredients, making them ideal for controlled studies of microbial physiology or for organisms with specific nutritional requirements. Serum-Free Media: Used in cell culture to avoid the variability and ethical issues associated with animal serum, these media are increasingly used in stem cell research and regenerative medicine. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya Challenges and Considerations Sterility: Ensuring the sterility of culture media is critical to avoid contamination. Autoclaving is the most common method for sterilizing media. Shelf Life: Some media have a limited shelf life, especially those containing labile components like blood or certain vitamins. Cost and Availability: The cost of media can vary significantly, particularly for specialized media or those requiring high-quality ingredients like fetal bovine serum (FBS). Ethical Concerns: The use of animal-derived components in media, such as serum or blood, raises ethical concerns, leading to a push toward more sustainable and humane alternatives. Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya  -----------END--------------- Copyright © 2022 Saint Mary’s University, Bayombong, Nueva Vizcaya