Microbial Culture Media- Definition, Types, Examples, Uses
Culture media are substances that provide the necessary nutrients and environmental conditions for the growth and maintenance of microorganisms in vitro. Microorganisms have diverse nutritional and physiological requirements, and different types of culture media are designed to meet these needs. Culture media can be classified based on various criteria, such as their composition, consistency, chemical properties, application, and oxygen requirement. In this article, we will discuss the definition, types, examples, and uses of different cultural media used in microbiology.
The main purpose of culture media is to support the growth of microorganisms and to allow their observation and identification. Culture media can also be used to study the biochemical, metabolic, and physiological characteristics of microorganisms, as well as their interactions with other organisms or substances. Culture media can also be used to test the antimicrobial susceptibility, virulence, or genetic traits of microorganisms.
The basic components of culture media are water, carbon source, nitrogen source, minerals, vitamins, and growth factors. Depending on the type of microorganism and the objective of the culture, additional ingredients may be added to the media, such as agar, blood, serum, egg yolk, antibiotics, dyes, indicators, or inhibitors. These ingredients can modify the physical state, chemical composition, or selective and differential properties of the media.
The preparation of culture media requires careful attention to the quality and quantity of the ingredients, the sterilization method, the pH adjustment, and the storage conditions. Culture media should be prepared according to standard protocols and quality control measures to ensure their reliability and reproducibility. Cultural media should also be labeled with the name, date of preparation, expiration date, and any other relevant information.
Cultural media play a vital role in microbiology research and diagnostics. They enable the isolation, cultivation, identification, and characterization of microorganisms from various sources. They also facilitate the investigation of microbial diversity, ecology, evolution, and pathogenesis. Culture media are essential tools for microbiologists to explore the fascinating world of microorganisms.
Culture media are substances that provide nutrients and environmental conditions for the growth of microorganisms in vitro. Different types of culture media are used for different purposes, such as isolation, identification, cultivation, or biochemical testing of microorganisms. Culture media can be classified based on their composition, consistency, chemical properties, or application.
One of the main factors that determine the composition of a culture medium is the nutritional requirements of the microorganisms to be cultured. Some microorganisms are fastidious, meaning they need specific and complex nutrients to grow, while others are non-fastidious, meaning they can grow on simple and general nutrients. Therefore, culture media may vary in their complexity and specificity depending on the target microorganisms.
However, there are some common ingredients that are found in most cultural media, regardless of their type or purpose. These ingredients provide the basic constituents for microbial growth, such as water, carbon, nitrogen, energy, minerals, vitamins, and buffering agents. Here are some examples of common ingredients of cultural media and their functions:
- Water: It is an essential ingredient of any culture medium, as it is the source of hydrogen and oxygen for microbial metabolism. Water also dissolves and transports other nutrients and wastes in the medium. Deionized or distilled water is usually used to prepare culture media to avoid contamination or interference from impurities.
- Agar: It is a polysaccharide extracted from red algae that act as a solidifying agent for culture media. Agar has several advantages over other gelling agents, such as gelatin or starch. Agar is inert, meaning it does not react with or inhibit microbial growth. Agar also has a high melting point (about 100°C) and a low solidifying point (about 45°C), which allows easy sterilization and manipulation of culture media. Agar is usually added at a concentration of 1.5-2% to solidify liquid media.
- Peptone: It is a complex mixture of partially digested proteins that provides a source of carbon and nitrogen for microbial growth. Peptone is derived from animal or plant sources, such as meat, casein, soybean, or wheat. Peptone contains amino acids, peptides, and other organic compounds that can be utilized by various microorganisms. Peptone is usually added at a concentration of 0.5-1% to culture media.
- Meat extract: It is a concentrated solution of soluble substances extracted from animal tissues, such as beef or pork. Meat extract provides a source of amino acids, vitamins, minerals, and other growth factors for microorganisms. Meat extract also enhances the flavor and color of culture media. Meat extract is usually added at a concentration of 0.1-0.5% to culture media.
- Yeast extract: It is a similar product to meat extract but derived from yeast cells instead of animal tissues. Yeast extract provides a source of vitamin B complex, nucleic acids, carbohydrates, and other growth factors for microorganisms. Yeast extract also stimulates the growth of bacteria by providing them with carbon and nitrogen sources. Yeast extract is usually added at a concentration of 0.1-0.5% to culture media.
- Mineral salts: These are inorganic compounds that provide essential elements for microbial metabolisms, such as magnesium, potassium, iron, calcium, phosphorus, sulfur, etc. Mineral salts also help maintain the osmotic balance and pH of the medium. Mineral salts are usually added at low concentrations (less than 0.1%) to culture media.
- Carbohydrates: They are organic compounds that provide a source of energy and carbon for microorganisms. Carbohydrates can also serve as indicators or substrates for biochemical reactions or fermentation processes. Common carbohydrates used in culture media include glucose, lactose, sucrose, maltose, etc. Carbohydrates are usually added at a concentration of 0.5-1% to culture media.
- Buffering system: It is a combination of substances that resist changes in pH when acids or bases are added to the medium. The buffering system is important for maintaining the optimal pH range for microbial growth and activity. Common buffering agents used in culture media include phosphate salts, acetate salts, citrate salts, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), etc.
- pH indicator: It is a substance that changes color depending on the pH of the medium. pH indicator is useful for detecting acid or alkaline production by microorganisms or for monitoring the pH changes during microbial growth or biochemical reactions. Common pH indicators used in culture media include phenol red, bromothymol blue, neutral red, etc.
These are some examples of common ingredients of culture media that provide the basic components for microbial growth and differentiation. However, depending on the type and purpose of the culture medium, other ingredients may be added or modified to enhance or inhibit certain microorganisms or characteristics. For example:
- Serum: It is a liquid fraction of blood that contains proteins (such as albumin), antibodies (such as immunoglobulins), hormones (such as insulin), electrolytes (such as sodium), etc. Serum enriches the medium for fastidious or pathogenic microorganisms that require specific nutrients or factors for growth. For example, blood agar contains 5% sheep blood; chocolate agar contains heated blood; LSS agar contains serum.
- Antibiotics: They are substances that inhibit or kill bacteria by interfering with their cell wall synthesis (such as penicillin), protein synthesis (such as tetracycline), DNA replication (such as quinolone), etc. Antibiotics make the medium selective for certain microorganisms by suppressing the growth of others. For example, MacConkey agar contains bile salts and crystal violet; Thayer-Martin agar contains vancomycin,
Columbia-colistin-nalidixic acid agar contains colistin
and nalidixic acid.
- Dyes: They are substances that stain or color microorganisms or their products based on their chemical properties or interactions. Dyes make the medium differential for certain microorganisms by showing visible changes in their appearance. For example, Eosin-methylene blue agar contains eosin
and methylene blue;
mannitol salt agar contains phenol red;
MacConkey agar contains neutral red.
These are some examples of additional ingredients that can modify the common ingredients of culture media to suit different purposes or applications. Therefore,
culture media can be customized according to the nutritional,
and biochemical characteristics
of the target microorganisms.
Culture media are fundamental in diagnostic microbiology. They provide nutrients and environmental conditions to support the growth of microorganisms in vitro. Culture media can be solid (agar-based) or liquid (broth-based), depending on the purpose and method of inoculation. Some culture media are also selective, differential, or enriched to favor or distinguish certain microorganisms from others.
Here is a general procedure for preparing cultural media :
- Obtain the necessary ingredients and equipment, such as a sterile container, agar or broth, and any supplements or growth factors that the microorganism requires.
- Measure out the appropriate amount of ingredients and mix them together in a sterile container. Follow the manufacturer`s instructions for the amount and type of water to use, as well as the pH adjustment if needed. For agar media, add agar powder and boil it to dissolve completely.
- Sterilize the media by autoclaving at 121°C for 15 minutes at 15 psi. This will kill any contaminating microorganisms and ensure the media are safe to use. Alternatively, some media can be sterilized by filtration using a 0.22-micron filter.
- Cool down the media to around 50°C for agar media or room temperature for broth media. For agar media, pour them into sterile Petri plates or tubes and allow them to solidify. For broth media, dispense them into sterile tubes or flasks.
- Label the media with the name, date, lot number, and expiry date. Store them in appropriate conditions according to the manufacturer`s recommendations. For example, some media may need to be refrigerated or protected from light.
- Perform quality control tests on the media before use. This may include checking for sterility, pH, appearance, color, clarity, and performance with known microorganisms.
Some tips for preparing cultural media are:
- Use high-quality dehydrated media powder from reputable manufacturers and store them in humidity-proof packaging.
- Use distilled or deionized water to dissolve the media powder and avoid tap water that may contain chlorine or minerals that can affect the media quality.
- Use clean and dry glassware and equipment, and avoid metal utensils that can react with some media components.
- Avoid overheating or overcooking the media, as this can damage some nutrients or indicators.
- Avoid repeated heating and cooling cycles of agar media as this can cause condensation or evaporation that can alter the concentration of the media.
- Avoid contamination of the media by using aseptic techniques and working in a laminar flow hood or biosafety cabinet if possible.
Types of culture media based on composition
Cultural media can be classified based on their chemical composition:
- Defined media: These are chemically defined media that contain known amounts and types of pure organic and inorganic compounds. These media are used to cultivate fastidious microorganisms that require specific nutrients for growth. For example, Czapek Dox medium is a defined medium that contains glucose, sodium nitrate, magnesium sulfate, potassium chloride, ferrous sulfate, and dipotassium phosphate.
- Undefined media: These are complex media that contain various ingredients of unknown composition and quantity, such as yeast extract, beef extract, peptone, etc. These media are used to grow non-fastidious microorganisms that can utilize a variety of nutrients. For example, nutrient agar, MacConkey agar, and blood agar are undefined media that contain beef extract, peptone, agar, and other additives.
- Complex media: These are media that are modified from basal or undefined media by adding additional substances to enhance the growth of specific microorganisms or to bring out certain characteristics of the microorganisms. For example, chocolate agar is a complex medium that is prepared by heating blood agar to release hemin and NAD factors that are essential for the growth of Haemophilus and Neisseria species.
Culture media can be classified based on their consistency or physical state into three types: solid, semi-solid, and liquid media. The consistency of the media depends on the amount of agar or other solidifying agent added to the media. Agar is a polysaccharide extracted from red algae that can form a gel-like matrix when dissolved in water and cooled. Agar is commonly used as a solidifying agent because it is not degraded by most microorganisms, and it remains solid at temperatures suitable for microbial growth.
Solid media are prepared by adding agar at a concentration of 1.5-2.0% to the liquid broth. Solid media solidify at around 37°C and can be poured into Petri dishes or tubes to form slants or stabs. Solid media are used for isolating bacteria as pure cultures, studying colony morphology, performing antimicrobial susceptibility testing, and differentiating bacteria based on their biochemical reactions. Examples of solid media are nutrient agar, MacConkey agar, blood agar, chocolate agar, and mannitol salt agar. The growth of bacteria on solid media can vary in appearance, such as smooth, rough, mucoid, round, irregular, filamentous, or punctiform
Semi-solid media are prepared by adding agar at a concentration of 0.5% or less to the liquid broth. Semi-solid media have a soft custard-like consistency and do not hold their shape well. Semi-solid media are used for demonstrating bacterial motility and cultivating microaerophilic bacteria. Examples of semi-solid media are Stuart
s and Amies transport media, Hugh and Leifsons oxidation fermentation medium, and mannitol motility medium. The growth of bacteria in semi-solid media appears as a thick line or a diffuse cloud in the medium.
Liquid media are prepared without any solidifying agent and have a clear or turbid appearance depending on the ingredients. Liquid media are also called broths and are used for growing large numbers of bacteria, studying fermentation reactions, performing serological tests, and cultivating anaerobic bacteria. Examples of liquid media are nutrient broth, tryptic soy broth, MR-VP broth, phenol red carbohydrate broth, and thioglycolate broth. The growth of bacteria in liquid media can be detected by turbidity, sedimentation, pellicle formation, or color change in the medium.
There are seven routine laboratory media based on their chemical composition and application. They are:
- Basal media
- Enriched media
- Selective media
- Enrichment media
- Indicator or differential media
- Transport media
- Storage media
Basal media are simple media that provide the basic nutrients for the growth of many microorganisms. They contain carbon and nitrogen sources, such as peptone, beef extract, yeast extract, and various salts. They are not selective or differential and allow the growth of non-fastidious bacteria without any enrichment source. They are used for sub-culturing and routine laboratory work. Examples of basal media are Nutrient Agar, Peptone Water, etc.
Enriched media are basal media that have been supplemented with additional substances, such as blood, serum, egg yolk, or other growth factors. These media enhance the growth of fastidious microorganisms that require specific nutrients or conditions for growth. They may also inhibit the growth of unwanted microorganisms by creating a selective environment. Examples of enriched media are Blood Agar, Chocolate Agar, Lowenstein Jensen Media, etc.
Selective media are media that allow the growth of certain microorganisms while inhibiting the growth of others. They achieve this by adding selective agents, such as bile salts, antibiotics, dyes, pH adjustments, etc., to the medium. These agents target specific characteristics of the desired or undesired microorganisms, such as cell wall structure, metabolic pathways, enzyme production, etc. Selective media are useful for isolating microorganisms from mixed specimens or sources. Examples of selective media are MacConkey Agar, Eosin Methylene Blue Agar, Mannitol Salt Agar, etc.
Enrichment media are liquid media that enhance the growth of desired microorganisms at a low density. They provide an environment and conditions similar to selective media and inhibit unwanted bacteria from growing. They are used for isolating microorganisms from soil, feces, water, or other sources where they may be present in small numbers. Examples of enrichment media are Selenite F Broth, Alkaline Peptone Water, Tetrathionate Broth, etc.
Indicator or differential media
Indicators or differential media are media that show visible changes due to the presence of an indicator. They differentiate bacteria based on colony color, shape, size, or other characteristics that reflect their biochemical or physiological properties. They may also contain selective agents to inhibit unwanted microorganisms. Examples of indicator or differential media are Mannitol Salt Agar (mannitol fermentation shows yellow color colonies), Blood Agar (hemolysis shows clear or green zones around colonies), MacConkey Agar (lactose fermentation shows pink colonies), etc.
Transport media are media that preserve and maintain the viability of microorganisms during transport from the specimen collection site to the laboratory. They prevent the microorganisms from drying out, dying, or multiplying during transit. They may also contain buffers, antibiotics, or other agents to reduce pH changes or contamination. Examples of transport media are Stuart
s Transport Medium (lacks carbon and nitrogen sources), Cary Blairs Transport Medium (contains sodium thioglycolate and sodium glycerophosphate), Amies Transport Medium (contains charcoal), etc.[^1] 
Storage media are media that prolong the longevity of bacterial cultures[^1]^ ^. They contain nutrients and substances that protect the bacteria from adverse conditions such as temperature changes, desiccation, oxidation, etc.^ . They are used for preserving bacterial strains for future use or reference[^1]^. Examples of storage media are Cooked Meat Broth (contains meat particles and glucose), Nutrient Agar with 20% Glycerol (contains glycerol as a cryoprotectant), Skim Milk Broth (contains casein as a protein source), etc.^ .
Types of culture media based on oxygen requirement
Microorganisms have different requirements for growth depending on their oxygen needs. Some microbes can use oxygen as a terminal electron acceptor in aerobic respiration, while others can use alternative electron acceptors such as nitrate, sulfate, or carbon dioxide in anaerobic respiration. Some microbes can switch between aerobic and anaerobic respiration depending on the availability of oxygen, while others cannot tolerate oxygen at all and use fermentation to produce energy. Therefore, culture media must be designed to provide the appropriate oxygen conditions for the growth of different microbes.
Aerobic media are culture media that allow the diffusion of oxygen throughout the medium. They are suitable for the cultivation of obligate aerobes, which require oxygen for growth, and facultative anaerobes, which can grow with or without oxygen. Aerobic media can be solid or liquid and usually contain peptone, beef extract, yeast extract, and agar as basic ingredients. Examples of aerobic media are:
- Nutrient agar: a general-purpose medium used for the isolation of a wide variety of bacteria.
- Blood agar: a differential medium that contains 5% sheep blood and can show hemolytic reactions of bacteria.
- Chocolate agar: a medium that contains lysed blood cells and is enriched for fastidious bacteria such as Neisseria and Haemophilus.
- MacConkey agar: a selective and differential medium that contains bile salts and crystal violet to inhibit gram-positive bacteria and lactose and neutral red to differentiate lactose fermenters from non-fermenters among gram-negative bacteria.
Anaerobic media are culture media that prevent the diffusion of oxygen throughout the medium. They are suitable for the cultivation of obligate anaerobes, which cannot grow in the presence of oxygen, and aerotolerant anaerobes, which can survive but not use oxygen. Anaerobic media can be solid or liquid and usually contain peptone, beef extract, yeast extract, cysteine, sodium thioglycolate, hemin, vitamin K, and agar as basic ingredients. Examples of anaerobic media are:
- Thioglycolate broth: a liquid medium that contains sodium thioglycolate as a reducing agent to bind any free oxygen in the medium. It also has a low concentration of agar to prevent convection currents that could bring oxygen into the medium. The medium is boiled before use to expel any dissolved oxygen and sealed with a screw cap to prevent oxygen entry. An indicator such as methylene blue or resazurin is added to show the presence or absence of oxygen by changing color. The growth of bacteria in thioglycollate broth can indicate their oxygen requirements: obligate aerobes grow near the surface where oxygen is available, obligate anaerobes grow near the bottom where oxygen is absent, facultative anaerobes grow throughout the medium but more densely near the surface, and microaerophiles grow just below the surface where oxygen is reduced.
- Brucella agar: a solid medium that contains hemin and vitamin K as growth factors for anaerobes. It also has sodium bicarbonate to buffer the pH and neutralize any acidic products from fermentation. The medium is boiled before use to expel any dissolved oxygen and poured into sterile Petri plates under anaerobic conditions. The plates are then incubated in an anaerobic jar or chamber that contains a gas mixture of nitrogen, carbon dioxide, and hydrogen (80:10:10) and a catalyst such as palladium to remove any residual oxygen by combining it with hydrogen to form water.
- Bacteroides bile esculin agar: a selective and differential medium that contains bile salts to inhibit most gram-positive bacteria and esculin from differentiating Bacteroides species from other anaerobes. Bacteroides species can hydrolyze esculin to form esculetin, which reacts with ferric citrate in the medium to produce a black precipitate.
Microaerophilic media are culture media that allow a reduced level of oxygen (5-10%) throughout the medium. They are suitable for the cultivation of microaerophiles, which require low concentrations of oxygen for growth. Microaerophilic media can be solid or liquid and usually contain peptone, beef extract, yeast extract, agar, and sodium thioglycolate as basic ingredients. Examples of microaerophilic media are:
- Campylobacter blood agar: a solid medium that contains 5% sheep blood and antibiotics such as vancomycin, polymyxin B, trimethoprim, and amphotericin B to inhibit most bacteria except Campylobacter species. The medium is boiled before use to expel any dissolved oxygen and poured into sterile Petri plates under microaerophilic conditions. The plates are then incubated in a microaerophilic jar or chamber that contains a gas mixture of nitrogen, carbon dioxide, hydrogen, and oxygen (85:10:5:5) and a catalyst such as palladium to adjust the oxygen level by combining it with hydrogen to form water.
- Skirrow medium: a solid medium that contains 5% horse blood and antibiotics such as vancomycin, polymyxin B, and trimethoprim-sulfamethoxazole to inhibit most bacteria except Campylobacter species. The medium is boiled before use to expel any dissolved oxygen and poured into sterile Petri plates under microaerophilic conditions. The plates are then incubated in a microaerophilic jar or chamber that contains a gas mixture of nitrogen, carbon dioxide, hydrogen, and oxygen (85:10:5:5) and a catalyst such as palladium to adjust the oxygen level by combining it with hydrogen to form water.
Capnophilic media are culture media that allow an elevated level of carbon dioxide (5-10%) throughout the medium. They are suitable for the cultivation of capnophiles, which require high concentrations of carbon dioxide for growth. Capnophilic media can be solid or liquid and usually contain peptone, beef extract, yeast extract,
agar, sodium bicarbonate, or sodium carbonate as basic ingredients. Examples of capnophilic media are:
- Columbia agar with 5% sheep blood: a general-purpose medium used for the isolation of a variety of bacteria,
including fastidious organisms such as Streptococcus pneumoniae and Neisseria meningitides that are capnophilic.
The medium is boiled before use to expel any dissolved oxygen and poured into sterile Petri plates under
capnophilic conditions. The plates are then incubated in a capnophilic jar or chamber that contains a gas mixture
of nitrogen and carbon dioxide (90:10) or air supplemented with carbon dioxide (5-10%).
- Chocolate agar: a medium that contains lysed blood cells and is enriched for fastidious bacteria, such as Neisseria
and Haemophilus, that are capnophilic. The medium is boiled before use to expel any dissolved oxygen and poured
into sterile Petri plates under capnophilic conditions. The plates are then incubated in a capnophilic jar or chamber
that contains a gas mixture of nitrogen and carbon dioxide (90:10) or air supplemented with carbon dioxide (5-
Special purpose culture media: Assay, Minimal, and Fermentation media
Special-purpose culture media are designed for specific applications or purposes that cannot be achieved by general or selective media. Some examples of special-purpose media are assay media, minimal media, and fermentation media.
Assay media are used to measure the activity or potency of certain substances, such as vitamins, amino acids, antibiotics, and hormones. These substances are usually added to a defined medium that lacks the substance to be assayed, and the growth or metabolic response of the microorganisms is observed. For example, the antibiotic sensitivity test uses Mueller-Hinton agar as an assay medium to determine the susceptibility of bacteria to different antibiotics by measuring the zone of inhibition around antibiotic disks. Another example is the use of Lactobacillus MRS broth as an assay medium to measure the vitamin B12 content of food samples by inoculating them with Lactobacillus leishmania and measuring the turbidity after incubation.
Minimal media are defined as media that contain the minimal amount of nutrients required for the growth of a specific microorganism. They usually consist of a carbon source, such as glucose or succinate, and inorganic salts, such as magnesium, nitrogen, sulfur, and phosphorus. Minimal media are used for various purposes, such as studying the nutritional requirements of microorganisms, selecting recombinant bacteria that carry plasmids with certain genes, and testing the ability of microorganisms to utilize different carbon sources. For example, the Czapek Dox medium is a minimal medium that contains sucrose as the sole carbon source and sodium nitrate as the sole nitrogen source. It is used for the cultivation and differentiation of fungi based on their ability to produce pigments.
Fermentation media are media that support the growth and metabolic activity of microorganisms that produce desirable products by fermentation. Fermentation is a process in which microorganisms convert organic substrates into simpler compounds, such as ethanol, lactic acid, acetic acid, or hydrogen gas, under anaerobic or low-oxygen conditions. Fermentation media usually contain high levels of nutrients, such as carbon, nitrogen, vitamins, minerals, and growth factors, as well as specific substrates for the synthesis of products in the fermentation process. For example, yeast extract is a common ingredient in fermentation media because it provides various nutrients and stimulates enzyme production in yeast cells. Another example is wort, which is a liquid medium derived from malted barley that is used for the production of beer by yeast fermentation.
Culture media are essential tools for microbiology, as they provide the necessary nutrients and conditions for the growth and isolation of microorganisms from various sources. Culture media can have different applications depending on their composition, consistency, chemical properties, oxygen requirements, and purpose. Some of the common applications of cultural media are:
- To culture microbes for identification, diagnosis, or research purposes.
- To test the antimicrobial susceptibility or resistance of microbes to different agents.
- To observe the biochemical reactions, metabolic products, or physiological characteristics of microbes.
- To differentiate between different types of microbes based on their colony morphology, color, hemolysis, or other indicators.
- To select specific types of microbes by inhibiting or suppressing the growth of others.
- To enrich rare or fastidious microbes by providing additional nutrients or growth factors.
- To transport and store microbes without losing their viability or activity.
However, culture media also have some limitations that need to be considered when using them for microbiological purposes. Some of the common limitations of cultural media are:
- Risk of contamination by unwanted microbes or substances that can affect the results or quality of the culture.
- High skill and expertise required for preparing, sterilizing, inoculating, incubating, and interpreting the culture media.
- Increased cost and time involved in obtaining, maintaining, and disposing of the culture media and related materials.
- Possibility of drying out, deterioration, or alteration of the culture media due to improper storage or handling conditions.
- Variability in the composition, quality, or performance of the culture media due to different sources, batches, or formulations.
- Inability to mimic the natural environment or conditions of some microbes that may affect their growth or behavior in vitro.
- Inadequacy of some culture media to support the growth or detection of some microbes that have complex or unknown nutritional requirements.
Culture media are substances that provide nutrients and conditions for the growth and identification of microorganisms. There are different types of culture media based on their composition, consistency, chemical properties, oxygen requirements, and applications. Here, we will list some of the common culture media used in microbiology with their uses.
Solid or Agar media
Solid media contain agar, which is a polysaccharide extracted from red algae that forms a gel-like structure when dissolved in water and cooled. Agar provides a solid surface for the microorganisms to grow as discrete colonies that can be easily observed and isolated. Solid media can be further classified into different types based on their composition and purpose.
- Nutrient agar: A simple and general-purpose medium that contains peptone, beef extract, sodium chloride, and agar. It supports the growth of many non-fastidious bacteria and is used for routine cultivation and maintenance of bacterial cultures.
- Blood agar: An enriched medium that contains 5% sheep blood in addition to the nutrient agar base. It is used for the isolation and differentiation of fastidious bacteria, such as streptococci and Haemophilus, based on their hemolytic patterns.
- Chocolate agar: A variant of blood agar that contains lysed red blood cells, which release hemin (factor X) and NAD (factor V) that are required by some bacteria, such as Neisseria and Haemophilus. It is used for the isolation and detection of these fastidious organisms.
- MacConkey agar: A selective and differential medium that contains bile salts and crystal violet, which inhibit the growth of gram-positive bacteria, and lactose and neutral red, which differentiate lactose-fermenting bacteria (pink colonies) from non-lactose fermenters (colorless colonies). It is used for the isolation and differentiation of enteric pathogens, such as Escherichia coli and Salmonella.
- Mannitol salt agar: A selective and differential medium that contains 7.5% sodium chloride, which selects for salt-tolerant bacteria, such as staphylococci, and mannitol and phenol red, which differentiate mannitol-fermenting bacteria (yellow colonies) from non-mannitol fermenters (red colonies). It is used for the isolation and identification of Staphylococcus aureus.
- Eosin methylene blue (EMB) agar: A selective and differential medium that contains eosin and methylene blue dyes, which inhibit the growth of gram-positive bacteria and differentiate lactose-fermenting bacteria (dark purple or green colonies with a metallic sheen) from non-lactose fermenters (colorless or pink colonies). It is used for the isolation and differentiation of enteric pathogens, especially E. coli.
- Hektoen enteric (HE) agar: A selective and differential medium that contains bile salts, which inhibit the growth of most gram-positive bacteria and some gram-negative bacteria, and lactose, sucrose, salicin, ferric ammonium citrate, sodium thiosulfate, bromthymol blue, and acid fuchsin, which differentiate lactose/sucrose/salicin-fermenting bacteria (salmon or orange colonies) from non-fermenters (blue-green colonies with or without black centers). It is used for the isolation and differentiation of Salmonella and Shigella from stool specimens.
- Xylose lysine deoxycholate (XLD) agar: A selective and differential medium that contains sodium deoxycholate, which inhibits the growth of gram-positive bacteria and some gram-negative bacteria, and xylose, lysine, lactose, sucrose, sodium thiosulfate, ferric ammonium citrate, phenol red, which differentiate xylose-fermenting bacteria (yellow colonies) from non-xylose fermenters (red colonies), lysine-decarboxylating bacteria (red colonies with purple centers) from non-decarboxylation (red colonies), hydrogen sulfide-producing bacteria (black centers) from non-producers (no blackening). It is used for the isolation and differentiation of enteric pathogens, especially Shigella.
- Cetrimide agar: A selective medium that contains cetrimide, which inhibits the growth of most bacteria except for Pseudomonas aeruginosa. It also contains glycerol as a carbon source and magnesium chloride and potassium sulfate as cofactors for the production of pyocyanin pigment by P. aeruginosa. It is used for the identification of P. aeruginosa.
- Lowenstein-Jensen (LJ) medium: An enriched medium that contains egg albumin, malachite green dye, glycerol or pyruvate as a carbon source, asparagine as a nitrogen source, potassium phosphate buffer, magnesium sulfate as a cofactor for enzymes involved in mycobacterial metabolism. It is used for the isolation and cultivation of Mycobacterium species.
- Sabouraud dextrose agar: A medium that contains peptone, dextrose, and agar at a low pH (5.6), which inhibits the growth of most bacteria but favors the growth of fungi. It is used for the isolation and cultivation of yeasts and molds.
Liquid media are nutrient-rich liquids that are used to grow microorganisms in suspension. They do not contain agar or any other solidifying agent. Liquid media can also be classified into different types based on their composition and purpose.
- Nutrient broth: A simple liquid medium that contains peptone,
and sodium chloride. It supports the growth of many non-fastidious bacteria
and is used for routine cultivation
of bacterial cultures.
- Tryptic soy broth: A general-purpose liquid medium that contains tryptone,
and sodium chloride. It supports the growth
of a wide variety
and anaerobic bacteria,
and molds. It is also recommended by the CLSI
for the preparation
for Kirby-Bauer disk diffusion susceptibility testing
and is the CLSI`s choice
as a sterility testing medium.
- Brain heart infusion broth: A rich liquid medium that contains brain
and heart infusions,
and sodium chloride. It supports the growth
of fastidious organisms,
such as streptococci,
and brucellae. Formulations with 0.1% agar
that reduce O2 tension favor anaerobes,
and formulations with Fildes enrichment are used
for the isolation
of fastidious organisms such
- Thioglycolate broth: An enriched liquid medium that contains casein digest,
and L-cystine. It supports the growth
and anaerobic organisms by reducing oxygen tension
and oxidation-reduction potential.
It also contains hemin
and vitamin K
that are required by some bacteria. The resazurin dye acts
as an indicator
of oxygen presence by changing color from colorless to pink when oxidized.
- Selenite broth: An enrichment broth medium that contains peptone,
and sodium phosphate buffer.
It inhibits the growth
of most intestinal flora except for Salmonella
and Shigella species.
It is used for the isolation
of these enteric pathogens from fecal specimens.
- Alkaline peptone water: An enrichment broth medium that contains peptone,
and sodium hydroxide.
It enhances the growth
of Vibrio cholerae by providing alkaline pH
and high osmotic pressure.
It is used for the isolation
of V. cholerae from stool specimens.
Special purpose media
Special-purpose media are designed to serve specific functions or applications in microbiology. They may have unique ingredients or formulations that enable certain biochemical reactions or physiological processes to occur. They may also have special indicators or additives that facilitate the identification or differentiation of microorganisms.
Some examples of special-purpose media are:
- Assay media: Media that are used to measure the activity or potency of antibiotics,
or other substances by observing their effects on microbial growth.
antibiotic sensitivity test uses Mueller-Hinton agar with different concentrations
of antibiotics to determine their minimum inhibitory concentrations (MICs)
for various bacteria.
- Minimal media: Media that contain only the essential nutrients required by a microorganism to grow.
They have usually defined media with known quantities
of all ingredients.
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