Culture Media by Morphological Classification

Morphological Class

Protozoa

Culture Media

1 Sula’s Medium A liquid medium containing glycerol, asparagine, and a variety of salts. The growth of M. tuberculosis results in turbidity.
2 CHROMagar Candida This differential medium allows for the isolation and identification of Candida species based on colony color. Candida albicans usually forms green colonies on this medium.
3 Potato Dextrose Agar (PDA) Potato Dextrose Agar (PDA) consists of a nutrient-rich substrate, made from dehydrated Potato Infusion and Dextrose, ideal for robust mycological propagation. Agar provides the solidifying medium. Acidification, typically using sterile tartaric acid, adjusts the pH to 3.5 +/- 0.1 to create a more selective environment by inhibiting bacterial proliferation. Additionally, Chloramphenicol is incorporated as an antimicrobial agent to further suppress bacterial contamination, thereby facilitating the selective isolation of fungi.

Mycobacteria

Culture Media

1 Sf1Ep medium Culturing Treponema pallidum, the bacteria that causes syphilis, in vitro is a challenge. T. pallidum is not routinely cultured in the laboratory for diagnostic purposes, in part because it cannot be grown on artificial media. However, a breakthrough in culturing T. pallidum was reported in 2018 when researchers managed to grow the bacterium in a rabbit epithelial cell line (Sf1Ep) using a medium called 'Sf1Ep medium'. It's not used for routine diagnostic purposes, but for research only. The diagnosis involves direct microscopic examination, serologic tests, molecular tests and histopathology.
2 Baird-Parker Agar (BPA) his is a selective medium for the isolation of Staphylococcus species. It has lithium chloride and glycine to inhibit the growth of Gram-negative bacteria and most Gram-positive bacteria except Staphylococcus. Egg yolk emulsion is added to detect lecithinase production and tellurite reduction. S. aureus colonies on BPA are black due to reduction of tellurite, and they exhibit a clear zone due to lecithinase activity on egg yolk.
3 Phenol Red Mannitol Broth This is a differential medium, used to determine an organism's ability to ferment mannitol. S. aureus, which can ferment mannitol, will change the medium from red to yellow due to acid production.
4 DNase Test Agar S. aureus produces the enzyme DNase which hydrolyses DNA. When S. aureus grows on this medium, the DNA is broken down, which can be visualised using a hydrochloric acid (HCl) solution: clear zones around the colonies indicate DNA breakdown.
5 Proskauer and Beck’s Medium A liquid medium. The growth of M. tuberculosis causes turbidity.
6 Dubos’ Medium A liquid medium that contains a mixture of salts, fatty acids, and polysorbate. When M. tuberculosis grows in this medium, it causes the medium to become turbid.
7 Loeffler Medium A serum-based medium. M. tuberculosis colonies on Loeffler medium are small, dry, wrinkled, and off-white to yellow.
8 Petragnini Medium An egg-based medium enriched with additional nutrients to promote the growth of Mycobacterium tuberculosis. Colonies of M. tuberculosis are small, round, buff-colored, and typically take 3-4 weeks to appear.
9 Liquid Media (such as Nutrient Broth) E. coli exhibits homogenous turbid growth within 12-18 hours. After prolonged incubation, pellicles may form on the surface of the media.
10 Urea Agar/Broth E. coli is typically urease negative, so no color change would be expected in this medium.
11 Eosin Methylene Blue (EMB) Agar A selective and differential medium where E. coli forms distinctive metallic green sheen colonies due to vigorous lactose fermentation.
12 CHROMagar Candida This differential medium allows for the isolation and identification of Candida species based on colony color. Candida albicans usually forms green colonies on this medium.
13 Blood Agar A differential medium used to identify bacteria based on their hemolytic properties. E. faecalis typically shows gamma-hemolysis on this medium, i.e., no hemolysis or change in the color of the medium.
14 MacConkey Agar Traditionally used to isolate and differentiate Gram-negative bacilli, E. faecalis can grow on this medium, producing small, round, magenta pink colonies due to lactose fermentation, indicating a positive result.

Fungi

Culture Media

1 Sf1Ep medium Culturing Treponema pallidum, the bacteria that causes syphilis, in vitro is a challenge. T. pallidum is not routinely cultured in the laboratory for diagnostic purposes, in part because it cannot be grown on artificial media. However, a breakthrough in culturing T. pallidum was reported in 2018 when researchers managed to grow the bacterium in a rabbit epithelial cell line (Sf1Ep) using a medium called 'Sf1Ep medium'. It's not used for routine diagnostic purposes, but for research only. The diagnosis involves direct microscopic examination, serologic tests, molecular tests and histopathology.
2 Baird-Parker Agar (BPA) his is a selective medium for the isolation of Staphylococcus species. It has lithium chloride and glycine to inhibit the growth of Gram-negative bacteria and most Gram-positive bacteria except Staphylococcus. Egg yolk emulsion is added to detect lecithinase production and tellurite reduction. S. aureus colonies on BPA are black due to reduction of tellurite, and they exhibit a clear zone due to lecithinase activity on egg yolk.
3 Dorset Medium An egg-based medium. M. tuberculosis colonies appear similar to those on Petragnini Medium: small, round, buff-colored, and taking 3-4 weeks to develop.
4 Middlebrook 7H10 Agar A selective medium that contains oleic acid, albumin, dextrose, and catalase. The colonies of M. tuberculosis appear small, rough, and buff to white-colored, taking less time to appear compared to egg-based media.
5 m-ENDO Agar E. coli colonies appear as green with a metallic sheen, indicating lactose fermentation.
6 Mueller Hinton Agar (MHA) Typically used for antibiotic susceptibility testing. E. coli colonies appear pale straw colored.
7 Nickerson’s Medium or Bismuth Sulfite Glucose Glycine Yeast (BSGG) This medium is used to stimulate the production of germ tubes, a characteristic of Candida albicans.
8 Cornmeal Agar with Tween 80 It is used for the identification of Candida species by promoting the formation of chlamydospores, which is a characteristic of Candida albicans.
9 RPMI 1640 Medium It is a rich medium that contains inorganic salts, glucose, amino acids, vitamins, and other nutrients that promote the growth of yeast cells.
10 Potato Dextrose Agar (PDA) Potato Dextrose Agar (PDA) consists of a nutrient-rich substrate, made from dehydrated Potato Infusion and Dextrose, ideal for robust mycological propagation. Agar provides the solidifying medium. Acidification, typically using sterile tartaric acid, adjusts the pH to 3.5 +/- 0.1 to create a more selective environment by inhibiting bacterial proliferation. Additionally, Chloramphenicol is incorporated as an antimicrobial agent to further suppress bacterial contamination, thereby facilitating the selective isolation of fungi.
11 Sabouraud Dextrose Agar (SDA) It is composed of peptone, dextrose (glucose), and agar. The high dextrose concentration promotes fungal growth, while the acidic pH inhibits bacterial growth.
12 Nutrient Agar It is a general-purpose medium. However, E. faecalis grows poorly on nutrient agar, which means it does not proliferate as well on this medium compared to the others listed, indicating a negative or poor growth.

Bacilli

Culture Media

1 Sf1Ep medium Culturing Treponema pallidum, the bacteria that causes syphilis, in vitro is a challenge. T. pallidum is not routinely cultured in the laboratory for diagnostic purposes, in part because it cannot be grown on artificial media. However, a breakthrough in culturing T. pallidum was reported in 2018 when researchers managed to grow the bacterium in a rabbit epithelial cell line (Sf1Ep) using a medium called 'Sf1Ep medium'. It's not used for routine diagnostic purposes, but for research only. The diagnosis involves direct microscopic examination, serologic tests, molecular tests and histopathology.
2 Baird-Parker Agar (BPA) his is a selective medium for the isolation of Staphylococcus species. It has lithium chloride and glycine to inhibit the growth of Gram-negative bacteria and most Gram-positive bacteria except Staphylococcus. Egg yolk emulsion is added to detect lecithinase production and tellurite reduction. S. aureus colonies on BPA are black due to reduction of tellurite, and they exhibit a clear zone due to lecithinase activity on egg yolk.
3 Phenol Red Mannitol Broth This is a differential medium, used to determine an organism's ability to ferment mannitol. S. aureus, which can ferment mannitol, will change the medium from red to yellow due to acid production.
4 Mannitol Salt Agar (MSA), or Chapman Agar This is a selective and differential medium. The high concentration of NaCl (~7.5%) selects for halophiles, organisms that can tolerate high salt concentrations, thereby favouring the growth of Staphylococcus species. Mannitol is the differential component: S. aureus ferments mannitol, lowering the pH of the medium, which results in a color change of the pH indicator from red to yellow. Thus, growth of S. aureus is indicated by yellow colonies.
5 Sauton’s Medium A liquid medium that lacks detergents, which helps in the formation of corded colonies. When M. tuberculosis grows in this medium, it results in turbidity.
6 Middlebrook 7H9 Broth This is a liquid medium that contains glycerol and Tween 80, which prevent clumping of mycobacteria. The growth of M. tuberculosis results in turbidity.
7 Dubos’ Medium A liquid medium that contains a mixture of salts, fatty acids, and polysorbate. When M. tuberculosis grows in this medium, it causes the medium to become turbid.
8 Dorset Medium An egg-based medium. M. tuberculosis colonies appear similar to those on Petragnini Medium: small, round, buff-colored, and taking 3-4 weeks to develop.
9 Middlebrook 7H10 Agar A selective medium that contains oleic acid, albumin, dextrose, and catalase. The colonies of M. tuberculosis appear small, rough, and buff to white-colored, taking less time to appear compared to egg-based media.
10 Lowenstein-Jensen (LJ) Medium An egg-based medium that uses malachite green to suppress the growth of other bacteria and glycerol to stimulate the growth of Mycobacterium tuberculosis. The colonies of M. tuberculosis on LJ are non-pigmented, dry, rough, raised, irregular with a wrinkled surface, initially creamy-white, turning yellowish or buff-colored on further incubation.
11 Blood Agar (BA) E. coli colonies on this differential medium are large, circular, gray, moist, and can show β-hemolysis.
12 Cornmeal Agar with Tween 80 It is used for the identification of Candida species by promoting the formation of chlamydospores, which is a characteristic of Candida albicans.
13 RPMI 1640 Medium It is a rich medium that contains inorganic salts, glucose, amino acids, vitamins, and other nutrients that promote the growth of yeast cells.
14 Potato Dextrose Agar (PDA) Potato Dextrose Agar (PDA) consists of a nutrient-rich substrate, made from dehydrated Potato Infusion and Dextrose, ideal for robust mycological propagation. Agar provides the solidifying medium. Acidification, typically using sterile tartaric acid, adjusts the pH to 3.5 +/- 0.1 to create a more selective environment by inhibiting bacterial proliferation. Additionally, Chloramphenicol is incorporated as an antimicrobial agent to further suppress bacterial contamination, thereby facilitating the selective isolation of fungi.

Cocci

Culture Media

1 CHROMagar Candida This differential medium allows for the isolation and identification of Candida species based on colony color. Candida albicans usually forms green colonies on this medium.
2 Potato Dextrose Agar (PDA) Potato Dextrose Agar (PDA) consists of a nutrient-rich substrate, made from dehydrated Potato Infusion and Dextrose, ideal for robust mycological propagation. Agar provides the solidifying medium. Acidification, typically using sterile tartaric acid, adjusts the pH to 3.5 +/- 0.1 to create a more selective environment by inhibiting bacterial proliferation. Additionally, Chloramphenicol is incorporated as an antimicrobial agent to further suppress bacterial contamination, thereby facilitating the selective isolation of fungi.

Spirillum

Culture Media

1 Nickerson’s Medium or Bismuth Sulfite Glucose Glycine Yeast (BSGG) This medium is used to stimulate the production of germ tubes, a characteristic of Candida albicans.
2 RPMI 1640 Medium It is a rich medium that contains inorganic salts, glucose, amino acids, vitamins, and other nutrients that promote the growth of yeast cells.
3 Potato Dextrose Agar (PDA) Potato Dextrose Agar (PDA) consists of a nutrient-rich substrate, made from dehydrated Potato Infusion and Dextrose, ideal for robust mycological propagation. Agar provides the solidifying medium. Acidification, typically using sterile tartaric acid, adjusts the pH to 3.5 +/- 0.1 to create a more selective environment by inhibiting bacterial proliferation. Additionally, Chloramphenicol is incorporated as an antimicrobial agent to further suppress bacterial contamination, thereby facilitating the selective isolation of fungi.
4 Azide Dextrose Broth A selective medium inhibiting Gram-negative bacteria, used for the isolation of streptococci and staphylococci from mixed samples. E. faecalis will show a positive growth resulting in a turbid appearance of the broth.
5 Bile Esculin Agar (BEA) A selective and differential medium that differentiates group D Streptococci and Enterococci based on the ability to hydrolyze esculin in the presence of bile. E. faecalis hydrolyzes esculin, leading to the formation of a dark brown or black precipita

Spirochetes

Culture Media

1 Phenol Red Mannitol Broth This is a differential medium, used to determine an organism's ability to ferment mannitol. S. aureus, which can ferment mannitol, will change the medium from red to yellow due to acid production.
2 DNase Test Agar S. aureus produces the enzyme DNase which hydrolyses DNA. When S. aureus grows on this medium, the DNA is broken down, which can be visualised using a hydrochloric acid (HCl) solution: clear zones around the colonies indicate DNA breakdown.
3 BACTEC This is not a traditional medium but a system for detecting the growth of M. tuberculosis by monitoring the release of C14O2 from C14 palmitic acid, which the bacteria metabolize. An increase in radioactive counts in the BACTEC instrument indicates the growth of bacteria.
4 Pawlowsky Medium A potato-based medium. Growth of M. tuberculosis may be similar to that seen on the LJ Medium.
5 Brilliant Green Agar (BGA) A selective medium that is used to isolate Salmonella species, but E. coli can grow on it, albeit not as well.
6 Urea Agar/Broth E. coli is typically urease negative, so no color change would be expected in this medium.
7 Nutrient Agar It is a general-purpose medium. However, E. faecalis grows poorly on nutrient agar, which means it does not proliferate as well on this medium compared to the others listed, indicating a negative or poor growth.
8 Bile Esculin Agar (BEA) A selective and differential medium that differentiates group D Streptococci and Enterococci based on the ability to hydrolyze esculin in the presence of bile. E. faecalis hydrolyzes esculin, leading to the formation of a dark brown or black precipita