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Bacterial Culture Media: Types and Applications Flipbook PDF
Name: Muhammad Khuzaifah bin Mohd Atan Student's ID: 2019488194 Group: HS2412A
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Bacterial Culture Media TYPES and APPLICATIONS MICROBIOLOGY I | MLT 422 Muhammad Khuzaifah bin Mohd Atan 2019488194 HS2412A Dr Siti Nazrina Camalxaman
TABLE OF CONTENTS
PAGES
Acknowledgement
3
Liquid Media
4
Basal Media
5
Enrichment Media
7
Selective Media
9
Differential Media
13
Transport Media
17
Selective and Differential Media
19
References
20
2|Bacterial Culture Media: Types and Applications
ACKNOWLEDGEMENT I am grateful as I am managed to complete this e-book assignment with the title “Bacterial Culture Media: Types and Applications”. This e-book cannot be done without the help of my lecturers for this MLT422 course, Microbiology I, Dr Siti Nazrina Camalxaman, Madam Azlin Sham Rambely, and Madam Aziyana Zainol as they provide me the guidelines and informations on how to do this assignment. Last but not least, I would like to thank my family and friends who keep on supporting me to finish this assignment.
3|Bacterial Culture Media: Types and Applications
LIQUID MEDIA Purpose: A liquid media with lack of structure used to the production of gas, odor, changes of pH, et cetera Examples of media: 1. Trypticase Soy Broth Media 2. Nutrient Broth
▪ It allows the bacteria to move ▪ Can be used as a medium for bacteria in antibiotic susceptibility testing and disk diffusion testing Trypticase Soy Broth Media
▪ It allows the bacteria to move ▪ To cultivate many different of fastidious and non-fastidious microorganisms with non-exacting nutritional requirements.
Nutrient Broth
4|Bacterial Culture Media: Types and Applications
BASAL MEDIA Purpose: A media that is used for growth or to culture bacteria that does not require any enrichment media. It can be used to study the colony morphology of the bacteria, pigmentation and preparation for biochemical test Examples of media: 1. Nutrient Broth 2. Nutrient Agar 3. Peptone Water
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▪ It allows the bacteria to move ▪ To cultivate many different of fastidious and non-fastidious microorganisms with non-exacting nutritional requirements.
Nutrient Broth
Nutrient Agar
▪ It is a type of nutrient broth with agar powder dissolved in it ▪ Contains substances such as essential amino acid that is important for the growth of cell ▪ It can be used for isolation and identification of bacteria ▪ Can grow both gram positive and negative bacteria
▪ Used to as a bacteria growth medium ▪ Can act as abase to determine the carbohydrate fermentation pattern for non-fastidious organism ▪ Can be a based for indole production test
Peptone Water
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ENRICHMENT MEDIA Purpose: A media that is enriched as it is added with blood, serum, egg, and special extracts or vitamin that can support the growth of pathogen that requires additional nutrients Examples of media: 1. Blood Agar 2. Chocolate Agar 3. Loeffler’s Media 4. Lowenstein-Jensen Media
▪ It is considered as enrichment media because the blood act as an enrichment material ▪ The general nutrients and 5% sheep blood in the media is useful to cultivate fastidious organisms Blood Agar
▪ A heated blood agar ▪ It is suitable for growth of pneumococcus, gonococcus, meningococcus and Haemophilus spp.
Chocolate Agar
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Loeffler’s Media
▪ It contains serum that is used for enrichment ▪ Other than that, it also contains, beef extract, dextrose and proteose peptones which supply the complex nitrogenous substances and nutrients that are necessary for the growth of Corynebacterium diphtheriae.
▪ The present version of this medium has a glycerated egg-based medium formulation that can encourage the earliest possible growth of mycobacteria
Lowenstein-Jensen Media (LJ)
8|Bacterial Culture Media: Types and Applications
SELECTIVE MEDIA Purpose: A media that contain substances to favour particular bacterium by inhibiting growth of the undesired bacteria so that it allowed the desired bacteria to grow Examples of media: 1. MacConkey Agar 2. Xylose Lysine Deoxycholate (XLD) 3. Brilliant Green Agar 4. Lowenstein-Jensen Media (LJ) 5. Tellurite Media 6. Eosin-Methylene Blue Agar (EMB) 7. Hektoen Enteric Agar (HE) 8. Mannitol Salt Agar (MSA)
9|Bacterial Culture Media: Types and Applications
▪ Favours Gram-negative bacteria ▪ It contains bile salt and Crystal violet that inhibits the growth of Gram-positive bacteria
MacConkey Agar
▪ Favours Gram-negative bacteria: Salmonella spp. and Shigella spp. ▪ It contains bile salts that inhibit the growth of faecal commensal bacteria
Xylose Lysine Deoxycholate (XLD)
▪ Favours Gram-negative bacteria ▪ Use to isolate Salmonella spp. ▪ It contains dye that inhibit the growth of Gram-positive bacteria ▪ Salmonella will appear as red to pinkwhite colonies surrounded by a red zone in the medium Brilliant Green Agar
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Lowenstein-Jensen Media (LJ)
Tellurite Media
▪ A medium that incorporated with congo red and malachite green to inhibit the growth of unwanted bacteria ▪ It is used to cultivate Mycobacterium spp. particularly Mycobacterium tuberculosis ▪ M. tuberculosis will appear as brown, granular colonies
▪ It contains potassium tellurite that act as a selective agent which inhibit against most Gram-positive and Gram-negative bacteria except for Corynebacterium spp. ▪ The black deposits accumulated as Corynebacterium spp. reduces the potassium tellurite into metallic tellurium
▪ Favours Gram-negative bacteria ▪ It contains methylene blue dye that is toxic for the Gram-positive bacteria to grow
Eosin-Methylene Blue Agar (EMB)
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▪ Favours Gram-negative bacteria ▪ It contains bile salts and the dyes bromthymol blue and acid fuchsin will inhibit the growth of most Grampositive bacteria
Hektoen Enteric Agar (HE)
▪ Favours Gram-positive bacteria ▪ It contains 7.5% of sodium chloride (NaCl) which allows organism with high tolerance of salt concentration such as Staphylococcus spp.
Mannitol Salt Agar (MSA)
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DIFFERENTIAL MEDIA Purpose: A media that contains a substance that act as indicator which will change by a particular bacterium Examples of media: 1. Blood Agar 2. Eosin Methylene Blue Agar (EMB) 3. MacConkey Agar 4. Thiosulphate-Citrate-Bile salt Sucrose agar (TCBS) 5. Mannitol Salt Agar (MSA)
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Blood Agar the breaking down of blood cells in blood agar is caused by hemolysin in the organism
▪ β-hemolysis ▪ Complete lysis of red cells surrounding the colony ▪ Eg: Streptococcus pyogenes
▪ ϒ-hemolysis ▪ Red cells hemoglobin is reduced ▪ Eg: Enterococcus faecalis
▪ α-hemolysis ▪ Red cells hemoglobin is reduced ▪ Bacterium that produces hydrogen peroxide oxidise hemoglobin to green methemoglobin ▪ Eg: Streptococcus pneumoniae, Streptococcus viridans
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Eosin Methylene Blue Agar (EMB) Gram-negative bacteria that can ferment lactose will produce acid in the agar which resulting the appearance of dark purple colonies as the acid act on the dyes or sometimes dark colonies with greenish metallic sheen
▪ Coloured colonies means the organism is lactose fermenter ▪ Eg: Escherichia coli
▪ Colourless colonies means the organism is non-lactose fermenter ▪ Eg: Pseudomonas aeruginosa
MacConkey Agar It has an indicator which is neutral red that will turn red at a pH below 6.8 (acid produced) colorless at any pH greater than 6.8
▪ Non-lactose fermenter organisms do not produce acid so it appears colourless. Eg: Salmonella spp. and Shigella spp. ▪ Lactose fermenter organism produce acid so it appears pink or red. Eg: Escherichia coli
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Eosin Methylene Blue Agar (EMB) It can be used to differentiate Vibrio spp.. Organism that can ferment sucrose will acidify the medium and turn the bromthymol blue into yellow
▪ Produce flat yellow colonies ▪ Eg: Vibrio cholerae
▪ Produce colourless colonies with green centre ▪ Vibrio parahaemolyticus
Mannitol Salt Agar (MSA) Organism that can ferment mannitol will lead to producing acid and detected by phenol red indicator which helps to differentiate between Staphylococcal spp.
▪ Yellow colonies with yellow zone ▪ Staphylococcus aureus
▪ Colourless or red colonies with red zone ▪ Eg: Staphylococci other than S. aureus such as Staphylococcus epidermidis
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TRANSPORT MEDIA Purpose: A media that contain ingredients to help prevent the growth of commensals to ensure the pathogen to survive. This media is used when the specimen cannot be cultured soon when it is collected Examples of media: 1. Cary-Blair Medium 2. Amies Medium
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Cary-Blair Medium
Amies Medium
▪ It can be used to transport clinical specimens that are suspected containing enteric pathogens including Shigella, Salmonella, Vibrio cholerae, and Escherichia coli ▪ This medium has minimal nutrients and also alkaline to reduce the formation of acid, minimizing the destruction of bacteria ▪ It can be stored up to 1 year
▪ A modification of Stuart’s medium which glycerophosphate is replaced by an inorganic phosphate buffer and charcoal is added. ▪ This prolongs the viability and increase the isolation of anaerobes, delicate organism such as Neisseria gonorrhoeae, Haemophilus influenzae, Neisseria meningitidis, and Haemophilus ducreyi ▪ Suitable for 24 hours delay
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DIFFERENTIAL AND SELECTIVE MEDIA Most media that are employed in microbiological lab are both selective and differential media. It inhibits the growth of certain microorganism and differentiate colony morphologies at the same time
Some examples of media: Mannitol Salt Agar (MSA) MacConkey Agar
(EMB)
▪ Differentiate Staphylococcus aureus ▪ High salt concentration inhibits most organisms to grow ▪ Differentiate lactose fermenter and nonlactose fermenter bacteria ▪ Favours Gram-negative bacteria and inhibit Gram-positive bacteria ▪ Differentiate sucrose fermenter and nonsucrose fermenter bacteria ▪ Favours Gram-negative bacteria and inhibit Gram-positive bacteria
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REFERENCES
Aryal. S. (2018, October 26). Blood Agar- Composition, Preparation, Uses and Pictures. Retrieved from https://microbiologyinfo.com/blood-agar-compositionpreparation-uses-and-pictures/
Aryal. S. (2018, October 26). List of culture media used in microbiology with their uses. Retrieved from https://microbiologyinfo.com/list-of-culture-media-used-inmicrobiology-with-their-uses/ Aryal. S. (2018, September 27). Nutrient Agar: Composition, Preparation and Uses. Retrieved from https://microbiologyinfo.com/nutrient-agar-compositionpreparation-anduses/#:~:text=Nutrient%20Agar%20is%20a%20general,need ed%20for%20the%20bacterial%20growth. Fattah. S. (2015, November 25). Types of culture media. Retrieved from https://www.slideshare.net/HiwrHastear/types-ofculture-media Quinlan. S. (2018, April 26). Types of Agar Plates. Retrieved from https://sciencing.com/types-agar-plates-8131230.html Tankeshwar. A. (2010, July 24). Bacterial Culture Media: classification, types and uses. Retrieved from https://microbeonline.com/types-of-bacteriological-culturemedium/ Types of culture media used in microbiology. Retrieved from https://www.aladdine.com/up_files/docs/Types%20of%20culture%20media%20us ed%20in%20microbiology.pdf
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