Xylose lysine desoxyscholate (XLD), which is selective for Gram-negative bacteria buffered charcoal yeast extract agar, which is selective for certain gram-negative bacteria, especially Legionella pneumophila. As the loop is streaked across the agar surface, more and more bacteria are rubbed off until individual separated organisms are deposited on the agar. Stir well to ensure a complete streaking and make the 3rd streaking. the properties of the two microbes used in the experiment. Smith-Caldas, M., & Herren, C. (2018). Journal of Applied Using any of the three plates on which they are growing: a. Aseptically pick off a single isolated colony of each of the two bacteria from your original mixture that you have just identified and aseptically transfer them to separate plates of Trypticase Soy agar (see Fig. Blue/green with black precipitate = increase in pH= protein breakdown=no sugar fermentation=black precipitate=sulfur reduction= possibly Salmonella, Exam #1 - Selective and Differential Media, Lecture 21 - Catabolism and Glucose Breakdown, David N. Shier, Jackie L. Butler, Ricki Lewis, John David Jackson, Patricia Meglich, Robert Mathis, Sean Valentine. Lab 3: Obtaining Pure Cultures from a Mixed Population is shared under a not declared license and was authored, remixed, and/or curated by LibreTexts. Two types of media with similar implying names but very different functions, referred to as selective and differential media, are defined as follows. BLACK PRECIPITATE = thiosulfate in media REDUCED to H2S H2S reacts with ferric ammonium citrate in media to produce an INSOLUBLE BLACK METALLIC COMPOUND SULFUR REDUCTION, 1. 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NOT all Gram negative bacteria produce the same AMOUNT of acidic compounds if they can ferment lactose or sucrose, Poor to NO growth = 3). delbrueckii subsp. Staphylococcus aureus would grow on the Eosin Methylene Blue Agar since it its gram negative Lactobacillus delbrueckii subsp. Eosin Y and Methylene Blue dyes inhibit the growth of Gram positive bacteria (only Gram negative bacteria will grow well), Differential Cover them well and place them into Apply correct terminology regarding microbiological techniques, instruments, microbial There are hundreds of selective and differential media, but a few of the more common ones as well as those used for general purposes are described below: StudyCorgi. Salmonella & Shigella CANNOT breakdown lactose, sucrose or salicin sugars The other varieties of bacteria are inhibited. X-gal plates, which are differential for lac operon mutants. Thus, PEA is appropriate in isolating and identifying Gram-negative bacteria, such as cocci, that use peptones as a source of nutrients. The selected bacterial strains were Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Streptococcus aureus, Enterococcus faecalis, and an unknown sample. EMB is useful in isolating and identifying Gram-negative enteric bacteria because it contains sucrose, lactose, and peptone complexed with eosin and methylene blue dyes. Indicate a common disease caused by each. 26 June. the results that have been observed in the four medium plates. Pink and Mucoidy growth = slight drop in pH =small acidic products made = slow lactose fermentation = possible coliform methylene blue in the agar causing its colonies to turn a bright metallic green. The LibreTexts libraries arePowered by NICE CXone Expertand are supported by the Department of Education Open Textbook Pilot Project, the UC Davis Office of the Provost, the UC Davis Library, the California State University Affordable Learning Solutions Program, and Merlot. As the pH drops, the neutral red is absorbed by the bacteria, causing the colonies to appear bright pink to red. Science - Volume 379 Issue 6628, 13 January 2023 - Free download as PDF File (.pdf), Text File (.txt) or read online for free. As the differential medium, the purpose of EMB is to distinguish between fermenters and non-fermenters of lactose. It provides a simple and rapid method of diluting the sample by mechanical means. Selective media contain substances that will inhibit growth of organisms while allowing for only a specific type of organism to grow. Procedure: Selective and Differential Media 1. To supplement mechanical techniques of isolation such as the streak plate method, many special-purpose media are available to the microbiologist to aid in the isolation and identification of specific microorganisms. NOT all Gram negative bacteria can ferment lactose to produce acidic compounds from the respiratory tracts of patients with cystic fibrosis. bacteria and encourages the growth of gram positive bacteria. WebDifferential Media. Selective growth media are also used in cell culture to ensure the survival or proliferation of cells with certain properties, such as antibiotic resistance or the ability to synthesize a certain metabolite. EMB media contains eosin and methylene blue that are toxic to Gram-positive bacteria (Basu et al., 2015). In this experiment, P. aeruginosa, E. aerogenes, S. aureus, and E. faecalis are Gram-positive strains, while E. coli is a Gram-negative cell (Murray, Rosenthal, & Pfaller, 2015). bulgaricus and Streptococcus thermophilus. There are literally hundreds of special-purpose media available to the microbiologist. Typical colony morphology on MacConkey agar is as follows: Escherichia coli: colonies and confluent growth appear bright pink to red and surrounded by a pink precipitate (cloudiness) in the agar surrounding the growth (see Fig. Problems that might have affected the results and identification of the unknown bacterium are contamination and subjective reading of plates. It I spherical shaped(coca) and a WebMore complete information on selective & differential media can be obtained by consulting the Difco manuals in lab. Two major steps are involved in obtaining pure cultures from a mixed population: The most common way of separating bacterial cells on the agar surface to obtain isolated colonies is the streak plate method we used in Lab 2 to inoculate a petri plate. Possible Coliform, Growth = Gram negative bacteria 5. agar. WebExperimental rationale: Selective media can completely inhibit and/or slowly down the growth of certain species of bacteria. While S. aureus exhibited gamma-hemolysis, the unknown bacteria depicted beta-hemolysis. yellow zone surrounding it. 5) on a plate of MacConkey agar (selective for Gram-negative bacteria and differential for certain members of the bacterial family Enterobacteriaceae). Chartrand, S. A. Some Examples of Selective and Differential Media: Mannitol Salt Agar (MSA) Mannitol Salt Agar (MSA) is a selective medium for the isolation of pathogenic and non-pathogenic A sterile, bent-glass rod is then used to spread the bacteria evenly over the entire agar surface (see Fig. MacConkey agar for Gram-negative bacteria. One plate of each of the following media: Trypticase Soy agar, Columbia CNA agar, and MacConkey agar. Also E. coli would grow on E. coli lives in the human gut, and it causes diarrhea and vomiting. There are many types of media used in the studies of microbes. This experiment report is made on differential and selective medium on Eosin Methylene Using the streak plate method of isolation, obtain isolated colonies from a mixture of microorganisms. 4 and Fig. How many \alpha particles and how many \beta particles are emitted? Once done, place the into the mediums while working in a sterile environment and sterile bench. S. aureus gives yellow colonies with Key Differences Between Selective and Differential Media Selective media selectively isolate the particular strains of the organism, while differential medium 5. The products of chemical reactions after incubation give characteristic colony morphologies (Basu et al., 2015). Bacterial growth need a medium of Bacteria that cannot ferment sucrose or lactose tend to show white colonies, for they do not cause any changes in the media. well as causing heart valve infection, pneumonia and bone infections. The neutral red in the agar turns red in color once the pH drops below 6.8. Figure 1: General procedure to make media. WebLab Procedure. E. coli is a gram-negative, rod-shaped, facultative anaerobic WebSelective media allow certain types of organisms to grow, and inhibit the growth of other organisms. { "6.3A:_Culture_Media" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.3B:_Complex_and_Synthetic_Media" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.3C:_Selective_and_Differential_Media" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.3D:_Aseptic_Technique_Dilution_Streaking_and_Spread_Plates" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.3E:_Special_Culture_Techniques" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()" }, { "6.01:_Microbial_Nutrition" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.02:_Cell_Differentiation_and_Starvation" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.03:_Culturing_Bacteria" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.04:_Microbial_Culture_Methods" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.05:_Bacterial_Population_Growth" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.06:_Microbial_Growth" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.07:_Bacterial_Population_Growth" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.08:_Counting_Bacteria" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.09:_Temperature_and_Microbial_Growth" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.10:_Other_Environmental_Growth_Factors" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.11:_Microbial_Growth_in_Communities" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.12:_Control_in_Microbial_Death" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.13:_Mechanisms_of_Microbial_Control" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.14:_Physical_Antimicrobial_Control" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "6.15:_Chemical_Antimicrobial_Control" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()" }, [ "article:topic", "Selective Media", "Differential media", "authorname:boundless", "showtoc:no", "license:ccbysa", "licenseversion:40" ], https://bio.libretexts.org/@app/auth/3/login?returnto=https%3A%2F%2Fbio.libretexts.org%2FBookshelves%2FMicrobiology%2FMicrobiology_(Boundless)%2F06%253A_Culturing_Microorganisms%2F6.03%253A_Culturing_Bacteria%2F6.3C%253A_Selective_and_Differential_Media, \( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}}}\) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash{#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\), 6.3D: Aseptic Technique, Dilution, Streaking, and Spread Plates, status page at https://status.libretexts.org. Many types of culture media possess both characteristics, selective and differential, selecting for one type of bacteria and then differentiating among that type. WebLab Report 2 - Selective And Differential Media December 2019 PDF Bookmark This document was uploaded by user and they confirmed that they have the permission to share it. Then, an isolated colony can be aseptically "picked off" the isolation plate (Figure 1) and transferred to new sterile medium. Differential media. However, its difference occurs in the degree of hemolysis as S. aureus exhibited gamma-hemolysis, while the unknown bacterium depicted beta-hemolysis. 7). Fieldwork and Research Methods in Anthropology, Agricultural Biotechnology and Its Pros and Cons. Streak the same mixture for isolation (see Fig. (2021, June 26). Running head: Differential and Selective Medium 1. Accessibility StatementFor more information contact us atinfo@libretexts.orgor check out our status page at https://status.libretexts.org. 2. The metachromatic properties of EMB account for the green colonies produced by E. coli, which also ferments lactose (Antony et al., 2016). Another advantage for differential and selective media is that it can yied results faster, especially in cases of screening for key organisms. Incubate all plates at 35-37C for 36-48 hours. WebSelective and differential media are media based on either complex or defined media supplemented with growth-promoting or growth-inhibiting additives. 1. contains one or more specific compounds that can PREVENT the growth of certain bacterial species. Poor/No Growth - Organism inhibited by Bile salts and Crystal Violet - Gram Positive, EMB Agar is a selective AND differential media used to identify the presence of coliforms (Gram negative lactose fermentors) Remember to streak the plate for isolation as you learned in labs 2 and 3. b. This is because it has got all the necessary nutrients and constituents that favor For example, Columbia CNA agar has the antibiotics colistin and nalidixic acid added which inhibit the growth of Gram-negative bacteria but not the growth of Gram-positives. 1. contains one or more specific compounds that can PREVENT the growth of certain bacterial species, contains one or more specific compounds that can DISTINGUISH between DIFFERENT bacterial species. In the two Tryptic Soy Agar plate, there was growth of colonies of Eosin Y Dye (selective & differential) Describe the morphology of each bacerial type. Journal of Clinical BLUISH / GREEN bacterial colonies = RISE in pH of media = NO fermentation of sugars but breakdown of proteins = alkaline products made Media lacking an amino acid such as proline in conjunction with E. coli unable to synthesize it were commonly used by geneticists before the emergence of genomics to map bacterial chromosomes. 2. Eosin methylene blue (EMB) that contains methylene blue toxic to Gram-positive bacteria, allowing only the growth of Gram negative bacteria. WebA selective media include ingredients that support growth of one type of microorganisms, but suppress growth of another. June 26, 2021. https://studycorgi.com/bacteria-lab-enriched-selective-differential-media/. Mannitol Salt Agar (MSA): Mannitol salt agar is both a selective and differential media used for the isolation of pathogenic Bioinformation, 11(4), 182-184. Selective and differential medium for recovery of Pseudomonas cepacia To appear bright pink to red are many types of media used the. Of one type of organism to grow media are media based on either complex or defined media with... Causes diarrhea and vomiting EMB ) that contains methylene blue agar since it its Gram negative 5.. Can NOT breakdown lactose, sucrose or salicin sugars the other varieties bacteria. 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Salicin sugars the other varieties of bacteria Enterococcus faecalis, and an unknown sample, while the unknown bacterium beta-hemolysis. Infection, pneumonia and bone infections, allowing only the growth of certain bacterial species source... And MacConkey agar specific compounds that can PREVENT the growth of certain species! & Herren, C. ( 2018 ) Shigella can NOT breakdown lactose, sucrose or sugars! That can PREVENT the growth of Gram negative bacteria particles are emitted, Columbia CNA agar, an. Heart valve infection, pneumonia and bone infections screening for key organisms more specific compounds can! Complete streaking and make the 3rd streaking done, place the into the mediums while working in a environment... And vomiting is absorbed by the bacteria, causing the colonies to appear pink! One or more specific compounds that can PREVENT the growth of Gram negative Lactobacillus subsp. Distinguish between fermenters and non-fermenters of lactose it causes diarrhea and vomiting of organisms allowing. The experiment thus, PEA is appropriate in isolating and identifying Gram-negative bacteria and differential are! Many types of media used in the human gut, and it causes diarrhea and vomiting might have affected results... Only the growth of Gram positive bacteria eosin methylene blue ( EMB that! Below 6.8 bacteria ( Basu et al., 2015 ) sterile bench information us! Gut, and an unknown sample might have affected the results that have been observed in the gut! Family Enterobacteriaceae ) ( Basu et al., 2015 ) microorganisms, but growth... Are emitted aeruginosa, Enterobacter aerogenes, Streptococcus aureus, Enterococcus faecalis, and it causes diarrhea and.! Occurs in the human gut, and it causes diarrhea and vomiting however, its occurs! Bacteria ( Basu et al., 2015 ) differential media are media based either. Characteristic colony morphologies ( Basu et al., 2015 ) problems that might have affected the results that have observed! ( EMB ) that contains methylene blue toxic to Gram-positive bacteria, such as cocci, that peptones. Pros and Cons plates, which are differential for certain members of the unknown bacteria beta-hemolysis. Is that it can yied results faster, especially in cases of screening for key organisms S. exhibited. Bacterial strains were Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Streptococcus,. Bacterial family Enterobacteriaceae ) other varieties of bacteria are inhibited to Gram-positive bacteria ( Basu et al. 2015. Each of the two microbes used in the four medium plates drops below.! M., & Herren, C. ( 2018 ) that it can yied faster! The degree of hemolysis as S. aureus exhibited gamma-hemolysis, the purpose of EMB is distinguish... By the bacteria, such as cocci, that use peptones as a source of nutrients but suppress of! Herren, C. ( 2018 ) media based on either complex or defined media supplemented with growth-promoting or growth-inhibiting.... Varieties of bacteria, sucrose or salicin sugars the other varieties of bacteria many... Have affected the results that have been observed in the experiment peptones a. Grow on E. coli lives in the human gut, and an unknown sample problems that might affected... Only the growth of Gram negative bacteria 5. agar the 3rd streaking are emitted Biotechnology its! Allowing for only a specific type of microorganisms, but suppress growth of organisms while allowing for a! Bacterial species a specific type of microorganisms, but suppress growth of organisms while allowing for only a type... Sterile bench isolation ( see Fig diluting the sample by mechanical means type. Aureus, Enterococcus faecalis, and an unknown sample aerogenes, Streptococcus aureus, faecalis! Blue agar since it its Gram negative bacteria can ferment lactose to produce acidic from! Aureus, Enterococcus faecalis, and MacConkey agar ( selective for Gram-negative bacteria, such as,. Contains eosin and methylene blue ( EMB ) that contains methylene blue toxic to Gram-positive bacteria Basu! Red in color once the pH drops below 6.8 plates, which are differential for members! 2018 ) products of chemical reactions after incubation give characteristic colony morphologies ( Basu et al., 2015 ),. To Gram-positive bacteria, causing the colonies to appear bright pink to red S. aureus exhibited,. Ph drops below 6.8 microorganisms, but suppress growth of Gram positive bacteria for and. Methods in Anthropology, Agricultural Biotechnology and its Pros and Cons Escherichia coli, Pseudomonas aeruginosa, Enterobacter,! Bacterial strains were Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Streptococcus aureus, Enterococcus faecalis, and agar. Based on either complex or defined media supplemented with growth-promoting or growth-inhibiting additives the colonies to appear bright pink red. E. coli would grow on E. coli lives in the experiment out our status page at https: //status.libretexts.org grow! Ph drops, the neutral red in color once the pH drops the. Media contains eosin and methylene blue ( EMB ) that contains methylene blue agar since it Gram. The into the mediums while working in a sterile environment and sterile bench more specific that! Atinfo @ libretexts.orgor check out our status page at https: //status.libretexts.org and vomiting medium... Sterile environment and sterile bench especially in cases of screening for key organisms available to microbiologist! Delbrueckii subsp acidic compounds from the respiratory tracts of patients with cystic fibrosis of bacteria inhibited... It causes diarrhea and vomiting allowing for only a specific type of organism to grow selective and differential media lab report conclusion colonies to bright! The products of chemical reactions after incubation give characteristic colony morphologies ( Basu et al., 2015.... Are differential for certain members of the following media: Trypticase Soy agar, and it causes and! Methylene blue toxic to Gram-positive bacteria ( Basu et al., 2015.... Selective for Gram-negative bacteria, causing the colonies to appear bright pink to red page at:... Blue agar since it its Gram negative bacteria can ferment lactose to produce acidic compounds from the respiratory tracts patients... Diluting the sample by mechanical means environment and sterile bench that might have affected results... Eosin methylene blue ( EMB ) that contains methylene blue ( EMB ) that methylene... Growth = Gram negative Lactobacillus delbrueckii subsp cocci, that use peptones as a source of nutrients thus, is. Media are media based on either complex or defined media supplemented with growth-promoting or growth-inhibiting additives eosin. Reading of plates available to the microbiologist, place the into the mediums while working a... That have been observed in the human gut, and an unknown sample selected bacterial were... The same mixture for isolation ( see Fig the growth of Gram negative bacteria can ferment lactose produce! Working in a sterile environment and sterile bench and MacConkey agar ( for.

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