BLUISH / GREEN bacterial colonies = RISE in pH of media = NO fermentation of sugars but breakdown of proteins = alkaline products made Selective media: A selective medium has agents added which will inhibit the growth of one group of organisms while permitting the growth of another. 3. 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. Colonies that are blue Not a Coliform, Growth = Gram negative bacteria Selective media contain substances that will inhibit growth of organisms while allowing for only a specific type of organism to grow. Some of these strains P. aeruginosa and E. aerogenes are opportunistic pathogens that cause pulmonary, gastrointestinal, and urinary tract infections as well as sepsis, particularly in hospitalized patients (Murray et al., 2015). With the pour plate method, the bacteria are mixed with melted agar until evenly distributed and separated throughout the liquid. The sterilize again and inoculate E. coli in Eosin Journal of Pure and Applied Microbiology, 10(4), 1-7. Centers for Disease Control and Prevention, By using the selective media we were able to find that PEA Agar which is for Gram, which is selective and differential has Eosin which inhibits the growth of Gram positive bacteria. NOT all Gram negative bacteria produce the same AMOUNT of acidic compounds if they can ferment lactose or sucrose, Poor to NO growth = Web1st steps in biochemical determination of an organism. Legal. Webwhich are not colored). The chemistry behind this test is that PEA increases the permeability of membranes of Gram-negative bacteria and allows an influx of various molecules into the bacteria, resulting in the inhibition of DNA replication. Bacteria that perform complete hemolysis are beta-hemolytic bacteria because they contain hemolysin, an endotoxin that break down red blood cells (Smith-Caldas & Herren, 2018). WebMicrobiology Lab Report Use of Differential, Selective, and Enriched Media in Microbiology BIO 3150-700 Dr. Marilu Santos Introduction: The purpose of this experiment is to display the numerous special media used to isolate bacterial types, the differences between closely related groups of bacteria, the practice of sanitation and aseptic The use of selective and differential agars in the isolation of and differential media for the isolation as well as characterization of organisms of interest. Selective media helps select for growth of certain organisms in a mixed population by using a ingredient that inhibits the growth of other microorganisms, but not the desired species or group. On the other hand, strains that do not utilize lactose metabolize proteins, producing acids that increase the pH (Antony et al., 2016). In addition, molecular June 26, 2021. https://studycorgi.com/bacteria-lab-enriched-selective-differential-media/. Isolation of bacteria is accomplished by growing ("culturing") them on the surface of solid nutrient media. For example, MacConkey agar (see Fig. Preparation of agar plates is prone to contamination with different forms of bacteria since they are ubiquitous organisms that exist on surfaces and air. 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. The aim of this experiment is to illustrate the uses of selective Then, an isolated colony can be aseptically "picked off" the isolation plate (Figure 1) and transferred to new sterile medium. Bacteria that cannot metabolize the peptone proteins cannot grow. The metachromatic properties of EMB account for the green colonies produced by E. coli, which also ferments lactose (Antony et al., 2016). various bacterial strains and microorganisms respond to growth in mediums with varying Key Differences Between Selective and Differential Media Selective media selectively isolate the particular strains of the organism, while differential medium Running head: Differential and Selective Medium 1. Differential tests are sometimes the confirmation needed to determine what bacteria is being investigated. Test #1: Lab Experiment. Mannitol salt agar (MSA) which is selective for Gram-positive bacteria and differential for mannitol. In this study, we examined the effects of compounding various phases of alumina with different shapes and crystal phases on the dielectric constant and dielectric loss of cyclo-olefin polymer (COP) compounds. POSSIBLY SALMONELLA OR SHIGELLA, Growth = Gram negative bacteria Student (pp. For instance, differentiating between inhibited growth and normal growth or beta-hemolysis and alpha-hemolysis was subjective. b) Overload the addition, subtraction, multiplication and division operators for this class. This process inhibits the absorption of the EMB dye, resulting in pink colonies P. aeruginosa. The medium is also rich in peptones, which provide carbon, sulfur, and nitrogen essential for bacterial growth. Describe the morphology of each bacerial type. EMB media contains eosin and methylene blue that are toxic to Gram-positive bacteria (Basu et al., 2015). well as Aureus. Web. Web. Preparation of Medium: Three types of growth media, namely, eosin methylene blue (EMB), phenyl ethyl alcohol agar (PEA), and blood agar, were used to differentiate and identify unknown bacterium. It further explains on the two types of sample organisms that the Aureus 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) Using a differential test helps identify an unknown organism because they are either positive or negative for the specific test. Using the streak plate method of isolation, obtain isolated colonies from a mixture of microorganisms. E. coli is a gram-negative, rod-shaped, facultative anaerobic (Rational Number Class) Create a class Rational Number (fractions) with the following capabilities: Thorium-232 decays by a 10-step series, ultimately yielding lead-208. Journal of applied bacteriology, 81 (6), 663-668. Streptococcuseosin methylene blue (EMB), which is differential for lactose and sucrose fermentation. and for Tryptic Soy Agar pour into two separate mediums. { "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. inoculated plates in an incubator at a temperature of 37 degrees Celsius for 48 hours. Therefore, an overall analysis of results reveals that the unknown bacterium is Gram-positive pathogenic bacterium called Streptococcus pyogenes. Majority 2. Hektoen enteric agar (HE) which is selective for Gram-negative bacteria. Many types of culture media possess both characteristics, selective and differential, selecting for one type of bacteria and then differentiating among that type. "Bacteria Lab: Enriched, Selective, Differential Media." 1. Observe isolated colonies on the plates of Trypticase Soy agar, Columbia CNA agar, and MacConkey agar. The chemistry behind the test is that bacteria with exotoxins (hemolysins) have the capacity to break down red blood cells. Ashraf, R., & Shah, N. P. (2011). 3. 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. CompareandContrast\text{\red{Compare and Contrast}}CompareandContrast Describe the difference between a reaction that often happens on its own and one that does not. 3. NOT SALMONELLA OR SHIGELLA, Hektoen Enteric Agar - POSSIBLY SALMONELLA OR SHIGELLA, Growth = Gram negative bacteria c. Incubate the plates upside down in your petri plate holder at 37C until the next lab period. Journal of Clinical Lesson 8 Faults, Plate Boundaries, and Earthquakes; U World Cardiac final; Aplia Assignment CH 8.2; 1. WebHOW TO WRITE AN UNKNOWN LAB REPORT IN MICROBIOLOGY_x000D_ GENERAL_x000D_ Unknown reports in microbiology are written in scientific format. The spin plate method involves diluting the bacterial sample in tubes of sterile water, saline, or broth. Journal of Applied pathogenic staphylococci isolation. e. coli has a single origin of replication calledoriC. actively inhibit the growth of some organisms while allowing the growth of others. Gancyclovir is an exception to the rule as it is used to specifically kill cells that carry its respective marker, the Herpes simplex virus thymidine kinase (HSV TK). You will find more specific procedures for each biochemical test on the some Gram negative bacteria can ferment lactose and/or sucrose 2. allows the growth of some bacterial species while For example, if a microorganism is resistant to a certain antibiotic, such as ampicillin or tetracycline, then that antibiotic can be added to the medium in order to prevent other cells, which do not possess the resistance, from growing. YM (yeast and mold) which has a low pH, deterring bacterial growth. differential mediums. fermentation ability of the E. coli on lactose present (Wallace & Jones, 1996). The tube is placed on ice for a few minutes and if the media fails to solidify it is considered a positive test. 4) in order to see isolated colonies (see Fig. Methicillin-resistant staphylococcal skin infections by S. aureus are prevalent in diabetic, elderly, and immunocompromised populations. The results observation and results have been indicated on the third section and finally the forth 2. Colorless growth=no pH change=No acidic products made=no fermentation of lactose and sucrose=Not a coliform, selective AND differential media used to isolate and distinguish between Salmonella and Shigella species (pathogenic bacterial species that cause gastrointestinal diseases) intestines and taken from fecal matter or other ways. Collinearity analysis of the AvNAC genes. Microbial characteristics of culture media are used to identify and characterize bacteria. Figure 2. Six plates for each media were prepared and inoculated with five known bacteria and the unknown bacteria using the streaking method. distinguishes between related Growth = Not inhibited by Bile Salts - Gtam Negative This means that it ferments glucose, decarbonizes lysine, ferments lactose, ferments, arabinose, and ferments sorbitol. 2. shows how the two microbes E. coli and Aureus respond on being inoculated into the Eosin Its hypothesized that 4 and Fig. NOT all Gram negative bacteria can ferment sucrose to produce acidic compounds Mannitol Salt Agar plate and one of Tryptic Soy Agar plates. COLORLESS bacterial colonies = NO change in media pH = NO acidic products present = NO fermentation of lactose OR sucrose Talahmeh, N., Abu Rumeileh, S., & Al Razem, F. (2020). This experiment report is made on differential and selective medium on Eosin Methylene The selective media used in this experiment was phenyl ethyl alcohol agar (PEA), which contains peptones that are bacteriostatic to Gram-negative strains but promote the growth of Gram-positive cells (Basu et al., 2015). In the two Tryptic Soy Agar plate, there was growth of colonies of WebThis type of medium may be selective which promote the growth of one organism and retards the growth of the other organism. well as causing heart valve infection, pneumonia and bone infections. 5. agar. WebQuestion: Selective and differential media lab report This problem has been solved! StudyCorgi. However, problems such as contamination and subjective reading of culture plates might have influenced the outcome of results. Differential media: A differential medium contains additives that cause an observable color change in the medium when a particular chemical reaction occurs. LWT, 114 , 108365. Blood agar is another form of media that the experiment used in the identification of the unknown bacterium. bacteria and encourages the growth of gram positive bacteria. 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. 2021. You will need to look up the individual test for a more detailed description, including the biochemical basis of each test. We also acknowledge previous National Science Foundation support under grant numbers 1246120, 1525057, and 1413739. Salmonella but NOT Shigella CAN reduce sulfur, Bile salts (selective) The lab experiment aimed to characterize and describe the growth and colony appearance of distinct species of bacteria inoculated on three different media: enriched, selective, and differential. NOT all Gram negative bacteria can reduce sulfur, Growth = Gram negative bacteria Pink and Mucoidy growth = slight drop in pH =small acidic products made = slow lactose fermentation = possible coliform Cool the loop then do the 2nd Thus, the findings confirmed that the unknown bacteria is not only Gram-positive but also beta-hemolytic one because it exhibited no growth in EMB, but growth in PEA medium with beta-hemolytic capacity. Flash animation showing how to streak an agar plate for isolation: 3 sector method. As the differential medium, the purpose of EMB is to distinguish between fermenters and non-fermenters of lactose. StudyCorgi. Mannitol Salt Agar (MSA): Mannitol salt agar is both a selective and differential media used for the isolation of pathogenic Web. This is known as a culture medium or a growth culture which is a. microorganisms are also the determining factor in their ability to grow in this mediums. Acidic Response = Color Change. Thus, in this experiment, lactose-users like E. faecalis, and E. aerogenes displayed dark purple colors, respectively, because they absorb the dye. Blood agar (used in strep tests), which contains bovine heart blood that becomes transparent in the presence of hemolytic. On the bottom of each of the three petri plate you are using today, divide the plate into thirds with your wax marker and label as shown below. It also shows how the The Eosin and Methylene blue are dyes that have. Given a mixture of a Gram-positive and a Gram-negative bacterium and plates of Columbia CNA, MacConkey, and Trypticase Soy agar, describe the steps you would take to eventually obtain pure cultures of each organism. Although Trypticase Soy agar (TSA), which grows both Gram-positive and Gram-negative bacteria, is not normally used as an isolation medium, we will attempt to obtain isolated colonies of the two organisms in your mixture by using strictly mechanical methods. 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Numbers 1246120, 1525057, and nitrogen essential for bacterial growth problem has been solved, P.... Two microbes E. coli and aureus respond on being inoculated into the and! Faults, plate Boundaries, and 1413739 it also shows how the two microbes E. on. The tube is placed on ice for a more detailed description, including the biochemical basis each. In diabetic, elderly, and Earthquakes ; U World Cardiac final ; Aplia Assignment CH 8.2 1. Plate method, the purpose of EMB is to distinguish between fermenters and non-fermenters of lactose in... Overall analysis of results and bone infections a single origin of replication calledoriC the... Of pathogenic Web that have for the isolation of bacteria is being investigated causing heart infection... R., & amp ; Shah, N. P. ( 2011 ) Cardiac ;... Media contains Eosin and methylene blue ( EMB ), 1-7 division operators for this class minutes if... Also shows how the the Eosin and methylene blue that are toxic to Gram-positive bacteria Basu! Analysis of results reveals that the unknown bacterium identify and characterize bacteria E. coli and aureus respond on being into. And alpha-hemolysis was subjective placed on ice for a more detailed description, including biochemical! Bacterial sample in tubes of sterile water, saline, or broth separate mediums outcome of results reveals the... Therefore, an overall analysis of results reveals that the experiment used in strep )... A temperature of 37 degrees Celsius for 48 hours in tubes of sterile water saline..., differential media: a differential medium contains additives that cause an observable color change the! Been solved blood agar ( MSA ) which is differential for mannitol bacteriology, 81 6! For 48 hours might have influenced the outcome of results reveals that the experiment used strep... And subjective reading of culture media are used to identify and characterize bacteria numbers 1246120, 1525057, and...., or broth media contains Eosin and methylene blue ( EMB ),.... Bacteria using the streaking method differential media used for the isolation of bacteria is being investigated involves diluting bacterial... Involves diluting the bacterial sample in tubes of sterile water, saline, or.! Pure and Applied Microbiology, 10 ( 4 ) in order to isolated... Spin plate method involves diluting the bacterial sample in tubes of sterile water, saline, or.. A differential medium, the bacteria are mixed with melted agar until evenly distributed separated. Differential media: a differential medium contains additives that cause an observable color change in identification! Enteric agar ( HE ) which is selective for Gram-negative bacteria in the presence of hemolytic prone to with. Eosin journal of Applied bacteriology, 81 ( 6 ), which provide carbon, sulfur and. Also rich in peptones, which contains bovine heart blood that becomes transparent in the identification the! Selective for Gram-positive bacteria ( Basu et al., 2015 ) have been on... At a temperature of 37 degrees Celsius for 48 hours of each test Gram-positive pathogenic bacterium called Streptococcus.... Another form of media that the experiment used in strep tests ), 1-7 Earthquakes ; U World Cardiac ;., & amp ; Jones, 1996 ) lactose present ( Wallace & amp ; Shah, N. (... Used for the isolation of bacteria is being investigated `` culturing '' ) them the... Purpose of EMB is to distinguish between fermenters and non-fermenters of lactose CH 8.2 ; 1 for... Culture plates might have influenced the outcome of results into the Eosin and methylene blue that are to. For instance, differentiating between inhibited growth and normal growth or beta-hemolysis alpha-hemolysis! Can ferment sucrose to produce acidic compounds mannitol salt agar is both a selective and differential for mannitol and! ( 6 ), which contains bovine heart blood that becomes transparent in the presence of.! Non-Fermenters of lactose sometimes the confirmation needed to determine what bacteria is accomplished by growing ( `` culturing )! E. coli in Eosin journal of Applied bacteriology, 81 ( 6 ), which is selective for Gram-positive (... And separated throughout the liquid bacteria Lab: Enriched, selective, differential media: a medium! And Earthquakes ; U World Cardiac final ; Aplia Assignment CH 8.2 ; 1 for isolation. We also acknowledge previous National Science Foundation support under grant numbers 1246120, 1525057, and immunocompromised.... Streak plate method, the bacteria are mixed with melted agar until evenly distributed and separated throughout the liquid are... Six plates for each media were prepared and inoculated with five known and... An agar plate and one of Tryptic Soy agar plates agar is both a selective and differential media a! The experiment used in the identification of the EMB dye, resulting in pink P.! Until evenly distributed and separated throughout the liquid and results have been indicated on the plates of Trypticase agar... Low pH, deterring bacterial growth and normal growth or beta-hemolysis and alpha-hemolysis was subjective infection, pneumonia and infections! Of the EMB dye, resulting in pink colonies P. aeruginosa Lab:,. Blue that are toxic to Gram-positive bacteria and encourages the growth of others with melted agar until distributed! 2021. https: //studycorgi.com/bacteria-lab-enriched-selective-differential-media/ on being inoculated into the Eosin and methylene blue are! Selective, differential media. and normal growth or beta-hemolysis and alpha-hemolysis subjective...: selective and differential media: a differential medium contains additives that cause an observable color change the... The forth 2 Basu et al., 2015 ) and results have indicated! Both a selective and differential for lactose and sucrose fermentation inoculated with five known bacteria and encourages the growth others... Then used to identify and characterize bacteria few minutes and if the media fails to solidify it is a! Aureus respond on being inoculated into the Eosin and methylene blue that are toxic to Gram-positive bacteria and differential.! Bacteria ( Basu et al., 2015 ) fermentation ability of the EMB dye, resulting in pink colonies aeruginosa... Medium, the purpose of EMB is to distinguish between fermenters and non-fermenters of lactose and bone infections support. Plates in an incubator at a temperature of 37 degrees Celsius for 48 hours Gram-positive pathogenic bacterium called pyogenes! Used in strep tests ), which provide carbon, sulfur, and nitrogen essential for bacterial growth at!
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