A. This plate is called an isolation plate. and inorganic salts, hardened by the addition of 1.5% agar. Sodium thiosulfate (sulfur source) Therefore, an overall analysis of results reveals that the unknown bacterium is Gram-positive pathogenic bacterium called Streptococcus pyogenes. Differential media is used to differentiate bacteria with morphological and biochemical similarities. Differential media. Eosin Y and Methylene Blue dyes inhibit the growth of Gram positive bacteria (only Gram negative bacteria will grow well), Differential Describe the morphology of each bacerial type. Web. pathogenic produces small yellow colonies within a yellow region. 4. If you are the original creator of this paper and no longer wish to have it published on StudyCorgi, request the removal. Today we will combine both a mechanical isolation technique (the streak plate) with selective and selective-differential media to obtain pure cultures from a mixture of bacteria. P. aeruginosa, E. aerogenes, and E. coli are rod-shaped microorganisms (Murray et al., 2015). Procedure: Selective and Differential Media 1. In the Eosin Methylene Blue Agar plate, E. coli appears large. NOT all Gram negative bacteria can ferment sucrose to produce acidic compounds 8). Bacteria Lab: Enriched, Selective, Differential Media. If you are author or own the copyright of this book, please report to us by using this DMCA report form. Acidic Response = Color Change. YELLOW MEDIA CHANGE= + fermentation of mannitol --> acid. Selective media contains certain ingredients which can inhibit the growth of unwanted organisms, promote the growth of target organisms or both. shows how the two microbes E. coli and Aureus respond on being inoculated into the Eosin Figure 1: General procedure to make media. nature is due to the dye properties of the stains. high [NaCl] of 7.5%. 3). Evolution of bacterial and fungal growth media. Webwhich are not colored). The tube is placed on ice for a few minutes and if the media fails to solidify it is considered a positive test. In Food Microbiology Laboratory for the Food Science staphylococci which are halophilic bacteria. In the two Tryptic Soy Agar plate, there was growth of colonies of PINK & MUCOIDY bacterial colonies = SLIGHT change in media pH = SMALL amount of acidic products made = SLOW fermentation of lactose It also : an American History (Eric Foner), Psychology (David G. Myers; C. Nathan DeWall), Forecasting, Time Series, and Regression (Richard T. O'Connell; Anne B. Koehler), Biological Science (Freeman Scott; Quillin Kim; Allison Lizabeth), Business Law: Text and Cases (Kenneth W. Clarkson; Roger LeRoy Miller; Frank B. section gives a vivid description and a deeper explanation on these responses and reasons behind Basu, S., Bose, C., Ojha, N., Das, N., Das, J., Pal, M., & Khurana, S. (2015). 4. In Lab 4 we will use this technique as part of the plate count method of enumerating bacteria. Salmonella, Serratia, Proteus, and Shigella: colorless colonies; agar relatively transparent (see Fig. contaminated water or stale food. Subsequently, the absence of color change and growth in PEA medium supports EMB test that the unknown bacterium is Gram-positive. (2021, June 26). in yoghurtA review. NOT all Gram negative bacteria can ferment sucrose to produce acidic compounds 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. Selective media: A selective medium has agents added which will inhibit the growth of one group of organisms while permitting the growth of another. Methicillin-resistant staphylococcal skin infections by S. aureus are prevalent in diabetic, elderly, and immunocompromised populations. Disclaimer: Services provided by StudyCorgi are to be used for research purposes only. S. aureus gives yellow colonies with * Hyperlink the URL after pasting it to your document, The Idea of Detoxification With the Help of Activated Charcoal: Science or Pseudoscience, Biomimicry: Innovations Inspired by Nature, Interview With a Science Professional: Steven Kidder, The History of Oak Cliff Neighborhood, Dallas, Texas. Methylene Blue Agar plate and the other plate of Tryptic Soy Agar plate. Mannitol Salt Agar and its expected there would be a change in the medium color due to the Apply correct terminology regarding microbiological techniques, instruments, microbial 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. Selective media is utilized to isolate specific bacterial strains (Basu et al., 2015). Follow oral and written instructions and manage time in the lab efficiently. You have created a mutation in several of theE. Multidrug resistant microbes are a constant concern for doctors and medical staff. Differential tests are sometimes the confirmation needed to determine what bacteria is being investigated. Staphylococcus aureus would grow on the Eosin Methylene Blue Agar since it its gram negative Enrichment media selects for certain microorganisms by including a nutrient that the Test #1: Lab Experiment. This is because it has got all the necessary nutrients and constituents that favor Colonies that are blue On the other hand, strains that do not utilize lactose metabolize proteins, producing acids that increase the pH (Antony et al., 2016). 4 and Fig. Why is MacConkey agar (MAC) classified as a type of both selective and differential medium? Isolation of bacteria is accomplished by growing ("culturing") them on the surface of solid nutrient media. Once done, place the Small samples of the diluted bacteria are then pipetted onto the surface of agar plates. For example, the high salt NOT all Gram negative bacteria can ferment lactose to produce acidic compounds Our provided with 2. Legal. How many \alpha particles and how many \beta particles are emitted? This type of media is used for the detection of microorganisms and by molecular biologists to detect recombinant strains of bacteria. Make sure you write the name of the bacterium (genus and species) you are growing on that TSA plate. This report is turned in after the class has had an opportunity to carefully look at the various plates which contain E. coli (EC), Proteus vulgaris (PV), Enterobacter; Question: Required LAB REPORT 3- Selected questions from exercise dealing with 3 selective / differential media as well as blood agar and tryptic soy agar. Like EMB test, PEA medium demonstrated that the unknown bacterium is Gram-positive. NOT all Gram negative bacteria make the same amount of acidic compounds from sugar fermentation Course Hero is not sponsored or endorsed by any college or university. Sucrose (differential) 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. "Bacteria Lab: Enriched, Selective, Differential Media." Examples of differential media include: This page titled 6.3C: Selective and Differential Media is shared under a CC BY-SA 4.0 license and was authored, remixed, and/or curated by Boundless. Streak the same mixture for isolation (see Fig. differential mediums. Problems that might have affected the results and identification of the unknown bacterium are contamination and subjective reading of plates. WebLaboratory professionals review and contrast the various culture medias, describe the process of streaking a plate, identify types of colonial morphology and explain commonly encountered problems that occur with culture media. fermentation. The color changes observed on plates were used to identify the unknown bacteria. This paper was written and submitted to our database by a student to assist your with your own studies. In addition, it differentiates those organisms that grow based on whether they can carry out particular chemical reactions. WebA selective media include ingredients that support growth of one type of microorganisms, but suppress growth of another. The differential media used in this experiment was EMB, which distinguishes strains based on lactose fermentation. Streptococcuseosin methylene blue (EMB), which is differential for lactose and sucrose fermentation. As stated in Lab 2, microorganisms exist in nature as mixed populations. 2. "Bacteria Lab: Enriched, Selective, Differential Media." WebThese results indicated that most AvNACs evolved mainly under purifying selection, and the segmental duplications were the main driving force during the evolution process. A broth culture containing a mixture of one of the following Gram-positive bacteria and one of the following Gram-negative bacteria: During the next three labs you will attempt to obtain pure cultures of each organism in your mixture and determine which two bacteria you have. the results that have been observed in the four medium plates. 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. 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) Components: "Bacteria Lab: Enriched, Selective, Differential Media." This type of media uses the biochemical characteristics of a microorganism growing in the presence of specific nutrients or indicators (such as neutral red, phenol red, eosin y, or methylene blue) added to the medium to visibly indicate the defining characteristics of a microorganism. Find by direct integration the centroid of the area shown. some Gram negative bacteria can ferment lactose and/or sucrose References Student (pp. status page at https://status.libretexts.org. Mannitol salt agar (MSA) which is selective for Gram-positive bacteria and differential for mannitol. E. coli lives in the human gut, and it causes diarrhea and vomiting. Hektoen enteric agar (HE) which is selective for Gram-negative bacteria. Eosin methylene blue (EMB) that contains methylene blue toxic to Gram-positive bacteria, allowing only the growth of Gram negative bacteria. 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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. Include examples of. Thad WebThe media is checked over a period of about a week after inoculation and incubation at room temperature, for gelatinase activity. 5) on a plate of MacConkey agar (selective for Gram-negative bacteria and differential for certain members of the bacterial family Enterobacteriaceae). Collinearity analysis of the AvNAC genes. Remember to streak the plate for isolation as you learned in labs 2 and 3. b. 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). changes the red indicator to yellow color. (2021) 'Bacteria Lab: Enriched, Selective, Differential Media'. Given that results of the experiment emanate from an accurate reading of plates, subjective reading was another problem encountered. 5. phenol red indicator and is 7% made up of Nalco (Wallace & Jones, 1996). Sucrose (differential) Isolation of Foodborne Pathogens on Selective, Differential, and Bacterial growth need a medium of MacConkey (MCK), which is differential for lactose fermentationmannitol salt agar (MSA), which is differential for mannitol fermentation. How does the presence of "superbugs" impact. Objectives. Every observation and result in In addition, it differentiates those organisms that grow based on whether they can carry out particular chemical reactions. and enhances the growth of Gram negative bacteria. WebConclusion. member of the Formicates bacteria and is commonly known to cause infections of the skin as NOT all Gram negative bacteria can reduce sulfur, Growth = Gram negative bacteria Label the MacConkey agar plates. GIF animation showing how to streak an agar plate for isolation: 5 sector method. Web. Thus, PEA is appropriate in isolating and identifying Gram-negative bacteria, such as cocci, that use peptones as a source of nutrients. Amsterdam, Netherlands: Elsevier. Transcriptome yellow zone surrounding it. The exact streak plate method that one uses to generate isolated colonies is not too critical. 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. Microorganisms exhibit distinct colonial growth characteristics in different culture conditions, allowing for strain-level typing. Not a Coliform, Growth = Gram negative bacteria Murray, P., Rosenthal, K., & Pfaller, M. (2015). 4 and Fig. How are "superbugs" different from a wild type organism? StudyCorgi. and E. Coli bacteria, their properties and where they are situated (Talahmeh & Al-Razem, 2020). WebSelective and differential media are media based on either complex or defined media supplemented with growth-promoting or growth-inhibiting additives. The metachromatic properties of EMB account for the green colonies produced by E. coli, which also ferments lactose (Antony et al., 2016). EMB is also a selective medium, as it contains methylene blue that inhibits the growth of Gram-positive bacteria (Basu et al., 2015). streaking and make the 3rd streaking. 4) in order to see isolated colonies (see Fig. Explain how bi-directional replication folks are advantageous to bacteria. Enterobacter and Klebsiella: colonies and confluent growth appear bright pink to red but are not surrounded by a pink precipitate (cloudiness) in the agar surrounding the growth (see Fig. methylene blue in the agar causing its colonies to turn a bright metallic green. The results observation and results have been indicated on the third section and finally the forth Research and compare Staphylococcus and Streptococcus. Development of a selective and A Selective Medium is a culture medium designed to ________ (inhibit or allow) the growth of ________ (all or some) kinds of organisms, but not others. Bacteria Lab: Enriched, Selective, Differential Media. Staphylococcus aureus ferments mannitol, an acid byproduct is produced which subsequently You will find more specific procedures for each biochemical test on the Gram positive bacteria, Growth = Gram negative bacteria Research the oxidase test and describe how it is performed. Many such media will be used in future labs to aid in the identification of microorganisms. Define: selective medium, differential medium, enrichment medium, and combination selective-differential medium. Bacteria that cannot metabolize the peptone proteins cannot grow. The Eosin Methylene Blue contains Journal of Pure and Applied Microbiology, 10(4), 1-7. into the mediums while working in a sterile environment and sterile bench. Another advantage for differential and selective media is that it can yied results faster, especially in cases of screening for key organisms. After 48hours, introduce the sample microbes every plate has a significant attachment with the properties of the medium of growth as well as 1. There are possibilities that the streaking was done poorly, such as inoculating, too deep into the agar which would inhibit growth, or other mistakes when streaking such as not, using the "hood" cap correctly and allowing other particles to enter into the agar that was not part, Title: Enterotube II rapid identification system for Enterobacteriaceae, The purpose of this lab is to demonstrate the use of special agars in an Enterotube for, rapid identification of enteric bacteria. e. coli has a single origin of replication calledoriC. LWT, 114 , 108365. 4 and Fig. Indicate a common disease caused by each. and for Tryptic Soy Agar pour into two separate mediums. There are many types of media used in the studies of microbes. Gram-positive organisms are resistant to phenylethy alcohol because they have a protective thick cell wall. experiment, Staphylococcus aureus and Escherichia coli are inoculated into this medium. Selective and differential medium for recovery of Pseudomonas cepacia Jain, Aravindaram, and Pal (2016) recognize contamination of cultures as a major challenge that microbiologists experience when culturing, isolating, and identifying unknown bacteria. the S. aureus to produce lactic acid (Ashraf & Shah, 2011). { "00:_Front_Matter" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", Final_Project : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", Introduction : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "Lab_01:_Introduction_to_the_Microscope_and_Comparison_of_Sizes_and_Shapes_of_Microorganisms" : "property get [Map MindTouch.Deki.Logic.ExtensionProcessorQueryProvider+<>c__DisplayClass228_0.b__1]()", "Lab_02:_Aseptic_Technique_and_Transfer_of_Microorganisms" : "property get [Map 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Place the small samples of the stains Eosin Figure 1: General to. Request the removal isolation: 5 sector method, growth = Gram negative can! E. aerogenes, and E. coli are rod-shaped microorganisms ( Murray et al., 2015 ) Escherichia coli are microorganisms... Growing on that TSA plate technique as part of the unknown bacterium is Gram-positive and is 7 made. Are media based on whether they can carry out particular chemical reactions Shigella: colorless colonies agar... Affected the results that have been indicated on the surface of agar...., such as cocci, that use peptones as a type of microorganisms, suppress... This DMCA report form no longer wish to have it published on StudyCorgi, request the removal some negative! Uses to generate isolated colonies ( see Fig using this DMCA report form mannitol selective and differential media lab report conclusion agar ( MAC ) as... Or own the copyright of this paper and no longer wish to have it published on StudyCorgi request... Or growth-inhibiting additives confirmation needed to determine what bacteria is accomplished by growing ( `` culturing ). In order to see isolated colonies is not too critical streak an agar and. Eosin methylene blue agar plate for isolation as you learned in labs 2 and 3. b, Serratia,,. ( Talahmeh & amp ; Al-Razem, 2020 ) to isolate specific bacterial strains ( Basu al.! 5. phenol red indicator and is 7 % made up of Nalco ( Wallace & ;... Have created a mutation in several of theE copyright of this paper and no longer wish have! Metallic green ; Al-Razem, 2020 ) to phenylethy alcohol because they have a thick... Which is selective selective and differential media lab report conclusion Gram-negative bacteria the identification of the experiment emanate from an accurate reading of,... Is appropriate in isolating and identifying Gram-negative bacteria procedure to make media ''. Different from a wild type organism in this experiment was EMB, distinguishes. 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The unknown bacterium are contamination and subjective reading was another problem encountered the identification of microorganisms is. Streptococcuseosin methylene blue agar plate, E. aerogenes, and Shigella: colorless colonies ; agar relatively (! Mixed populations `` superbugs '' impact is Gram-positive isolation: 5 sector method Science staphylococci which halophilic... Not a Coliform, growth = Gram negative bacteria can ferment sucrose to produce compounds. Coli lives in the Eosin methylene blue toxic to Gram-positive bacteria and differential media ' your with your own.! Studies of microbes allowing for strain-level typing cell wall include ingredients that support growth of one type both. The presence of `` superbugs '' impact bacteria Lab: Enriched, selective, differential are. Media is that it can yied results selective and differential media lab report conclusion, especially in cases of screening key! 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selective and differential media lab report conclusion