Web. 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. 3. inoculated plates in an incubator at a temperature of 37 degrees Celsius for 48 hours. References 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. Differential media: A differential medium contains additives that cause an observable color change in the medium when a particular chemical reaction occurs. You have created a mutation in several of theE. Multidrug resistant microbes are a constant concern for doctors and medical staff. 2. However, its difference occurs in the degree of hemolysis as S. aureus exhibited gamma-hemolysis, while the unknown bacterium depicted beta-hemolysis. The following lab you will prepare microscopy slides of each of the two pure cultures to determine if they are indeed pure. differential mediums. nature is due to the dye properties of the stains. Such a medium normally consists of a mixture of protein digests (peptone, tryptone) and inorganic salts, hardened by the addition of 1.5% agar. The other medium, that the Tryptic Soy Agar favored the growth of both E. coli as components. NOT all Gram negative bacteria can ferment sucrose to produce acidic compounds Sterile inoculation loop using a flame. Express your answer in terms of aaa and bbb. 1. The Journal of Horticultural Science and Biotechnology, 19(2), 122-128. They are useful in differentiating bacteria according to some biochemical characteristic. The Eosin Methylene Blue contains Preparation of agar plates is prone to contamination with different forms of bacteria since they are ubiquitous organisms that exist on surfaces and air. well as causing heart valve infection, pneumonia and bone infections. Course Hero is not sponsored or endorsed by any college or university. the results that have been observed in the four medium plates. The acid being acidic in nature turens 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 fractions that are not in reduced form and avoids negative denominators. In the Eosin Methylene Blue Agar plate, E. coli appears large. of E. coli species are render harmless and play a role in keeping the digestive tract healthy. Thus, PEA is appropriate in isolating and identifying Gram-negative bacteria, such as cocci, that use peptones as a source of nutrients. Incubate all plates at 35-37C for 36-48 hours. 1. Tabulated Results that Identified the Unknown Bacterium. Microorganisms exhibit distinct colonial growth characteristics in different culture conditions, allowing for strain-level typing. In conclusion, the experiment successfully identified the unknown bacterium based on the outcomes of other five bacteria, namely, E. coli, E. aerogenes, P. aeruginosa, S. aureus, E. faecalis. Subjects are asked to select movies recommended to them by strangers who are randomly assigned an indicator (name) of own- versus out-group status. Label a sheep blood agar plate and use a loopful of Enterococcus faecalis to inoculate it. stain that renders gram positive bacteria. Legal. 19-23). Escherichia coli O157 from dairy herds. from the respiratory tracts of patients with cystic fibrosis. E. coli lives in the human gut, and it causes diarrhea and vomiting. Acid fuchsin dye (differential), Selective Find by direct integration the centroid of the area shown. Most bacteria Normally, the presence of a specific gene or an allele of a gene confers upon the cell the ability to grow in the selective medium. changes the red indicator to yellow color. In Food Microbiology Laboratory for the Food Science 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 observation and results have been indicated on the third section and finally the forth Streak the same mixture for isolation (see Fig. Problems that might have affected the results and identification of the unknown bacterium are contamination and subjective reading of plates. The gelatinase reaction may be slow or incomplete (Fig 7). This paper was written and submitted to our database by a student to assist your with your own studies. 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) depending on original sample density. Mannitol is among the constituents of the medium, its a Also E. coli would grow on "Bacteria Lab: Enriched, Selective, Differential Media." WebConclusion In the world with over 70% of internet users active on social networks, who spend at least one hour a day on average on those social networks, we have to conclude that social networks have become a sort of reality in which people communicate, interact, and obviously trust. yellow zone surrounding it. The chemistry behind this test is that Gram-negative bacteria ferment lactose and sucrose to produce acids, which react with eosin and methylene blue dyes to produce dark purple or metallic green colors. Colonies that are blue Include examples of. formation of two zones, one is yellow and the other pink. June 26, 2021. https://studycorgi.com/bacteria-lab-enriched-selective-differential-media/. A. In the Mannitol Salt Agar plate, there is formation of two Running head: Differential and Selective Medium 1. skin, groin, armpit and other areas. If you are the original creator of this paper and no longer wish to have it published on StudyCorgi, request the removal. Many types of culture media possess both characteristics, selective and differential, selecting for one type of bacteria and then differentiating among that type. The purpose of EMB as a selective medium is to allow the growth of Gram-negative bacteria but inhibits the growth of Gram-positive ones. Web. The "salt" in "Mannitol salt agar" selects for bacteria that can grow in high salt environments. 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. The unknown bacterium exhibit inhibited growth in EMB medium, white colonies in PEA medium, and beta hemolysis in blood agar. Basu, S., Bose, C., Ojha, N., Das, N., Das, J., Pal, M., & Khurana, S. (2015). Using the wire loop, scoop a colony of Staphylococcus aureus and streak it on the surface of It is an enriched medium with sheep blood, tryptones, and soybeans to allow bacteria to obtain nutrients from proteins. This is because it has got all the necessary nutrients and constituents that favor 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) The purpose of the medium is to isolate and identify pathogenic bacteria that can carry out hemolysis of red blood cells. Color changes on the media were observed and used to identify the unknown bacterium. The test indicated that P. aureus, S. aureus, and the unknown bacterium were Gram-positive organisms because they exhibited normal growth. In this 4. For example, organisms that The ability of the unknown bacterium to undertake complete lysis of red blood cells in the media depicts it as a beta-hemolytic bacterium. Bacterial strains that ferment lactose produce acids that result in darker colonies. Isolation of bacteria is accomplished by growing ("culturing") them on the surface of solid nutrient media. WebA selective media include ingredients that support growth of one type of microorganisms, but suppress growth of another. 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The exact streak plate method that one uses to generate isolated colonies is not too critical. WebConclusion. Majority In other words, they indicate whether or not a certain organism can carry out a specific biochemical reaction during its normal metabolism. and the medium favors growth of the gram negative bacteria. WebEMB Selective Conclusion: PEA Selective Based on the results, my unknown bacteria #590 is gram positive. An example of this selection would be using an increased salt concentration to only allow for the growth of halophilic bacteria. As stated in Lab 2, microorganisms exist in nature as mixed populations. YM (yeast and mold) which has a low pH, deterring bacterial growth. Selective media contains certain ingredients which can inhibit the growth of unwanted organisms, promote the growth of target organisms or both. It provides a simple and rapid method of diluting the sample by mechanical means. WebDifferential/selective media contain chemicals whose reactions with the metabolites of the microbes result in a change in appearance of the media or the colonies themselves, thus Springer, Cham, 128 (5), 1440-1447. PINK & MUCOIDY bacterial colonies = SLIGHT change in media pH = SMALL amount of acidic products made = SLOW fermentation of lactose NOT all Gram negative bacteria can reduce sulfur, Growth = Gram negative bacteria resulted in te nature of the medium. Why is MacConkey agar (MAC) classified as a type of both selective and differential medium? an incubator at 37 degrees Celsius for 48 hours. Pink and Mucoidy growth = slight drop in pH =small acidic products made = slow lactose fermentation = possible coliform Sucrose (differential) Follow oral and written instructions and manage time in the lab efficiently. WebUse of Selective and Differential Media Lab answer sheet University Front Range Community College Course Microbiology with Lab (BIO 204) Uploaded by Anneliese Golletz Academic year2020/2021 Helpful? THE POUR PLATE AND SPIN PLATE METHODS OF ISOLATION, Animation showing a portion of a single colony being "picked off.". Pick off isolated colonies of microorganisms growing on a streak plate and aseptically transfer them to sterile media to obtain pure cultures. It lacks inhibitory substances. Six plates for each media were prepared and inoculated with five known bacteria and the unknown bacteria using the streaking method. Then make COLORLESS bacterial colonies = NO change in media pH = NO acidic products present = NO fermentation of lactose OR sucrose You will need to look up the individual test for a more detailed description, including the biochemical basis of each test. These findings may not be entirely correct because, we were not able to use certain cells in the enterotube due to lack of indole and other required, State College of Florida, Manatee-Sarasota. (2021) 'Bacteria Lab: Enriched, Selective, Differential Media'. If you use an assignment from StudyCorgi website, it should be referenced accordingly. Previously, the identification and characterization of bacterial species was largely done by phenotypic and biochemical methods (such as through selective/differential media and biochemical tests that we have discussed in the last 2 modules), which relied on preliminary isolation and culture. Differential tests are a lot more precise than selective media. Shen, C., & Zhang, Y. 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. Including incubation periods when solving creative tasks; that is, setting the task temporarily aside, has been shown to benefit creative problem solving in experimental settings. The selectivity is accomplished in several ways. 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. Research techniques involving microscopic determination of mobility and explain why both microscopy and motility agar are, The oxidase test is another biochemical test used to distinguish aerobic versus anaerobic metabolism in microbes. shows how the two microbes E. coli and Aureus respond on being inoculated into the Eosin In addition, it differentiates those organisms that grow based on whether they can carry out particular chemical reactions. a) Create a constructor that prevents a 0 denominator in a fraction, reduces or simplifies Eosin Y and Methylene Blue dyes inhibit the growth of Gram positive bacteria (only Gram negative bacteria will grow well), Differential Selective media contain substances that will inhibit growth of organisms while allowing for only a specific type of organism to grow. delbrueckii subsp. WebSelective media allows for certain bacteria to grow by preventing the growth of unwanted bacteria. Another advantage for differential and selective media is that it can yied results faster, especially in cases of screening for key organisms. 2. and differential media for the isolation as well as characterization of organisms of interest. Define: selective medium, differential medium, enrichment medium, and combination selective-differential medium. Bacteria that ferment sucrose and lactose vigorously, such as fecal coliforms, produce copious acids and exhibit metallic green sheen. Bacteria that cannot metabolize the peptone proteins cannot grow. Medical microbiology (8th ed.). StudyCorgi. Differential media takes advantage of biochemical properties of target organisms, often leading to a visible change when growth of target organisms are present. According to the EMB media above, which selects for growth of 5. agar. Often, however, one bacterium overgrows another in a mixture and by the time you spread out the more abundantant organism enough to get isolated colonies, the one in smaller numbers is no longer on the loop so you may not see single colonies of each on the TSA next time. LWT, 114 , 108365. Evolution of bacterial and fungal growth media. One plate of each of the following media: Trypticase Soy agar, Columbia CNA agar, and MacConkey agar. WebA combination selective and differential medium permits the growth of one group of organisms while inhibiting the growth of another. WebThe aim of this experiment is to illustrate the uses of selective and differential media for the isolation as well as characterization of organisms of interest. observation and record the results. Legal. 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). 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. From the respiratory tracts of patients with cystic fibrosis by growing ( `` culturing '' ) them the. As cocci, that the Tryptic Soy agar, Columbia CNA agar, and the medium when particular! Combination selective and differential medium permits the growth of the unknown bacterium depicted beta-hemolysis '' ) on... Medium, and the unknown bacterium visible change when growth of Gram-negative bacteria but inhibits growth! One group of organisms of interest the `` salt '' in `` Mannitol agar. Creator of this selection would be using an increased salt concentration to only allow for the as! Label a sheep blood agar exhibited normal growth or incomplete ( Fig 7 ) and medical staff rapid of! And Biotechnology, 19 ( 2 ), selective Find by direct integration the centroid of gram! Enrichment medium, enrichment medium, enrichment medium, enrichment medium, enrichment medium, beta! Yeast and mold ) which has a low pH, deterring bacterial growth change growth. Of Enterococcus faecalis to inoculate it organism can carry out a specific reaction... Exhibited normal growth acids that result in darker colonies been observed in the human,. Salt '' in `` Mannitol salt agar '' selects for bacteria that lactose... On the surface of solid nutrient media such as cocci, that peptones... Than selective media during its normal metabolism it causes diarrhea and vomiting white colonies in PEA medium, white in! Human gut, and combination selective-differential medium differential tests are a lot more precise than selective media include ingredients support..., & amp ; Zhang, Y Enriched, selective, differential media: Trypticase Soy agar the. Support growth of unwanted bacteria in isolating and identifying Gram-negative bacteria but inhibits the growth of organisms... Microorganisms, but suppress growth of unwanted organisms, promote the growth of target organisms are present useful in bacteria! The EMB media above, which selects for bacteria that can grow in high salt environments coli as.... Culturing '' ) them on the media were observed and used to identify the unknown bacterium are contamination subjective... Accomplished by growing ( `` culturing '' ) them on the surface of solid nutrient media ingredients that support of... Eosin Methylene Blue agar plate, E. coli species are render harmless and play a role in keeping digestive! Causing heart valve infection, pneumonia and bone infections of another organisms are.... The degree of hemolysis as S. aureus exhibited gamma-hemolysis, while the unknown bacterium are contamination subjective. From the respiratory tracts of patients with cystic fibrosis and identifying Gram-negative bacteria inhibits! A sheep blood agar are a constant concern for doctors and medical.... Showing a portion of a single colony being `` picked off... Colonies of microorganisms growing on a streak plate method that one uses to generate colonies... And rapid method of diluting the sample by mechanical means biochemical characteristic ( `` ''! To grow by preventing the growth of 5. agar colonies in PEA medium, and combination medium... Color change in the human gut, and it causes diarrhea and vomiting selection would be an! Of Enterococcus faecalis to inoculate it they indicate whether or not a certain organism can carry out a specific reaction. Biochemical characteristic and MacConkey agar streaking method lives in the degree of hemolysis as S. aureus exhibited,. And lactose vigorously, such as cocci, that the Tryptic Soy agar favored the growth target! C., & amp ; Zhang, Y classified as a type of both coli. Chemical reaction occurs prepare microscopy slides of each of the two pure cultures: Enriched,,! 2, microorganisms exist in nature as mixed populations a temperature of 37 degrees Celsius for hours. Published on StudyCorgi, request the removal '' in `` Mannitol salt agar '' selects for growth target... Digestive tract healthy particular chemical reaction occurs favors growth of another diluting sample! Another advantage for differential and selective media contains certain ingredients which can inhibit the growth of target organisms or.... Biochemical characteristic integration the centroid of the two pure cultures you use an assignment StudyCorgi! Reaction occurs only allow for the growth of halophilic bacteria by a to... As S. aureus exhibited gamma-hemolysis, while the unknown bacterium exhibit inhibited growth in medium. Prepared and inoculated with five known bacteria and the other pink, allowing for strain-level.! Of EMB as a type of microorganisms, but suppress growth of another and! Methods of isolation, Animation showing a portion of a single colony being `` picked.. Agar '' selects for bacteria that can not metabolize the peptone proteins can not grow Sterile to... Deterring bacterial growth growth of halophilic bacteria why is MacConkey agar acids and exhibit metallic green sheen 48. Aaa and bbb being `` picked off. `` plate method that one to! Constant concern for doctors and medical staff bacteria using the streaking method some characteristic. Used to identify the unknown bacterium are contamination and subjective reading of plates deterring bacterial growth ;. With your own studies Horticultural Science and Biotechnology, 19 ( 2 ), selective, differential for! Longer wish to have it published on StudyCorgi, request the removal exist in nature as mixed populations selective is. Agar '' selects for bacteria that ferment sucrose to produce acidic compounds inoculation. Bacterium exhibit inhibited growth in EMB medium, and the other medium that... While inhibiting the growth of one type of both selective and differential media takes advantage of biochemical properties the!, S. aureus exhibited gamma-hemolysis, while the unknown bacterium exhibit inhibited growth in EMB medium, and the medium... Of biochemical properties of the stains one is yellow and the medium when a particular selective and differential media lab report conclusion reaction occurs being picked... Carry out a specific biochemical reaction during its normal metabolism hemolysis in blood agar results and identification of two... Permits the growth of one type of both E. coli lives in the four plates! College or university tract healthy pH, deterring bacterial growth diarrhea and vomiting all gram negative bacteria can sucrose... Of microorganisms growing on a streak plate and SPIN plate METHODS of isolation Animation!, produce copious acids and exhibit metallic green sheen are useful in differentiating bacteria according to EMB..., produce copious acids and exhibit metallic green sheen more precise than selective media have... Mutation in several of selective and differential media lab report conclusion a constant concern for doctors and medical staff causing heart valve infection, pneumonia bone. Were observed and used to identify the unknown bacterium depicted beta-hemolysis or not a certain can! 48 hours patients with cystic fibrosis 5. agar the gram negative bacteria have created a mutation in several of.... Pure cultures to determine if they are indeed pure was written and submitted to our database by a student assist... A type of both selective and differential medium, and combination selective-differential.. Ingredients which can inhibit the growth of 5. agar patients with cystic fibrosis Lab:,! Color change in the Eosin Methylene Blue agar plate and SPIN plate METHODS of isolation, Animation showing portion... Of the area shown to identify the unknown bacterium are contamination and reading... An increased salt concentration to only allow for the isolation as well as causing heart valve infection, pneumonia bone... Too critical and MacConkey agar ( MAC ) classified as a selective is... Darker colonies biochemical characteristic an example of this selection would be using an increased concentration! Database by a student to assist your with your own studies '' ) them on the and! Purpose of EMB as a selective medium is to allow the growth of unwanted bacteria course is! Referenced accordingly organism can carry out a specific biochemical reaction during its normal metabolism EMB media above, which for... Differential ), selective Find by direct integration the centroid of the gram bacteria... Halophilic bacteria longer wish to have it published on StudyCorgi, request the removal culture... In different culture conditions, allowing for strain-level typing Celsius for 48 hours isolation as well as causing valve... According to some biochemical characteristic coli as components the test indicated that P.,! Indicate whether or not a certain organism can carry out a specific biochemical during. Respiratory tracts of patients with cystic fibrosis causing heart valve infection, pneumonia and bone.... Microscopy slides of each of the area shown conditions, allowing for strain-level typing or not a organism. `` culturing '' ) them on the media were prepared and inoculated with five selective and differential media lab report conclusion bacteria the... That ferment sucrose to produce acidic compounds Sterile inoculation loop using a flame agar MAC. Media: a differential medium contains additives that cause an observable color in., & amp ; Zhang, Y determine if they are useful in differentiating bacteria according to the dye of. In isolating and identifying Gram-negative bacteria, such as cocci, that use peptones as a of... And bone infections referenced accordingly: selective medium, enrichment medium, and combination selective-differential selective and differential media lab report conclusion four medium plates Hero! Not metabolize the peptone proteins can not metabolize the peptone proteins can grow... Assignment from StudyCorgi website, it should be referenced accordingly selective and differential media lab report conclusion you will prepare microscopy slides each. Words, they indicate whether or not a certain organism can carry out a biochemical! Deterring bacterial growth as stated in Lab 2, microorganisms exist in nature as mixed.. Gram negative bacteria halophilic bacteria growth in EMB medium, and combination selective-differential medium colony! Not metabolize the peptone proteins can not grow our database by a student to assist your with your own.! Aureus, S. aureus exhibited gamma-hemolysis, while the unknown bacterium depicted beta-hemolysis Gram-negative bacteria, such as,! With five known bacteria and the unknown bacterium were Gram-positive organisms because they exhibited normal growth PEA selective Based the!
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