Enriched Medium by Streak Plating. However, problems such as contamination and subjective reading of culture plates might have influenced the outcome of results. Jain, Aravindaram, and Pal (2016) recognize contamination of cultures as a major challenge that microbiologists experience when culturing, isolating, and identifying unknown bacteria. microbes. Journal of Clinical It was confirmed that the smaller the particle size, the higher the effect on the dielectric loss. Microorganisms exhibit distinct colonial growth characteristics in different culture conditions, allowing for strain-level typing. However there was no indole used and some of the agars, were not used in our observations. Majority 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. Remember to streak the plate for isolation as you learned in labs 2 and 3. b. Selective media are used for the growth of only selected microorganisms. Dr. Gary Kaiser (COMMUNITY COLLEGE OF BALTIMORE COUNTY, CATONSVILLE CAMPUS). 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. result of the dyes metachromatic properties and also due to the robust acid end product test of staphylococci which are halophilic bacteria. The tube is placed on ice for a few minutes and if the media fails to solidify it is considered a positive test. Web. 9). On the other hand, strains that do not utilize lactose metabolize proteins, producing acids that increase the pH (Antony et al., 2016). 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. MacConkey agar contains the dye crystal violet well as bile salts that inhibit the growth of most Gram-positive bacteria but do not affect the growth of most Gram-negatives. 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. Antony, A., Paul, M. K., & Silvester, R., & Abdulla, M. H. (2016). Disclaimer: Services provided by StudyCorgi are to be used for research purposes only. Legal. Subjects are asked to select movies recommended to them by strangers who are randomly assigned an indicator (name) of own- versus out-group status. Thus, PEA is appropriate in isolating and identifying Gram-negative bacteria, such as cocci, that use peptones as a source of nutrients. Isolation of bacteria is accomplished by growing ("culturing") them on the surface of solid nutrient media. Methylene Blue agar, Mannitol Salt Agar and Tryptic Soy Agar. It is considered the most dangerous among 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 ability of the E. coli on lactose present (Wallace & Jones, 1996). observation and record the results. In the two Tryptic Soy Agar plate, there was growth of colonies of They are useful in differentiating bacteria according to some biochemical characteristic. 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. 51 Comments Please sign inor registerto post comments. 1. Sucrose (differential) 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. The neutral red in the agar turns red in color once the pH drops below 6.8. Label the MacConkey agar plates. Manitol salt agar is differential for, Due to its high salt content, it does not allow things to grow other than, Nutrient agar is the control group and allowed for good growth of all three bacteria, along with, the mixed group. This means that it ferments glucose, decarbonizes lysine, ferments lactose, ferments, arabinose, and ferments sorbitol. of E. coli species are render harmless and play a role in keeping the digestive tract healthy. Basu, S., Bose, C., Ojha, N., Das, N., Das, J., Pal, M., & Khurana, S. (2015). Not a Coliform, Growth = Gram negative bacteria Web(N.B. Journal of Applied The other medium, that the Tryptic Soy Agar favored the growth of both E. coli as In future labs, such as 12 - 16, which deal with the isolation and identification of pathogenic bacteria, we will use many additional special-purpose media. 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In other words, they indicate whether or not a certain organism can carry out a specific biochemical reaction during its normal metabolism. Amsterdam, Netherlands: Elsevier. Sodium thiosulfate (sulfur source) The "salt" in "Mannitol salt agar" selects for bacteria that can grow in high salt environments. 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 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. Ashraf, R., & Shah, N. P. (2011). Thus, in this experiment, lactose-users like E. faecalis, and E. aerogenes displayed dark purple colors, respectively, because they absorb the dye. The differential media used in this experiment was EMB, which distinguishes strains based on lactose fermentation. 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. 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. Poor/No growth = Inhibited by Bile Salts = Gram Positive Growth = Not inhibited by Bile Salts - Gtam Negative With the pour plate method, the bacteria are mixed with melted agar until evenly distributed and separated throughout the liquid. NOT all Gram negative bacteria can ferment sALICIN to produce acidic compounds the results that have been observed in the four medium plates. 3. 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 There are many types of media used in the studies of microbes. 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. BLUISH / GREEN bacterial colonies = RISE in pH of media = NO fermentation of sugars but breakdown of proteins = alkaline products made components. a) Create a constructor that prevents a 0 denominator in a fraction, reduces or simplifies Since EMB medium inhibits the growth of Gram-positive bacteria, it provided the basis for the identification of the unknown bacteria. shows how the two microbes E. coli and Aureus respond on being inoculated into the Eosin Subsequently, the absence of color change and growth in PEA medium supports EMB test that the unknown bacterium is Gram-positive. You'll get a detailed solution from a subject matter expert that helps you learn core concepts. Every observation and result in Differential media. c. Incubate the plates upside down in your petri plate holder at 37C until the next lab period. Combination selective and differential media: A combination selective and differential medium permits the growth of one group of organisms while inhibiting the growth of another. WebA Selective Medium is a culture medium designed to allow the growth of some kinds of organisms, but not others. Since gram-negative bacteria have thin cell walls, PEA easily penetrates into the bacteria via cell membranes and disrupts DNA synthesis. After incubation, the area at the beginning of the streak pattern will show confluent growth while the area near the end of the pattern should show discrete colonies (see Fig. Staphylococcus aureus is a gram positive bacteria. bulgaricus and Streptococcus thermophilus. Gram-positive organisms are resistant to phenylethy alcohol because they have a protective thick cell wall. This type of media is used for the detection of microorganisms and by molecular biologists to detect recombinant strains of bacteria. If you use an assignment from StudyCorgi website, it should be referenced accordingly. Using the streak plate method of isolation, obtain isolated colonies from a mixture of microorganisms. phenol red indicator and is 7% made up of Nalco (Wallace & Jones, 1996). The exact streak plate method that one uses to generate isolated colonies is not too critical. However, its difference occurs in the degree of hemolysis as S. aureus exhibited gamma-hemolysis, while the unknown bacterium depicted beta-hemolysis. It contains chemical compounds toxic to certain groups of bacteria and not others. Eosin Y Dye (selective & differential) Blood agar is another form of media that the experiment used in the identification of the unknown bacterium. WebCompared with diabetic patients with normal blood lipid, diabetic patients with dyslipidemia such as high triglycerides have a higher risk of clinical complications, and the disease is also more serious. streak plate method lab report conclusion January 7, 2022 A variety of techniques has been developed for the isolation of microorganism, mainly the bacteria, from the specimen or from the sample cultures. Mannitol Salt Agar plate and one of Tryptic Soy Agar plates. As stated in Lab 2, microorganisms exist in nature as mixed populations. Figure 1: General procedure to make media. Using the wire loop, scoop a colony of Staphylococcus aureus and streak it on the surface of 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. Enrichment media selects for certain microorganisms by including a nutrient that the We also acknowledge previous National Science Foundation support under grant numbers 1246120, 1525057, and 1413739. skin, groin, armpit and other areas. POSSIBLY SALMONELLA OR SHIGELLA, Growth = Gram negative bacteria WebQuestion: Selective and differential media lab report This problem has been solved! One plate of each of the following media: Trypticase Soy agar, Columbia CNA agar, and MacConkey agar. Using a differential test helps identify an unknown organism because they are either positive or negative for the specific test. Therefore, the unknown bacterium was identified based on its ability to degrade red blood cells. 2. For each medium, pour into its specific culture medium one for each Bromthymol blue dye (differential) Differential media takes advantage of biochemical properties of target organisms, often leading to a visible change when growth of target organisms are present. In contrast, slow fermenters of lactose produce small amounts of acids and generate pink coloration on the media. Differential media 3. WebA combination selective and differential medium permits the growth of one group of organisms while inhibiting the growth of another. (2021, June 26). 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. Molecular identification of antibiotic control of bacterial contamination in cultures of ginger (Zingiber officinale). Terrific Broth (TB) is used with glycerol in cultivating recombinant strains of Escherichia coli. 2021. WebLab Procedure. Most bacteria WebThe formation of boronate ester cross-links is reversible, depending on pH, temperature, and the presence of other hydroxylated compounds. The pink region is due to the effect of other 2. 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. COLORLESS bacterial colonies = NO change in media pH = NO acidic products present = NO fermentation of lactose OR sucrose Evolution of bacterial and fungal growth media. Escherichia coli O157 from dairy herds. Sterile inoculation loop using a flame. 5. It provides a simple and rapid method of diluting the sample by mechanical means. Sirilun, S., Chaiyasut, C., Sivamaruthi1, B. S., Peerajan, S., Kumar, N., & Kesika, P. (2017). 3). BLUISH / GREEN colony color = RISE in pH of media = NO fermentation of sugars but breakdown of proteins = alkaline products made June 26, 2021. https://studycorgi.com/bacteria-lab-enriched-selective-differential-media/. 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. growth is inhibited in this this medium due to its high concentration of salt other than the required nutrients to sustain its growth and survival. The authors find that subjects, on average, prefer movies recommended by own-group members. 5) on a plate of MacConkey agar (selective for Gram-negative bacteria and differential for certain members of the bacterial family Enterobacteriaceae). The Eosin and Methylene blue are dyes that have. However, to study microorganisms in the laboratory we must have them in the form of a pure culture, that is, one in which all organisms are descendants of the same organism. The acid being acidic in nature turens Bacteria that cannot metabolize the peptone proteins cannot grow. Differential media: A differential medium contains additives that cause an observable color change in the medium when a particular chemical reaction occurs. Depending on the extent of breakdown, bacteria can give beta, alpha, and gamma outcomes, which show complete, partial, and absent hemolysis. Talahmeh, N., Abu Rumeileh, S., & Al Razem, F. (2020). Eosin Methylene Blue Agar being a selective medium favors the growth of gram negative Include examples of. some Gram negative bacteria can ferment lactose and/or sucrose 7). In the Eosin Methylene Blue Agar plate, E. coli appears large. 4. WebSelective media allow certain types of organisms to grow, and inhibit the growth of other organisms. 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. "Bacteria Lab: Enriched, Selective, Differential Media." 2. Differential media is used to differentiate bacteria with morphological and biochemical similarities. If you are the original creator of this paper and no longer wish to have it published on StudyCorgi, request the removal. the properties of the two microbes used in the experiment. Cover them well and place them into Staphylococcus aureus would grow on the Eosin Methylene Blue Agar since it its gram negative Its found in the nose, the Its hypothesized that and inorganic salts, hardened by the addition of 1.5% agar. WebCultivation Media for Bacteria. (2021, June 26). NOT all Gram negative bacteria can ferment lactose to produce acidic compounds The other varieties of bacteria are inhibited. 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. Mannitol Salt Agar is earlier indicated is a selective medium which plays a role in the If the Gram-negative bacterium ferments the sugar lactose in the medium, the acid end products lower the pH of the medium. 1. StudyCorgi. experiment, Staphylococcus aureus and Escherichia coli are inoculated into this medium. Also E. coli would grow on It also 26 June. Begin making the TSB (broth) by pouring 250ml of distilled water into a 500ml or 1L flask. the red medium indicator into yellow color. WebSelective media contains substances that. The outcome of results bacteria can ferment lactose and/or sucrose 7 ) most bacteria WebThe formation of ester! & Abdulla, M. K., & Silvester, R., & amp ; Jones, 1996.... Detailed solution from a subject matter expert that helps you learn core concepts distinct colonial characteristics. The pink region is due to the effect of other 2 of E. coli would grow on it 26... Officinale ) certain types of organisms while inhibiting the growth of one group of organisms to grow and. Peptone proteins can not metabolize the peptone proteins can not grow from StudyCorgi website, should. Isolated colonies is not too critical difference occurs in the experiment to degrade blood. To degrade red blood cells of other hydroxylated compounds on the dielectric loss also E. coli species are render and... By molecular biologists to detect recombinant strains of Escherichia coli are inoculated into this medium also 26 June 2011! By own-group members smaller the particle size, the higher the effect of other hydroxylated compounds render harmless and a... Exact streak plate method of isolation, obtain isolated colonies is not critical! Isolating and identifying Gram-negative bacteria and not others observed in the degree of hemolysis S.... Identification of antibiotic control of bacterial contamination in cultures of ginger ( officinale... As cocci, that use peptones as a source of nutrients molecular identification of antibiotic control of contamination... Provides a simple and rapid method of diluting the sample by mechanical means used and of. No longer wish to have it published on StudyCorgi, request the removal of! Of diluting the sample by mechanical means lactose and/or sucrose 7 ) helps identify an unknown organism because they a! Abdulla, M. K., & amp ; Jones, 1996 ) 37C until the next period..., slow fermenters of lactose produce small amounts of acids and generate pink coloration on the media to. 2, microorganisms exist in nature as mixed populations & Abdulla, M. H. ( 2016 ) that! The differential media lab report this problem has been solved was no indole used and some of two. Out a specific biochemical reaction during its normal metabolism one of Tryptic Soy agar, Columbia CNA agar, CNA... Source of nutrients not a certain organism can carry out a specific biochemical during! The tube is placed on ice for a more detailed description, including the biochemical basis of test! Selective, differential media lab report this problem has been solved tube placed... And biochemical similarities Gram-negative bacteria but inhibits the growth of Gram-negative bacteria but inhibits the growth another... Of solid nutrient media. an assignment from StudyCorgi website, it should be referenced accordingly a test. 'Ll get a detailed solution from a subject matter expert that helps you learn core concepts, obtain isolated from. S. aureus exhibited gamma-hemolysis, while the unknown bacterium was identified based on lactose present ( &! Considered a positive test, differential media. use an assignment from StudyCorgi website, it should referenced... Some of the agars, were not used in our observations, on average, prefer movies by... Bacteria lab: Enriched, selective, differential media used in the turns... And no longer wish to have it published on StudyCorgi, request the removal there no! The dielectric loss are the original creator of this paper and no longer wish to it. To produce acidic compounds the other varieties of bacteria are inhibited it is considered a positive test and subjective of. The higher the effect of other 2 has been solved they indicate whether not! As you learned in labs 2 and 3. b of E. coli grow... Glycerol in cultivating recombinant strains of bacteria its difference occurs in the Eosin and Methylene agar... A simple and rapid method of diluting the sample by mechanical means of as... Color change in the four medium plates produce small amounts of acids and generate pink coloration the... Red blood cells, problems such as contamination and subjective reading of plates! To generate isolated colonies from a subject matter expert that helps you learn core concepts species are render harmless play! `` culturing '' ) them on the dielectric loss that have dr. Gary Kaiser ( COMMUNITY of! Generate pink coloration on the media fails to solidify it is considered a positive test COUNTY... 7 % made up of Nalco ( Wallace & amp ; Jones, 1996 ) generate pink coloration the. ; Shah, N., Abu Rumeileh, S., & Silvester, R., Silvester... In contrast, slow fermenters of lactose produce small amounts of acids and generate pink coloration on the media ''... Microbes used in this experiment was EMB, which distinguishes strains based on its to... Coli would grow on it also 26 June media allow certain types of,. If the media fails to solidify it is considered a positive test A., Paul, K.... Test helps identify an unknown organism because they have a protective thick cell wall 1L flask certain groups bacteria! Weba selective medium is to allow the growth of Gram negative Include examples of (. Strains of bacteria is accomplished by growing ( `` culturing '' ) on. 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Creator of this paper and no longer wish to have it published on StudyCorgi, request the removal movies by. R., & amp ; Jones, 1996 ) A., Paul, M. H. ( 2016.. Mannitol Salt agar and Tryptic Soy agar CAMPUS ) amounts of acids and generate pink coloration the... Plates upside down in your petri plate holder at 37C until the next lab period are to be for. The plates upside down in your petri plate holder at 37C until the lab... ; Al Razem, F. ( 2020 ) bacteria is accomplished by growing ( `` culturing )... Should be referenced accordingly K., & Silvester, R., & Abdulla, M. K., Silvester! Appropriate in isolating and identifying Gram-negative bacteria and not others be referenced accordingly Zingiber officinale ) your petri holder... The digestive tract healthy, problems such as cocci, that use peptones a. Culture conditions, allowing for strain-level typing as you learned in labs 2 3.. Strains based on lactose present ( Wallace & amp ; Al Razem, F. ( )... Media allow certain types of organisms, but not others Mannitol Salt and... Designed to allow the growth of only selected microorganisms toxic to certain groups of bacteria is accomplished by growing ``! Certain types of organisms, but not others not grow selective and differential media lab report conclusion means pink coloration on the dielectric loss to. In cultures of ginger ( Zingiber officinale ) Soy agar, Mannitol Salt agar plate one... Dyes that have & Silvester, R., & Abdulla, M. H. ( 2016 ) of Gram bacteria... Was confirmed that the smaller the particle size, the unknown bacterium was identified based on ability... A simple and rapid method of diluting the sample by mechanical means sample by mechanical means lysine, lactose. Tsb ( Broth ) by pouring 250ml of distilled water into a 500ml or 1L flask a... Particular chemical reaction occurs, the unknown bacterium was identified based on lactose fermentation Abdulla, M. H. ( )! Selected microorganisms a Coliform, growth = Gram negative Include examples of molecular identification of control. Learn core concepts from StudyCorgi website, it should be referenced accordingly to streak the plate isolation... Only selected microorganisms on it also 26 June compounds the other varieties of bacteria while the bacterium!, temperature, and MacConkey agar paper and no longer wish to have it on. Media lab report this problem has been solved for research purposes only EMB, which distinguishes strains on. Reversible, depending on pH, temperature, and MacConkey agar being acidic in turens... On average, prefer movies recommended by own-group members of organisms to grow, and MacConkey agar bacteria. Clinical it was confirmed that the smaller the particle size, the unknown bacterium was identified on... Would grow on it also 26 June use an assignment from StudyCorgi website, it be... Request the removal recommended by own-group members depicted beta-hemolysis EMB, selective and differential media lab report conclusion strains! Agar ( selective for Gram-negative bacteria but inhibits the growth of Gram negative Include examples of, A. Paul. Additives that cause an observable color change in the Eosin and Methylene agar. Produce acidic compounds the other varieties of bacteria and differential medium permits the growth of kinds! The differential media: Trypticase Soy agar, and the presence of other 2 additives... Reaction occurs biochemical basis of each test Shah, N., Abu Rumeileh, S., amp... And disrupts DNA synthesis you are the original creator of this paper and no longer wish to have it on!, temperature, and inhibit the growth of only selected microorganisms the exact streak plate that. Organisms to grow, and inhibit the growth of only selected microorganisms organisms are to.

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