Running head: Differential and Selective Medium 1. the S. aureus to produce lactic acid (Ashraf & Shah, 2011). 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Web. Incubate the three plates upside down and stacked in the petri plate holder on the shelf of the 37C incubator corresponding to your lab section until the next lab period. StudyCorgi. 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. Incubate all plates at 35-37C for 36-48 hours. The differential media used in this experiment was EMB, which distinguishes strains based on lactose fermentation. Include examples of. Bacteria that cannot metabolize the peptone proteins cannot grow. Single colonies will be visible on the 2nd or the 3rd streak. X-gal plates, which are differential for lac operon mutants. 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. CompareandContrast\text{\red{Compare and Contrast}}CompareandContrast Describe the difference between a reaction that often happens on its own and one that does not. Normally, pathogenic bacteria belonging to genus Streptococcus have endotoxins that they use in breaking down red blood cells and deriving nutrients. 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. This means that it ferments glucose, decarbonizes lysine, ferments lactose, ferments, arabinose, and ferments sorbitol. The introduction 2. coliform bacterium that is mostly found in the intestines and in the gut of some animals. The Eosin Methylene Blue contains Orange/Yellow Growth = drop on pH = Large acidic = products made = fermentation of sugars = Not Salmonella or Shigella MacConkey (MCK), which is differential for lactose fermentationmannitol salt agar (MSA), which is differential for mannitol fermentation. Possible Coliform, Growth = Gram negative bacteria Like EMB test, PEA medium demonstrated that the unknown bacterium is Gram-positive. Microbial characteristics of culture media are used to identify and characterize bacteria. Ashraf, R., & Shah, N. P. (2011). Sucrose (differential) Express your answer in terms of aaa and bbb. and enhances the growth of Gram negative bacteria. Transcriptome Differential media This type of media is used for the detection of microorganisms and by molecular biologists to detect recombinant strains of bacteria. After 48hours, introduce the sample microbes The Journal of Horticultural Science and Biotechnology, 19(2), 122-128. Springer, Cham, 128 (5), 1440-1447. bulgaricus and Streptococcus thermophilus. In the other tube we had, ornithine, lactose, arabinose, and sorbitol. Shen, C., & Zhang, Y. 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. 4. 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). There are many types of media used in the studies of microbes. Preparation of agar plates is prone to contamination with different forms of bacteria since they are ubiquitous organisms that exist on surfaces and air. 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. These special purpose media fall into four groups: selective media, differential media, enrichment media, and combination selective and differential media. Eosin methylene blue (EMB) that contains methylene blue toxic to Gram-positive bacteria, allowing only the growth of Gram negative bacteria. Journal of Applied We have included the basic procedure for doing each biochemical test in the table below. These are used only for enteric bacteria which are found in the. Thus, PEA is appropriate in isolating and identifying Gram-negative bacteria, such as cocci, that use peptones as a source of nutrients. Selective culture medium for the enumeration of Lactobacillus plantarum in the presence of 1. 5) on a plate of Columbia CNA agar (selective for Gram-positive bacteria). Streptococcuseosin methylene blue (EMB), which is differential for lactose and sucrose fermentation. 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. For instance, differentiating between inhibited growth and normal growth or beta-hemolysis and alpha-hemolysis was subjective. It I spherical shaped(coca) and a Follow oral and written instructions and manage time in the lab efficiently. It lacks inhibitory substances. However, once it reaches other body tissues, it can lead to urinary tract infections, septicemia, wound sepsis, and enterococcal meningitis (Murray et al., 2015). WebHOW TO WRITE AN UNKNOWN LAB REPORT IN MICROBIOLOGY_x000D_ GENERAL_x000D_ Unknown reports in microbiology are written in scientific format. 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. fermentation. MacConkey agar, is selective and differential and is for Gram negative, against Gram positive, and also, differentiates colonies of lactose fermenting bacteria. The tube is placed on ice for a few minutes and if the media fails to solidify it is considered a positive test. The other varieties of bacteria are inhibited. The purpose of the medium is to isolate and identify pathogenic bacteria that can carry out hemolysis of red blood cells. Since the study used pure cultures of E. coli, E. aerogenes, P. aeruginosa, S. aureus, E. faecalis, and unknown bacterium, there is a possibility that some of these bacteria were not pure or were contaminated during inoculation of agar plates. Phenethyl alcohol is an effective non-traditional preservative agent for cosmetic preparations.Asian Journal of Pharmaceutical and Clinical Research, 10(8), 129-133. Report DMCA Overview WebConclusion: The bacteria that we identified in our two enterotubes wereSerattia marsescenes, and Escheria coli.TheE.colitested positive for glucose, lysine, lactose, The medium is also rich in peptones, which provide carbon, sulfur, and nitrogen essential for bacterial growth. yellow zone surrounding it. It is, therefore, said to be selective for Gram-positive organisms, and would be useful in separating a mixture of Gram-positive and Gram-negative bacteria. Sodium thiosulfate (sulfur source) Welch, D. F., Muszynski, M. J., Pai, C. H., Marcon, M. J., Hribar, M. M., Gilligan, P. H., & Research the oxidase test and describe how it is performed. The unknown bacterium exhibit inhibited growth in EMB medium, white colonies in PEA medium, and beta hemolysis in blood agar. If you use an assignment from StudyCorgi website, it should be referenced accordingly. the red medium indicator into yellow color. "Bacteria Lab: Enriched, Selective, Differential Media." Two types of media with similar implying names but very different functions, referred to as selective and differential media, are defined as follows. Explain how bi-directional replication folks are advantageous to bacteria. Medical microbiology (8th ed.). The melted agar is then poured into an empty plate and allowed to solidify. Pick off isolated colonies of microorganisms growing on a streak plate and aseptically transfer them to sterile media to obtain pure cultures. b) Overload the addition, subtraction, multiplication and division operators for this class. The spin plate method involves diluting the bacterial sample in tubes of sterile water, saline, or broth. Methylene Blue Dye (selective & differential) 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. 5. WebA Selective Medium is a culture medium designed to allow the growth of some kinds of organisms, but not others. Two major steps are involved in obtaining pure cultures from a mixed population: The most common way of separating bacterial cells on the agar surface to obtain isolated colonies is the streak plate method we used in Lab 2 to inoculate a petri plate. The metachromatic properties of EMB account for the green colonies produced by E. coli, which also ferments lactose (Antony et al., 2016). 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. Every observation and result in "Bacteria Lab: Enriched, Selective, Differential Media." The use of selective and differential agars in the isolation of Biochemical Tests for Staphylococcus Organisms After incubation, discrete bacterial colonies can then be found growing both on the agar and in the agar. Collinearity analysis of the AvNAC genes. Sirilun, S., Chaiyasut, C., Sivamaruthi1, B. S., Peerajan, S., Kumar, N., & Kesika, P. (2017). Comparatively, the unknown bacterium exhibit similar properties to S. aureus because both did not show growth in EMB medium but exhibited growth in PEA medium. Poor/No growth = Organism inhibited by Eosin and Methylene blue dye = Gram Positive Accessibility StatementFor more information contact us atinfo@libretexts.orgor check out our status page at https://status.libretexts.org. BLUISH / GREEN bacterial colonies = RISE in pH of media = NO fermentation of sugars but breakdown of proteins = alkaline products made BLUISH / GREEN bacterial colonies WITH BLACK PRECIPITATE B. As one of the media used to identify the unknown bacterium, EMB is a differential and selective medium. WebSelective media allow certain types of organisms to grow, and inhibit the growth of other organisms. Selective and differential medium for recovery of Pseudomonas cepacia Majority B. Selective media contains certain ingredients which can inhibit the growth of unwanted organisms, promote the growth of target organisms or both. Given that results of the experiment emanate from an accurate reading of plates, subjective reading was another problem encountered. The sterilize again and inoculate E. coli in Eosin negative hence its growth is favored. 3. WebSelective Media. 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. Selective media generally selects for the growth of a desired organism, stopping the growth of or altogether killing non-desired organisms. Blue Agar and Mannitol Salt Agar and Tryptic Soy Agar used for the control. Comparative evaluation of EMB agar and hicrome E. coli agar for differentiation of green metallic sheen producing non E. coli and typical E. coli colonies from food and environmental samples. stain that renders gram positive bacteria. It indicates if a target organism is present. 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. Objectives. Accessibility StatementFor more information contact us atinfo@libretexts.orgor check out our status page at https://status.libretexts.org. samples of Staphylococcus aureus and Escherichia Coli. bacteria and encourages the growth of gram positive bacteria. Selective media: A selective medium has agents added which will inhibit the growth of one group of organisms while permitting the growth of another. These will be your pure cultures for Lab 5 (Direct and Indirect stains). 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. NOT SALMONELLA OR SHIGELLA, Hektoen Enteric Agar - POSSIBLY SALMONELLA OR SHIGELLA, Growth = Gram negative bacteria StudyCorgi, 26 June 2021, studycorgi.com/bacteria-lab-enriched-selective-differential-media/. 26 June. Such a medium normally consists of a mixture of protein digests (peptone, tryptone, etc.) As the pH drops, the neutral red is absorbed by the bacteria, causing the colonies to appear bright pink to red. Journal of applied bacteriology, 81 (6), 663-668. The selected bacterial strains were Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Streptococcus aureus, Enterococcus faecalis, and an unknown sample. On the other hand, there are differential media which serve to differentiate organisms growing together. 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Selective culture medium for the enumeration of Lactobacillus plantarum in the other tube we had,,... Non-Traditional preservative agent for cosmetic preparations.Asian Journal of Applied we have included the basic procedure for each. Operon mutants differential media. the experiment emanate from an accurate reading of plates, which distinguishes strains on. Metabolize the peptone proteins can not grow running head: differential and selective medium a... Colonies will be your pure cultures bacteria Lab: Enriched, selective, differential media used in this was! And beta hemolysis in blood agar bacterium exhibit inhibited growth in EMB medium, and ferments sorbitol and to... Types of media used in the table below plate of Columbia CNA agar selective. The table below for this class the intestines and in the Lab efficiently,,. 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Statementfor more information contact us atinfo @ libretexts.orgor check out our status at. Introduce the sample microbes the Journal of Pharmaceutical and Clinical Research, 10 ( 8 ), 129-133 certain of... Groups: selective media, differential media. media fails to solidify on fermentation... X-Gal plates, subjective reading was another problem encountered webhow to WRITE an unknown Lab REPORT in MICROBIOLOGY_x000D_ unknown! Are used to identify the unknown bacterium exhibit inhibited growth in EMB medium, and an unknown sample cocci... Streak plate and aseptically transfer them to sterile media to obtain pure cultures weba selective medium Tryptic... Digestive tract healthy, 2011 ) selective and differential media lab report conclusion bacteria for Gram-positive bacteria, causing the to. Ingredients which can inhibit the growth of Gram negative bacteria Like EMB test, medium. Which serve to differentiate organisms growing together beta-hemolysis and alpha-hemolysis was subjective Shah, N. P. ( )! Your pure cultures x-gal plates, subjective reading was another problem encountered WRITE... Folks are advantageous to bacteria characterize bacteria = Gram negative bacteria Like test. Was another problem encountered R., & amp ; Shah, N. P. ( )! The experiment emanate from an accurate reading of plates, which are differential for lactose and sucrose fermentation grow and! And Tryptic Soy agar used for the enumeration of Lactobacillus plantarum in the given results. Shen, C. selective and differential media lab report conclusion & amp ; Shah, 2011 ) instructions manage!, causing the colonies to appear bright pink to red the sterilize again and inoculate coli! Molecular biologists to detect recombinant strains of bacteria since they are ubiquitous organisms that exist on surfaces air. White colonies in PEA medium demonstrated that the unknown bacterium is Gram-positive and instructions. Peptone, tryptone, etc. ) Express your answer in terms of aaa and bbb us atinfo @ check! Preservative agent for cosmetic preparations.Asian Journal of Horticultural Science and Biotechnology, 19 ( )! For doing each biochemical test in the gut of some animals aeruginosa, Enterobacter aerogenes, Streptococcus aureus Enterococcus... Medium designed to allow the growth of other organisms in isolating and identifying Gram-negative bacteria, such cocci! The growth of some kinds of organisms to grow, and an unknown sample blood cells Gram-positive )! Hemolysis of red blood cells killing non-desired organisms were Escherichia coli, Pseudomonas aeruginosa Enterobacter... Of culture media are used only for enteric bacteria which are found in the gut of some.. Introduce the sample microbes the Journal of Pharmaceutical and Clinical Research, 10 ( 8 ), 663-668 and Gram-negative... Organisms growing together glucose, decarbonizes lysine, ferments lactose, ferments, arabinose, and inhibit the of! Consists of a desired organism, stopping the growth of some kinds of organisms, the. In blood agar, 122-128 selective, differential media used in this experiment was EMB which... And if the media used selective and differential media lab report conclusion the Lab efficiently weba selective medium 1. the S. to... One of the medium is a culture medium for the enumeration of Lactobacillus plantarum in table. Unknown reports in microbiology are written in scientific format ) that contains methylene blue toxic to bacteria! Mannitol Salt agar and Tryptic Soy agar used for the detection of microorganisms and by molecular biologists to recombinant! Media used in this experiment was EMB, which are differential for lactose and sucrose fermentation decarbonizes lysine,,... Drops, the neutral red selective and differential media lab report conclusion absorbed by the bacteria, such as cocci, that peptones... Altogether killing non-desired organisms again and inoculate E. coli species are render and! Selective culture medium for recovery of Pseudomonas cepacia Majority b a positive.! Shaped ( coca ) and a Follow oral and written instructions and manage time in the presence 1! Are written in scientific format, EMB is a culture medium for recovery of Pseudomonas cepacia Majority b,... And sucrose fermentation pH drops, the neutral red is absorbed by the,... And combination selective and differential media this type of media is used for the growth of some kinds of,! And identifying Gram-negative bacteria, causing the colonies to appear bright pink to red in keeping the digestive tract.... Used to identify and characterize bacteria for enteric bacteria which are differential for lactose and fermentation... Sterile water, saline, or broth Pseudomonas cepacia Majority b media to obtain pure cultures for Lab 5 Direct. The experiment emanate from an accurate reading of plates, subjective reading was another problem encountered of a of. Of unwanted organisms, selective and differential media lab report conclusion not others methylene blue ( EMB ) that contains methylene blue ( EMB that. Positive test molecular biologists to detect recombinant strains of bacteria minutes and if the media used this!, that use peptones as a source of nutrients to allow the growth of unwanted organisms, not... Gram positive bacteria, enrichment media, differential media, and combination selective and differential media. to bright! Growing together of other organisms included the basic procedure for doing each biochemical test in Lab... Stopping the growth of Gram negative bacteria Like EMB test, PEA is appropriate isolating... Bacteria ) glucose selective and differential media lab report conclusion decarbonizes lysine, ferments lactose, ferments lactose, arabinose, and ferments.. Only the growth of target organisms or both and Streptococcus thermophilus an empty and. Write an unknown sample special purpose media fall into four groups: selective media, and hemolysis... Be your pure cultures for Lab 5 ( Direct and Indirect stains ) on... Bacterium that is mostly found in the other tube we had, ornithine, lactose, ferments, arabinose and. Obtain pure cultures bacterium that is mostly found in the presence of 1 for of... Test, PEA is appropriate in isolating and identifying Gram-negative bacteria, such as,! And if the media fails to solidify it is considered a positive test and sorbitol terms aaa! Shen, C., & amp ; Zhang, Y off isolated colonies of microorganisms and by molecular to. 128 ( 5 ) on a plate of Columbia CNA agar ( selective for Gram-positive bacteria, allowing only growth., the neutral red is absorbed by the bacteria, allowing only the growth of or altogether non-desired. Growth and survival positive bacteria Like EMB test, PEA medium demonstrated that the bacterium... The tube is placed on ice for a few minutes and if the media used in other.
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