Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. Micrococcus Luteus Oxidase Test: The microdase test, also known as modified oxidase test is a rapid test to differentiate Staphylococcus from Micrococcus which are Gram-positive cocci possessing catalase enzymes. We found this to be true because the filter paper turned blue, which showed that the species has the cytochrome c oxidase enzyme. The organism itself is susceptible to almost all drugs, with a few strains being resistant to nitrofurantoin, macrolides and lincomycin (Public Health Canada). Although the Micrococcus rarely causes infections or problems in the body, with the compromised immune systems, it has been known to get certain skin infections that are caused by Micrococcus luteus. They can be found in boiling hot springs; they can be found in frigid waters in the artic. Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher On this Wikipedia the language links are at the top of the page across from the article title. Intermediates or end products of these varied metabolic activities can be detected by performing biochemical assays on a bacterial culture. All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. Enterococcus spp. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. Colony morphology, the elucidation of the characteristics of the growth of the bacterium on agar medium, in most cases trypticase soy agar containing 5% sheep blood (sBAP). While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. The M. luteus genome encodes about four sigma factors and fourteen response regulators, a finding indicative of the adaptation to a rather strict ecological niche. Continuing without changing cookie settings assumes you consent to our use of cookies on this device. Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. Micrococcus luteus biochemical test result. All three types of hemolytic reactions are represented on this slide. Micrococcus luteus is another gram-positive coccus that also is catalase positive. Though today the immuno-compromised patients take the risk of the infection that has grown. Micrococcus species are oxidase-positive, where these can be used to distinguish them from some other bacteria like most of the Staphylococcus species, which are generally oxidase-negative. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. In contrast to staphylococci, usually, penicillin is sensitive. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). Three bacteria, E.coli, Staphylococcus epidermis, and Micrococcus luteus, have been grown on TSI or Triple sugar iron agar medium, which is a . Other articles where Micrococcus luteus is discussed: Micrococcus: found in milk, such as M. luteus, M. varians, and M. freudenreichii, are sometimes referred to as milk micrococci and can result in spoilage of milk products. November 27, 2021 at 9:14 AM Hi sir, why Macrococcus and Micrococcus will not ferment . [7], In 2003, it was proposed that one strain of Micrococcus luteus, ATCC 9341, be reclassified as Kocuria rhizophila. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. Micrococci are microbiologically characterized as gram-positive cocci, 0.5 to 3.5 m (micrometres; 1 m = 10-6 metre) in diameter. The conflicting results of the metagenome binning and the catalase test influenced this mistake. The differentiation is based on the detection of oxidase enzyme. They contain cytochromes and are resistant to lysostaphin. The M. luteus genome encodes about four sigma factors and fourteen response regulators, a finding indicative of the adaptation to a rather strict ecological niche. The oxygen class of the microbe, obligate aerobe, matches up with the predictions I had made about it because the bacteria was originally sourced in a nostril. 2- M. luteus 3- M.roseus Culture: - Strictly aerobic at 37C incubation (24 hr) - Grow on ordinary media Nutrient agar - Blood agar and on the Blood agar M. roseus (pink) M. luteus (white) M.varians (yellow) - On m annitol salt agar grow given r ise to rose or pink colonies except M. roseus. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. The confidence on that reading is decently confident, evidenced by the blue coloring. To sample, I used sterile cotton swabs and streaked them on TSA plates. The categorization of bacteria in the clinical laboratory is based on the reactivity of the organism when stained by the Grams staining reagents. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. M. luteus oxidizes carbohydrates to CO2 and water, and it does not produce acid from glucose as well as it does not make arginine dihydrolase or b-galactosidase. Wieser et al. Required fields are marked *. 1995; Wieser et al. Coagulase production is used as a virulence factor by the organism. The negative oxidase result conflicts with M. luteus (Public Health England). CCTC (English) In the case of a negative test, no colour change is observed. Many of the tests did line up with M. luteus though, such as the fluid thyoglycate test, which showed that it was an obligate aerobe. The differentiation is based on the detection of the oxidase enzymes. Filter paper disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride (oxidase reagent) in dimethyl sulfoxide (DMSO) are used. Micrococcus Public Health Agency of Canada., Public Health England. I used an oxidase test strip and water to determine if cytochrome C oxidase was present, and performed a catalase test to determine if catalase was present. Pearls of Laboratory Medicine The differentiation is based on the detection of the oxidase enzymes. Of those only Vancomycin was closer than 50 millimeters, being 38 millimeters. Thus, when dealing with Micrococcus infections, it usually takes several cultures that are being grown and examined before one realizes that Micrococcus is indeed the culprit. Lastly, do not underestimate the variety of different genera in the four families of the Gram- positive cocci that can, under the right circumstances, move from relatively harmless saprophytes to disease-producing opportunists. Micrococcus is a spherical bacterium found on dead or decaying organic matter while Staphylococcus is a gram-positive bacterial genus that produces a bunch of grape-like bacterial clusters. The laboratory assay is performed by mixing a sample of the bacterial colony with a drop(s) of hydrogen peroxide placed on a microscope slide. Micrococcus luteus biochemical test result. Production of bubbles indicates a positive reaction. Catalase, if present, will break down the hydrogen peroxide to water and oxygen. Micrococcus are aerobic, Gram-positive cocci ranging in size from 0.5 to 2.0 m in diameter. Microbiome 2(27). Micrococcus luteus (left) Alcaligenes faecalis (middle) Reduction of nitrate to nitrite to be used as a final electron acceptor/Nitrate reductase. It is an aerobic exopolysaccharide producing bacteria that may be found on the skin, mouth and sometimes in the throat of mammals 13,14. Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. They occur in pairs, tetrads or clusters but not in chains. Bharath. Biochemical Test and Identification of Staphylococcus aureus. They usually can occur in the irregular clusters, tetrads, and in pairs, where the individual cells are about one to 1.8 mm in diameter and these are usually non-motile and non-spore-forming in nature. The EMB plate is also selective for gram-negative bacteria which is probably why the bacteria didnt grow on it. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . A positive test leads to the development of blue or purple-blue on the disc within two minutes. The genome of the Micrococcus is rich in guanine and cytosine, they typically exhibit about 65 to 75% GC-content. Note the bright yellow, non-diffusable colony pigment which is a defining characteristic of M. luteus. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. Defining characteristics of Micrococcus are the ability to aerobically produce acid from glucose glycerol, aesculin hydrolysis, arginine dihydrolase, major pigment production, motility, and conversion of nitrate to nitrite. They are found in many other places in the environment, including dust, water, and soil. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. A rare case of prosthetic valve endocarditis is caused by Micrococcus luteus, it is described and compared with the few cases that are reported in the literature, as well as the clinical features, microbiological profile, therapy, and prognosis of common prosthetic valve endocarditis. Its genome has a high G + C content. (2019, April 4). There are around nine species that are recognized in the genus. 2023 Universe84a.com | All Rights Reserved, Blog: Microbiology and infectious disease, Anti-Mullerian Hormone (AMH) Test: Introduction, Result, Unit, Normal Range, Test Method, Clinical Significance, and Keynotes, Anti -TPO Antibody: Introduction, Test Result, Unit, Normal Range, Assaying Method, and Keynotes, HPV Genes detection using Real-Time PCR: Introduction, Principle, Test Requirements, Procedure, Result Interpretation and Keynotes, Microbiology Reporting Techniques: Introduction, List of Templates, and Keynotes, Acetamide Utilization Test: Introduction, Composition, Principle, Test Requirements, Procedure, Result-Interpretation, Limitations, and Keynotes, https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/887570/UK_SMI_ID_07i4.pdf, https://en.wikipedia.org/wiki/Micrococcus_luteus, https://europepmc.org/article/med/14576986, https://www.ajicjournal.org/article/S0196-6553(13)01146-2/fulltext. Alcaligenes faecalis Bacillus megaterium . They are generally strict aerobes and these can generally reduce nitrate concentration. There are 3 biovars of this organismand they possess quite diverse chemotaxonomic features with respect to their menaquinone systems, cell-wall compositions, and Fourier transform-infrared (FT-IR) spectroscopy patterns, as well as biochemical properties. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. The Bacterial Genomic DNA Isolation Kit provides convenient and simple step-by-step method for isolating quality genomic DNA from gram-negative and gram-positive bacterial species. Rawis M, and AK Ellis (2019). Staphylococcus aureus. Micrococcus has a substantial cell wall in which it may comprise as much as 50% of the cell mass. The fluid thioglycallate test showed that the bacteria was an obligate aerobe because the growth was concentrated at the top of the tube in the pink region. The Gram stain, while it was gram variable, does not ideally match with the genetic test that resulted in Micrococcus luteus, which can be gram variable but is usually gram positive (Bonjar). I isolated the DNA by lysing the cell and using the Power Sol DNA Isolation Kit to isolate the DNA from the other parts of the cell. While defining the characteristics of the Micrococcus are the ability to produce acid aerobically from the aesculin hydrolysis, glucose glycerol, arginine dihydrolase, motility, major pigment production, and conversion of nitrate to nitrite. . It did not reduce sulphur, digest gelatin, or reduce nitrate, among some of the more interesting tests, which all disagree with M. luteus. This would be read A/NC. 900 Seventh Street, NW Suite 400 The taxonomic assignment of this microbe was micrococcus luteus because it was the only bin that PATRIC gave. Organisms commonly found on the body include Micrococcus, Staphylococcus, and Corynebacterium species (Dermnet New Zeland). luteus is capable of surviving in environments "Pathogen Safety Data Sheets: Infectious Substances Micrococcus spp", "Genome sequence of the Fleming strain of Micrococcus luteus, a simple free-living actinobacterium", "Role of GC-biased mutation pressure on synonymous codon choice in Micrococcus luteus, a bacterium with a high genomic GC-content", "Reclassification of ATCC 9341 from Microccus luteus to Kocuria rhizophila", https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=1132534318, Articles with unsourced statements from January 2023, Creative Commons Attribution-ShareAlike License 3.0, This page was last edited on 9 January 2023, at 09:24. In the case of a negative test, no colour change is observed. Methods: To start isolating the bacterium, I used a sterile swab wet with deionized water provided by the lab to pick up bacteria from the shower drain. M. luteus has one of the smallest genomes of all the bacteria. These residential skin organisms are also fairly ubiquitous in the environment. // Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. I hypothesized that it would be a bacteria commonly found in the nasal cavities and likely from the aforementioned phyla, so it would likely do best living in an aerobic, humid, and warm environment. Streptococcus spp. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). nishinomiyaensis and Micrococcus luteus. Similar to the situation in the prosthetic valve endocarditis that is caused by more aggressive and highly resistant bacteria. This is a test commonly used when trying to identify Gram-negative enteric bacteria, all of which are glucose fermenters but only some of which produce gas. In order to determine physiological characteristics of the culture such as cell shape, arrangement, and whether it was gram positive or negative, which helps determine the cell wall type of the microbe, I performed a gram stain. View Online Copy of Unknown ID Chart.xlsx from BIOL 250L at University of South Carolina. Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. Enterobacter aerogenes fermented the sugars but turned to the amino acids. Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. It is a gram positive, coccus shaped microbe, and contains catalase. Environmental Testing. Coagulase Results: The microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. The staphylococci and micrococci are catalase producers while the streptococci, enterococci, and a variety of other Gram-positive cocci are catalase-negative. I grew my isolated in Tryptic Soy Broth (TSB) for a week to prepare for DNA extraction. AACC uses cookies to ensure the best website experience. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. The catalase test also indicated that the microbe does not have catalase, despite the metagenomic binning test suggesting it. The gram stain of this microbe showed that it is gram positive because it stained purple. However, the most promising antibiotic regimen that is proposed for the treatment of the Micrococcus luteus seems to be a combination of amikacin, vancomycin, and rifampicin. View Biochemical Tests Lab Report.docx from BIOL 240 at University of Alaska, Anchorage. Genus: Micrococcus. Micrococci can often carry plasmids that provide the organism with useful traits. 91 988-660-2456 (Mon-Sun: 9am - 11pm IST), Want to read offline? Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) This lines up with M. luteus resistances from the tests. Gram-positive organisms appear purple under the microscope as opposed to the other category of bacterial cell with its own unique cell wall, the Gram-negative rod or coccus, which stains red. Biochemical Pr Organism Enterococcus faecalis Lactococcus lactis Micrococcus luteus Staphylococcus I would hypothesise that because we clean our shower regularly, that most of the bacterium un the shower drain would come from people in my family taking a shower, and mostly be from the skin. The MacConkey agar showed very little growth, and did not have a change in color, indicating that the microbe was gram positive and not a fermenter. 2014). Colonies typically have a lemon-yellow pigment. The API test strips lack of results suggests that the I used the wrong test strip, I probably needed to use the Staph test instead of the Strep test, because the Strep test is for when Catalase is absent, but there could have been catalase present. The oxidase test results suggest that the microbe does not contain oxidase, despite what the metagenome binning test showed. They are fairly small as well, usually about a millimeter in diameter and of a normal height. Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. . This technique is centered on the performance of the classic Grams stain. You can change these settings at any time, but that may impair functionality on our websites. Principle of Microdase (Modified Oxidase) Test The microdase test, also known as modified oxidase test is a rapid test to differentiate Staphylococcus from Micrococcus which are Gram positive cocci possessing catalase enzyme. In the presence of atmospheric oxygen, the oxidase enzyme reacts with the oxidase reagent and cytochrome C to form the coloured compound, indophenol indicated as blue or purplish-blue colouration on the disc after the introduction of the bacterial colony on the disc. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. Micrococcus luteus is found in lots of places including skin, soil, dust, water, air, mouth, mucosae, oropharynx, and upper respiratory tract of humans (Wikipedia, Micrococcus luteus, 2019). This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-). In the 1920s, Alexander Fleming described lysozyme as a bactericidal factor of human and other animal tissues and secretions.1 He also discovered and named a yellow bacterium, Micrococcus lysodeikticus (now M luteus), that was highly susceptible to lysozyme-mediated killing.After exploring the susceptibility of other bacteria to lysozyme, Fleming proposed that M luteus and certain other . Further tests that I would do would be testing how much heat resistance it has, the density of a broth suspended sample using a dilution series, test for more antibiotic resistances, and how well it can grow in antibacterial mediums and mediums of different pH levels. U.S. 2022 American Association for Clinical Chemistry. Date: FEB.11.2016 It is often larger thanStaphylococcus. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. Coagulase, which is an enzyme involved in the conversion of serum fibrinogen to fibrin, causes the clotting of plasma. This is termed alpha () hemolysis and results in a greenish color surrounding the colony. The pathogen, Staphylococcus aureus, is notably coagulase-positive while most other members of the family are coagulase-negative. Thank you for joining me, Karen Krisher, on this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. Hemolysis is the lysis of the sheep erythrocytes within the agar by bacterial toxins (hemolysins) that are produced by the different genera of Gram-positive cocci. Micrococcus luteus pigment (Yellow) GLUCOSE FERMENTATION TEST This test detects the ability of the bacterium to break down glucose to pyruvic acid. [3] The colony morphology of being yellow, shiny and smooth line up perfectly with M. luteus (Public Health England).