2002). Only those with compromised immune systems are thought to be susceptible to an infection.DisinfectionM. The genome of the Micrococcus is rich in guanine and cytosine, they typically exhibit about 65 to 75% GC-content. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. (2) Micrococcus spp. Its genome has a high G + C content. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. Growth can produce colonies of a certain size and consistency, as well as pigmentation. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . The bacterium also colonizes the mouth, mucosae, oropharynx, and upper respiratory tract. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. M. luteus tests positive for catalase, oxidase, utilization of D-glucose, sucrose and D-mannose, and has A2 type peptidoglycan that contains L-lysine as the diagnostic amino acid, MK-8 and MK-8 (H 2) are the major menaquinones (Stackebrandt et al. The configurations are the result of the plane of cellular division exhibited by the organism. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). Streptococcus spp. 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. 2. They are found in many other places in the environment, including dust, water, and soil. 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. They are seldom motile and are non-sporing. The Micrococcus genus is known to be found on dust particles, in water, on skin and skin glands in vertebrates, and some species can be found in milk. Micrococcus luteus was found gram-positive cocci, non-spore forming, non-haemolytic, non-motile aerobic or facultative anaerobes, arranged in pairs, irregular clusters and tetrads. Exposure to these wavelengths of ultraviolet light has been correlated with an increased incidence of skin cancer, and scientists believe this pigment can be used to make a sunscreen that can protect against ultraviolet light.[9]. I then repeated this three more times to further purify the isolate. This microbe forms large, round colonies. Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. For example, M. luteus and M. lylae are 40-50% genetically different. Micrococcus luteus is a coccus shaped gram-positive bacteria. M. luteus oxidize carbohydrates to form carbon dioxide and water, and it does not produce acid from the glucose as well as it does not make arginine dihydrolase or b-galactosidase. I then streaked the exposed swab onto a petri dish to allow the organisms to grow, incubating them at 25 degrees Celsius in a dark cabinet. . Recently, as an opportunistic pathogen, this organism was recognized and it has been implicated in recurrent bacteremia, septic arthritis, septic shock, endocarditis, intracranial suppuration, meningitis, and cavitating pneumonia in immunosuppressed patients. [3] Its genome was sequenced in 2010 and is one of the smallest genomes of free-living Actinomycetota sequenced to date, comprising a single circular chromosome of 2,501,097 bp.[4]. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. To sample, I used sterile cotton swabs and streaked them on TSA plates. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. It grew in circular, entire, convex and creamy yellow pigmented colonies heaving 0.5-2.5 diameter. Some of the Micrococcus are pigmented bacteria, for example, M. roseus produces reddish colonies and M. luteus produces yellow colonies. 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. Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. It is Gram positive, Coagulase positive, catalase and oxidase positive non-motile bacteria. If the agar plate is held up to a light source, you can sometimes see through the agar, as is pictured on the left. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. Pearls of Laboratory Medicine 1995; Wieser et al. Micrococci can often carry plasmids that provide the organism with useful traits. 2019). All pathogenic strains of S. aureus are coagulase positive whereas the nonpathogenic species (S. epidermidis for example) are coagulase negative. The catalase test tests to see if the microbe contains catalase. The colony forms as a yellow, shiny round blob. I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. [7], In 2003, it was proposed that one strain of Micrococcus luteus, ATCC 9341, be reclassified as Kocuria rhizophila. It is a gram positive, coccus shaped microbe, and contains catalase. U.S. 2022 American Association for Clinical Chemistry. Enter the email address you signed up with and we'll email you a reset link. It is reported here that gliotoxin selectively spares a unique class of haemopoietic stem cell that forms large (HPP) colonies in the presence of mixtures of MCSF and IL3. As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. Though today the immuno-compromised patients take the risk of the infection that has grown. // Source publication +9 Isolation and Identification of Potential Pathogenic Bacteria in Living Carp (Cyprinus carpio Linnaeus, 1758) Sold in. This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-). M. luteus has one of the smallest genomes of actinobacteria with a circular chromosome of 2,501,097 base pairs and a GC content of 73%. They are considered as normal comensal of human skin and upper respiratory tract. 2014). Continuing without changing cookie settings assumes you consent to our use of cookies on this device. Although many of the members of these families have low pathogenicity and are classified as causes of opportunistic infections, those organisms most often cited as pathogens are listed on this slide. This microbe is coccus shaped and forms in tetrads. The streptococci and enterococci are categorized by expression of either beta, alpha, or gamma hemolysis on sBAP, depending on the genus/species. Reduction of O2 produces superoxide radicals and hydrogen peroxide, or the hydroxyl radical. 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. Bacillus megaterium fermented sugars but didn't grow in the anaerobic area of the butt. In the case of a negative test, no colour change is observed. Colonies typically have a lemon-yellow pigment. I also did a fluid thioglycolate test to determine the bacterias oxygen class. Once the culture was deemed pure enough, I inoculated a slant tube. "EPR and ENDOR detection of compound I from Micrococcus lysodeikticus catalase". I suspect that the API 20E test strip results were negated by having used an inactive colony, as the fluid thyoglycate test failed as well. Micrococcus luteus can synthesize the iso-branched and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters. 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. Micrococcus luteus (left) Alcaligenes faecalis (middle) Reduction of nitrate to nitrite to be used as a final electron acceptor/Nitrate reductase. Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. Enterococcus faecalis. Intermediates or end products of these varied metabolic activities can be detected by performing biochemical assays on a bacterial culture. Simple biochemical tests like the one above have always been an important aid to identification of bacteria, because the different bacterial groups and species have characteristic metabolic activities. SUMMARY: 1250 Gram-positive and catalase-positive cocci were isolated from bacon, pig and human skin and dust, and their morphology, physiology and biochemical characters examined. From four species that have been identify, only Bacillus subtilis and Pseudomonas diminuta that show ability to block pathogen bacteria Edwardsiella tarda. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. Micrococcus Catalase Test: The catalase test facilitates the detection of the enzyme catalase in bacteria. Genus: Micrococcus. Micrococcus luteus was formerly known as Micrococcus lysodeikticus. For the micrococci, susceptibility to the antimicrobial agents, bacitracin and furazolidone, as well as the modified oxidase test can be performed to distinguish this group from the staphylococci. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. The species Micrococcus luteus (Schroeter 1872) Cohn 1872 (Approved Lists 1980) emend. Bharath. Biochemical Test and Identification of Staphylococcus aureus. The identification of three different biovars within the species M. luteus has the advantage that the three groups can be differentiated without nomenclatural changes having to be introduced. On this Wikipedia the language links are at the top of the page across from the article title. Staphylococcus spp. When looking back at all the data, it is fairly likely that the isolate is Micrococcus luteus, especially when looking at the fluid thyoglycate test, the colony morphology, and the antibiotic susceptibility. They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. 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. Some streptococci also possess unique cell wall carbohydrate antigens that can be identified by reactivity with specific indicator antibodies in an agglutination assay (Lancefield typing). (2019, March 14). "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. A summary table is included which lists the results of the identification criteria discussed in the tutorial. They are positive for catalase and oxidase ( modified). Your email address will not be published. 1 Nevertheless they have been documented to be causative organisms in cases of bacteremia, endocarditis, ventriculitis, peritonitis, pneumonia, endophthalmitis, keratolysis and septic arthritis. 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). Another set of tests that I performed were physiological tests, including testing for oxygen class with fluid thyoglycate, checking for catalase using hydrogen peroxide, checking for cytochrome c oxidase using oxidase test strips, and using an API 20 E test strip to test for various different physiological traits such as Sulphur reduction, which had 20 different tests. 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. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. M. luteus can also cause septic shocks in immuno-compromised people. . At U.S. Micro Solutions, our microbiology laboratory is a licensed, accredited source for thorough and accurate environmental testing . Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar. These residential skin organisms are also fairly ubiquitous in the environment. 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). There have been several deaths in immuno-compromised children that are caused by. Hybridization studies indicate that species within the genus Micrococcus are not closely related, showing as little as 50% sequence similarity. Thank you for joining me, Karen Krisher, on this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. M. luteus present on the human skin can transform compounds present in sweat into compounds with an unpleasant odour. Micrococcus A study with 188 micrococci, identified only to the genus level, demonstrated MICs at achievable concentrations for most -lactams, aminoglycosides, glycopeptides, clindamycin and the most active drug in vitro, rifampin. As mentioned, the type of hemolytic reaction produced on sBAP is a major clue toward the identification of the genus. This presentation will focus on the laboratory tests useful for the differentiation among the families as opposed to the more complicated differentiation and identification of the organisms within the different genera. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. Micrococcus luteus is another gram-positive coccus that also is catalase positive. The pathogen, Staphylococcus aureus, is notably coagulase-positive while most other members of the family are coagulase-negative. Enterococcus spp. I grew my isolated in Tryptic Soy Broth (TSB) for a week to prepare for DNA extraction. In Wikipedia, The Free Encyclopedia. M. luteus is the majority of reads on the species level. Catalase is an enzyme produced by the organism to neutralize the toxic effects of hydrogen peroxide in the environment of the cell. Microbiome 2(27). Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. We were aiming to isolate a bacterium from this initial environmental sample by using repeated quadrant streaks to isolate single species colonies, therefore producing a pure culture that we could do a multitude of tests on which include Gram staining, genetic analysis and antibiotic testing along with many other tests. As its name suggests, mannitol salt agar (MSA) contains 1% mannitol (sugar), 7.5% salt, and agar as a solidifying agent. M. lylae can be distinguished from the closely related species Micrococcus luteus by lysozyme susceptibility, genetic composition, and the type of cell-wall peptidoglycan. You can change these settings at any time, but that may impair functionality on our websites. They contain cytochromes and are resistant to lysostaphin. The catalase test also indicated that the microbe does not have catalase, despite the metagenomic binning test suggesting it. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). Obtain a glucose fermentation tube. 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. This can occur due to the presence of a reduced number of, Most of the Micrococcus infections are discovered through the process of elimination along with the presence of the abundant Micrococcus tetrads in the lesions or cysts. The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract. In the last control, agar-agar was inoculated with 1 mL of M. luteus washed live cells and incubated for 24 h at 37 C. M. luteus has one of the smallest genomes of all the bacteria. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. They occur in pairs, tetrads or clusters but not in chains. The optimum growth temperature is 25-37C. They are normal inhabitants of the human body and may even be essential in keeping the balance among the various . Date: FEB.11.2016 These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium, M. luteus played an important part in Flemings discovery of, 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. When using a fluid thyoglycollate test it resulted in the isolate being a strict aerobe, with all of the bacterium being at the top of the medium where it is oxygenic. Micrococcus spp. Like MSA, this medium also contains the pH indicator, phenol red. The oxidase test results suggest that the microbe does not contain oxidase, despite what the metagenome binning test showed. Recent reports, however, confirm that micrococci may be associated with human infections, particularly in immunosuppressed patients. When viewing the organism microscopically, the staining characteristics of the organism provide important information for identification. The colony morphology of being yellow, shiny and smooth line up perfectly with M. luteus (Public Health England). Micrococcus are aerobic, Gram-positive cocci ranging in size from 0.5 to 2.0 m in diameter. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. 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. If acid is produced, the pH indicator will turn yellow. 2-9 In these reports micrococci were identified on the basis of their . My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. The MacConkey agar is selective for gram-negative which is why my microbe didnt show much growth on it, and because it didnt change colors it means it didnt ferment the lactose. Micrococcus luteus was one of the early examples of novel codon usage,[5][6] which led to the conclusion that the genetic code is not static, but evolves. Sarah D. Perkins., Government of Canada, Public Health Agency of Canada. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . They can even survive in conditions that have very little of even no oxygen, using glycolysis and alternative electron acceptors to create ATP. This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. 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. The colonies are a pale, translucent yellow, and are shiny when looked at in the light. The previous classification of these organisms was much simpler; however, with the use of more advanced genetic sequencing methods, these gram-positive cocci have expanded into 4 separate families and their associated genera. Where the M. luteus can cause severe skin infections and is sometimes clinically mistaken for Staphylococcus aureus. These can indicate that the genus of the Micrococcus is more closely related to the genus Arthrobacter than it is to other coccoid genera such as Staphylococcus and Planococcus. Wikipedia contributors. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. They grow in circular, entire, convex, and usually non-pigmented or cream white colonies with diameters of approximately 4 mm after 2-3 days on the plate at 37C. The nasal cavity microbiome primarily consists of the phyla Actinobacteria, Firmicutes and Proteobacteria (Bassis et al. It had 27,372 contigs in assembly. Environmental Testing. 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. 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). 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. In conclusion, some of my results were inconclusive and conflicting. This is in agreement with a study by Mohana et al., (2013) who reported that the carotenoid pigments had antioxidant properties of Micrococcus luteus was reported at IC50 of 4.5mg/ml, also the .