micrococcus luteus biochemical tests

Optimum growth temperature is 25-37 C. In Wikipedia, The Free Encyclopedia. AACC uses cookies to ensure the best website experience. Coagulase production is used as a virulence factor by the organism. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. Each genus lends itself to a separate tutorial. If I were to continue researching this isolate, I would redo the API 20 E test strip with a fresh, active culture to ensure that it can reduce nitrate, and also the oxidase test to ensure that it does have cytochrome c oxidase present, which it should according to Public Health England. Luteus is an obligate aerobe (medical. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. Then to actually isolate a bacterium, I chose a colony from the initial plate and did a quadrant streak of it to further isolate the bacterium, and then incubated it at 38 degrees Celsius for a week. 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. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. There have been several deaths in immuno-compromised children that are caused by. Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. I used Prokka Genome Annotation to identify which genes are present in the bacterium, Kraken Metagenomics to identify what the DNA reads correlated to on different taxonomic levels, and SPAdes Genome Assembler to determine how many contigs were produced from the reads that were produced. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. Environmental Testing. They are likely involved in the biodegradation of many other environmental pollutants or detoxification. Micrococcus luteus are Gram-positive, to Gram-variable, motile -non motile, that are 0.5 to 3.5 micrometers in diameter and usually arranged in tetrads or irregular clusters. The catalase test tests to see if the microbe contains catalase. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. They are catalase-positive and usually aerobic with strictly respiratory metabolism. They have a respiratory metabolism, often producing little or no acid from carbohydrates, and are usually halotolerant, growing in 5% NaCl. 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. Only the Micrococcus luteus species contained cytochrome c oxidase. After it was sequenced using an Illumina MiSeq at the UAF Core lab, I used the online program Base Space to analyze the reads that I got from sequencing. Date: FEB.11.2016 Pearls of Laboratory Medicine Micrococcus luteus, a commonly found member of the Micrococcaceae, produces round colonies which are convex (curving outward) with smooth, entire margins. Motility (Motile / Non-Motile) Non-Motile. The positive catalase result lines up with M. luteus (Public Health England). For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. Finally, when looking at the API 20E strip none of the results returned positive despite the culture being active and fresh from being streaked recently. This would be read as K/A. 1995; Wieser et al. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. When performing tests for physiological traits, the results were limited. Kaiju webserver metagenome binning analysis chart. Download Scientific Diagram from www.researchgate.net. . Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher With the advent of newer molecular techniques, genus and species confirmation is even faster and easier. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). 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 . 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). A number of sophisticated tools are now available for clinical diagnosis. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). For example, Micrococcus varians is now known as Kocuria varians. Micrococcus luteus ( M. luteus ), is a Gram-positive bacteria, 0.05 to 3.5 microns in diameter, that is most commonly found in mucous membranes such as the nasal cavities, the upper respiratory tract, and the lining of the mouth. It shows that it is mostly Actinobacteria, with some firmicules ,and proteobacteria mixed in (Figure 2). Figure 2. U.S. 2022 American Association for Clinical Chemistry. 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. They are indole negative, oxidase-positive, catalase-positive, and citrate negative. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. Micrococcus luteus | Taxonomy - PubChem Apologies, we are having some trouble retrieving data from our servers. The EMB plate is also selective for gram-negative bacteria which is probably why the bacteria didnt grow on it. The streptococci and enterococci are categorized by expression of either beta, alpha, or gamma hemolysis on sBAP, depending on the genus/species. They are generally strict aerobes and can generally reduce nitrate. This technique is centered on the performance of the classic Grams stain. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . I also hypothesize that it will be an aerobic organism, given that I found it in a well aerated environment and it has survived until I cultured it. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. 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. For the second control, 100 L of water sample dilutions were plated on agar-agar. Alpha-hemolytic colonies often assume a greenish hue. 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. 2. It shows that the sample contains bacteria from the Terrabacteria group. The optimum growth temperature is 25-37C. 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. They are generally strict aerobes and these can generally reduce nitrate concentration. It may appear in irregular clusters, groups of four or eight. 570 . You can change these settings at any time, but that may impair functionality on our websites. This lines up with M. luteus resistances from the tests. View Biochemical Tests Lab Report.docx from BIOL 240 at University of Alaska, Anchorage. Micrococcus Catalase Test: The catalase test facilitates the detection of the enzyme catalase in bacteria. 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. Micrococcus species occur in a wide range of environments, including dust, water, and soil. Organisms commonly found on the body include Micrococcus, Staphylococcus, and Corynebacterium species (Dermnet New Zeland). The categorization of the Gram-positive cocci based on their possession of the enzyme catalase, a quality which can be easily assayed in the lab. If acid is produced, the pH indicator will turn yellow. appear as agents of infection causing endocarditis. M. luteus has a G-C content of about 65-75 mol%. Depend on Gram stain Test and Biochemical Test find four species of cellulolytic bacteria such as Bacillus subtilis, Pseudomonas diminuta, Micrococcus luteus and Plesiomonas shigelloides. 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. This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. M. luteus can also cause septic shocks in immuno-compromised people. Rawis M, and AK Ellis (2019). Structure and PhysiologyThis bacteria is Gram-positive, spherical, and an obligate aerobe. They are positive for catalase and oxidase ( modified). They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. When looking at the antibiotic test results, the isolate is resistant to none of the applied antibiotics, and is only lightly to intermediately resistant to oxacillin. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. The catalase and the oxidase tests came up negative, because the catalase test did not form bubbles, and the oxidase test did not see a color change. These organisms range in virulence potential from commensal organisms found on the skin and mucous membranes to pathogens associated with serious infections. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. Its genome has a high G + C content. // Micrococcus luteus is another gram-positive coccus that also is catalase positive. Micrococcus luteus in tetrads arrangement. These residential skin organisms are also fairly ubiquitous in the environment. Staphylococcus spp. 2. It is an opportunistic pathogen, only pathogenic enough to cause disease in weakened immune systems (Medical Laboratories). This is termed alpha () hemolysis and results in a greenish color surrounding the colony. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar. Coagulase Results: Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. In future works with this microbe, I probably would want to purify the culture more and redo the tests. Micrococcus are aerobic, Gram-positive cocci ranging in size from 0.5 to 2.0 m in diameter. 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. They are seldom motile and are non-sporing. Although the initial placement of the Gram- positive cocci into broad categories is relatively easy, further classification of some isolates can be challenging and requires a battery of tests to arrive at a definitive identification. This microbe forms large, round colonies. I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. Staphylococcus aureus. Microbiome 2(27). Typical colonies are often described as gray to white, moist or glistening. For example, M. luteus and M. lylae are 40-50% genetically different. Micrococcus luteus Grown on BrainHeart Infusion Agar, Klebsiella characteristics on MacConkey Agar, Clinical Case Leukocyte Vacoulation Bacterial Infection, Segmented neutrophilic granulocyte during degradation, DIC (Disseminated intravascular coagulation), Creatinine Phosphate Kinase (CPK) and CK-MB Overview. Micrococci are usually not pathogenic. This microbe forms large, round colonies. The nasal cavity microbiome primarily consists of the phyla Actinobacteria, Firmicutes and Proteobacteria (Bassis et al. Micrococcaceae. Reduction of O2 produces superoxide radicals and hydrogen peroxide, or the hydroxyl radical. Perkins1, Sarah D., Jennie Mayfield2, and Victoria Fraser3 And. M. luteus is an obligate aerobe (Medical Laboratories). 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. 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. They grow on simple media and their colonies are usually pigmented. My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. 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. 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. Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. M. luteus is considered an opportunistic pathogen that can be responsible for some of the nosocomial infections. The oxidase test tests to see if the microbe contains cytochrome c oxidase. 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. The colony morphology of being yellow, shiny and smooth line up perfectly with M. luteus (Public Health England). 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. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. They are considered as normal comensal of human skin and upper respiratory tract. As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. The Kocuria rhizophila strain ATCC 9341 has been used in quality control for sterility testing, as a test for the effectiveness of antibiotics and fungicides, and for doxycycline, tetracycline, and chloramphenicol susceptibility testing since 1966 under the designation Micrococcus luteus [4]. The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. The differentiation is based on the detection of oxidase enzyme. M. luteus present on the human skin can transform compounds present in sweat into compounds with an unpleasant odour. Each family of Gram-positive cocci has its own unique configuration. Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. 2002 can be divided into three biovars that are distinguished by several chemotaxonomic and biochemical traits: biovar I represented by the type strain of Micrococcus luteus; biovar II represented by strain D7 DSM 14234 CCM 4959; and biovar III . 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 nasal cavity microbiota of healthy adults. luteus is capable of surviving in environments Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. Last updated: August 10, 2022 by Sagar Aryal. A summary table is included which lists the results of the identification criteria discussed in the tutorial. 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. Your email address will not be published. They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. 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. Micrococcus luteus biochemical test result. Wikipedia also says that Micrococcus luteus is an obligate aerobe, backing up what my results show (2019). All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. This can occur due to the presence of a reduced number of proteins that can bind to penicillin. 2014). "EPR and ENDOR detection of compound I from Micrococcus lysodeikticus catalase". Catalase is an enzyme produced by the organism to neutralize the toxic effects of hydrogen peroxide in the environment of the cell. Micrococcus species are strictly aerobic Gram-positive cocci arranged in tetrads or irregular clusters, and cells range from 0.5 to 3 m in diameter. 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 . Although it's tempting to write a summary of the entire study but it . Catalase test was the first biochemical test to be done . Culture Characteristics: Colonies are shades of yellow or red. They are generally strict aerobes and can generally reduce nitrate. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . These families of bacteria produce colonies which are variable both in size, consistency, and the type of hemolysis produced on sBAP. 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). 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. CCTC (English) Klebsiella pneumoniae Micrococcus luteus Micrococcus roseus Proteus mirabilis Proteus vulgaris Pseudomonas aeruginosa Salmonella typhimurium Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Streptococcus . The large polysaccharide molecule starch contains two parts, amylose and amylopectin, these are rapidly hydrolyzed using a hydrolase called alpha-amylase to produce smaller molecules: dextrins, maltose, and glucose. The microbiome of the nose. (2019, April 4). CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. Coagulase, which is an enzyme involved in the conversion of serum fibrinogen to fibrin, causes the clotting of plasma. Micrococcus luteus biochemical test result. . If an organism is capable of fermenting the sugar glucose, then acidic byproducts are formed and the pH indicator turns yellow. So far there are two genome sequences that have been done. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. This is designated as beta ()-hemolysis. Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) In contrast to staphylococci, usually, penicillin is sensitive. Though today the immuno-compromised patients take the risk of the infection that has grown. I also had to do the thyoglycate test 3 times to get a conclusive result, further making me skeptical of how active the culture was during the physical tests during week 6, which is where almost all of the inconsistencies arose. Several uncommon strains produce raised colonies with translucent, depressed centers. The oxidase test results suggest that the microbe does not contain oxidase, despite what the metagenome binning test showed. They can be found in boiling hot springs; they can be found in frigid waters in the artic.