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. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . There have been several deaths in immuno-compromised children that are caused by. The tetrads of the micrococci, the grape like clusters of the staphylococci, and the pairs and/or chains of the streptococci are shown. Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. . M. luteus is considered an opportunistic pathogen that can be responsible for some of the nosocomial infections. Some of the Micrococcus are pigmented bacteria, for example, M. roseus produces reddish colonies and M. luteus produces yellow colonies. 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. This is termed alpha () hemolysis and results in a greenish color surrounding the colony. Micrococci have been isolated from the human skin, animal and dairy products, and in beer. M. luteus is found in the soil, dust, water, and human skin flora. Catalase is an enzyme produced by the organism to neutralize the toxic effects of hydrogen peroxide in the environment of the cell. In future works with this microbe, I probably would want to purify the culture more and redo the tests. "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. The differentiation is based on the detection of the oxidase enzymes. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). 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). A number of sophisticated tools are now available for clinical diagnosis. See answer (1) best answer. 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. They are catalase positive and often oxidase positive although this reaction may be weak (see Table 23.3 ). This test had VP, HIP, ESC, PYRA, aGAL, bGUR, bGAL, PAL, LAP, ADH, RIB, ARA, MAN, SOR, LAC, TRE, INU, RAF, AMD, and GLYG tests. Staphylococci are either producers of the enzyme coagulase or non-producers. 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. I used an API Strep test to determine more of the sugars the bacteria could ferment. They are likely involved in the biodegradation of many other environmental pollutants or detoxification. Micrococcus spp. 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. What are the Diseases that are Caused By Micrococcus? Alpha-hemolytic colonies often assume a greenish hue. It has been associated with causing a variety of illnesses including septic arthritis, meningitis, endocarditis, chronic cutaneous infections in HIV-positive patients, and catheter infections. It has been isolated from human skin. CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. The conflicting results of the metagenome binning and the catalase test influenced this mistake. It is often larger thanStaphylococcus. M. luteus present on the human skin can transform compounds present in sweat into compounds with an unpleasant odour. 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. The positive catalase result lines up with M. luteus (Public Health England). It may appear in irregular clusters, groups of four or eight. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. The microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. 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 biochemical test result. 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. Gram Stain:Gram-positive cocci, arranged in clusters that differ from Staphylococcus. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. I think based on all this information, that my microbe is in fact micrococcus luteus as suggested by the PATRIC metagenome binning test, and the krona (Figure 1). I am an associate professor in the Clinical Laboratory Science program at Wayne State University. Micrococcus species occur in a wide range of environments, including dust, water, and soil. A summary table is included which lists the results of the identification criteria discussed in the tutorial. This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-). Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. Ans: Although it is a harmless saprophyte, Micrococcus luteus can act as an opportunistic pathogen. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. 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. Some Micrococcus are pigmented bacteria; for example, M. luteus produces yellow colonies and M. roseus produces redish colonies. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. In Wikipedia, The Free Encyclopedia. Most of the species produce carotenoid pigments. Wikipedia contributors. 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. Structure and PhysiologyThis bacteria is Gram-positive, spherical, and an obligate aerobe. Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. Most of the bacterium in the gram stains were gram negative, but a significant amount, about twenty percent, showed up as gram positive. 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%. It is an aerobic exopolysaccharide producing bacteria that may be found on the skin, mouth and sometimes in the throat of mammals 13,14. 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. 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 in tetrads arrangement. 2014). Micrococcus luteus was formerly known as Micrococcus lysodeikticus. The genus Micrococcus consists of Gram-positive spheres occurring in tetrads and in irregular clusters that are usually non-motile and non-spore-forming. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. Microbes are too small to be seen by the naked eye; they can survive in conditions that many would think are unlivable like the anaerobic environment in the rumen of cows, hot springs, and cold Antarctic waters (What are microbes, 2010). Therefore I used this as my location to take my environmental sample from. They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. 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. Its genome has a high G + C content. Some of the species of Micrococcus, such as M. luteus which is yellow in colour, and M. roseus which is red. (negative). 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. The antibiotic resistance test showed only minor resistance to the antibiotic Oxacillin, which is likely due to a chance inheritance in the population or complete chance because of the weak strength. It can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). They are positive for catalase and oxidase ( modified). Micrococci are usually not pathogenic. Micrococci may be distinguished from staphylococci by a modified oxidase test as shown below-. These organisms range in virulence potential from commensal organisms found on the skin and mucous membranes to pathogens associated with serious infections. In contrast to staphylococci, usually, penicillin is sensitive. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. The oxidase test was negative, as the strip did not change color at all, where it would have turned purple if it was positive. 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 . It is Gram positive, Coagulase positive, catalase and oxidase positive non-motile bacteria. This would be read A/NC. Micrococcus Public Health Agency of Canada., Public Health England. appear as agents of infection causing endocarditis. Characteristics. Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher 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. luteus is rarely found to be responsible for infections. Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). For example, M. luteus and M. lylae are 40-50% genetically different. Continuing without changing cookie settings assumes you consent to our use of cookies on this device. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. It is difficult to differentiate Kocuria from Micrococcus with conventional biochemical methods; therefore, these organisms are often reported together as Micrococcus/Kocuria species. The EMB agar showed no growth or change in color, also indicating the microbe was gram positive and a non-fermenter. Washington, DC 20001 Several uncommon strains produce raised colonies with translucent, depressed centers. Habitat Micrococcus lives in a wide range of environments such as water, soil, and the dust whereas, Staphylococcus lives in the animal body. On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. Welcome to this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. The API 20 Strep test I used came up with no conclusive results. The results of these tests provide a biochemical profile, or "fingerprint," that can be used to classify or even identify the bacterial species. Like MSA, this medium also contains the pH indicator, phenol red. This microbe is coccus shaped and forms in tetrads. What is the biochemical test for Micrococci roseus? They are positive for catalase and oxidase ( modified). Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis The coagulase test is performed by mixing a sample of organism with rabbit plasma. This microbe forms large, round colonies. Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. 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. They are catalase and oxidase positive whereas urease negative. Streptococci and enterococci are observed to produce pairs and chains of cocci of varying length. Intro: Microbes are everywhere, even in the most extreme environments and conditions. I chose to sample bacteria from inside my roommates nose. With the advent of newer molecular techniques, genus and species confirmation is even faster and easier. The nasal cavity microbiota of healthy adults. The streptococci and enterococci are categorized by expression of either beta, alpha, or gamma hemolysis on sBAP, depending on the genus/species. Unlike other actinobacteria, M. luteus expresses only one resuscitation-promoting factor that is required for the emergence from dormancy and has few other dormancy-related proteins. 2-9 In these reports micrococci were identified on the basis of their . In a temporal study by Kloos and Musselwhite (1975), it was found that micrococci usually constituted from 1 to 20% of the total aerobic bacteria isolated from the skin of the head, legs, and arms, but less than 1% of those isolated from the high bacterial density areas of the nares and axillae. It is Gram-positive cocci in tetrads, catalase-positive, coagulase-negative, arranged in clusters that differ from Staphylococcus in attacking sugars oxidatively which may appear in irregular clusters, groups of four or eight. 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 genera Staphylococcus, Micrococcus and Sarcina were recognized. Colonies typically have a lemon-yellow pigment. 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). Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. Under the microscope they are round cells. 2. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. M. luteus is part of the normal flora of the human skin.Transmission and DiseaseM. Filter paper disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride (oxidase reagent) in dimethyl sulfoxide (DMSO) are used. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. The sequencing of Micrococcus luteus was supported because of its major potential role in the case of bioremediation and its importance in the field of biotechnology. The colony morphology is the same as well, being round, shiny, and sort of flat (Medical Laboratories). Only the Micrococcus luteus species contained cytochrome c oxidase. The colonies are a pale, translucent yellow, and are shiny when looked at in the light. Isolates of the M. luteus have been found to overproduce the riboflavin virus when they are grown on toxic organic pollutants like pyridine. 1. They contain cytochromes and are resistant to lysostaphin. 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. Micrococcus luteus. Growth can produce colonies of a certain size and consistency, as well as pigmentation. Figure 2. Environmental Testing. They can be found in boiling hot springs; they can be found in frigid waters in the artic. Other Micrococcus isolates produce various useful products, such as long-chain aliphatic hydrocarbons for lubricating oils. 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. The catalase test facilitates the detection of the enzyme catalase in bacteria. 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. Only those with compromised immune systems are thought to be susceptible to an infection.DisinfectionM. Micrococcus luteus ( Scanning electron micrograph ) Micrococcus luteus is a Gram-positive bacterium of the genus Micrococcus. Wikipedia also says that Micrococcus luteus is an obligate aerobe, backing up what my results show (2019). // They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. View Biochemical Tests Lab Report.docx from BIOL 240 at University of Alaska, Anchorage. Micrococcus luteus is a Gram-positive to Gram-variable, nonmotile, tetrad-arranging, pigmented, saprotrophic coccus bacterium in the family Micrococcaceae. Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. Your email address will not be published. The PYRA, PAL, LAP, RIB, ARA, MAN, and TRE tests came up as positive. 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. Figure 1. Source publication +9 Isolation and Identification of Potential Pathogenic Bacteria in Living Carp (Cyprinus carpio Linnaeus, 1758) Sold in. 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. In Wikipedia, The Free Encyclopedia. These residential skin organisms are also fairly ubiquitous in the environment. 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 generally strict aerobes and these can generally reduce nitrate concentration. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. M. luteus has the capability to show dormancy without the formation of spores. Three bacteria, E.coli, Staphylococcus epidermis, and Micrococcus luteus, have been grown on TSI or Triple sugar iron agar medium, which is a . 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 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. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. Basics of Differentiation of Gram Positive Cocci, The Journal of Applied Laboratory Medicine, Challenges in Blood Group Alloantibody Detection, Clinical Applications of Complement Testing, Collecting Blood from Patients with Vascular Lines, Diagnosis of Syphilis Using the Reverse Algorithm, Liquid Chromatography LC Basics and Separation Techniques, Liquid Chromatography Separation Mechanisms, Optimal Reporting of Diagnostic Accuracy Studies, Pharmacogenetics for Drug Hypersensitivity Reactions, Sensitivity Specificity and Predictive Values in Diagnostic Testing, Transfusion Support in Hematopoietic Cell Transplant, Clinical Chemistry Guide to Scientific Writing, Commission on Accreditation in Clinical Chemistry. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). Reaction: To test for the presence of alpha amylase, a starch hydrolysis test can be performed. Enterococcus spp. Motility (Motile / Non-Motile) Non-Motile. They are fairly small as well, usually about a millimeter in diameter and of a normal height. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. 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. This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. Intermediates or end products of these varied metabolic activities can be detected by performing biochemical assays on a bacterial culture. Care must be taken when using a sample of organism acquired from a sBAP since erythrocytes also produce catalase. 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). It grew in circular, entire, convex and creamy yellow pigmented colonies heaving 0.5-2.5 diameter. 1995; Wieser et al. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide