micrococcus luteus biochemical testshow to return california license plates

There are around nine species that are recognized in the genus. 2. Table 2: Graph of the genomic reads by taxonomic level using Korona. 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. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-). The Culture Collections represent deposits of cultures from world-wide sources. The negative oxidase result conflicts with M. luteus (Public Health England). They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. 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. are gram-positive, oxidase-positive, and strictly they are aerobic cocci that are belonging to the family Micrococcaceae. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. Under the microscope they are round cells. Luteus is an obligate aerobe (medical. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. "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. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). Micrococcus luteusare Gram-positive cocci mostly arranged in tetrads and larger than Staphylococcus. Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar. It shows that it is mostly Actinobacteria, with some firmicules ,and proteobacteria mixed in (Figure 2). It has been isolated from human skin. On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. Micrococci are microbiologically characterized as gram-positive cocci, 0.5 to 3.5 m (micrometres; 1 m = 10-6 metre) in diameter. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. 2019). They are found in soil, dust, water and air, and as part of the normal microbiota of skin. Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. This would be read A/NC. appear as agents of infection causing endocarditis. The nasal cavity microbiome primarily consists of the phyla Actinobacteria, Firmicutes and Proteobacteria (Bassis et al. 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. 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. Pearls of Laboratory Medicine 2. 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. 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). The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. Micrococcus species are more sensitive to the Bacitracin while the staphylococci are resistant. They are also catalase-positive and often weakly oxidase-positive ( modified oxidase test positive). Micrococcus Public Health Agency of Canada., Public Health England. They can be found in boiling hot springs; they can be found in frigid waters in the artic. This bacterium can also be transmitted to others due to poor hand-washing practices. 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. To sample, I used sterile cotton swabs and streaked them on TSA plates. 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. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. If the agar plate is held up to a light source, you can sometimes see through the agar, as is pictured on the left. To identify our isolate after obtaining the pure culture, we performed many different tests. 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. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. 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. Abstract Micrococcus luteus IFO 3333 cells, both treated with chemical reagents and non-treated, were observed with a scanning electron microscope (SEM). View Online Copy of Unknown ID Chart.xlsx from BIOL 250L at University of South Carolina. The API 20 Strep test I used came up with no conclusive results. 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. Wieser et al. In Wikipedia, The Free Encyclopedia. A number of sophisticated tools are now available for clinical diagnosis. Micrococcus Catalase Test: The catalase test facilitates the detection of the enzyme catalase in bacteria. M. luteus has one of the smallest genomes of all the bacteria. This microbe forms large, round colonies. 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. 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 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. Structure and PhysiologyThis bacteria is Gram-positive, spherical, and an obligate aerobe. The confidence on that reading is decently confident, evidenced by the blue coloring. Note the bright yellow, non-diffusable colony pigment which is a defining characteristic of M. luteus. Only the Micrococcus luteus species contained cytochrome c oxidase. As its name suggests, mannitol salt agar (MSA) contains 1% mannitol (sugar), 7.5% salt, and agar as a solidifying agent. // Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. Micrococcus species occur in a wide range of environments, including dust, water, and soil. I performed many tests to find out the colony morphology and physiology. If acid is produced, the pH indicator will turn yellow. 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. Staphylococcus spp. Microbiome 2(27). Micrococcus luteus biochemical test result. 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. Gamma or no hemolysis results in no characteristic display in the vicinity of the colony. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. For example, Micrococcus varians is now known as Kocuria varians. Wikipedia contributors. It is a gram positive, coccus shaped microbe, and contains catalase. The differentiation is based on the detection of the oxidase enzymes. Your email address will not be published. Thank you for joining me, Karen Krisher, on this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. The tests that did not agree were most likely from not having an active colony used in the experiment, such as the oxidase test being negative or the API 20 E test strip showing that the isolate did not reduce nitrate, which it does, referencing Medical Laboratories. Micrococci have Gram-positive types of spherical cells that are ranging from about 0.5 to three micrometres in diameter and typically they appear in tetrads. As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. Care must be taken when using a sample of organism acquired from a sBAP since erythrocytes also produce catalase. 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. Results of the biochemical tests demonstrated that the M. luteus and M. varians strains could be distinguished by their actions on glucose and nitrate reduction (Table I). Micrococcus luteuswere discovered by Sir Alexander Fleming before he discovered penicillin in 1928. For example, M. luteus and M. lylae are 40-50% genetically different. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. 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 . 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. Biochemical Tests in Microbiology Student Name Date 1 Lot Number(s) Record the lot numbers for each bacterial . document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. When viewing the organism microscopically, the staining characteristics of the organism provide important information for identification. PUGVIEW FETCH ERROR: 403 Forbidden National Center for Biotechnology Information 8600 Rockville Pike, Bethesda, MD, 20894 USA Contact Policies FOIA HHS Vulnerability Disclosure National Library of Medicine National Institutes of Health Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . This microbe forms large, round colonies. 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. 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 . Obtain a glucose fermentation tube. Examples of the colony morphology associated with each family is represented on this slide. Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis This technique is centered on the performance of the classic Grams stain. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. 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. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. Staphylococci are either producers of the enzyme coagulase or non-producers. I also did a fluid thioglycolate test to determine the bacterias oxygen class. 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Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. The genome of the Micrococcus is rich in guanine and cytosine, they typically exhibit about 65 to 75% GC-content. 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). Annals of Allergy, Asthma and Immunology 122(1):17-24. Results: Regarding the gram stain test, my isolate was gram variable, I ensured that the strain was fresh so that the age of the culture was the same throughout the test. Some of the Micrococcus are pigmented bacteria, for example, M. roseus produces reddish colonies and M. luteus produces yellow colonies. A very important test in the categorization of the staphylococci is the coagulase test. 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., h. Shahidi Bonjar. All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. The metabolic pathways required for biomass production in silico were determined based on earlier models of actinobacteria. They are catalase-positive and usually aerobic with strictly respiratory metabolism. It is an opportunistic pathogen, only pathogenic enough to cause disease in weakened immune systems (Medical Laboratories). 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. Staphylococcus spp. The genus Micrococcus consists of Gram-positive spheres occurring in tetrads and in irregular clusters that are usually non-motile and non-spore-forming. Micrococcaceae. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. download full PDF here, Some of the species of Micrococcus, such as M. luteus which is yellow in colour, and M. roseus which is red. Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher Staphylococcus aureus Staphylococcus spp. Coagulase Results: The organism itself is susceptible to almost all drugs, with a few strains being resistant to nitrofurantoin, macrolides and lincomycin (Public Health Canada). The nasal cavity microbiota of healthy adults. M. lylae can be distinguished from the closely related species Micrococcus luteus by lysozyme susceptibility, genetic composition, and the type of cell-wall peptidoglycan. All three types of hemolytic reactions are represented on this slide. 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. 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. 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 GC content of the DNA ranges from about 65 to 75 mol%. 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. Millions of microbes live both on and in the human body and can both make help us survive or make us sick, less than 1% of bacteria cause disease (What are microbes, 2010). The microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. I grew my isolated in Tryptic Soy Broth (TSB) for a week to prepare for DNA extraction. Micrococcus as the cause of infections is easy to overlook because the infections that are caused by this bacterium are rare as well as the bacterium is a natural part of the skin's bacterial flora. Perkins1, Sarah D., Jennie Mayfield2, and Victoria Fraser3 And. 1995; Wieser et al. 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]. 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. 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. M. luteus is the majority of reads on the species level. Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). Enterococcus faecalis. This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. Many members of the genus have been reclassified into other genera. (2) Micrococcus spp. 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. M. luteus has a G-C content of about 65-75 mol%. The two unifying characteristics of these four families are their coccoid shape and the fact that they stain purple in the Grams method for visualizing bacteria microscopically. [2] It resists antibiotic treatment by slowing of major metabolic processes and induction of unique genes[citation needed]. "EPR and ENDOR detection of compound I from Micrococcus lysodeikticus catalase". 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. 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. Like all Staphylococci, S. saprophyticus is also clustering Gram-positive cocci, nonmotile, non-spore-forming, and . In the case of a negative test, no colour change is observed. The differentiation is based on the detection of the oxidase enzymes. The differentiation is based on the detection of oxidase enzyme. 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. . 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. In future works with this microbe, I probably would want to purify the culture more and redo the tests. 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 . (2019, March 14). Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. Micrococcus luteus is a constituent of the normal human buccal bacterial flora which forms yellowish colonies and appears as a gram-positive coccus typically arranged in tetrads. Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. This is termed alpha () hemolysis and results in a greenish color surrounding the colony. 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). Enter the email address you signed up with and we'll email you a reset link. It is often larger thanStaphylococcus. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. Micrococcus lylaeare Gram-positive cocci mostly arranged in tetrads. 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. This can occur due to the presence of a reduced number of proteins that can bind to penicillin. It had 27,372 contigs in assembly. Figure 2. Where the M. luteus can cause severe skin infections and is sometimes clinically mistaken for Staphylococcus aureus. I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. 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. The species Micrococcus luteus (Schroeter 1872) Cohn 1872 (Approved Lists 1980) emend. In conclusion, some of my results were inconclusive and conflicting. The genera Staphylococcus, Micrococcus and Sarcina were recognized. Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. Though not a spore former, the Micrococcus cells can survive for an extended period of time, both at refrigeration temperatures and in nutrient-poor conditions such as sealed in amber. Micrococcus spp. Retrieved 06:20, April 16, 2019, from https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=887698104, Wikipedia contributors. They are positive for catalase and oxidase ( modified). They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. 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. Staphylococcus saprophyticus is a Gram-positive, coagulase-negative coccus that is an important part of the group of microorganisms causing urinary tract infections (UTIs), particularly in young sexually active females. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. With this almost universal range of survivable living conditions that microbes can live in (particularly bacteria), it would be reasonable to assume that there would be at least one variety living in such a nice, wet, and aerated place as my shower drain. Gram Staining. CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. The optimum growth temperature is 25-37C. They are found in many other places in the environment, including dust, water, and soil. 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. They are catalase and oxidase positive whereas urease negative. Coagulase production is used as a virulence factor by the organism. 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. Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. 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). 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.

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