luteus is rarely found to be responsible for infections. 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. Although the Micrococcus rarely causes infections or problems in the body, with the compromised immune systems, it has been known to get certain skin infections that are caused by Micrococcus luteus. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . The optimum growth temperature is 25-37C. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) (2010) What are microbes? 2019). 1 Nevertheless they have been documented to be causative organisms in cases of bacteremia, endocarditis, ventriculitis, peritonitis, pneumonia, endophthalmitis, keratolysis and septic arthritis. As its name suggests, mannitol salt agar (MSA) contains 1% mannitol (sugar), 7.5% salt, and agar as a solidifying agent. 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). Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. Bharath. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. 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. M. luteus oxidizes carbohydrates to CO2 and water, and it does not produce acid from glucose as well as it does not make arginine dihydrolase or b-galactosidase. // The Culture Collections represent deposits of cultures from world-wide sources. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. "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 microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. I used the PATRIC software to perform a metagenome binning and to assign a taxonomy to the bacteria. Though today the immuno-compromised patients take the risk of the infection that has grown. Optimum growth temperature is 25-37 C. I think that this culture was mostly Micrococcus luteus based on the Kaiju and metagenome binning results. 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. The bubbling reaction is almost immediate and distinct in appearance. Hybridization studies indicate that species within the genus Micrococcus are not closely related, showing as little as 50% sequence similarity. Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. I then repeated this three more times to further purify the isolate. Growth can produce colonies of a certain size and consistency, as well as pigmentation. 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. Genus: Micrococcus. 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. Micrococci may be distinguished from staphylococci by a modified oxidase test as shown below-. 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). Note the bright yellow, non-diffusable colony pigment which is a defining characteristic of M. luteus. J Gen Microbiol 30(1963)409-427 . Colonies typically have a lemon-yellow pigment. Micrococcus lylaeare Gram-positive cocci mostly arranged in tetrads. Alpha-hemolytic colonies often assume a greenish hue. Micrococcus luteus. Maximum production of pigments was observed at 35C, pH 9 and at 4% (W/V) NaCl concentration. They are catalase positive and often oxidase positive although this reaction may be weak (see Table 23.3 ). Typical colonies are often described as gray to white, moist or glistening. This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. // It is difficult to differentiate Kocuria from Micrococcus with conventional biochemical methods; therefore, these organisms are often reported together as Micrococcus/Kocuria species. Structure and PhysiologyThis bacteria is Gram-positive, spherical, and an obligate aerobe. The family, Micrococcaceae, has been shrinking. The Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, and Cefazdin antibiotics showed obvious susceptibility, with most of them having enormous rings of 50 to 52 millimeters while the threshold for resistance is only 15 millimeters. I would hypothesise that because we clean our shower regularly, that most of the bacterium un the shower drain would come from people in my family taking a shower, and mostly be from the skin. The first control consisted of plates of agar-agar to test sterility. All three types of hemolytic reactions are represented on this slide. It can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). 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 is a gram positive, coccus shaped microbe, and contains catalase. 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. Micrococcus luteus characteristics possess unusual abilities to tolerate and to use very toxic organic molecules as carbon sources and it combines these activities with tolerance to metals. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. The colony morphology of being yellow, shiny and smooth line up perfectly with M. luteus (Public Health England). I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. Institute for Quality and Efficiency in Health Care. 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 . They are catalase and oxidase positive whereas urease negative. M. luteus is the majority of reads on the species level. Micrococcus luteus, a commonly found member of the Micrococcaceae, produces round colonies which are convex (curving outward) with smooth, entire margins. The coagulase test is performed by mixing a sample of organism with rabbit plasma. It did not reduce sulphur, digest gelatin, or reduce nitrate, among some of the more interesting tests, which all disagree with M. luteus. They can be found in boiling hot springs; they can be found in frigid waters in the artic. Many members of the genus have been reclassified into other genera. The staphylococci and micrococci are catalase producers while the streptococci, enterococci, and a variety of other Gram-positive cocci are catalase-negative. Micrococcus luteus ( Scanning electron micrograph ) Micrococcus luteus is a Gram-positive bacterium of the genus Micrococcus. They are catalase-positive and usually aerobic with strictly respiratory metabolism. The catalase test did return positive by bubbling, indicating that it does have the ability to break down the radical hydrogen peroxide into diatomic oxygen and hydrogen. 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. Wieser et al. On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. The species Micrococcus luteus (Schroeter 1872) Cohn 1872 (Approved Lists 1980) emend. Retrieved 06:20, April 16, 2019, from https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=887698104, Wikipedia contributors. This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. The colony took 16 days to be purified. Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. Bacteria Detail., The Editors of Encyclopdia Britannica. 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. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. Some are based on monoclonal antibodies, and others on simple, rapid . In future works with this microbe, I probably would want to purify the culture more and redo the tests. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. These residential skin organisms are also fairly ubiquitous in the environment. I performed a Gram stain test to identify if the bacterium was gram negative or positive, as described in the Lab 4 handout. Ans: Although it is a harmless saprophyte, Micrococcus luteus can act as an opportunistic pathogen. 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. I am an associate professor in the Clinical Laboratory Science program at Wayne State University. Organisms commonly found on the body include Micrococcus, Staphylococcus, and Corynebacterium species (Dermnet New Zeland). Klebsiella pneumoniae Micrococcus luteus Micrococcus roseus Proteus mirabilis Proteus vulgaris Pseudomonas aeruginosa Salmonella typhimurium Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Streptococcus . The differentiation is based on the detection of the oxidase enzymes. It has been isolated from human skin. This bacterium is often arranged in the form of circular tetrads and it forms bright yellow colonies on nutrient agar. For example, M. luteus and M. lylae are 40-50% genetically different. They are found in soil, dust, water and air, and as part of the normal microbiota of skin. The API 20 Strep test I used came up with no conclusive results. My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. It has also been isolated from foods such as milk and goats cheese. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. 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. Download Scientific Diagram from www.researchgate.net. 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. // Micrococcus luteus biochemical test result. M. luteus has a G-C content of about 65-75 mol%. The EMB plate is also selective for gram-negative bacteria which is probably why the bacteria didnt grow on it. Micrococcus is the Gram-positive cocci that are about 0.5 to 3.5 micrometres in diameter and usually, these are arranged in tetrads or in irregular clusters. Micrococcus luteus are Gram-positive cocci mostly arranged in tetrads and larger than Staphylococcus. 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. Last updated: August 10, 2022 by Sagar Aryal. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. The genus Micrococcus consists of Gram-positive spheres occurring in tetrads and in irregular clusters that are usually non-motile and non-spore-forming. The catalase test tests to see if the microbe contains catalase. They are normal inhabitants of the human body and may even be essential in keeping the balance among the various . The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. Bacillus megaterium fermented sugars but didn't grow in the anaerobic area of the butt. This would be read K/NC. Perkins1, Sarah D., Jennie Mayfield2, and Victoria Fraser3 And. Micrococcus spp. They are generally strict aerobes and can generally reduce nitrate. This bacterium can also be transmitted to others due to poor hand-washing practices. They grow on simple media and their colonies are usually pigmented. They are positive for catalase and oxidase ( modified). 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). [3] The microbiome of the nose. AACC.org In the case of a negative test, no colour change is observed. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. Alcaligenes faecalis Bacillus megaterium . https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=887698104, https://en.wikipedia.org/w/index.php?title=Staphylococcus_aureus&oldid=890960280, An exploration into a pharmacists microbes, An exploration of microbes present in the blood of processed moose. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. This would be read as K/A. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. The microbiome of the nasal cavity can also change in response to environmental factors such as geographic location, and hygiene (Rawis et al. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. Micrococcus luteus (left) Alcaligenes faecalis (middle) Reduction of nitrate to nitrite to be used as a final electron acceptor/Nitrate reductase. Kaiju webserver metagenome binning analysis chart. 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. Isolates of the M. luteus have been found to overproduce the riboflavin virus when they are grown on toxic organic pollutants like pyridine. The catalase test also indicated that the microbe does not have catalase, despite the metagenomic binning test suggesting it. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column . When looking at the genetic tests, most of the identified strains in the Korona test are Micrococcus luteus. 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. Micrococcus species by the Taxo A Bacitracin disk test can also be differentiated from staphylococcus species. Micrococci have been isolated from the human skin, animal and dairy products, and in beer. ? Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. Coagulase, which is an enzyme involved in the conversion of serum fibrinogen to fibrin, causes the clotting of plasma. Based on the place of origin of the bacterium that I isolated, I hypothesize that it will either be one of the above bacterium or a similar water loving organism, likely one that also forms a biofilm. 2- M. luteus 3- M.roseus Culture: - Strictly aerobic at 37C incubation (24 hr) - Grow on ordinary media Nutrient agar - Blood agar and on the Blood agar M. roseus (pink) M. luteus (white) M.varians (yellow) - On m annitol salt agar grow given r ise to rose or pink colonies except M. roseus. The laboratory assay is performed by mixing a sample of the bacterial colony with a drop(s) of hydrogen peroxide placed on a microscope slide. M. luteus can also cause septic shocks in immuno-compromised people. Micrococcus Catalase Test: The catalase test facilitates the detection of the enzyme catalase in bacteria. A very important test in the categorization of the staphylococci is the coagulase test. It shows that the sample contains bacteria from the Terrabacteria group. . 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. Micrococcus has a substantial cell wall in which it may comprise as much as 50% of the cell mass. 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. 2002). They are generally strict aerobes and these can generally reduce nitrate concentration. All pathogenic strains of S. aureus are coagulase positive whereas the nonpathogenic species (S. epidermidis for example) are coagulase negative. 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. They are found in many other places in the environment, including dust, water, and soil. There are around nine species that are recognized in the genus. Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. Micrococcus species occur in a wide range of environments, including dust, water, and soil. Most of the species produce carotenoid pigments. M. luteus has the capability to show dormancy without the formation of spores. Recent reports, however, confirm that micrococci may be associated with human infections, particularly in immunosuppressed patients. 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. This lines up with M. luteus resistances from the tests. 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.
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