micrococcus luteus biochemical tests micrococcus luteus biochemical tests

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. To identify our isolate after obtaining the pure culture, we performed many different tests. Biochemical Tests in Microbiology answer sheet - Biochemical Tests in Microbiology Pre-Laboratory - Studocu Biochemical Tests in Microbiology lab answer sheet biochemical tests in microbiology questions why do some microbes specialize to use different food source than Skip to document Ask an Expert Sign inRegister Sign inRegister Home The colonies are a pale, translucent yellow, and are shiny when looked at in the light. 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. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. 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. 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 differentiation is based on the detection of oxidase enzyme. M. lylae can be distinguished from the closely related species Micrococcus luteus by lysozyme susceptibility, genetic composition, and the type of cell-wall peptidoglycan. This technique is centered on the performance of the classic Grams stain. [3] It is a gram positive, coccus shaped microbe, and contains catalase. Biochemical Test Chart (You will not fill in every blank in this chart.) 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. There have been several deaths in immuno-compromised children that are caused by. Thus, when dealing with Micrococcus infections, it usually takes several cultures that are being grown and examined before one realizes that Micrococcus is indeed the culprit. Enterococcus spp. Table 2: Graph of the genomic reads by taxonomic level using Korona. Once the culture was deemed pure enough, I inoculated a slant tube. 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. This microbe forms large, round colonies. The gram stain of this microbe showed that it is gram positive because it stained purple. When looking back after culturing the isolate, these hypothesies were supported by several of the tests, and served as a good tool to steer me towards what the isolate may be. It has been isolated from human skin. . Micrococcaceae. 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. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . A positive test leads to the development of blue or purple-blue on the disc within two minutes. M. luteus has the capability to show dormancy without the formation of spores. 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%. Micrococcus luteus. 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. The colony morphology is the same as well, being round, shiny, and sort of flat (Medical Laboratories). Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. The results of this analysis are shown in table 1. The name of the genus is also called " Germanized " is written as Micrococcus. When performing tests for physiological traits, the results were limited. Micrococcus luteus biochemical test result. 2. Bacteria Collection: Micrococcus luteus Additional Information. 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. From: Infectious Diseases (Third Edition), 2010 View all Topics About this page The coagulase test is performed by mixing a sample of organism with rabbit plasma. Bharath. 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. Enter the email address you signed up with and we'll email you a reset link. In Wikipedia, The Free Encyclopedia. The categorization of bacteria in the clinical laboratory is based on the reactivity of the organism when stained by the Grams staining reagents. Each family of Gram-positive cocci has its own unique configuration. The metabolic pathways required for biomass production in silico were determined based on earlier models of actinobacteria. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. Culture Characteristics: Colonies are shades of yellow or red. On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. 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. Because of their diversity, there are a variety of biochemical tests that are used by laboratories to identify the Gram-positive cocci. Micrococcus., h. Shahidi Bonjar. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. 2. M. luteus is an obligate aerobe (Medical Laboratories). Another test I did to identify my isolate was DNA analysis, described in lab handouts 5 and 7. Micrococcus luteus can synthesize the iso-branched and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters. I performed a Gram stain test to identify if the bacterium was gram negative or positive, as described in the Lab 4 handout. Biochemical Pr Organism Enterococcus faecalis Lactococcus lactis Micrococcus luteus Staphylococcus 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. They are catalase-positive and usually aerobic with strictly respiratory metabolism. M. luteus causes odours in humans when breaking down the components of sweat. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). 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. 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. I hypothesized that it would be a bacteria commonly found in the nasal cavities and likely from the aforementioned phyla, so it would likely do best living in an aerobic, humid, and warm environment. The first control consisted of plates of agar-agar to test sterility. What are the Diseases that are Caused By Micrococcus? Source publication +9 Isolation and Identification of Potential Pathogenic Bacteria in Living Carp (Cyprinus carpio Linnaeus, 1758) Sold in. Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. Enterobacter aerogenes fermented the sugars but turned to the amino acids. The colony took 16 days to be purified. Where the M. luteus can cause severe skin infections and is sometimes clinically mistaken for Staphylococcus aureus. 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. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. 2. Shape (Cocci/Diplococci/Rods) Cocci. 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. 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. 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. As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. Similar to the situation in the prosthetic valve endocarditis that is caused by more aggressive and highly resistant bacteria. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. I also did a fluid thioglycolate test to determine the bacterias oxygen class. Note the bright yellow, non-diffusable colony pigment which is a defining characteristic of M. luteus. This bacterium can also be transmitted to others due to poor hand-washing practices. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). 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. I performed many tests to find out the colony morphology and physiology. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. M. luteus can also cause septic shocks in immuno-compromised people. The microbiome of the nasal cavity can also change in response to environmental factors such as geographic location, and hygiene (Rawis et al. [8], Norwegian researchers in 2013 found a M. luteus strain that synthesizes a pigment that absorbs wavelengths of light from 350 to 475 nanometers. Some Micrococcus are pigmented bacteria; for example, M. luteus produces yellow colonies and M. roseus produces redish colonies. Each genus lends itself to a separate tutorial. The staphylococci and micrococci are catalase producers while the streptococci, enterococci, and a variety of other Gram-positive cocci are catalase-negative. The data on the GC content of the DNA, chemical cell wall analysis, and a comparative analysis of the 16S rRNA sequences. This would be read A/NC. The tetrads of the micrococci, the grape like clusters of the staphylococci, and the pairs and/or chains of the streptococci are shown. M. luteus is found in the soil, dust, water, and human skin flora. I think that this culture was mostly Micrococcus luteus based on the Kaiju and metagenome binning results. // In conclusion, some of my results were inconclusive and conflicting. Isolates of the M. luteus have been found to overproduce the riboflavin virus when they are grown on toxic organic pollutants like pyridine. I grew my bacteria on an Eosin Methylene Blue (EMB) plate to see if it could ferment lactose and if it could grow with methylene blue which selects for gram negative bacteria. Examples of the colony morphology associated with each family is represented on this slide. 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. Take a clean, scratch free glass slide. It is an opportunistic pathogen, only pathogenic enough to cause disease in weakened immune systems (Medical Laboratories). 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. Figure 1. The nasal cavity microbiome primarily consists of the phyla Actinobacteria, Firmicutes and Proteobacteria (Bassis et al. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. 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You can change these settings at any time, but that may impair functionality on our websites. 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. I also grew it in a MacConkey (MAC) plate to see if it could ferment lactose and if it could grow with both crystal violet and bile salts to further confirm if it was gram negative or positive. Reaction: To test for the presence of alpha amylase, a starch hydrolysis test can be performed. 900 Seventh Street, NW Suite 400 They are catalase and oxidase positive whereas urease negative. "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. Micrococcus species are oxidase-positive, which can be used to distinguish them from other bacteria like most Staphylococcus species, which are generally oxidase-negative. Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. Whereas the M. variant has a G-C content of about 66-72mol%. Wikipedia also says that Micrococcus luteus is an obligate aerobe, backing up what my results show (2019). Many members of the genus have been reclassified into other genera. To sample, I used sterile cotton swabs and streaked them on TSA plates. 2019). An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal microbiota of the mammalian skin. 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). 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 oxygen class and the gram positiveness of the microbe also matches up with that of Micrococcus luteus. 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. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract. 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. Therefore I used this as my location to take my environmental sample from. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. I repeated this process three more times to further purify the colony. 2-9 In these reports micrococci were identified on the basis of their . 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 are catalase positive and often oxidase positive although this reaction may be weak (see Table 23.3 ). Habitat Micrococcus lives in a wide range of environments such as water, soil, and the dust whereas, Staphylococcus lives in the animal body. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. 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. Micrococcus is a spherical bacterium found on dead or decaying organic matter while Staphylococcus is a gram-positive bacterial genus that produces a bunch of grape-like bacterial clusters. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. Klebsiella pneumoniae Micrococcus luteus Micrococcus roseus Proteus mirabilis Proteus vulgaris Pseudomonas aeruginosa Salmonella typhimurium Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Streptococcus . nishinomiyaensis and Micrococcus luteus. Micrococci may be distinguished from staphylococci by a modified oxidase test as shown below-. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. Date: FEB.11.2016 So far there are two genome sequences that have been done. The optimum growth temperature is 25-37C. Micrococcus luteus | Taxonomy - PubChem Apologies, we are having some trouble retrieving data from our servers. 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. Coagulase Results: 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 can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). These can indicate that the genus of the Micrococcus is more closely related to the genus Arthrobacter than it is to other coccoid genera such as Staphylococcus and Planococcus. Environmental Testing. Staphylococcus spp. Micrococcus luteus is another gram-positive coccus that also is catalase positive. I grew my isolated in Tryptic Soy Broth (TSB) for a week to prepare for DNA extraction. The taxonomic assignment of this microbe was micrococcus luteus because it was the only bin that PATRIC gave. 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 suspect that the API 20E test strip results were negated by having used an inactive colony, as the fluid thyoglycate test failed as well. Included in the observation of the morphology of the colony is the effect that the bacterial growth has on the sheep erythrocytes in the agar medium. It is an aerobic exopolysaccharide producing bacteria that may be found on the skin, mouth and sometimes in the throat of mammals 13,14. // Other Micrococcus isolates produce various useful products, such as long-chain aliphatic hydrocarbons for lubricating oils. Micrococcus spp. They are considered as normal comensal of human skin and upper respiratory tract. The reactions of the four isolates indicated that they belonged to the species M. Zuteus (Kocur & Martinec, 1972). Intro: Microbes are everywhere, even in the most extreme environments and conditions. (2019, April 4). The microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. Although of low virulence, the germs may become more pathogenic in patients with impaired resistance, colonizing the surface of heart valves. AACC uses cookies to ensure the best website experience. As its name suggests, mannitol salt agar (MSA) contains 1% mannitol (sugar), 7.5% salt, and agar as a solidifying agent. 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). Some of the species of Micrococcus, such as M. luteus which is yellow in colour, and M. roseus which is red. Micrococcus species are oxidase-positive, where these can be used to distinguish them from some other bacteria like most of the Staphylococcus species, which are generally oxidase-negative. In the final section of a study, it is essential to present important information, techniques, and variables. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. 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. They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. After you have cultivated your organism on agar media, the next step is the microscopic examination. Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis These discrepancies could be due to human error, unpure culture, or an old agar plate. 1. appear as agents of infection causing endocarditis. Micrococcus Public Health Agency of Canada., Public Health England. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. Micrococci are usually not pathogenic. We will discuss this shortly. My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. A positive test leads to the development of blue or purple-blue on the disc within two minutes. Under the microscope they are round cells. They are generally strict aerobes and can generally reduce nitrate. 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. This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. Bacteria Detail., The Editors of Encyclopdia Britannica. (2019, March 14). This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. Although many of the members of these families have low pathogenicity and are classified as causes of opportunistic infections, those organisms most often cited as pathogens are listed on this slide. We were aiming to isolate a bacterium from this initial environmental sample by using repeated quadrant streaks to isolate single species colonies, therefore producing a pure culture that we could do a multitude of tests on which include Gram staining, genetic analysis and antibiotic testing along with many other tests. 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. Micrococcus spp. 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. Micrococcus luteus (left) Alcaligenes faecalis (middle) Reduction of nitrate to nitrite to be used as a final electron acceptor/Nitrate reductase. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. 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). They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. The differentiation of the Gram-positive cocci encompasses 4 main exercises used to compare and contrast the characteristics of each family: These four tests will, as we shall see, allow for the differentiation of the families of organisms and will guide the laboratorian to other more definitive tests for the identification of the organism. The oxidase test tests to see if the microbe contains cytochrome c oxidase. When using a fluid thyoglycollate test it resulted in the isolate being a strict aerobe, with all of the bacterium being at the top of the medium where it is oxygenic.