micrococcus luteus biochemical testsfacts about witches in shakespeare's time

I also hypothesize that it will be an aerobic organism, given that I found it in a well aerated environment and it has survived until I cultured it. . Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. When looking at the genetic tests, most of the identified strains in the Korona test are Micrococcus luteus. Figure 1. M. luteus played an important part in Flemings discovery of Lysozyme which is the bodys natural antibiotic. 2. Thank you for joining me, Karen Krisher, on this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. This fits well with where I sampled my bacterium from, as a shower drain is a place where both dust and water would accumulate, along with residues of skin glands from showering. The differentiation is based on the detection of oxidase enzyme. Micrococcaceae. Production of bubbles indicates a positive reaction. This is a test for aerobic (able to use oxygen) catalase-positive bacteria such as Staphylococcus and Micrococcus. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. 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. The differentiation is based on the detection of the oxidase enzymes. 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. It shows that the sample contains bacteria from the Terrabacteria group. 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). I also did a fluid thioglycolate test to determine the bacterias oxygen class. The reactions of the four isolates indicated that they belonged to the species M. Zuteus (Kocur & Martinec, 1972). The gram stain of this microbe showed that it is gram positive because it stained purple. Annals of Allergy, Asthma and Immunology 122(1):17-24. The tetrads of the micrococci, the grape like clusters of the staphylococci, and the pairs and/or chains of the streptococci are shown. This lines up with M. luteus resistances from the tests. 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. (2010) What are microbes? Recent reports, however, confirm that micrococci may be associated with human infections, particularly in immunosuppressed patients. Intro: Microbes are everywhere, even in the most extreme environments and conditions. Next, we see an image of a mixed bacterial culture which displays the white, creamy, non- hemolytic colonies of Staphylococcus epidermidis mixed with the golden, hemolytic colonies of Staphylococcus aureus. Enterococcus faecalis. Culture Characteristics: Colonies are shades of yellow or red. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. Coagulase 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. Bacteria Detail., The Editors of Encyclopdia Britannica. 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. What is the biochemical test for Micrococci roseus? To identify our isolate after obtaining the pure culture, we performed many different tests. Like all Staphylococci, S. saprophyticus is also clustering Gram-positive cocci, nonmotile, non-spore-forming, and . M. luteus is part of the normal flora of the human skin.Transmission and DiseaseM. 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. In Wikipedia, The Free Encyclopedia. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column . I used an oxidase test strip and water to determine if cytochrome C oxidase was present, and performed a catalase test to determine if catalase was present. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). We found this to be true because the filter paper turned blue, which showed that the species has the cytochrome c oxidase enzyme. Micrococcus lylaeare Gram-positive cocci mostly arranged in tetrads. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. CCTC (English) This is termed alpha () hemolysis and results in a greenish color surrounding the colony. // Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. Structure and PhysiologyThis bacteria is Gram-positive, spherical, and an obligate aerobe. 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. Staphylococcus 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. Under the microscope they are round cells. Bacteria Collection: Micrococcus luteus Additional Information. // Some are based on monoclonal antibodies, and others on simple, rapid . The staphylococci and micrococci are catalase producers while the streptococci, enterococci, and a variety of other Gram-positive cocci are catalase-negative. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. It is a gram positive, coccus shaped microbe, and contains catalase. Most of the species produce carotenoid pigments. M. lylae can be distinguished from the closely related species Micrococcus luteus by lysozyme susceptibility, genetic composition, and the type of cell-wall peptidoglycan. Streptococcus spp. From four species that have been identify, only Bacillus subtilis and Pseudomonas diminuta that show ability to block pathogen bacteria Edwardsiella tarda. 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. 2019). Micrococcus are aerobic, Gram-positive cocci ranging in size from 0.5 to 2.0 m in diameter. There are around nine species that are recognized in the genus. Date: FEB.11.2016 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. The taxonomic assignment of this microbe was micrococcus luteus because it was the only bin that PATRIC gave. Micrococcus luteus | Taxonomy - PubChem Apologies, we are having some trouble retrieving data from our servers. Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) Only those with compromised immune systems are thought to be susceptible to an infection.DisinfectionM. The confidence on that reading is decently confident, evidenced by the blue coloring. Wieser et al. Staphylococcus spp. 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. 1995; Wieser et al. 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 ( Scanning electron micrograph ) Micrococcus luteus is a Gram-positive bacterium of the genus Micrococcus. 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). Micrococcus spp. To sample, I used sterile cotton swabs and streaked them on TSA plates. The oxygen class of the microbe, obligate aerobe, matches up with the predictions I had made about it because the bacteria was originally sourced in a nostril. Defining characteristics of Micrococcus are the ability to aerobically produce acid from glucose glycerol, aesculin hydrolysis, arginine dihydrolase, major pigment production, motility, and conversion of nitrate to nitrite. What are the Uses of Micrococcus Gram Stain? 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. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. This is designated as beta ()-hemolysis. Ans: Micrococci, like many other representatives of the Actinobacteria, can be catabolically versatile, with the ability to utilize a wide range of unusual substrates, such as herbicides, pyridine, chlorinated biphenyls, and oil. 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. 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%. // All three types of hemolytic reactions are represented on this slide. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. Micrococcus luteus, a commonly found member of the Micrococcaceae, produces round colonies which are convex (curving outward) with smooth, entire margins. (negative). (2019, April 4). 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). Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. The microbiome of the nose. The streptococci and enterococci are categorized by expression of either beta, alpha, or gamma hemolysis on sBAP, depending on the genus/species. 2. [8], Norwegian researchers in 2013 found a M. luteus strain that synthesizes a pigment that absorbs wavelengths of light from 350 to 475 nanometers. (2) Micrococcus spp. I isolated the DNA by lysing the cell and using the Power Sol DNA Isolation Kit to isolate the DNA from the other parts of the cell. The results of this analysis are shown in table 1. 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. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). In future works with this microbe, I probably would want to purify the culture more and redo the tests. They occur in pairs, tetrads or clusters but not in chains. Micrococcus luteus is generally harmless but can become an opportunistic pathogen in immunocompromised people or those with indwelling catheters. The majority of staphylococcal species produce non-pigmented colonies with smooth margins, as opposed to the important pathogen, Staphylococcus aureus, which produces the characteristic colonies seen here. Each genus lends itself to a separate tutorial. This microbe is coccus shaped and forms in tetrads. Micrococci have been isolated from the human skin, animal and dairy products, and in beer. The microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. AACC uses cookies to ensure the best website experience. 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. 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. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. Like MSA, this medium also contains the pH indicator, phenol red. Your email address will not be published. Complete lysis of the erythrocytes in the vicinity of the growing colony. (2019, March 14). Perkins1, Sarah D., Jennie Mayfield2, and Victoria Fraser3 And. They are normal inhabitants of the human body and may even be essential in keeping the balance among the various . Positive. 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. They are generally strict aerobes and these can generally reduce nitrate concentration. Required fields are marked *. CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. Shape (Cocci/Diplococci/Rods) Cocci. The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract. 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. 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. Staphylococci are either producers of the enzyme coagulase or non-producers. The catalase test also indicated that the microbe does not have catalase, despite the metagenomic binning test suggesting it. I used the PATRIC software to perform a metagenome binning and to assign a taxonomy to the bacteria. Micrococcus luteus (left) Alcaligenes faecalis (middle) Reduction of nitrate to nitrite to be used as a final electron acceptor/Nitrate reductase. 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. Book a free counselling session. 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. Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). The metabolic pathways required for biomass production in silico were determined based on earlier models of actinobacteria. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. Micrococcus species occur in a wide range of environments, including dust, water, and soil. 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 I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide Biochemical Tests in Microbiology Student Name Date 1 Lot Number(s) Record the lot numbers for each bacterial . The conflicting results of the metagenome binning and the catalase test influenced this mistake. 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. 91 988-660-2456 (Mon-Sun: 9am - 11pm IST), Want to read offline? These organisms range in virulence potential from commensal organisms found on the skin and mucous membranes to pathogens associated with serious infections. 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. Luteus is an obligate aerobe (medical. Hybridization studies indicate that species within the genus Micrococcus are not closely related, showing as little as 50% sequence similarity. You can change these settings at any time, but that may impair functionality on our websites. All pathogenic strains of S. aureus are coagulase positive whereas the nonpathogenic species (S. epidermidis for example) are coagulase negative. 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 . Krona chart of microbe shows bacterial classes thought to be present in the sample. 2002). M. luteus has one of the smallest genomes of all the bacteria. [2] It resists antibiotic treatment by slowing of major metabolic processes and induction of unique genes[citation needed]. Though today the immuno-compromised patients take the risk of the infection that has grown. 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 has been associated with causing a variety of illnesses including septic arthritis, meningitis, endocarditis, chronic cutaneous infections in HIV-positive patients, and catheter infections. The optimum growth temperature is 25-37C. // So far there are two genome sequences that have been done. 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. . Reduction of O2 produces superoxide radicals and hydrogen peroxide, or the hydroxyl radical. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. 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. 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. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. Figure 2. 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. Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. 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. Biochemical Test and Identification of Staphylococcus aureus. Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. I am an associate professor in the Clinical Laboratory Science program at Wayne State University. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). I used an API Strep test to determine more of the sugars the bacteria could ferment. 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. Wikipedia contributors. This microbe forms large, round colonies. Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. Micrococcus spp. This would be read A/NC. The identified reads only made up a total of twenty seven percent of the total reads, but the majority of those reads were for M. luteus. 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 . The catalase test facilitates the detection of the enzyme catalase in bacteria. So far there are two genome sequences that have been done. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. Examples of the colony morphology associated with each family is represented on this slide. Streptococci and enterococci are observed to produce pairs and chains of cocci of varying length. Enterococcus spp. Continuing without changing cookie settings assumes you consent to our use of cookies on this device. In the case of a negative test, no colour change is observed. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. Micrococci are microbiologically characterized as gram-positive cocci, 0.5 to 3.5 m (micrometres; 1 m = 10-6 metre) in diameter. 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. At U.S. Micro Solutions, our microbiology laboratory is a licensed, accredited source for thorough and accurate environmental testing . They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. They are likely involved in the biodegradation of many other environmental pollutants or detoxification. This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. M. luteus is found in the soil, dust, water, and human skin flora. ? The EMB agar showed no growth or change in color, also indicating the microbe was gram positive and a non-fermenter. 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. All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. Micrococcus luteus. These families of bacteria produce colonies which are variable both in size, consistency, and the type of hemolysis produced on sBAP. Only the Micrococcus luteus species contained cytochrome c oxidase. Once a presumptive identification has been made based on colony and microscopic morphology and the catalase reaction, additional tests can be performed to establish the genus and species of the organism. Alpha-hemolytic colonies often assume a greenish hue. Gram Stain:Gram-positive cocci, arranged in clusters that differ from Staphylococcus. Colony morphology, the elucidation of the characteristics of the growth of the bacterium on agar medium, in most cases trypticase soy agar containing 5% sheep blood (sBAP). all 3 tests done w/SIM deeps just add Kovac's reagent for Indole test Alcaligenes faecalis (left) - . It has also been isolated from foods such as milk and goats cheese. Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. Of those only Vancomycin was closer than 50 millimeters, being 38 millimeters. Genus: Micrococcus. An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal microbiota of the mammalian skin. 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.

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