micrococcus luteus biochemical tests
Positive. The results of this analysis are shown in table 1. This would be read A/NC. 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. Micrococcus lylaeare Gram-positive cocci mostly arranged in tetrads. Reduction of O2 produces superoxide radicals and hydrogen peroxide, or the hydroxyl radical. M. luteus is an obligate aerobe (Medical Laboratories). Though today the immuno-compromised patients take the risk of the infection that has grown. For example, Micrococcus varians is now known as Kocuria varians. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. M. luteus present on the human skin can transform compounds present in sweat into compounds with an unpleasant odour. [1] It is urease and catalase positive. 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. They are normal inhabitants of the human body and may even be essential in keeping the balance among the various . Like all Staphylococci, S. saprophyticus is also clustering Gram-positive cocci, nonmotile, non-spore-forming, and . The categorization of bacteria in the clinical laboratory is based on the reactivity of the organism when stained by the Grams staining reagents. The coagulase test is performed by mixing a sample of organism with rabbit plasma. 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. There have been several deaths in immuno-compromised children that are caused by. They are found in soil, dust, water and air, and as part of the normal microbiota of skin. Although of low virulence, the germs may become more pathogenic in patients with impaired resistance, colonizing the surface of heart valves. 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. Typical colonies are often described as gray to white, moist or glistening. 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. // Micrococcus also not able to ferment glucose anaerobically (negative) Reply. The staphylococci and micrococci are catalase producers while the streptococci, enterococci, and a variety of other Gram-positive cocci are catalase-negative. They are generally strict aerobes and can generally reduce nitrate. The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. In future works with this microbe, I probably would want to purify the culture more and redo the tests. 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. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. The API 20 Strep test I used came up with no conclusive results. 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. This bacterium can also be transmitted to others due to poor hand-washing practices. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. Some of the Micrococcus are pigmented bacteria, for example, M. roseus produces reddish colonies and M. luteus produces yellow colonies. 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. 1. Each genus lends itself to a separate tutorial. Perkins1, Sarah D., Jennie Mayfield2, and Victoria Fraser3 And. ? Micrococcus spp. We found this to be true because the filter paper turned blue, which showed that the species has the cytochrome c oxidase enzyme. M. luteus is found in the soil, dust, water, and human skin flora. They can even survive in conditions that have very little of even no oxygen, using glycolysis and alternative electron acceptors to create ATP. Where the M. luteus can cause severe skin infections and is sometimes clinically mistaken for Staphylococcus aureus. 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. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. 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. Micrococcaceae. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. 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. 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. 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. All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. Required fields are marked *. 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 Annals of Allergy, Asthma and Immunology 122(1):17-24. The configurations are the result of the plane of cellular division exhibited by the organism. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. Biochemical Test and Identification of Staphylococcus aureus. 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 identification of three different biovars within the species M. luteus has the advantage that the three groups can be differentiated without nomenclatural changes having to be introduced. 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. In Wikipedia, The Free Encyclopedia. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. 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. 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. I think based on all this information, that my microbe is in fact micrococcus luteus as suggested by the PATRIC metagenome binning test, and the krona (Figure 1). 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 . Isolates of the M. luteus have been found to overproduce the riboflavin, Micrococcus Luteus Gram Stain Characteristics, Micrococci have been isolated from the human skin, animal and dairy products, and in beer. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. Organisms commonly found on the body include Micrococcus, Staphylococcus, and Corynebacterium species (Dermnet New Zeland). Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. I chose to sample bacteria from inside my roommates nose. Though today the immuno-compromised patients take the risk of the infection that has grown. Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. The catalase test tests to see if the microbe contains catalase. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. . 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. (2) Micrococcus spp. Some of the species of Micrococcus, such as M. luteus which is yellow in colour, and M. roseus which is red. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). 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. 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. Micrococcus spp. They are indole negative, oxidase-positive, catalase-positive, and citrate negative. This can occur due to the presence of a reduced number of proteins that can bind to penicillin. Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. The family, Micrococcaceae, has been shrinking. They are positive for catalase and oxidase ( modified). 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. (2019, April 4). The catalase test also indicated that the microbe does not have catalase, despite the metagenomic binning test suggesting it. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. Washington, DC 20001 Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis When performing tests for physiological traits, the results were limited. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . Micrococcus luteus ( Scanning electron micrograph ) Micrococcus luteus is a Gram-positive bacterium of the genus Micrococcus. The gram stain of this microbe showed that it is gram positive because it stained purple. . 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. The colony morphology is the same as well, being round, shiny, and sort of flat (Medical Laboratories). Alpha-hemolytic colonies often assume a greenish hue. // Your email address will not be published. The colony forms as a yellow, shiny round blob. It is often larger thanStaphylococcus. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. Biochemical Pr Organism Enterococcus faecalis Lactococcus lactis Micrococcus luteus Staphylococcus 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. View Online Copy of Unknown ID Chart.xlsx from BIOL 250L at University of South Carolina. 2. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. Colonies typically have a lemon-yellow pigment. 1. 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. So far there are two genome sequences that have been done. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. 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). Take a clean, scratch free glass slide. Only those with compromised immune systems are thought to be susceptible to an infection.DisinfectionM. 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. Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. Many of the tests did line up with M. luteus though, such as the fluid thyoglycate test, which showed that it was an obligate aerobe. If the infection leads to severe hemodynamic alterations, thus the valve replacement may become necessary. The partial decomposition of the hemoglobin of the erythrocytes by a weaker hemolysin. 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. See answer (1) best answer. J Gen Microbiol 30(1963)409-427 . Coagulase, which is an enzyme involved in the conversion of serum fibrinogen to fibrin, causes the clotting of plasma. Enter the email address you signed up with and we'll email you a reset link. 2019). Gram Stain:Gram-positive cocci, arranged in clusters that differ from Staphylococcus. Principle of Microdase (Modified 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 enzyme. Alcaligenes faecalis Bacillus megaterium . The microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. Coagulase production is used as a virulence factor by the organism. 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. For example, M. luteus and M. lylae are 40-50% genetically different. The Culture Collections represent deposits of cultures from world-wide sources. A very important test in the categorization of the staphylococci is the coagulase test. Make a tape label writing the color dot, your name, and the name of the media. Although the initial placement of the Gram- positive cocci into broad categories is relatively easy, further classification of some isolates can be challenging and requires a battery of tests to arrive at a definitive identification. This lines up with M. luteus resistances from the tests. 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. M. luteus has a G-C content of about 65-75 mol%. They are considered as normal comensal of human skin and upper respiratory tract. This suggests that some of the Micrococcus species on the basis of ribosomal RNA analysis eventually can be re-classified into other microbial genera. Welcome to this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. The colony morphology of being yellow, shiny and smooth line up perfectly with M. luteus (Public Health England). In contrast to staphylococci, usually, penicillin is sensitive. 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. Many members of the genus have been reclassified into other genera. Baird-Parker, A. c.: A classification of micrococci and staphylococci based on physiological and biochemical tests. 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). 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. 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. (2010) What are microbes? They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. Catalase, if present, will break down the hydrogen peroxide to water and oxygen. It can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). I used an API Strep test to determine more of the sugars the bacteria could ferment. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. Your email address will not be published. The differentiation is based on the detection of oxidase enzyme. I performed many tests to find out the colony morphology and physiology. Micrococcus spp. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. Micrococcus luteus is generally harmless but can become an opportunistic pathogen in immunocompromised people or those with indwelling catheters. 1995; Wieser et al. Micrococcus luteus is a coccus shaped gram-positive bacteria. 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. It has also been isolated from foods such as milk and goats cheese. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column . They are likely involved in the biodegradation of many other environmental pollutants or detoxification. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. 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. Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar. They are found in many other places in the, M. luteus has one of the smallest genomes of all the bacteria. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. 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. Bacillus megaterium fermented sugars but didn't grow in the anaerobic area of the butt. Optimum growth temperature is 25-37 C. 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. Staphylococcus aureus. M. luteus is the majority of reads on the species level. Intermediates or end products of these varied metabolic activities can be detected by performing biochemical assays on a bacterial culture. In the case of a negative test, no colour change is observed. 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. This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. 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. Growth can produce colonies of a certain size and consistency, as well as pigmentation. . From four species that have been identify, only Bacillus subtilis and Pseudomonas diminuta that show ability to block pathogen bacteria Edwardsiella tarda. These discrepancies could be due to human error, unpure culture, or an old agar plate. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . Therefore I used this as my location to take my environmental sample from. The data on the GC content of the DNA, chemical cell wall analysis, and a comparative analysis of the 16S rRNA sequences. Luteus is an obligate aerobe (medical. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. Note the bright yellow, non-diffusable colony pigment which is a defining characteristic of M. luteus. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). 2. Whereas the M. variant has a G-C content of about 66-72mol%. 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. Bacteria Collection: Micrococcus luteus Additional Information. CCTC (English) We will discuss this shortly. Maximum production of pigments was observed at 35C, pH 9 and at 4% (W/V) NaCl concentration. Filter paper disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride (oxidase reagent) in dimethyl sulfoxide (DMSO) are used. 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. 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. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. 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. 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. 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%. The colony took 16 days to be purified. On this Wikipedia the language links are at the top of the page across from the article title. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. 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. Recent reports, however, confirm that micrococci may be associated with human infections, particularly in immunosuppressed patients. [8], Norwegian researchers in 2013 found a M. luteus strain that synthesizes a pigment that absorbs wavelengths of light from 350 to 475 nanometers. The catalase test facilitates the detection of the enzyme catalase in bacteria. // 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. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. The oxidase test results suggest that the microbe does not contain oxidase, despite what the metagenome binning test showed. Micrococcus is a genus of bacteria that is present in the Micrococcaceae family. 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. 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. 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. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). The Kocuria rhizophila strain ATCC 9341 has been used in quality control for sterility testing, as a test for the effectiveness of antibiotics and fungicides, and for doxycycline, tetracycline, and chloramphenicol susceptibility testing since 1966 under the designation Micrococcus luteus [4]. 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. Micrococcus luteus in tetrads arrangement. After you have cultivated your organism on agar media, the next step is the microscopic examination. If the agar plate is held up to a light source, you can sometimes see through the agar, as is pictured on the left.