micrococcus luteus biochemical tests

. 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. Similar to the situation in the prosthetic valve endocarditis that is caused by more aggressive and highly resistant bacteria. All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. 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. In Wikipedia, The Free Encyclopedia. Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. J Gen Microbiol 30(1963)409-427 . Organisms commonly found on the body include Micrococcus, Staphylococcus, and Corynebacterium species (Dermnet New Zeland). 2002). These discrepancies could be due to human error, unpure culture, or an old agar plate. Typical colonies are often described as gray to white, moist or glistening. 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. AACC.org They are seldom motile and are non-sporing. 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. 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. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . The results of these tests provide a biochemical profile, or "fingerprint," that can be used to classify or even identify the bacterial species. 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 confidence on that reading is decently confident, evidenced by the blue coloring. 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 species Micrococcus luteus (Schroeter 1872) Cohn 1872 (Approved Lists 1980) emend. 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. Biochemical Pr Organism Enterococcus faecalis Lactococcus lactis Micrococcus luteus 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. When viewing the organism microscopically, the staining characteristics of the organism provide important information for identification. Although of low virulence, the germs may become more pathogenic in patients with impaired resistance, colonizing the surface of heart valves. 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. I also did a fluid thioglycolate test to determine the bacterias oxygen class. 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. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. It is often larger thanStaphylococcus. The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. The nasal cavity microbiome primarily consists of the phyla Actinobacteria, Firmicutes and Proteobacteria (Bassis et al. Oxidase (modified oxidase) test: Positive. Download Scientific Diagram from www.researchgate.net. 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. I chose to sample bacteria from inside my roommates nose. Make a tape label writing the color dot, your name, and the name of the media. The first control consisted of plates of agar-agar to test sterility. I think that this culture was mostly Micrococcus luteus based on the Kaiju and metagenome binning results. At U.S. Micro Solutions, our microbiology laboratory is a licensed, accredited source for thorough and accurate environmental testing . Further tests that I would do would be testing how much heat resistance it has, the density of a broth suspended sample using a dilution series, test for more antibiotic resistances, and how well it can grow in antibacterial mediums and mediums of different pH levels. 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. Last updated: August 10, 2022 by Sagar Aryal. The EMB agar showed no growth or change in color, also indicating the microbe was gram positive and a non-fermenter. [1] It is urease and catalase positive. The positive catalase result lines up with M. luteus (Public Health England). It is a gram positive, coccus shaped microbe, and contains catalase. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. View Biochemical Tests Lab Report.docx from BIOL 240 at University of Alaska, Anchorage. Where the M. luteus can cause severe skin infections and is sometimes clinically mistaken for Staphylococcus aureus. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column . 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The genus Micrococcus consists of Gram-positive spheres occurring in tetrads and in irregular clusters that are usually non-motile and non-spore-forming. Biochemical Test Chart (You will not fill in every blank in this chart.) Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. Staphylococci are either producers of the enzyme coagulase or non-producers. I used the PATRIC software to perform a metagenome binning and to assign a taxonomy to the bacteria. . Like all Staphylococci, S. saprophyticus is also clustering Gram-positive cocci, nonmotile, non-spore-forming, and . A very important test in the categorization of the staphylococci is the coagulase test. 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. Reaction: To test for the presence of alpha amylase, a starch hydrolysis test can be performed. Three bacteria, E.coli, Staphylococcus epidermis, and Micrococcus luteus, have been grown on TSI or Triple sugar iron agar medium, which is a . I kept the plate at room temperature for 7 days, and then selected a colony to purify using the pure culture streak plate method. The GC content of the DNA ranges from about 65 to 75 mol%. Complete lysis of the erythrocytes in the vicinity of the growing colony. The oxygen class and the gram positiveness of the microbe also matches up with that of Micrococcus luteus. As mentioned, the type of hemolytic reaction produced on sBAP is a major clue toward the identification of the genus. This would be read A/NC. As the microbe is gram positive this means that it has a large peptidoglycan layer and lacks a lipopolysaccharide layer. 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. 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. 91 988-660-2456 (Mon-Sun: 9am - 11pm IST), Want to read offline? 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. Bharath. Micrococcus luteuswere discovered by Sir Alexander Fleming before he discovered penicillin in 1928. In order to determine physiological characteristics of the culture such as cell shape, arrangement, and whether it was gram positive or negative, which helps determine the cell wall type of the microbe, I performed a gram stain. Some are based on monoclonal antibodies, and others on simple, rapid . They have a respiratory metabolism, often producing little or no acid from carbohydrates, and are usually halotolerant, growing in 5% NaCl. It did not reduce sulphur, digest gelatin, or reduce nitrate, among some of the more interesting tests, which all disagree with M. luteus. If the agar plate is held up to a light source, you can sometimes see through the agar, as is pictured on the left. The differentiation is based on the detection of the oxidase enzymes. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. It is reported here that gliotoxin selectively spares a unique class of haemopoietic stem cell that forms large (HPP) colonies in the presence of mixtures of MCSF and IL3. What are the Diseases that are Caused By Micrococcus? 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. Mannitol salt agar (MSA) is a selective, differential, and indicator medium used to isolate and identify Staphylococcus aureus from the clinical specimen. The categorization of bacteria in the clinical laboratory is based on the reactivity of the organism when stained by the Grams staining reagents. For example, M. luteus and M. lylae are 40-50% genetically different. Maximum production of pigments was observed at 35C, pH 9 and at 4% (W/V) NaCl concentration. You can change these settings at any time, but that may impair functionality on our websites. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. Many members of the genus have been reclassified into other genera. 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. The catalase test also indicated that the microbe does not have catalase, despite the metagenomic binning test suggesting it. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium, M. luteus played an important part in Flemings discovery of, A rare case of prosthetic valve endocarditis is caused by Micrococcus luteus, it is described and compared with the few cases that are reported in the literature, as well as the clinical features, microbiological profile, therapy, and prognosis of common prosthetic valve endocarditis. Gram Stain:Gram-positive cocci, arranged in clusters that differ from Staphylococcus. 900 Seventh Street, NW Suite 400 Micrococcus luteus is a Gram-positive to Gram-variable, nonmotile, tetrad-arranging, pigmented, saprotrophic coccus bacterium in the family Micrococcaceae. It is an opportunistic pathogen, only pathogenic enough to cause disease in weakened immune systems (Medical Laboratories). Subscribe to our e-mail newsletter to receive updates. Micrococcus luteus can synthesize the iso-branched and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters. Micrococcus spp. Micrococcus luteus is a coccus shaped gram-positive bacteria. After it was sequenced using an Illumina MiSeq at the UAF Core lab, I used the online program Base Space to analyze the reads that I got from sequencing. // The conflicting results of the metagenome binning and the catalase test influenced this mistake. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. Coagulase Results: If an organism is capable of fermenting the sugar glucose, then acidic byproducts are formed and the pH indicator turns yellow. Micrococci can often carry plasmids that provide the organism with useful traits. 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). Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. Micrococcus., h. Shahidi Bonjar. A rare case of prosthetic valve endocarditis is caused by Micrococcus luteus, it is described and compared with the few cases that are reported in the literature, as well as the clinical features, microbiological profile, therapy, and prognosis of common prosthetic valve endocarditis. This would be read as K/A. They are catalase-positive and usually aerobic with strictly respiratory metabolism. The categorization of the Gram-positive cocci based on their possession of the enzyme catalase, a quality which can be easily assayed in the lab. 1. (2019, April 4). 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 data on the GC content of the DNA, chemical cell wall analysis, and a comparative analysis of the 16S rRNA sequences. Source: Trainee Council in English, Hello, my name is Dr. Karen Krisher. They contain cytochromes and are resistant to lysostaphin. A positive test leads to the development of blue or purple-blue on the disc within two minutes. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. When performing tests for physiological traits, the results were limited. 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. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. 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. 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. These families of bacteria produce colonies which are variable both in size, consistency, and the type of hemolysis produced on sBAP. 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. The colony morphology is the same as well, being round, shiny, and sort of flat (Medical Laboratories). Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). 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. Coagulase, which is an enzyme involved in the conversion of serum fibrinogen to fibrin, causes the clotting of plasma. Some of the Micrococcus are pigmented bacteria, for example, M. roseus produces reddish colonies and M. luteus produces yellow colonies. A very important test in the categorization of the staphylococci is the coagulase test. I performed a Gram stain test to identify if the bacterium was gram negative or positive, as described in the Lab 4 handout. Whereas the M. variant has a G-C content of about 66-72mol%. Recent reports, however, confirm that micrococci may be associated with human infections, particularly in immunosuppressed patients. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. When looking at the genetic tests, most of the identified strains in the Korona test are Micrococcus luteus. 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 Take a clean, scratch free glass slide. In this presentation, we will discuss the fundamentals of the primary identification of those microorganisms that are members of four main families of Gram-positive cocci, the Micrococcaceae, the Staphylococcaceae, the Streptococcaceae, and the Enterococcaceae. Your email address will not be published. This can occur due to the presence of a reduced number of proteins that can bind to penicillin. U.S. 2022 American Association for Clinical Chemistry. Micrococci are usually not pathogenic. The bubbling reaction is almost immediate and distinct in appearance. 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. The coagulase test is performed by mixing a sample of organism with rabbit plasma. 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 . Reduction of O2 produces superoxide radicals and hydrogen peroxide, or the hydroxyl radical. Gamma or no hemolysis results in no characteristic display in the vicinity of the colony. The tests that did not agree were most likely from not having an active colony used in the experiment, such as the oxidase test being negative or the API 20 E test strip showing that the isolate did not reduce nitrate, which it does, referencing Medical Laboratories. 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. Bacteria Detail., The Editors of Encyclopdia Britannica. Micrococcus lylaeare Gram-positive cocci mostly arranged in tetrads. 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. Micrococcus as the cause of infections is easy to overlook because the infections that are caused by this bacterium are rare as well as the bacterium is a natural part of the skin's bacterial flora. They are catalase and oxidase positive whereas urease negative. For the second control, 100 L of water sample dilutions were plated on agar-agar. [7], In 2003, it was proposed that one strain of Micrococcus luteus, ATCC 9341, be reclassified as Kocuria rhizophila. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. The API 20 E test strip results revealed nothing about what the isolate uses as an electron acceptor because every result was negative, and that it likely only uses glucose as an energy and carbon source and oxygen as its electron acceptor. Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. Streptococci and enterococci are observed to produce pairs and chains of cocci of varying length. M. luteus is part of the normal flora of the human skin.Transmission and DiseaseM. 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. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. Alpha-hemolytic colonies often assume a greenish hue. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. These residential skin organisms are also fairly ubiquitous in the environment. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. (negative). 1 Nevertheless they have been documented to be causative organisms in cases of bacteremia, endocarditis, ventriculitis, peritonitis, pneumonia, endophthalmitis, keratolysis and septic arthritis. 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. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. Your email address will not be published. In the final section of a study, it is essential to present important information, techniques, and variables. The third image represents the colony morphology of many of the streptococci and enterococci. This microbe is coccus shaped and forms in tetrads. This is termed alpha () hemolysis and results in a greenish color surrounding the colony. [3] Its genome was sequenced in 2010 and is one of the smallest genomes of free-living Actinomycetota sequenced to date, comprising a single circular chromosome of 2,501,097 bp.[4]. With this almost universal range of survivable living conditions that microbes can live in (particularly bacteria), it would be reasonable to assume that there would be at least one variety living in such a nice, wet, and aerated place as my shower drain. Unlike other actinobacteria, M. luteus expresses only one resuscitation-promoting factor that is required for the emergence from dormancy and has few other dormancy-related proteins. The catalase test facilitates the detection of the enzyme catalase in bacteria. The microbiome of the nose. Luteus is an obligate aerobe (medical. Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar. M. luteus has a G-C content of about 65-75 mol%. 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 PYRA, PAL, LAP, RIB, ARA, MAN, and TRE tests came up as positive. Some of the species of Micrococcus, such as M. luteus which is yellow in colour, and M. roseus which is red. This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-).