About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. Staphylococcus aureus Staphylococcus spp. It can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). 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. 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. 570 . See answer (1) best answer. 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. Micrococcus luteus. It is an aerobic exopolysaccharide producing bacteria that may be found on the skin, mouth and sometimes in the throat of mammals 13,14. The partial decomposition of the hemoglobin of the erythrocytes by a weaker hemolysin. Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. This is termed alpha () hemolysis and results in a greenish color surrounding the colony. Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. Wieser et al. (negative). M. luteus is considered an opportunistic pathogen that can be responsible for some of the nosocomial infections. I then repeated this three more times to further purify the isolate. 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. Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). Most of the bacterium in the gram stains were gram negative, but a significant amount, about twenty percent, showed up as gram positive. From: Infectious Diseases (Third Edition), 2010 View all Topics About this page Micrococcus spp. 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 skin infections or chronic cutaneous infections that are caused can result in pruritic eruptions of the skin in some of the areas as well as scattered papule lesions with or without central ulcerations. In contrast to staphylococci, usually, penicillin is sensitive. 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. Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. Micrococcus also not able to ferment glucose anaerobically (negative) Reply. 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. 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. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. Micrococcus Catalase Test: The catalase test facilitates the detection of the enzyme catalase in bacteria. AACC uses cookies to ensure the best website experience. This would be read K/NC. The configurations are the result of the plane of cellular division exhibited by the organism. 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 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]. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. Characteristics. (2019, March 14). Download Scientific Diagram from www.researchgate.net. A positive test leads to the development of blue or purple-blue on the disc within two minutes. 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. Therefore I used this as my location to take my environmental sample from. Micrococcus luteus biochemical test result. Micrococcus luteus in tetrads arrangement. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. Micrococcus has a substantial cell wall in which it may comprise as much as 50% of the cell mass. Micrococcus (pleural-micrococci) is free-living in the environment and also normal flora of the skin. 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. They are generally strict aerobes and can generally reduce nitrate. Enterococcus spp. In the case of a negative test, no colour change is observed. 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. They can even survive in conditions that have very little of even no oxygen, using glycolysis and alternative electron acceptors to create ATP. The gram stain of this microbe showed that it is gram positive because it stained purple. Growth can produce colonies of a certain size and consistency, as well as pigmentation. [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]. Is Micrococcus luteus indole positive? 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. 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. 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. 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. 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. I used the PATRIC software to perform a metagenome binning and to assign a taxonomy to the bacteria. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. Micrococcus luteus pigment (Yellow) GLUCOSE FERMENTATION TEST This test detects the ability of the bacterium to break down glucose to pyruvic acid. The bacterium also colonizes the mouth, mucosae, oropharynx, and upper respiratory tract. 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. Staphylococcus spp. 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. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. The colony morphology is the same as well, being round, shiny, and sort of flat (Medical Laboratories). 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. 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. They are catalase and oxidase positive whereas urease negative. 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. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. Sarah D. Perkins., Government of Canada, Public Health Agency of Canada. The micrococci produce tetrads consisting of 4 cocci clustered together. It is a gram positive, coccus shaped microbe, and contains catalase. The bubbling reaction is almost immediate and distinct in appearance. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. Although it's tempting to write a summary of the entire study but it . The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. Bacteria Collection: Micrococcus luteus Additional Information. The taxonomic assignment of this microbe was micrococcus luteus because it was the only bin that PATRIC gave. Some are based on monoclonal antibodies, and others on simple, rapid . This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. 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. The microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. [7], In 2003, it was proposed that one strain of Micrococcus luteus, ATCC 9341, be reclassified as Kocuria rhizophila. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. Micrococcus A study with 188 micrococci, identified only to the genus level, demonstrated MICs at achievable concentrations for most -lactams, aminoglycosides, glycopeptides, clindamycin and the most active drug in vitro, rifampin. 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. 2014). Bacillus megaterium fermented sugars but didn't grow in the anaerobic area of the butt. 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. Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher Though today the immuno-compromised patients take the risk of the infection that has grown. There are around nine species that are recognized in the genus. The Culture Collections represent deposits of cultures from world-wide sources. Krona chart of microbe shows bacterial classes thought to be present in the sample. It has also been isolated from foods such as milk and goats cheese. 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. The catalase test facilitates the detection of the enzyme catalase in bacteria. The differentiation is based on the detection of the oxidase enzymes. This is designated as beta ()-hemolysis. Make a tape label writing the color dot, your name, and the name of the media. 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 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. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. 2. M. luteus causes odours in humans when breaking down the components of sweat. 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. Catalase test was the first biochemical test to be done . Genus: Micrococcus. 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. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. 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. 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. It did not reduce sulphur, digest gelatin, or reduce nitrate, among some of the more interesting tests, which all disagree with M. luteus. Your email address will not be published. Micrococci can often carry plasmids that provide the organism with useful traits. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. M. luteus is an obligate aerobe (Medical Laboratories). document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. 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. Micrococci have been isolated from the human skin, animal and dairy products, and in beer. Maximum production of pigments was observed at 35C, pH 9 and at 4% (W/V) NaCl concentration. 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. These results might not be trustworthy. This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-). 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. If I were to continue researching this isolate, I would redo the API 20 E test strip with a fresh, active culture to ensure that it can reduce nitrate, and also the oxidase test to ensure that it does have cytochrome c oxidase present, which it should according to Public Health England. Micrococcus luteus is a coccus shaped gram-positive bacteria. Organisms commonly found on the body include Micrococcus, Staphylococcus, and Corynebacterium species (Dermnet New Zeland). The differentiation is based on the detection of the oxidase enzymes. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). 1995; Wieser et al. Micrococcus spp. Wikipedia also says that Micrococcus luteus is an obligate aerobe, backing up what my results show (2019). They occur in pairs, tetrads or clusters but not in chains. I grew my isolated in Tryptic Soy Broth (TSB) for a week to prepare for DNA extraction. The microbiome of the nose. // Colonies typically have a lemon-yellow pigment. 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. Figure 2. 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These residential skin organisms are also fairly ubiquitous in the environment. What are the Uses of Micrococcus Gram Stain? 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. Figure 1. . luteus is capable of surviving in environments 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. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. // 2. Exposure to these wavelengths of ultraviolet light has been correlated with an increased incidence of skin cancer, and scientists believe this pigment can be used to make a sunscreen that can protect against ultraviolet light.[9]. I would hypothesise that because we clean our shower regularly, that most of the bacterium un the shower drain would come from people in my family taking a shower, and mostly be from the skin. Abstract Micrococcus luteus IFO 3333 cells, both treated with chemical reagents and non-treated, were observed with a scanning electron microscope (SEM). Pearls of Laboratory Medicine Wikipedia contributors. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. "EPR and ENDOR detection of compound I from Micrococcus lysodeikticus catalase". all 3 tests done w/SIM deeps just add Kovac's reagent for Indole test Alcaligenes faecalis (left) - . On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. Another test I did to identify my isolate was DNA analysis, described in lab handouts 5 and 7. Enterococcus faecalis. Examples of the colony morphology associated with each family is represented on this slide. So far there are two genome sequences that have been done. 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. Coagulase production is used as a virulence factor by the organism. This microbe is coccus shaped and forms in tetrads. Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar. 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. 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. They are generally strict aerobes and these can generally reduce nitrate concentration. Micrococcus species by the Taxo A Bacitracin disk test can also be differentiated from staphylococcus species. The nasal cavity microbiota of healthy adults. Each family of Gram-positive cocci has its own unique configuration. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. I performed a Gram stain test to identify if the bacterium was gram negative or positive, as described in the Lab 4 handout. 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. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide Only the Micrococcus luteus species contained cytochrome c oxidase. 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. It had 27,372 contigs in assembly. 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. They are considered as normal comensal of human skin and upper respiratory tract. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. 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. Source publication +9 Isolation and Identification of Potential Pathogenic Bacteria in Living Carp (Cyprinus carpio Linnaeus, 1758) Sold in. A summary table is included which lists the results of the identification criteria discussed in the tutorial. 1. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. I repeated this process three more times to further purify the colony. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. Date: FEB.11.2016 I think that this culture was mostly Micrococcus luteus based on the Kaiju and metagenome binning results. 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. 1 Nevertheless they have been documented to be causative organisms in cases of bacteremia, endocarditis, ventriculitis, peritonitis, pneumonia, endophthalmitis, keratolysis and septic arthritis. In the final section of a study, it is essential to present important information, techniques, and variables. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. The tetrads of the micrococci, the grape like clusters of the staphylococci, and the pairs and/or chains of the streptococci are shown. Optimum growth temperature is 25-37 C. M. luteus has a G-C content of about 65-75 mol%. A positive test leads to the development of blue or purple-blue on the disc within two minutes. Once the culture was deemed pure enough, I inoculated a slant tube. [2] It resists antibiotic treatment by slowing of major metabolic processes and induction of unique genes[citation needed]. I kept the plate at room temperature for 7 days, and then selected a colony to purify using the pure culture streak plate method. An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal microbiota of the mammalian skin. 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. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. 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. The API 20 Strep test I used came up with no conclusive results. (2010) What are microbes? M. luteus can also cause septic shocks in immuno-compromised people. On this Wikipedia the language links are at the top of the page across from the article title. 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). 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.
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