This presentation will focus on the laboratory tests useful for the differentiation among the families as opposed to the more complicated differentiation and identification of the organisms within the different genera. What are the Diseases that are Caused By Micrococcus? I performed many tests to find out the colony morphology and physiology. Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. When using a fluid thyoglycollate test it resulted in the isolate being a strict aerobe, with all of the bacterium being at the top of the medium where it is oxygenic. For example, Micrococcus varians is now known as Kocuria varians. 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. 2-9 In these reports micrococci were identified on the basis of their . . Bacillus megaterium fermented sugars but didn't grow in the anaerobic area of the butt. Krona chart of microbe shows bacterial classes thought to be present in the sample. // Staphylococci are either producers of the enzyme coagulase or non-producers. Retrieved 06:20, April 16, 2019, from https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=887698104, Wikipedia contributors. The microbiome of the nose. 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. I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. There are 3 biovars of this organismand they possess quite diverse chemotaxonomic features with respect to their menaquinone systems, cell-wall compositions, and Fourier transform-infrared (FT-IR) spectroscopy patterns, as well as biochemical properties. I used an API Strep test to determine more of the sugars the bacteria could ferment. Coagulase production is used as a virulence factor by the organism. This suggests that some of the Micrococcus species on the basis of ribosomal RNA analysis eventually can be re-classified into other microbial genera. This kit utilizes enzymatic reactions to release bacterial DNA from the cell. luteus is capable of surviving in environments The conflicting results of the metagenome binning and the catalase test influenced this mistake. The colony morphology is the same as well, being round, shiny, and sort of flat (Medical Laboratories). They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. Gram Staining. In Wikipedia, The Free Encyclopedia. Micrococcaceae. They are positive for catalase and oxidase ( modified). The modified oxidase reagent is prepared as tetramethyl-p-phenylenediamine in certified grade dimethyl sulfoxide. Micrococcus also not able to ferment glucose anaerobically (negative) Reply. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. Gamma or no hemolysis results in no characteristic display in the vicinity of the colony. They are positive for catalase and oxidase ( modified). From: Infectious Diseases (Third Edition), 2010 View all Topics About this page (2) Micrococcus spp. The bubbling reaction is almost immediate and distinct in appearance. It is Gram positive, Coagulase positive, catalase and oxidase positive non-motile bacteria. The microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. The nasal cavity microbiota of healthy adults. Micrococcus luteus characteristics possess unusual abilities to tolerate and to use very toxic organic molecules as carbon sources and it combines these activities with tolerance to metals. Micrococcus species by the Taxo A Bacitracin disk test can also be differentiated from staphylococcus species. Wikipedia contributors. Micrococcus Catalase Test: The catalase test facilitates the detection of the enzyme catalase in bacteria. It did not reduce sulphur, digest gelatin, or reduce nitrate, among some of the more interesting tests, which all disagree with M. luteus. After you have cultivated your organism on agar media, the next step is the microscopic examination. I then repeated this three more times to further purify the isolate. Mannitol salt agar (MSA) is a selective, differential, and indicator medium used to isolate and identify Staphylococcus aureus from the clinical specimen. 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. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. // They are catalase-positive and usually aerobic with strictly respiratory metabolism. Micrococcus species are more sensitive to the Bacitracin while the staphylococci are resistant. Only the Micrococcus luteus species contained cytochrome c oxidase. Catalase test was the first biochemical test to be done . Thank you for joining me, Karen Krisher, on this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. 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. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. 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 is a genus of bacteria that is present in the Micrococcaceae family. Micrococci may be distinguished from staphylococci by a modified oxidase test as shown below-. 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. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. It is difficult to differentiate Kocuria from Micrococcus with conventional biochemical methods; therefore, these organisms are often reported together as Micrococcus/Kocuria species. 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. 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. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. It is often larger thanStaphylococcus. Micrococcus luteus in tetrads arrangement. So far there are two genome sequences that have been done. Filter paper disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride (oxidase reagent) in dimethyl sulfoxide (DMSO) are used. 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. 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. They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. M. luteus can also cause septic shocks in immuno-compromised people. It is a gram positive, coccus shaped microbe, and contains catalase. 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. 2. When viewing the organism microscopically, the staining characteristics of the organism provide important information for identification. Kaiju webserver metagenome binning analysis chart. Micrococcus luteus. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. There are around nine species that are recognized in the genus. 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. As mentioned, the type of hemolytic reaction produced on sBAP is a major clue toward the identification of the genus. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). The species Micrococcus luteus (Schroeter 1872) Cohn 1872 (Approved Lists 1980) emend. U.S. 2022 American Association for Clinical Chemistry. 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. These residential skin organisms are also fairly ubiquitous in the environment. Obtain a glucose fermentation tube. 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. 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). M. luteus has the capability to show dormancy without the formation of spores. 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. 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. Some streptococci also possess unique cell wall carbohydrate antigens that can be identified by reactivity with specific indicator antibodies in an agglutination assay (Lancefield typing). 2002). This is termed alpha () hemolysis and results in a greenish color surrounding the colony. It grew in circular, entire, convex and creamy yellow pigmented colonies heaving 0.5-2.5 diameter. For the second control, 100 L of water sample dilutions were plated on agar-agar. The organism itself is susceptible to almost all drugs, with a few strains being resistant to nitrofurantoin, macrolides and lincomycin (Public Health Canada). Micrococcus luteus is a coccus shaped gram-positive bacteria. Whereas the M. variant has a G-C content of about 66-72mol%. The catalase test tests to see if the microbe contains catalase. Luteus is an obligate aerobe (medical. In the case of a negative test, no colour change is observed. 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. Wikipedia also says that Micrococcus luteus is an obligate aerobe, backing up what my results show (2019). Staphylococcus spp. 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%. 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. Micrococcus are aerobic, Gram-positive cocci ranging in size from 0.5 to 2.0 m in diameter. 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. Many members of the genus have been reclassified into other genera. Last updated: August 10, 2022 by Sagar Aryal. Recent reports, however, confirm that micrococci may be associated with human infections, particularly in immunosuppressed patients. The microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. 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. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. View Online Copy of Unknown ID Chart.xlsx from BIOL 250L at University of South Carolina. 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. Micrococcus luteus are Gram-positive cocci mostly arranged in tetrads and larger than Staphylococcus. Alcaligenes faecalis Bacillus megaterium . Though today the immuno-compromised patients take the risk of the infection that has grown. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). I kept the plate at room temperature for 7 days, and then selected a colony to purify using the pure culture streak plate method. M. luteus is an obligate aerobe (Medical Laboratories). Only those with compromised immune systems are thought to be susceptible to an infection.DisinfectionM. They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. The pathogen, Staphylococcus aureus, is notably coagulase-positive while most other members of the family are coagulase-negative. Shape (Cocci/Diplococci/Rods) Cocci. 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. 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. 2023 Universe84a.com | All Rights Reserved, Blog: Microbiology and infectious disease, Anti-Mullerian Hormone (AMH) Test: Introduction, Result, Unit, Normal Range, Test Method, Clinical Significance, and Keynotes, Anti -TPO Antibody: Introduction, Test Result, Unit, Normal Range, Assaying Method, and Keynotes, HPV Genes detection using Real-Time PCR: Introduction, Principle, Test Requirements, Procedure, Result Interpretation and Keynotes, Microbiology Reporting Techniques: Introduction, List of Templates, and Keynotes, Acetamide Utilization Test: Introduction, Composition, Principle, Test Requirements, Procedure, Result-Interpretation, Limitations, and Keynotes, https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/887570/UK_SMI_ID_07i4.pdf, https://en.wikipedia.org/wiki/Micrococcus_luteus, https://europepmc.org/article/med/14576986, https://www.ajicjournal.org/article/S0196-6553(13)01146-2/fulltext. This simple-to-perform test categorizes the Gram-positive cocci as either catalase producers (+) or non-producers (-). Your email address will not be published. Figure 1. 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. 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. Gram-positive organisms appear purple under the microscope as opposed to the other category of bacterial cell with its own unique cell wall, the Gram-negative rod or coccus, which stains red. 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. Required fields are marked *. I also had to do the thyoglycate test 3 times to get a conclusive result, further making me skeptical of how active the culture was during the physical tests during week 6, which is where almost all of the inconsistencies arose. 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. Where the M. luteus can cause severe skin infections and is sometimes clinically mistaken for Staphylococcus aureus. 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. M. luteus present on the human skin can transform compounds present in sweat into compounds with an unpleasant odour. A summary table is included which lists the results of the identification criteria discussed in the tutorial. 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). November 27, 2021 at 9:14 AM Hi sir, why Macrococcus and Micrococcus will not ferment . Although of low virulence, the germs may become more pathogenic in patients with impaired resistance, colonizing the surface of heart valves. I used the PATRIC software to perform a metagenome binning and to assign a taxonomy to the bacteria. It had 27,372 contigs in assembly. Micrococcus., h. Shahidi Bonjar. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. Once the culture was deemed pure enough, I inoculated a slant tube. They occur in pairs, tetrads or clusters but not in chains. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide 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. What is the biochemical test for Micrococci roseus? Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. Micrococcus species are oxidase-positive, which can be used to distinguish them from other bacteria like most Staphylococcus species, which are generally oxidase-negative. 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. They are generally strict aerobes and can generally reduce nitrate. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. Staphylococcus spp. Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. 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. The gram stain of this microbe showed that it is gram positive because it stained purple. Habitat Micrococcus lives in a wide range of environments such as water, soil, and the dust whereas, Staphylococcus lives in the animal body. 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. It can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). The genome of the Micrococcus is rich in guanine and cytosine, they typically exhibit about 65 to 75% GC-content. This technique is centered on the performance of the classic Grams stain. M. luteus played an important part in Flemings discovery of Lysozyme which is the bodys natural antibiotic. Micrococcus spp. 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.