Habitat Micrococcus lives in a wide range of environments such as water, soil, and the dust whereas, Staphylococcus lives in the animal body. Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. Micrococcus spp. Your email address will not be published. Intro: Microbes are everywhere, even in the most extreme environments and conditions. This is likely either a cause of human error, unpure cultures, or not using agar plates that are fresh enough for the test. 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. Gram Staining. Micrococcus luteuswere discovered by Sir Alexander Fleming before he discovered penicillin in 1928. Micrococcus spp. While defining the characteristics of the Micrococcus are the ability to produce acid aerobically from the aesculin hydrolysis, glucose glycerol, arginine dihydrolase, motility, major pigment production, and conversion of nitrate to nitrite. 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%. Micrococcus luteus can synthesize the iso-branched and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters. 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. Streptococcus spp. To sample, I used sterile cotton swabs and streaked them on TSA plates. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. . Wieser et al. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). You can change these settings at any time, but that may impair functionality on our websites. They are found in many other places in the environment, including dust, water, and soil. Some are based on monoclonal antibodies, and others on simple, rapid . Cells are catalase positive, oxidase positive, and exhibit strictly aerobic metabolism. The first control consisted of plates of agar-agar to test sterility. Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. Like all Staphylococci, S. saprophyticus is also clustering Gram-positive cocci, nonmotile, non-spore-forming, and . All pathogenic strains of S. aureus are coagulase positive whereas the nonpathogenic species (S. epidermidis for example) are coagulase negative. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. 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. Welcome to this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. Retrieved 06:20, April 16, 2019, from https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=887698104, Wikipedia contributors. Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. [7], In 2003, it was proposed that one strain of Micrococcus luteus, ATCC 9341, be reclassified as Kocuria rhizophila. Biochemical Test and Identification of Staphylococcus aureus. Date: FEB.11.2016 [8], Norwegian researchers in 2013 found a M. luteus strain that synthesizes a pigment that absorbs wavelengths of light from 350 to 475 nanometers. 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. M. lylae can be distinguished from the closely related species Micrococcus luteus by lysozyme susceptibility, genetic composition, and the type of cell-wall peptidoglycan. Mannitol salt agar (MSA) is a selective, differential, and indicator medium used to isolate and identify Staphylococcus aureus from the clinical specimen. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. Gram Stain:Gram-positive cocci, arranged in clusters that differ from Staphylococcus. Retrieved 22:17, April 16, 2019, from https://en.wikipedia.org/w/index.php?title=Staphylococcus_aureus&oldid=890960280, Your email address will not be published. When viewing the organism microscopically, the staining characteristics of the organism provide important information for identification. Baird-Parker, A. c.: A classification of micrococci and staphylococci based on physiological and biochemical tests. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. AACC.org In conclusion, some of my results were inconclusive and conflicting. The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. 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. M. luteus is found in the soil, dust, water, and human skin flora. This is in agreement with a study by Mohana et al., (2013) who reported that the carotenoid pigments had antioxidant properties of Micrococcus luteus was reported at IC50 of 4.5mg/ml, also the . I then repeated this three more times to further purify the isolate. With the advent of newer molecular techniques, genus and species confirmation is even faster and easier. 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. In the case of a negative test, no colour change is observed. 2002). The Gram stain, while it was gram variable, does not ideally match with the genetic test that resulted in Micrococcus luteus, which can be gram variable but is usually gram positive (Bonjar). It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. 570 . Staphylococcus aureus. Once the culture was deemed pure enough, I inoculated a slant tube. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. M. luteus played an important part in Flemings discovery of Lysozyme which is the bodys natural antibiotic. The differentiation is based on the detection of the oxidase enzymes. Micrococcus has a substantial cell wall in which it may comprise as much as 50% of the cell mass. I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. 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. 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. Gamma or no hemolysis results in no characteristic display in the vicinity of the colony. Micrococcus luteus ( Scanning electron micrograph ) Micrococcus luteus is a Gram-positive bacterium of the genus Micrococcus. Required fields are marked *. This is designated as beta ()-hemolysis. They grow in circular, entire, convex, and usually non-pigmented or cream white colonies with diameters of approximately 4 mm after 2-3 days on the plate at 37C. Micrococcus is a genus of bacteria that is present in the Micrococcaceae family. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. 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. 2. 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. The organism itself is susceptible to almost all drugs, with a few strains being resistant to nitrofurantoin, macrolides and lincomycin (Public Health Canada). This is a test for aerobic (able to use oxygen) catalase-positive bacteria such as Staphylococcus and Micrococcus. For example, M. luteus and M. lylae are 40-50% genetically different. 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. 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. All three types of hemolytic reactions are represented on this slide. // This is a test commonly used when trying to identify Gram-negative enteric bacteria, all of which are glucose fermenters but only some of which produce gas. 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. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. If an organism is capable of fermenting the sugar glucose, then acidic byproducts are formed and the pH indicator turns yellow. 2. 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). So far there are two genome sequences that have been done. 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]. Obtain a glucose fermentation tube. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. Source: Trainee Council in English, Hello, my name is Dr. Karen Krisher. Millions of microbes live both on and in the human body and can both make help us survive or make us sick, less than 1% of bacteria cause disease (What are microbes, 2010). 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. Three bacteria, E.coli, Staphylococcus epidermis, and Micrococcus luteus, have been grown on TSI or Triple sugar iron agar medium, which is a . Although it's tempting to write a summary of the entire study but it . The results of this analysis are shown in table 1. 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. Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). 1 Nevertheless they have been documented to be causative organisms in cases of bacteremia, endocarditis, ventriculitis, peritonitis, pneumonia, endophthalmitis, keratolysis and septic arthritis. 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. Rawis M, and AK Ellis (2019). Reaction: To test for the presence of alpha amylase, a starch hydrolysis test can be performed. 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. Micrococcus luteus are Gram-positive cocci mostly arranged in tetrads and larger than Staphylococcus. 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. In the case of a negative test, no colour change is observed. Complete lysis of the erythrocytes in the vicinity of the growing colony. Micrococcus Public Health Agency of Canada., Public Health England. Sarah D. Perkins., Government of Canada, Public Health Agency of Canada. They are fairly ubiquitous in the environment, and are small (0.5 to 3.5 micrometers in diameter) and non-motile. 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. The data on the GC content of the DNA, chemical cell wall analysis, and a comparative analysis of the 16S rRNA sequences. 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. Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. What is the biochemical test for Micrococci roseus? M. luteus has one of the smallest genomes of all the bacteria. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide The colonies are a pale, translucent yellow, and are shiny when looked at in the light. What are the Diseases that are Caused By Micrococcus? Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract. Micrococcus was first isolated by Alexander Fleming in 1929, as Micrococcus lysodeikticus before it was known as micrococcus luetus (Ganz et al, 2002) Micrococcus is generally thought of as harmless bacterium, but there have been rare cases of Micrococcus infections in people with compromised immune systems, as occurs with HIV patients. 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. My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. (negative). From four species that have been identify, only Bacillus subtilis and Pseudomonas diminuta that show ability to block pathogen bacteria Edwardsiella tarda. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. AACC uses cookies to ensure the best website experience. 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. 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. In future works with this microbe, I probably would want to purify the culture more and redo the tests. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. 2. 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. The positive catalase result lines up with M. luteus (Public Health England). The confidence on that reading is decently confident, evidenced by the blue coloring. Micrococcus., h. Shahidi Bonjar. 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. 1. 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. Staphylococcus spp. The colony morphology of being yellow, shiny and smooth line up perfectly with M. luteus (Public Health England). Micrococcus luteus is a coccus shaped gram-positive bacteria. Klebsiella pneumoniae Micrococcus luteus Micrococcus roseus Proteus mirabilis Proteus vulgaris Pseudomonas aeruginosa Salmonella typhimurium Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Streptococcus . They are generally strict aerobes and can generally reduce nitrate. 2014). The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. Growth can produce colonies of a certain size and consistency, as well as pigmentation. 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. The oxidase test tests to see if the microbe contains cytochrome c oxidase. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). It grew in circular, entire, convex and creamy yellow pigmented colonies heaving 0.5-2.5 diameter. The microbiome of the nose. The staphylococci are characterized by the formation of clusters of many cocci. 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. The name of the genus is also called " Germanized " is written as Micrococcus. I performed many tests to find out the colony morphology and physiology. They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. M. luteus causes odours in humans when breaking down the components of sweat. Biochemical Test Chart (You will not fill in every blank in this chart.) Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. Staphylococcus spp. Annals of Allergy, Asthma and Immunology 122(1):17-24. Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) Whereas the M. variant has a G-C content of about 66-72mol%. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). Typical colonies are often described as gray to white, moist or glistening. This bacterium is often arranged in the form of circular tetrads and it forms bright yellow colonies on nutrient agar. They are positive for catalase and oxidase ( modified). Like MSA, this medium also contains the pH indicator, phenol red. The species Micrococcus luteus (Schroeter 1872) Cohn 1872 (Approved Lists 1980) emend. Most of the bacterium in the gram stains were gram negative, but a significant amount, about twenty percent, showed up as gram positive. 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. This would be read K/NC. Micrococcus luteus, a commonly found member of the Micrococcaceae, produces round colonies which are convex (curving outward) with smooth, entire margins. The differentiation is based on the detection of the oxidase enzymes. This would be read as K/A. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. 1. This microbe is coccus shaped and forms in tetrads. It shows that it is mostly Actinobacteria, with some firmicules ,and proteobacteria mixed in (Figure 2). I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. A summary table is included which lists the results of the identification criteria discussed in the tutorial. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. 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. For example, Micrococcus varians is now known as Kocuria varians. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. 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 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. It is a gram positive, coccus shaped microbe, and contains catalase. It has also been isolated from foods such as milk and goats cheese. The majority of staphylococcal species produce non-pigmented colonies with smooth margins, as opposed to the important pathogen, Staphylococcus aureus, which produces the characteristic colonies seen here. The staphylococci and micrococci are catalase producers while the streptococci, enterococci, and a variety of other Gram-positive cocci are catalase-negative. 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 had 27,372 contigs in assembly. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. 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. It has multiple antibiotic resistance genes including dihydropteroate synthase, glycerophosphoryl diester phosphodiesterase, and SSU ribosomal proteins. Streptococcus pneumoniae and a group of streptococci referred to as viridans (green) streptococci are examples of bacteria producing alpha-hemolysis. The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. The partial decomposition of the hemoglobin of the erythrocytes by a weaker hemolysin. Because of their diversity, there are a variety of biochemical tests that are used by laboratories to identify the Gram-positive cocci. [3] 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. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. Biochemical Tests in Microbiology Student Name Date 1 Lot Number(s) Record the lot numbers for each bacterial . In contrast to staphylococci, usually, penicillin is sensitive. Of those only Vancomycin was closer than 50 millimeters, being 38 millimeters. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. For the second control, 100 L of water sample dilutions were plated on agar-agar. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. These organisms range in virulence potential from commensal organisms found on the skin and mucous membranes to pathogens associated with serious infections. 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). 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. It is Gram positive, Coagulase positive, catalase and oxidase positive non-motile bacteria. . Micrococcus luteus is coagulase negative, bacitracin susceptible, and forms bright yellow colonies on nutrient agar.