All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. Your email address will not be published. // 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. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. A very important test in the categorization of the staphylococci is the coagulase test. Three bacteria, E.coli, Staphylococcus epidermis, and Micrococcus luteus, have been grown on TSI or Triple sugar iron agar medium, which is a . Most of the species produce carotenoid pigments. They are positive for catalase and oxidase ( modified). 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 matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). Like MSA, this medium also contains the pH indicator, phenol red. To sample, I used sterile cotton swabs and streaked them on TSA plates. Culture Characteristics: Colonies are shades of yellow or red. Each genus lends itself to a separate tutorial. As its name suggests, mannitol salt agar (MSA) contains 1% mannitol (sugar), 7.5% salt, and agar as a solidifying agent. Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. Micrococci may be distinguished from staphylococci by a modified oxidase test as shown below-. Reduction of O2 produces superoxide radicals and hydrogen peroxide, or the hydroxyl radical. The configurations are the result of the plane of cellular division exhibited by the organism. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. Download Scientific Diagram from www.researchgate.net. This is termed alpha () hemolysis and results in a greenish color surrounding the colony. Similar to the situation in the prosthetic valve endocarditis that is caused by more aggressive and highly resistant bacteria. 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. Many of the tests did line up with M. luteus though, such as the fluid thyoglycate test, which showed that it was an obligate aerobe. 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. 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. The family, Micrococcaceae, has been shrinking. All three types of hemolytic reactions are represented on this slide. Enter the email address you signed up with and we'll email you a reset link. Micrococcus species, members of the family Micrococcaceae, are usually regarded as contaminants from skin and mucous membranes. It grew in circular, entire, convex and creamy yellow pigmented colonies heaving 0.5-2.5 diameter. 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. [8], Norwegian researchers in 2013 found a M. luteus strain that synthesizes a pigment that absorbs wavelengths of light from 350 to 475 nanometers. This can occur due to the presence of a reduced number of proteins that can bind to penicillin. Alcaligenes faecalis Bacillus megaterium . CATALASE TEST: Catalases are enzymes that convert hydrogen peroxide (H2O2) into water and oxygen gas. Take a clean, scratch free glass slide. Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. They are considered as normal comensal of human skin and upper respiratory tract. 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. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. 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. To identify our isolate after obtaining the pure culture, we performed many different tests. Most of the bacterium in the gram stains were gram negative, but a significant amount, about twenty percent, showed up as gram positive. Some of the species of Micrococcus, such as M. luteus which is yellow in colour, and M. roseus which is red. 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. // This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. Washington, DC 20001 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. Beta-hemolysis is a defining characteristic of Staphylococcus aureus and a certain streptococci. 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. Staphylococcus spp. 2. Coagulase production is used as a virulence factor by the organism. The differentiation is based on the detection of oxidase enzyme. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. 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. 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. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. The genus Micrococcus consists of Gram-positive spheres occurring in tetrads and in irregular clusters that are usually non-motile and non-spore-forming. In the final section of a study, it is essential to present important information, techniques, and variables. This bacterium is often arranged in the form of circular tetrads and it forms bright yellow colonies on nutrient agar. Micrococci are usually not pathogenic. The colony took 16 days to be purified. On this Wikipedia the language links are at the top of the page across from the article title. The differentiation is based on the detection of the oxidase enzymes. I think that this culture was mostly Micrococcus luteus based on the Kaiju and metagenome binning results. Biochemical Test and Identification of Staphylococcus aureus. Micrococcus spp. Micrococcus luteus | Taxonomy - PubChem Apologies, we are having some trouble retrieving data from our servers. 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. 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. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. It had 27,372 contigs in assembly. [1] It is urease and catalase positive. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. Micrococcus luteus in tetrads arrangement. 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. 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. Oxidase (modified oxidase) test: Positive. They are also catalase-positive and often weakly oxidase-positive ( modified oxidase test positive). These families of bacteria produce colonies which are variable both in size, consistency, and the type of hemolysis produced on sBAP. The microscopic morphology of the organisms as visualized using the Grams stain can give initial clues as to the possible identification of the organism. Complete lysis of the erythrocytes in the vicinity of the growing colony. 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. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide 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). Micrococcus spp. Included in the observation of the morphology of the colony is the effect that the bacterial growth has on the sheep erythrocytes in the agar medium. I repeated this process three more times to further purify the colony. 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. Catalase test was the first biochemical test to be done . When looking at the genetic tests, most of the identified strains in the Korona test are Micrococcus luteus. Optimum growth temperature is 25-37 C. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. Only the Micrococcus luteus species contained cytochrome c oxidase. Bacteria Detail., The Editors of Encyclopdia Britannica. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . The colonies are a pale, translucent yellow, and are shiny when looked at in the light. 900 Seventh Street, NW Suite 400 Micrococcus., h. Shahidi Bonjar. Institute for Quality and Efficiency in Health Care. 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). 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 catalase test facilitates the detection of the enzyme catalase in bacteria. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. 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. The oxygen class and the gram positiveness of the microbe also matches up with that of Micrococcus luteus. I also hypothesize that it will be an aerobic organism, given that I found it in a well aerated environment and it has survived until I cultured it. Care must be taken when using a sample of organism acquired from a sBAP since erythrocytes also produce catalase. Micrococcus luteus biochemical test result. Micrococcus spp. Micrococcus luteus pigment (Yellow) GLUCOSE FERMENTATION TEST This test detects the ability of the bacterium to break down glucose to pyruvic acid. Gram Staining. 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. The gram stain of this microbe showed that it is gram positive because it stained purple. Structure and PhysiologyThis bacteria is Gram-positive, spherical, and an obligate aerobe. 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. Staphylococcus aureus Staphylococcus spp. The PYRA, PAL, LAP, RIB, ARA, MAN, and TRE tests came up as positive. Micrococci are microbiologically characterized as gram-positive cocci, 0.5 to 3.5 m (micrometres; 1 m = 10-6 metre) in diameter. Like all Staphylococci, S. saprophyticus is also clustering Gram-positive cocci, nonmotile, non-spore-forming, and . Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. Many members of the genus have been reclassified into other genera. (2019, March 14). M. luteus has a G-C content of about 65-75 mol%. Some Micrococcus are pigmented bacteria; for example, M. luteus produces yellow colonies and M. roseus produces redish colonies. 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 species by the Taxo A Bacitracin disk test can also be differentiated from staphylococcus species. 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. They can be found in boiling hot springs; they can be found in frigid waters in the artic. 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. When performing tests for physiological traits, the results were limited. Habitat Micrococcus lives in a wide range of environments such as water, soil, and the dust whereas, Staphylococcus lives in the animal body. Production of bubbles indicates a positive reaction. https://en.wikipedia.org/w/index.php?title=Micrococcus_luteus&oldid=887698104, https://en.wikipedia.org/w/index.php?title=Staphylococcus_aureus&oldid=890960280, An exploration into a pharmacists microbes, An exploration of microbes present in the blood of processed moose. It shows that the sample contains bacteria from the Terrabacteria group. The Culture Collections represent deposits of cultures from world-wide sources. They are found in soil, dust, water and air, and as part of the normal microbiota of skin. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. In Wikipedia, The Free Encyclopedia. All pathogenic strains of S. aureus are coagulase positive whereas the nonpathogenic species (S. epidermidis for example) are coagulase negative. 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. This lines up with M. luteus resistances from the tests. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. [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]. 1995; Wieser et al. 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 "EPR and ENDOR detection of compound I from Micrococcus lysodeikticus catalase". 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. There have been several deaths in immuno-compromised children that are caused by. . Figure 1. Isolates of the M. luteus have been found to overproduce the riboflavin virus when they are grown on toxic organic pollutants like pyridine. 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. Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). The microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. Although the initial placement of the Gram- positive cocci into broad categories is relatively easy, further classification of some isolates can be challenging and requires a battery of tests to arrive at a definitive identification. Bacteria Collection: Micrococcus luteus Additional Information. all 3 tests done w/SIM deeps just add Kovac's reagent for Indole test Alcaligenes faecalis (left) - . The laboratory assay is performed by mixing a sample of the bacterial colony with a drop(s) of hydrogen peroxide placed on a microscope slide. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. 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 . On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. 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. I performed a Gram stain test to identify if the bacterium was gram negative or positive, as described in the Lab 4 handout. The kaiju metagenome binning shows that the microbe sample is not completely pure (Figure 2). 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 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.

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