Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. Microbiome 2(27). 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). It has been associated with causing a variety of illnesses including septic arthritis, meningitis, endocarditis, chronic cutaneous infections in HIV-positive patients, and catheter infections. Staphylococcus spp. Some are based on monoclonal antibodies, and others on simple, rapid . They are likely involved in the biodegradation of many other environmental pollutants or detoxification. 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. I used Prokka Genome Annotation to identify which genes are present in the bacterium, Kraken Metagenomics to identify what the DNA reads correlated to on different taxonomic levels, and SPAdes Genome Assembler to determine how many contigs were produced from the reads that were produced. Krona chart of microbe shows bacterial classes thought to be present in the sample. 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 microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. Examples of the colony morphology associated with each family is represented on this slide. 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. PUGVIEW FETCH ERROR: 403 Forbidden National Center for Biotechnology Information 8600 Rockville Pike, Bethesda, MD, 20894 USA Contact Policies FOIA HHS Vulnerability Disclosure National Library of Medicine National Institutes of Health They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. They are generally strict aerobes and can generally reduce nitrate. These results might not be trustworthy. 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. I performed many tests to find out the colony morphology and physiology. 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. It is a gram positive, coccus shaped microbe, and contains catalase. Notice that beta-hemolysis results in a total lysis or clearing of the agar in the area of colony growth. Micrococci can often carry plasmids that provide the organism with useful traits. 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 . The tetrads of the micrococci, the grape like clusters of the staphylococci, and the pairs and/or chains of the streptococci are shown. 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. The differentiation is based on the detection of oxidase enzyme. Institute for Quality and Efficiency in Health Care. Enterococcus faecalis. 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. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. 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. 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). Biochemical Pr Organism Enterococcus faecalis Lactococcus lactis Micrococcus luteus Staphylococcus 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. Your email address will not be published. Micrococci are microbiologically characterized as gram-positive cocci, 0.5 to 3.5 m (micrometres; 1 m = 10-6 metre) in diameter. 2. After it was sequenced using an Illumina MiSeq at the UAF Core lab, I used the online program Base Space to analyze the reads that I got from sequencing. The data on the GC content of the DNA, chemical cell wall analysis, and a comparative analysis of the 16S rRNA sequences. A number of sophisticated tools are now available for clinical diagnosis. 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. Obtain a glucose fermentation tube. See answer (1) best answer. 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). Catalase, if present, will break down the hydrogen peroxide to water and oxygen. The coagulase test is performed by mixing a sample of organism with rabbit plasma. The differentiation is based on the detection of the oxidase enzymes. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. CCTC (English) Micrococcus species are strictly aerobic Gram-positive cocci arranged in tetrads or irregular clusters, and cells range from 0.5 to 3 m in diameter. They are catalase-positive and usually aerobic with strictly respiratory metabolism. This suggests that some of the Micrococcus species on the basis of ribosomal RNA analysis eventually can be re-classified into other microbial genera. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. 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. This microbe is coccus shaped and forms in tetrads. 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 shows that it is mostly Actinobacteria, with some firmicules ,and proteobacteria mixed in (Figure 2). Kaiju webserver metagenome binning analysis chart. Micrococcaceae. Optimum growth temperature is 25-37 C. (2019, March 14). Each genus lends itself to a separate tutorial. Micrococcus luteus This Gram positive coccus is found in tetrads, irregular clusters, and cubical packets of eight. [2] It resists antibiotic treatment by slowing of major metabolic processes and induction of unique genes[citation needed]. Then to actually isolate a bacterium, I chose a colony from the initial plate and did a quadrant streak of it to further isolate the bacterium, and then incubated it at 38 degrees Celsius for a week. Its genome has a high G + C content. Micrococcus has a substantial cell wall in which it may comprise as much as 50% of the cell mass. The oxidase test results suggest that the microbe does not contain oxidase, despite what the metagenome binning test showed. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. all 3 tests done w/SIM deeps just add Kovac's reagent for Indole test Alcaligenes faecalis (left) - . Benecky M. J.; Frew J. E.; Scowen N; Jones P, Hoffman B. M (1993). There have been several deaths in immuno-compromised children that are caused by. Washington, DC 20001 This would be read A/NC. Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis Micrococcus is the Gram-positive cocci that are about 0.5 to 3.5 micrometres in diameter and usually, these are arranged in tetrads or in irregular clusters. 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 colony took 16 days to be purified. Another test I did to identify my isolate was DNA analysis, described in lab handouts 5 and 7. 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 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. These families of bacteria produce colonies which are variable both in size, consistency, and the type of hemolysis produced on sBAP. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column . I think that this culture was mostly Micrococcus luteus based on the Kaiju and metagenome binning results. J Gen Microbiol 30(1963)409-427 . Staphylococcus aureus Staphylococcus spp. 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. It has multiple antibiotic resistance genes including dihydropteroate synthase, glycerophosphoryl diester phosphodiesterase, and SSU ribosomal proteins. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. 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Micrococcus spp. 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. Rawis M, and AK Ellis (2019). The large polysaccharide molecule starch contains two parts, amylose and amylopectin, these are rapidly hydrolyzed using a hydrolase called alpha-amylase to produce smaller molecules: dextrins, maltose, and glucose. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. 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. Intermediates or end products of these varied metabolic activities can be detected by performing biochemical assays on a bacterial culture. 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. At U.S. Micro Solutions, our microbiology laboratory is a licensed, accredited source for thorough and accurate environmental testing . Micrococcus luteuswere discovered by Sir Alexander Fleming before he discovered penicillin in 1928. Three bacteria, E.coli, Staphylococcus epidermis, and Micrococcus luteus, have been grown on TSI or Triple sugar iron agar medium, which is a . This bacterium can withstand massive doses of UV radiation and it also has the capability to degrade certain pollutants such as petrol. 1. Micrococcus luteus (left) Alcaligenes faecalis (middle) Reduction of nitrate to nitrite to be used as a final electron acceptor/Nitrate reductase. If the infection leads to severe hemodynamic alterations, thus the valve replacement may become necessary. Hemolysis is the lysis of the sheep erythrocytes within the agar by bacterial toxins (hemolysins) that are produced by the different genera of Gram-positive cocci. This can occur due to the presence of a reduced number of proteins that can bind to penicillin. 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. 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. This bacterium can also be transmitted to others due to poor hand-washing practices. 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. After observation of the colony and microscopic morphology, the production of catalase by the isolate is documented. Similar to the situation in the prosthetic valve endocarditis that is caused by more aggressive and highly resistant bacteria. Genus: Micrococcus. (2) Micrococcus spp. My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. The categorization of the Gram-positive cocci based on their possession of the enzyme catalase, a quality which can be easily assayed in the lab. . It is Gram positive, Coagulase positive, catalase and oxidase positive non-motile bacteria. The pathogen, Staphylococcus aureus, is notably coagulase-positive while most other members of the family are coagulase-negative. 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. View Biochemical Tests Lab Report.docx from BIOL 240 at University of Alaska, Anchorage. Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. // // ? The genera Staphylococcus, Micrococcus and Sarcina were recognized. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. Micrococcus luteus is a coccus shaped gram-positive bacteria. The name of the genus is also called " Germanized " is written as Micrococcus. It has been isolated from human skin. Luteus is an obligate aerobe (medical. Book a free counselling session. Required fields are marked *. This would be read K/NC. I isolated the DNA by lysing the cell and using the Power Sol DNA Isolation Kit to isolate the DNA from the other parts of the cell. 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. Enter the email address you signed up with and we'll email you a reset link. Micrococcus luteus pigment (Yellow) GLUCOSE FERMENTATION TEST This test detects the ability of the bacterium to break down glucose to pyruvic acid. The partial decomposition of the hemoglobin of the erythrocytes by a weaker hemolysin. Micrococcus spp. Some of the Micrococcus are pigmented bacteria, for example, M. roseus produces reddish colonies and M. luteus produces yellow colonies. Oxidase (modified oxidase) test: Positive. However, the most promising antibiotic regimen that is proposed for the treatment of the Micrococcus luteus seems to be a combination of amikacin, vancomycin, and rifampicin. The micrococci produce tetrads consisting of 4 cocci clustered together. The family, Micrococcaceae, has been shrinking. Recently, as an opportunistic pathogen, this organism was recognized and it has been implicated in recurrent bacteremia, septic arthritis, septic shock, endocarditis, intracranial suppuration, meningitis, and cavitating pneumonia in immunosuppressed patients. M. luteus has one of the smallest genomes of all the bacteria. The results of these tests provide a biochemical profile, or "fingerprint," that can be used to classify or even identify the bacterial species. I am an associate professor in the Clinical Laboratory Science program at Wayne State University. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. M. luteus has the capability to show dormancy without the formation of spores. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. 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.
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