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Micrococcus luteus. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. The categorization of bacteria in the clinical laboratory is based on the reactivity of the organism when stained by the Grams staining reagents. The catalase test facilitates the detection of the enzyme catalase in 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. nishinomiyaensis and Micrococcus luteus. A very important test in the categorization of the staphylococci is the coagulase test. Discussion: When looking at all of the results for my isolate, they are not all consistent with Micrococcus luteus. 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. When performing tests for physiological traits, the results were limited. M. lylae can be distinguished from the closely related species Micrococcus luteus by lysozyme susceptibility, genetic composition, and the type of cell-wall peptidoglycan. Typical colonies are often described as gray to white, moist or glistening. 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. Micrococcus luteus is a coccus shaped gram-positive bacteria. (negative). Biochemical Tests in Microbiology Student Name Date 1 Lot Number(s) Record the lot numbers for each bacterial . They are found in soil, dust, water and air, and as part of the normal microbiota of skin. View Biochemical Tests Lab Report.docx from BIOL 240 at University of Alaska, Anchorage. Know more about our courses. 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. Finally I used different antibiotic disks to check for its resistance to antibiotics, using Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, Oxacillin, and Cefazdin. Staphylococcus epidermidis contains cytochrome c which leads to turned purple after oxidation. 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. Microbiome 2(27). Enterobacter aerogenes fermented the sugars but turned to the amino acids. There are around nine species that are recognized in the genus. AACC.org 1995; Wieser et al. 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. Catalase is an enzyme produced by the organism to neutralize the toxic effects of hydrogen peroxide in the environment of the cell. Micrococcus luteus biochemical test result. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. The family, Micrococcaceae, has been shrinking. Only the Micrococcus luteus species contained cytochrome c oxidase. These results might not be trustworthy. This is a test for aerobic (able to use oxygen) catalase-positive bacteria such as Staphylococcus and Micrococcus. Required fields are marked *. Under the microscope they are round cells. This would be read as K/A. Gram Staining. All of these are described in the lab 6 handout and were kept in the incubator at 38 degrees Celsius. Enterococcus spp. Enterococcus faecalis. 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]. The agglutinability of the cells with antiserum containing anti-teichuronic acid antibody was examined. U.S. 2022 American Association for Clinical Chemistry. 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. 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. Luteus is an obligate aerobe (medical. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. M. luteus is found in the soil, dust, water, and human skin flora. 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. 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. The results of these tests provide a biochemical profile, or "fingerprint," that can be used to classify or even identify the bacterial species. 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 GC content of the DNA ranges from about 65 to 75 mol%. Micrococcus lutues are Gram positive cocci that are 0.5 to 3.5 m in diameter and arranged in tetrads or irregular clusters. Micrococcus has a substantial cell wall in which it may comprise as much as 50% of the cell mass. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. Micrococcus luteus is generally harmless but can become an opportunistic pathogen in immunocompromised people or those with indwelling catheters. Micrococcus are aerobic, Gram-positive cocci ranging in size from 0.5 to 2.0 m in diameter. It can be easily be mistaken for staphylococci, as they are very similar morphologically and physiologically (Wikipedia, Staphlyococcus Aureus). 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. Annals of Allergy, Asthma and Immunology 122(1):17-24. The Gentamicin, Cefoperazone, Vancomycin, Tobramycin, Amikacin, Trimethoprim, and Cefazdin antibiotics showed obvious susceptibility, with most of them having enormous rings of 50 to 52 millimeters while the threshold for resistance is only 15 millimeters. They are indole negative, oxidase-positive, catalase-positive, and citrate negative. [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]. 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. Biochemical Test and Identification of Staphylococcus aureus. Micrococcus luteus was formerly known as Micrococcus lysodeikticus. 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. Biochemical Pr Organism Enterococcus faecalis Lactococcus lactis Micrococcus luteus Staphylococcus The data on the GC content of the DNA, chemical cell wall analysis, and a comparative analysis of the 16S rRNA sequences. View Online Copy of Unknown ID Chart.xlsx from BIOL 250L at University of South Carolina. Growth can produce colonies of a certain size and consistency, as well as pigmentation. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. They are usually normal flora but can be opportunistic pathogens in certain immunocompromised patients. M. luteus has a G-C content of about 65-75 mol%. M. luteus is part of the normal flora of the human skin.Transmission and DiseaseM. This suggests that some of the Micrococcus species on the basis of ribosomal RNA analysis eventually can be re-classified into other microbial genera. 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. Most of the species produce carotenoid pigments. They grow on simple media and their colonies are usually pigmented. Welcome to this Pearl of Laboratory Medicine on Basics of Differentiation of Gram-positive Cocci.. This microbe forms large, round colonies. Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis . 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. Label the slide with the name of the organism; Place 15 - 20 uL of the culture in the middle of the slide [7], In 2003, it was proposed that one strain of Micrococcus luteus, ATCC 9341, be reclassified as Kocuria rhizophila. 1. Micrococcus., h. Shahidi Bonjar. The EMB agar showed no growth or change in color, also indicating the microbe was gram positive and a non-fermenter. Motility (Motile / Non-Motile) Non-Motile. If the infection leads to severe hemodynamic alterations, thus the valve replacement may become necessary. Staphylococci are either producers of the enzyme coagulase or non-producers. Micrococcus species by the Taxo A Bacitracin disk test can also be differentiated from staphylococcus species. luteus is capable of surviving in environments Gamma or no hemolysis results in no characteristic display in the vicinity of the colony. Some Micrococcus are pigmented bacteria; for example, M. luteus produces yellow colonies and M. roseus produces redish colonies. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. The oxygen class of the microbe, obligate aerobe, matches up with the predictions I had made about it because the bacteria was originally sourced in a nostril. 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. 1. 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. 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. Bharath. At U.S. Micro Solutions, our microbiology laboratory is a licensed, accredited source for thorough and accurate environmental testing . I performed many tests to find out the colony morphology and physiology. Only those with compromised immune systems are thought to be susceptible to an infection.DisinfectionM. It shows that the sample contains bacteria from the Terrabacteria group. I chose to sample bacteria from inside my roommates nose. 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. These discrepancies could be due to human error, unpure culture, or an old agar plate. The micrococci produce tetrads consisting of 4 cocci clustered together. I extracted the DNA using the PowerSoil DNA kit (manufactured by Qiagen) following manufacturer instructions. M. luteus is the majority of reads on the species level. Whereas the M. variant has a G-C content of about 66-72mol%. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. For the detection of oxidase enzyme, filter paper circular disks impregnated with tetramethyl-p-phenylenediamine dihydrochloride in dimethyl sulfoxide are used. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. 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. 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. 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. Klebsiella pneumoniae Micrococcus luteus Micrococcus roseus Proteus mirabilis Proteus vulgaris Pseudomonas aeruginosa Salmonella typhimurium Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Streptococcus . [3] If acid is produced, the pH indicator will turn yellow. 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. 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. The organism itself is susceptible to almost all drugs, with a few strains being resistant to nitrofurantoin, macrolides and lincomycin (Public Health Canada). document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. The optimum growth temperature is 25-37C. 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. In Wikipedia, The Free Encyclopedia. Micrococcus luteus | Taxonomy - PubChem Apologies, we are having some trouble retrieving data from our servers. Ans: Although it is a harmless saprophyte, Micrococcus luteus can act as an opportunistic pathogen. 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. 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. 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. For the second control, 100 L of water sample dilutions were plated on agar-agar. Abstract: Gliotoxin, an epipolythiodioxopiperazine, is a fungal metabolite that causes genomic DNA degradation preferentially in certain blood cell types including T lymphocytes and macrophages. Most of these are mesophiles; some, like Micrococcus antarcticus which is found in Antarctica, are psychrophiles. Perkins1, Sarah D., Jennie Mayfield2, and Victoria Fraser3 And. This microbe forms large, round colonies. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. 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). The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract. It is difficult to differentiate Kocuria from Micrococcus with conventional biochemical methods; therefore, these organisms are often reported together as Micrococcus/Kocuria species. It is a gram positive, coccus shaped microbe, and contains catalase. They contain cytochromes and are resistant to lysostaphin. Streptococci and enterococci are observed to produce pairs and chains of cocci of varying length. Biochemical 1- Catalase (+ve) 2- Coagulase (-ve) Coagulase production is used as a virulence factor by the organism. 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. 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%. Production of bubbles indicates a positive reaction. // The oxygen class and the gram positiveness of the microbe also matches up with that of Micrococcus luteus. Take a clean, scratch free glass slide. 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. // This kit utilizes enzymatic reactions to release bacterial DNA from the cell. 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]. The nasal cavity microbiota of healthy adults. Required fields are marked *. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. The microdase test is a rapid method to differentiate Staphylococcus (-ve) from Micrococcus +ve) by detection of the enzyme oxidase. M. luteus present on the human skin can transform compounds present in sweat into compounds with an unpleasant odour. Your email address will not be published. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. Lastly, do not underestimate the variety of different genera in the four families of the Gram- positive cocci that can, under the right circumstances, move from relatively harmless saprophytes to disease-producing opportunists. All three types of hemolytic reactions are represented on this slide. 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 have a respiratory metabolism, often producing little or no acid from carbohydrates, and are usually halotolerant, growing in 5% NaCl. These families of bacteria produce colonies which are variable both in size, consistency, and the type of hemolysis produced on sBAP. Several uncommon strains produce raised colonies with translucent, depressed centers. 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 are generally strict aerobes and can generally reduce nitrate. On this Wikipedia the language links are at the top of the page across from the article title. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. 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). This is designated as beta ()-hemolysis. Finally, when looking at the API 20E strip none of the results returned positive despite the culture being active and fresh from being streaked recently. Care must be taken when using a sample of organism acquired from a sBAP since erythrocytes also produce catalase. Micrococcus spp. all 3 tests done w/SIM deeps just add Kovac's reagent for Indole test Alcaligenes faecalis (left) - . Staphylococcus spp. 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. You can change these settings at any time, but that may impair functionality on our websites. 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. 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). The bubbling reaction is almost immediate and distinct in appearance. appear as agents of infection causing endocarditis. I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. 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. In conclusion, some of my results were inconclusive and conflicting. The microbiome of the nasal cavity can also change in response to environmental factors such as geographic location, and hygiene (Rawis et al. Hybridization studies indicate that species within the genus Micrococcus are not closely related, showing as little as 50% sequence similarity. Pearls of Laboratory Medicine Basics of Differentiation of Gram Positive Cocci, Author: Karen Krisher 2-9 In these reports micrococci were identified on the basis of their . M. luteus is considered an opportunistic pathogen that can be responsible for some of the nosocomial infections. Bassis CM, AL Tang, VB Young, and MA Pynnonen (2014). The staphylococci are characterized by the formation of clusters of many cocci. A number of sophisticated tools are now available for clinical diagnosis. Figure 1. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. Micrococcus is a genus of bacteria that is present in the Micrococcaceae family. Micrococcus spp. An interesting remaining family member, Microoccus antarcticus, was isolated from Antarctica, and is capable of growing at 4C. For example, Micrococcus varians is now known as Kocuria varians. It is an aerobic exopolysaccharide producing bacteria that may be found on the skin, mouth and sometimes in the throat of mammals 13,14. On top of that, most of the bacterium that were stained were gram negative, which conflicts with this result. From: Infectious Diseases (Third Edition), 2010 View all Topics About this page They grow in circular, entire, convex, and creamy yellow-pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37C. After you have cultivated your organism on agar media, the next step is the microscopic examination. In the case of a negative test, no colour change is observed. Positive. In order to determine physiological characteristics of the culture such as cell shape, arrangement, and whether it was gram positive or negative, which helps determine the cell wall type of the microbe, I performed a gram stain. This bacterium is often arranged in the form of circular tetrads and it forms bright yellow colonies on nutrient agar. 570 . It has also been isolated from foods such as milk and goats cheese. . My goal in this experiment was to isolate, characterize and identify a bacterial colony that arose from a sample taken from my roommates nose. Next, we see an image of a mixed bacterial culture which displays the white, creamy, non- hemolytic colonies of Staphylococcus epidermidis mixed with the golden, hemolytic colonies of Staphylococcus aureus. 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 is an opportunistic pathogen, only pathogenic enough to cause disease in weakened immune systems (Medical Laboratories). 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. Further biochemical tests to identify include the catalase and oxidase test, and typical tests of a "colorful series . Reaction: To test for the presence of alpha amylase, a starch hydrolysis test can be performed. 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. The confidence on that reading is decently confident, evidenced by the blue coloring. Another test I did to identify my isolate was DNA analysis, described in lab handouts 5 and 7. // . 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.

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