Source publication +9 Isolation and Identification of Potential Pathogenic Bacteria in Living Carp (Cyprinus carpio Linnaeus, 1758) Sold in. Klebsiella pneumoniae Micrococcus luteus Micrococcus roseus Proteus mirabilis Proteus vulgaris Pseudomonas aeruginosa Salmonella typhimurium Serratia marcescens Staphylococcus aureus Staphylococcus epidermidis Streptococcus . These results might not be trustworthy. The taxonomic assignment of this microbe was micrococcus luteus because it was the only bin that PATRIC gave. 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. To identify our isolate after obtaining the pure culture, we performed many different tests. Although it's tempting to write a summary of the entire study but it . CCTC (English) (2019, April 4). I then transferred the pure culture into a TSB slant to preserve it, keeping it at around 3 degrees Celsius in the lab refrigerator. There have been several deaths in immuno-compromised children that are caused by leukemia from the pulmonary hemorrhages because of Micrococcus. They occur in pairs, tetrads or clusters but not in chains. The third image represents the colony morphology of many of the streptococci and enterococci. It is essential for differentiating catalase-positive Micrococcaceae from catalase-negative Streptococcaceae. Gram Stain:Gram-positive cocci, arranged in clusters that differ from Staphylococcus. 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This can occur due to the presence of a reduced number of proteins that can bind to penicillin. M. luteus is part of the normal flora of the human skin.Transmission and DiseaseM. In contrast to staphylococci, usually, penicillin is sensitive. M. luteus has a G-C content of about 65-75 mol%. Coagulase, which is an enzyme involved in the conversion of serum fibrinogen to fibrin, causes the clotting of plasma. Cultures:- Nutrient broth cultures of Micrococcus luteus and Proteus vulgaris (young cultures) Procedure: Wet Mount Preparation . 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. Although of low virulence, the germs may become more pathogenic in patients with impaired resistance, colonizing the surface of heart valves. Staphylococcus aureus Staphylococcus spp. Motility (Motile / Non-Motile) Non-Motile. Biochemical Test Chart (You will not fill in every blank in this chart.) Unlike other actinobacteria, M. luteus expresses only one resuscitation-promoting factor that is required for the emergence from dormancy and has few other dormancy-related proteins. The API test strips lack of results suggests that the I used the wrong test strip, I probably needed to use the Staph test instead of the Strep test, because the Strep test is for when Catalase is absent, but there could have been catalase present. 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 catalase test facilitates the detection of the enzyme catalase in bacteria. 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. Bacteria Detail., The Editors of Encyclopdia Britannica. Environmental Testing. The differentiation is based on the detection of oxidase enzyme. It has survived for at least 34,000 to 170,000 years, as assessed by 16S rRNA analysis, and possibly much longer. The mammalian skin that has M. luteus strain is also highly sensitive to the beta-lactam antibiotics. Colony pigmentation varies considerably but is usually different shades of yellow or cream-white. Staphylococcus aureus. DNA release from the cell is coupled with adsorption of DNA onto a silica spin-column . 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. 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. I kept the plate at room temperature for 7 days, and then selected a colony to purify using the pure culture streak plate method. Several uncommon strains produce raised colonies with translucent, depressed centers. It is mostly Actinobacteria, but some Proteobacteria and Firmicules are in the sample as well. 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. 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. Micrococcus luteus ( Scanning electron micrograph ) Micrococcus luteus is a Gram-positive bacterium of the genus Micrococcus. This technique is centered on the performance of the classic Grams stain. Another test I did to identify my isolate was DNA analysis, described in lab handouts 5 and 7. 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. Wieser et al. M. luteus is the majority of reads on the species level. Growth or weak growth is observed at 45C, at pH 10, and in the presence of 10% sodium chloride (NaCl). 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. When viewing the organism microscopically, the staining characteristics of the organism provide important information for identification. Micrococcus luteus is another gram-positive coccus that also is catalase positive. 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. 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. Staphylococci were distinguished by their ability to form acid from glucose anaerobically and sarcinas by the formation of cubical packets. 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. 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. 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. Micrococcus lylaeare Gram-positive cocci mostly arranged in tetrads. The staphylococci are characterized by the formation of clusters of many cocci. (2010) What are microbes? November 27, 2021 at 9:14 AM Hi sir, why Macrococcus and Micrococcus will not ferment . This would be read as K/A. The Micrococcus strains isolated from various infections were most probably misclassified as staphylococci. See answer (1) best answer. In the 1920s, Alexander Fleming described lysozyme as a bactericidal factor of human and other animal tissues and secretions.1 He also discovered and named a yellow bacterium, Micrococcus lysodeikticus (now M luteus), that was highly susceptible to lysozyme-mediated killing.After exploring the susceptibility of other bacteria to lysozyme, Fleming proposed that M luteus and certain other . The sample was then sequenced using the Illumina MiSeq technology in UAFs DNA Core Lab. Micrococcus, genus of spherical bacteria in the family Micrococcaceae that is widely disseminated in nature. I used the Kirby-Bauer, or disk diffusion test to test for this, described in lab handout 9. This matches up with the PATRIC metagenome binning which also showed some impurities (Figure 1). Micrococci can grow well in environments with little water or high salt concentrations, including sportswear that is made with synthetic fabrics. // Micrococci can often carry plasmids that provide the organism with useful traits. 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. They are found in many other places in the, M. luteus has one of the smallest genomes of all the bacteria. We found this to be true because the filter paper turned blue, which showed that the species has the cytochrome c oxidase enzyme. Maximum production of pigments was observed at 35C, pH 9 and at 4% (W/V) NaCl concentration. Microbiome 2(27). 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. 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 (pleural-micrococci) is free-living in the environment and also normal flora of the skin. This would be read A/NC. Shape (Cocci/Diplococci/Rods) Cocci. 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. Bacteria Collection: Micrococcus luteus Additional Information. Isolates of the M. luteus have been found to overproduce the riboflavin virus when they are grown on toxic organic pollutants like pyridine. Required fields are marked *. These include the slide or drop catalase test, the tube method, the semiquantitative catalase for the identification of Mycobacterium tuberculosis. Obtain a glucose fermentation tube. They also produce yellow or pink colonies when they are grown on the mannitol salt agar, micrococcus flavus, M. terreus, and M. yunnanensis. 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]. As mentioned, the type of hemolytic reaction produced on sBAP is a major clue toward the identification of the genus. Micrococcus luteus in tetrads arrangement. Micrococcus luteus can synthesize the iso-branched and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters. The microbiome of the nasal cavity can also change in response to environmental factors such as geographic location, and hygiene (Rawis et al. They are also catalase-positive and often weakly oxidase-positive ( modified oxidase test positive). Micrococcus luteus is generally harmless but can become an opportunistic pathogen in immunocompromised people or those with indwelling catheters. Evaluation of Antibacterial Properties of Iranian Medicinal-Plants against Micrococcus Luteus, Serratia Marcescens, Klebsiella Pneumoniae and Bordetella Bronchoseptica.. Enterococcus spp. Following incubation at 37oC, if the plasma forms a clot, the organism is coagulase positive. Micrococcus spp. They are generally strict aerobes and can generally reduce nitrate. For example, Micrococcus varians is now known as Kocuria varians. document.getElementById( "ak_js_1" ).setAttribute( "value", ( new Date() ).getTime() ); Built with Enlightenment Theme and WordPress. 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. M. luteus can also cause septic shocks in immuno-compromised people. 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. Though today the immuno-compromised patients take the risk of the infection that has grown. They are fairly small as well, usually about a millimeter in diameter and of a normal height. 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 M. luteus genome encodes about four sigma factors and fourteen response regulators, a finding indicative of the adaptation to a rather strict ecological niche. They are likely involved in the biodegradation of many other environmental pollutants or detoxification. 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 configurations are the result of the plane of cellular division exhibited by the organism. 2002). Gamma or no hemolysis results in no characteristic display in the vicinity of the colony. Habitat Micrococcus lives in a wide range of environments such as water, soil, and the dust whereas, Staphylococcus lives in the animal body. Micrococcus luteus uses the amino acids and does not grow in the butt of the slant. With the advent of newer molecular techniques, genus and species confirmation is even faster and easier. Whereas the M. variant has a G-C content of about 66-72mol%. 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. We will discuss this shortly. Take a clean, scratch free glass slide. It did not reduce sulphur, digest gelatin, or reduce nitrate, among some of the more interesting tests, which all disagree with M. luteus. Note the bright yellow, non-diffusable colony pigment which is a defining characteristic of M. luteus. M. luteus is found in the soil, dust, water, and human skin flora. Rawis M, and AK Ellis (2019). Source: Trainee Council in English, Hello, my name is Dr. Karen Krisher. From four species that have been identify, only Bacillus subtilis and Pseudomonas diminuta that show ability to block pathogen bacteria Edwardsiella tarda. Micrococcus varians Micrococcus luteus Staphylococcus saprophyticus Staphylococcus epidermidis Streptococcus pneumoniae Streptococcus mitis The oxidase test results suggest that the microbe does not contain oxidase, despite what the metagenome binning test showed. It shows that the sample contains bacteria from the Terrabacteria group. Those are extremely toxic because they are powerful oxidizing agents and destroy cellular constituents very rapidly. M. luteus played an important part in Flemings discovery of Lysozyme which is the bodys natural antibiotic. Micrococcus luteus biochemical test result. 900 Seventh Street, NW Suite 400 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. They are positive for catalase and oxidase ( modified). Micrococcus luteus pigment (Yellow) GLUCOSE FERMENTATION TEST This test detects the ability of the bacterium to break down glucose to pyruvic acid. This lines up with M. luteus resistances from the tests. 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. 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. So far there are two genome sequences that have been done. For example, M. luteus and M. lylae are 40-50% genetically different. When a drop of peroxide is placed on catalase-producing bacteria, bubbles appear when the oxygen gas is formed. Microbes are too small to be seen by the naked eye; they can survive in conditions that many would think are unlivable like the anaerobic environment in the rumen of cows, hot springs, and cold Antarctic waters (What are microbes, 2010). Similar to the situation in the prosthetic valve endocarditis that is caused by more aggressive and highly resistant bacteria. Most of the bacterium in the gram stains were gram negative, but a significant amount, about twenty percent, showed up as gram positive. Micrococci have been isolated from the human skin, animal and dairy products, and in beer. They are positive for catalase and oxidase ( modified). A positive test leads to the development of blue or purple-blue on the disc within two minutes. The typical microscopic morphology of the Gram-positive cocci when using the Grams stain is represented in these three images. 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. It has been isolated from human skin. About half of the Micrococcus luteus gram stain was found to carry plasmids of about one to 100MDa in size. 1. Sarah D. Perkins., Government of Canada, Public Health Agency of Canada. Reaction: To test for the presence of alpha amylase, a starch hydrolysis test can be performed. After you have cultivated your organism on agar media, the next step is the microscopic examination. Hybridization studies show that there is no close genetic relationship between the species of Micrococcus bacteria. For the second control, 100 L of water sample dilutions were plated on agar-agar. It has been isolated from human skin. Micrococcus luteus are Gram-positive cocci mostly arranged in tetrads and larger than Staphylococcus. An obligate aerobe, M. luteus is found in soil, dust, water and air, and as part of the normal microbiota of the mammalian skin. It may appear in irregular clusters, groups of four or eight. Micrococcus luteus is a coccus shaped gram-positive bacteria. Micrococci may be distinguished from staphylococci by a modified oxidase test as shown below-. Members of the Staphylcocci can be differentiated based on production of the enzyme coagulase. 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]. Staphylococcus spp. Continuing without changing cookie settings assumes you consent to our use of cookies on this device. Biochemical Test and Identification of Staphylococcus aureus. We were aiming to isolate a bacterium from this initial environmental sample by using repeated quadrant streaks to isolate single species colonies, therefore producing a pure culture that we could do a multitude of tests on which include Gram staining, genetic analysis and antibiotic testing along with many other tests. M. luteus causes odours in humans when breaking down the components of sweat. Micrococci have Gram-positive spherical cells ranging from about 0.5 to 3 micrometers in diameter and typically appear in tetrads. Oral microbial flora of reptiles includes bacteria from Proteus, Porphyromonas, Micrococcus, Salmonella and Staphylococcus genera 5,8,15. In the case of a negative test, no colour change is observed. 2019). This kit utilizes enzymatic reactions to release bacterial DNA from the cell. It is a gram positive, coccus shaped microbe, and contains catalase. 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 also did a fluid thioglycolate test to determine the bacterias oxygen class. 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. Due to the thick structure of the bacterial cell wall, Gram-positive cocci retain the primary Grams stain, Crystal Violet. The Culture Collections represent deposits of cultures from world-wide sources. Table 1: Graph from Korona showing the percent reads of each organism, and to which taxonomic level. 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. The bacterium also colonizes the human mouth, mucosae, oropharynx and upper respiratory tract. To sample, I used sterile cotton swabs and streaked them on TSA plates. 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 organisms range in virulence potential from commensal organisms found on the skin and mucous membranes to pathogens associated with serious infections. 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. [8], Norwegian researchers in 2013 found a M. luteus strain that synthesizes a pigment that absorbs wavelengths of light from 350 to 475 nanometers. Micrococcus species are more sensitive to the Bacitracin while the staphylococci are resistant. This microbe is coccus shaped and forms in tetrads. Micrococcus luteus growth on nutrient agar is bright mustard-yellow colonies as shown above image. 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. Micrococcus luteus has been shown to survive in oligotrophic environments for extended periods of time. The genera Staphylococcus, Micrococcus and Sarcina were recognized. 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.

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