micrococcus luteus urease test

Nuclease production and lysostaphin susceptibility of, Evaluation of the Staph-Zym system for identification of, Biochemical and serological properties of. Resistance of E. gallinarum,E. abstr. Evaluation of a micromethod gallery (API Staph) for the identification of staphylococci and micrococci. API 20E, MicroScan conventional overnight, and Vitek GNI Card include growth-dependent tests and a few enzyme tests; in general, they do not use the ability of enzyme tests to provide results rapidly. Composition and differentiation of the genus, Glycosidase profiles of members of the family. They are positive for catalase and oxidase (modified). Raus and Love (189) compared the activities of staphylocoagulases from S. aureus and S. intermedius by using Chromozym-TH and concluded that S. intermedius staphylocoagulase resembled S. aureusstaphylocoagulase in its rate and mode of action on prothrombin but that the enzyme was produced in lesser amounts in the former species. Staphylococcus is the only genus showing susceptibility to lysostaphin, but some exceptions occur among species ofMicrococcus and Staphylococcus. Evaluation of the Rapid Strep System for identification of gram-positive, catalase-negative cocci isolated from bovine intramammary infections. Predictive values of species identifications from the Vitek Gram-Positive Identification card using clinical isolates of coagulase-negative staphylococci. Substrates for separation within the genus Micrococcusincluded hydroxyprolyl, glycyl-prolyl, aspartyl, and tyrosyl-seryl conjugates of naphthylamine. Tests based on rapid latex agglutination and hemagglutination to detect clumping factor or protein A are also used extensively (79, 131); however, Fournier et al. sedentarius, and M. lylae were included in the studies by Baldellon and Mégraud (12). In general, many systems performed better when augmented with serological and other tests (14, 252). Members of the genus (such as L. citreum, L. lactis, L. mesenteroides, L. pseudomesenteroides, and L. paramesenteroides) are generally found on vegetable material, in milk products, and in other fermentation products such as wine and sausages. Multicenter comparison of MicroScan Rapid gram-positive combo panel 1 and positive combo panel 5 with conventional methods for identification of. They grow in circular, entire, convex and creamy yellow pigmented colonies with diameters of approximately 4 mm after 2-3 days at 37°C. The presence of catalase is usually determined by addition of a drop of 3% H2O2 to a heavy bacterial suspension and observation of effervescence due to the release of O2. They do not provide expected results and are less rigorously regulated. The use of API enzyme research kits detecting 20 glycosidases, 10 esterases, 57 arylamidases, alkaline and acid phosphatases, and phosphoamidase has been reported (151, 153, 164, 226). Isolates of V. fluvialis resemble members of the genus Enterococcus both phenotypically (68) and genetically (42), reacting positively with the AccuProbe Enterococcus test, but unlikeEnterococcus spp., they grow poorly at 45°C. faecium, was separated from Streptococcus on the basis of DNA hybridization data by Schleifer and Kilpper-Bälz (208). The results were observed after 3 h of incubation and were combined with a 48-h test for the determination of hyaluronidase production. (93) suggested the usefulness of a rapid PYR test with a β-naphthylamine conjugate of pyrrolidonyl for the identification of group A streptococci and enterococci. The MicroScan Rapid Pos Identification system is the only one relying on enzyme tests and acid formation detected fluorometrically and providing identification of most clinically significant staphylococci, enterococci, and streptococci in 2 h. Micrococcus.The members of the genusMicrococcus differ from those of Staphylococcusby being obligate aerobes, with a G+C content of 63 to 73 mol%, containing cytochromes a, b, c, andd, lacking teichoic acids in their cell walls, lacking glycine in the interpeptide bridge of their cell walls, being resistant to furazolidone, and being susceptible to bacitracin. Susceptibility tests . The time is needed for selection of a sufficient number of isolates for testing, finding tests to differentiate the newly described taxa from existing ones, establishing the manufacturability of required tests, performing appropriate testing of the modified product with further isolates, and obtaining regulatory approval of the modified kit. Comparison of the API Staph-Ident and DMS Staph-Trac systems with conventional methods used for the identification of coagulase-negative staphylococci. These could be attributed to the differing pH conditions of the tests and possibly also to the difference in the affinity of the enzymes to the synthetic moiety of the substrate. A specific enzyme test for β-galactosidase based on utilization of the synthetic substrateo-nitrophenol-β-d-galactopyranoside (ONPG) or substrates with other synthetic moieties is included in most kits for identification of the Enterobacteriaceae. Rapid biochemical tests for the identification of groups A, B, C, F, and G streptococci from throat cultures. Comparison of accuracy of identification systems forStreptococcaceae and similar organisms. C-296. DNA-DNA hybridization studies and phenotypic characteristics of strains within the “, Comparison of five agglutination tests for identification of. Devriese et al. (36) confirmed that the enzyme is liberated into the medium in the early logarithmic phase (3 to 6 h), and that its production depends on aeration and a rich growth medium such as brain heart infusion. The substrates were used in liquid medium and incorporated in a selective agar medium. Relationships between results obtained by conventional and enzyme tests have been scantily studied, but quite often these sets of results provide related information. It is a gram positive, coccus shaped microbe, and contains catalase. (64); they have found that neither the conventional test system nor the API Rapid Strep identification system could differentiate between the two species and recommended comparison of whole-cell protein patterns for that purpose. Preliminary evaluation of Biolog, a carbon source utilization method for bacterial identification. They also found that the presence of certain saccharides could inhibit the hydrolysis of certain fluorogenic substrates and that 1% glucose inhibited the hydrolysis of conjugates of α-d-galactose and α-l-arabinose by E. faecium. They can cause infections at a wide variety of sites, including the urinary tract, bloodstream, endocardium, abdomen, and biliary tract, as well as burn wounds and indwelling devices (114). Simplified scheme for routine identification of human. M. kristinae, M. luteus, M. varians, M. nishinomiyaensis,M. aureus) is used in the commercially available RAPiDEC Staph kit. Littel and Hartman (148) tested 44 fluorogenic substrates for their ability to differentiate between fecal enterococci and streptococci. STAPHYtest (Lachema, Brno, Czech Republic), a microtiter-based system with nine tests and with eight compartments per isolate for identification of Micrococcus,Stomatococcus, and Staphylococcus, has been described (211). The genus represents a distinct line of descent quite separate from aerococci and pediococci (46). The commercial identification systems also provide databases of expected results, and an unknown isolate is assigned to one of the taxa in the database either by using a code book or by using an automated system and computer-based identification. (136) and Baldellon and Mégraud (12) examined strains from members of the Micrococcaceae by using a variety of API test strips. In our lab, we only use the urease broth, not the agar. (i) Gemella.The genus Gemellacontains two species G. haemolysans, previously classified as Neisseria haemolysans, and G. morbillorum, previously classified as Micrococcus sp., Diplococcus morbillorum, Peptostreptococcus morbillorum, andStreptococcus morbillorum (25, 196). The current division, based on 16S rRNA sequencing (24), shows that the pyogenic streptococci cluster in one phylogenetic group (group I), which includes the above taxa except for S. anginosus and S. sanguis, which are now included in the viridans group (group III). (29) have described 19 isolates of this species from ear fluid samples. Important: Moreover, some of the newly described taxa have originated from phylogenetic studies of as little as one isolate and may exhibit only genetic and no phenotypic differences from existing taxa. Taxonomic studies on some leuconostoc-like organisms from fermented sausages: description of a new genus. The clinical significance of the two species has been reviewed (68). Evaluation of Staph ID 32 system and Staph-Zym system for identification of coagulase-negative staphylococci. Collins et al. The broth contains two pH buffers, urea, a very small amount of nutrients for the bacteria, and the pH indicator phenol red. Effect of BiTek Agar on lysostaphin susceptibility of staphylococci. In addition to the schemes listed in the tables, Watts et al. Micrococcus Morphology: - Gram +ve cocci - Arrangement : Tetrades - Non motile, non capsulated, non sporulated Habitat: May be normal present in upper respiratory tract Species : 1-M.varians 2- M. luteus 3- M.roseus Culture: - Strictly aerobic at 37°C incubation (24 hr) - Grow on ordinary media Nutrient agar - … An indicator in the buffer changes color if the bacteria are able to grow (i.e., no antibiotic is present). Enzyme tests in bacterial identification. Our study fills in the gap in the molecular database on Kocuria rhizophila and Micrococcus luteus strains obtained from moribund fish and supplements the knowledge concerning its pathogenic properties for salmonids. (62) have shown that testing for β-d-glucosidase activity in the presence of 2.5% sodium deoxycholate gives results equivalent to the conventional bile-esculin test. The largest number is available for the identification of clinically important aerobic and facultatively anaerobic gram-negative bacteria (e.g., API 20E, MicroScan conventional overnight and MicroScan Rapid GN Identification Systems, and Vitek GNI Card). Arylamidase activities were found in most species and could be used to separateMicrococcus from the other genera as well as to differentiate among some Micrococcus species. In the presence of the reagents and acetoin, a cherry-red color develops. Phylogenetically, the genus is related to Aerococcus; the G+C content of the type strain is 37 mol% (41). When susceptibility is tested by observation of growth inhibition zones, it also depends on the medium on which the bacteria are grown (144). Isolates grow very slowly in 6.5% NaCl, are LAP and PYR positive, are susceptible to vancomycin, do not produce gas from glucose, and do not form acid from carbohydrates. The second species,V. However, a number of factors determine whether the two tests will provide identical information. The former enteric and lactic streptococci are now placed in the separate genera Enterococcus and Lactococcus, respectively. The remaining species have been divided, based on 16S rRNA sequencing, into six groups by Bentley et al. Isolates assigned to this genus are obligately aerobic, slow-growing cocci that give a weak reaction in the catalase test but do not contain cytochrome. Isolates are positive for the LAP and bile-esculin tests but negative for PYR. The benzidine test (56) detects iron-porphyrin compounds in catalase- and cytochrome-containing bacteria; it is highly sensitive and is positive for all members of the Micrococcaceae, even those which are catalase negative. Description and evaluation of the semiautomated 4-hour rapid ID 32 Strep method for identification of streptococci and members of related genera. Vandenesch et al. In further studies,S. They showed that the staphylocoagulase-prothrombin complex can hydrolyze the chromogenic substrates tosyl-Gly-Pro-Arg-p-nitroanilide (Chromozym-TH), Z-Gly-Pro-Arg-p-nitroanilide, H-d-Phe-Pip-Arg-p-nitroani- lide, and tosyl-arginine-methyl ester (TAME); it can also hydrolyze the fluorogenic substrate Boc-Val-Pro-Arg-4-methylcoumaryl-7-amide. In the 1970s and 1980s, enterococci became firmly established as major nosocomial pathogens. Only S. gallinarum, S. lentus, and S. sciuri can hydrolyze esculin and are positive for the β-glucosidase test. They reported that β-naphthylamine substrates inhibited the growth of streptococci. Resistance to vancomycin and teicoplanin: an emerging clinical problem. Enter multiple addresses on separate lines or separate them with commas. Contents Schemes for differentiating the members of this genus from the other gram-positive cocci have appeared (68). A modification of the benzidine test has been described (75) for the detection of oxidase-positive members of theMicrococcaceae based on removal of noncovalently linked heme groups prior to reaction with the benzidine reagent making it specific to the covalently protein bound heme of cytochromes c. The modified benzidine test was compared with a modified oxidase test for differentiation within the Micrococcaceae and shown to be positive for all members of the genus Micrococcus and for strains of S. sciuri. Previously, the genus Micrococcus contained nine species, and tests that may help in differentiation between the species have been described (129). Many enterics can hydrolyze urea; however, only a few can degrade urea rapidly. The early recognition of the usefulness of the PYRase test for differentiation of streptococci may be responsible for the large number of enzyme tests included in these kits. Dealler et al. Flynn and Ruoff (77) described results obtained with a rapid commercially available system, Fluo-Card Milleri (Key Scientific, Round Rock, Tex.) The clinical and economic importance of members of these taxa is briefly summarized. Members of the genus have a G+C content of 34 to 46 mol% and are pathogenic for humans and other animals; some species are found as members of the normal flora of the mouth and gastrointestinal tract. Recently, a fourth type of resistance mediated by VanD has been described (146). Isolates of both taxa grow poorly with variable hemolysis, which may depend on the source of blood (rabbit, sheep, or horse). Coagulase-negative staphylococci and the epidemiological typing of, Simplified method for the isolation, identification and characterization of, Rapid identification of Gram-positive cocci on MicroScan fluorogenic plates from blood culture broth, abstr. Evaluation of MicroScan Rapid ID/MIC vs. Vitek Senior in a University Medical Center Clinical Setting, abstr. E. sulfureus has been isolated from plants (111). They exhibit a high G+C content of 56 to 60 mol%, near to that of members of Micrococcus (64 to 75 mol%) and unlike that of staphylococci (30 to 39 mol%). Reproducibility of API Staph-Ident system identifications of coagulase-negative staphylococci isolated from blood. Evaluation of RapiDEC Staph for identification of, Comparative evaluation of the Vitek Gram-positive identification system and the API Rapid Strep system for identification of. The difference may be attributed to the longer incubation period used in the agar test. A classification of micrococci and staphylococci based on physiological and biochemical tests. Classification and identification of the Viridans streptococci. milleri” group, which is based on the determination of three of the above mentioned-glycosidases, namely, α- and β-glucosidase and β-fucosidase, with MEU derivatives. Results: 11-16-15 Gram + is positive and Gram – is negative for the presence of acetoin in glucose fermentation. False-negative results in the tube and slide coagulase tests have been reported for 8 and 17 strains, respectively, of 87 strains of S. aureus. Novel method for rapid identification of Nocardia species by detection of preformed enzymes. Unfortunately, serological techniques relied upon previously for the identification of pyogenic streptococci identify isolates with group B and F antigens but not individual species of isolates positive for group A, C, and G antigens (197, 224). Among the enzymes used for the differentiation of these taxa, PYRase is the most common. Difficulties in the identification of isolates from veterinary sources have been observed with most systems. Evaluation of a rapid method for the determination of, Pyrrolidonyl peptidase in bacteria: a new colorimetric test for differentiation of. The API ZYM system (API System; bioMérieux, Paris, France) is a semiquantitative micromethod designed for the detection of 19 enzymatic activities (106, 232). 16S rRNA sequence determination for members of the genus. Schemes for the differentiation of members of the family have been published (12, 129, 194). Planococcus.Members of the genusPlanococcus are cocci arranged in pairs or tetrads. The relationship between classification results obtained by different molecular genetic techniques should also be considered. Strains have been isolated from patients with bacteremia, urinary tract infections, and meningitis. abstr. Although different numbers of isolates were tested (177 and 965, respectively) the results for α-galactosidase were 4 and 31% for the Hartley digest and Columbia-based agars respectively; the results for acid formation from raffinose were 20 and 40%, respectively. (24) suggested that the organisms belong in three different phylogenetic groups. Peel et al. Most systems are fairly successful in differentiating S. aureus, S. epidermidis, andS. Evaluation of two commercial systems for identification of coagulase-negative staphylococci to species level. The MAST ID system (Mast Laboratories Ltd., Bootle, England) provides a means of determining the metabolic activities of a number of isolates by agar plate and multipoint inoculator techniques (84, 86, 123, 193). (225) demonstrated that both subspecies of S. schleiferi can promote clotting of rabbit plasma in the standard tube test for coagulase. Is enterococcus faecalis positive or negative with the urea broth test? Inc.); a chromogenic test for the same arylamidase, involving a β-naphthylamine conjugate of pyroglutamic acid, is incorporated in the Pasco Gram-Positive ID Panel. The ability to identify S. bovis rapidly is important, since this species is associated with endocarditis and colon cancer, as reviewed by Coykendall (49). milleri” group into three species, S. anginosus, S. constellatus, andS. The relationships between the results of conventional biochemical and rapid enzyme tests are described and considered. milleri” isolates; the latter isolates give negative results. Comparison of Vitek Gram-positive identification system with API Staph-Trac system for species identification of staphylococci of bovine origin. Evaluation of the Automicrobic System for the identification of. In addition, the isolation of vancomycin-resistant bacteria from human infections (37, 48, 115, 200) has necessitated the development of tests and schemes for the identification of clinical isolates ofLeuconostoc and Pediococcus. jill_uebele. Streptococci and “Streptococcus-like” bacteria: old friends and new species. pyogenes based on the detection of PYR and β-glucosidase with β-naphthylamide and indoxyl conjugates, respectively. Separation on the basis of biochemical tests between beta-hemolytic isolates of antigenic groups C and G has been somewhat difficult. Evaluation of two fluorogenic assays for identification of. Take a clean, scratch free glass slide. Identification of coagulase-negative staphylococci with the API Staph System. Leuconostoc isolates are positive for the production of CO2 from glucose and negative for arginine dihydrolase, PYR, and LAP; 31% of the strains from clinical sources react with the group D antiserum (68). Oberhofer (173) found that all 26 strains of S. haemolyticus, the single strain of S. intermedius, and 2 of 7 S. warneri strains tested were positive in the pyrrolidonyl-arylamidase (PYR) test, while 65 isolates of S. epidermidis, 7 isolates of S. hominis, 8 isolates of S. saprophyticus, and 2 isolates of S. capitis were negative. With this almost universal range of survivable living conditions that microbes can live in (particularly bacteria), it would be reasonable to assume that there would be at least one variety living i… Comparison of Pasco and MicroScan Gram-positive ID Systems, abstr. The AccuProbe Enterococcus DNA culture confirmation probe (Gen-Probe, San Diego, Calif.) has been recommended for positive identification of all enterococci because most species of enterococci react positively with the probe with the exceptions of the type strains of E. cecorum, E. columbae, and E. saccharolyticus (73). Laboratory diagnosis Microdase (Modified Oxidase) Test 1. Porphyrin test as an alternative to the benzidine test for detecting cytochromes in catalase-negative gram-positive cocci. The acquisition of resistance to vancomycin, used for treating infections caused by gram-positive cocci that are resistant to other drugs, has been on the increase since the late 1980s (170). They showed that most species did not exhibit production of acid from the 49 carbohydrates included in API 50CH kit. There are a number of identification kits, some covering the facultative gram-positive cocci (e.g., MicroScan GP and Rapid GP panels and Vitek GPI Card) and others being specific for streptococci (e.g., API 20 Strep). 347, Evaluation of the MicroScan Rapid-pos ID panel. Differences between the results for the same nominal enzyme in different API kits have also been observed (127). Synthetic substrates for the detection of endopeptidases and lipases are available but are more difficult to incorporate into a dried, easily reconstitutable format. Microorganisms have been classified and identified on the basis of a variety of characteristics including morphological, growth, tolerance, metabolic, biochemical, and genetic. Comparison of physiologic tests used to identify non-beta-hemolytic aerococci, enterococci, and streptococci. Lack of lytic activity is confined to a small number of isolates of S. xylosus (205, 228). Identification of staphylococci with a self-educating system using fatty acid analysis and biochemical tests. (iii) Pediococcus.The genusPediococcus, with a G+C content of 34 to 43 mol%, contains five species associated with lactic acid fermentations of vegetables, grain mashes, and cheese (P. acidilactici, P. damnosus, P. dextrinicus, P. parvulus, andP. Completion times for the identification of bacteria taken from isolated colonies can vary from 2 h to several days. A computer-assisted algorithm is used to determine final identifications. Acid formation from glycerol, galactose, sorbitol, and d-tagatose differentiates between the two species; the former is positive in the first three tests, and the latter is positive only in the last (233). Reports on the identification of streptococci and related organisms with commercial kits are summarized in Table8. Applications of genetic methods (24, 41-46, 209, 244, 248), numerical taxonomy (31, 186), and enzymatic techniques (22, 127) have contributed to a further understanding of this complex taxonomic group. E. dispar, E. hirae, E. flavescens, E. mundtii, and E. raffinosushave been isolated from humans and other sources (73, 111).E. (iii) Lactococcus.The genusLactococcus includes the cocci of serogroup N that produce lactic acid as the main fermentation product; it can be confused withEnterococcus and differs from it by its antigenic reaction and its lesser ability to grow at 45°C. G. haemolysans may appear gram variable or gram negative. However, while the ONPG test requires only these two processes, acid formation from lactose requires the conversion of the glucose and galactose molecules into lactic and acetic acids, etc. Of the genera included in the family Micrococcaceae,Staphylococcus is the most clinically important and contains 32 species, most of which live on the skin and other external surfaces of animals (129). Rapid identification of coagulase negative Staphylococci species associated with veterinary infections. Enzyme fluorescence procedure for rapid diagnosis of streptococcal pharyngitis. Isolates are characterized by homo- or heterofermentative metabolism of carbohydrates. A number of tests have been used to differentiate between members of the Micrococcaceae andStreptococcaceae. The identification of the less common species is more variable. A simplified system for the identification of staphylococci by multipoint inoculation of test media. Most strains are positive in the LAP and PYR tests but require a heavy inoculum. However, the equivocal taxonomy of the viridans streptococci has hindered the development of identification kits with the required accuracy for this group. Specificity study of kits for detection of group A streptococci directly from throat swabs. This is due to reclassification of some species of Micrococcus. Characterization of ear fluid isolates of, Increasing prevalence of methicillin-resistant. Some may be applicable to isolates taken from different environments (143, 240). • *urea hydrolysis test (urease) • *phenylalanine deaminase test • *H2S test • *motility-indole-ornithine/MIO test ... Micrococcus luteus Micrococcus roseus Mycobacterium phlei Sporosarcina ureae Staphylococcus aureus Staphylococcus epidermidis Streptococcus pyogenes Streptococcus salivarius.

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