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Query: EC:3.5.1.5 (urease)
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A restricted biochemical scheme for the identification of enterobacteria, consisting of 12 enzymatic tests, of which 7 performed on the multitest TSI and MIU media (H2S, the production of acid and gas from glucose, fermentation of lactose/saccharose, mobility, urease and indol production) and 5 additional tests performed separately : lysindecarboxylase, phenylalanindeaminase, beta galactosidase, increase on citrate media and splitting of sodium malonate is proposed. Of 7782 coprocultures, 275 were selected on TSI and MIU media as belonging to one of the groups of known pathogenic enterobacteria ; 94.87% of these cultures were correctly identified by using the 5 additional tests alone. Of the 14 cultures that could not be listed taxonomically, 10 gave atypical reactions with at least one of these tests. The current use of this restricted scheme and the use of the more extensive sets only in doubtful cases presents a real advantage by reducing the volume of work and materials under satisfactorily accurate conditions for identification.
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PMID:[Value of some enzyme tests used in practice for identification of enterobacteria]. 14 13

Twenty-three isolates of Achromobacter species (CDC group Vd) were examined morphologically and biochemically. Gram stains revealed gram-variable bacilli frequently curved or hooked at one pole and often coryneform in shape and arrangement. Electron microscopy revealed the presence of extracellular material in polar accumulations and demonstrated the polar flagella arrangement seen by light microscopy to be lateral. Two colony types were produced; one was minute and watery at 24 h (35 degrees C) progressing to large, mucoid colonies at 48 h, and the other type was shiny, glistening, opaque but nonmucoid. All isolates grew on MacConkey agar and produced catalase, oxidase, and urease. Most grew on salmonella-shigella agar, reduced nitrate to nitrite and gas, hydrolyzed esculin, deaminated phenylalanine (2 to 4 days) and produced H2S in triple sugar iron agar (4 to 12 days). Oxidation of carbohydrates was weak, delayed, and limited to glucose and xylose. Two isolates also oxidized maltose, mannitol, and sucrose. The ability of miniaturized "nonfermenter" kits to identify Achromobacter species was tested. The Minitek (Baltimore Biological Laboratory, Cockeysville, Md.) and N/F (Corning, Roslyn, N.Y.) systems, respectively, identified 21 and 19 of the 23 isolates, whereas the Oxi/Ferm (Roche, Nutley, N.J.) identified 13 and the API 20E (Analytab Products, Plainview, N.Y.) identified only 3.
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PMID:Achromobacter species (CDC group Vd): morphological and biochemical characterization. 37 35

The Micro-ID system for rapid (4 h) identification of Enterobacteriaceae was evaluated by testing 433 enteric bacilli and 9 other gram-negative bacilli. Each isolate was identified with conventional tubed media and was also tested in the Micro-ID and API 20E systems. The overall accuracy of both systems was 97%. Micro-ID tests for the Voges-Proskauer reaction, indole and H2S production, and ornithine and lysine decarboxylase all demonstrated a 97 to 99% correlation with conventional methods. Only 86% of the Micro-ID urease tests agreed with Christenson urea agar. Two inoculum densities were tested in Micro-ID panels, with 157 stock cultures. Over 90% of the tests were unaffected by changes in inoculum density. Tests with four control strains suggested that the Micro-ID system was more reproducible when a light inoculum was used. The Micro-ID system was found to be a very convenient method for rapid, accurate, and precise identification of the Enterobacteriaceae.
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PMID:Rapid identification of Enterobacteriaceae with the micro-ID system versus API 20E and conventional media. 38 17

We have attended to a comparative research between two commercial microsystems: Enterotube and Minitek in order identify the Enterobacteriaceae and a reference system given by the combination of the usual macromethods already used in our laboratory. We have examined 401 bacterial cultures of Gram bacillus which we trought to belong to Enterobacteriums, coming from clinical material (excrements, urine, pharyngeal swabs, vaginal swabs, urethral swabs and espectoration) we have received for the bacteriological diagnosis. 390 of 401 cultures have shown to be Enterobacteriums. The biochemical reactions they have given show that the Enterotube and the Minitek have, with the usual system a good accordance for the following tests: dextrose (acid and gaz) lysin and ornithine decarboxylase, production of H2S and indole, phenylalanine deaminase and urease; while we have some statistically significant discordances for the fermenting of lactose and the use of citrate. We have also significant discordances E/C for the fermenting of dulcitole while the ones of Minitek are acceptables. The notes recommend the use, in the specialized bacteriological laboratories, of the conventional tests.
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PMID:[Evaluation of two miniaturized systems widely used in the laboratories for the identification of the "Enterobacteriaceae" (author's transl)]. 39 48

The possibility to keep some biochemical reactions of the parent strains (urease-positive, glucose fermentation, phenylalanine-deaminase-positive, H2S but not indol production) was demonstrated in 5 L-forms, obtained from as many strains of Pr. mirabilis and in 1 L-form, isolated from a vaginal secretion and identified as belonging to the same species. The indirect hemagglutination technique, made by the sonicated antigen in 3 of the 6 L-forms with Proteus OXK antiserum, resulted positive in titers varying from 1:128 to 1:1024. Crossed tests made with antisera for different bacterial species (e. coli, Shigella, klebsiella, ecc.) and of Mycoplasma (M. hominis, M. orale, M. salivarium, M. fermentans, M. arthritidis) put in evidence aspecific reactions only in 1.3% of the bacterial antisera. On the contrary, all 5 antisera for Mycoplasma were able to agglutinate the sensitized erythrocytes at titers quite analogous to that of the homologous antiserum. The sensitivities to various antibiotics of the 6 L-forms and the parent strains has been determined. All of L-forms were more resistent to the tetracycline than L-forms of other bacterial species. On the basis of te results got by biochemical and serological tests, we confirm the necessity to make use of both the groups of tests, in order to identify the L-forms of recent isolation.
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PMID:[Researches on some biochemical and serological properties and on the sensitivity to antibiotics of L-forms of "Proteus" (author's transl)]. 40 87

Twenty-two strains corresponding by their biochemical properties to the genus Levinea - Citrobacter were isolated. Six of the strains were referred to the species Citrobacter diversus and 12 to C. freundii, whose properties are identical with those of L. malonatica and L. amalonatica, respectively. Four strains differed from Levinea organisms by some reactions, but were fully compatible with C. freundii (in the scheme of EWING and DAVIS); two of them utilized malonate. The taxonomic position of strains displaying the following biochemical properties: dextrose positive, indole positive, H2S negative, urease on Christensen's citrate medium positive, lysine-decarboxylase negative-is discussed. In routine practice, these strains may be more accurately identified by adding of four tests: adonitol with gas production, KCN, raffinose and malonate.
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PMID:First isolations of Levinea-Citrobacter cultures in Czechoslovakia. 117 Jul

Twenty-one type or other reference strains, each representing a different Campylobacter, Helicobacter, or Arcobacter taxon, and a reference strain of Staphylococcus aureus were used to assess the reproducibility of nine enzyme detection tests used in the identification of campylobacters. For five of the tests (alkaline phosphatase, DNase, and H2S production, indoxyl acetate hydrolysis, and nitrate reduction), more than one procedure was employed to determine the most suitable method. Alkaline phosphatase test results were better defined and more reproducible if read after 1 h of incubation. Detection of DNase was fully reproducible with each method (except with Helicobacter pylori), but reactions were generally weaker than those of other DNase-producing organisms. Both procedures for determining H2S production were irreproducible for the same strains. The reproducibility of indoxyl acetate hydrolysis was improved by using disks impregnated with 25 microliters of substrate. Reduction of nitrate was best determined by Cook's plate method. Results for the other tests examined (catalase, oxidase, and urease production and hippurate hydrolysis) were both pertinent and fully reproducible for all strains.
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PMID:Assessment of enzyme detection tests useful in identification of campylobacteria. 155 96

On the basis of phenotypic characterization and DNA relatedness determinations, the genus Afipia gen. nov., which contains six species, is described. The type species is Afipia felis sp. nov. (the cat scratch disease bacillus). Afipia clevelandensis sp. nov., Afipia broomeae sp. nov., and three unnamed not associated with cat-borne disease. All but one strain (Afipia genospecies 3) were isolated from human wound and respiratory sources. All Afipia species are gram-negative, oxidase-positive, nonfermentative rods in the alpha-2 subgroup of the class Proteobacteria. They are motile by means of a single flagellum. They grow on buffered charcoal-yeast extract agar and nutrient broth, but rarely on MacConkey agar, at 25 and 30 degrees C. They are urease positive; but they are negative in reactions for hemolysis, indole production, H2S production (triple sugar iron agar), gelatin hydrolysis, esculin hydrolysis, and peptonization of litmus milk. They do not produce acid oxidatively from D-glucose, lactose, maltose, or sucrose. The major cell wall fatty acids are 11-methyloctadec-12-enoic (CBr19:1), cis-octadec-11-enoic (C18:1omega7c), and generally, 9,10-methylenehexadecanote and 11,12-methyleneoctadecanoate; and there are only trace amounts of hydroxy acids. The guanineplus-cytosine content is 61.5 to 69 mol%. A. felis is positive for nitrate reduction and is delayed positive for acid production from D-xylose, but it is catalase negative. A. clevelandensis is negative in all of these tests. A. broomeae is weakly positive for catalase production and acid production from D-xylose, but it is negative for nitrate reduction.
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PMID:Proposal of Afipia gen. nov., with Afipia felis sp. nov. (formerly the cat scratch disease bacillus), Afipia clevelandensis sp. nov. (formerly the Cleveland Clinic Foundation strain), Afipia broomeae sp. nov., and three unnamed genospecies. 177 49

A new microaerophilic, spirally curved, rod-shaped bacterium was isolated from the gastric mucosa of a pigtailed macaque (Macaca nemestrina). The gram-negative cells of this bacterium are oxidase, catalase, and urease positive and strongly resemble Helicobacter pylori (Campylobacter pylori) cells. Like H. pylori, this organism does not metabolize glucose, does not reduce nitrate or produce indole, does not produce H2S from triple sugar iron agar, does not hydrolyze hippurate or esculin, and does not grow in the presence of 1% glycine, 1.5% salt, or 1% bile. Also like H. pylori, it is resistant to nalidixic acid and susceptible to cephalothin. However, unlike H. pylori, the colorless colonies are flat and have irregular edges. This organism has a unique cellular fatty acid composition, forming a new gas-liquid chromatography group, group K, and a distinctive DNA content (24 mol% guanine plus cytosine). It exhibits less than 10% DNA-DNA homology (as determined by the nylon filter blot method at 65 degrees C) with other members of the genus Helicobacter. Although the levels of DNA relatedness between previously described Helicobacter species and the new organism are low (less than 10%) and the difference in guanine-plus-cytosine content is large (24 versus 36 to 41 mol%), the genus Helicobacter is the only genus in which it is logical to include the organism at this time. We propose that our single strain represents a new species, Helicobacter nemestrinae, and we designate strain T81213-NTB (= ATCC 49396) as the type strain.
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PMID:Helicobacter nemestrinae sp. nov., a spiral bacterium found in the stomach of a pigtailed macaque (Macaca nemestrina) 174 3

Over a 12-year period, 16 human strains of a gram-negative, catalase-positive, halophilic, aerobic, nonmotile, small coccoid bacterium were received for identification. On the bases of biochemical characteristics and cellular fatty acid profiles, 14 of these strains were similar to the "Philomiragia" bacterium (Yersinia philomiragia, species incertae sedis). Additional characteristics were growth on Thayer-Martin agar but no growth or sparse, delayed growth on MacConkey agar; oxidase positive; acid production, often weak and delayed, from D-glucose, sucrose, and maltose; urease negative; no reduction of nitrates; and H2S produced but often delayed in triple sugar iron agar. Both the human isolates and the "Philomiragia" bacterium contained C10:0, C14:0, C16:0, C18:1 omega 9c, C18:0, 3-OH C18:0, C22:0, and C24:1 as major cellular fatty acids and ubiquinone eight (Q8) as the major isoprenoid quinone. These cellular acids in these relative amounts have been found previously only in Francisella tularensis and Francisella novicida, suggesting a relationship between the "Philomiragia" bacterium and Francisella species. Of the 14 human "Philomiragia"-like isolates, 9 were from blood, 3 were from lung biopsies or pleural fluid, and one each was from peritoneal fluid and cerebrospinal fluid. DNA relatedness studies (hydroxyapatite method, 50 and 65 degrees C) showed that these 14 strains were a single group that was the same species as the "Philomiragia" bacterium. Two other human strains were oxidase negative and H2S negative. They formed a single DNA relatedness group that was indistinguishable from the type strains of both F. tularensis and F. novicida. DNA relatedness of "Philomiragia" bacterium type and other strains to strains of F. novicida and F. tularensis, including the type strains, was 35 to 46%. One of the two F. novicida- and F. tularensis-like strains was isolated from blood, and the other was isolated from a cervical lymph node. On the basis of these findings, we propose transferring Y. philomiragia from the genus Yersinia to the genus Francisella as Francisella philomiragia comb. nov. Having confirmed that F novicida and F. tularensis are the same species and having shown that F. novicida is pathogenic for humans, we further propose eliminating the species F. novicida and demoting it to a biogroup of F. tularensis.
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PMID:Francisella philomiragia comb. nov. (formerly Yersinia philomiragia) and Francisella tularensis biogroup novicida (formerly Francisella novicida) associated with human disease. 267 Oct 19


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