Detection of bacterial phosphatase activity by means of an original and simple test.

Satta, G.; Grazi, G; Varaldo, Pietro E.; Fontana, Roberta

doi:10.1136/jcp.32.4.391

Cited by 33 publications

(21 citation statements)

References 16 publications

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“…A heat-killed cell suspension of M. luteus was prepared and added to the melted media as described previously (14). The same procedure was employed with the other strains tested as substrates, the only modifications being in the temperature and time of incubation, as needed, to obtain optimal growth.…”

Section: Methodsmentioning

confidence: 99%

Grouping of Staphylococci on the Basis of Their Bacteriolytic-Activity Patterns: A New Approach to the Taxonomy of the Micrococcaceae: I. Identification of Six Different "Lyogroups"

Varaldo

Satta

Grazi

et al. 1978

International Journal of Systematic Bacteriology

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As a further development of our previous finding that heterogeneous bacteriolytic activities are produced by virtually all staphylococcal strains, several different conditions were tested for their selective and differentiating interferences with the bacteriolytic activity of some staphylococcal strains. Such a study has indicated the possibility of resolving different patterns of lytic activity among staphylococci and has resulted in the preparation of an assay system, composed of eight testing media, for lytic enzymes. A total of 1,054 staphylococcal strains has been tested on these media and found to fall into six "lyogr~ups,'~ each characterized by a specific pattern of bacteriolytic activity. The novelty and taxonomic relevance of analyzing the bacteriolytic activity of staphylococci and the possibility of extending the proposed method to bacteria other than staphylococci are discussed.There is not yet general agreement upon both the definition of the genus Staphylococcus, in particular its separation from the genus Micrococcus, and its subdivision into species or biotypes. Only minor taxonomic problems exist in dealing with coagulase-positive staphylococci. Staphylococcus aureus is, in fact, unanimously considered a well-defined and homogeneous species, at least as far as strains of human origin are concerned (3,5,8). Most of the problems are concerned with coagulase-negative staphylococci, for these strains are rather heterogeneous, and the schemes proposed for their classification and identification only partially coincide (1,4,5, 9, 11, 16, 17).The discovery of some new physiological property that varies with the different groups of staphylococci would be helpful in resolving these problems.We have recently shown (14) that under suitable conditions virtually all staphylococci produce an easily detectable bacteriolytic activity. We also found that the optimal conditions for expressing bacteriolytic activity differ not only between coagulase-positive and coagulase-negative staphylococci, but also between different coagulase-negative strains. Therefore, bacteriolytic activity seemed to us to be a good candidate for a characteristic of high taxonomic value.To test for such a possibility, we studied the bacteriolytic activity of 1,054 staphylococci under various conditions of growth. As a result, we were able to subdivide these strains into six different groups, which we have named "lyogroups" ( X k v = to lyse), the members of each group producing a particular type of lytic activity. It will be shown in an accompanying paper (18) that strains sharing a particular bacteriolytic activity also share a great majority of other biochemical and physiological characters. MATERIALS AND METHODSBacterial strains. A total of 1,054 staphylococci from humans were examined in this study. Most were isolated from clinical materials-blood, urine, sputum, pus, throat swab, etc.-examined in the Clinical Bacteriology Laboratory of the Institute of Microbiology of the University of Genoa Medical School. Some strains were isolate...

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Section: Methodsmentioning

confidence: 99%

Grouping of Staphylococci on the Basis of Their Bacteriolytic-Activity Patterns: A New Approach to the Taxonomy of the Micrococcaceae: I. Identification of Six Different "Lyogroups"

Varaldo

Satta

Grazi

et al. 1978

International Journal of Systematic Bacteriology

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“…In some experiments, bacterial growth phosphatase activity was also determined by the methyl green-phenolphthalein diphosphate (MGP) method previously described (31,33,34). Briefly, to the melted media methyl green (Carlo Erba, Milan, Italy) and phenolphthalein diphosphate (Sigma), sterilized by filtration, were added at final concentrations of 25 and 1,000 mg/ml, respectively, from 100-fold-concentrated stock solutions.…”

Section: Methodsmentioning

confidence: 99%

“…However, it has not yet been extensively analyzed for micrococci. From the few studies performed, it was concluded that micrococci do not have this property (3,11,16,21,31). However, in studying the phosphatase activity of the major staphylococcal species, some of which had previously been reported to be positive for this feature but most of which had been reported to be negative (3,16,37), we have recently shown that all staphylococcal species analyzed demonstrated phosphatase activity (40).…”

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confidence: 99%

Micrococci Demonstrate a Phosphatase Activity Which Is Repressed by Phosphates and Which Can Be Differentiated from That of Staphylococci

Satta

D’Andrea

Grazi

et al. 1993

International Journal of Systematic Bacteriology

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The phosphatase activities of 114 micrococcal strains belonging to seven different species and of an additional 150 unspeciated micrococcal strains were evaluated on solid media at various pHs containing or not containing phosphates. In the presence of phosphates, only nine strains (five unspeciated strains, one Micrococcus luteus strain, and three Micrococcus varians strains) yielded a positive reaction on plates at pH 8. In media (at pH 8) deprived of phosphates, in contrast, all but 15 strains demonstrated clear-cut phosphatase activity. Acid phosphatase could not be evaluated on solid media since none of the strains grew satisfactorily on plates at pH 5. The phosphatase activities of seven (one or two for each species, which included phosphatase-negative strains) of the strains whose colonies proved phosphatase negative at pH 8 and of 18 (two or three strains per species) of those with phosphatase-positive colonies were evaluated at pH 5 and 8.5 in toluene-treated cells which had been grown in liquid media at pH 7 containing or not containing phosphates. All strains demonstrated distinct phosphatase activity at both pHs when grown in media not containing phosphates. In contrast, when strains were grown in the presence of such substances, virtually no activity was observed at pH 8.5, and, generally, a much reduced activity was observed at pH 5. The phosphatase activity of micrococci of the various species (three to eight strains per species) was also compared with that of staphylococci of different species (5 to 10 strains per species) by the methyl green-phenolphthalein diphosphate method, the sensitivity of which can be varied by using different enzyme substrates. By using phenolphthalein diphosphate as an enzyme substrate, it was found that virtually all the different species of staphylococci yielded a positive reaction on plates not containing phosphates while almost all micrococci proved phosphatase negative with the methyl green-phenolphthalein diphosphate method. This indicates that the phosphatase activity of micrococci can easily be differentiated from that of staphylococci.Of the metabolic properties generally exploited for defining and characterizing the different bacterial species, such as enzyme activities and sugar fermentation, only a few have so far been identified in micrococci. This leaves us with relatively few tests that, particularly in some species (see reference 17), can be used for characterizing the various strains of the group. This is rather unfortunate since it makes micrococcal species definition and identification difficult, particularly with the standard type of assays generally performed in routine clinical microbiology. However, it is likely that further studies may lead to identification of additional metabolic properties that may prove useful for characterization of this bacterial group.Phosphatase activity is a property often evaluated as a test for species definition and identification (3,10,12,15,48). However, it has not yet been extensively analyzed for micrococci. From ...

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“…Coagulase activity was assayed as described elsewhere (17) by adding 0.2 ml of pure SaG preparation to 2 ml of rabbit plasma. Protease, lipase, phosphatase, and heat-stable DNase were assayed as described (18)(19)(20)(21), respectively, by depositing 0.1 ml ofpure SaG preparations (sterilized by filtration) in adequate wells prepared in the specific solid media. Positive controls were represented by protease type I from bovine pancreas, lipase type VI-S from porcine pancreas, phosphatase type I from bovine intestine, and deoxyribonuclease I type II from bovine pancreas (Sigma Chemical Co., St. Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

Staphylococcal endo-beta-N-acetylglucosaminidase inhibits response of human lymphocytes to mitogens and interferes with production of antibodies in mice.

Valisena¹,

Varaldo²,

Satta³

1991

J. Clin. Invest.

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Detection of bacterial phosphatase activity by means of an original and simple test.

Cited by 33 publications

References 16 publications

Grouping of Staphylococci on the Basis of Their Bacteriolytic-Activity Patterns: A New Approach to the Taxonomy of the Micrococcaceae: I. Identification of Six Different "Lyogroups"

Grouping of Staphylococci on the Basis of Their Bacteriolytic-Activity Patterns: A New Approach to the Taxonomy of the Micrococcaceae: I. Identification of Six Different "Lyogroups"

Micrococci Demonstrate a Phosphatase Activity Which Is Repressed by Phosphates and Which Can Be Differentiated from That of Staphylococci

Staphylococcal endo-beta-N-acetylglucosaminidase inhibits response of human lymphocytes to mitogens and interferes with production of antibodies in mice.

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