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PubMed1995

[Characterization of bacteriophage nontypable patterns of S. aureus strains].

Out of 195 strains of S. aureus isolated from in and outpatients. 43 were proven not to be lysed by the chosen set of bacteriophages. Of these nontypeable strains 14 were isolated from blood cultures and septic wounds of patients from Poznań University Hospital. 29 strains were derived from the anterior nares or throat swabs of outpatients, who were treated for chronic pharyngitis in the children's clinic in Poznań. S. aureus was identified using standard bacteriological method. Sensitivity tests to antibiotics were performed using the ATB computer system (bioMérieux). Beta lactamase production was examined using the iodometric method. The intracellular esterases of S. aureus were separated by polyacrylamid gel electrophoresis and stained with Fast Blue using alpha-naphtyhl acetate, beta-naphthyl propionate and indoxyl acetate as substrates. A much higher percentage of S. aureus strains isolated from specimens obtained from the hospital patients showed the ability to produce beta-lactamase than the strains cultured from outpatients. The esterase patterns of S. aureus are helpful in differentiating bacteriophage nontypeable strains.

Adolescent↗
PubMed1995

[Bacteriophages of serologic group B converting synthesis of fibrinolysin in S. aureus].

On the basis of the length of tail, two morphological types of S. aureus bacteriophages of the serogroup B converting fibrinolysine were shown. The properties of the 22 S. aureus bacteriophages of the serologic group B were investigated. All of the investigated bacteriophages were classified as the morphological group II of the family Styloviridae on the basis of electron microscope analysis. The size of capsids of the examined bacteriophages was 53.85 +/- 1.05 nm and the tail length varied from 135.4 nm to 170.5 nm. All of them had the tail terminated in the basal plate. The lytic properties of the investigated bacteriophages were not identical. 7 of them showed features of lytic group I and 15 of lytic group III. The members of lytic group III had apparently a longer tail than bacteriophages of lytic group I.

Fibrinolysin↗
PubMed1995

[Bacteriophages of serologic group F converting synthesis of fibrinolysin and beta toxin in S. aureus].

The properties of the 18 S. aureus bacteriophages of the serogroup F were investigated. The chosen bacteriophages were able to convert production of fibrinolysine inhibited synthesis of beta toxin. All of the investigated bacteriophages were classified as morphological group II of the family Styloviridae on the basis of the electron microscope analysis. The size of capsids of the examined bacteriophages was 54.10 +/- 0.80 nm and the tail length was from 206.1 nm to 311.9 nm. Most of them (15 bacteriophages) had the tail terminated in the basal plate. The lytic properties of the investigated bacteriophages were not identical. 11 of them showed features of lytic group III and 7 of lytic group V (miscellaneous).

Enterotoxins↗
PubMed1997

[Lysogenic conversion as a factor influencing tolerance to vancomycin in Staphylococcus aureus].

The standard, non-lysogenic, bacteriophage-free S. aureus NCTC 8325-4 strain was lysogenized with 15 different, obtained in our laboratory staphylokinase-converting bacteriophages belonging to serological groups A, B and F. MIC and MBC of vancomycin as well as the ratio of MBC to MIC were evaluated for all 15 lysogenic derivatives. The obtained results were compared with those for maternal strain. In the case of eight strains the ratio MBC/MIC showed the presence of tolerance to vancomycin (MBC/MIC > or = 32). Four of the vancomycin-tolerant derivatives were lysogenized with bacteriophages belonging to the serological group A, two were members of group B and two belonged to group F.

Anti-Bacterial Agents↗
PubMed1996

Construction of a chromosome map for the phage group II Staphylococcus aureus Ps55.

The genome size and a partial physical and genetic map have been defined for the phage group II Staphylococcus aureus Ps55. The genome size was estimated to be 2,771 kb by pulsed-field gel electrophoresis (PFGE) using the restriction enzymes SmaI, CspI, and SgrAI. The Ps55 chromosome map was constructed by transduction of auxotrophic and cryptic transposon insertions, with known genetic and physical locations in S. aureus NCTC 8325, into the Ps55 background. PFGE and DNA hybridization analysis were used to detect the location of the transposon in Ps55. Ps55 restriction fragments were then ordered on the basis of genetic conservation between the two strains. Cloned DNA probes containing the lactose operon (lac) and genes encoding staphylococcal protein A (spa), gamma hemolysin (hlg), and coagulase (coa) were also located on the map by PFGE and hybridization analysis. This methodology enabled a direct comparison of chromosomal organization between NCTC 8325 and Ps55 strains. The chromosome size, gene order, and some of the restriction sites are conserved between the two phage group strains.

Bacteriophage Typing↗
PubMed2002

Reaction difference rule for phage typing of Staphylococcus aureus at 100 times the routine test dilution.

Duplicate phage typing tests of 150 isolates of methicillin-sensitive and -resistant Staphylococcus aureus at 100 times the routine test dilution showed that the existing reaction difference rule remains satisfactory when isolates are tested on the same day; i.e., two or more reactions must differ before isolates should be considered distinct. However, if typing is done on different days, a three-reaction-difference rule is needed.

Bacteriophage Typing↗
PubMed1978

Comparison of "invasive" and "non-invasive" strains of Staphylococcus epidermidis by phage typing.

Seventy strains of coagulase-negative staphylococci isolated from blood of patients together with 41 strains of these bacteria isolated from the skin were typed by three different phage sets. Different percentages of phage typability by compared phage sets were found. Blood culture strains showed phage pattern Ph10/U14/U16 while other strains were of different phage patterns.

Bacteriophage Typing↗
PubMed2005

The two-component cell lysis genes holWMY and lysWMY of the Staphylococcus warneri M phage varphiWMY: cloning, sequencing, expression, and mutational analysis in Escherichia coli.

From the genome library of Staphylococcus warneri M, the two successive cell-lysis genes (holWMY and lytWMY) were cloned and characterized. The lytWMY gene encoded a protein (LysWMY), whose calculated molecular mass and pI were 54 kDa and 8.95, respectively. When overproduced in Escherichia coli, lysWMY directed a protein of 45 kDa (smaller than the predicted molecular mass), having N-terminal 13 residues identical with those predicted from DNA. Comparative analysis revealed that LysWMY significantly resembles the putative N-acetylmuramoyl-L-alanine amidases encoded by the staphylococcal phages phi11, 80 alpha, and Twort. Examination of modular organization of LysWMY identified three putative domains CHAP (for D-alanyl-glycyl endopeptidase), amidase (L-muramoyl-L-alanine amidase), and SH3 (cell wall recognition). Gene knockout analysis revealed that each of the two domains of CHAP and amidase was responsible for cell-lytic activity on a zymogram gel. Site-directed mutation of Cys29Ala, His92Ala, or Asn114Ala in the CHAP domain substantially reduced cell-lytic activity, suggesting that this Cys-His-Asn triad is crucial for the enzymatic function. On the other hand, the holWMY gene encoded a protein (HolWMY) with molecular mass and pI of 16 kDa and 4.36; this protein contained two potential transmembrane helices, resembling other predicted holins (a cytoplasmic membrane-disrupting protein) encoded by the S. aureus phage, phi11, 80 alpha, and Twort. Upon mitomycin C exposure of S. warneri M, a prophage (phiWMY) was induced and the virion was examined under electron microscopy. PCR amplification and sequencing revealed the presence of the holWMY-lysWMY genes in the phage genome.

Amidohydrolases↗
PubMed1961

Correlation between acid phosphatase and coagulase production or phage type of Staphylococcus aureus.

Pan, Yue-Liang (University of Michigan Medical School, Ann Arbor), and Harold J. Blumenthal. Correlation between acid phosphatase and coagulase production or phage type of Staphylococcus aureus. J. Bacteriol. 82:124-129. 1961.-A quantitative assay of acid phosphatase in 26 coagulase-negative and 73 coagulase-positive strains of Staphylococcus aureus was performed using an assay procedure modified to estimate extracellular, as well as the cellular, phosphatase. Some of the factors affecting the formation of acid phosphatase were studied. On the average, coagulase-negative strains produced 4.0 units and coagulase-positive strains 19.6 units although some negative strains produced more phosphatase than positive strains. More phosphatase was produced by strains of phage-group I than by strains of any other group.

Acid Phosphatase↗
PubMed1990

Changing phage pattern of Staphylococcus in pyoderma cases.

Staphylococcus aureus is a major pathogen for pyoderma in India. Phage pattern of bacteria gives valuable information in epidemiological studies of infection. Two hundred and two strains of Staphylococcus aureus isolated from pyoderma cases at Gorakhpur, were phage grouped and phage typed. It was found that 43.1 percent strains were not typable. Most common group was mixed phage group (23.8 percent) followed by phage group III (12.4 percent). Predominant phage types in mixed phage group was 84/81/85 and in phage group III was 84/85.

Bacteriophage Typing↗
PubMed1987

Molecular relatedness of Staphylococcus aureus typing phages measured by DNA hybridization and by high resolution thermal denaturation analysis.

Fifteen bacteriophages representative of the serological and lytic groups of the International Typing Set for Staphylococcus aureus were examined for genomic homology by DNA hybridization and by analysis of high resolution thermal denaturation profiles. Phages 11 and 80 alpha, not part of the set, were also examined. DNA homology measured by filter hybridization showed values ranging from near zero to 88 per cent in pair-wise comparisons. Cluster analysis of these data by standard numerical taxonomical methods yielded clusters which closely reflect the subdivision of the international set on the basis of serological reactions. High resolution thermal denaturation analysis yielded characteristic profiles for each phage DNA, with members closely related by hybridization analysis showing only minor differences. Quantitative analysis of the extent of overlap of these profiles generated relational values which were subjected to the same numerical taxonomic analysis as for the DNA hybridization data. The resultant dendrogram was qualitatively different only in minor respects from that derived from the hybridization analyses, but quantitatively homology was greater by 80 per cent or more for the DNAs which were least related according to the hybridization analyses. This upward shift in measured homology appears to reflect the similar base composition of the DNAs from these phages. Statistical comparison of the homology data obtained by the two methods showed them to be significantly correlated. These results indicate that the International Typing Set consists of phages which all appear to be related to a greater or lesser extent. If, as the history of the collection of these phages indicates, they are a random sample of aureophages, then this group of phages may represent a common genetic pool within which recombination, mutation, and genome rearrangement occur to generate unique individual phages.

DNA, Viral↗
PubMed2005

Sau42I, a BcgI-like restriction-modification system encoded by the Staphylococcus aureus quadruple-converting phage Phi42.

The serotype F phage Phi42 of Staphylococcus aureus is a triple-converting bacteriophage that encodes the staphylokinase gene (sak) and the enterotoxin A gene (entA). Lysogeny results in loss of expression of the chromosomal beta-haemolysin gene (hlb) (negative conversion), the expression of staphylokinase and enterotoxin A (positive conversion), and the acquisition of resistance to lysis by all 23 phages of the International Basic Set (IBS) of S. aureus typing phages. Until this study, the basis of Phi42 resistance to lysis by exogenous phages was unknown. The authors report here that phage Phi42 encodes a restriction-modification (R-M) system, termed Sau42I, adjacent to and in the same orientation to the phage integrase gene int. The genes encoding Sau42I were cloned and sequenced, and found to consist of two overlapping reading frames, ORF S (specificity) and ORF RM (restriction-modification), in the same orientation. The ORFs share a high degree of DNA and amino acid sequence homology with the previously characterized BcgI R-M system of Bacillus coagulans. Expression of the cloned Sau42I ORF S and ORF RM in S. aureus 80CR3 transformants from a plasmid vector conferred resistance to lysis by all 23 IBS phages. Similarly, transformants of S. aureus RN4220 harbouring recombinant plasmids containing both ORFs were resistant to lysis by the IBS typing phages. However, transformants harbouring plasmids encoding either ORF S or ORF RM were susceptible to lysis by the IBS phages, and they had the same phage-susceptibility pattern as the respective parental isolates. In vitro analysis of crude and partially purified extracts of S. aureus transformants harbouring both the Phi42 ORF S and ORF RM genes indicated that Sau42I has endonuclease activity and requires co-factors Mg(2+) and S-adenosylmethionine in order to function, and activity is optimized at pH 8, although the precise recognition sequence has yet to be determined. The findings of this study confirm that Phi42 is a quadruple-converting phage, believed to be the first described for S. aureus, and show that it encodes a novel R-M system termed Sau42I.

Amino Acid Sequence↗
PubMed1997

International quality control of phage typing of Staphylococcus aureus. International Union of Microbial Societies Subcommittee.

A questionnaire was sent to the 48 national typing centres for Staphylococcus aureus and 31 replies were received. Methods of phage typing varied and molecular methods were not universally available, although pulsed-field gel electrophoresis was offered by 13 centres. Results for a quality control phage typing exercise were received from 25 centres. Increased standardisation of methods and definitions are indicated. Differences from the consensus patterns were mainly due to typing at an inappropriate dilution of phage, but five strains caused difficulties in many centres. Overall reproducibility was good. Phage typing remains a cost-effective method for epidemiological studies, particularly on a large scale. The strains selected for the quality control exercise included many strains suitable for controlling molecular methods as well as testing phage typing. Molecular methods help in the validation of the conclusions which may be drawn from phage typing.

Bacterial Proteins↗