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Diphthericin types, bacteriophage types and serotypes of Corynebacterium diphtheriae strains isolated in Australia.

A dipthericin typing scheme has been constructed using 441 strains of Corynebacterium diptheriae isolated in eastern Australia from 1962 to 1971. Ten types have been distinguished using seven strains of C. diphtheriae and two strains of C. belfanti as indicators of the diphthericins produced by the newly isolated strains. Strains grouped into types L2, L3 and L3a were found only in Melbourne and types L1 and L4 were predominant in Sydney. Type L5 strains were isolated intermittently throughout the period of study and were found in all eastern states. Numerical analysis of the characteristics of the strains suggests that associations exist between, on the one hand, diphthericin type and, on the other hand, bacteriophage type, serotype and biochemical activity.

Agglutination Tests↗

Failure of bacteriophage typing to detect an inter-hospital outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb subsequently identified by random amplification of polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE).

OBJECTIVE: To establish the extent of inter-hospital spread of methicillin-resistant Staphylococcus aureus (MRSA) in Zagreb and to determine the most suitable method for typing local strains. METHODS: We analyzed a collection of 33 MRSA isolates from three Zagreb hospitals together with five unrelated British MRSA isolates by antibiogram typing, bacteriophage typing, randomly amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE) after digestion with Smal restriction endonuclease. Bacteriophage typing was done with the international set of S. aureus typing phages. RAPD and PFGE profiles were analyzed visually and by using the 'GelCompar' computer program. RESULTS: Antibiogram typing provided eight profiles. Thirty (91%) of the 33 Croatian strains of MRSA were non-typable by phage typing. Visual analysis of RAPD products identified six, and visual analysis of PFGE fragments nine, distinct profiles. Computer analysis of RAPD data separated British isolates from the Croatian ones, but did not cluster the visually determined RAPD types. PFGE computer analysis separated British isolates and clustered isolates in concordance with visual interpretation. Thirty-one of the 38 isolates (82%) were visually grouped in the same clusters by both molecular methods. The dominant strain was present in each of the three hospitals. CONCLUSIONS: Bacteriophage typing was unhelpful for the analysis of Croatian MRSA, since most strains were untypable with the international set of bacteriophages. RAPD and PFGE were more successful in typing the organisms and showed evidence of inter-hospital spread of one predominant MRSA strain in all three Zagreb hospitals. Thus RAPD and PFGE proved to be a useful aid in elucidating the epidemiology of MRSA infection in Zagreb hospitals and should be established in Croatia for typing MRSA.

Journal Article↗

Bacteriophage typing of Salmonellae. II. New bacteriophage typing scheme.

A phage-typing technique for salmonellae is described. One battery of phages was used to type three serotypes of Salmonella, namely, S. typhimurium, S.typhimurium var. copenhagen, and S. heidelberg. In all, 443 S. heidelberg cultures were typed into 22 phage types, 185 S. typhimurium cultures into 35 phage type, and 92 S. typhimurium var. copenhagen cultures into 26 phage types. The stability of the phage types was established by retyping 168 cultures belonging to all three serotypes. The epidemiological significance of the phage types demonstrated was evaluated by comparing phage types obtained from the University of Minnesota and those from the National Animal Disease Laboratory. Further investigation of the S. heidelberg phage types has shown that the cultures represented repeated isolates from the same birds or from a group of birds in the same flock.

Animals↗

Subtyping of methicillin-resistant Staphylococcus aureus isolates from the North-West of England: a comparison of standardised pulsed-field gel electrophoresis with bacteriophage typing including an inter-laboratory reproducibility study.

Bacteriophage typing is currently the recognised methodology for the typing of methicillin-resistant Staphylococcus aureus (MRSA) in the UK. Bacteriophage typing is less discriminatory and does not type all isolates compared with some molecular methods for typing MRSA. Chromosomal genotyping by pulsed-field gel electrophoresis (PFGE) is increasingly recognised as an improved method for typing MRSA, providing increased discrimination and typability. In this study the results of a comparison of bacteriophage typing and PFGE typing and subtyping are presented for a large collection of isolates from the North-West of England. Isolates belonging to the most frequently isolated epidemic methicillin-resistant Staphylococcus aureus (EMRSA) bacteriophage types 15 and 16 were typed by PFGE with further discrimination of common PFGE types possible into a number of subtypes. These results for a large collection of isolates demonstrate the improved typing of MRSA with PFGE. The widespread acceptance of PFGE for typing MRSA isolates has been hampered by the lack of standardised methodologies. Recently, a standardised PFGE strain typing system, known as the GenePath system has become available. The results of an inter-laboratory comparison of PFGE typing for a collection of isolates demonstrated good reproducibility with this system.

Bacterial Typing Techniques↗

Reproducibility of staphylococcal bacteriophage types by use of different concentrations of bacteriophage.

A comparative investigation was carried out on the reproducibility of staphylococcal bacteriophage types at 100 routine test dilution (RTD) and at the conventional concentrations, RTD and 1,000 RTD. The comparison includes multiple typing of laboratory strains and duplicate isolates from hospital patients. No significant difference was found in the reproducibility of the types at the different concentrations. A single concentration corresponding to 100 RTD is recommended for routine use in phage typing as a possible substitute for typing with two different concentrations.

Bacteriophage Typing↗

Bacteriophage typing of Corynebacterium diphtheriae.

Bacteriophage types of over 3,000 strains of Corynebacterium diphtheriae isolated in Canada have been determined. The typing scheme used involved the use of nine phages. Results indicated that phage types correlate with biotypes to a large degree. Corynecin types were also determined for a limited number of cultures, and results indicated that the indicator strains presently available are unsuitable for the typing of C. diphtheriae strains isolated in North America. The distribution of phage types is similar throughout Canada, and the types present correspond to types reported from other countries. Phage typing could be of value in the study of circumscribed outbreaks and in epidemiological surveillance of types over long periods.

Bacterial Toxins↗

Pulsed-field gel electrophoresis as a replacement for bacteriophage typing of Staphylococcus aureus.

Bacteriophage typing (BT) (World Health Organization method) has been used at the Centers for Disease Control and Prevention for over 30 years to type isolates of Staphylococcus aureus. Since studies have shown that BT patterns have poor reproducibility and because BT fails to type a high percentage (15 to 20%) of isolates, the Centers for Disease Control and Prevention has converted from using BT to using pulsed-field gel electrophoresis (PFGE) for strain typing S. aureus. We compared the results of BT with results of PFGE for typing 300 isolates of S. aureus, including strains from several well-characterized outbreaks. Ninety-six isolates were BT group I, 19 were group II, 82 were group III, 7 were group V, and 96 were nontypeable. PFGE identified subgroups within each phage group and thus was more discriminating than BT, which identified no subgroups. PFGE was able to type all isolates and distinguish related from unrelated strains of S. aureus. Our modified, standardized PFGE methodology should enable typing laboratories to obtain rapid, reliable results in 3 to 4 days when starting with an isolated colony on agar media.

Bacteriophage Typing↗

Bacteriophage typing of clinically isolated Serratia marcescens.

A bacteriophage-typing scheme for the differentiation and classification of clinically isolated strains of Serratia marcescens was developed. Thirty-four Serratia bacteriophages were isolated from sewage and used to type 185 of 204 isolates (90.6%) of S. marcescens into 23 bacteriophage groups representing 71 types. Different bacteriophage types occurred at different intervals, suggesting that particular strains of S. marcescens are found at certain times. A correlation was found between inositol fermentation and bacteriophage type and between susceptibility to carbenicillin and bacteriophage type. However, there was no relationship between source of isolate and bacteriophage type. Bacteriophage typing of S. marcescens should provide a system which will aid in determining the origin of nosocomial Serratia infections.

Bacteriophage Typing↗

Bacteriophage typing of Listeria species.

A bacteriophage typing scheme for differentiating Listeria isolates from dairy products and various other foodstuffs was developed. Sixteen selected phages isolated from both environmental sources and lysogenic strains were used for typing and, according to their lytic spectra, divided into four groups. Thus far, 41 distinct patterns of lysis were seen when this set was used in typing 57 defined reference strains, representing all five confirmed species and 16 serotypes in addition to 454 Listeria isolates of primarily foodborne origin. Overall, typability was 84.5%; i.e., a strain was lysed by at least one phage at 100x routine test dilution. Strains belonging to serovar 3 were mostly resistant to lysis by the phages employed. The results were highly reproducible, as determined in retyping trials several weeks later. Some phages isolated from environmental sources showed a wider lytic spectrum than did those isolated from lysogenic strains. In accordance with this, the phages were found in different clusters within a computer-generated linkage map. Species specificity and serovar specificity of the lytic reaction were not found. None of the phages was able to lyse strains of Listeria grayi, Listeria murrayi or Jonesia denitrificans. This phage typing system may provide important information for a means of recognizing and eliminating sources of contamination by Listeria spp. within dairy plant equipment.

Bacteriophage Typing↗

[Current methods of intraspecies typing of Shigella sonnei. 3. Bacteriophage typing].

Data are presented concerning the study of the phage type of Sh. sonnei isolated in various regions of the USSR. The strains isolated belonged to 64 phage types corresponding to the Hammarström's scheme. A variety of Shigella sonnei phage types both in one and in different territories of the country provided future prospects for successful use of phage typing for the purpose of epidemiological supervision and analysis of dysentery morbidity.

Bacteriological Techniques↗

[Characterization of B-type bacteriophages adsorbed on Pseudomonas aeruginosa pili].

Pilus-dependent B-morphotype bacteriophages isolated from various sources were studied. The adsorption of phages on Pseudomonas aeruginosa pili was proven. Electron-microscopic examination of the morphology of phages was carried out, and the adsorption properties were partially described. It was suggested that adsorption apparatuses of different pilus-dependent B-phages are alike with respect to structure and function.

Adsorption↗

Development and epidemiological applications of a bacteriophage typing system for typing Pasteurella multocida.

A bacteriophage typing system was developed for typing toxigenic and nontoxigenic strains of Pasteurella multocida. A phage set of 24 phages with different lytic spectra was isolated after mitomycin treatment of P. multocida strains, isolated mainly from pigs from herds with atrophic rhinitis. On a test set of 97 different strains isolated from pigs, these 24 phages were able to type 87% of the strains. The 97 test strains could be subdivided into 31 different types by reaction with the 24 phages. The reproducibility after subculture and storage of the strains was good (95%). Phage typing of 217 toxigenic P. multocida field isolates from 37 pig herds predominately with clinically atrophic rhinitis resulted in 18 different phage types and an overall typability of 68%. Of 24 herds from which more than three isolates of toxigenic P. multocida were obtained, a single phage type was demonstrated in 5 herds, while in 9 herds a single phage type represented at least half of the isolates. The phage types in the remaining 10 herds revealed no dominating phage type. The phage typing system described appears to be a valuable epidemiological tool for studying the spread of P. multocida.

Animals↗

Base ratio and deoxyribonucleic acid homology studies of six Staphylococcus aureus typing bacteriophages.

Genetic relatedness among Staphylococcus aureus typing bacteriophages 80, 47, 81, 71, 77, and 187 was investigated by using base ratio determinations and deoxyribonucleic acid (DNA)-DNA hybridization. Guanine/cytosine (G/C) content, as determined by thermal denaturation and chromatographic analysis of the purines released by acid hydrolysis of the DNA, was between 31 and 36%. No pattern correlating G/C content with serological or lytic group was discernible. DNA-DNA hybridization studies indicated high degrees of homology (43% or more) among the genomes of phages in the same serological group. Less homology (29% or less) was observed between the genomes of phages belonging to different serological groups. These findings implied a positive correlation between serological and genetic relatedness.

Bacteriophage Typing↗

Differentiation of Proteus mirabilis by bacteriophage typing and the Dienes reaction.

A provisional typing schema based on sensitivity to 23 bacteriophages has been established for Proteus mirabilis. Seventy-three bacteriophages were isolated on strains of P. mirabilis (64), P. vulgaris (1), P. morganii (7), and P. rettgeri (1), but those isolated on P. mirabilis were the most useful in differentiating other strains of . mirabilis. From the 73 phages studied, the best 23 were chosen by computer analysis for the provisional system, which was then used to study P. mirabilis infections in a 500-bed general hospital. All patient isolates for 19 months were saved and then compared by bacteriophage typing and the Dienes reaction in a retrospective study. There was evidence for only three instances of cross-infection or -colonization during this time. Bacteriophage typing was very sensitive in differentiating strains, since 200 strains were differentiated into 113 different lysis patterns and 94% were typable. The Dienes reaction was useful at times but often gave reactions that were difficult to read or that changed when the tests were repeated. The bacteriophages described by Schmidt and Jeffries were also evaluated and proved useful in combination with ours. The value of bacteriophage typing was clearly established, and work toward a standardized schema for P. mirabilis should continue.

Bacteriophage Typing↗