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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↗

Bacteriophage typing of Shigella sonnei.

A bacteriophage-typing schema was developed for differentiating strains of Shigella sonnei. Sixty-seven bacteriophages were obtained from other collections, and 36 bacteriophages were isolated from sewage. From these 103 bacteriophages, a provisional set of 12 was chosen by computer analysis as being the most sensitive in differentiating strains of S. sonnei isolated in the United States. The provisional schema was used to type 265 strains from different geographical areas. It divided them into 87 different lysis patterns, and all 265 strains were typable. Smooth and rough colonial variants of the same strain had different lysis patterns, so the technique was standardized to type rough colonies only. Reproducibility was difficult to obtain until all conditions were carefully standardized. Changes in results were noted even on different lot numbers of Trypticase soy agar, which was defined as the standard medium. So that the medium would not be a variable, 100 pounds (ca 453.5 kg) of the same lot number was purchased. Bacteriophage typing was very useful in differentiating strains, and work should continue on establishing a standarized schema.

Bacteriophage Typing↗

Bacteriophage typing of coagulase-negative staphylococci.

Cultures comprising the 10 species of coagulase-negative staphylococci proposed by Kloos and Schleifer (J. Clin. Microbiol. 1:82--88, 1975) were typed with bacteriophages isolated from Staphylococcus epidermidis. Although only 10.5% were typable, 50% of those identified as S. epidermidis were typed. Cultures from patients with middle ear infections were also classified by this system and phage typed.

Bacteriophage Typing↗

Bacteriophage typing scheme for Salmonella infantis.

A bacteriophage typing system is described for Salmonella infantis. Nine phages were selected, of which three were isolated from sewage and six from human feces. All except 7 of the 546 strains collected between 1974 and 1978 could be classified into 23 different phage types. The five most common phage types comprised 26, 13, 9, 9, and 9% of all strains, respectively. Strains from humans, animals, food, and water isolated during nine episodes, or from given patients at different intervals of time, belonged to the same phage type.

Bacteriophage Typing↗

[Bacteriophage types represented by lactose-fermenting Salmonella agona strains].

Bacteriophage typing was performed on 1911 S. agona, lactose-fermenting strains. These strains were isolated from hospitalised newborns and neonates patients. Out of 1911 strains 98.8% were typable by means of phage set prepared for strains differentiation of Salmonella agona showing typical biochemical properties. It was shown that in 16 provinces from which the strains were obtained in 1983-1985 type V (49.5%) and type XI (25.4%) prevailed. Subtypes VA and VB were distinguished within type V. Altogether 20.3% of strains were classified as belonging to these subtypes. Their lytic reaction was weaker with phages 3, 4, and 9 with the characteristic range of phage type V strains. Among tested strains types I, XIII, and XVI were also represented composing 2, 6, 0, 9, and 0.3% of total number of strains respectively. 1.5% of strains were nontypable and 0.2% showed lytic reactions different from that included in up to now used scheme of typing. It can be concluded that lactose-fermenting S. agona strains show susceptibility to lowered number of phages than typical for Salmonella species strains. It seems that differentiation of this atypical biochemical variant of S. agona with, the use of phage set used up to now may be also usefull in practice as it is the case in respect to strains with typical biochemical properties.

Bacteriophage Typing↗

[Distribution of bacteriophage types of Mycobacterium tuberculosis in France].

The bacteriophage typing of M. tuberculosis enables separation of the strains of the species according to their sensitivity to certain bacteriophages. A relationship has been observed between the geographical origin of patients and the distribution of the phage types of their strains. This study related to a sample of 450 strains isolated between 1978 and 1987 in patients of French origin coming from different regions in France. An analysis of the data provided by the study indicates that the distribution of the 5 phage types, A, AX, I, B, and C is relatively homogenous in the different regions with a predominance of the phage type A and B. The mean percentage of the lysotypes A, AX, I, B and C in the country as a whole are 43%, 15%, 7%, 30% and 5% respectively. Only in region 3 (Haute and Basse Normandie), 8 (Provence-Côte-d'Azur) and 9 (Rhône-Alpes) is there any perceptible deviation from this distribution. This study also underlines the contribution that can be provided by this type of information in the epidemiology of tuberculosis.

Bacteriophage Typing↗

Structural proteins and DNA characteristics of 14 Listeria typing bacteriophages.

The major structural proteins of 13 temperate and one virulent Listeria typing bacteriophages were analysed and compared using isoelectric focusing in immobilized pH gradients (IPG), ultrathin-layer two-dimensional electrophoresis, amino acid analysis and N-terminal amino acid sequences of selected proteins. Isoelectric points for major capsid and tail proteins of the 12 members of the siphoviridae family included in this study ranged from 4.70 to 5.92, whereas one of the two myoviridae investigated (B054) showed structural proteins in the 6.1 to 6.3 range. In comparison to protein profiles from one-dimensional SDS gels, the IPG technique gave better resolution and improved discrimination of phage proteins. Combination of this technique and SDS gel electrophoresis made it possible to correlate M(r) and isoelectric points of major structural proteins. Tail polypeptides of all siphoviridae are smaller and, with one exception, more acidic than their corresponding capsid counterparts. We also determined the amino acid composition of capsid and tail proteins. When compared with an average protein, they were found to be fairly rich in acidic and short-chain hydrophobic amino acids, as well as in lysine. In addition, the N-terminal amino acid sequences of major capsid and tail proteins of four representative listeriaphages were compared. The base composition of listeriaphage DNAs was between 37% and 39% G + C, reflecting that of their bacterial hosts. Each phage had a distinct restriction endonuclease pattern, and genome sizes ranged from 35 to 116 kb. DNA-DNA hybridization permitted the identification of five DNA homology groups. The two myoviruses studied (A511 and B054) showed no DNA homology to other phages, confirming their unique nature. The 12 siphoviruses were classified into three DNA homology groups with little cross-homology. Furthermore, phage A006 was found to share little DNA homology with the other investigated members of species 2671. Therefore, a new species (A006) is proposed. With respect to phage classification and taxonomy, a good correlation between the various approaches was observed, mostly corresponding to particle morphology.

Amino Acid Sequence↗

A catalogue of T4-type bacteriophages.

The T4-type of bacteriophages is broadly defined on the basis of particle morphology. It occurs in enterobacteria (125 representatives), acinetobacters, aeromonads, pseudomonads, and vibrios (16 isolates). In addition, 18 apparently unrelated phages with prolate heads and contractile tails are found in a wide range of bacteria. A descriptive catalogue of these phages is presented. The T4-type probably originated in precursors of enterobacteria.

Bacteriophage T4↗

Stability of bacteriophage type of Mycobacterium tuberculosis: absence of variation caused by experimental chemotherapy in mice and analysis of spontaneous variation.

The bacteriophage typing of 54 strains isolated from the tissues of mice infected with the strain H37Rv of Mycobacterium tuberculosis and treated with isonazid and rifampin showed that the bacteriophage type did not change. The fluctuation analysis of populations of the tubercle bacilli demonstrated that the rate of mutation from sensitivity to resistance in respect to the bacteriophage DS6A was less than 1.3 X 10(-8) mutations per bacterium per generation.

Analysis of Variance↗

Nosocomial transmission of bacteriophage type 7/11/12 group B streptococci in a special care nursery.

Two premature infants in a special care nursery acquired late-onset group B streptococcal (GBS) sepsis within a 24-hour period. The infecting strains were serotype III organisms with bacteriophage type 7/11/12. Cultures of the mothers of the two affected infants were negative for GBS, implying nosocomial acquisition of infection. Although 32% of nursery personnel had mucosal carriage of GBS, none of the seven isolates of GBS type III was the same bacteriophage type as the two infecting strains. Of the other infants hospitalized in the nursery, five were asymptomatically colonized with GBS. These infants were in bassinets adjacent to the affected infants; all five of their isolates were identical to the two infecting strains. We conclude that infant-to-infant transmission may result in nosocomial late-onset GBS septicemia.

Bacteriophage Typing↗

Development of a bacteriophage typing system for Campylobacter jejuni and Campylobacter coli.

A bacteriophage typing system for Campylobacter jejuni and Campylobacter coli was developed with phages isolated from poultry feces. Data for phage selection were generated from a set of isolates of C. jejuni and C. coli from humans in Illinois. Selection of 14 phages from the 47 phages available was assisted by determination of the Sneath-Jaccard similarity coefficients and subsequent unweighted pair-group arithmetic averaging cluster analysis. The typing set was reproducible and stable in the 255 isolates from Illinois. Of these isolates, 94.5% were typable, with 46% represented by the four most common phage patterns. In a set of 51 isolates from humans outside of Illinois, 88.1% of the C. jejuni isolates were typable. Phage typing for C. jejuni and C. coli has excellent epidemiologic potential and should serve as a useful adjunct or alternative to serotyping systems in current use.

Animals↗

[Epidemiological analysis of Salmonella enteritidis isolates using pulsed-field gel electrophoresis and bacteriophage typing over the period of April 2000 to March 2003 in Gifu Prefecture].

We examined a total of 151 Salmonella enterica serovar Enteritidis strains isolated in Gifu Prefecture during the period from April 2000 to March 2003 by using bacteriophage typing and pulsed-field gel electrophoresis (PFGE). Bacteriophage typing classified them into twelve phage types (PT) and RDNC (reacted but did not conform). The predominant phage type was PT47 (34.4%) followed by PT1 (21.9%), PT4 (16.6%) and RDNC (11.3%). XbaI- and BlnI-digested PFGE analyses identified 17 and 44 PFGE patterns, respectively, indicating that PFGE with BlnI had more discriminating power than that with XbaI. Combination of the phage types and PFGE types of BlnI could make 53 subtypes. Some isolates with the same phage type were subdivided into different PFGE types, but those with PT47 were not. PT47 isolates were derived from sporadic patients with gastroenteritis, food poisoning outbreaks and healthy carriers through the years. This suggests that PT47 is highly clonal and disseminates over our prefecture.

Bacteriophage Typing↗