[Bacteriophage typing of Staphylococcus aureus and study of the sensitivity to antibiotics].
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We have recently cloned three DNA fragments (In-2.6, In-1.0, and In-0.6) of the non-cultured, bacterial-like organism (BLO) associated with citrus greening disease. Nucleotide sequence determination has shown that fragment In-2.6 is part of the rplKAJL-rpoBC gene cluster, a well-known operon in eubacteria. The DNA fragment upstream of and partially overlapping with In-2.6 could be isolated and was shown to be the nusG gene. In Escherichia coli, nusG is also immediately upstream of rplKAJL-rpoBC. Fragment In-1.0 carries the gene for a bacteriophage type DNA polymerase. Fragment In-0.6 could not be identified. When In-2.6 was used, at high stringency, as a probe to detect greening BLO strains in infected plants, hybridization was obtained with all Asian strains tested, but not with the African strain examined. At lower stringencies, In-2.6 was able to detect also the African strain. The implications of these results in the taxonomical position of the greening BLO are discussed.
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The typing phages set of Ward et al. was used to type a total of 517 strains of Salmonella Enteritidis isolated in Poland in 1986-1995. According to the Ward et al. scheme, 56.5% of the strains tested were assigned to 14 different phage types. Phage types 8, 4, 1 and 4a were placed first, second, third and fourth, respectively. They were dominated both in the outbreak isolates and in the isolates from the other sources. Ten phage types were represented by single strains. Other strains reacted with phages without showing any of the designated phage type (37.1%) or were untypable (6.4%). The Ward et al. scheme seems to demonstrate not enough high degree of strain discrimination for Salmonella Enteritidis isolates in Poland. It seems that the Ward et al. scheme is not enough useful for the epidemiological investigations of Salmonella Enteritidis in Poland.
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Seventy-six phages active against Enterobacter cloacae were isolated from sewage and other sources. They were tested at RTD on 92 selected strains of E. cloacae and their lytic reactions were used to select phages for a typing set. Numerical analysis by the Jaccard coefficient was used to assess the similarity between the phages. A computer-based test selection procedure selected sub-sets of phages to discriminate between all 92 E. cloacae strains and within the most frequent serological groups. A subjective analysis of the candidate phages based on similarity, clarity of plaque and frequency of lysis was combined with the computer selected sub-sets to produce a final set of 25 phages that gave a good theoretical discrimination of clinical isolates of E. cloacae.
We examined a number of bacteriophages with T4-type morphology that propagate in different genera of enterobacteria, Aeromonas, Burkholderia, and Vibrio. Most of these phages had a prolate icosahedral head, a contractile tail, and a genome size that was similar to that of T4. A few of them had more elongated heads and larger genomes. All these phages are phylogenetically related, since they each had sequences homologous to the capsid gene (gene 23), tail sheath gene (gene 18), and tail tube gene (gene 19) of T4. On the basis of the sequence comparison of their virion genes, the T4-type phages can be classified into three subgroups with increasing divergence from T4: the T-evens, pseudoT-evens, and schizoT-evens. In general, the phages that infect closely related host species have virion genes that are phylogenetically closer to each other than those of phages that infect distantly related hosts. However, some of the phages appear to be chimeras, indicating that, at least occasionally, some genetic shuffling has occurred between the different T4-type subgroups. The compilation of a number of gene 23 sequences reveals a pattern of conserved motifs separated by sequences that differ in the T4-type subgroups. Such variable patches in the gene 23 sequences may determine the size of the virion head and consequently the viral genome length. This sequence analysis provides molecular evidence that phages related to T4 are widespread in the biosphere and diverged from a common ancestor in acquiring the ability to infect different host bacteria and to occupy new ecological niches.
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Species, biotypes, and phage types were determined for 120 Acinetobacter strains from clinical or environmental sources or from culture collections. These characteristics were compared with cell envelope protein profiles obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in previous studies. A considerable heterogeneity of species and types was observed by use of the various methods, in particular among strains from different sources. Acinetobacter baumannii was the most commonly found species in isolates from clinical sources, followed by Acinetobacter species 3. Nine biotypes were observed among A. baumannii strains. Further differentiation within most species and biotypes was achieved by protein profile typing and, to some extent, phage typing. Of 120 strains, 49 (41%) were not typeable by phages. Consistent results for the various classification methods were obtained for strains from common sources. Biotyping seemed an appropriate method for the screening of strains, whereas protein profile and phage typing could serve as additional methods to establish the identity or nonidentity of strains. Results of this study suggest that the combination of the typing methods is useful in epidemiological studies.