Salmonella enterica serovar Typhi in Japan, 2001-2006: emergence of high-level fluoroquinolone-resistant strains.

The phage types and antimicrobial susceptibilities of 226 isolates of Salmonella enterica serovar Typhi from imported cases in Japan between 2001 and 2006 were investigated. Most (93.8%) had travelled to Asian countries, particularly South East Asia. Twenty-one phage types were identified with E1 (30.5%), UVS (15.9%) and B1 (9.3%) being the most common. The frequency of multidrug-resistant strains reached 37.0% in 2006 with phage types E1 and E9 predominating. Almost half (48.2%) of the isolates were resistant to nalidixic acid and two isolates displayed high-level fluoroquinolone resistance. Three mutations, two in gyrA and one in parC, were identified in both isolates.

Species-identification dots: a potent tool for developing genome microbiology.

Identification of species has long been done by phenotype-based methodologies. Recently, genotype-based species identification has been shown to be possible by way of Genome profiling, which is based on a temperature gradient gel electrophoresis (TGGE) analysis of random PCR products. However, the results, though sufficient in information, provided by genome profiling were complicated and difficult to deal with objectively. To cope with this, a technology of utilizing species identification dots (spiddos), which corresponds to structural transition points of DNAs, was introduced. Pattern similarity score (PaSS), derived from spiddos, was shown to be usable for quantitatively measuring the closeness between genomes. This was demonstrated with the experiments applied to the genomes of Escherichia coli O157:H7 (19 strains). The same genomes were also examined by sequencing and RFLP methods in order to compare the effectiveness of these three methods. As a result, the spiddos method was shown to give reasonable results and to be the most advantageous for measuring the closeness between species in general. This means that spiddos is pushing the heavy gate open for genome microbiology.

[Diversity of DNA sequences among enterohemorrhagic Escherichia coli O157:H7 detected by PCR-based DNA fingerprinting].

A random amplified polymorphic DNA(RAPD) fingerprinting method has been developed to differentiate Enterohemorrhagic Escherichia coli(EHEC) O157:H7 isolates. This method uses an oligonucleotide of arbitrarily chosen sequence to prime DNA synthesis from pairs of sites to which it is matched or practically matched and results in strain-specific arrays of DNA products. By the RAPD analysis using A07(5'-TGCCTCGCACCA-3'), EHEC strains tested in this study were found to be divided into 5 groups. The RAPD arrays among 5EHEC clinical isolates from a single outbreak were identical from each other, and different from other origin. The data indicate that the RAPD method is feasible for investigating the source of an outbreak associated with EHEC O157:H7 infection.

Diversity of DNA sequences among Vibrio cholerae O139 Bengal detected by PCR-based DNA fingerprinting.

Vibrio cholerae O139, a causative agent of a large epidemic of cholera-like illness, has suddenly emerged and spread widely over several months. To investigate the characteristics unique to O139, traditional typing techniques for V. cholerae, such as biochemical characteristics, antibiotic susceptibility and detection of toxin production, were performed, with the result that 145 O139 strains, except for two O139 strains isolated from Argentina and Germany, were indistinguishable from O1 strains. Thus, in order to clarify the genetical relatedness among O139 strains, and between O139 and O1 strains, the RAPD (random amplified polymorphic DNA) DNA fingerprinting method was undertaken. Although the RAPD arrays in five O139 isolates from Vellore with one arbitrary primer were slightly different from the other O139 strains, the RAPD patterns of the 145 forty-five O139 strains except for two O139 strains from Argentina and Germany were quite similar to each other, but were different from those of O1 strains, indicating that those O139 epidemic strains are closely related to each other regardless of their place of isolation. Furthermore, the RAPD patterns of the O139 strains resembled those of E1 Tor strains rather than classical strain, and a small change in the RAPD pattern of O139 strains occurred during subculture for 200 generations. These results taken together suggested that O139 V. cholerae have emerged from a common origin associated with the E1 Tor strain.

[Biological characters of enterohemorrhagic Escherichia coli isolates from diarrhea patients in Saitama (1990-1992)].

A total of 16 strains of Enterohemorrhagic Escherichia coli (EHEC) isolated from diarrea patients in Saitama from 1990 to 1992 were tested for their serotype, verotoxin production, biochemical characteristics, antibiotics sensitivity and plasmid profiles. By serotype analysis, 14 strains from two outbreaks and 12 sporadic cases were classified as type O157:H7, one as O111:H-(not motility) and one as O128:H2. Typing of verotoxin by gene analysis using Polymerase Chain Reaction (PCR) showed that 9 of O157:H7 strains including two cases from outbreaks and O128:H2 have VT1 and VT2 genes, other O157:H7 have the VT2 gene and O111:H-has only the VT1 gene. Biochemical characteristic analysis indicated two strains of O157:H7 type from outbreaks were biotype II and the rest of O157:H7 were biotype I. One of the O157:H7 strain from a sporadic case showed positive for urease production. According to sensitivity tests against antibiotics, out of the O157:H7 group, one strain was resistant against ABPC, one against SM and two strains resistant to SM-TC. For plasmid profiles, all strains had 94 Kb plasmids and several smaller sizes of plasmids. But 5 strains of O157:H7 had 94 Kb plasmid only.

[The first report of traveler's diarrhea associated with a newly described toxigenic Vibrio cholerae O139 strain in Japan].

A newly described Vibrio cholerae O139 was isolated from a patient who had traveled in India on April 1993. The patient experienced 5 to 6 watery diarrhea per day after he returned to Japan. The isolated strain registered as K111 did not agglutinate with O1-O138 antiserum and agglutinated with O139 antiserum. This strain resembled V. cholerae O1 strain in biochemical characters and had ctx and zot, although was resistant to the vibrio static agent O/129. This is the first report of cholera-like illness by the newly described toxigenic V. cholerae O139 strain in Japan.