Effective proliferation of low level Legionella pneumophila serogroup 1 cells using coculture procedure with Acanthamoeba castellanii.

The multiplications of low level Legionella pneumophila serogroup 1 cells by the coculture procedure with Acanthamoeba castellanii were tested in five strains. The cells in all strains proliferated effectively for isolating. This procedure might be a useful means of improving the successful isolation from environmental and clinical specimens of low level Legionella cells, and pursuing the source of infection.

Genomic analysis of a multidrug-resistant strain of enterohaemorrhagic Escherichia coli O157:H7 causing a family outbreak in Japan.

A family outbreak of enterohaemorrhagic Escherichia coli (EHEC) O157:H7 infection occurred in October 2003 in the Hiroshima prefecture, Japan. Four isolates of EHEC O157:H7, 03064, 03065, 03066 and 03067, were recovered from a 1-year-old daughter, mother, father and 3-year-old daughter, respectively. All EHEC O157:H7 isolates were positive for Stx1 and Stx2 Shiga toxins. Surprisingly, DNA fingerprinting profiles obtained by PFGE showed that the first isolate, 03064, had unique XbaI and BlnI profiles that differed from the other three isolates. Also, plasmid analysis results revealed that isolate 03064 contained an extra plasmid larger than the classic large plasmid of EHEC O157, pO157 (93.6 kb). This new plasmid was named pMDR157. Furthermore, isolate 03064 showed a multidrug-resistance (MDR) phenotype against streptomycin, spectinomycin, co-trimoxazole (trimethoprim/sulfamethoxazole), ampicillin and tetracycline; the other isolates were completely sensitive to these antibiotics. Molecular analysis of the MDR phenotype in this unique strain revealed the presence of a class 1 integron containing two gene cassettes: a dihydrofolate reductase type 1 gene (dfrI), which confers resistance to trimethoprim, and an aminoglycoside adenyltransferase gene (aadA1), which confers resistance to streptomycin and spectinomycin. Southern blot hybridization showed that the class 1 integron was located in the extra plasmid, pMDR157. The ampicillin resistance was found to be due to the presence of the TEM-1-type beta-lactamase gene. The MDR phenotype was transferred successfully to E. coli HB101 by conjugation, indicating that both the class 1 integron and the TEM-1 beta-lactamase were located on the conjugative transferable plasmid, pMDR157. To the authors' knowledge, this is the first report of the identification of a beta-lactamase gene in EHEC O157.

[Epidemiological and bacteriological investigation of enterohemorrhagic Escherichia coli infection in the Chugoku-Shikoku area].

We investigated the occurring tendency of enterohemorrhagic Escherichia coli (EHEC) infection in the prefectural and municipal public health institutes in the Chugoku-Shikoku area from 1996 to 1999, and the bacteriological characteristics of EHEC isolated from these cases. Consequently, epidemiological analysis of the EHEC infection in this district was performed. 22 outbreaks in the various facilities showed the tendency occurred in infants and aged groups, and the serotypes of EHEC isolated from these outbreaks were O26, O111 etc. as well as O157. In 4 cases, EHEC were isolated from specimens of buckwheat noodles, salad, sand box, and goat feces, and these were determined as the source of infection. In 898 sporadic cases, including familial infection, the EHEC isolates were classified into 24 serotypes, and the genotypes of EHEC O157:H7 isolates by pulsed-field gel electrophoresis (PFGE) also varied. Moreover, since many asymptomatic carriers were detected in the adult group with familial infection, the existence of healthy carriers is as important as the source of infection. The drug-resistance test of EHEC isolates showed that 24% of the 924 isolates were resistant to drugs.