Taxonomy of Australian clinical isolates of the genus Photorhabdus and proposal of Photorhabdus asymbiotica subsp. asymbiotica subsp. nov. and P. asymbiotica subsp. australis subsp. nov.

The relationship of Photorhabdus isolates that were cultured from human clinical specimens in Australia to Photorhabdus asymbiotica isolates from human clinical specimens in the USA and to species of the genus Photorhabdus that are associated symbiotically with entomopathogenic nematodes was evaluated. A polyphasic approach that involved DNA-DNA hybridization, phylogenetic analyses of 16S rRNA and gyrB gene sequences and phenotypic characterization was adopted. These investigations showed that gyrB gene sequence data correlated well with DNA-DNA hybridization and phenotypic data, but that 16S rRNA gene sequence data were not suitable for defining species within the genus Photorhabdus. Australian clinical isolates proved to be related most closely to clinical isolates from the USA, but the two groups were distinct. A novel subspecies, Photorhabdus asymbiotica subsp. australis subsp. nov. (type strain, 9802892T=CIP 108025T=ACM 5210T), is proposed, with the concomitant creation of Photorhabdus asymbiotica subsp. asymbiotica subsp. nov. Analysis of gyrB sequences, coupled with previously published data on DNA-DNA hybridization and PCR-RFLP analysis of the 16S rRNA gene, indicated that there are more than the three subspecies of Photorhabdus luminescens that have been described and confirmed the validity of the previously proposed subdivision of Photorhabdus temperata. Although a non-luminescent, symbiotic isolate clustered consistently with P. asymbiotica in gyrB phylogenetic analyses, DNA-DNA hybridization indicated that this isolate does not belong to the species P. asymbiotica and that there is a clear distinction between symbiotic and clinical species of Photorhabdus.

Antimicrobial resistance and genomic screening of clinical isolates of thermophilic Campylobacter spp. from south-east Queensland, Australia.

AIM: To screen 90 clinical isolates of thermophilic Campylobacter species for putative resistance to ampicillin, erythromycin and tetracycline and perform numerical analysis to determine isolate relatedness. METHODS AND RESULTS: Disc diffusion, E-test MIC and agar dilution methods were performed. Disc diffusion testing showed 87 (97%) isolates appeared resistant to ampicillin at 10 microg; 14 (16%) resistant to tetracycline at 30 microg; and three (3.4%) resistant to erythromycin at 15 microg. E-test MICs showed a range of 0.5 to >256 mg l(-1) for ampicillin; 16 to >256 mg l(-1) for tetracycline; and >256 mg l(-1) for erythromycin. E-test showed 68% correlation (+/-1 log2 dilution) with agar dilution for ampicillin, 100% for erythromycin and 64% for tetracycline. Disc diffusion testing showed 100% correlation with agar dilution for erythromycin and tetracycline, and 77% for ampicillin. Numerical analyses of restriction endonuclease (RE) fragment profiles suggested a high level of isolate variation. CONCLUSION: The incidence of resistance of thermophilic Campylobacter spp. to erythromycin and tetracycline is low in south-east Queensland. SIGNIFICANCE AND IMPACT OF THE STUDY: Disc diffusion susceptibility testing may be used to screen thermophilic Campylobacter spp. for putative resistance to erythromycin and tetracycline. Agar dilution should be used to determine ampicillin susceptibility.

Recurrent bacteremia and multifocal lower limb cellulitis due to Helicobacter-like organisms in a patient with X-linked hypogammaglobulinemia.

We describe a 27-year-old man with X-linked (Bruton's) hypogammaglobulinemia who presented during a 10-month period with recurrent fevers and multifocal lower-limb cellulitis associated with bacteremia due to Helicobacter-like organisms ("Flexispira rappini" and Helicobacter canis). Susceptible individuals may acquire infection of this type as a result of exposure to young dogs.

Specific identification, grouping and differentiation of Campylobacter jejuni among thermophilic campylobacters using multiplex PCR.

Campylobacter species such as C. jejuni and C. coli are recognized as major causes of acute gastroenteritis world-wide. Although C. jejuni and C. coli are usually non-pathogenic in birds and animals, they cause enteric disease in humans and the source of infection is often the consumption of contaminated foodstuffs. In this paper we report on the development and use of a multiplex PCR of C. jejuni genomic DNA which yielded a PCR product with a unique polymorphic site that can be used to quickly and accurately identify and group C. Jejuni isolates from any source including DNA, cell culture, skin washings and faecal samples. The test is simple and sensitive and can detect purified DNA from a single bacterium 10(2) cells from crude lysates, 10(3) cells in seeded faeces and 120 cells/ml of washing of 1 cm2 skin fragments of chicken skin.

Isolation, identification, and molecular characterization of strains of Photorhabdus luminescens from infected humans in Australia.

We describe the isolation of Photorhabdus (Xenorhabdus) luminescens from four Australian patients: two with multiple skin lesions, one with bacteremia only, and one with disseminated infection. One of the patients had multiple skin lesions following the bite of a spider, while the lesions in the other patient were possibly associated with a spider bite. The source of infection for the remaining two patients is unknown. As a member of the family Enterobacteriaceae, P. luminescens is unusual in that it fails to reduce nitrate and ferments only glucose and mannose. It gives negative reactions for lysine decarboxylase, arginine dihydrolase, and ornithine decarboxylase (Moeller). The species is motile, utilizes citrate, hydrolyzes urea, and usually produces a unique type of annular hemolysis on sheep blood agar plates incubated at 25 degrees C. A weak bioluminescence is the defining characteristic. P. luminescens is an insect pathogen and is symbiotically associated with entomopathogenic nematodes. Its isolation from human clinical specimens has been reported previously from the United States. Restriction fragment length polymorphism-PCR analysis of the 16S rRNA gene demonstrated a high level of similarity among the Australian clinical strains and significant differences between the Australian clinical strains and the U.S. clinical strains. However, numerical analyses of the data suggest that the two groups of clinical strains are more similar to each other than they are to the symbiotic strains found in nematodes. This is the first report of the isolation of P. luminescens from infected humans in Australia and the second report of the isolation of this species from infected humans worldwide.

Seroepidemiology of invasive pneumococcal disease in Queensland, 1990 to 1997.

Serotypes responsible for 842 cases of invasive pneumococcal disease in Queensland between February 1990 and October 1997 were identified. Type 14 caused 37.5% of episodes in children aged 0-4 years and 19.2% of adult cases. Types 6A, 6B, 14, 18C, and 19F were significantly more frequent in young children while types 3, 4, 7F, 9V and 23F predominated in adults. The regional incidence of type 14 and 7F disease differed significantly in Southeast and Far North Queensland. Coverage for 87% of children aged less than 5 years in this study would be provided by a recently advocated polysaccharide-protein conjugate vaccine containing capsular antigens of types 4, 6B, 9V, 14, 18C, 19F and 23F. Similarly, more than 90% of adults would be covered by the currently available 23- valent polysaccaride vaccine.