Cats are the natural host for Bartonella henselae, an opportunistic human pathogen and the agent of cat scratch disease. Here, we have analyzed the natural variation in gene content and genome structure of 38 Bartonella henselae strains isolated from cats and humans by comparative genome hybridizations to microarrays and probe hybridizations to pulsed-field gel electrophoresis (PFGE) blots. The variation in gene content was modest and confined to the prophage and the genomic islands, whereas the PFGE analyses indicated extensive rearrangements across the terminus of replication with breakpoints in areas of the genomic islands. We observed no difference in gene content or structure between feline and human strains. Rather, the results suggest multiple sources of human infection from feline B. henselae strains of diverse genotypes. Additionally, the microarray hybridizations revealed DNA amplification in some strains in the so-called chromosome II-like region. The amplified segments were centered at a position corresponding to a putative phage replication initiation site and increased in size with the duration of cultivation. We hypothesize that the variable gene pool in the B. henselae population plays an important role in the establishment of long-term persistent infection in the natural host by promoting antigenic variation and escape from the host immune response.

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Bartonella henselae is present in a wide range of wild and domestic feline hosts and causes cat-scratch disease and bacillary angiomatosis in humans. We have estimated here the gene content of Bartonella koehlerae, a novel species isolated from cats that was recently identified as an agent of human endocarditis. The investigation was accomplished by comparative genomic hybridization (CGH) to a microarray constructed from the sequenced 1.93-Mb genome of B. henselae. Control hybridizations of labeled DNA from the human pathogen Bartonella quintana with a reduced genome of 1.58 Mb were performed to evaluate the accuracy of the array for genes with known levels of sequence divergence. Genome size estimates of B. koehlerae by pulsed-field gel electrophoresis matched that calculated by the CGH, indicating a genome of 1.7 to 1.8 Mb with few unique genes. As in B. quintana, sequences in the prophage and the genomic islands were reported absent in B. koehlerae. In addition, sequence variability was recorded in the chromosome II-like region, where B. koehlerae showed an intermediate retention pattern of both coding and noncoding sequences. Although most of the genes missing in B. koehlerae are also absent from B. quintana, its phylogenetic placement near B. henselae suggests independent deletion events, indicating that host specificity is not solely attributed to genes in the genomic islands. Rather, the results underscore the instability of the genomic islands even within bacterial populations adapted to the same host-vector system, as in the case of B. henselae and B. koehlerae.

A sterile white fungus was isolated from the healthy looking roots of buffalo grass (Stenotaphrum secundatum) grown on cleared bush land in Perth, Western Australia. The fungal strain was pathogenic on 12 plant species screened under the greenhouse conditions. The clamp connections and dolipore septa indicated that the isolate was a Basidiomycete. Mycelial features, growth rate at different temperatures, as well as pathogenicity patterns of this sterile white basidiomycete (SWB) were distinctly different from those of a strain with a similar morphology, ATCC 28344, previously described as a pathogen in Florida and Georgia (USA). All attempts to induce sporulation failed. The isolates were also compared using the nucleotide sequence analysis of the ribosomal DNA array. Approximately 1 kbp of the 5¢ end of the large subunit ribosomal RNA gene, complete sequences of the small subunit ribosomal RNA gene and the entire ITS region (including ITS1, ITS2 and 5.8S gene) were sequenced for the purpose. The obtained sequences were compared with the homologous regions of other genera of Agaricales available in GenBank. Relatively low sequence similarities between the American and Australian strains, as well as the phylogenetic analysis of the studied regions has suggested that these two fungi belong to different genera. Interesting results were achieved in the case of the large subunit ribosomal DNA since this region has been widely studied for taxonomy of Basidiomycetes. The Australian strain 3034 appeared to be closely related to the genus Campanella and the American SWB was identified as belonging to the genus Marasmius, possibly to M. graminum. Our data suggest that the Australian strain is a novel pathogen, and is different from the American SWB isolates described to date.Abbreviations: SWB -sterile white basidiomycete

Rhododendron cultivation has a long history in northern Italy where a wide selection of varieties and hybrids are grown. In the fall of 2002, a previously unknown disease was observed on Azalea japonica cv. Palestrina in several commercial farms in the Province of Verbania. Leaves showed irregular necrotic areas that were 1 mm in diameter. Lesions were dark brown to black and were surrounded by a chlorotic halo. Eventually, lesions coalesced, forming large irregular spots. Heavily infected leaves fell prematurely, resulting in severe defoliation. On the infected leaves, acervuli were present from which cylindrical tapered conidia measuring 4.8 to 7.2 × 11.0 to 22.8 μm at one end were released. Fifty conidia per isolate were measured. Fungus identified as Colletotrichum acutatum was consistently recovered from infected leaves, disinfested in 1% NaOCl for 1 min, and plated on potato dextrose agar amended with 100 mg/l of streptomycin sulfate. Pathogenicity of three fungal isolates was confirmed by inoculating healthy A. japonica (cvs. Palestrina and Snow) plants grown in plastic pots (18-cm diameter, 3 liters). Plants (five per treatment) were sprayed with a conidial suspension (1 × 106 conidia per ml) of the three isolates of C. acutatum. Noninoculated plants served as a control. Inoculated and control plants were covered with plastic bags to maintain high relative humidity conditions. All plants were maintained in growth chambers at 20 ± 1°C (12 h per day of fluorescent light). Six days after the artificial inoculation, plants developed typical symptoms on the leaves. C. acutatum was consistently reisolated from infected plants. The pathogenicity test was carried out twice. Sequencing of the internal transcribed spacer region of the rDNA and a portion of the β-tubulin gene were performed, and the obtained sequences were compared with those available in GenBank. Identification of the fungus as C. acutatum, therefore was confirmed. To our knowledge, this is the first report of the presence of C. acutatum on A. japonica in Italy. Although observed presently only in a few nurseries, the disease has the potential to spread, becoming more relevant in an area where rhododendron cultivation is economically important. Anthracnose on rhododendron has already been described in several countries (1,2). References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (2) O. Vinnere et al. Mycol. Res. 106:60, 2002.