No abstract

This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues.Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. a b s t r a c tBlackened areas on outdoor marble statues are a significant esthetic problem due to the presence of deteriorating agents. Microcolonial black fungi, which have their natural ecological niche on rocks, play an important role in deterioration of stones used in monuments, such as marble and limestone. Black fungi were isolated from two very valuable statues exposed to the outdoor environment in Florence, Italy, the "Ratto delle Sabine" and the "Copia del David," and these fungi were demonstrated to be responsible for the blackening areas on the statues. The black strains showed many features common to members of rock-inhabitants dematiaceous fungi. Morphological and molecular characterization, including phylogenetic analysis, indicated that the strains isolated from both statues and in different times belong to the same species and can be assigned to the rock-inhabitant genera Sarcinomyces and Phaeococcomyces. Red yeasts growing in close proximity to the black ones, with no visible effect on the statues, were also characterized on the morphological and molecular level and identified as Sporobolomyces yunnanensis.

The strain diversity and the population structure of nosocomial Acinetobacter isolated from patients admitted to different hospitals in Florence, Italy, during a 3-year surveillance program, were investigated by amplified fragment length polymorphism (AFLP). The majority of isolates (84.5%) were identified as A. baumannii, confirming this species as the most common hospital Acinetobacter. Three very distinct A. baumannii clonal groups (A1, A2, and A3) were defined. The A1 isolates appeared to be genetically related to the wellcharacterized European EU II clone. A2 was responsible for three outbreaks which occurred in two intensive care units. Space/time population dynamic analysis showed that A1 and A2 were successful nosocomial clones. Most of the A. baumannnii isolates were imipenem resistant. The genetic determinants of carbapenem resistance were investigated by multiplex PCR, showing that resistance, independently of hospital origin, period of isolation, or clonal group, was associated with the presence of a bla OXA-58-like gene and with ISAba2 and ISAba3 elements flanking this gene. bla OXA-58 appeared to be horizontally transferred. This study showed that the high discriminatory power of AFLP is useful for identification and typing of nosocomial Acinetobacter isolates. Moreover the use of AFLP in a real-time surveillance program allowed us the recognition of clinically relevant and widespread clones and their monitoring in hospital settings. The correlation between clone diffusion, imipenem resistance, and the presence of the bla OXA-58-like gene is discussed.

Seiridium cardinale is regarded as the most important agent responsible for the disease of cupressaceous hosts referred to as Cypress canker. The fungus was first described in California and is currently reported in all continents. A recent study based on seven SSR loci has suggested that California populations may represent the source of the epidemic in the Mediterranean. In this study, 185 AFLP markers were used on an expanded sample size of 125 isolates to determine whether the Mediterranean population may indeed be derived from the California one and to compare the reproductive modes of populations in the two regions of the world. Additionally, AMOVA, NJ, STRUC-TURE, principal component analysis and b-tubulin sequence analyses were employed to infer the presence of genetic structure within and between populations. The distribution of pairwise AFLP similarity coefficients suggests Mediterranean populations are reproducing only clonally, while California populations are reproducing both clonally and sexually. AMOVA indicates Mediterranean and California populations are currently genetically isolated, but NJ and STRUCTURE analyses both suggest ancestral Mediterranean genotypes belong to the California population. No alleles were private to either population, and the presence of identical or quasi-identical genotypes at large distances supports the notion that movement of infected cypress plants is responsible for the global spread of the disease. Surprisingly, STRUCTURE identified a second cluster of Mediterranean genotypes distinct from the basal mixed California-Mediterranean cluster. This second cluster either may have originated from the first one under the selection pressure imposed on the pathogen in the new Mediterranean environments or may be the result of a further introduction from California or elsewhere. Cumulatively, the evidence presented here suggests that S. cardinale may be native or long naturalized in California and that two genetically distinct groups are present in the Mediterranean, with obvious implications for further studies on this disease.

We report the results of a three-year surveillance program of Klebsiella spp. in six hospitals in Florence (Italy). A total of 172 Klebsiella isolates were identified and typed by AFLP: 122 were K. pneumoniae and 50 were K. oxytoca. Most K. pneumoniae (80%) and K. oxytoca (93%) showed unrelated AFLP profiles. Beside this heterogeneous population structure, we found five small epidemic clonal groups of K. pneumoniae. Four of these groups were involved in outbreak events, three of which occurred in neonatal ICUs. The fifth clonal group spread in three different wards of two hospitals. Only one non-epidemic clonal group of K. oxytoca was detected. The frequencies of isolates with multiple antibiotic resistances increased with time; at the end of the study period, most K. pneumoniae were resistant to all the antibiotics tested. A PCR analysis of seven ertapenem resistant isolates was unable to detect any of the major genes known to underlie carbapenem resistance in K. pneumoniae.