First Siena International Conference on Animal Models of Chronic Obstructive Pulmonary Disease, Certosa di Pontignano, University of Siena, Italy, September 30-October 2, 2001

The meeting proved of great interest to those developing an animal model of chronic obstructive pulmonary disease (COPD). COPD is caused by cigarette smoking, evidenced by deterioration in lung function. Lung function is only rarely assessed in animal models. A cigarette smoke driven pathology should provide the best in vivo model for COPD. However, as lesions produced this way take 8–12 months to develop other strategies have to be employed. Emphysematous lesions were also achieved by treatment with elastase, lipopolysaccharide, ozone and other inducers. Several studies described treatments that have shown activity in these models. Transgenic models were discussed, as was the importance of species and strain selection.     

Cancer molecular epidemiology: new horizons of prophylaxis

Perspectives of malignant neoplasm prophylaxis based on molecular biology achievements are discussed. Gene variants critical to development of hereditary cancer syndromes, genes modulating malignant neoplasm development risk without hereditary cancer syndrome development, and genes determining tendency of individuals for different malignant neoplasm progress risk increasing lifestyle factors are examined. Molecular epidemiology by using large scale population analysis of cancerogenesis linked genetic polymorphisms prevalence allows determination of risk groups at the most earlier stages of cell transformation or even before the onset of cell malignization and development of goal-based prophylaxis measures based on polymorphism and corresponding cancer type. Epidemiologic analysis of this type allows for earlier diagnostics in risk groups, therapy efficacy increase, disability decrease. Specific therapy on molecular level may be possible in the future.     

Identification of Malassezia species isolated from patients with extensive forms of pityriasis versicolor in Siena,Italy

BackgroundPityriasis versicolor (PV) is an infection caused by various species of Malassezia yeast. There is no agreement in the literature concerning the species of Malassezia and the demographic, clinical, and mycological data.AimsTo prospectively identify Malassezia species isolated from lesions of patients with extensive, long standing and recurrent forms of PV and to estimate the relationship between Malassezia species and the demographic and clinical data of the patients.MethodsAll patients with PV were enrolled over a four-year period. Malassezia species were isolated in cultures and identified by morphological features and physiological tests. In the last 2 years a PCR-based technique was used to confirm the species’ identification.ResultsA total of 74 patients (43 males and 31 females, mean age 39.5 years) were enrolled. Only one species was isolated in 45 patients, and more than one species were identified in the remaining 28 patients (38%). M. globosa was the most frequently isolated (60.3%) species. There was a significant association between the isolation of 2 or more species and the presence of at least one predisposing factor. In the last 29 cases, which were subjected to PCR, there were no differences in the identification of isolated species as compared to traditional methods.ConclusionsThe isolation of more than one species in a single lesion is not infrequent in PV and is related to the presence of one predisposing factor. The isolated species isolated were not influenced by demographic and clinical features. The traditional and more recent (PCR) procedures gave the same results in the isolated species.     

Rubella in Russia: variability of the infectious agent during the period of vaccine prophylaxis

AIM: To assess influence of vaccination against rubella on the genetic diversity of rubella virus. MATERIALS AND METHODS: Vaccine strains of rubella virus Wistar 27/3 and Orlov-B as well as sera from patients with rubella obtained in Perm region during 1999 - 2005 period and standard serologic, molecular, epidemiologic and statistical methods were used. The study was performed according to the WHO recommendations on the genotyping of wild rubellavirus strains. RESULTS: Strains of rubella virus isolated in Perm region, vaccine strain Orlov-B (Saint Petersburg), and 4 Russian strains isolated in 1967-1997 before vaccine introduction belong to the same genetic group with high degree of homology - genetic divergence do not exceed 0 - 1%. This group was identified as genotype 2c which, according to WHO's data, circulates only in Russia. Periods of epidemic peaks of rubella incidence and its falls as well as selective immunization of girls and women of childbearing age did not influence on the genetic stability of the virus (divergence did not exceed 0.6 - 2.0%). On the contrary, mass immunization of children aged 1 - 2 years during 4 years resulted in statistically significant changes of rubella virus subtype inside the genotype 2c. CONCLUSION: Ten-year experience of rubella vaccination in Perm region demonstrates necessity of inclusion of monitoring for rubella virus variability in the system of epidemiological surveillance and control for rubella infection during period of its prevention by vaccine.     

Coenological research on macrofungi in evergreen oak woods in the hills near Siena (Italy)

For a mycocoenological study of macromycetes in Mediterranean evergreen forests in the Siencese province, five stands belonging to theQuercion ilicis have been observed during more than two years. The phytosociological analysis of these evergreen oak woods, situated at the limit of the geographical distribution of the alliance, reveals them as intermediate between the typical Mediterranean vegetation and the sub-Mediterranean deciduous broad-leaved forests. 181 fungal species were recognized and assigned to seven ecological groups on the basis of substratum, habitat, forest management and/or particular mycorrhizal relationship. Our results have been compared with those obtained in some European deciduous forests. Such a comparison shows the occurrence of 16 thermophilous fungal species in the Siena woods which are missing or very rare in the European deciduous forests. On the basis of our present knowledge, five of these species (Boletus lepidus, Hygrophorus dichrous, Hygrocybe nigrescens, Inocybe similis, Phellinus torulosus) may be considered as strictly tied to the evergreen oak woods. Five others, which have been reported for several European phytocoenoses, seem to be ‘preferential taxa’ of our evergreen oak woods, namely:cortinarius calochrous, C. sodagnitus, Hygrophorus arbustivus, H. russula, Lyophyllum immundum. On the whole the highest similarities have been found with the more xerothermophilous European forests (Querco-Lithospermetum andSorbo-Quercetum). The presence in theQuercus ilex woods of numerous more mesophilous fungi, some of which are widespread in beech forests, underlines the transitional nature of the phytocoenoses studied.     

Modern features of the epidemiology and prophylaxis of influenza

Modern concepts concerning influenza pandemics and epidemics in different countries of the world are presented. The influenza epidemics of the last decade in different countries of the world and their specific features linked with the "drifting" variability of influenza virus have been analyzed. Information on influenza morbidity during the last 30 years is given; on the basis of this information the role of vaccinal prophylaxis and mainly the mass vaccination of school children and students, is shown. The results of the efficacy of such vaccines as live influenza vaccine, American split vaccine, Russian live recombinant vaccine and Grippovac (1995-1996), as well as new-generation vaccine Grippol (1998), are presented. The prospects of the combined use of specific and unspecific prophylaxis have been determined.     

Tick-borne diseases in ruminants of Central and Southern Italy: epidemiology and case reports

Sera and blood from cattle and sheep were examined for the presence of Babesia and Theileria spp by microscopy and serology at the Parasitology Department of the Istituto Zooprofilattico Sperimentale of Abruzzo and Molise (IZSAM). Of the 47 bovine herds (323 animals) tested, 15 were found positive for Babesia bigemina and 1 for Babesia bovis. Two outbreaks occurred, one caused by B. bigemina and one by B. bovis. The B. bigemina outbreak occurred in Abruzzo and has been followed for two years. The isolate of B. bigemina was very pathogenic leading to the death of two cows out of 57. The vector responsible of the transmission appeared to be Rhipicephalus bursa. Parasites were observed in the erythrocytes for 30 days whereas sera were positive to indirect immunofluorescence (IFA) for at least one year. The B. bovis outbreak occurred in the province of Mantova (Northern Italy) in a group of 70 beef cattle imported from France. The infection resulted in the death of 5 animals and severe illness in another 6. In contrast with what occurred for Babesia infection, no clinical cases were recorded in cattle when species of Theileria were detected by microscopy. Of the 24 bovine herds (252 animals) tested for Theileria, 21 were found positive for the T. "sergenti"/buffeli/orientalis group. Single and mixed infection of T. "sergenti" and T. buffeli/orientalis were detected in herds of cross-bred cattle from Abruzzo and Marche. The parasites were identified by using a polymerase chain reaction which amplified DNA encoding p32/34. Most of the collected ticks (90%) were adults of R. bursa whereas the others were adults of Hyalomma detritum. During the period the animals have been observed (18 months), no clinical cases have been recorded and no associations have been found between blood abnormalities and animals found infected with Theileria. Prevalences of subclinically infected carriers increased from February till December (95.4%) even if the animals were indoors and no ticks were present. The prevalence then dropped dramatically six months later (76.7%). In calves less than 1 year old, the prevalence of infection significantly (p<0.05) increased with age, however intraerythrocytic stages of Theileria were found in the blood of three newborn calves (<7 days of age). Of the 18 ovine flocks tested for Babesia spp. (150 animals examined), 1 was positive for B. ovis and 2 for B. motasi. B. motasi infection was not associated with symptoms, while an outbreak of babesiosis caused by B. ovis occurred in Abruzzo. The infection resulted in the death of 3 animals (0.75% of the flock), two rams (20% of the total number) and a ewe, and severe illness in another 5 ewes (2% of the flock). Specimens of R. bursa and R. turanicus were collected from the infected animals. Of the 18 flocks (150 animals) examined, 12 were microscopically positive for Theileria spp. No clinical cases were recorded and identification at species level was not possible on the basis of morphological criteria. The prevalence distribution of infected herds and infected animals within herds and flocks have been calculated by a Monte Carlo simulation model, running 10,000 iterations. The most likely levels of prevalence of infected herds and infected animals within herds found for the species observed were as follows: 20% for B. bigemina with a prevalence within herd of 27%, 11% for B. bovis (18% within herd), 10% for Babesia ovis (19% within herd), 10% for B. motasi (17.5% within herd), 63% for Theileria in cattle (66% within herd) and 51% for Theileria in sheep (55% within herd).     

Descriptive epidemiology of a scabies epidemic in chamois in the Dolomite Alps,Italy

In 1995, the hitherto Sarcoptes-free alpine chamois (Rupicapra rupicapra) of the Dolomite Alps (Italy) were affected by scabies for the first time after the spread of the disease from a neighbouring focus in Austria. Since then, four Agencies cooperated to warrant monitoring of the spatial and temporal progress of the outbreak and further active surveillance was carried out within an intensive study area. In ten years, 15 meta-populations numbering 10,000 chamois and 210,000 ha were encompassed with a maximum distance of 55 km from the index case. “Oil spot” advancement of the epidemic was observed together with “jumps” of 9 to 20 km followed by spreading of the infection to the intervening population units. Demographic decline of the four meta-populations so far affected for a minimum of six years ranged between 49 and 77% (), whereas similar post-epidemic densities of 1.1 to 1.7 heads/100 ha () were recorded. Contiguous resistant and sensitive herds with similar pre-epidemic density were detected on a smaller population scale, suggesting complementary mechanisms intervening besides density-dependence as determinants of the outbreak outcome. No sex or age class showed a higher sensitivity to scabies out of a sample of 1,696 infected chamois. A peak prevalence of free-ranging chamois bearing overt scabietic lesions was observed in January and February. Cases were only sporadically diagnosed in other sympatrc wild ruminants (Cervus elaphus, Capreolus capreolus and Ovis gmelini musimon), whereas all four alpine ibex (Capra ibex ibex) herds living in the surveyed area suffered from scabies-induced decline.     

Deterioration of medieval painting in the chapel of the Holy Nail, Siena (Italy) partially treated with Paraloid B72

Painted surfaces in the Chapel of the Holy Nail in the medieval ex-hospital of Siena show five areas of oxidation and blackening. Most were treated with Paraloid B72 acrylic resin 40 years ago. To study deterioration of the painted surfaces by scanning microscopy with X-ray dispersion microanalysis, fragments obtained from the five areas were analyzed directly and after hydration. Hydrated fragments treated with Paraloid showed a compact, shiny, impermeable surface. The acrylic resin forms a layer that inhibits interaction of the painting with the environment, including transpiration and gaseous exchange. Element composition of paint layers of different colours revealed gold, iron (oxides in ochre) and lead (oxide) tempered with lime and/or gypsum. A few microbial cells were found on all fragments. To study biological deterioration in the five areas, fragments were hydrated and incubated in minimal culture medium without added carbon source to select bacteria capable of using carbon sources in the medieval fresco. Analysis did not reveal any spores or hyphae, and excluded physical and mechanical damage. Bacteria of the genus Bacillus were isolated only from untreated samples. Paraloid resin forms an inert film over the mural, preventing access to microbes able to use nutrient sources in the painted layer. The screening of 16S rRNA libraries from enrichment cultures showed wide phylogenetic diversity. Forty-four percent of the clones retrieved from the clone library were affiliated with the order Firmicutes, confirming the prevalence of aerobic spore-formers among the colonizing microflora. Firmicutes therefore presumably produced extracellular material which made water available to other bacteria which may have converted thiosulphate in the medieval tempera to sulphur globules. Sulphur is a known oxidant of metallic pigments, in this case aluminium, which may explain the blackening of the untreated sample.     

Rubella: immunity and vaccination in schoolgirls.

Of 191 schoolgirls, 128 volunteered to take part in a feasibility study of serotesting before and after rubella vaccination, and all responded to RA 27/3 vaccine. Had the serum samples been taken by a fingerprick method the number of volunteers would probably have increased considerably. A change in policy for rubella vaccination to testing both before and after vaccination would cost no more than the existing policy, would ensure primary response, and would differentiate those women who were protected by the vaccine from those with antibody to wild virus.