First Isolation of Pseudogymnoascus destructans, the Fungal Causative Agent of White-Nose Disease, in Bats from Italy.

White-nose disease, caused by the dermatophyte Pseudogymnoascus destructans, is a devastating pathology that has caused a massive decline in the US bat populations. In Europe, this fungus and the related infection in bats have been recorded in several countries and for many bat species, although no mass mortality has been detected. This study reports for the first time the presence of P. destructans in Italy. The fungus was isolated in the Rio Martino cave, a site located in the Western Alps and included in the Natura 2000 network. Twenty bats, belonging to five different species, were analysed. The fungus was retrieved on eight individuals of Myotis emarginatus. The allied keratolytic species P. pannorum was observed on two other individuals, also belonging to M. emarginatus. Strains were isolated in pure culture and characterized morphologically. Results were validated through molecular analyses. Future work should be dedicated to understand the distribution and the effects of the two Pseudogymnoascus species on Italian bats.

Chemical composition and antimicrobial activity of Phyllanthus muellerianus (Kuntze) Excel essential oil.

ETHNOPHARMACOLOGICAL RELEVANCE: Phyllanthus muellerianus (Kuntze) Excel (family Euphorbiaceae) stem bark is used in Cameroon by Baka pygmies as a remedy for wound healing and tetanus. AIM OF THE STUDY: To characterize the chemical composition and to evaluate the antimicrobial properties of the essential oil of the plant. MATERIALS AND METHODS: The essential oil was extracted from the stem bark by dynamic head space and by hydrodistillation and characterized by GC and GC-MS analyses. The antimicrobial activity was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal-fungicidal concentration (MBC-MFC) by the micro and macrodilution methods. The following bacteria and fungi were used: Clostridium sporogenes ATCC 3584, Staphylococcus aureus ATCC 6538, Streptococcus mutans ATCC 25175, Streptococcus pyogenes ATCC 19615, Escherichia coli ATCC 10536, Candida albicans ATCC 10231, Candida albicans LM 450, Trichophyton mentagrophytes LM 230, Trichophyton rubrum LM 237, Microsporum canis LM 324. RESULTS: The hydrodistillation afforded 0.06% (dry weight basis) of pale yellow oil. Thirty-eight compounds representing 90.69% were identified. The major component (36.40%) was found to be (E)-isoelemicin, identified by comparison of its (1)H-NMR experimental data, with literature data. The oil showed good antibacterial activity against Clostridium Sporogenes, Streptococcus mutans and Streptococcus pyogenes with MIC ranging from 13.5 to 126 µg/ml. A weak antifungal activity (MIC 250 µg/ml) was found against Trichophyton rubrum, only. CONCLUSIONS: The antimicrobial activity and the chemical composition of Phyllanthus muellerianus stem bark essential oil are reported for the first time.

Pyrogenic activity of air to characterize bioaerosol exposure in public buildings: a pilot study.

AIMS: The aim of our study was to investigate indoor air quality (IAQ) by comparing pyrogen concentration and microbiological contamination in offices in public buildings. METHODS AND RESULTS: Air samples were collected during cold and warm seasons in 39 offices in four European cities. Pyrogens were measured by the in vitro pyrogen test (IPT), moulds and bacteria by classical microbiology. In 92% of the investigated offices, pyrogen and microbial contaminations were below 150 EEU m(-3) and 10(3) CFU m(-3), respectively, whilst in 75%, moulds did not exceed 10(2) CFU m(-3). CONCLUSIONS: The IPT is a rapid, reliable tool for measuring pyrogens that could be used as an 'early warning' indicator of IAQ. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on pyrogenic compound detection in offices using IPT, which could serve for developing future indoor air guidelines.

Determination of aerial microbiological contamination in scholastic sports environments.

AIMS: To assess the microbiological indoor air quality (IAQ) of high school and college gyms during physical training lessons and to evaluate the effective microbiological exposure of students. METHODS AND RESULTS: Air samples from 11 high school and college gyms of Pavia, Italy were collected. Total bacterial counts, total fungal counts and characterization of fungal taxa were determined. Air quality was evaluated using three microbiological contamination indices: the global index of microbiological contamination per m3 (GIMC per m3), the index of mesophilic bacterial contamination (IMC) and the amplification index (AI). CONCLUSIONS: This work testifies that air contamination in indoor gyms is always superior to that of the outdoor environment. Nevertheless, students are exposed to relatively low concentrations of airborne micro-organisms. The highest values of fungal counts and GIMC per m3 (>14,661) were observed between April and October when the central heating systems were switched off. The lowest fungal counts were detected in modern buildings equipped with forced ventilation systems. From qualitative aeromycological studies, 45 fungal taxa were identified, and different potentially allergenic species were isolated. SIGNIFICANCE AND IMPACT OF THE STUDY: The standardization of air sampling methods and the correct evaluation of aeromicrobiological results allow the classification of indoor air healthiness. The proposed microbiological contamination indices together with the characterization of airborne fungal taxa are useful tools for detailed description of IAQ.

First Report of Gray Leaf Spot on St. Augustinegrass in Italy.

St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) is used for lawns in southern Italy because it is much more resistant to biotic and abiotic adversities than other turfgrass species. Because few seeds are viable, this species is established by vegetative propagation. A new disease was noticed during the spring of 2002 and 2003 on cuttings of St. Augustinegrass growing in three greenhouses in eastern Sicily. The disease affected leaves and culms and caused a progressive drying of the plants. The infection was first seen on leaves as gray, necrotic spots that enlarged in high-humidity conditions to form oval, and later, spindle-shaped lesions. In association with the lesions, it was possible to observe fungal spore development and sunken areas with blue-gray centers and slightly irregular, brown margins with yellow halos. Spots were concentrated without specific arrangement along longitudinal veins and the midrib and at the base, tip, and margins of the leaf blade. Symptoms on the culms consisted of brown-to-black blotches that sometimes extended throughout the internodes. From these infected tissues, 20 explants taken from leaves and culms were cut, washed with sterile water, and placed on 1.5% water agar (WA). Later, conidia and conidiophores were obtained from colonies with a sterile glass needle and placed on 4% WA. From these plates, two monoconidial isolates were obtained and transferred to rice meal medium (1). The colonies were identified as Pyricularia grisea Cooke (Sacc.), anamorphic state of Magnaporthe grisea (Hebert) Yeagashi & Udagawa, the cause of rice blast disease and gray leaf spot disease of turfgrasses. The conidia were pyriform to obclavate, narrowed toward the tip, rounded at the base, 2-septate, 21 to 31 µm × 6 to 10 µm (average 25.7 ×8.2 µm). Pathogenicity tests were performed by inoculating leaves and culms of six St. Augustinegrass plants with a conidial suspension of the fungus (1.5 ×105 conidia per ml). The same number of noninoculated plants was used as controls. All plants were incubated in a moist chamber with high humidity at 25°C. After 6 days, all inoculated plants showed typical symptoms of the disease. Koch's postulates were fulfilled by isolating P. grisea from inoculated plants. Gray leaf spot caused by P. grisea has been a chronic problem on St. Augustinegrass since it was first reported in 1957 (2). To our knowledge, this is the first report of P. grisea on St. Augustinegrass in Italy. While it does not appear to be an important disease in the field at this time in Sicily, it could cause losses in greenhouses where vegetative material is propagated for field planting. A preliminary molecular analysis has shown a clear distinction between the tested strain and other strains isolated from rice seeds and plants in northern Italy. References: (1) E. Roumen et al. Eur. J. Plant Pathol. 103:363, 1997. (2) L. P. Tredway et al. Plant Dis. 87:435, 2003.

Superficial mycoses in Italy.

A total of 1244 cases suspected of superficial mycoses were analysed in a period of 12 months in Italy. Dermatophytoses were found in 62.4%; the other common disease was Pityriasis versicolor (44.1%)., The predominant species Epidermophyton floccosum (34,2%) was strikingly high in relation to other statistics; this species was commonest in tinea cruris. Among the other dermatophytes M. canis was predominant (31.3%) commonest in tinea corporis, followed by T. mentagrophytes (17,8%) and T. rubrum (10,3%). A high rate of yeasts was found in tinea manum (81.6%) and tinea pedis (47.7%). ?

Multicentre clinical study with tolciclate in the local treatment of skin mycoses in 1083 patients.

A multicentre clinical trial with tolciclate was carried out in Italy on 1083 patients suffering from skin mycoses (tinea corporis, tinea cruris, tinea pedis, tinea manuum and pityriasis versicolor). Both preparations (1% cream and lotion) showed a good activity evaluated weekly by clinical examinations and mycological assessments, i.e. culture for tineas and microscopy for pityriasis versicolor. Favourable clinical results ranged from 83% to 97% according to diagnoses. Cultural or microscopic conversions obtained in a mean time of about 2 weeks varied from 70% to 91%. Mycological relapses a month after the end of treatment were seen in 6.5% of cases examined. Adverse reactions were observed in 5.6% of patients but the treatment was discontinued only in 2.6%.