First Report of Sydowia polyspora on Aleppo Pine (Pinus halepensis) in Italy.

Pinus halepensis Mill. is a pine native to the Mediterranean region that is generally located from sea level up to an altitude of 200 meters. In July 2012, extensive leaf yellowing and scorching were observed on the foliage of two specimens of P. halepensis in a public park of Alassio municipality, Liguria region (northern Italy). Diseased needles showed chlorotic and reddish brown colored areas that were randomly distributed among healthy needles. In order to isolate the potential pathogen, diseased needle tissues were surface sterilized for 1 min in 1% NaOCl and plated onto potato dextrose agar (PDA) amended with 100 ppm of streptomycin sulfate. After 7 to 10 days, a brown to dark green mycelium slowly developed on the PDA. From the mycelium, single cell conidia averaging 7.0 (4.7 to 12.2) × 2.9 (2.4 to 5.4) µm (n = 50) were produced. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS1f/ITS4 and sequenced. A BLAST (1) search yielded 100% of maximum identity with Sydowia polyspora (Bref. & Tav.) E. Müller for ITS1f and ITS4. Pathogenicity was confirmed by inoculating 15 3-year-old plants (approximately 5 months after bud break) of P. halepensis grown singly in 8-cm diameter pots and maintained in greenhouse. Twelve of 15 plants were wounded by gently rubbing needles together. Five non-inoculated plants, with wounded and non-wounded branches, were kept as controls. Inoculations were carried out at 16.5 to 18.5°C. A suspension of S. polyspora conidia was made by adding 1 to 2 ml sterile water to the spore mass on 1-month-old PDA cultures in 9-cm petri dishes. Inoculum (107 spores/ml) was applied on the needles with a soft paint brush. After inoculation plants were covered with polythene bags for 5 days, kept at temperatures ranging from 5.5 to 26.4°C (average 14.5°C), and watered as needed. The inoculation trial was repeated once. All inoculated plants, both wounded and non-wounded, developed leaf yellowing and browning since the fifth day after inoculation in both inoculation tests, while control plants remained symptomless. After 15 days from first symptom appearance, S. polyspora was re-isolated from the needles of inoculated plants according to the procedure already described. S. polyspora is associated with current season needle necrosis (CSNN) on various conifer species. P. halepensis was reported in Spain to be a susceptible host (2) and S. polyspora has been isolated from infected tissues of fir (Abies spp.) (3). To our knowledge, this is the first report of S. polyspora on P. halepensis in Italy. Control of the pathogen with fungicides was ineffective on fir species, while application of very high rates of calcium chloride during shoot elongation was able to reduce the severity of CSNN (3). In forest areas, municipality gardens, and parks, effective management strategies have not yet been developed. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) L. Botella et al. Fungal Diversity 40:1, 2010. (3) V. Talgø et al. Fungal Biol. 114:545, 2010.

First Report of Botrytis Blight Caused by Botrytis cinerea on Chamelaucium uncinatum in Italy.

Chamelaucium uncinatum (wax flower), an evergreen shrub belonging to the Myrtaceae family, is suitable for growing in containers. In the Albenga area (northern Italy), this species is grown as a potted plant. In April 2009, symptoms of a previously unknown blight were observed in a commercial glasshouse in the Savona Province (northern Italy) on 80% of 500 potted plants of cv. Snow Flake. Glasshouse temperatures ranged between 16 and 22°C and plants were drip irrigated. Initially, leaves and calyces appeared chlorotic. Subsequently, necrotic lesions developed on flower stalks and occasionally the corollas. After 10 days, soft, gray mycelium became apparent on symptomatic tissue, especially on the foliage. Severely infected leaves and flowers eventually became completely necrotic and abscised. Tissues were excised from diseased leaves, immersed in a solution containing 1% sodium hypochlorite for 10 s, and then cultured on potato dextrose agar (PDA) medium. A fungus developed abundant mycelium when incubated under constant fluorescent light at 23 ± 1°C. Numerous, small sclerotia also developed on PDA plates incubated for 20 days at 8 ± 1°C. Sclerotia were dark, spheroid, and measured 0.5 to 1.8 × 0.5 to 1.5 (average 1.2 × 1.0) mm. Conidia were smooth, gray, unicellular, ovoid, measured 8.5 to 11.1 × 7.1 to 8.6 (average 9.7 × 7.8) µm, and similar to those described for Botrytis cinerea (2). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 495-bp segment showed 100% similarity with the sequence of Botryotinia fuckeliana (perfect stage of B. cinerea). The nucleotide sequence has been assigned the GenBank Accession No. GQ149477. Pathogenicity tests were performed by spraying leaves of healthy potted C. uncinatum with a spore suspension (2 × 104 conidia/ml) obtained from PDA cultures of the pathogen. Each plant received 30 ml of the inoculum. Plants sprayed with water only served as controls. Three plants per treatment were used. Plants were covered with plastic bags for 5 days after inoculation and maintained in a growth chamber at 20 ± 1°C. The first foliar lesions developed on leaves 7 days after inoculation and were similar to those observed in the commercial glasshouse, whereas control plants remained healthy. B. cinerea was consistently reisolated from these lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of B. cinerea on C. uncinatum in Italy as well as in Europe. The disease has been reported in California (3) and more recently in South Africa (4). In Italy, the economic importance of the disease is currently still limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, England, 1971. (3) A. M. French. California Plant Disease Host Index. Calif. Dep. Food Agric., Sacramento, 1989. (4) L. Swart and S. Coertze. Plant Dis. 86:440, 2002.

Use of IgG Avidity test in case definitions of toxoplasmosis in pregnancy.

A survey network for congenital toxoplasmosis (TOXO-NET) was set up in December 1996 in Piedmont (Italy). Participants were asked to classify the infections in pregnant mothers and newborns by the criteria of the European Network on Congenital Toxoplasmosis published by Lebech in 1996. Because the IgG Avidity test is largely employed as a 2nd level test in toxoplasmosis diagnosis and it could be helpful to date infection, the co-ordinators of TOXO-NET suggested including it in the "case definition" of "probable" infection and "unlikely" infection. 117 cases of toxoplasmosis in pregnancy divided into the risk categories under Lebech's criteria were re-examined using the "new" case definitions. 77 out of 117 (65.8%) Toxoplasma gondii infections during pregnancy could be defined with only one serum sample using the IgG Avidity test. The IgG Avidity test proved a useful method to classify the Toxoplasma gondii infections in pregnancy, especially when we had only one serum sample.

HBV pre-vaccination screening in hospital personnel: cost-effectiveness analysis.

The purpose of this study was to identify the most cost-effective method for screening subjects for hepatitis B vaccination. Such a method would ideally permit detection of all susceptible individuals at the lowest possible cost. Two-hundred-five hospital workers from the Piedmont region of Italy participated in the study. The sero-epidemiological conditions of this group with regard to hepatitis B markers was representative of hospital workers in this region as a whole. All subjects, excluding carriers, persons with anti-HBs titers greater than or equal to 10 mIU and subjects positive for anti-HBc at a 1/100 dilution, were vaccinated. Their responses were evaluated 15 days and 1 month after vaccination. The presence of a booster effect following vaccination was correlated with the immunological status of the subject at the time of pre-vaccination screening. In the light of the results obtained, 5 screening procedures and the procedure of vaccination without screening were evaluated. The most cost effective screening strategy proved to be that of sequential testing for anti-HBc, anti-HBs and finally HBsAg and vaccination of the following subjects: those who were negative for anti-HBc, those who were anti-HBc-positive with anti-HBs titers 10 less than mIU and those who were HBsAg negative.