Francesca Cibelli

Phaeoacremonium species associated with olive wilt and decline in southern Italy

Six Phaeoacremonium species (spp.) were isolated from symptomatic wood of olive trees (Olea europea) in Apulia (southern Italy) that showed crown wilt and twig and branch dieback. These Phaeoacremonium spp. were identified according to their morphological characteristics and by analyses of partial sequences of the actin and β-tubulin genes. Combining these cultural, morphological and molecular data, three Phaeoacremonium spp. were isolated that are already known to be responsible for severe decline of olive in Apulia, Phaeoacremonium aleophilum, Phaeoacremonium alvesii and Phaeoacremonium parasiticum, together with three other Phaeoacremonium spp. that are associated for the first time with wilt, decline and dieback of olive orchards in Italy and worldwide: Phaeoacremonium italicum, Phaeoacremonium sicilianum and Phaeoacremonium scolyti. To understand and to confirm their involvement in wilt and decline of olive trees, pathogenicity assays were performed on shoots of young olive plants. The data indicate that all six of these Phaeoacremonium spp. can cause discolouration, necrotic wood, and death of shoots, although different levels of virulence were observed, with Pm. italicum, Pm. aleophilum and Pm. sicilianum producing greater necrotic lesions than the other Phaeoacremonium spp. investigated here.

Metabiotic Effects of Fusarium spp. on Escherichia coli O157:H7 and Listeria monocytogenes on Raw Portioned Tomatoes

The metabiotic effects of Fusarium proliferatum, F. avenaceum, and F. oxysporum on Escherichia coli O157:H7 and Listeria monocytogenes in fresh tomatoes were investigated. Tomatoes were preinoculated with the molds and incubated at 15 degrees C for 7 days; then they were inoculated separately with the pathogens, packaged in air and modified atmosphere (5% O2, 30% CO2, and 65% N2), and stored at 4, 8, and 12 degrees C for 9 days. The cell loads of pathogens and lactic acid bacteria and the pH were evaluated periodically. The data were modeled through some different mathematical models to assess the shoulder length, i.e., the time before the beginning of the exponential death phase, the 1-log reduction time (s), and the pathogen death time (deltastand). The preinoculation of tomatoes with the molds enhanced the survival of E. coli O157:H7 by prolonging shoulder length and 8 parameters; this effect, however, was not observed for L. monocytogenes. pH values did not undergo significant changes within the storage time, and the lactic acid bacteria increased from 5 to 7 log CFU/g, without significant differences among the storage temperatures or the packaging atmospheres. The results of this research showed that the use of fresh tomatoes colonized by fusaria (even if the contamination is not visible) could increase significantly the risk of outbreaks due to some pathogens that could be on the surface of fruits and vegetables as a result of cross-contamination at home or incorrect postharvest operations.

Fungal bioremediation of olive mill wastewater: Using a multi-step approach to model inhibition or stimulation.

BACKGROUND Olive mill wastewaters (OMWWs) possess a strong environmental impact; the use of fungi as tools for bioremediation could be a promising method. RESULTS Twenty-nine fungi were grown on minimal media supplemented with five different kinds of OMWWs (5-15%). Radial growth was assessed for 21 days and the data were modelled through the Dantigny-logistic like function to estimate τ, i.e. the time to attain half of the maximum diameter. Growth on potato dextrose agar and water agar (WA, minimal medium without supplementation) was used as reference. The differences in τ between PDA/WA and minimal media with OMWWs were modelled through a multi-factorial ANOVA, using the concentration of OMWW, the kind of wastes and fungi as categorical predictors. Finally, a principal component analysis was run to group and divide resistant and sensitive fungi. Some fungi experienced a positive Δτ, thus suggesting an inhibition by OMWW, whereas other isolates were enhanced. CONCLUSIONS Some isolates (for example Aspergillus ochraceus) showed a promising trend and could be possible candidates for a validation on a real scale.

Proteolytic activity of molds and their metabiotic association with Salmonella in a model system.

The aim of this study was to investigate the proteolytic ability of some strains of aspergilli, fusaria, and penicillia and the metabiotic effect of Fusarium oxysporum and Penicillium expansum on Salmonella. The proteolytic activity of the target molds was determined on tomato juice agar and tomato juice, whereas the metabiotic effect of F. oxysporum and P. expansum on Salmonella was assessed in a model system consisting of tryptone soy broth with different amounts of tomato juice added. Fusaria, some aspergilli, and one strain of penicillium showed a proteolytic activity on tomato juice agar. In addition, Salmonella survival was enhanced in tryptone soy broth plus 20 or 50% tomato juice in the model system previously inoculated with F. oxysporum.

First report of stem wilt and root rot of Schlumbergera truncata caused by Fusarium oxysporum f. sp. Opuntiarum in southern Italy.

Schlumbergera truncata (Haw.) Moran, belonging to the Cactaceae, is a very common ornamental cactus in southern Italy. In November 2011, sudden stem wilt and root rot was observed in about 45% of vegetatively propagated plants cultivated as potted ornamental plants in a commercial greenhouse in Cerignola (Foggia Province, Apulia, Italy). The roots and collars of the plants showed brown rot. Yellow sunken lesions that were similar to cortical cankers were detected at basal level of the stem. Ten plants with these symptoms were analyzed by fungal isolation techniques. Small (0.5 cm) tissue portions from root, collar, and basal stem were plated on potato dextrose agar (PDA) after disinfection with 75% ethanol for 1 to 2 min, 0.2% NaOCl for 1 to 2 min, and a wash with sterile distilled water. A fungal isolate that was morphologically similar to Fusarium sp. was isolated from 85% of these tissue samples. It had nucleotide sequences of the internal transcribed spacer region (ITS1-5.8S-ITS2) of ribosomal DNA (GenBank Accession No. KC196121) 100% identical to those of the comparable sequences of Fusarium oxysporum (HQ651161). The nucleotide sequences of its translation elongation factor 1-α (EF-1α) gene (KC196120) showed 100% identity to sequences of F. oxysporum f. sp. opuntiarum (DQ837689, AF246881) retrieved from GenBank. Pathogenicity tests were performed at 22 ± 3°C on 18 45-day-old plants of S. truncate by adding of a 5-ml aliquot of conidial suspension adjusted to 5 × 106 conidia/ml to soil of each plant. Six non-inoculated plants were used for a control treatment and sprayed with 5 ml of sterilized water. Plants were maintained in greenhouse at 22 ± 3°C. After 10 days, nine of the inoculated plants showed wilting, and after 45 days, all of them were dead, with root and collar rot and lesions on the basal stem. Control plants were symptomless. Kochs postulates were fulfilled as the pathogen was reisolated from all of the symptomatic tissues and identified as Fusarium sp. On the basis of 3-septate macroconidia (mean 31.75 × 3.21 μm; range, 26 to 35 μm long, 3.0 to 4.2 μm wide), aseptate microconidia, single chlamydospores, and monophialide conidiophores on carnation leaf agar, and molecular analyses, the fungus was identified as F. oxysporum f. sp. opuntiarum (Speg) (1,2,3). In Italy, F. oxysporum f. sp. opuntiarum was reported as basal stem rot of Echinocactus grusoni (4). To our knowledge, this is the first report of stem wilt and root rot of S. truncata caused by F. oxysporum f. sp. opuntiarum in Italy. References: (1) W. Gerlach. Phytopathol. Z. 74:197, 1972. (2) W. L. Gordon. Can. J. Bot. 43:1309, 1965. (3) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University Press, University Park, 1983. (4) G. Polizzi et al. Plant Dis. 88:85, 2004.

Evaluation of Fungal Growth on Olive-Mill Wastewaters Treated at High Temperature and by High-Pressure Homogenization

Reuse of olive mill wastewaters (OMWWs) in agriculture represents a significant challenge for health and safety of our planet. Phytotoxic compounds in OMWW generally prohibit use of untreated OMWWs for agricultural irrigation or direct discharge into surface waters. However, pretreated OMWW can have positive effects on chemical and microbiological soil characteristics, to fight against fungal soil-borne pathogens. Low amounts of OMWW following thermal (TT-OMWW) and high-pressure homogenization (HPH-OMWW) pretreatments counteracted growth of some of 12 soil-borne and/or pathogenic fungi examined. With fungal growth measured as standardized change in time to half maximum colony diameter, Δτ, overall, HPH-OMWW showed increased bioactivity, as increased mean Δτ from 3.0 to 4.8 days. Principal component analysis highlighted two fungal groups: Colletotrichum gloeosporioides, Alternaria alternata, Sclerotium rolfsii, and Rosellinia necatrix, with growth strongly inhibited by the treated OMWWs; and Aspergillus ochraceus and Phaeoacremonium parasiticum, with stimulated growth by the treated OMWWs. As a non-thermal treatment, HPH-OMWW generally shows improved positive effects, which potentially arise from preservation of the phenols.

Modelling the Survival of Escherichia coli O157:H7 on Raw Portioned Tomatoes, Inoculated with Aspergillus fumigatus and Emericella nidulans

The metabiotic interactions occurring among two fungi (Aspergillus fumigatus and Emericella nidulans) and Escherichia coli O157:H7 on raw portioned tomatoes were studied. Tomatoes, preinoculated with the moulds and inoculated with the pathogen, were packaged in air and stored at 4, 8 and 12∘C for 9 days; pathogen cell number and pH were monitored throughout the storage and the data were modeled using three different equations (Geeraerd, Weibull, and modified Weibull), to assess the shoulder length, the 1-log reduction time, and the death time. Both A. fumigatus and E. nidulans increased the survival of E. coli O157:H7 through the prolongation of the shoulder length; in contrast, the death time was significantly increased. The results of this paper suggested that the metabiotic interactions aspergilli/E. coli O 157:H7 could be of public concern, as the consumption of tomatoes (or other fruits and vegetables) contaminated both by the moulds and the pathogen is a possible scenario.