Oleuropein-Specific-13-Glucosidase Activity Marks the Early Response of Olive Fruits （Olea europaea） to Mimed Insect Attack

Olive fruits are seriously deteriorated by pre and postharvest damage due to the attack of insects, such as Bactrocera olaea, which strongly alters the quality of olives. Defence response in olive fruits injured both by pathogens and by mechanical damages has been associated with the enzyme β-glucosidase, which specifically hydrolyses oleuropein, producing highly reactive aldehyde molecules. In situ detection of ~-glucosidase activity in olive fruit tissues following injury, which simulates Bactrocera oleae punctures, is reported. The assay was performed in two cultivars showing different degrees of susceptibilities to fly infestation. In both cultivars, the histochemical assay for β-glucosidase showed that within 20 min after the injury, a strong ~-glucosidase activity could be observed in the damaged tissues. Thereafter a progressive enzyme inactivation occurred starting from tissues around the boundary of the injury with decrease of the enzyme activity and stopped after 3 h. Whereas the mass of active cells reached a distance of （300±50） μm from the edge of the injury. Biochemical analyses showed that in extracts of the injured fruit, β-glucosidase activity rapidly increased within 20 min from injury, thereafter decreasing and reaching values comparable with those in intact fruits. Following puncture, the oleuropein contents did not change significantly in the high susceptibility cultivar, whereas it rapidly decreased in the cultivar showing low susceptibility. The results strongly suggest that olive fruits susceptible towards fly infestation could be related to the ability of the oleuropein-degrading-β-glucosidase to produce the highly reactive molecules in the damaged tissues. As a consequence of puncture, high level of peroxidase activity was detected. This feature also suggested that this enzyme could play a key role in the defence response against insect injuries.     

Feline leishmaniosis due to Leishmania infantum in Italy

A case of leishmaniosis in domestic cats (Felis catus domesticus) is described. The subject showed a nodular lesion on the eyelid. The diagnosis was achieved by serological, parasitological, and light and electron microscopic investigations. By molecular techniques the aetiological agent was identified as belonging to Leishmania infantum, the species implicated in human and canine leishmaniosis in southern Europe. A preliminary study on the prevalence of asymptomatic feline leishmaniosis, performed in the areas where the infected cat was identified, revealed a low seroprevalence of infection: only 1 (0.9%) of the 110 cat sera examined by indirect fluorescent antibody test was positive for anti-Leishmania antibodies. Because clinical signs in feline leishmaniosis are unspecific and similar to those observed in other diseases commonly found in this species, leishmaniosis must be added to the differential diagnosis by feline veterinary practitioners and adequate serologic and histopathologic investigations must be performed in endemic areas.     

Isolation of Malassezia species from healthy cats and cats with otitis

Lipid-dependent Malassezia species have recently been cultured from veterinary specimens. The identification of Malassezia species isolates from animals is important to clarify the epidemiology of these lipophilic yeasts. Malassezia species were cultured from the external ear canals of 63 out of 99 cats with otitis and 12 of 52 (23%) healthy control cats. The rate of isolation in affected animals versus controls was highly significant (P<0.01). Malassezia pachydermatis was isolated as a pure culture in 33 (45.2%) cats, associated with Malassezia globosa and Malassezia furfur in 20 (50%) and 17 (42.5%) animals, respectively. Three different species were isolated simultaneously in three cats (two cats with M pachydermatis, M globosa and M furfur, one subject with M pachydermatis, M furfur and Malassezia sympodialis). M globosa was isolated as the sole species in two animals. The present work confirms the presence of some lipid-dependent species of Malassezia in both healthy and otitic cats.     

Application of hexanal,E-2-hexenal,and hexyl acetate to improve the safety of fresh-sliced apples

The aims of this work were to evaluate the effects of different concentrations of hexanal, (E)-2-hexenal, hexyl acetate, and their mixtures on the fate of pathogenic species such as Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes inoculated in model systems as well as the antimicrobial activity against the target species of the chosen molecules when added to the packaging atmosphere of inoculated fresh-sliced apples. The result obtained in this work pointed out the potential use of compounds such as hexanal, (E)-2-hexenal, and hexyl acetate for both the extension of shelf life and an improvement of hygienic safety of "minimally processed foods". In fact, hexanal, (E)-2-hexenal, and hexyl acetate had a significant inhibitory effect against pathogen microorganisms frequently isolated from raw materials (E. coli, S. enteritidis, and L. monocytogenes) when inoculated in both model and real systems. In this last condition, these compounds, at the levels used (150, 150, and 20 ppm for hexanal, hexyl acetate, and (E)-2-hexenal, respectively), displayed a bactericide effect on L. monocytogenes and they exhibited significant extensions of lag phase of E. coli and S. enteritidis inoculated at levels of 10(4)-10(5) CFU/g.     

Evaluating the Absorption of Boron by Plants��A Potential Tool to Remediate Contaminated Sediments from Cecina River Basin in Italy

Assessment of native plants and laboratory-scale phytoextraction tests are fundamental and preliminary steps in checking the feasibility and practice of low-cost and low-impact phytoremediation. In this study, we investigated the absorption of B by plants as a tool to remove boron in sediments from different areas of the Cecina River basin in Tuscany, Italy. The investigation was performed analyzing total and available B fraction in sediment samples as well as the B content in different tissues of native plants colonizing the contaminated areas. In laboratory scale, a phytoextraction screening test was performed. Selected high biomass crops (Brassica juncea, Zea mays, and Helianthus annuus) were evaluated in the most contaminated sample in two consecutive growing cycles. Results from field survey showed no hyperaccumulator native plant was present in the investigated areas although, high accumulation levels were found in native species from Bulera dump (Rumex crispus??259 mg?kg^?1 and Poa spp??203 mg?kg^?1). Results from laboratory phytoextraction tests showed a higher ability of B. juncea which removed about 18.5 mg?B?kg^?1 sediment in after the two consecutive growing cycles, representing on the whole 45% of the initial available B fraction. The sediment characteristics affected by the phytoextraction processes were also discussed.     

Fecal streptococci recoveries in different marine areas

The usual media and procedures were followed to measure the concentration of fecal streptococci (MPN on Azide Dextrose and Ethyl Violet Azide broths, membrane filtration on m-Enterococcus, KF and Pfizer Selective Enterococcus agars and according to the mE procedure) in samples collected along two different marine areas. The results were evaluated on the basis of three parameters: total concentrations, number of enterococci-like colonies (namely colonies gram positive, catalase negative, coccus shaped) and rate of strictly named fecal streptococci. From the results it appears that the various media and procedures employed gave different yields and their capacity to measure fecal streptococci varies according to the origin of samples. The accompanying bacterial flora may play an important role on the selectivity of each technique to measure the fecal streptococci.     

Diversity of human copy number variation and multicopy genes

Copy number variants affect both disease and normal phenotypic variation, but those lying within heavily duplicated, highly identical sequence have been difficult to assay. By analyzing short-read mapping depth for 159 human genomes, we demonstrated accurate estimation of absolute copy number for duplications as small as 1.9 kilobase pairs, ranging from 0 to 48 copies. We identified 4.1 million "singly unique nucleotide" positions informative in distinguishing specific copies and used them to genotype the copy and content of specific paralogs within highly duplicated gene families. These data identify human-specific expansions in genes associated with brain development, reveal extensive population genetic diversity, and detect signatures consistent with gene conversion in the human species. Our approach makes ~1000 genes accessible to genetic studies of disease association.     

Stand biometry and leaf area distribution in an old olive grove at Andria, southern Italy

The objectives of this paper were (1) to provide general biometry data for an 80-year-old olive (Olea europea L., cv. Coratina) grove in Andria, southern Italy, and (2) to compare different methods for estimating leaf area distributions. Stand biometry was represented by a stocking density of 132 trees ha^?1, mean spacing of 8.7 m and mean social area (proportional to spacing and tree size) of about 76 m² per tree. Trunk total circumference averaged 110 cm and after subtraction of missing or dead parts of stems averaged 81 cm, projected area of crowns averaged 17.7 m² and the mean tree height was 4.9 m. Leaf distribution was evaluated using calibrated ground-based side photographs through image analysis and through using a simple canopy-layer model (considering hollow volume within tree crowns) and double-Gaussian curves. The mean leaf size was about 5 cm² (distributed in a log-normal manner over the range of 2 to 12 cm²). Considering whole tree crowns, the mean leaf density was about 2.6 m²m^?3; the maximum leaf area occurred in canopy layers between 1.5 to 3 m, tailing with a steeper slope to the crown base and a less steep slope to the tree-top. The foliated volume of olive crowns (mean 33.2 m³) contained on average 145 thousand leaves of the total area of 72.6 m². The corresponding leaf area index on the stand level (LAI _grove = 0.96), was rather low due to low stocking density. However when taking into account only the projected crown areas (and avoiding free space between trees), the mean LAI reached about 3.5 (range from 1–7). The radial pattern of leaf distribution derived from image analysis indicated peak LAI _rad values at a distance from the stem of about 60 to 70% of crown radius in trees of different size. The applicability of different approaches to the estimation of the necessary allometric parameters is discussed.     

Survey of polychlorinated dibenzo-p-dioxins (PCDDs), Polychlorinated Dibenzo-p-furans (PCDFs), polychlorinated biphenyls (PCBs), and mineral components in Italian citrus cold-pressed essential oils

Investigation of PCDDs, PCDFs, PCBs, Al, As, Pb, Ba, Co, Cu, Cr, Fe, Mn, Cd, Ag, Sn, Zn, and Hg contents in 60 samples of cold-pressed essential oils produced in Calabria and Sicily in 2003-2005 was carried out. PCDDs, PCDFs, and PCBs were analyzed by HRGC-HRMS techniques using U.S. EPA 1613/94 and U.S. EPA 1668/A (1999) analytical methods. Mineral components were determined through GFAAS techniques; Hg content was determined by FI-M/H-AAS. The results of this study showed that essential oil contamination was due to a widespread pollution, typical background of rural areas, with relatively higher concentrations of PCDDs compared to PCDFs and little presence of PeCDF. Congeners OCDD, HpCDF, and OCDF were found at high concentrations. Regarding mineral components, mean values of Cr, Fe, and Ni were in agreement with data reported in the literature. Concentrations of As and Pb were below the maximum limits accepted by the current legislation. Finally, none of the samples analyzed were contaminated with Hg.     

Identification of PCR-amplified genetically modified organisms (GMOs) DNA by peptide nucleic acid (PNA) probes in anion-exchange chromatographic analysis

PCR products obtained by selective amplification of transgenic DNA derived from food samples containing Roundup Ready soybean or Bt-176 maize have been analyzed by anion-exchange HPLC. Peptide nucleic acids (PNAs), oligonucleotide analogues known to bind to complementary single-stranded DNA with high affinity and specificity, have been used as specific probes in order to assess the identity of the peaks observed. Two different protocols were adopted in order to obtain single-stranded DNA: amplification with an excess of one primer or digestion of one DNA strand. The single-stranded DNA was mixed with the PNA probe, and the presence of a specific sequence was revealed through detection of the corresponding PNA:DNA peak with significantly different retention time. Advantages and limits of this approach are discussed. The method was tested with reference materials and subsequently applied to commercial samples.