Role of secondary metabolites in the biocontrol activity of <Emphasis Type="Italic">Pseudomonas corrugata</Emphasis> and <Emphasis Type="Italic">Pseudomonas mediterranea</Emphasis>

Colletotrichum gloeosporioides is the causal agent of Camellia oleifera anthracnose, mainly infecting fruits and leaves. The fungus secretes degrading enzymes to destroy the cuticle of aerial plant parts and help infect the host successfully. To validate whether a cutinase gene (CglCUT1) was required for cutinase activity and pathogenicity of C. gloeosporioides, the CglCUT1 gene was cloned and analyzed. The characterization of CglCUT1 predicted protein suggests that the cloned DNA encoded a cutinase in C. gloeosporioides affecting C. oleifera. The CglCUT1 showed a high homology to those from C. gloeosporioides causing papaya anthracnose and C. capsici causing pepper anthracnose, as well as those of other ascomycetes. The whole CglCUT1 gene was knocked-out and the knockout mutant (?CglCUT39) was subsequently complemented using Agrobacterium tumefaciens mediated transformation. The knockout transformants exhibited significant decreases in cutinase activity and virulence compared with the wild-type strain. The complemented transformants of the disrupted transformant ?CglCUT39 showed a significant increase in cutinase activity and virulence compared with the disrupted transformant ?CglCUT39. This study suggests that the CglCUT1 gene has a positive effect on fungal virulence of the hemibiotrophic C. gloeosporioides on C. oleifera.     

Identification of virgin olive oil from different cultivars by analysis of DNA microsatellites

DNA analysis enables genome fingerprinting with consequent identification of different individuals. In the agro-food industry, this can have interesting applications for the identification of species and cultivars of both raw materials and processed food. In this investigation, the efficiency of DNA microsatellite analysis in identifying virgin olive oils from different cultivars was evaluated. Ten virgin oils were obtained in the laboratory from olives of 10 different cultivars and the DNA extracted from the cell residues, recovered by oil centrifugation, was used as a template with seven different primer pairs of microsatellite sequences. The electrophoretic patterns showed an adequate level of amplification and were identical to those obtained from leaves and drupes of the same cultivar. By analyzing all the patterns obtained, the smallest number of microsatellites able to distinguish the examined oils was established and an identification key for the different oils was developed.     

Flumequine Uptake and the Aquatic Duckweed,Lemna minor L.

Water, Air, & Soil Pollution - Phytotoxicity of Flumequine (F) on the aquatic duckweed, Lemna minor L., and plant drug uptake were evaluated by a simple ecotoxicological test. Flumequine, at...     

Thermal degradation studies of food melanoidins

Food melanoidins were isolated from bread crust, coffee, and tomato sauce and their composition was investigated by thermal degradation. Among the generated volatiles, important food flavor compounds were detected: in particular furans, carbonyl compounds, 1,3-dioxolanes, pyrroles, pyrazines, pyridines, thiophenes, and phenols. The results indicated that the isolated melanoidin fractions mainly consisted of compounds formed from carbohydrates and their degradation products. Besides proteins, other food constituents were incorporated in the melanoidin structure as well, such as lipid oxidation products in tomato melanoidins and phenolic compounds in coffee melanoidins. A comparison of the thermal generation of volatiles between these food-derived melanoidins and model melanoidins prepared from a single carbonyl compound and an amino acid showed that the degradation pattern of food melanoidins is quite different from that obtained from a glucose-glycine model system.     

Classification of Italian honeys by 2D HR-NMR

The importance of honey has been recently increased because of its nutrient and therapeutic effects, but the adulteration of honey in terms of botanical origin has increased, too. The floral origin of honeys is usually determined using melisso-palynological analysis and organoleptic characteristics, but the application of these techniques requires some expertise. A number of papers have confirmed the possibility of characterizing honey samples by selected chemical parameters. In this study high-resolution nuclear magnetic resonance (HR-NMR) and multivariate statistical analysis methods were used to identify and classify honeys of five different floral sources. The 71 honey samples (robinia, chestnut, citrus, eucalyptus, polyfloral) were analyzed by HR-NMR using both 1H NMR and heteronuclear multiple bond correlation spectroscopy (HMBC). Spectral data were analyzed by application of unsupervised and supervised pattern recognition and multivariate statistical techniques such as principal component analysis (PCA) and general discriminant analysis (GDA). The use of 1H-(13)C HMBC coupled with appropriate statistical analysis seems to be an efficient technique for the classification of honeys.     

Environmental influences on the recruitment dynamics of juvenile European eels,Anguilla anguilla,in a small estuary of the Tyrrhenian Sea,Sardinia, Italy

1. The European eel, Anguilla anguilla, is a catadromous and migratory species of commercial importance. Its complex life cycle results in its exposure to many risk factors, which have resulted in stock declines across all life stages since the 1970s.
2. The temporal recruitment dynamics of juvenile eels (glass eels and elvers) were investigated in a small Mediterranean estuary (Sardinia, Italy). The composition of the population and the monthly and seasonal variations in the abundances of juvenile eels was assessed over 78 sampling events (from February 2017 to February 2018). Furthermore, the effects of abiotic variables on the abundances of glass eels and elvers were investigated using generalized additive models (GAMs).
3. Glass eels had the greatest abundance during the winter months, whereas elvers had the greatest abundance during spring. Modelling revealed that the abundance of glass eels was mostly explained by the combined effects of water temperature (12.3–14.5 °C), tidal coefficient (40–110 cm), moon phase, season, and river mouth condition, whereas the abundance of elvers was associated with water temperature (14–21 °C), dissolved oxygen content (>7 mg/L), and season. These results suggest that the annual recruitment of juvenile eels occurs throughout the year, with clear seasonal migration dynamics.
4. The use of multiple statistical approaches allowed us to identify the importance of several environmental variables in regulating the recruitment dynamics, providing useful information for conserving eel stocks through the restoration of the natural flow regime and the connectivity between freshwater habitats and the sea.

     

Botryoid nuclei in the peripheral blood of a dog with heatstroke

An EDTA‐anticoagulated blood sample collected from a 1.5‐year‐old, intact male, English Bulldog was submitted for a CBC. The CBC data and blood smear evaluation revealed borderline high hematocrit (54%, reference interval 37–55%), inappropriate rubricytosis, moderate leukopenia due to both mature neutropenia and lymphopenia, and mild thrombocytopenia. Numerous leukocytes showed evidence of karyolysis, pyknosis, and karyorhexis, and apoptotic bodies were frequent in the background. Many neutrophils had botryoid nuclei characterized by increased numbers of nuclear segments radially arranged with spoke‐like, delicate chromatin filaments connecting the segments centrally. The finding of botryoid nuclei and inappropriate rubricytosis was indicative of severe hyperthermia, such as heatstroke. The dog had been exercised a long time during conditions of high temperature and humidity until he collapsed. The dog was diagnosed with severe heatstroke, hypovolemic shock, disseminated intravascular coagulation, and multiorgan dysfunction syndrome. Despite aggressive treatment, the patient died of cardiopulmonary arrest. Botryoid nuclei are frequent in people with heatstroke. In the authors' experience, botryoid nuclei are seen commonly in dogs with heatstroke, but they have never been reported in veterinary medicine. The presence of petechiation with only mild thrombocytopenia and inappropriate rubricytosis also is suggestive of heatstroke and manifests ongoing life‐threatening vascular derangement.     

Amplified fragment length polymorphism for variety identification and genetic diversity assessment in oleander (Nerium oleander L.)

Oleander is a Mediterranean evergreen shrub found along watercourses, gravelly places and damp slopes. It is grown widely as an ornamental for its abundant and long-lasting flowering as well as its moderate hardiness. Genetic relatedness among 71 accessions, including commercial varieties, different sources of the same varieties, and selections from the wild were investigated using amplified fragment length polymorphism (AFLP). Nine primer combinations yielded a total of 603 bands of which 241 were polymorphic. Genetic similarities among accessions were calculated according to Jaccard's Similarity Index and used to construct a dendrogram based on the unweighted pair group method using arithmetic averages. Our results show that the AFLP technique, which can simultaneously and assay a large number of loci randomly distributed in the genome, is much more informative on the genetic relationship and origin of accessions than the limited number of morphological characters conventionally used for variety discrimination. Up to about 9% molecular genetic differentiation was detected among morphologically indistinguishable provenances of the same variety; this can be partly attributed to scoring error but mainly to somatic variation occurring during vegetative propagation. On the other hand lower genetic distance values were sometimes found among varieties which differ in morphological characters and are thus commercialised with different names. The possibility of considering the amount of genetic variation within a variety as the threshold value for discrimination of initial varieties and essentially derived varieties is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Intra-cultivar variability of three major olive cultivars grown in different areas of central-southern Italy and studied using microsatellite markers

A collection of 70 olive samples, originating from diverse areas in central-southern Italy (Abruzzo, Apulia, Calabria, and Umbria) and corresponding to 3 major cultivars denominations (‘Carolea’, ‘Coratina’ and ‘Frantoio’), was genotyped at 10 microsatellite loci. In total, 44 alleles with a mean number of 4.4 alleles per locus were detected. The molecular analysis, allowed the study to show a clear genetic diversity between the three cultivars ‘Carolea’, ‘Coratina’ and ‘Frantoio’ and to state that ‘Carolea’ is a polyclonal cultivar, while ‘Coratina’ and ‘Frantoio’, are probably monoclonal ones. The analysis of intra-varietal polymorphism, through the SSR analysis, proved to be very useful both for varietal identification and for intra-varietal ones. Our work shows that the current designations of olive cultivars fall short of describing the genetic variability among economically important plant material. A thorough investigation of the existing variability will prove of major importance for both management and economic production of olive trees.