Root and stem rot,and wilting of olive tree caused by Dematophora necatrix and associated with Emmia lacerata in Central Italy

European Journal of Plant Pathology - Lethal wilting was observed on young olive trees cv Favolosa in a grove in central Italy. White mycelial strands wrapped the basal portion of the stems that...     

Effect of Metarhizium anisopliae fungus on off-host Rhipicephalus (Boophilus) microplus from tick-infested pasture under cattle grazing in Brazil

This work aimed to assess the effect of the fungus Metarhizium anisopliae on off-host Rhipicephalus (Boophilus) microplus from tick-infested Brachiaria decumbens pasture undergoing cattle grazing. For this purpose a naturally tick-infested Brachiaria decumbens pasture of 60 m × 100 m with twelve grazing Holstein Friesian-Nelore (Bos indicus) cross breed bovines was sprayed 12 times, 21 days apart with an aqueous conidial suspension of the E9 isolate of M. anisopliae fungus. Control pasture was treated with conidial suspension vehicle only. Efficacy of treatment was evaluated by tick larvae counts on the pasture and that of engorged female ticks on grazing cattle. Fungus persistence on grass stems as well as soil pasture was assessed after each spray. Efficacy of fungus against ticks was also measured by an in vitro immersion test. Whereas in vitro test showed a clear pathogenic effect of the fungus on ticks, no pathogenic effect over R. (B.) microplus ticks was detected in the field trial by spraying pasture with fungal suspension. Fungus from the suspension could be recovered from both the grass stem as well as the soil of sprayed pasture, even though the numbers obtained varied distinctly and could only be recovered shortly after spray. Pasture environments with exposure to sun and rain, seem to be very detrimental to the fungus, thus suspensions which provide fungus with protection or more resistant isolates to these should be looked for. In order to achieve high fungal efficacy against tick under field conditions, accurate laboratory assays, optimization of application strategy and knowledge of interactions between fungal strains and environmental factors are warranted.     

Tetrodotoxins (TTXs) and Vibrio alginolyticus in Mussels from Central Adriatic Sea (Italy): Are They Closely Related?

Tetrodotoxins (TTXs), potent neurotoxins, have become an increasing concern in Europe in recent decades, especially because of their presence in mollusks. The European Food Safety Authority published a Scientific Opinion setting a recommended threshold for TTX in mollusks of 44 µg equivalent kg⁻¹ and calling all member states to contribute to an effort to gather data in order to produce a more exhaustive risk assessment. The objective of this work was to assess TTX levels in wild and farmed mussels (Mytilus galloprovincialis) harvested in 2018–2019 along the coastal area of the Marche region in the Central Adriatic Sea (Italy). The presence of Vibrio spp. carrying the non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) genes, which are suspected to be involved in TTX biosynthesis, was also investigated. Out of 158 mussel samples analyzed by hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry (HILIC-MS/MS), 11 (7%) contained the toxins at detectable levels (8–26 µg kg⁻¹) and 3 (2%) contained levels above the EFSA safety threshold (61–76 µg kg⁻¹). Contaminated mussels were all harvested from natural beds in spring or summer. Of the 2019 samples, 70% of them contained V. alginolyticus strains with the NRPS and/or PKS genes. None of the strains containing NRPS and/or PKS genes showed detectable levels of TTXs. TTXs in mussels are not yet a threat in the Marche region nor in Europe, but further investigations are surely needed.     

Antibodies to soluble liver antigen and α-enolase in patients with autoimmune hepatitis

BACKGROUND: Antibodies to a cytosolic soluble liver antigen (SLA) are specifically detected in patients with autoimmune hepatitis (AIH). The target of anti-SLA has been identified as a ~50 kDa UGA serine tRNA-associated protein complex (tRNP(Ser)Sec), through the screening of cDNA libraries. A recent report questioned the identity of tRNP(Ser)Sec as the real SLA antigen. The latter study identified alpha-enolase as a major anti-SLA target, through proteomic analysis. METHODS: In an attempt to explain the observed discrepancy we have investigated reactivity of SLA positive sera against alpha-enolase and tRNP(Ser)Sec using rat and primate liver homogenate and the recombinant antigens. Thirty-three serum samples, 11 from SLA-positive patients and 22 from SLA negative controls were investigated. SLA antibodies were detected by an inhibition ELISA and confirmed by immunoblot using human liver homogenate. Autoantibody reactivity was further evaluated using preparations of primate and rat liver homogenates. Anti-alpha-enolase antibody reactivity has been tested by immunoblot using recombinant alpha-enolase. An affinity purified goat polyclonal anti-alpha-enolase IgG antibody was used as reference serum sample. Anti-tRNP(Ser)Sec antibody reactivity was detected by ELISA or dot blot using recombinant tRNP(Ser)Sec antigen. RESULTS AND DISCUSSION: The affinity purified IgG antibody directed to human alpha-enolase gave a band of approximately 48 kDa in both human and rat liver homogenates. A high titre anti-tRNP(Ser)Sec antibody serum gave a single band of ~50 kDa in both liver preparations. All but one anti-SLA antibody positive sera reacted with a ~50 kDa but none immunofixed a 48 kDa band. All anti-SLA antibody positive sera reacted strongly with the recombinant full length tRNP(Ser)Sec protein. None of the anti-SLA negative sera reacted with tRNP(Ser)Sec. Anti-SLA positive, and anti-SLA negative sera reacted equally against recombinant alpha-enolase by immunoblot. Pre-incubation of anti-SLA positive sera with tRNP(Ser)Sec completely abolished the 50 kDa band. The findings of the present study indicate that alpha-enolase and tRNP(Ser)Sec are both expressed in primate and rat liver and have a respective MW of 48 and 50 kDa. They also show that anti-tRNP(Ser)Sec - but not anti-alpha-enolase - correlates with anti-SLA antibody reactivity. CONCLUSION: Our findings indicate that tRNP(Ser)Sec is the most likely target of anti-SLA.     

Collection time,cutting age,IBA and putrescine effects on root formation in Corylus avellana L. cuttings

The effects of collection time, cutting age, indole-3-butyric acid (IBA) and putrescine application on the rooting of cuttings of Italian hazelnut cultivars ‘Tonda Gentile Romana’, ‘Tonda di Giffoni’ and ‘Nocchione’ were investigated. Samples collected during late June, late July and early September from newly formed and 1-year old part of twigs to be utilized to produce leafy cuttings, after being treated with 1000 and 2000 ppm IBA, respectively. In addition, the September cuttings were also treated, respectively, with 1000 and 2000 ppm IBA and 1600 ppm putrescine. Rooting ability was evaluated 2 months after planting for each treatment and collection time. Satisfactory rooting of hazelnut leafy cuttings was observed when collection time occurred in June and September, whereas leafy cuttings collected in July showed a limited capacity of rooting in all cultivars tested. On average, the rooting of the newly formed leafy cuttings was more than the 1-year old cuttings. Rooting was also promoted by IBA treatments, mainly in ‘Nocchione’ and ‘Tonda di Giffoni’. In contrast, young cuttings collected from ‘Tonda Gentile Romana’ in early September rooted poorly when treated with IBA alone, but showed the best rooting (∼80%) after the application of a combination of 1000 ppm IBA and 1600 ppm putrescine. The current findings confirm that putrescine can be a useful substance for increasing rooting percentage and root quality in cuttings of some hazelnut cultivars as obtained from ‘Tonda Gentile Romana’ cultivar.     

How is agroforestry perceived in Europe? An assessment of positive and negative aspects by stakeholders

Whilst the benefits of agroforestry are widely recognised in tropical latitudes few studies have assessed how agroforestry is perceived in temperate latitudes. This study evaluates how stakeholders and key actors including farmers, landowners, agricultural advisors, researchers and environmentalists perceive the implementation and expansion of agroforestry in Europe. Meetings were held with 30 stakeholder groups covering different agroforestry systems in 2014 in eleven EU countries (Denmark, France, Germany, Greece, Hungary, Italy, Netherlands, Portugal, Spain, Sweden and the United Kingdom). In total 344 valid responses were received to a questionnaire where stakeholders were asked to rank the positive and negative aspects of implementing agroforestry in their region. Improved biodiversity and wildlife habitats, animal health and welfare, and landscape aesthetics were seen as the main positive aspects of agroforestry. By contrast, increased labour, complexity of work, management costs and administrative burden were seen as the most important negative aspects. Overall, improving the environmental value of agriculture was seen as the main benefit of agroforestry, whilst management and socio-economic issues were seen as the greatest barriers. The great variability in the opportunities and barriers of the systems suggests enhanced adoption of agroforestry across Europe will be most likely to occur with specific initiatives for each type of system.     

Separation of pesticides,related compounds,polychlorobiphenyls and other pollutants into four groups by silica-gel microcolumn chromatography (application to surface water analysis)

Multi-residue analyses of pesticides and related compounds generally require clean-up procedures for the elimination of interfering substances. An analytical procedure for use in the recovery and determination of pesticides and other pollutants in water samples is described, in which the purification of the extracts is improved by the separation of pesticides into four groups. The separation of pesticides by silica-gel microcolumn chromatography is explained in detail.     

Cryopreservation of European Mouflon (Ovis Gmelini Musimon) Semen During the non-Breeding Season is Enhanced by the Use of Trehalose

The influence of trehalose on European mouflon spermatozoa cryopreservation during the non-breeding season was tested. Semen was frozen in two different extenders: (a) recommended Tris-based ram extender (CTR); (b) CTR extender supplemented with trehalose 0.147 mm (TRH). Sperm viability and acrosome integrity were assessed using propidium iodide and fluorescein isothiocynate labelled Pisum Sativum agglutinin. Trehalose significantly enhanced sperm viability after thawing compared with CTR extender (62.7% vs 51.8%; p < 0.05), whereas no differences were observed on acrosome integrity (42.9% vs 42.1%). Trehalose influence was also evidenced in the in vitro fertility test performed with sheep oocytes matured in vitro. Both fertilization rates (60.9% TRH vs 43.6% CTR; p < 0.05) and cleavage rates (58% TRH vs 39.8% CTR; p < 0.001) were higher for trehalose frozen semen compared with control extender frozen semen. A higher percentage of zygotes resulting from fertilization with trehalose cryopreserved semen presented the first cleavage earlier if compared with the group fertilized with control semen (48.7% vs 31.5%, respectively; p < 0.01). This result was confirmed by embryo kinetic development. Fertilization with trehalose cryopreserved semen leaded to an higher percentage of blastocysts (40.2% vs 27.8% CTR; p < 0.05), and enhanced in particular the number of blastocysts that developed on the day 6th of culture (28.6% vs 17% CTR; p < 0.05). Our data demonstrated that, during mouflon non-breeding season, trehalose extender enhances spermatozoa viability and its in vitro fertilizing capacity, allowing the production of an higher number of blastocysts.