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.     

Oleuropein-Specific-β-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 β-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 β-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 3h. Whereas the mass of active celsreached 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.     

Molecular phylogeny of the apterygotan insects based on nuclear and mitochondrial genes

Basal hexapodan orders (Apterygota) are traditionally divided in two well defined taxonomic groups, Entognatha and Ectognatha. Entognathy (the enclosed mouthparts condition) is considered the most distinctive character joining the Ellipura (Protura+ Collembola) with the order Diplura, whereas the presence of exposed mouthparts (ectognathy) is the feature shared by Microcoryphia and Zygentoma with the pterygote insects. In spite of the growing interest for the evolutionary history of the Apterygota, there is no complete agreement among the general phylogenetic hypotheses based on the study of morphological characters. In this study we analyzed the DNA sequence of segments of the nuclear Elongation Factor-1α(EF-1α) and of the mitochondrial 12S rRNA genes, and used different methods of phylogenetic reconstruction. The 12S gene seems to be more suited than the EF-1α to resolve some of the most outstanding systematic disputes, whereas the lack of resolution at the deeper nodes does not allow to assess the phylogenetic relationships within Microcoryphia and between ectognathan orders. We have obtained a fairly high support for the monophyly of the orders Diplura and Zygentoma. In the 12S analysis, the Ellipura and the Entognatha form monophyletic assemblages. In addition, the study of the distribution of introns in the EF-1α suggests a relationship between Collembola and Diplura.     

Re-shaping the “original SIN”: a need to re-think sediment management and policy by introducing the “buffer zone” concept

Purpose

The effects of long-standing industrialization processes and poor environmental management practices have often left a harmful legacy for marine-coastal sites worldwide, causing a wide range of unforeseen impacts on the ecosystem and on human health. A critical revision of available data from three highly contaminated Italian sites (Augusta Bay, Sicily; Cagliari Gulf, Sardinia; Pozzuoli Bay, Campania) revealed the crucial role of biogeochemical/physical dynamics and potential widespread delivery of contaminants as key components for a wider comprehensive sediment management.

Materials and methods

Datasets of organic pollutants (ΣPAHs, ΣPCBs), heavy metals (As, Cd, Co, Cr, Cu, Hg, Ni, Pb, V, and Zn), radionuclides, and grain size composition of marine sediments of all three sites were critically reviewed and seafloor bathymetric data were analyzed.

Results and discussion

A critical review of available physical and chemical information of sediments collected from the three marine sites provides a better understanding of the physical and biogeochemical dynamics of large-scale dispersion of contaminants in those areas, as well as new insights in terms of improved sediments management policies with actions of larger scale monitoring of the “buffer zone” confining with the national relevance and highly contaminated site to trace inputs and accumulation effects of pollutants deriving from the point-source.

Conclusions

This study demonstrates how the presence of polluted sediments on the coastal areas can have, also after the closure of anthropogenic activities, a potential impact at a large scale. Then to better evaluate the effective impact, we suggest the adoption of a buffer zone contiguous to the national relevance sites (SIN) area. While SINs are optimized to define the state of the art of the near-source pollution, buffer zones may help to outline the rate of delivery of pollutants to the deep sea. Buffer zones must be designed by taking into account the local hydrodynamics, sedimentology, and geomorphology, and should be monitored, even if at a lower resolution, like SINs.

     

p62/SQSTM1 expression in canine mammary tumours: Evolutionary notes

Recent studies highlighted the role of autophagy as a cardinal regulatory system for homeostasis and cancer‐related signalling pathways. In this context, the deregulated expression of p62 – Sequestosome1 (p62/SQSTM1) – a protein acting both as an autophagy receptor and signalling hub, has been associated with tumour development and chronic inflammation. Multiple clinical studies test drugs targeting autophagy, and even more research is on the way to clinical trials. However, no comparative investigations have been carried out to identify adequate preclinical models to assess p62‐based medicine. In veterinary oncology the role of p62 in cancer‐related pathways has been largely ignored. We compared p62 sequences in multiple organisms and found that canine p62 significantly diverges from the humans and from other animals sequences. Then, we chart by immunohistochemistry the expression levels of p62 in canine mammary tumours. A total of 66 tumours and 10 non‐neoplastic mammary samples were examined. The expression of p62 was higher in normal tissue and adenomas than carcinomas, with lowest levels of p62 protein detected in high grade carcinomas. In all cases examined the tumour stroma appeared to be p62‐negative. Taken together our results would suggest that in dogs the association between p62 expression and cancer cells overturns that reported in human breast carcinoma, where p62 accumulates in malignant cells as compared to normal epithelium. Thus, at least in canine mammary tumours, p62 should be not considered a tumour‐rejection antigen for an anti‐cancer immunotherapy.     

Immunological Profile of the Yellow Clam Mesodesma mactroides (Mesodesmatidae) from the Southern Coast of Rio Grande do Sul,Brazil

The yellow clam Mesodesma mactroides (Mesodesmatidae) is a sandy beach bivalve that is distributed from Rio de Janeiro, Brazil, to the south of Buenos Aires province, Argentina. The yellow clam population has been declining in recent decades. To increase our understanding of this species, we evaluated the immunological status of yellow clams collected during different seasons from various areas in the state of Rio Grande do Sul, Brazil. We characterized the hemocytes, determined the differential hemocyte counts (DHCs), calculated the apoptotic index, and evaluated the incidence of parasites in yellow clams through histological analysis. We identified two types of hemocyte (hyaline and granular) that showed significant variation in DHCs among sampling areas during the summer and winter. The apoptotic index only exhibited significant variation during the summer. Histopathological analysis results did not significantly differ among sampling areas. This work demonstrated that environmental variation (e.g., temperature and salinity) associated with anthropogenic actions may be affecting the immune system of yellow clams. However, more studies are needed to determine the full influence of these factors on the yellow clam’s immune system and thus contribute to future management and aquaculture of the species.

Received May 10, 2015; accepted October 28, 2015.     

IMAGING DIAGNOSIS — USE OF MULTIPHASE COMPUTED TOMOGRAPHIC UROGRAPHY IN THE DIAGNOSIS OF URETERAL TEAR IN A 6‐DAY‐OLD FOAL

A 6‐day‐old foal was evaluated for depression and inappetence. After initial stabilization, the filly developed a hypochloremic metabolic alkalosis and persistent hypercreatinemia. Abdominal ultrasound revealed fluid accumulation around the left kidney and a well‐defined rounded fluid filled structure dorsal to the urinary bladder. Computed tomography revealed a partial tear of the left ureter with distension of the retroperitoneal membrane. Exploratory celiotomy was performed to allow left kidney nephrectomy. At 6 months follow‐up, the filly was growing normally without complications. Ultrasound and CT imaging in this case provided an accurate diagnosis and a presurgical aid to select the appropriate therapeutic approach.