Neoplastic pleural effusion and intrathoracic metastasis of a scapular osteosarcoma in a dog: a multidisciplinary integrated diagnostic approach

A 10‐year‐old, female spayed mixed‐breed or cross‐bred dog was referred to the Small Animal Teaching Hospital of the University of Liverpool due to tachypnea, dyspnea, and pleural effusion not responding to diuretics and antibiotics. The chest was drained and cytology of the pleural fluid was consistent with a modified transudate with presence of atypical cells initially attributed to mesothelial hyperplasia and dysplasia. Computed tomography detected, in addition to the bilateral pleural effusion, diffuse pleural thickening, multiple pleural and pulmonary nodules, and a mineralized and lytic mass in the left scapula. Imaging findings were suggestive of a primary bone tumor with intrathoracic metastasis. Cytology of the left scapular and pleural masses revealed a malignant neoplasm highly suggestive of osteosarcoma. The diagnosis was confirmed by demonstration of a positive cytochemical reaction for alkaline phosphatase on prestained cytology slides. This finding prompted review of the initial interpretation of the pleural effusion cytology. The presence of neoplastic osteoblasts in the thoracic fluid was identified by a combination of cytochemistry, cell pellet immunohistochemistry, and transmission electron microscopy findings. In this report, a multidisciplinary integrated diagnostic approach was used to diagnose and confirm a neoplastic pleural effusion due to osteosarcoma metastasis in a dog.     

Sediment transport below a small alpine reservoir desilted by controlled flushing: field assessment and one-dimensional numerical simulation

Purpose

Sediment transport and riverbed sedimentation were investigated in an alpine stream below a small hydropower reservoir desilted by a controlled sediment flushing (CSF) operation. The term “controlled” refers to the operational tasks implemented to mitigate the downstream environmental impact of the operation. The experimental dataset acquired before, during, and after the CSF was also used to carry out and calibrate a one-dimensional sediment transport model of the monitored event.

Materials and methods

The investigated reservoir is located in the central Italian Alps, and its original storage was 160,000 m³, about 30% filled by a mixture of sand and silt/clay before the CSF. Downstream sediment concentration was controlled by releasing clear water from upstream reservoirs and regulating the work of earth-moving equipment in the emptied reservoir. A 3.6-km-long reach with average slope of 0.015 was monitored: concentration and grain size of suspended sediment were measured during the CSF and the riverbed alteration was evaluated by volumetric sampling and measurements of the deposits’ thickness. Sedimentation and River Hydraulics—One Dimensional (SRH-1D) was used to simulate sediment transport during the monitored CSF. Model parameters were calibrated by comparing the computed and the observed amount of sediment deposited along the study reach.

Results and discussion

Sediment flushing was carried out in October 2010 for 3 days. Ca. 16,000 m³ of sediment were evacuated, representing approximately 30% silt/clay and 70% sand. 2.4 Mm³ of clear water was released to reduce sediment concentration and increase transport capacity downstream. About 3000 m³ of sand was deposited in the study reach after the CSF, with maximum height up to 0.2 m. Although the riverbed before the CSF was simply set as mono-granular, after calibrating the parameters, good agreement was achieved between the depositional pattern computed by SRH-1D and the one observed, both in terms of deposit thickness and grain size of deposited sediment. The sensitivity analysis revealed a major role of the parameters controlling bed mixing processes in affecting the simulated deposition after the CSF.

Conclusions

Sediment below 0.1 mm in diameter was not detected in river deposits after the flushing: the effects on river biota associated with substrate clogging by very fine sediment were therefore minimized. After proper calibration, 1-D sediment transport modeling can effectively support the planning of CSF operations: to minimize the downstream environmental effects, concurrently achieving acceptable flushing efficiency, the analyzed scenarios as well as the model outputs need to be carefully evaluated from a multidisciplinary perspective.

     

Response of common wheat varieties to organic and conventional production systems across Italian locations,and implications for selection

Specific breeding for organic systems may help reduce their yield gap relative to conventional systems by exploiting genotype × system (GS) interaction. Likewise, specific breeding for distinct subregions within a region could capitalize on genotype × location (GL) interaction. Grain yield and test weight of common wheat varieties were evaluated under organic and conventional systems in ten locations spanning from northern to southern Italy, with the objectives of: (i) comparing production systems; (ii) investigating the extent of GS and GL interactions and their relationship with genotypic and environmental characteristics; and (iii) preliminarily comparing, in terms of predicted selection gains, different strategies to cope with GS and GL effects. These effects were investigated in the 2-year Data set 1 including seven genotypes. GS effects were also assessed in the annual Data sets 2 and 3 including 13 and 11 genotypes, respectively. The yield reduction of organic systems relative to conventional ones averaged 28% in Data set 1, 29% in Data set 2 and 14% in Data set 3. Organic systems also tended to a modest test weight reduction. Genetic correlations between systems ranged from high to very high (0.88 ≤ r_g ≤ 0.98) for yield and test weight, owing to nil or limited GS interaction. Broad-sense heritability tended to be higher in conventional systems than organic ones for yield in two data sets (mainly due to lower experimental error) while being similar in the two systems in the other cases. Predicted selection gains suggested nil (yield) or very modest (test weight) advantage of direct selection in organic systems relative to indirect selection in conventional systems, when targeting organic systems. The scope for selection only in conventional systems was reinforced when comparing predicted gains for selection scenarios which target both systems in relation to their foreseeable marketing importance. GL effects for yield and test weight were significant and were modeled by additive main effects and multiplicative interaction analysis. Site classification based on GL effects for yield revealed a larger subregion A including northern and central Italy and a smaller subregion B comprising southern Italy, accordingly with previous, independent studies. Yield selection only in subregion A (with indirect selection gain for subregion B) implied slightly higher predicted gain for A (+4%) and much lower gain for B (−24%) relative to independent, direct selection in each subregion. Selection for specific geoclimatic subregions may have greater importance than selection for specific production systems.     

Assessment of synthetic chemicals for disruption of Rhynchophorus ferrugineus response to attractant-baited traps in an urban environment

The red palm weevil (RPW), Rhynchophorus ferrugineus Olivier (Coleoptera, Curculionidae), is one of the most severe pests of ornamental palm species in urban areas of Mediterranean countries. Aiming to discover inhibitory semiochemicals for RPW population management in urban environments, we conducted electroantennographic (EAG) screenings of 17 commercially available synthetic compounds, representing three groups of plant volatiles (isoprenoids, phenyl propanoid derivatives and fatty acid derivatives) known for their repellent effects toward insects. These tests were followed by trap-based screenings of EAG-active menthone, α-pinene and methyl salicylate, singly and in combination, under urban conditions. In EAG bioassays, RPW antennae of both sexes showed positive dose-dependent responses to 13 of the 17 synthetic chemicals with significant differences among them. In field trapping experiments, conducted in the city of Palermo, Italy, from weeks 31 to 38 in 2010 and 2011, α-pinene, tested singly or in combination with methyl salicylate (2010) or menthone (2011), reduced trap catches by about 30% to 40%. Methyl salicylate and menthone alone were inactive. In conclusion, RPW is significantly affected by α-pinene. This isoprenoid is a promising disruptant for semiochemical-based management of this pest. In addition, identification of a large number of EAG-active chemicals could serve as a database for future design of active repellents or attractants of RPW adults.     

Lucerne shoot and root traits associated with adaptation to favourable or drought-stress environments and to contrasting soil types

Cultivar × location interaction for lucerne forage yield across northern Italy is large, repeatable and associated with summer drought-stress level and soil type. The objectives of this study were: (i) to investigate the genotypic factors associated with cultivar adaptive responses to drought-stress and soil factors; (ii) to identify adaptive traits exploitable for selection of widely adapted or specifically adapted material. Aerial dry matter (DM) over 12 harvests and shoot traits of 13 landraces and four varieties were evaluated in four artificial environments created by the factorial combination of drought-stress level (almost nil or high) and soil type (sandy-loam or silty-clay) (Exp. 1). Aerial and root DM over four or five harvests were evaluated in metal containers 55 cm × 12 cm × 75 cm deep for the factorial combinations of three varieties by two drought-stress levels by two soil types (Exp. 2), or six landraces by two drought-stress levels (Exp. 3). Cultivar × environment interaction was detected for forage yield, plant mortality after the second summer, leaflet size and stem weight. The environments of Exp. 1 reproduced well the variety adaptive responses across agricultural environments. The relationship of cultivar forage yield with shoot traits was environment-specific, i.e.: (i) strictly negative with plant mortality, in no-stress environments (where mortality and plant competition were severe); (ii) positive with stem number per plant and autumn-winter growth, in stress environments; (iii) positive with stem dry weight, in ‘stress/sandy-loam soil’; and (iv) positive with leaflet size, in ‘no-stress/sandy-loam soil’. Cultivars specifically adapted to no-stress or sandy-loam conditions showed consistently greater root DM across three soil layers than material with opposite adaptive response. Entry yields tended to inverse genetic correlation between two environments which represented contrasting geographical subregions and were characterized by different combinations of traits associated with higher yield, i.e.: (i) higher root biomass, lower plant mortality and larger leaflets, for ‘no-stress/sandy-loam soil’; and (ii) more stems per plant and greater autumn-winter growth, for ‘stress/silty-clay soil’. The difficulty of yield-based selection for wide adaptation may hardly be overcome by selection based on adaptive traits. Adaptive trait-based selection for specific adaptation may be envisaged especially for ‘stress/silty-clay soil’ conditions, for which the relevant traits are inexpensive to record, not correlated, and not subject to cultivar × environment interaction.     

Digestive enzymes of the crustaceans Munida and their application in cheese manufacturing: a review

Crustaceans Munida (fam. Galatheideae, ord. Decapodi) were fished in the Southern Adriatic Sea and their proteolytic activities were characterized and tested for potential application in cheese manufacturing. Enzymes extracted from whole crustaceans, mainly serine proteases, showed high caseinolytic and moderate clotting activities. Analysis by 2D zymography of the digestive enzymes extracted from Munida hepatopancreas, showed the presence of several isotrypsin- and isochymotrypsin-like enzymes in the range of 20-34 kDa and 4.1-5.8 pI. Moreover, specific enzymatic assays showed the presence of aminopeptidases and carboxypeptidases A and B. Overall, optimum activity was achieved at pH 7.5 and 40-45 °C. Caseinolytic activity, determined both spectrophotometrically and by SDS gel electrophoresis, indicated higher activity on β-casein than on α-casein. Miniature cheddar-type cheeses and Pecorino-type cheeses were manufactured by adding starter, rennet and Munida extracts to milk. Reverse-phase HPLC and MALDI-ToF mass spectrometry showed a more complex pattern of proteolytic products in cheeses made using Munida instead of chymosin. Munida extracts were found to degrade the chymosin-derived β-casein fragment f193-209, one of the peptides associated with bitterness in cheese. In conclusion, Munida digestive enzymes represent a promising tool for development of new cheese products and shorten cheese ripening when used either alone or in addition to calf rennet.     

Cytokine expression, glucocorticoid and growth hormone changes after porcine reproductive and respiratory syndrome virus (PRRSV-1) infection in vaccinated and unvaccinated naturally exposed pigs

The objective of this paper was to study the changes of some cytokines and neuroendocrine hormones in vaccinated and unvaccinated pigs that were naturally infected by a PRRSV-1 (porcine reproductive and respiratory syndrome virus) heterologous field strain. We analyzed gene expression of pro-inflammatory (TNF-α, IL-1β, MCP-1, IL-6), pro-immune (IFN-γ) and anti-inflammatory cytokines (IL-10) in PBMC, as well as hormonal (GH and cortisol) levels in blood samples of pigs obtained in a field trial previously reported [Martelli P, Gozio S, Ferrari L, Rosina S, De Angelis E, Quintavalla C, et al. Efficacy of a modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine in pigs naturally exposed to a heterologous European (Italian cluster) field strain: clinical protection and cell-mediated immunity. Vaccine 2009;27:3788-99]. All vaccinated pigs showed an increase in pro-inflammatory and pro-immune cytokine gene expression with respect to controls and a prompt increase in GH that could be consistently associated with pro-inflammatory cytokines in sustaining innate immunity; moreover, the higher levels of cortisol indicates the activation of the hypothalamus-pituitary-adrenal (HPA) axis response. In contrast, unvaccinated pigs showed down-regulation of the cortisol and GH responses, and the pro-inflammatory and pro-immune cytokines remained at a basal or low level, with an increase of TNF-α and IL-6 in association with a higher level of IL-10 in the late phase of natural infection. The associated trends of pro-inflammatory and anti-inflammatory cytokines together with the cortisol level demonstrate that a previous vaccination promotes an early immune responsiveness in pigs and a more efficient control of inflammation in the late phase of infection with a heterologous PRRSV isolate; both events could sustain clinical protection.     

One dose of a porcine circovirus 2 subunit vaccine induces humoral and cell-mediated immunity and protects against porcine circovirus-associated disease under field conditions

This study investigated the efficacy of a one-dose porcine circovirus 2 (PCV2) subunit vaccine based on the PCV2 Cap protein expressed in a baculovirus system on two different farms at which a history of porcine circovirus-associated disease (PCVD) was present. Morbidity, mortality, average daily weight gain, carcass weight, PCV2 load in serum and vaccine immunogenicity were assessed. Serology to porcine reproductive and respiratory syndrome virus (PRRSV) and Mycoplasma hyopneumoniae was performed. A double-blind, randomised, and controlled field trial was performed distributing 818 piglets between two treatment groups. At inclusion (weaning at 21 ± 3 days of age), 408 animals (group B) received a 2-mL intramuscular dose of Porcilis PCV(?) (vaccinated group). Controls (group A, 410 pigs) received 2 mL of the adjuvant Diluvac Forte(?) intramuscularly. Weights were recorded at inclusion and at 12 and 26 weeks of age, and the average daily weight gain (ADWG) was calculated. The carcass weights of the pigs from farm 2 were recorded at slaughter (274 days old). All dead animals (died or culled) underwent autopsy to classify them as PMWS-affected or not. At each farm, blood samples were taken from 22 pigs/group for serologic studies. A beneficial effect was found after vaccination with a single dose of a PCV2 Cap vaccine against PCVD. The vaccination reduced the mortality rate and morbidity, reduced PCV2 viremia and viral load, improved productive performances (e.g. ADWG: +70 g/day between 12 and 26 weeks of age when viremia and the specific disease occurred) as well as carcass weight at slaughter age (+4.5 kg). These effects were associated with virologic and clinical protection from the immunogenicity of the vaccine measured as activation of both a humoral and a cellular immune response.     

Intra-varietal genetic diversity of the grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) cultivar ‘Nero d’Avola’ as revealed by microsatellite markers

The Sicilian grape cultivar ‘Nero d’Avola’ is among the oldest and most cultivated in the island, taking part in the production of several red wines exported worldwide, including DOC wines (Etna Rosso and Cerasuolo di Vittoria). Due to the ancient origin and repeated clonally propagation of the cultivar, phenotypic variability has been observed. Clone identification in this important cultivar has so far relied on phenotypic and chemical traits analyses, often affected by environmental conditions. Genetic markers, such as microsatellites, are particularly useful for cultivar identification, parentage testing, pedigree reconstruction and population structure studies. In the present paper, microsatellites were used to analyze the intra-varietal genetic diversity among 118 plants of ‘Nero d’Avola’, collected in 30 vineyards displaced in different areas of Sicily. Out of 22 microsatellites, 11 showed polymorphism among samples and 15 different phylogenetic groups were identified. Results show that ‘Nero d’Avola’ actually comprises different genetic profiles, although most of clones share a common origin.