Genetic diversity via REML-BLUP of ex situ conserved macauba [Acrocomia aculeata (Jacq.) Lodd. ex Mart.] ecotypes

Genetic Resources and Crop Evolution - Macauba [Acrocomia aculeata (Jacq.) Lodd. ex Mart.], a palm tree native to Brazil, has a high potential as an alternative source for vegetal oil production....     

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.     

Initial cooling time before freezing affects post‐thaw quality and reproductive performance of rabbit semen

The aim of this study was to investigate the effect of initial cooling time at 5°C during semen cryopreservation on post‐thaw quality and reproductive performance of rabbit semen. Pooled semen samples (n = 6) were divided into two subsamples and cooled at 5°C for 45 or 90 min. After cooling, the semen samples were diluted to a ratio of 1:1 (v:v) with a freezing extender composed of Tris‐citrate‐glucose (TCG) containing 16% of dimethylsulfoxide and 0.1 mol/L sucrose. The semen was subsequently loaded in 0.25 ml straws, equilibrated at 5°C and frozen in liquid nitrogen vapor. After thawing, sperm motility, viability, osmotic resistance, acrosome and DNA integrity were assessed. Our results indicate that the longer cooling time, that is, 90 min before cryopreservation significantly improves sperm post‐thaw viability, motility and fertility. In fact, reproductive performances obtained with semen frozen after a 90 min cooling time were similar to those produced by fresh semen insemination. Hence, the present research provides an effective freezing protocol for rabbit semen that will allow for the creation of a sperm cryobank for the conservation of Italian rabbit genetic resources, as well as the use of frozen semen doses in commercial farms.     

Evaluation of EDTA hematology tubes for collection of blood samples for tests of secondary hemostasis in dogs

OBJECTIVE: To evaluate the use of EDTA tubes for collection of blood samples for assays of secondary hemostasis in dogs. ANIMALS: 108 dogs of various ages, breeds, and sexes (19 healthy and 89 with abnormalities of secondary hemostasis). PROCEDURES: Blood samples were collected via cephalic venipuncture and transferred to sodium citrate tubes and EDTA tubes. Plasma was harvested from each type of tube for assays of concentrations of fibrinogen and D-dimer as well as prothrombin time, activated partial thromboplastin time, and antithrombin activity. Intra-assay and interassay precision and correlation coefficients for all hemostatic tests were calculated for each type of plasma sample. The effect of storage conditions on assay results for the 2 types of plasma samples was also evaluated. RESULTS: Results of hemostatic tests were highly correlated between citrated and EDTA-treated plasma samples. Intra-assay imprecision for all hemostatic tests with the exception of D-dimer concentration was < 10% for both citrated and EDTA-treated plasma samples; interassay imprecision was higher for EDTA-treated versus citrated plasma samples. Storage of plasma samples for 1 hour did not result in significantly different assay results for either type of plasma sample, but storage for 2 hours significantly affected values for EDTA-treated plasma samples. CONCLUSIONS AND CLINICAL RELEVANCE: Although evaluation of the sensitivity and specificity of hemostatic tests that use EDTA-treated plasma samples is required, EDTA may be a suitable alternative to sodium citrate as an anticoagulant for use in hemostatic testing in conditions in which tests could be performed within 1 hour after sample collection.     

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.     

Homocysteine measurement by an enzymatic method and potential role of homocysteine as a biomarker in dogs

In humans, homocysteine (Hcy) is employed to monitor renal, cardiovascular, and other diseases and their complications. The aim of the current study was to define the analytical performances of an enzymatic method not yet validated in dogs for measuring homocysteine, and to assess the possible clinical usefulness of Hcy measurement. Using conventional approaches, the analytical performances were investigated by assessing, imprecision, inaccuracy, and interference of hemoglobin, triglycerides, and bilirubin. The possible clinical usefulness of Hcy determination was assessed by comparing the results of healthy dogs (n = 8) with those of dogs with heart disease (n = 10), inflammation (n = 6), gastrointestinal disorders (n = 7), neoplasia (n = 8), renal failure (n = 4), trauma (n = 7), and other miscellaneous diseases (n = 6). Preliminary evaluation of this enzymatic method showed it to be precise at Hcy levels close to or higher than the values in dogs with renal or cardiac disorders that had the highest Hcy levels. By contrast, at low Hcy levels, which were recorded basically in control dogs, the method suffers from high imprecision. The sample of choice is serum. The use of icteric samples should be avoided, while hemoglobin and lipids have only a minor effect on Hcy measurement. In conclusion, the enzymatic method employed in the current study provides useful information in dogs and could be used to monitor cardiac and renal disorders, in which Hcy concentrations are elevated.     

Effects of propofol on ultrasonic indicators of haemodynamic function in rabbits

OBJECTIVE: To evaluate the cardiovascular effects of intravenous propofol in rabbits. STUDY DESIGN: Randomized, prospective, experimental study. ANIMALS: Thirty-one female New Zealand White rabbits. METHODS: Rabbits were allocated to one of two groups [propofol (P) or conscious (C)]. In C (n = 16) vascular dimensions were measured using ultrasound of the left common carotid artery (ACC) and the abdominal aorta (AA). Group P (n = 15) received propofol 4.0-8.0 mg kg(-1) intravenously (IV). Anaesthesia was maintained with propofol at 1.2-1.3 mg kg(-1) minute(-1). Subsequently, three propofol injections (8 mg kg(-1)) were given. Before and for 10 minutes after each injection the following vascular and haemodynamic variables were recorded (a) at the ACC after the first injection; and (b) at the AA after the second injection: vessel diameter [D, (mm)], peak systolic, minimum diastolic, end-diastolic and average blood flow velocities [psBFV, mdBFV, edBFV, Vave (cm second(-1))], average volumetric flow [VFave (mL s(-1))], resistance index (RI) and pulsatility index (PI) mean arterial pressure (MAP), heart rate (HR), arterial oxygen saturation (SpO(2)) and end-tidal CO(2) (Pe'CO(2)). Echocardiography was performed after the third propofol bolus injection to investigate changes in cardiac parameters [fractional shortening, FS (%)]. RESULTS: Intravenous propofol injections caused a significant decrease in vessel diameter, volumetric flow and edBFV, and significant increases in psBFV, RI and PI. Baseline levels for vessel diameter and psBFV were restored 6-8 minutes after injection. Propofol injection decreased FS significantly by 7 minutes after injection while MAP and HR were significantly reduced for 4 minutes. CONCLUSION AND CLINICAL RELEVANCE: Injections of propofol (8 mg kg(-1)) produced an immediate, transient decrease in vascular diameters, a significant decrease in ventricular performance and an increase in peripheral vascular resistance (ACC and AA). Propofol should probably not be or only carefully used in rabbits with ventricular dysfunction.     

Partially defatted black soldier fly larva meal inclusion in piglet diets: effects on the growth performance,nutrient digestibility,blood profile,gut morphology and histological features

Background: The aim of this trial was to investigate the effects of different inclusion levels of a partial y defatted black soldier fly(BSF, Hermetia il ucens L.) larva meal on the growth performance, nutrient digestibility, blood profile, gut morphology and histological features of piglets. A total of 48 newly weaned piglets were individual y weighed(initial body weight(IBW): 6.1 ± 0.16 kg) and randomly al ocated to 3 dietary treatments(4 boxes as replicates/treatment and 4 animals/box). BSF larva meal was included at increasing levels(0% [BSF0], 5% [BSF5] and 10% [BSF10]) in isonitrogenous and isoenergetic diets formulated for two feeding phases: I(from d 1 to d 23) and I(from d 24 to d 61). The weight gain(WG), average daily gain(ADG), average daily feed intake(ADFI) and feed conversion ratio(FCR) were calculated for each feeding phase and for the whole trial. The haematochemical parameters and nutrient digestibility of the piglets were also evaluated. A total of 3 piglets per box were slaughtered on d 61 and the slaughtered piglets were submitted to morphometric investigations and histopathological examinations.Results: No overal significant differences were observed for growth performance(P 0.05), except for the ADFI of phase I, which showed a linear response to increasing BSF meal levels(P 0.05, maximum for the BSF10 group). Dietary BSF meal inclusion did not significantly influence the blood profile, except as far as monocytes and neutrophils are concerned,and these showed a linear and quadratic response, respectively, to increasing BSF meal levels(P 0.05, maximum for the BSF10 and BSF5 groups, respectively). On the other hand, the nutrient digestibility, gut morphology and histological features were not affected by dietary BSF meal inclusion(P 0.05).Conclusions: The obtained results show that a partial y defatted BSF larva meal can be used as a feed ingredient in diets for weaned piglets without negatively affecting their growth performance, nutrient digestibility, blood profile, gut morphology or histological features.