Effects of Addition of Tissue‐Type Plasminogen Activator in In Vitro Fertilization Medium on Bovine Embryo Development and Quality

Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue‐type plasminogen activator (t‐PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus‐oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t‐PA and/or its inhibitor epsilon‐aminocaproic acid (control, t‐PA, t‐PA+ε‐ACA, ε‐ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t‐PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε‐ACA. PAA in the treated group was significantly reduced by ε‐ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t‐PA‐modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t‐PA. In conclusion, it appears that excessive t‐PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.     

Large‐scale fuzzy rule‐based prediction for suitable chestnut ink disease sites: a case study in north‐east Italy

In the past few decades, economic interest in the cultivation of chestnuts for both timber and nut production has resurfaced in the Mediterranean area. However, chestnut cultivation has suffered in recent years from the spread of exotic pests, such as the gall wasp Dryocosmus kuriphilus, and from the resurgence of previously present diseases, most likely due to anomalous climate dynamics. This is the case with chestnut ink disease, caused by the soilborne pathogens Phytophthora cinnamomi and P. cambivora. Scientific and technical support in monitoring and management, that utilizes new forecasting approaches incorporating related environmental variables, is therefore essential. The main aim of this study was to develop a mathematical model assessing the potential for the establishment of chestnut ink disease at a large scale. Towards this goal, fuzzy rule‐based theory was applied to the environmental variables associated with host presence, pathogens' ecological niches and ink disease symptoms expression. The effectiveness of the rule‐based modelling outcomes, provided with uncertainty maps to facilitate their correct interpretation, was confirmed by detailed field data collected from a large chestnut‐growing area where ink disease has been increasing in recent years. The final model gave consistent predictions for disease presence. For this reason, it represents a flexible and valuable decision‐support tool to forecast which sites are at risk from CID.     

Deleterious Effects of Progestagen Treatment in VEGF Expression in Corpora Lutea of Pregnant Ewes

The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri‐implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen‐treated group showed smaller corpora lutea than cloprostenol‐treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen‐treated group, which even showed a decrease between days 11 and 13. In progestagen‐treated sheep, VEGF expression in granulosa‐derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.     

Antineurotoxic activity of Galactia glaucescens against Crotalus durissus terrificus venom

Ethanolic extract of leaves of Galactia glauscescens (GGE) at concentration of 100 and 500 mug/ml prevented the neuromuscular paralysis induced by Crotalus durissus terrificus venom on mouse phrenic nerve-diaphragm preparation.     

Survey onYersinia Enterocolitica and related bacteria in shellfish

Results from examining shellfish collected from ‘safe’ (2–34Escherichia coli/100 mL) and ‘unsafe’ (> 34E. coli/100 mL) waters for the presence ofYersinia enterocolitica and related bacteria are reported. The recovery of these microorganisms was low in bivalve mollusks destined for food consumption, high in those derived from prohibited areas. No human pathogenic bio-serogroup f Yersinia was found during the entire survey.     

Insecticidal efficacy of beauveria bassiana,diatomaceous earth and fenitrothion against rhyzopertha dominica and tribolium castaneum on stored wheat

Bioassays were conducted in stored grains to evaluate the combined use of the entomopathogenic fungus Beauveria bassiana, diatomaceous earth (DE) and fenitrothion against adults of Rhyzopertha dominica and Tribolium castaneum. The insecticide agents were applied to wheat as follows: the fungus at 1 × 10⁸ conidia/kg of wheat, DE at 3 g/kg of wheat and fenitrothion at a rate of 0.15 ml/kg of wheat (25% of field dose rate). Surviving insects and progeny production were monitored at thirty-day intervals until four months. For both insect species, significantly less progeny was produced on wheat treated with fenitrothion and DE + B. bassiana formulations in comparison with controls. The effect of insecticides on the grain damage, germination power of wheat and bulk density was also evaluated. After four months, B. bassiana mixed with DE reduced the damaged insect grains by 50% in comparison with both fenitrothion and control treatments. Trials using DE caused a reduction in bulk density while there were no significant differences for germination testing between treatments.     

Fundamental evaluation of the impact of high Hydrostatic Pressure on oat batters

The use of grains alternative to wheat or rye is a challenging task for cereal technologists, and currently new technologies are under investigation as tools to improve the performances of these alternative grains. In this work the effects of high Hydrostatic Pressure (HP) on oat batters were investigated. Oat batters were treated for 10 min at 200, 300, 350, 400 or 500 MPa. Scanning electron microscopy and bright field microscopy showed that high HP significantly affected oat batter microstructure, and both starch and proteins were affected. Treatment at high HP significantly improved batter viscosity and elasticity. At pressures ≤300 MPa the increase in the viscous component was higher than the increase in the elastic component. On the contrary, at pressures ≥350 MPa the elastic component was predominant. Differential scanning calorimetry revealed that high HP induced starch gelatinisation, which started at 300 MPa and was almost complete after treatment at 500 MPa. High HP also affected water- and salt-soluble as well as urea-soluble oat proteins. Analysis of proteins soluble in different buffers revealed that pressures ≥300 MPa induced the formation of urea-insoluble complexes and/or disulfide bonds. Overall, the extent of starch gelatinisation and protein modification was dependent on the applied pressure, but the results collected so far clearly show that high HP can be used to improve the functionality of oat batters.