Search

Octopus vulgaris is a commercially valuable species. It is overexploited in the natural environment and is considered to be an innovative species for aquaculture. However, large-scale farming is generally designed only based on economic requirements, disregarding any form of enrichment that induces the natural behavior of aquatic species. Although many studies have shown the influence of environmental enrichment on terrestrial vertebrates, fish, and cephalopod mollusks, information on the effect of environmental enrichment on the body patterns of O. vulgaris is limited. Therefore, in this study, we assessed how different environmental conditions (Basic vs. Enriched) affect sub-adults of O. vulgaris kept in recirculation systems, through qualitative–quantitative studies of the main body patterns and their potential application in the commercial production of this species. The results indicated that octopuses kept in the enriched environment showed several body patterns and gained a significantly higher weight than those kept in the basic environment. The body patterns displayed by the individuals kept in the basic environment were similar to those exhibited under situations of hostility and inter/intra-specific conflict. Hence, the environment of octopuses needs to be enriched, especially for the large-scale production of this species.

Introduction. We have recently developed a new miniaturized device for extracorporeal ultrafiltration to be used in patients with fluid overload: Artificial Diuresis-1, or AD1 (Medica S.p.A., Medolla, Italy). The device has a reduced priming volume and operates at very low pressure and flow regimes and is designed to perform extracorporeal UF at bedside. After accurate experiments carried out in vitro, we report in this paper the results of in vivo tests ultrafiltration session carried out in selected animals according to veterinary best practice. Materials and methods. The AD1 kit is pre-filled with sterile isotonic solution and operates with a polysulfone mini-filter MediSulfone (Polysulfone at 50000 Dalton). A collection bag with a volumetric scale is connected to the UF line and the ultrafiltrate is obtained by gravity based on the height at which the ultrafiltrate collection bag is placed. Animals were prepared and anesthetized. Jugular vein was cannulated with a double lumen catheter. Three six hours sessions of ultrafiltration were scheduled with a target fluid removal of 1500 ml. Heparin was used as anticoagulant. Results. In all treatments the target value of ultrafiltration was obtained in the absence of major clinical or technical problems with a maximum deviation from the scheduled ultrafiltration rate lower than 10%. The device resulted safe, reliable, accurate and easily usable thanks to a user friendly interface and the very small dimensions. Conclusions. This study opens the way for clinical trials in different settings including departments with low intensity of care and even in ambulatory centers or patient’s home.

Purpose AL2106 is a new medical device based on a mixture of chondroitin sulphate in a xyloglucan and glycerol solution made to maximize its bioadhesive capability to the esophageal mucosa. The aim of the present study was twofold to evaluate the AL2106 protective effect on the esophageal mucosa when exposed to an acidic solution mimicking gastric reflux and to assess the resilience of this effect to saline washing. Materials and Methods A porcine ex vivo model was used and the effects of the new medical device were compared to a sodium alginate suspension (SAS) already present on the market which was assumed as reference. Mucosal damage was induced in 19 porcine esophagi by perfusion with an acidic solution added with pepsin, and Evans blue dye (EBD) tissue uptake was used as an indicator of mucosal permeability. The EBD penetration, expressed as EBD µg/g of dry tissue, was assessed in specimens of untreated damaged mucosa and in specimens treated with AL2106 or SAS. The same evaluation was carried out after washing with normal saline. Results Both topical agents tested significantly reduced the EBD uptake by more than 60% (AL2106 8.4±4.5, SAS 3.6±2.7 vs control 23.2±13.1, p

Abstract The first weeks of life represent a crucial stage for microbial colonization of the piglets’ gastrointestinal tract. Newborns’ microbiota is unstable and easily subject to changes under stimuli or insults. Nonetheless, the administration of antibiotics to the sow is still considered as common practice in intensive farming for pathological conditions in the postpartum. Therefore, transfer of antibiotic residues through milk may occurs, affecting the piglets’ colon microbiota. In this study, we aimed to extend the knowledge on antibiotic transfer through milk, employing an in vitro dedicated piglet colon model (MICODE—Multi Unit In vitro Colon Model). The authors’ focus was set on the shifts of the piglets’ microbiota composition microbiomics (16S r-DNA MiSeq and qPCR—quantitative polymerase chain reaction) and on the production of microbial metabolites (SPME GC/MS—solid phase micro-extraction gas chromatography/mass spectrometry) in response to milk with different concentrations of amoxicillin. The results showed an effective influence of amoxicillin in piglets’ microbiota and metabolites production; however, without altering the overall biodiversity. The scenario is that of a limitation of pathogens and opportunistic taxa, e.g., Staphylococcaceae and Enterobacteriaceae, but also a limitation of commensal dominant Lactobacillaceae, a reduction in commensal Ruminococcaceae and a depletion in beneficial Bifidobactericeae. Lastly, an incremental growth of resistant species, such as Enterococcaceae or Clostridiaceae, was observed. To the authors’ knowledge, this study is the first evaluating the impact of antibiotic residues towards the piglets’ colon microbiota in an in vitro model, opening the way to include such approach in a pipeline of experiments where a reduced number of animals for testing is employed. Key points • Piglet colon model to study antibiotic transfer through milk. • MICODE resulted a robust and versatile in vitro gut model. • Towards the “3Rs” Principles to replace, reduce and refine the use of animals used for scientific purposes (Directive 2010/63/UE). Graphical abstract

In the development of an aero-engine combustor, the definition of a preliminary design is a practice in which know-how, product experience and design rules are focal in deriving a configuration able to meet the functional requirements. Several configurations, and hence multiple geometries resulting in different behaviours, are iteratively analysed in this phase to extensively explore the design space. In this context, an automated procedure ranging from preliminary design to life estimation is necessary and crucial. A framework in which the tools employed in the design workflow are integrated and the low-added-value tasks are automated can allow the reduction of time per analysis within the loop and the enhancement of the procedure’s robustness. In this paper will be presented the Combustor Design System Integration (DSI), a methodology aimed at easing and streamlining the design process of aero-engine combustors. To do this, digitization has been taken as the common thread for developing a data-centric approach. The logic behind the procedure will be reported, to focus then on the aero-thermal preliminary design. The procedure, for this phase, is composed of three main integrated components: a CAD generation system, which collects all the geometries for creating an exportable 3D model, a 1D thermal solver for the positioning and sizing of the aero feature on liners (i.e. cooling, dilution...) and a CFD environment with automated pre/post processing operations for reacting-flow analysis. The aim of this work is to contextualize the DSI approach in the combustor design process and to provide a first description of the methodology designed and developed in GE Avio. For that purpose, a straight-through configuration — the lean combustor NEWAC developed in the homonymous EU project — will be exploited as a test case. The development of the procedure is still in progress, so a validation through test cell data comparison, as well as highly-resolved CFD results, will be the subject for future papers.

As far as the preliminary thermal design of gas turbine components is concerned, 1-D codes are still widely used in standard industrial practice. Among the different components, the combustor is one of the most critical ones and its thermal design still greatly affects the reliability and life of the entire engine. During the initial phases of the design process, parameters are often roughly known. For this preliminary phase, a low-order approach is preferred instead of a high-fidelity simulation: the exploration of the whole space is extremely important to better understand the behavior of the system and to focus on the design objectives. Uncertainty quantification (UQ) methods, mainly developed in recent years and applied in many fields, are useful tools for the preliminary design phase and provide support during the whole design process. The objective of this work is to estimate the main sources of uncertainties in the design phase of an aeroengine effusion cooled combustor. The test case is based on a full annular lean-burn combustor, tested during the LEMCOTEC (Low EMissions COre-engine TEChnologies) European project. Among the test points investigated in the experimental campaign, the Approach condition is here analyzed. The inner liner is taken into consideration to investigate the metal temperature. Therm-1D, a 1-D in-house simulation code, is used to model the combustor and the open-source tool DAKOTA is adopted for the uncertainty quantification analysis. The baseline case of the combustor is studied and several uncertainty analyses are investigated. They are divided into 3 main groups: geometrical, tuning modelling parameters and thermal loads. For each group, the most relevant parameters are considered as a source of input uncertainty. In particular, a classical Monte Carlo approach is compared with four innovative polynomial-chaos approaches for each group: Gauss quadrature, total order with LHS sampling, stochastic collocation, and Smolyak. The analyses proved how the last two methods give the best results with a sensible lower amount of simulation (depending on the number of input variables). Lastly, results are compared with experimental data to achieve a better understanding of the most relevant input parameters and the propagation of their uncertainty on the results.

The preliminary design of an aero-engine combustor is a multidisciplinary process that involves an extensive and systematic analysis of the design space. Simulation-driven approaches, in which several design configurations are numerically analyzed, may lead to heterogeneous models interacting with each other, sharing miscellaneous information within the process. Iterative and user-defined approaches, moreover, are inefficient when multiple and conflicting requirements are in place. To rely on integrated design methodologies has been demonstrated to be beneficial, especially if adopted in a structured approach to design optimization. In this paper, the application of the Combustor Design System Integration (DSI) to the definition of an optimal combustor preliminary configuration will be presented. Given a combustor baseline design, the multi-objective optimization problem has been defined by targeting an optimal distribution for temperature profiles and patterns at the combustor’s exit. Dilution port characteristics, such as hole number and dimension as well as the axial position of the row have been selected as design variables. To guarantee a water-tight design process while minimizing the user effort, the DSI tools were included in a dedicated framework for driving the optimization tasks. Here, a proper CFD domain for RANS, constituted by the flame tube region extended to the dilution port feeds, was adopted for imposing the air split designed for the combustor. Concerning a “complete” combustor sector, this allows a reduction in the computational effort while still being representative for its aero-thermal behavior. The optimization task was performed using a Response Surface Method (RSM), in which multiple, specific combustor configurations were simulated and the CFD result elaborated to build a meta-model of the combustor itself. Finally, the suitability of the resulting optimized configuration has been evaluated through an “a posteriori” analysis for thermal conditions and emission levels (NOx and CO). A lean combustion concept developed by Avio Aero with the aim of the homonymous EU research project, the NEWAC combustor, has been considered as test case.

Roe deer are seasonal breeders with a complete yearly testicular cycle. The peak in reproductive activity is recorded during summer, the rutting period, with the highest levels of androgens and testicular weight. Melatonin plays a pivotal role in seasonal breeders by stimulating the hypothalamus–pituitary–gonads axis and acting locally, in different species, its synthesis within testes has been reported. The aim of this study was to evaluate the physiological melatonin pattern within roe deer testes by comparing data obtained from animals sampled during pre- and post-rut periods. Melatonin was quantified in testicular parenchyma, along with the genetic expression of enzymes involved in its local synthesis (AANAT and ASMT) and function (UCP1). Melatonin receptors, MT1-2, were quantified both at protein and gene expression levels. Finally, to assess changes in reproductive hormonal profiles, testicular dehydroepiandrosterone (DHEA) was quantified and used for a correlation analysis. Melatonin and AANAT were detected in all samples, without significant differences between pre- and post-rut periods. Despite DHEA levels confirming testicular involution during the post-rut period, no correlations appeared between such involution and melatonin pathways. This study represents the first report regarding melatonin synthesis in roe deer testes, opening the way for future prospective studies in the physiology of this species.

Simple Summary Nowadays, the importance of breastfeeding has been very well recognized not only by the scientific world but also by public opinion. Such awareness has nonetheless put a lot of pressure on women under chronic pharmacological medication, or that simply need to alleviate common post-partum health issues, due to the lack of scientific data regarding the potential transfer to the offspring during lactation. In such a scenario, the ConcePTION project aims at creating a trusted ecosystem that can efficiently generate and disseminate reliable evidence-based information regarding the effects of medications used during pregnancy and breastfeeding to women and their healthcare providers. Due to the need for a reliable animal species to obtain scientific data, the present review summarizes the main features contributing to the lactation process in the most commonly used laboratory animal species. Abstract The present review aims to summarize the main features of mammary gland anatomy, and the physiology of lactation and colostrum/milk in the most commonly used animal species for regulatory toxicity. The final goal is the selection of a preferred animal species to be enrolled in studies investigating the potential transfer of drugs and exogenous molecules through milk, within the Innovative Medicines Initiative (IMI) funded project ConcePTION. Reference data regarding humans were also collected and analyzed in order to highlight critical similarities and differences with the studied species. Additional practical considerations were also taken into account, such as ethical consideration regarding the chosen species which affects the group size, financial implications and technical feasibility of lactation trials (e.g., ease of sampling, volume of sampling, husbandry requirements and scientific recognition). In conclusion, the present analysis of the literature confirms the complexity of the decisional process behind the choice of an animal model for in vivo trials. For some of the evaluated species, data were either poor or missing, highlighting the necessity to generate more physiological background studies for species that are routinely used in laboratory settings. Overall, when taking into consideration ethical factors, feasible group size, milk volume and ease of milk collection, and physiological similarities with humans, minipigs seem to represent the most appropriate choice.

In aquaculture, the many chemicals used as therapeutics, prophylactics and growth promoters, can accumulate in fish and spread in the environment through effluent water. Nowadays, the focus of zootechnical productions has shifted towards the search for possible natural alternatives, considered safer for animals and consumers, with lower environmental impact. Herbal products are made of natural compounds, but “natural” is not necessarily synonymous of harmless, thus toxicity needs to be tested for each composition at different doses. Indeed, since herbal products' action is often dose-dependent, negative effects may be prevented by studying the posology according to the species and life stage. In this study, the effects of two different doses (0.05 and 0.1 mL/L) of a natural-based complementary feed, Gill Fish®, were assessed on the early stages of life of farmed rainbow trout (Oncorhynchus mykiss). The product was added daily to tanks containing rainbow trout eggs during the pre-hatching period, from 30 to 36 days post fertilization. Embryos mortality (evaluated three times during the six days of treatment; t0-t2) and zootechnical indices such as biomass, weight, length and number of alevins (all evaluated 3 and 4 weeks after the end of treatment; t3-t4), were analysed to assess both direct and indirect toxicity. Furthermore, whole body cortisol (from embryos and alevins; t0-t4) was measured to highlight potential stressful effects of the treatment. For each parameter, time was a statistically significative factor, while no differences were noticed between the three groups within the same time point. These results demonstrate that Gill Fish® had no harmful nor stressful effects, both immediately and short term; this herb-based product could therefore be a good candidate for future studies with different doses and/or longer administration, to evaluate its possible beneficial effects as an alternative to chemicals in aquaculture.

Steroids, providing information regarding several biological patterns including stress and sexual behavior, have been investigated in different matrices in laboratory mice. Data regarding hair quantification, indicative of longer timespans when compared to blood and saliva, are lacking. The aim of the work was to analyze the hormonal hair profile of laboratory male mice and to investigate potential relationships with age and housing, as a potential tool for welfare assessment. Fifty-six adult male C57BL/6J and C57BL/6OlaHsd substrain mice were included in the study, housed in pairs or groups. Testosterone (T) and dehydroepiandrosterone (DHEA) were quantified by radioimmunoassay, corticosterone (CORT) by ELISA. Mean hormone levels were 6.42 pg/mg for T, 23.16 pg/mg for DHEA and 502.1 pg/mg for CORT. Age influenced all hormones by significantly increasing T and DHEA levels and decreasing CORT, only DHEA, significantly higher in grouped mice, was influenced by housing conditions. The influence of age indicates the need for accurate age-related reference intervals, while the higher levels of DHEA in grouped animals suggests that such housing practice may be beneficial for social interactions. In conclusion, it seems that hair hormones quantification may be a good tool for welfare assessment in laboratory mice and may help in refining husbandry.

Purpose Qualitative and quantitative analysis of vitreous humor (VH) is important to discriminate between physiological and pathological conditions and may be particularly helpful when using animal models for ophthalmologic research. The aim of the present study was to investigate the physiological qualitative/quantitative composition of the VH of the standard pig using a metabolomics approach based on 1H nuclear magnetic resonance (NMR) spectroscopy. A secondary aim was the characterization of the VH of the porcine model of photoreceptor degeneration induced with iodoacetic acid (IAA), widely used in the biomedical field, and comparison with the physiological one. Methods VH samples were collected from 30 pigs (17 in the physiological condition and 13 treated with IAA) upon vitrectomy and analyzed by means of 1H NMR spectroscopy for characterization. Results On all analyzed samples, 40 molecules could be identified and quantified, with lactate being the most abundant in both physiological and photoreceptor degeneration conditions. Upon comparison, only 17 molecules showed statistical differences: In the IAA group, glucose and glutamine increased, while lactate, 4-amynobutyrate, hypoxanthine, dicarboxylic acids (succinate and fumarate), amino acids with their derivatives (creatine, alanine, valine, lysine, leucine, glycine, taurine, and isoleucine), and choline with its precursor sn-glycero-3-phosphocholine decreased. Conclusions The results validate the metabolic impairment determined by glycolysis inhibition upon systemic IAA administration. In conclusion, this work represents the first characterization of the porcine VH metabolome in physiological conditions and provides additional information for the characterization and refinement of the IAA-induced photoreceptor degeneration model.

Aim: The aim of the study was to set up a porcine ex vivo model of acid-induced damage and to evaluate its performance by means of multichannel intraluminal impedance and pH (MII-pH) live recording, histology, and Evans blue (EB) permeability assay. Materials and Methods: Thirteen esophagi, collected at a slaughterhouse, were ablated of their sphincters, pinned upright on a support, and placed in a thermostatic hood at 37°C with two infusion tubes and an MII-pH probe inserted in the top end. Three esophagi (histology controls) were only left in the hood for 3.5 h before sampling, while the remaining organs underwent the experimental protocol including saline infusion and recovery recording, and acid solution infusion and recovery recording. Results: MII-pH analysis highlighted a significantly stronger decrease during acid infusion when compared to saline, but a better post-infusion recovery for saline solution. At the end of the protocol, MII was still statistically lower than baseline. The acid-damaged esophagi significantly absorbed more EB dye, and histology revealed strong mucosal exfoliation. Conclusion: The proposed model of esophageal acid damage seems to be repeatable, reliable, and achievable using organs collected at the slaughterhouse. MII recording proved to have good sensitivity in detecting mucosal alterations also in ex vivo trials.

Simple Summary The roe deer (Capreolus capreolus) is a wild, small ruminant common in Europe and Asia with a peculiar reproductive cycle. The adult male shows a complete suspension of spermatozoa production during the winter that start again in spring with the peak of sexual activity in summer (July–August). The physiological mechanism underlying such a cycle is yet to be clarified. The work aimed at the analysis of some molecules, like gelatinases (MMP) and vascular growth factor, physiologically involved in tissue remodeling, in roe deer samples collected before (June–July) and after (August–September) the rutting period. Samples were provided by hunters of the Bologna Apennines area (Italy) according to the regional hunting plan. The result showed a post-rut reduction of testicular weight and testosterone, indicative of testes involution, correlated with an increase in pro-MMP2, the latent gelatinase capable of sustaining spermatogenesis once activated. It can be assumed that gelatinases are involved in the testicular cycle and start accumulating after the rutting period to be then activated in preparation of the next reproductive season inducing spermatozoa development and migration. Future studies on this pathway during all seasonal testicular cycles will provide more information about the interesting reproductive physiology of roe deer. Abstract The roe deer (Capreolus capreolus) represents a spontaneous model of testicular inactivation: During winter, bucks show a suspension of spermatogenesis that starts again in spring and peaks during the breeding season (July–August). The underlying mechanisms to the regulation of the cyclic testicular changes are still not fully clear but seem to be imputable to the spermatogenic cell line since other testicular cell populations remain stable without apoptotic phenomena. The aim of the study was to investigate apoptosis, gelatinases (MMP2 and 9), their inhibiting factors (TIMP 1-2), and two isoforms of vascular endothelial growth factor (VEGF121 and 165) with its receptors (VEGFR1-2) in testes collected during pre- and post-rut periods, and to correlate them with testicular weight (TW) and testosterone (TEST). Testes from 18 adult sexually mature bucks were collected in Bologna Apennines (Italy). Samples were weighed and parenchyma collected. Radioimmunoassay, real-time PCR, and zymography were performed. The results showed a post-rut decrease in TW and TEST and an increase in proMMP2, also highlighting a correlation between the gelatinases and the testicular functionality. The VEGF pattern did not show modifications nor correlation with TW and TEST. Overall, gelatinases and their inhibitors, described herein for the first time in roe deer testes, seem to play an important role in the testicular cycle.

The red deer (Cervus elaphus L., 1758) is one of the largest deer species in the world. Females are seasonal polyestrous, with negative photoperiod: the increase of the night peak of melatonin determines the secretion of GnRH and, therefore, LH and FSH. To date there is little information regarding the hormonal control during pregnancy for this species; this could be due to the difficulty of sampling wild subjects, while farmed animals’ hormonal concentrations may not reflect the physiology of the animal in a natural state. In this study we evaluated the concentration of cortisol and progesterone, extracted from blood and hair, on 10 wild and pregnant red deer females. Belonging to the population of the Bolognese Apennines (Italy), the hinds were sampled in the January−March 2018 period, according to the regional selective hunting plan. Plasma progesterone (P4) ranged from a minimum of 1.9 to a maximum of 7.48 ng/mL; while hair P4 concentrations varied from 41.68 to 153.57 pg/mg. The plasma and hair cortisol ranges are respectively 0.4−2.97 ng/mL and 0.03−0.55 pg/mg; the only significant correlation was found between hair concentration of P4 and the date of death. The results of this preliminary study represent a small step towards a better knowledge of this species’ physiology during pregnancy.

Aim: The aim of the study was to set up a porcine ex vivo model of acid-induced damage and to evaluate its performance by means of multichannel intraluminal impedance and pH (MII-pH) live recording, histology, and Evans blue (EB) permeability assay. Materials and Methods: Thirteen esophagi, collected at a slaughterhouse, were ablated of their sphincters, pinned upright on a support, and placed in a thermostatic hood at 37°C with two infusion tubes and an MII-pH probe inserted in the top end. Three esophagi (histology controls) were only left in the hood for 3.5 h before sampling, while the remaining organs underwent the experimental protocol including saline infusion and recovery recording, and acid solution infusion and recovery recording. Results: MII-pH analysis highlighted a significantly stronger decrease during acid infusion when compared to saline, but a better post-infusion recovery for saline solution. At the end of the protocol, MII was still statistically lower than baseline. The acid-damaged esophagi significantly absorbed more EB dye, and histology revealed strong mucosal exfoliation. Conclusion: The proposed model of esophageal acid damage seems to be repeatable, reliable, and achievable using organs collected at the slaughterhouse. MII recording proved to have good sensitivity in detecting mucosal alterations also in ex vivo trials.

Swine (Sus scrofa) are often the 'gold standard' laboratory animal for ophthalmology research due to the anatomic and physiologic similarities between the porcine and human eye and retina. Despite the importance of this model, few tools for behavioral vision assessment in pigs are available. The aim of this study was to identify and validate a feasible and reproducible behavioral test to assess vision in a pig model of photoreceptor degeneration. In addition, a robust behavioral test will reduce stress and enhance enrichment by allowing animals opportunities for environmental exploration and by reducing the number of invasive experimental procedures. Two distinct behavioral approaches were tested: the obstacle-course test and temperament test. In the obstacle-course test, pigs were challenged (after an initial training period) to navigate a 10-object obstacle course; time and the number of collisions with the objects were recorded. In the temperament test, the time needed for pigs to complete 3 different tasks (human-approach, novel-object, and open-door tests) was recorded. The obstacle-course test revealed significant differences in time and number of collisions between swine with vision impairment and control animals, and the training period proved to be pivotal to avoid bias due to individual animal characteristics. In contrast, the temperament test was not altered by vision impairment but was validated to measure stress and behavioral alterations in laboratory pigs undergoing experimental procedures, thus achieving marked refinement of the study.

The roe deer is a seasonally breeding species with a reproductive cycle regulated by endogenous rhythms and photoperiod-sensitivity. Sexually mature bucks show hormonal and testicular activation during the reproductive season, with a peak in the rut period, and following gradual involution. Hair is a good matrix for non-invasive endocrinological analyses that provide long-term information without being influenced by the hormones&rsquo, pulsating release patterns in blood. The aim of the work was to quantify hair concentrations of testosterone and cortisol in wild roe deer bucks hunted during the pre- and post-rut period, using a radioimmunoassay methodology, and to look for differences between the two periods. The secondary objective was the evaluation of possible correlations of such hair concentrations with blood and morphometric parameters of the testes. Both hormones showed statistical differences, with opposing trends, when comparing the two periods: testosterone increased while cortisol decreased. The correlation analysis was in agreement with existing literature regarding metabolism/actions of these hormones and testicular morphometric parameters. This study represents the first report of the use of radioimmunoassay techniques to quantify testosterone and cortisol in roe deer hair, and may provide interesting insights into their reproductive physiology.

The roe deer (Capreolus capreolus) is a seasonal breeder, whose reproduction shows a distinct seasonal pattern with the breeding season (rut) lasting from mid-July to mid-August. Mature bucks show synchronized testicular cycles and transitions between totally arrested and activated spermatogenesis thanks to the peak of testosterone(TEST) between June and July [2]. Recently, hair has proven to be a good non-invasive matrix for endocrinological analyses, providing long-term information [1]. The aims of the present work were, firstly, to quantify hair concentrations of TEST and cortisol (CORT) in wild roe deer bucks hunted during pre and post rut period, using a radioimmunoassay (RIA) methodology, and to look for relevant differences between the two periods, and then to evaluate any possible correlation of such hair concentrations withblood ones and morphometric parameters of the testes. Twenty-eight mature roe deer bucks, were sampled during the hunting season between June 1st and July 15th 2017 (pre rut, n=14) and August 15th and September 30th 2017 (post rut, n=14) in the South-Western Bologna Apennines (Italy). All the animals analysed were killed and immediately transferred to the pertinent biometrical centre, where the personnel collected blood upon jugulation, hair from the dorsal caudal region of the animals and scrotums, including testis and epididymis. For the testicular analysis each testis was isolated, weighed and sectioned longitudinally. Major and minor axis were calculated and for each roe deer the measures were averaged between the two testis, in order to get a final mean volume. CORT and TEST were previously extracted from hair [1] and blood, and their concentrations were determined using a RIA technique. The results show that the testicular volume and weight significantly decreased in the post rut group (p=0.0089 and p=0.0056 respectively); the same happened for blood TEST concentrations (p=0.0008). The blood CORT levels did not show any statistical difference between the two groups (p=0.8336). The hair results showed an increase in TEST levels in the post-rut (p=0.00003), almost double the pre-rut ones. TEST reaches its blood peak in the pre-rut period, therefore it is likely that high hair post rut values are the direct reflection of high blood pre rut values. Differently, CORT levels decrease in the post-rut (p=0.0014), and this may be due to different environmental factors, leading to higher concentrations of hair CORT in the pre-rut. The correlations found by the statistical analyses were in agreement with the existing literature about the fluctuating concentrations of the two hormones in the blood throughout the year and the cyclic growth and reduction of the testis [2]. Moreover, this study represents a first validation for the use of a RIA technique to quantify both TEST and CORT in roe deer hair, confirming the use of this matrix as a good long-term information provider, and giving interesting insights on the physiology of reproduction of this species.

Simple Summary The red deer, also known as the royal deer or European deer, is an artiodactyl mammal belonging to the Cervidae family, widely diffused in almost all of continental Europe. At the beginning of autumn in the Northern Hemisphere, the mating season begins. The males of red deer, called stags, are synchronized with the females, called hinds; indeed, at the beginning of the mating season, they show a marked increase in testosterone to match the hinds’ estrus cycle. Gestation lasts about 230 days, so that calves are born in mid to late spring, the most favorable period for their survival. Scientific data on the reproduction physiology of this peculiar species in wild conditions are lacking, including hormonal variations during pregnancy. The present study describes mean levels of two critical hormones, cortisol and progesterone, in both blood and hair of wild pregnant red deer hinds. Correlation analysis confirmed how animals hunted in later phases of pregnancy have higher hair progesterone. Abstract The red deer (Cervus elaphus L., 1758) is one of the largest deer species in the world. Females are seasonal polyestrous, with negative photoperiod: the increase of the night peak of melatonin determines the secretion of GnRH and, therefore, LH and FSH. To date there is little information regarding the hormonal control during pregnancy for this species; this could be due to the difficulty of sampling wild subjects, while farmed animals’ hormonal concentrations may not reflect the physiology of the animal in a natural state. In this study we evaluated the concentration of cortisol and progesterone, extracted from blood and hair, on 10 wild and pregnant red deer females. Belonging to the population of the Bolognese Apennines (Italy), the hinds were sampled in the January–March 2018 period, according to the regional selective hunting plan. Plasma progesterone (P4) ranged from a minimum of 1.9 to a maximum of 7.48 ng/mL; while hair P4 concentrations varied from 41.68 to 153.57 pg/mg. The plasma and hair cortisol ranges are respectively 0.4–2.97 ng/mL and 0.03–0.55 pg/mg; the only significant correlation was found between hair concentration of P4 and the date of death. The results of this preliminary study represent a small step towards a better knowledge of this species’ physiology during pregnancy.

Porcine models of ophthalmological diseases are often used in pre-clinical translational studies due to pigs’ similarities to humans. In particular, the iodoacetic acid (IAA) model of photoreceptor degeneration seems to mimic well the endstage phenotype of human pathologies as retinitis pigmentosa and age-related macular degeneration, with high potential for prosthesis/retinal devices testing. IAA is capable of inducing photoreceptor death by blockage of glycolysis, and its effects on the retina have been described. Nonetheless, up to date, literature lacks of a comprehensive morpho-functional characterization of the entire visual system of this model. This gap is particularly critical for prosthesis testing as inner retinal structures and optic pathways must be preserved to elicit cortical responses and restore vision. In this study, we investigated the functional and anatomical features of the visual system of IAA-treated pigs and compared them to control animals. IAA was administered intravenously at 12 mg/kg; control animals received saline solution (NaCl 0.9% w/v). Electrophysiological analyses included full-field (ffERGs) and pattern (PERGs) electroretinograms and flash visually evoked potentials (fVEPs). Histological evaluations were performed on the retina and the optic pathways and included thickness of the different retinal layers, ganglion cells count, and immunohistochemistry for microglial cells, macroglial cells, and oligodendrocytes. The histological results indicate that IAA treatment does not affect the morphology of the inner retina and optic pathways. Electrophysiology confirms the selective rod and partial cone degeneration, but is ambiguous as to the functionality of the optic pathways, seemingly preserved as indicated by the still detectable fVEPs. Overall, the work ameliorates the characterization of such rapid and cost-effective model, providing more strength and reliability for future pre-clinical translational trials.

The Raphe Pallidus (RPa) is a region of the brainstem that was shown to modulate the sympathetic outflow to many tissues and organs involved in thermoregulation and energy expenditure. In rodents, the pharmacological activation of RPa neurons was shown to increase the activity of the brown adipose tissue, heart rate, and expired CO2, whereas their inhibition was shown to induce cutaneous vasodilation and a state of hypothermia that, when prolonged, leads to a state resembling torpor referred to as synthetic torpor. If translatable to humans, this synthetic torpor-inducing procedure would be advantageous in many clinical settings. A first step to explore such translatability, has been to verify whether the neurons within the RPa play the same role described for rodents in a larger mammal such as the pig. In the present study, we show that the physiological responses inducible by the pharmacological stimulation of RPa neurons are very similar to those observed in rodents. Injection of the GABAA agonist GABAzine in the RPa induced an increase in heart rate (from 99 to 174bpm), systolic (from 87 to 170mmHg) and diastolic (from 51 to 98mmHg) arterial pressure, and end-tidal CO2 (from 49 to 62 mmHg). All these changes were reversed by the injection in the same area of the GABAA agonist muscimol. These results support the possibility for RPa neurons to be a key target in the research for a safe and effective procedure for the induction of synthetic torpor in humans.

Antimicrobial resistance is increasing within the porcine industry with consequential high impact on human health, leading to a need for new antimicrobials. Lately, the scientific community has turned its interest towards natural compounds, and different essential oils have been tested on spermatozoa for preliminary assessment of toxicity before considering them as good substitutes for standard antibiotics. The aim of the present work was to investigate the potential antimicrobial effect of Melaleuca alternifolia and Rosmarinus officinalis essential oils, already evaluated for toxicity, on swine artificial insemination doses deprived of spermatozoa and stored at 16 °C for 5 days. This was accomplished by setting up an in vitro model with a standardized quantity of E. coli. Essential oils, previously chemo-characterized by means of gas chromatography, were tested at 0.2 and 0.4 mg/ml. Analyses, performed at 24 and 120 h, included optical density evaluation, bacterial DNA quantification by qPCR, and colony count. The results demonstrate that both Melaleuca alternifolia and Rosmarinus officinalis essential oils, at a concentration of 0.4 mg/ml, are capable of delivering similar effects to ampicillin, used as control, on the experimental samples. At the lower concentration, M. alternifolia essential oil seemed more effective when compared to R. officinalis. Overall, these findings strengthen the hypothesis of the potential use of phyto-complexes as antimicrobial agents for reproductive biotechnologies.

1-D codes are still much utilized regarding the preliminary design and the design phase of a single component. In particular, combustors is one of the most critical component in gas turbine engine and its design mainly affects all other components, starting from the turbine. During the initial phases of the design process, parameters are known approximately; during this phase, critical for the definition of a starting set of design parameters, there is little point in doing high fidelity Computational Fluid Dynamics (CFD) analyses. On the contrary, the exploration of the whole space is extremely important to better understand the behaviour of the system and to focus on the design objectives. Uncertainty quantification (UQ), mainly developed in recent years and applied in many fields, can be really helpful in the preliminary design phase and also as a support during the whole design process. This work comes with the idea to estimate the main source of geometrical uncertainties in the design phase of a combustor. The test case is based on a full annular lean burn combustor, tested at Central Institute of Aviation Motors (CIAM) during the LEMCOTEC (Low EMissions COre-engine TEChnologies) European project. Among the test points investigated in the experimental campaign, the Approach condition is analysed. The inner liner is considered to analyse the metal temperature. Therm-1D, a 1-D in-house simulation code, is used to model the combustor. After having built the baseline case of the combustor, several uncertainty analyses are investigated. In particular a classical Monte Carlo approach is compared with 4 innovative polynomial-chaos approaches for each group: Gauss quadrature, total order with Latin Hypercube sampling (LHS), probabilistic collocation and stochastic collocation. The analyses proved how the last two methods give the same results with a sensible lower amount of simulation (depending on the number of input variables). An additional sensitivity analysis to the order of the polynomial is conducted and results show how, with a 3rd order polynomial approximation, results can be very similar to those ones of the Monte Carlo simulation. Lastly, results are compared with experimental data to achieve a better understanding of the most relevant input parameters and the propagation of their uncertainty on the results.