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Animal welfare is a growing, compelling and urgent topic because of the interest that it generates among the citizens and consumers. The goal to be fulfilled is to define systems and protocols for assessing animal welfare that should be impartial, reproducible and science-based on risk analysis. On these premises, the Italian Ministry of Health has signed an agreement with the IZSLER, through the CReNBA and IZSM through the CReNBuf on development and management of a system to evaluation of welfare and biosecurity in buffaloes breeding system. The checklist development was based on the Risk Assessment Methodology as suggested by EFSA. Building on the CReNBA’s work developed on the welfare dairy cow, this method is based on the analysis of two data groups: The assessment of the hazards (non-ABMs) occurring as a result of environmental conditions; the assessment of the risks, with the concerned adverse effects (ABMs), run by animals living in those environments. The system developed consists in 83 observations, listed in a multiple-choice checklist divided in five macro-areas: Farm management and personnel, Facilities and equipment, Animal based measures, Biosecurity and Alarm systems. The result of each area also provides an indication of the burden and importance of each of these on the final calculation of the animal welfare value. These checks represent a functional and smart instrument to allow assign a numerical animal welfare index to each farm. In addition, by the data collected in each area, to supply at the Veterinarians and breeders the tools to improve farm management and structures, respecting the farm’s sustainability, and is preparatory to given to the development of a Ministerial trademark for animal welfare, giving answers to consumers and add value to the correct activities of the farmers.

Hyaluronic acid (HA) is a key component of synovial fluid as it plays a crucial role in joint physiology. Its biological activity is influenced by molecular weight, local concentration, and persistence in joints. High-molecular-weight HA has a consolidated history of clinical use, whereas little is known about the metabolic effect of low-molecular-weight hyaluronate on cartilage differentiation. This study explores the potential of HA-based nanoparticles (NPs) on chondrocytes differentiation in vitro. Starting from 25 kDa and 250 kDa sodium hyaluronate solutions, two types of NPs were prepared by antisolvent precipitation in ethanol. The resulting NPs were dried in the presence of dipalmitoyl phosphatidylcholine, a natural synovial fluid component, then applied on an in vitro model of horse articular chondrocytes: no toxicity was observed and NPs prepared from 250 kDa HA promoted chondrocyte differentiation to a larger extent with respect to corresponding HA solutions, as evidenced by increased gene expression of chondrogenic markers ( Col2a1 and Sox9 ) and reduced expression of dedifferentiation markers ( Col1a1 and Runx2 ). These findings suggest that HA-based NPs are more effective at promoting the cellular internalization of the molecule and the differentiation of chondrocytes in vitro and could be a promising platform for drug delivery and cartilage repair.

Idiopathic pulmonary fibrosis (IPF) is a chronic inflammatory and fibrotic pathological condition with undefined effective therapies and a poor prognosis, partly due to the lack of specific and effective therapies. Galectin 3 (Gal-3), a pro-fibrotic ß-galactoside binding lectin, was upregulated in the early stages of the pathology, suggesting that it may be considered a marker of active fibrosis. In the present in vitro study, we use Hylach ® , a lactose-modified hyaluronic acid able to bind Gal-3, to prevent the activation of lung myofibroblast and the consequent excessive ECM protein cell expression. Primary human pulmonary fibroblasts obtained from normal and IPF subjects activated with TGF-β were used, and changes in cell viability, fibrotic components, and pro-inflammatory mediator expression at both gene and protein levels were analyzed. Hylach compounds with a lactosylation degree of about 10% and 30% (Hylach1 and Hylach 2), administrated to TGF-β-stimulated lung fibroblast cultures, significantly downregulated α-smooth muscle actin (α-SMA) gene expression and decreased collagen type I, collagen type III, elastin, fibronectin gene and protein expression to near baseline values. This anti-fibrotic activity is accompanied by a strong anti-inflammatory effect and by a downregulation of the gene expression of Smad2 for both Hylachs in comparison to the native HA. In conclusion, the Gal-3 binding molecules Hylachs attenuated inflammation and TGF-β-induced over-expression of α-SMA and ECM protein expression by primary human lung fibroblasts, providing a new direction for the treatment of pulmonary fibrotic diseases.

Background : The best application modality of high-fidelity simulation in graduate critical care nursing courses is still rarely investigated in nursing research. This is an important issue since advanced nursing skills are necessary to effectively respond to critically ill patients' care needs. The aim of the study was to examine the influence of a modified teaching model based on multiple exposures to high-fidelity simulations on both the learning outcomes and the perceptions of graduate students enrolled in a critical care nursing course. Methods : A multimethod study involving a sample of graduate critical care nursing students was conducted. A theoretical teaching model focused on multiple exposures to high-fidelity simulations is currently applied as a teaching method in an Italian critical care nursing course. According to the Kirkpatrick model for evaluating training programs, the performance, self-efficacy, and self-confidence in managing critically ill patients were considered learning outcomes, while satisfaction with learning and students' lived experiences during the experimental phases were considered students' perceptions. Results : Multiple exposures to high-fidelity simulations significantly improved performance, self-efficacy, and self-confidence in managing virtual critically ill patients' care needs. The satisfaction level was high, while lived experiences of participants were positive and allowed for better explanation of quantitative results of this study. Conclusions : Multiple exposures to high-fidelity simulations can be considered a valuable teaching method that can improve the learning outcomes of graduate nurses enrolled in an intensive care course.

Judicious antimicrobial stewardship in livestock industry is needed to reduce the use of antimicrobials (AMU) and the associated risk of antimicrobial resistance. Biosecurity measures are acknowledged for their role against the spread of diseases and the importance in reducing AMU in different species. However, their effectiveness in beef production has been scarcely considered. The aim of this study was to investigate the effect of the quarantine strategy on AMU in beef cattle. A total of 1206 Charolaise animals in five farms were included in the trial. Roughly half of the animals followed the standard procedure of the fattening cycle (no-quarantine; NO-QUA group) and half followed a 30-day period of quarantine (QUA group) since their arrival. Performance and antimicrobial data were recorded and a treatment incidence 100 (TI100it) per animal was calculated. Penicillins was the most used class of antimicrobials. Differences between groups were significant for males only, with NO-QUA group having greater TI100it (3.76 vs. 3.24; p < 0.05) and lower body weight at slaughter (713.4 vs. 723.7 kg; p < 0.05) than QUA group. Results suggest that quarantine strategy can reduce AMU in males without compromising their performance, whereas further investigation is needed for females.

Aim: To evaluate the University of Pavia students about physical activity (PA) changes before, during and after the COVID-19 pandemic., Methods: The International Physical Activity Questionnaires (IPAQ) survey was employed to evaluate the PA in three periods: the pre-pandemic period, during national stay-at-home order (March 9th - May 4th 2020), current PA. Exercise intensity for each period was defined using the Metabolic Equivalent of Task (Met) as unit of measurement. The questionnaire was administered online to university students from June 9th to July 4th 2021, structured in four sections, also collecting demographic data., Results: 55,6% of the study population reported a significant decrease in PA during lockdown. The number of active/very active subjects dropped from 72.2% in pre-pandemic period to 29.6% during containment measures. 50% reported a substantial increase in moving out of the lockdown. Stay-at-home order was associated with an increase in sedentary lifestyle (68.5%), which sharply decreased moving out from lockdown (two-third of study population). Average time in minutes spent sitting was 612 before pandemic, 844 during the pandemic and 670 after social restrictions., Conclusions: Lockdown had a negative impact on PA among the university students leading to an increase in sedentary behaviours. Following the gradual relaxation of the restrictive measures, situation has improved, without however returning to pre-pandemic level. It is of fundamental importance to study new strategies to promote healthy lifestyles while coping with the on-going pandemic.

The delivery of a dexamethasone formulation directly into the lung appears as an appropriate strategy to strengthen the systemic administration, reducing the dosage in the treatment of lung severe inflammations. For this purpose, a hyaluronic acid-dexamethasone formulation was developed, affording an inhalable reconstituted nanosuspension suitable to be aerosolized. The physico-chemical and biopharmaceutical properties of the formulation were tested: size, stability, loading of the spray-dried dry powder, reconstitution capability upon redispersion in aqueous media. Detailed structural insights on nanoparticles after reconstitution were obtained by light and X-ray scattering techniques. (1) The size of the nanoparticles, around 200 nm, is in the proper range for a possible engulfment by macrophages. (2) Their structure is of the core-shell type, hosting dexamethasone nanocrystals inside and carrying hyaluronic acid chains on the surface. This specific structure allows for nanosuspension stability and provides nanoparticles with muco-inert properties. (3) The nanosuspension can be efficiently aerosolized, allowing for a high drug fraction potentially reaching the deep lung. Thus, this formulation represents a promising tool for the lung administration via nebulization directly in the pipe of ventilators, to be used as such or as adjunct therapy for severe lung inflammation.

The concept of animal welfare (AW) has many meanings. Traditionally, AW has been considered as freedom from disease and suffering. Nowadays, growing attention goes to the concept of "positive animal welfare" (PAW), which can be interpreted within the concept of quality of life (QoL), thinking about a "balance of positives over negatives" and a "life worth living". In this vision, where the QoL represents a continuum between positives and negatives, the Italian National Reference Centre for Animal Welfare (CReNBA), within the Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), has developed a welfare assessment protocol for dairy cows, heifers, and calves in loose housing systems, including both animal-based and non-animal-based indicators, in which not only hazards but also benefits are identified. This protocol is part of an integrated monitoring system called "ClassyFarm", belonging to the Italian Ministry of Health and developed by IZSLER. The aim of this paper is to extrapolate from the mentioned protocol, a list of 38 best farming practices (on managerial and equipment factors) for ensuring a high level of welfare in dairy cattle. All stakeholders (veterinarians, farmers, competent authorities, consumers, etc.) can benefit of these best practices as a guide or toolbox to ensure a life worth living for these animals.

Information regarding the relationship between animal welfare (AW) and antimicrobial use (AMU) in dairy cows is limited. The current study aimed to investigate this relationship on Italian farms and to identify potential targets of AMU reduction. The study was performed at 79 Italian dairy farms housing over 15,000 cows during 2019. AW was scored with an on-farm protocol assessing farm management and staff training, housing systems, and animal-based measures. AMU was estimated using a defined daily dose per kg of animal biomass (DDDAit/biomass) for Italy. The median AW score was 73% (range: 56.6-86.8%). The median AMU was 4.8 DDDAit/biomass (range: 0-11.8). No relationship between the total AMU and AW was found. Management and staff training were positively associated with the use of the European Medicines Agency's category B antimicrobials, which are critical for human medicine, and with intramammary products for dry cow therapy. In those farms, antimicrobial stewardship should aim to reduce the category B antimicrobials and selective dry cow therapy. Our results underline the importance of implementing both an integrated monitoring system (AW, AMU, etc.) and antimicrobial stewardship tailored to the specific needs of each dairy farm.

Background: In postgraduate intensive care nursing courses, high-fidelity simulation is useful to prepare students to guarantee safe and quality care of critically ill patients. Surprisingly, this issue has not attracted sufficient attention in the literature, and it is not clear whether the linear application of the traditional high-fidelity simulation method based on prebriefing, the simulation session and debriefing, can serve as empirical reference in postgraduate students' education. The aim of this study was to investigate the lived experiences of postgraduate students receiving multiple exposures to an innovative high-fidelity simulation design based on Kolb's Experiential Learning Theory., Methods: A phenomenological study was conducted at an Italian University involving a purposive sample of 15 nursing students attending the postgraduate intensive care course. Audio-recorded face-to-face in-depth interviews were held by a researcher in a dedicated room complemented with non-verbal communication outlined in the field notes. Thematic analysis was used to analyse the transcribed data., Results: Three themes and ten categories were derived from the data analysis. The themes included pragmatic learning experience, the emotional path, and confidence., Conclusions: Multiple exposure to high-fidelity simulation was lived as a pragmatic learning experience enhancing the students' ability to apply theory into practice. This novel approach also contributed to the transition from negative to positive feelings and improved students' confidence about technical and non-technical skills when caring for a critically ill patient., (© 2021. The Author(s).)

The search for best performing carriers for dry powder inhalers is getting a great deal of interest to overcome the limitations posed by lactose. The aerosolization of adhesive mixtures between a carrier and a micronized drug is strongly influenced by the carrier solid-state properties. This work aimed at crystallizing kinetically stable D-mannitol polymorphs and at investigating their aerosolization performance when used in adhesive mixtures with two model drugs (salbutamol sulphate, SS, and budesonide, BUD) using a median and median/high resistance inhaler. A further goal was to assess in vitro the cytocompatibility of the produced polymer-doped mannitol polymorphs toward two lung epithelial cell lines. Kinetically stable (up to 12 months under accelerate conditions) α, and δ mannitol forms were crystallized in the presence of 2% w / w PVA and 1% w / w PVP respectively. These solid phases were compared with the β form and lactose as references. The solid-state properties of crystallized mannitol significantly affected aerosolization behavior, with the δ form affording the worst fine particle fraction with both the hydrophilic (9.3 and 6.5%) and the lipophilic (19.6 and 32%) model drugs, while α and β forms behaved in the same manner (11-13% for SS; 53-58% for BUD) and better than lactose (8 and 13% for SS; 26 and 39% for BUD). Recrystallized mannitol, but also PVA and PVP, proved to be safe excipients toward lung cell lines. We concluded that, also for mannitol, the physicochemical properties stemming from different crystal structures represent a tool for modulating carrier-drug interaction and, in turn, aerosolization performance.

The need for animal welfare definition and assessment is increasing worldwide, and several studies have been conducted to help fill the knowledge gaps regarding the welfare of cattle. However, further studies are needed to provide valid synthetized measures for welfare evaluation. The aim of this study was to assess the welfare status of 16 Sardinian dairy cattle farms, based on the developed Animal Welfare and Biosecurity Evaluation checklist (AWB-EF) and the corresponding hematological, biochemical, and electrophoretic profiles of these animals. Considering the AWB-EF as gold standard, blood samples were collected from 230 Holstein breed dairy cattle, aged between 3 and 8 years, out of the periparturient period, and with no clinical signs of specific pathologies. Principal Component (PC) and correlation analyses were performed to simplify phenomena interpretation and assess positive/negative associations. Four PCs were able to explain 76% of the total variability, and six laboratory parameters were strongly associated with the AWB-EF score (Spearman's correlation coefficient ≥ 0.40, p -Value < 0.05), reflecting the real health status of the animals. Given the complexity of animal welfare as a multidimensional concept and the need to include both animal-based and non-based measures in welfare evaluation, the present work represents a sound basis for future evaluation and veterinary health planning.

Antimicrobial use (AMU) in livestock species and the associated antimicrobial resistance are a global concern, thus strategies for their reduction and a more judicious use are needed. Previous research has revealed a link between improved animal welfare, biosecurity and AMU reduction in pig and dairy sectors, however, little is known about the beef sector. This study aimed to investigate the impact of welfare standards and biosecurity on AMU in beef cattle. Data on performance traits and AMU were collected over a 3.5 year time from 27 specialised beef farms and a treatment incidence was calculated using the defined daily dose for animals. An on-farm assessment was carried out by assigning a score from 0 (very poor) to 100% (very good) to 3 sections: welfare, biosecurity and emergency management. The highest average score was obtained for the welfare section (76%) followed by emergency management (39%) and biosecurity (24%). This suggests that major focus on strategies for the implementation of biosecurity measures and emergency management is needed, due to the low scores reported. A statistically significant lower AMU was observed with improved level of welfare. These results may be helpful for farm benchmarking and highlight the importance of improved animal welfare for an efficient antimicrobial stewardship.

Aspergillus flavus is a well-known ubiquitous fungus able to contaminate both in pre- and postharvest period different feed and food commodities. During their growth, these fungi can synthesise aflatoxins, secondary metabolites highly hazardous for animal and human health. The requirement of products with low impact on the environment and on human health, able to control aflatoxin production, has increased. In this work the effect of the basidiomycete Trametes versicolor on the aflatoxin production by A. flavus both in vitro and in maize, was investigated. The goal was to propose an environmental loyal tool for a significant control of aflatoxin production, in order to obtain feedstuffs and feed with a high standard of quality and safety to enhance the wellbeing of dairy cows. The presence of T. versicolor, grown on sugar beet pulp, inhibited the production of aflatoxin B1 in maize by A. flavus. Furthermore, treatment of contaminated maize with culture filtrates of T. versicolor containing ligninolytic enzymes, showed a significant reduction of the content of aflatoxin B1.