National Projects

Tipology: Current Research 2021 (IZSLER 01/21 RC) Funding body: Ministero della Salute Period: 2021-2024 Head: Prof. Andrea Armani The project involves the development of an analytical procedure based on the metabarcoding technique that can be used by official laboratories for the identification of species in complex food matrices. Starting with a systematic review of the scientific literature on the applications of the metabarcoding technique in foods of animal origin, the molecular markers and primers of choice for metabarcoding analysis will be evaluated and subsequently tested.

Tipology: Current Research 2022 (IZS LT 14/22 RC) Funding Body: Ministero della Salute Periodo: 2022-2024 Head: Prof. Andrea Armani The aim of the project is to inform and protect the consumer and, in addition, to foster the proper development of e-commerce in food companies. Specifically, commercially relevant foodstuffs, potentially posing a health risk, sold online will be identified and subsequently analysed for the identification of the animal and plant species present, by means of DNA barcoding analysis based on next-generation sequencing technologies (Next Generation Sequencing-NGS).

Type: Current Research 2023 (IZS PB 02/23 RC) Funding body: Ministry of Health Period: 2023-2025 Head: Prof. Andrea Armani The aim of the work is the standardisation of molecular investigations based on metabarcoding, using Next Generation Sequencing (NGS), for the identification of species in meat products.

Type: University Research Project (PRA) 2022 (PRA_22_13) Funding body: University of Pisa Period: 2022-2024 Head: Prof. Andrea Armani The metabarcoding (MB) method based on NGS technologies will be applied to the identification of edible insect species in order to develop and validate an analytical procedure that can be transferred to official and private laboratories operating in the field of food safety for the analysis of food and feed. The MB will also be used to characterise the microbiome and microbiota of different edible insect species, also analysing the influence of breeding techniques. This process will make it possible to assess the presence of microorganisms that are pathogenic to humans and/or animals, confirming the results by means of traditional culture tests.

Typology: PRIN 2022 PNRR – Projects of Significant National Interest (PRIN), Ministry of University and Research MUR Research project: ERC subfields SH1_7 Behavioural economics; experimental economics; neuro-economics SH7_5 Sustainability sciences, environment and resources LS9_10 Veterinary and applied animal sciences Funding body: Ministry of University and Research MUR, Funds National Recovery and Resilience Plan (PNRR) Period: November 2023-November 2025 Responsible: UNIPI Unit Manager (Partner) – Dr. Simone Mancini The InsectFish project aims to promote sustainability in the fish sector by using insect meal in fish feed. The final objective is to investigate the quality, nutritional profile, sensory evaluation and consumer perception of fish fed with insect meal compared to a traditional fish (https://insectfish.unipr.it/).

Tipologia: PRIN: PROGETTI DI RICERCA DI RILEVANTE INTERESSE NAZIONALE – Bando 2022 Programma di ricerca: LS – Life Sciences Ente finanziatore: Ministero dell’Università e della Ricerca MUR. Periodo: Novembre 2023-Novembre 2025 Responsabile: Responsabile di Unità UNIPI (Partner) – Dott. Carlo Cantile L’obiettivo generale del presente progetto è lo studio delle più importanti malattie congenite e genetiche del sistema nervoso centrale (SNC) che negli ultimi decenni sono diventate motivo di grande preoccupazione per i veterinari e gli allevatori piemontesi. La diffusione di nuove malattie infettive ha provocato un aumento evidente, ma talvolta trascurato, delle malformazioni congenite, soprattutto del SNC. Inoltre, la maggiore pressione selettiva e la conseguente consanguineità dei bovini hanno portato a un aumento del rischio di difetti genetici. Le razze locali italiane hanno dato un enorme contributo alle malattie genetiche, poiché la limitata dimensione della popolazione di queste razze e l’uso su larga scala di un numero piuttosto limitato di tori di alto livello, finalizzato alla ricerca delle migliori caratteristiche produttive, ha aumentato la possibilità di stabilire alleli deleteri in questa popolazione.

Typology: National Projects – MASAF National Rural Development Programme 2014-2022 Research programme: PSRN 2014-2022 Sub-measure 10.2 – Support for the conservation, use and sustainable development of genetic resources in agriculture – Biodiversity/poultry Funding body: EAFRD 2014-2020 Period: 2021-2024 (this is the duration of the TuBAvI-2 project) Responsible: Margherita Marzoni Fecia di Cossato

The project stems from the association of Italian universities that have been committed for years to the conservation of biodiversity through the direct maintenance of native breed populations of the main poultry species of livestock interest. It is dedicated to the safeguard, conservation and valorisation of genetic resources in poultry farming, historically present in our country. It consists of several actions to provide knowledge and new tools to promote in situ and ex situ in vitro techniques for biodiversity conservation. The actions concern: phenotypic characterisation (Action 1), genetic characterisation (Action 2), estimation of conservation indices, mating plans and reproductive management (Action 4), evaluation of inbreeding and genetic diversity in populations and calculation of inbreeding (Action 5), monitoring and evaluation of genetic diversity in native breeds (Action 6), evaluation and identification of genetic resistance traits (Action 7) and information and dissemination actions (Action 10). UniPI’s specific project deals with the Gallus gallus species with studies focused on Action 1 (aimed at the core populations of the three breeds conserved in situ: Livorno, Siciliana and Ancona), Action 7 (aimed at eight breeds) and Action 10, as well as contributing to the implementation of research in the actions falling within the competence of the other Partners, providing the genetic material of its own conserved genetic cores with direct maintenance.

Typology: NRP Research programme: Spoke-10 ‘Population Health Funding body: European Union Period: 2022-2025 Responsible: Prof.ssa Elena Bargelli (coordinator), Prof. Francesco Paolo Di Iacovo, Prof.ssa Roberta Moruzzo Brief description of the project This project is coordinated by the SSSA and involves the partners UNIFI, UNIPI, UNISI, UNISTRASI whose proposals will contribute to provide methods and tools to support the governance of the health ecosystem and provide evidence to bridge the gap between innovation and large-scale implementation.

Typology: University Research Project (PRA) 2022 (PRA_22_13) Research programme: Antioxidants and pro-oxidants in veterinary medicine Funding body: University of Pisa Period: 2022-2024 Head: Mario Giorgi Short description of the Project: Cryopreservation of gametes is a technique widely used in animals and humans with the aim of preserving genetic material over time. In animals, this technique plays a key role in preserving biodiversity by creating banks of frozen genetic material, especially semen. During cryopreservation procedures, spermatozoa are subjected to various types of stress that often compromise their viability and fertility.In the horse, as in other animal species, countless substances have been added to spermatozoa cryopreservation media in order to reduce the damage associated with this technique (Oliveira et al., 2010; Del Prete et al., 2019; Silva et al., 2019). Horse spermatozoa membranes are composed of high levels of polyunsaturated fatty acids (PUFAs), which make them particularly sensitive to oxidative stress. Several studies have tried to add antioxidants to the media used for equine semen cryopreservation in order to mitigate the negative action of free radicals (ROS), but the results obtained have been variable and often disappointing (Doidar et al., 2018; Kumar et al., 2018; Lecewicz et al., 2019; Partyka et al., 2021; Tian et a., 2021). Recently, it has been shown that the exposure of semen to pro-oxidative substances (nitric oxide, ethanol, high hydrostatic pressure) is able to activate the intrinsic antioxidant system of the spermatozoon, improving its quality and resistance to degradation. Among these substances, ozone (O3), an unstable gas that is already used for various therapeutic purposes, has shown particular promise: its ability to induce controlled oxidative stress leads to an increase in the defence capacity of spermatozoa against the aggression of ROS formed during freezing (Pereira et al., 2022). Another approach to improving semen quality and its cryopreservability is based on the use of food supplements: numerous formulations of antioxidant principles and vitamins have been evaluated, in different animal species and in humans, with discordant results in terms of improving semen characteristics (Stradaioli et al., 2004). In particular, in horses, supplementation with PUFAs, carnitine, antioxidants, vitamins and botanical extracts has been investigated and led to controversial results (Da Silva Leitão Carvalho et al., 2020; Da Silva Leitão Peres et al., 2020).

Typology: PRIN 2022 PNRR – Projects of Significant National Interest (PRIN), Ministry of University and Research MUR. Research programme (operation 10.2.01 Conservation of animal and plant biodiversity) Funding body: Ministry of University and Research MUR, Funds National Recovery and Resilience Plan (PNRR) Period: November 2023-November 2025 Responsible: UNIPI Unit Manager (Partner) -Prof.ssa Francesca Bonelli

The livestock sector today faces several challenges, including the demand for less impact on the environment, improved animal welfare and less intensive production. A key intervention could be the improvement of animal health, reducing diseases and related losses, with the aim of making production more efficient and sustainable, in line with the ‘From Farm to Fork’ strategy promoted by the European Union. Recently, literature is exploring the potential use of colostrum as a nutraceutical to prevent disease in livestock species and limit the use of antimicrobials. However, there is still a long way to go. The overall aim of the project will be to evaluate, through a multi-omics approach of colostrum and blood, the potential nutraceutical use of colostrum in calves, foals and piglets. The project is divided into 4 work packages (WPs). WP1 and WP4 respectively represent the coordination of the two Research Units involved (UNIPI and UNIBO) and the creation of an inter-species database of the results obtained. WP2 and WP3, on the other hand, have the aim of evaluating, by means of a multi-omics analysis, the colostrum and blood characteristics in the three investigated species by correlating them during the colostral phase (WP2); the same subjects included in WP3 will then be divided into 3 treatment groups for the determination of the potential use of colostrum in the post-colostral phase as a nutraceutical to improve health, welfare and production.

Type: University research project Research programme: Physiology of ageing Funding body: University of Pisa Period: 2022-24 Head: Prof. Angelo Gazzano Brief description of the project The research aims to identify the presence of possible physiological, behavioural and genetic indicators of successful ageing in canine and equine species. As far as the dog is concerned, behavioural data will be collected in Labrador Retriever breed dogs, by means of the CADES card, to detect the presence of cognitive dysfunction. The pedigree of the subjects will be evaluated to calculate consanguinity and search for possible correlations with the degree of severity of cognitive impairment detected by the CADES card. A blood sample will also be taken from each animal for the determination of beta amyloid, tau protein and light neurofilament in order to highlight a correlation between these indices of neuronal degeneration and the severity of cognitive degeneration identified by the card.An aliquot of blood will be used for genotyping with SNP (Single Nucleotide Polymorphism) genomic markers in order to identify senescence-related genes and a further aliquot will be used to search for blood parasites. With regard to the equine species, behavioural and management data will be collected through the use of special questionnaires, distributed to breeders and owners of Arabian horses, to detect the possible occurrence and frequency of particular physical and behavioural problems and organic pathologies. The animals will be subdivided according to age and, through the identification document, the calculation of consanguinity will be carried out to detect correlations with possible behavioural and physical problems and diseases related to senescence. A sample of 10 horses over the age of 20 years will have a blood sample taken that will be used for the validation of commercial kits for the determination of beta amyloid and tau protein.

Typology: PRIN 2022 PNRR – Projects of Significant National Interest (PRIN), Ministry of University and Research MUR Research programme: LS – Life sciences, SH – Social sciences and humanities Funding body: Ministry of University and Research MUR, Funds National Recovery and Resilience Plan (PNRR) Period: November 2023-November 2025 Responsible: UNIPI Unit Manager (Partner)- Dr. Francesco Riccioli Short description of the Project Aquaculture and edible insect production have recently been proposed as solutions to ensure access to safe and sustainable food without burdening natural resources and biodiversity, while mitigating the impact of climate change. Despite their high nutritional value and sustainability, Italians are reluctant to consume farmed fish and insects.

Typology: Regional call Research programme: Improvement of the beekeeping sector through innovative technologies Funding body: Submeasure 16.2, RDP 2014-2022 of the Region of Tuscany Period: 2022-2024 months Responsible: Head of O.U. Mina Martini Short project description: The project is aimed at improving beekeeping production (reduction of production costs, quality improvement of productions, market enhancement) through technological improvement.

Typology: University project Research programme: University Research Projects (PRA) 2022-2023 Funding body: University of Pisa Period: 2022-2024 Responsible: Prof. Valentina Meucci Short project description: The prudent and appropriate use of antibiotics includes measures to maximise the efficacy of these drugs and limit the development and spread of resistance. Actions to promote the prudent use of antibiotics have multidisciplinary characteristics and are intended to provide tools for the appropriate prescription of antimicrobial drugs and, more generally, for the overall management (prevention, diagnosis and treatment) of subjects presenting an infection of bacterial origin. This research project, addressed to the equine species, aims: 1) to retrospectively evaluate the antibiotic prescriptions of veterinary hospitals and clinics participating in the project to estimate their adherence to the criteria of prudent use; 2) to evaluate the possible use of blood biomarkers (BM) (procalcitonin and oxidative stress markers) in equine clinical practice to guide the rational use of antibiotics. The results obtained from the retrospective analysis of antibiotic prescriptions will be disseminated to health professionals in the area; BMs will be proposed to guide and monitor antibiotic therapy

Typology: Current research projects 2022 Research programme: Animal welfare Funding body: Ministry of Health Period: 2022-2024 Responsible: Head of O.U. Mina Martini Short project description: The project aims to assess the welfare level of sheep farms in Lazio and Tuscany, in relation to grazing possibility and nutritional intake, going to evaluate the respective quanti-qualitative productions and farm economic yield. This information will contribute to improve the knowledge on the relationship between the welfare level and the grazing usability, correlating them with the production and economic parameters of the sheep milk sector farms.

Typology: University Research Projects Research programme: not applicable Funding body: University of Pisa Period:2022 -2024 Responsible: Prof. Federica Salari Short description of the project The sheep sector accounts for just over 1% of the value of national agricultural production, determining in 2019 a total of approximately 624 million euros; of which 460 million generated by the milk sector (ISMEA, 2020). The national sheep herd is mainly concentrated in a few regions: Sardinia (47%), Sicily (11%), Lazio (9%) and Tuscany (5%) (BDN, 2021). Tuscany is therefore among the most important regions for the sheep sector and the sector plays an important role with a 3% incidence on the total value of regional agriculture (ISMEA, 2020).In Tuscany there are 3,770 sheep farms and more than 80 per cent of the animals are geared to milk production (BDN, 2021). Researchers estimate that globally small ruminants are responsible for about 6.5% of livestock sector emissions (475 million tonnes of CO2 eq) (FAO, 2016). Sheep contribute less to these emissions than other livestock species (FAO, 2022), however, since climate change must remain within acceptable limits, each individual production sector must reduce its carbon footprint and, more generally, its environmental impact (O’Mara, 2011). In recent years, Life Cycle Assessment (LCA) has emerged as an internationally accepted method for assessing the environmental impact of a product. The application of LCA to livestock production systems is a relatively new area of research (Cottle and Cowie, 2016), there is therefore a need for studies that analyse and enable estimates of the environmental impacts of sheep farming (Akul and Bassim, 2021). For what has been said, the aim of the study will be to quantify the potential environmental impact of sheep milk production in Tuscany and to identify the critical points on which it is necessary to act in order to increase its sustainability. The assessment of the environmental impact of the farms will be carried out through the Life Cycle Assessment methodology, the data necessary for the analysis will be collected on site through questionnaires to the farmers and will then be processed through the use of a specific software; the results will be disseminated through seminars, focuses, communications and publications on international level and popular magazines of the sector.

Typology: Ministry of Health project Research programme: Ministry of Health call with a specific objective Funding body: Ministry of Health Period: 2022-2024 Responsible: National Dr. Cristina Roncoroni (IZS) UNIPI Prof. Luigi Intorre Short description of the project The Departments of Veterinary Sciences and Pharmacology will be engaged in the preparation of a pharmaceutical form of GonaCon for oral use based on mucoadhesive functional excipients, stabilisers of labile compounds (proteins and peptides) and promoters of intestinal absorption and its loading in a gastro-resistant form.

Typology: PRIN 2022- Projects of Significant National Interest (PRIN), Ministry of University and Research MUR Research programme: LS – Life Sciences Funding body: Ministry of University and Research MUR Period: November 2023-November 2025 Responsible: Head of Unit UNIPI (Partner)- Prof.ssa Francesca Millanta Short description of the project The aim of the project is to collect and establish primary 3D cell cultures from canine spontaneous neoplasms, by a small and handheld bioreactor developed to recreate a 3D tissue-like structure in a closed system, which integrates a 3D scaffold able to sustain growth of normal and pathological cells within a biologically relevant environment. Two OUs (Milan and Pisa) will provide samples of osteosarcomas and splenic hemangiosarcomas (Milan), mucosal melanoma and urothelial carcinoma (Pisa) from canine patients and will evaluate their immunohistochemical phenotype. The OU in Bologna will generate the 3D cultures from the same cases, part of which will be formalin fixed, paraffin embedded and returned to Milan and Pisa OU to comparatively perform the same immunohistochemical exam as for the primary tumor. At the Bologna OU genotypic and biochemical investigations will also be performed both on samples from primary tumours and 3D cultures. The final goals are the demonstration of: 1) expression of the same differentiation markers (CD31 and FVIII-RA for hemangiosarcomas, Vimentin, RUNX2 and ALP for osteosarcomas; Melan-A, S100 and PNL2 for melanomas, PANcytokeratin and uroplakin for urothelial carcinomas) by neoplastic cells in the primary tumor and the corresponding 3D system; 2) expression of the same genetic alterations by Next Generation Sequencing (NGS) of a selection of genes (BRAF, CDKN2A, DMD, KRAS, MYC, NRAS , PIK3CA, PLCG1, PTEN, RB1, SETD2, TP53) in the primary tumor and the corresponding 3D system, which, integrated with results from point 1, will increase the evidence of the replication, in the 3D system, of the same primary neoplastic cells; MUR – BANDO 2022 Ministero dell’Università e della Ricerca 3) identification of metabolic switch of cancer cells in primary tumours and 3D cultures from oxidative to glycolytic metabolism to counteract the mitochondrial OXPHOS deficiency and evaluation of the molecular events of mitochondrial dysfunction that trigger the permeability transition pore opening during programmed cell death in cancer cells. The dog is a good candidate species because of a well-established literature with consistent data, providing a model with higher probability to bear genetic anomalies in the primary tumor that may be demonstrated specifically in the corresponding 3D culture. Furtheremore two out of the four neoplastic types chosen (osteosarcoma and mucosal melanoma) are spontaneous animal models of human diseases and, by this way, the proposal provides data of translational medicine. The main point of advancement of knowledge is the identification and implementation of available protocols for setting up 3D systems capable of producing in a short time (7 days) primary 3D cell cultures of tumors that maintain the morphological, phenotypic, genetic and metabolic characteristics of the primary neoplasm. A field of application of the results provided is to employ the proposed model for more accurate and rapid studies of targeted therapies.