PhD in Veterinary Sciences for Animal Health and Food Safety

Contacts

Supervisor

Eugenio Martignani

Curriculum vitae

Curriculum Vitae

Phd thesis

Application of circulating-miRNAs expression profiles in an endangered Piedmontese sheep breed during the oestrous cycle

The conservation of local, rare, and well-adapted livestock breeds is essential for maintaining ecosystem services and ensuring resilience to harsh environments. MicroRNAs (miRNAs) are small, non-coding RNA molecules involved in post-transcriptional cellular regulation and could serve as biomarkers for physiological conditions. Produced in tissues, when they circulate in extracellular fluids, they are called circulating miRNAs (c-miRNAs). This PhD project focused on the endangered Italian breed Frabosana-Roaschina, investigating reproductive function and physiological stress responses in two trials with twenty ewes each. The first trial involved nulliparous ewes, while the second included pluriparous ones. Blood, saliva, and faeces were collected for two months after synchronization via progesterone sponges. Animals were also recorded throughout the sampling period for behavioural evaluation. Behavioural observations, hormonal analysis (oestradiol, progesterone, cortisol), and ultrasound scans were used to determine oestrus cycle phases. Two key moments were selected: the beginning of the follicular and luteal phases. Plasma c-miRNAs were sequenced, followed by enrichment analysis and differential expression analysis between the two phases. Since no acute stress moments were observed in Frabosana-Roaschina, a second study was conducted on the Spanish breed Rasa Aragonesa to explore miRNAs' role in short-term stress response. Fourteen lambs underwent transport, a common management practice, and their physiological responses (heart rate and body temperature) were monitored using subcutaneous bio-loggers. Moreover, salivary cortisol was estimated to detect the acute stress moment. The unloading phase was identified as the most stressful moment. In addition, c-miRNAs in saliva and plasma were analyzed before and after transport to assess stress-related changes. The PhD project identified c-miRNAs potentially involved in the regulation of the oestrus cycle and stress response in sheep. These findings contribute to understanding local breed adaptation and resilience, improving the reproductive performance and welfare of Frabosana-Roaschina, supporting conservation efforts. Additionally, the study highlighted the dynamic nature of ovine c-miRNAs in transport-induced stress in Rasa Aragonesa. In conclusion, this research investigates circulating miRNAs as potential biomarkers for reproductive welfare assessment and acute stress adaptation, contributing to livestock management strategies for endangered and well-adapted local breeds.

Research activities

Period Abroad & Courses

Facultad de Veterinaria, Universidad de Zaragoza (UNIZAR) with supervisor Dr. Genaro C. Miranda de la Lama (4 months)

Prizes and awards

Subject Expert in Veterinary Physiology – Verbale n.21 del 15 dicembre 2021 del Consiglio di Dipartimento di Scienze Veterinarie dell’Università degli Studi di Torino.

SISVET Young Researchers Award - June 2024

 

International Congress attendance:

• Manenti I., Miretti S., Viola I., Canto F., Toschi P., Bjarnason A., Miranda de la Lama G., Abecia J.A. Heart rate variability measured by subcutaneous bio-loggers during one day of transport in fattening lambs, IEEE 2024 INTERNATIONAL WORKSHOP ON MEASUREMENTS AND APPLICATIONS IN VETERINARY AND ANIMAL SCIENCES, 22^nd-24^th April 2024, Turin, Italy
• Manenti I., Abecia J.A., Viola I., Canto F., Toschi P., Bjarnason A., Accornero P., Martignani E., Miretti S. Transport and stress: bio-loggers and miRNAs to measure the adaptive physiological response in lambs, IEEE 2024 INTERNATIONAL WORKSHOP ON MEASUREMENTS AND APPLICATIONS IN VETERINARY AND ANIMAL SCIENCES, 22^nd-24^th April 2024, Turin, Italy
• Cappone E.E., Bongiorno V., Fiorilla E., Gariglio M., Macchi E., Manenti I., Stoppani N., Soglia D., Schiavone A., Measuring ethological parameters and their correlation with corticosterone levels in three slow-growing Piedmontese chicken breeds, IEEE 2024 INTERNATIONAL WORKSHOP ON MEASUREMENTS AND APPLICATIONS IN VETERINARY AND ANIMAL SCIENCES, 22^nd-24^th April 2024, Turin, Italy
• Manenti I., Abecia J. A., Viola I., Canto F., Toschi P., Bjarnason A., Miranda-de la Lama G., Miretti S. Effect of transport on body temperature and heart rate of lambs measured by subcutaneous bio-loggers, 8^th International Bio-Logging Science Symposium, 4^th-8^th March 2024, Tokyo, Japan
• Manenti I., Viola I., Ala U., Macchi E., Baratta M., Martignani E., Miretti S. Adaptation response in sheep: ewes in different cortisol clusters reveal changes in the expression of salivary miRNAs, 5° Annual Meeting of the European Veterinary Congress of Behavioural Medicine and Animal Welfare, 19^th-20^th October 2023, Pisa, Italy
• Viola I., Manenti I., Accornero P., Baratta M., Toschi P. mTOR signalling pathway as a key-modulator in placenta development: cell functionality and gene expression of trophoblast adaptive response during the early stage of pregnancy in sheep, 18th European Placenta Group IFPA meeting. 25^th-27^th November 2022, Paris, France

 

National Congress attendance:

• Manenti I., Ala U., Viola I., Miretti S., Macchi E., Baratta M., Martignani E. The circulating microRNAs enter the reproductive landscape of a threatened sheep breed: a sequencing data analysis, 77° Convegno SISVET, 12^nd-14^th June 2024, Parma, Italy
• Manenti I., Abecia J.A., Viola I., Canto F., Toschi P., Bjarnason A., Accornero P., Miretti S. Molecular biomarkers and subcutaneous bio-loggers to assess the lambs' adaptive physiological response during transport, 77° Convegno SISVET, 12^nd-14^th June 2024, Parma, Italy
• Viola I., Accornero P., Manenti I., Miretti S., Baratta M., Toschi P. mTOR is an essential gate in adapting the functional response of ovine trophoblast cells under stress-inducing environments, 77° Convegno SISVET, 12^nd-14^th June 2024, Parma, Italy
• Viola I., Sosa C., Canto F., Manenti I., Miretti S., Accornero P., Abecia J.A., Toschi P. Exogenous melatonin strengthens fetal-maternal cross-talk by ameliorating uterine microenvironment during the early stage of pregnancy in sheep, 77° Convegno SISVET, 12^nd-14^th June 2024, Parma, Italy
• Manenti I., Viola I., Ala U., Macchi E., Baratta M., Martignani E., Miretti S. The extreme phenotypes of cortisol levels reveal different expression patterns of miRNAs in sheep saliva, 76° Convegno SISVET, 21^st-23^rd June 2023, Bari, Italy
• Viola I., Manenti I., Accornero P., Baratta M., Toschi P. Autophagy effect on trophoblast cell functionality during early placenta development in sheep, 76° Convegno SISVET, 21^st-23^rd June 2023, Bari, Italy
• Manenti I., Macchi E., Baratta M., Miretti S., Viola I., Toschi P., Cornale P. Does hormonal profile influence behaviors in pubescent ewes?, 75° Convegno SISVET, 15^th-18^th June 2022, Lodi, Italy

 

Publications:

• [ACCEPTED] Manenti I., Sicuro B., Macchi E., Ponzio P., Crosetto R., Cavallari S., Tarantola M. and Mugnai C. Effect of different levels of environmental enrichment on the welfare of guppy (Poecilia reticulata). 2025, Frontiers in Fish Sciences, 3. doi: 10.3389/frish.2025.1581493
• Abecia J.A., Manenti I., Viola I., Toschi P., Palacios C., Canto F. and Miretti S. Triaxial accelerometers and subcutaneous biologgers as tools to determine diurnal and nocturnal changes in locomotor activity, body temperature, heart rate, and heart rate variability, after a melatonin treatment in lambs. 2025, Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology, 305, 111849. doi: 10.1016/j.cbpa.2025.111849
• Abecia J.A., Canto F., Viola I., Manenti I., Toschi P. and Miretti S. Weaning reduces body temperature and heart rate, and increases heart rate variability in ewes. 2025; Smart Agricultural Technology, p.100869. doi: 10.1016/j.atech.2025.100869
• Bongiorno V., Fiorilla E., Gariglio M., Zambotto V., Cappone E.E., Bergagna S., Manenti I., Macchi E., Gai F. and Schiavone A. New horizons in live and dehydrated black soldier fly larvae usage: Behavioral and welfare implications in “Bianca di Saluzzo” cockerels. 2025; PloS one, 20(2), p.e0318793. doi: 10.1371/journal.pone.0318793
• Manenti I., Ala U., Macchi E., Viola I., Toschi P., Accornero P., Baratta M., Miretti S., Martignani E. Circulating-miRNAs expression profiles in an endangered Piedmontese sheep breed during the estrus cycle. 2024; Frontiers in Veterinary Sciences, 11, p.1458463, doi: 10.3389/fvets.2024.1458463
• Viola I., Accornero P., Manenti I., Miretti S., Baratta M., Toschi P. mTOR is an essential gate in adapting the functional response of ovine trophoblast cells under stress-inducing environments. 2024; Placenta, 19; 158:14-22. doi: 10.1016/j.placenta.2024.09.011
• Viola I., Sosa C., Accornero P., Manenti I., Canto F., Miretti S., Abecia J.A., Toschi, P. Exogenous melatonin ameliorates embryo–maternal cross-talk in early pregnancy in sheep. 2024; Reproduction, 168(5), doi: 10.1530/REP-24-0172
• Dori P., Anastasio I., Macchi E., Manenti I., Hones M., Carosi M. Hibernating or not hibernating? Brown bears’ response to a mismatch between environmental natural cues and captive management, and its welfare implications. 2024; Plos one, 19(7), e0306537, doi: 1371/journal.pone.0306537
• Ozella L., Sartore S., Macchi E., Manenti I., Mioletti S., Miniscalco B., Crosetto R., Ponzio P., Fiorilla E., Mugnai C. Behaviour and welfare assessment of autochthonous slow-growing rabbits: The role of housing systems. 2024; Plos one, 19(7), e0307456, doi: 10.1371/journal.pone.0307456
• Bongiorno V., Gariglio M., Zambotto V., Cappone E. E., Biasato I., Renna M., Gasco L., Bergagna S., Manenti I., Macchi E., Gai F., Schiavone A. Organic medium‐growing chickens fed live black soldier fly larvae: A welfare improvement study. 2024; Journal of Animal Physiology and Animal Nutrition, 1–16, doi:10.1111/jpn.13997
• Miretti S., Manenti I., Toschi P., Macchi E., Martignani E., Accornero P., Baratta M. Bovine Skeletal Muscle Satellite Cells: Isolation, Growth, and Differentiation, 2024; Methods in Molecular Biology, 2749:165-174, doi:10.1007/978-1-0716-3609-1_15
• Toschi P., Viola I., Manenti I., Miretti S., Macchi E., Martignani E., Accornero P., Baratta M. Ovine Trophoblast Cells: Cell Isolation and Culturing from the Placenta at the Early Stage of Pregnancy, 2024; Epithelial Cell Culture. Methods in Molecular Biology, 2749:123-133, doi:10.1007/978-1-0716-3609-1_12
• Manenti I., Viola I., Ala U., Cornale P., Macchi E., Toschi P., Martignani E., Baratta M., Miretti S. Adaptation Response in Sheep: Ewes in Different Cortisol Clusters Reveal Changes in the Expression of Salivary miRNAs. 2023; Animals, 13(20): 3273, doi: 10.3390/ani13203273
• Viola I., Toschi P., Manenti I., Accornero P., Baratta M. Modulatory role of mTOR in trophoblast adaptive response in the early stage of placentation in sheep. 2023; Reproduction, 165(3):313-324, doi:10.1530/REP-22-0356
• Ogun S., Viola I., Obertino M., Manenti I., Ala U., Brugiapaglia A., Battaglini L., Perona G., Baratta M. Using sensors to detect individual responses of lambs during transport and pre-slaughter handling and their relationship with meat quality. 2022; Animal Welfare, 31(4), 505-516. doi:10.7120/09627286.31.4.010

Publications

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