Background: In Piedmontese cattle the double-muscled phenotype is an inherited condition associated to a point mutation in the myostatin (MSTN) gene. The Piedmontese MSTN missense mutation G938A is translated to C313Y myostatin protein. This mutation alters MSTN function as a negative regulator of muscle growth, thereby inducing muscle hypertrophy. MiRNAs could play a role in skeletal muscle hypertrophy modulation by down-regulating gene expression. Results: After identifying a 3^′-UTR consensus sequence of several negative and positive modulator genes involved in the skeletal muscle hypertrophy pathway, such as IGF1, IGF1R, PPP3CA, NFATc1, MEF2C, GSK3b, TEAD1 and MSTN, we screened miRNAs matching to it. This analysis led to the identification of miR-27b, miR-132, miR-186 and miR-199b-5p as possible candidates. We collected samples of longissimus thoracis from twenty Piedmontese and twenty Friesian male bovines. In Piedmontese group miR-27b was up-regulated 7.4-fold (p < 0.05). Further, we report that the level of MSTN mRNA was about 5-fold lower in Piedmontese cattle vs Friesian cattle (p < 0.0001) and that less mature MSTN protein was detected in the Piedmontese one (p < 0.0001). Cotransfection of miR-27b and psi-check2 vector with the luciferase reporter gene linked to the bovine wild-type 3^′-UTR of MSTN strongly inhibited the luciferase activity (79%, p < 0.0001). Conclusions: These data demonstrate that bovine MSTN is a specific target of miR-27b and that miRNAs contribute to explain additive phenotypic hypertrophy in Piedmontese cattle selected for the MSTN gene mutation, possibly outlining a more precise genetic signature able to elucidate differences in muscle conformation.

Background Mesenchymal stem cells (MSCs) have been used therapeutically in equine medicine. MSCs release extracellular vesicles (EVs), which affect cell processes by inhibiting cell apoptosis and regulating inflammation. To date, little is known about equine EVs and their regenerative properties. Objectives To characterise equine MSC-derived extracellular vesicles (EVs) and evaluate their effect on equine chondrocytes treated with pro-inflammatory cytokines in vitro. Study design In vitro experiments with randomised complete block design. Methods Mesenchymal stem cells from bone marrow, adipose tissue, and synovial fluid were cultured in vitro. The MSC culture medium was centrifuged and filtered. Isolated particles were analysed for size and concentration (total number of particles per mL). Transmission electron microscopy analysis was performed to evaluate the morphology and CD9 expression of the particles. Chondrocytes from healthy equines were treated with the inflammatory cytokines interleukin (IL)-1β and tumour necrosis factor-alpha. MSC-derived EVs from bone marrow and synovial fluid cells were added as co-treatments in vitro. Gene expression analysis by real-time PCR was performed to evaluate the effects of EVs. Results The particles isolated from MSCs derived from different tissues did not differ significantly in size and concentration. The particles had a round-like shape and positively expressed CD9. EVs from bone marrow cells displayed reduced expression of metalloproteinase-13. Main limitations Sample size and characterisation of the content of EVs Conclusions EVs isolated from equine bone marrow MSCs reduced metalloproteinase 13 gene expression; this gene encodes an enzyme related to cartilage degradation in inflamed chondrocytes in vitro. EVs derived from MSCs can reduce inflammation and could potentially be used as an adjuvant treatment to improve tissue and cartilage repair in the articular pathologies.

The agricultural sector is amidst an industrial revolution driven by the integration of sensing, communication, and artificial intelligence (AI). Within this context, the internet of things (IoT) takes center stage, particularly in facilitating remote livestock monitoring. Challenges persist, particularly in effective field communication, adequate coverage, and long-range data transmission. This study focuses on employing LoRa communication for livestock monitoring in mountainous pastures in the north-western Alps in Italy. The empirical assessment tackles the complexity of predicting LoRa path loss attributed to diverse land-cover types, highlighting the subtle difficulty of gateway deployment to ensure reliable coverage in real-world scenarios. Moreover, the high expense of densely deploying end devices makes it difficult to fully analyze LoRa link behavior, hindering a complete understanding of networking coverage in mountainous environments. This study aims to elucidate the stability of LoRa link performance in spatial dimensions and ascertain the extent of reliable communication coverage achievable by gateways in mountainous environments. Additionally, an innovative deep learning approach was proposed to accurately estimate path loss across challenging terrains. Remote sensing contributes to land-cover recognition, while Bidirectional Long Short-Term Memory (Bi-LSTM) enhances the path loss model’s precision. Through rigorous implementation and comprehensive evaluation using collected experimental data, this deep learning approach significantly curtails estimation errors, outperforming established models. Our results demonstrate that our prediction model outperforms established models with a reduction in estimation error to less than 5 dB, marking a 2X improvement over state-of-the-art models. Overall, this study signifies a substantial advancement in IoT-driven livestock monitoring, presenting robust communication and precise path loss prediction in rugged landscapes.

Mesenchymal stem cells (MSCs) have been used in equines as an alternative therapy. A comparative study about the phenotype and in vitro performance of different MSCs tissue sources in adult equines was needed. This study might serve to provide the knowledge to select a valuable harvesting source of MSCs. Bone marrow, synovial and adipose (mesenteric, neck and tail fat) tissues were collected from adult equines. Cell surface markers expression (CD11α/CD18, CD45, CD79α, CD90, CD105 and MHC II) and in vitro differentiation assays were made. In vitro cell migration, cell growth and wound healing capacity tests helped to study their behavior and properties. MSCs phenotype was positively confirmed by the cell surfaces markers and a tri-lineage differentiation profile. Bone marrow cells showed the highest migration capacity, while synovial fluid cells displayed the highest cell growth. Bone marrow cells showed a better wound healing when compared with all the different MSCs. We conclude that bone marrow, synovial and adipose tissue derived from adult equines are a good source for cell therapy but they conserve different functional properties: bone marrow showed an interesting migration and wound healing capacity while synovial fluid cells and their highest cell growth suggest that these MSCs would yield higher cell numbers in a shorter time.

Mammary stem cells provide opportunities for the cyclic remodelling of the bovine mammary gland. Therefore, understanding the character and regulation of mammary stem cells is important for increasing animal health and productivity. The exciting possibility that stem cell expansion can influence milk production is currently being investigated by several researchers. In fact, appropriate regulation of mammary stem cells could hopefully benefit milk yield, persistency of lactation, dry period management and tissue repair. Accordingly, we and others have attempted to characterize and regulate the function of bovine mammary stem cells. However, research on mammary stem cells requires tissue biopsies, which represents a limitation for the management of animal welfare. Interestingly, different studies recently reported the identification of putative mammary stem cells in human breast milk. The possible identification of primitive cell types within cow's milk may provide a non-invasive source of relevant mammary cells for a wide range of applications. In this review, we have summarized the main achievements in this field for dairy cow science and described the interesting perspectives open to manipulate milk persistency during lactation and to cope with oxidative stress during the transition period by regulating mammary stem cells.