Our data highlight the importance of mare microbiomes as a key factor for the establishment of the gut microbial ecosystem of the foal.

Both in human and equine species, mesenchymal stem cells (MSCs) from amniotic membrane (AM) and Wharton's jelly (WJ), may be particularly useful for immediate use or in later stages of life, after cryopreservation in cell bank. The aim of this study was to compare equine AM- and WJ-MSCs features that may be relevant for their clinical employment. MSCs were more easily isolated from WJ, even if MSCs derived from AM exhibited more rapid proliferation ( < 0.05). Osteogenic and chondrogenic differentiation were more prominent in MSCs derived from WJ. This is also suggested by the lower adhesion of AM cells, demonstrated by the greater volume of spheroids after hanging drop culture ( < 0.05). Data obtained by PCR confirmed the immunosuppressive function of AM and WJ-MSCs and the presence of active genes specific for anti-inflammatory and angiogenic factors (IL-6, IL 8, IL-β1). For the first time, by means of transmission electron microscopy (TEM), we ascertained that equine WJ-MSCs constitutively contain a very impressive number of large vesicular structures, scattered throughout the cytoplasm. Moreover, an abundant extracellular fibrillar matrix was located in the intercellular spaces among WJ-MSCs. Data recorded in this study reveal that MSCs from different fetal tissues have different characteristics that may drive their therapeutic use. These finding could be noteworthy for horses as well as for other mammalian species, including humans.

A complex feedback of growth factors, secreted by a variety of cell types, is responsible for the mediation of skin healing. Despite the recent advances in wound healing management, this fails up to 50% and skin wounds can still be considered one of the main causes of morbidity, both in human and veterinary medicine. Regenerative medicine, involving mesenchymal stromal cells (MSCs), is nowadays a promising solution for skin wound healing. Indeed, MSCs are involved in the modulation of the inflammatory local response and cell replacing, by a paracrine mode of action. Local application of equine umbilical cord Wharton's jelly MSCs (WJMSCS) was carried out in a 6-months-old filly with a non-healing skin wound. Heterologous WJMSCs were applied four times using a carboxymethylcellulose (CMC) gel, produced dissolving CMC in autologous plasma. At first application the mean wound area was 7.28 ± 0.2 cm2. Four days after the last application of WJMSCs, the mean wound area was 1.90 ± 0.03 cm2, and the wound regression rate was +74%. No local or systemic side effects were registered after WJMSCs application and no evident exuberant scar was observed after wound healing. At discharge, the mean wound area was 0.38 ± 0.01 cm2 and the total regression rate was +80%. Five days later, the wound was completely healed. In the present clinical case report, the use of WJMSCs led to promising clinical results, paving the way for possible future applications in the treatment of chronic wounds in horses.

In the present study, an innovative therapy for spontaneously arisen pressure sores in seven hospitalized neonatal foals is described, using mesenchymal stem cells obtained from equine amniotic fluid. Amniotic fluid mesenchymal stem cells (AFMSCs) were isolated from fluid samples recovered at delivery. Heterologous cells, at passage three of in vitro culture, were applied to sores twice a week for four consecutive times in a carboxymethylcellulose (CMC) gel. As a control, a commercial ointment was used. The results showed that the mean sore regression rate with AFMSCs in CMC gel was statistically higher than the mean value recorded in the control group. This was associated with a significant effect of the treatment used and a statistically significant effect of treatment over time. The results suggest that local application of mesenchymal stem cells isolated from amniotic fluid, using CMC as scaffold, could be considered as an effective treatment of deep sores in hospitalized neonatal foals.

Background Little is known about the differences among adult and foetal equine mesenchymal stem cells (MSCs), and no data exist about their comparative ultrastructural morphology. The aim of this study was to describe and compare characteristics, immune properties, and ultrastructural morphology of equine adult (bone marrow: BM, and adipose tissue: AT) and foetal adnexa derived (umbilical cord blood: UCB, and Wharton’s jelly: WJ) MSCs. Results No differences were observed in proliferation during the first 3 passages. While migration ability was similar among cells, foetal MSCs showed a higher adhesion ability, forming smaller spheroids after hanging drop culture ( P < 0.05). All MSCs differentiated toward adipogenic, chondrogenic and osteogenic lineages, only tenogenic differentiation was less evident for WJ-MSCs. Data obtained by PCR confirmed MHC1 expression and lack of MHC2 expression in all four cell types. Foetal adnexa MSCs were positive for genes specific for anti-inflammatory and angiogenic factors (IL6, IL8, ILβ1) and WJ-MSCs were the only positive for OCT4 pluripotency gene. At immunofluorescence all cells expressed typical mesenchymal markers (α-SMA, N-cadherin), except for BM-MSCs, which did not express N-cadherin. By transmission electron microscopy, it was observed that WJ-MSCs had a higher ( P < 0.05) number of microvesicles compared to adult MSCs, and UCB-MSCs showed more microvesicles than BM-MSCs ( P < 0.05). AT-MSCs had a lower number of mitochondria than WJ-MSCs ( P < 0.05), and mitochondrial area was higher for WJ-MSCs compared to UCB and AT-MSCs ( P < 0.05). Conclusions Results demonstrate that MSCs from adult and foetal tissues have different characteristics, and foetal MSCs, particularly WJ derived ones, seem to have some charactestics that warrant further investigation into potential advantages for clinical application.

Dystocia as a prolonged stage II parturition (>30 min) was associated with a higher risk of complications. The hypothesis of the study was that any type of dystocia could affect the foal’s health, even when the stage II was <30 min. Clinical reports on 222 Standardbred mares and their foals hospitalized at the Veterinary Teaching Hospital of the University of Bologna from 2004 to 2020 were reviewed. Mares were divided into the Eutocia Group (165, eutocic delivery) and the Dystocia Group (57, dystocic delivery). The incidence of dystocia was 4.9%. Stage II was longer in the Dystocia Group (median 20 min) than in the Eutocia Group (median 12 min). All occurrences of dystocia were retrospectively classified into three categories of severity: mild, moderate and severe dystocia. The occurrence of postpartum complications in mares and neonatal diseases and failure of passive transfer of immunity in foals was higher in the Dystocia Group. Foal venous lactatemia and serum creatine kinase were significantly higher in the Dystocia Group (median 3.9 mmol/L; 262 UI/L respectively) than in the Eutocia Group (median 3.1 mmol/L; 187 UI/L respectively). The APGAR score was lower in the Dystocia Group (median 8) than in the Eutocia Group (median 10) and significantly lower in severe dystocia (median 3). The duration of stage II should not be considered the only parameter of dystocia in mares: even a rapid resolution of dystocia could pose health risks to the foal and the mare.

Wharton's jelly (WJ) is an important source of mesenchymal stem cells (MSCs) both in human and other animals. The aim of this study was to compare human and equine WJMSCs. Human and equine WJMSCs were isolated and cultured using the same protocols and culture media. Cells were characterized by analysing morphology, growth rate, migration and adhesion capability, immunophenotype, differentiation potential and ultrastructure. Results showed that human and equine WJMSCs have similar ultrastructural details connected with intense synthetic and metabolic activity, but differ in growth, migration, adhesion capability and differentiation potential. In fact, at the scratch assay and transwell migration assay, the migration ability of human WJMSCs was higher (P < 0.05) than that of equine cells, while the volume of spheroids obtained after 48 h of culture in hanging drop was larger than the volume of equine ones (P < 0.05), demonstrating a lower cell adhesion ability. This can also revealed in the lower doubling time of equine cells (3.5 ± 2.4 days) as compared to human (6.5 ± 4.3 days) (P < 0.05), and subsequently in the higher number of cell doubling after 44 days of culture observed for the equine (20.3 ± 1.7) as compared to human cells (8.7 ± 2.4) (P < 0.05), and to the higher (P < 0.05) ability to form fibroblast colonies at P3. Even if in both species tri-lineage differentiation was achieved, equine cells showed an higher chondrogenic and osteogenic differentiation ability (P < 0.05). Our findings indicate that, although the ultrastructure demonstrated a staminal phenotype in human and equine WJMSCs, they showed different properties reflecting the different sources of MSCs.

In the mare, foaling is a critical unpredictable event due to a wide range of gestational length and the absence of clear signs of impending parturition. To predict foaling, pH, inversion sodium potassium and increase of calcium concentration in mammary secretions are used. The aim of this study was to find how many days are left until parturition knowing mare's age (A) and parity (P) combined with ultrasonographic measurements of the fetal orbit in Standardbred mares with normal pregnancy. Eighty healthy Standardbred mares with normal pregnancy were hospitalized for attended delivery. Information about mare's age, parity and breeding date were recorded at admission. Transrectal ultrasonography were routinely performed at admission and every 10 days until parturition using a B-mode real time portable unit equipped with a 5-7.5 MHz linear transducer. The images of the fetal orbit were acquired when cornea, anterior and posterior chamber, vitreous body, lens and optic nerve were visible. Longitudinal diameter (LD) was considered as the distance between the two ocular poles, within the vitreous body; transverse diameter (TD), perpendicular to LD and bisecting the lens, was measured as the distance between cornea and retina. At delivery, length of pregnancy and gestational age at each exam were registered. For each ultrasound examination, days before parturition (DBP) were calculated. Seventy-eight Standardbred mares with normal pregnancies were included in the study. Mares' mean age was 9 ± 5 years old (range 4-20 years) and mean gestation length was 341 ± 7 days (range 327-366 days). Thirty-three mares were primiparous and 45 mares were multiparous. Data were analyzed using a regression tree: P, A, LD and TD were used as covariates. DBP was used as the variable of interest. Nine terminal nodes were identified based on the selected covariates. The first split is produced by the TD: fetuses with TD greater or equal than 2.97 cm are further split according to LD, with a threshold of 3.28 cm. The next split is dictated by A; after a further split on LD, the first terminal node is built, containing 34 fetuses with average DBP equal to 10 days. If the A is ≥ 9.5 years a further split is on P: when mares are multiparous, the TD built two different nodes. Since prediction of mare's foaling date is an important factor in stud farm management, the regression model developed may help the veterinarian to estimate the DBP in Standardbred mares with normal pregnancy.