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Supplementary Files: Development of a Composite Pain Scale in Foals: A Pilot Study

The purpose of this PhD thesis is to perform a dissertation about the equine umbilical cord (UC), with particular attention to its macroscopic characteristics moving into its microscopic features. Furthermore, equine UC was microscopically and immunohistochemically described. The coiling makes the UC a structure both flexible and strong, and provides resistance to external forces, which could compromise blood flow. Data obtained from the present study could be considered as a normal range of number of coils and UCI in the equine physiological pregnancies. The second and major component of this dissertation was focused on umbilical cord as a Mesenchymal Stem Cells (MSCs) niche. In particular, the ultrastructural characterization and immune profile of equine MSCs derived from foetal tissues (Wharton’s jelly and blood) were investigated; equine foetal MSCs were then compared to human foetal MSCs and equine adult MSCs. The third section included the clinical applications of MSCs derived from AF and WJ on cutaneous wounds of different ages’ foals. The employ of foetal MSCs on cutaneous wounds decreased their healing both in neonatal foals and six-month foals, in comparison with other traditional treatments. In conclusion, one of the most important feature of placenta’s evaluation is the observation of normal characteristic of the equine UC. Furthermore, since the UC is a discarded material after delivery, it represents an excellent non-invasive alternative source of MSCs with reliable migration and differentiation capacities, and it is a convenient cell source for autologous or allogeneic regenerative therapies. Finally, it is important a constant update about regenerative medicine and the development of research in this field considering that the horse serves as a perfect model for humans.

none 5 no In foals more than 12 hours old, the only effective therapy for the treatment of failure of passive transfer (FPT) of immunity is transfusion of equine plasma. Use and efficacy of equine plasma for prophylaxis and treatment of sepsis, a condition primarily associated with FPT, are widely reported. However, plasma- and recipient-related factors associated with extent of IgG transfer and catabolism are not completely defined. Efficacy and safety of transfusion of a commercial fresh-frozen hyperimmune plasma were evaluated in hospitalized foals younger than 7 days of age with total or partial FPT. Sixty-two foals, classified as affected by FPT only, septic (infection plus systemic inflammatory response syndrome [SIRS]), and nonseptic sick, were included, and serum IgG concentration was measured at admission and 24 hours after plasma transfusion. In 25/62 foals, IgG level after 72 hours was also determined. The impact of different classification criteria for septic foals on IgG transfer was evaluated. Serum IgG measured 24 hours and 72 hours after plasma transfusion was significantly greater than at admission, but no significant difference was found in transfer efficacy (TE) between FPT, FPT septic, and FPT nonseptic foals and no significant difference was found in IgG concentration comparing foals with total and partial FPT or survivors and nonsurvivors. No significant difference was found comparing IgG concentration between bacteremic and nonbacteremic foals and foals with or without SIRS. No foal experienced adverse reactions to plasma transfusion. IgG TE and catabolism did not result significantly affected by the presence of sepsis or illness or by the outcome. none Freccero F., Mariella J., Lanci A., Cotignoli C., Castagnetti C. Freccero, F.; Mariella, J.; Lanci, A.; Cotignoli, C.; Castagnetti, C.

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.

Objectives The umbilical cord (UC) plays an essential role in the exchange between the mother and the foetus, allowing the latter to receive oxygen and nutrients necessary for survival and development. The UC intervascular stroma consists of mucous connective tissue, also called Wharton's jelly (WJ). It develops from extraembryonic mesoderm, binds and encases the umbilical vessels, protecting them from twisting and compression during pregnancy. WJ also has angiogenic and metabolic roles for the umbilical circulation. Some authors demonstrated that in human, an alteration of its composition could be related to fetal pathologic condition (Ferguson et al 2009). The aims of the study were to macroscopically describe the equine UC, and, for the first time in the horse, to evaluate the weight and the microscopic composition of WJ. Materials and Methods Twenty-four mares with normal pregnancy and parturition have been enrolled. The following data were registered for each mare: age, parity, length of pregnancy, total length of UC, length of UC amniotic and allantoic portion, total number of UC coils and their number in each segment, sex and weight of the foal at birth. There was no control of mare breeding. The Kolmogorov-Smirnov test was used to evaluate the distribution of continuous variables; the presence of differences in the UC macroscopic structure between gender was evaluated by T Student, considering significant a P value

Aim of the study: The purpose of this study was to evaluate the efficacy and safety of the administration of a commercial hyperimmune plasma in septic and non-septic foals with failure of passive transfer (FPT). Materials and methods: Fifty-two hospitalized foals

Total cholesterol (TC), HDL- and LDL-cholesterol have never been investigated in neonatal foals. The aims of this study were: a) to measure serum TC, HDL- and LDL-cholesterol in healthy foals at birth (T0) and after 72 hours (T72), and in septic foals at admission, b) to evaluate if any difference exists between healthy and septic foals and between surviving and non-surviving septic foals. Twenty-five foals ≤3 day-old were included: 13 healthy and 12 septic foals (mean age 24 ±18 hours), on the basis of positive blood culture and systemic inflammatory response. TC, HDL- and LDL-cholesterol were measured with colorimetric methods. Data were analyzed with paired and independent t-test, or 1-way ANOVA, and reported as mean ±sd. In healthy foals, TC, HDL- and LDL-cholesterol at T0 (TC 143 ±28 mg/dL; HDL- 31 ±12 mg/dL; LDL-cholesterol 101 ±27 mg/dL) were significantly lower than at T72 (TC 185 ±49 mg/dL; HDL- 50 ±6 mg/dL; LDL-cholesterol 124 ±43 mg/dL) (p

This study aimed to determine alpha-fetoprotein (AFP) concentrations in amniotic fluid, plasma of mares and respective foals: carrying normal pregnancies and delivering healthy foals (n=20; Group 1); carrying apparently normal pregnancies and delivering sick foals (n=15; Group 2); carrying high-risk pregnancies and delivering sick foals (n=14; Group 3). High-risk pregnancy was defined by a history of premature udder development/lactation or increased of the combined thickness of the uterus and placenta, or vulvar discharge and/or mares' systemic illness. Sick foals were affected by neonatal encephalopathy, sepsis, prematurity/dysmaturity, or hypoxic-ischemic encephalopathy. Based on histological examination of the chorioallantois, AFP trend was analyzed in pregnancies with pathologic (PFM) and normal fetal membranes (NFM). Concentrations of AFP were measured using a commercially available immunoassay previously validated for horses. Mares' plasma AFP did not change during the last 15-20 days of pregnancy in the three groups, and there was no difference among them. Amniotic fluid AFP was higher in Group 3 (P=.014). Foals' plasma AFP concentration was higher from birth to 72hours in foals of Group 2 and 3 than in healthy ones, and foals of Group 3 had the highest value. The strong association (r=0.84; P < .0001) between AFP in amniotic fluid and foals' plasma at birth is likely due to the presence of AFP in fetal urine. AFP was higher in pregnancy with PFM than with NFM in mare's plasma at admission (P=.031), amniotic fluid (P=.004), foal's plasma at birth (P=.002), at 24 (P=.005) and at 72 hours of life (P=.004). AFP is higher in pregnancy with histopathological lesions of the chorioallantois providing the evidence of the differences between pregnancy with a normal placental barrier and the more compromised ones. The increased AFP concentration in the amniotic fluid and plasma of high-risk foals suggests upregulation.