Eleonora Iacono

97 PRELIMINARY DESCRIPTIVE STUDY OF EQUINE PLACENTA GENERATED AFTER TRANSFER OF IN VIVO- AND IN VITRO-PRODUCED EMBRYOS

Placental changes associated with artificial reproductive technologies have been described in several species, but little information is available in horses. Joy et al. (2012) reported that human placentas from intracytoplasmic sperm injection derived embryos were heavier and thicker than those produced after natural conception. Despite the most growing interest and efficiency of artificial reproductive technologies in equine species, only recently, Pozor et al. (2016) described placental abnormalities in pregnancies generated by somatic cell NT, but there are no studies on equine placenta generated by intracytoplasmic sperm injection and traditional embryo transfer. In the present preliminary study, macroscopic differences of placentas generated after transfer of in vitro- or in vivo-produced embryos were registered. Twelve Standardbred recipient mares with pregnancy generated after transfer of in vivo-derived (Group 1) and in vitro-derived (Group 2) embryos were enrolled; 10 Standardbred mares with pregnancy derived by traditional AI were included as control (Group 3). All pregnancies were physiological, and newborn foals were healthy. Mare age, parity, length of pregnancy, gross evaluation and weight of placenta, total length of umbilical cord (UC), length of UC, number of UC coils, foal sex, and weight at birth were registered. Collected data are listed in Table 1 and are expressed as mean±standard deviation. Differences between groups were evaluated by 1-way ANOVA, and the difference in proportion of overweight placentas was evaluated with the Fisher test. The gross evaluation of placenta revealed 8/12 placentas (2/4 Group 1; 6/8 Group 2) were heavier than 11% (Madigan, 1997) due to oedema of the chorioallantois. No overweight placentas were registered in Group 3. In Group 1, 1/4 placentas had villous hypoplasia, and in Group 2, 1/8 placentas had cystic pouches on the UC. There were no significant differences among groups. However, the proportion of overweight placentas between Group 2 (6/8) and Group 3 (0/10) approached significance (P=0.06). Although preliminary, the results of the present study suggest that production of equine embryos in vitro may lead to alterations in placental development. Several studies in cattle and sheep have suggested that alterations in the placentas of pregnancies derived from in vitro-produced embryos are related to effects of culture on epigenetic regulation. Less is known in the horse about the effects of in vitro embryo production on placental development; thus, further research in this area is necessary.

Detection and quantification of Cryptosporidium oocysts in environmental surfaces of an Equine Perinatology Unit

The presence of Cryptosporidium in institutions such as veterinary teaching hospitals, where students and staff are in frequent contact with animals, could represent a serious public health risk. In this study the detection and quantification of the Cryptosporidium oocysts present on the environmental surfaces of an Equine Perinatology Unit (EPU) were investigated. During 3 foaling seasons 175 samples obtained by swabbing an area of the floor and walls of boxes and utility rooms of EPU with sterile gauze, in 3 different moments. Samples were collected at the end of foaling season (July), after washing procedures (September) and after washing and disinfecting procedures, at the beginning of a new foaling season (December). All the samples were subjected to nested-PCR, followed by genotyping and sub-typing methods and to qPCR, allowing the oocyst quantification. Cryptosporidium spp. was detected in 14 samples, of which 11 were from walls and three were from floors. The highest number of oocysts was found in a sample collected from the floor of one utility room used for setting up therapies and treatments. In most cases, oocyst numbers, estimated by qPCR, were reduced or eliminated after washing and disinfecting procedures. The genotyping and sub-typing methods allowed identification of 2 subtypes of C. parvum (IIaA15G2R1 and IIdA23G1) and 1 of Cryptosporidium horse genotype (VIaA15G4) that were described in foals hospitalized at the EPU in the same years. The results of the present study show that qPCR can be used to evaluate Cryptosporidium contamination of environmental surfaces of a veterinary teaching hospital and the efficacy of the disinfection procedures.

Effects of induced endometritis on uterine blood flow in cows as evaluated by transrectal Doppler sonography

This study was conducted to evaluate the effects of induced endometritis on uterine blood flow in cows. Transrectal Doppler sonography was performed on uterine arteries of six cyclic cows before and for 4 days after inducing acute endometritis by intrauterine infusion of 720 mg of policresulen, and for 4 days of the following estrous cycle. Time-averaged maximum velocity (TAMV) increased (p < 0.001) and pulsatility index (PI) decreased (p < 0.0001) within 1 h of policresulen administration, and did not change (p > 0.05) in the next 4 days of the same cycle. TAMV and PI values in the subsequent cycle did not differ (p > 0.05) from the values measured before infusion and showed no changes (p > 0.05) within the cycle. Blood flow parameters were not related (p > 0.05) to plasma concentrations of progesterone and estrogen. All cows showed an acute endometritis determined by histopathological findings of biopsy samples taken 1 day after infusion and fibrotic endometrial alterations detected in the subsequent cycle. No relationships were observed between fibrotic changes of the endometrium and uterine blood flow during either cycle. In conclusion, acute inflammation is accompanied by a rise in uterine blood flow, but fibrotic alterations do not seem to be related to Doppler sonographic findings.

Comparative characterization of human and equine Wharton’s jelly derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) have the capability to differentiate into wide range of specialized cells of mesodermal origin such as osteocytes, chondrocytes, adipocytes, cardiomyocytes, muscle fibers. Due to these properties, MSCs are considered as a new emerging treatment option and therapeutic agent in regenerative medicine. Promising results have been obtained after application of MSCs for treating tendon and joint disease in the equine model, making it favorable for therapeutic application. While the horse is considered a highly suitable model for orthopedic diseases, knowledge is lacking regarding the level of analogy of equine MSCs and their human counterparts. Therefore, the aim of this study was to assess the properties of human and equine Wharton’s jelly derived MSCs in a direct comparison. Obtained MSCs, were characterized for their staminal markers, proliferation and adhesion potential, ultrastructural morphology and their ability in differentiate towards osteogenic, chondrogenic and adipogenic lineages. Results showed a similar pattern in the expression of staminal markers, while a light difference was observed in the proliferation and adhesion potential. Ultramorphological analysis showed nuclear and citoplasmatic features comparable in human and equine MSCs. Finally, both MSCs were able to differentiate towards osteogenic, chondrogenic and adipogenic lineages. In conclusion, although revealing some potentially relevant differences, the study demonstrates a high level of analogy between human and equine MSCs, providing a basis for translational research in the equine model.