The aim of this study was to collect and analyze ultrasound measurements of fetal-maternal structures during normal and pathological pregnancies in jennies, a livestock species of growing interest. For two breeding seasons, 38 jennies of different breeds and crossbreeds aged between 3 and 18 years were monitored weekly by transrectal examination using a portable Esaote ultrasound (MyLab™ 30 GOLD VET) with a 5-7.5 MHz probe. The jennies were divided into two groups, < 250 kg and > 250 kg body weight, and the dates of conception and parturition/abortion were recorded to calculate pregnancy length. Descriptive statistics were performed for the following variables: pregnancy length and maternalfetal parameters (measurements of the orbit, gastric bubble, thorax, abdomen, gonads, heart rate, umbilical artery velocimetry, and combined utero-placental thickness). A total of 68 pregnancies were studied, 36 of which ended during the study period. The average pregnancy length was 370.82 ± 16.6 days for full-term pregnancies (N = 28, 77.8%) and 316.13 ± 36.6 days for abortions (N = 8, 22.2%). The season of conception and fetal gender did not affect the pregnancy length. Pregnancy examination can reasonably be performed by two weeks after last service if ovulation date is not known. The orbital diameter was the most reliable parameter for monitoring the physiological development of the embryo and fetus, and it was strongly related to the gestational age. No differences in fetal development were observed in relation to the mother's body weight. The combined utero-placental thickness was not associated with the gestational age and thickening and edema, frequently observed, were not associated with fetal pathologies.

Equine Papillomavirus 2 (EcPV2) is responsible for squamous cell carcinomas (eSCCs) of external genitalia of both male and female horses. However, few studies report the EcPV2 prevalence among healthy horses. Currently, the lack of these data does not permit identifying at-risk populations and, thus, developing screening protocols aimed at the early detection of the infection, as for humans. The aim of our study was to estimate the genoprevalence of EcPV2 in clinically healthy horses in Italy and to evaluate their innate immune response. For this purpose, penile and vulvar swabs of 234 healthy horses were collected through sampling with sterile cytobrushes. Nucleic acids were isolated and EcPV2-L1 presence (DNA) and gene expression (RNA) were checked by RT-qPCR. Our results showed EcPV2-L1 DNA presence in 30.3% of the samples and L1 expression in 48% of the positive samples. No statistically significant differences were found in genoprevalence in relation to sex, age, and origin, while, concerning breeds, the Thoroughbred had the highest risk of infection. Concerning specifically the mares, 40.2% of them resulted in being positive for EcPV2; our findings show a major positivity in pluriparous (p = 0.0111) and mares subjected to natural reproduction (p = 0.0037). Moreover, samples expressing L1 showed an increased expression of IL1B (p = 0.0139) and IL12p40 (p = 0.0133) and a decreased expression of RANKL (p = 0.0229) and TGFB (p = 0.0177). This finding suggests the presence of an effective immune response, which could explain the low incidence of SCCs in positive horses, despite a high EcPV2 genoprevalence (30%).

Papillomavirus (PV) infections may be related to anogenital lesions and cancer development in humans and several other animal species. To date, 11 different PVs have been reported in horses. Among them, a newly described PV named Equus caballus Papillomavirus Type9 (EcPV9) was thus far only reported in the semen of a stallion with penile lesions in Australia. This study reports for the first time the presence of EcPV9 in asymptomatic Italian horses. From July 2020 to January 2022, genital brush samples were collected from 209 horses with no apparent signs of neoplastic disease and no PV-associated lesions, clinically examined at the Didactic Veterinary University Hospital (OVUD) of Perugia and at the Veterinary University Hospital (OVU) of Turin. Brushes were submitted to real-time PCR targeting the EcPV9-L1 region. The first amplification targeted a region of ~116 bp, followed by the amplification and sequencing of ~533 bp of the positive samples. EcPV9-L1 DNA was found in eleven horses (5.3%), all female and mainly English Thoroughbred. Co-infection with EcPV2-L1 was found in 7 out of the 11 EcPV9-L1 positive horses (63.6%). This study contributes to the description of the prevalence of exposure or infection of EcPVs in the horse population in Italy, for which data are still limited. In this regard, here we provide a phylogenetic analysis and the completely reconstructed viral genomes of two Italian EcPV type 9 isolates, as well as four EcPV type 2 obtained from co-infected animals.

When endometritis becomes chronic in mares, infertility can follow. Among various causative agents, many bacteria are involved and mono-or mixed-infections are common. In our study, fifty mares with a previous history of subfertility were subjected to clinical and ultrasonographic examination of the reproductive tract, and samples were collected for cytology, bacteriology and PCR for Chlamydia spp detection. The aim of this work was to highlight the presence of Chlamydia abortus in chronic endometritis of subfertile mares. Endometrial chronic lesions were detected in five of six Chlamydia-positive animals.

The purpose of this study was to observe in vitro matured equine oocytes using a polarized light microscope (PLM) to compare the subjective morphological evaluation obtained using a classic light microscope (LM) with an objective computerized evaluation.Equine cumulus-oocyte complexes (COCs, n=922) were subjected to different in vitro maturation times (24, 36, or 45 h), but only 36-h matured oocytes were analyzed using PLM.The 36-h matured oocytes that reached maturity were parthenogenetically activated to evaluate quality and meiotic competence. Average maturation percentages per session in groups 1, 2, and 3 (24-, 36-and 45-h matured oocytes, respectively) were 29.31±13.85%, 47.01±9.90%, and 36.62±5.28%, whereas the average percentages of immature oocytes per session was 2 28.78±20.17%, 7.83±5.51%, and 22.36±8.39%, respectively. The zona pellucida (ZP) birefringent properties were estimated and correlated with activation outcome. ZP thickness and retardance of the inner layer of the zona pellucida (IL-ZP) were significantly increased in immature oocytes compared with mature oocytes (p<0.001 and p<0.01, respectively). The comparison between parthenogenetically activated and non-activated oocytes showed a significant increase in the area and thickness of the IL-ZP in parthenogenetically activated oocytes (p<0.01). These results show that the 36-h IVM protocol allowed equine oocytes to reach maturity, and PLM observation of ZP can be used to distinguish mature and immature oocytes as well as activated and non-activated oocytes.

No abstract

The selection of the recipient mare is one of the most important factors involved in the success of equine embryo transfer. The aim of this study was to determine whether the age of the recipient, the number of transfers and the detection of intrauterine fluid during the follicular phase or after ovulation can affect pregnancy rate at 14 and 45 days (PR 14 and PR 45) or early embryonic loss (EEL). A total of 1222 ETs were included in the study. Mares receiving the first embryo of the year had a higher PR 14 and 45 days compared to mares at the third transfer (78.8% and 70.1% vs. 65.6% and 54.1%, respectively). The detection of intrauterine fluid post ovulation negatively affected PR 14 (60.5% vs. 77.6%) and should therefore be considered an abnormal finding, probably being a sign of uterine inflammation or delayed uterine clearance. On the contrary, the age of the recipient mare and detection of fluid during follicular phase did not affect PR 14. Only the age of the recipient mare influenced the EEL, since mares aged 10–13 years had a higher EEL compared to mares aged 3–5 years (15.6% vs. 6.4%). Embryo size and grade affected PR 14 and 45.

Des inséminations artificielles sur chaleurs induites et chaleurs naturelles ont été effectuées sur 138 femelles zébus Azaouak à la Station sahélienne expérimentale de Toukounous. Les chaleurs ont été induites par l’utilisation de l’implant Crestar, de la spirale vaginale, et d’injections de prostaglandine et de gonadolibérine (GnRH). Deux inséminations ont été faites : la première à l’observation des chaleurs et la seconde 12 heures après. L’influence de paramètres zootechniques a été testée. Les taux non cumulés de gravidité obtenus ont été de 24,5 p. 100 à la première insémination, 33,72 p. 100 à la deuxième, et 29,48 p. 100 à la troisième et au-delà. L’utilisation de semence fraîche, et l’utilisation de vaches et non de génisses ont fait partie des paramètres zootechniques influençant significativement et positivement les inséminations. Le délai de 60 jours post partum généralement observé avant les inséminations n’a pas eu d’effet significatif sur les résultats. La saison d’insémination n’ayant pas influencé les résultats, des mises bas ont pu être observées tout au long de l’année. L’état nutritionnel a par ailleurs été un facteur essentiel de réussite des inséminations ; les résultats ont traduit la capacité des zébus Azaouak à valoriser les fourrages en toute saison, avec une reprise rapide de l’activité ovarienne après la mise bas.