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Two studies were conducted to evaluate the use of medical ozone (O 3 ) in commercial extenders for equine semen cryopreservation. In the first study (Study 1), 0, 5, and 15 µg/mL of O 3 were added to diluents of refrigerated or frozen semen. Samples were evaluated for sperm kinematics at different time points for the chilled samples and after a thermoresistence test for the frozen/thawed samples. In the second study (Study 2), 0, 5, and 10 µg/mL of O 3 were added to an antibiotic-free diluent for refrigerated semen for comparison with the control group in which semen was diluted in the same diluent enriched with antibiotics. Semen sample kinematics were analyzed and an aliquot was collected after ozonification for bacteriological analyses. For Study 1 no difference was found comparing all the kinematic parameters analyzed over time, in the various treatments ( P > 0.05). In Study 2 the absence of antibiotics did not affect the kinematic parameters compared to the control ( P > 0.05). However when antibiotics were added, a smaller number of bacterial colony-forming units were detected compared to samples without antibiotics and without or with different O 3 supplementations. In conclusion, O 3 treatment at low dosages did not affect the semen kinematics, although it was ineffective in preventing bacterial overgrowth. Higher O 3 concentrations should be evaluated to explore the possibility of reducing the use of antibiotics in equine sperm conservation., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors. Published by Elsevier Ltd.)

Despite their inability to reproduce naturally, mules can host embryos and be surrogate dams. The aim of this investigation was to increase our knowledge of the qualitative composition of mule's milk and its variations throughout the whole lactation period-namely, from 6 h after foaling to 180 days in milk (DIM). Milk was obtained from a mule dam that had foaled after receiving a mule embryo transfer. For each sample, the gross, mineral, and fatty acid composition was evaluated. The average quality of the mule milk was as follows: protein 1.97 g 100 mL -1 , fat 0.90 g 100 mL -1 , and ash 0.39 g 100 mL -1 . Saturated fatty acids made up, on average, 50.00 g 100 g -1 of fat. Monounsaturated and polyunsaturated fatty acids made up half of the total fatty acid content (31.80 g 100 g -1 and 18.2 g 100 g -1 of fat, respectively). Linoleic acid and linolenic acid were the main polyunsaturated fatty acids in the milk. The milk composition changed throughout lactation. Dry matter, protein, fat, and ash decreased significantly from early lactation (6 h to 14 DIM). The n3 polyunsaturated fatty acids decreased at the end of lactation. The changes in milk composition throughout lactation are probably due to adaptations to the growth requirements of the foal.

Horses and donkeys differ phenotypically and karyotypically, although they can interbreed freely. Eight Standardbred mares and nine Amiata donkey jennies were included in the study. Semen was collected from two horses and two donkey stallions of proven fertility. A first pregnancy diagnosis was performed on day 10 after ovulation and repeated every day until embryo detection or until day 16. Irrespectively of the sire species, pregnancy rates in horse females (20/30, 66.7%) were significantly higher than in donkey females (19/70, 27.1%) ( p < 0.05), while horse and donkey males did not affect pregnancy rates. Comparing overall intraspecific and interspecific AI, pregnancy rates were 25/37 (67.6%) and 14/63 (22.2%), respectively ( p = 0.0001). The lowest pregnancy rate was obtained when inseminating jennies with horse stallion semen (8/49, 16.3%). No statistical differences were found when comparing embryo diameters, day at first pregnancy diagnosis, or in vitro embryo morphological quality among groups. In this study, much poorer results were obtained with jennies than with mares. Interspecific AI resulted in lower pregnancy rates than intraspecific Al, and AI to produce hinny hybrids resulted in the lowest pregnancy rate. Further studies are required to better understand the mechanism involved in such different outcomes in relation to intra- and interspecific breeding in domestic equids.

Due to the long courtship needed to attain excitation and erection, donkey semen collection can take up to 90 min. ProstaglandinF2α (PGF2α) has been reported to hasten the onset of erection and ejaculation in domesticated mammals, presumably by inducing smooth muscle contractions in the internal genitalia. However, while it has been anecdotally used in donkeys, it has yet to be critically evaluated. This study aimed to compare behavioral and semen parameters in Catalan, Balearic, Amiata, and Miranda jacks treated with the PGF2α analogue cloprostenol sodium immediately prior to exposure to an estrus jenny. Nineteen donkeys were assigned in a crossover design to receive cloprostenol sodium (125µg, i.m.; n = 53 collections) or saline (1 mL, i.m.; n = 53 collections). There were no differences for erection (52/53 vs. 52/53) or ejaculation (52/53 vs. 48/53) for collection attempts assigned to saline or cloprostenol sodium, respectively. Cloprostenol sodium significantly hastened treatment-to-erection and treatment-to-ejaculation times from 12.0 ± 1.6 to 6.0 ± 1.6 min and from 14.0 ± 1.4 to 9.6 ± 1.4 min, respectively. Significant effects of breed and age were observed in behavioral and parameters, but there were no effects of cloprostenol sodium administration on semen parameters. In conclusion, cloprostenol sodium administration immediately prior to semen collection hastened time to collect semen in donkeys with no detrimental effects on semen quality and can be used by practitioners to circumvent long delays in donkey semen collection.

Immunoglobulin type G (IgG) concentration both in jennies' colostrum and in serum of donkey foals are mostly unknown in the first 24 h after delivery. The aims of the present study were to evaluate the IgG concentrations of colostrum during the first 24 h of lactation of Amiata jennies, the absorption of colostrum and the weekly body weight gain of the donkey foals. IgG concentrations were assessed in the jennies' colostrum and in the serum of donkey foals. Colostrum was collected in 9 jennies ready after delivery, and at 6, 12, 24 h after foaling from both halves. Serum was collected at the same sampling times from 9 donkey foals. Donkey foals were weighted at birth and then weekly until the 28 th days of life. Temporal changes of IgG concentrations in dam's colostrum and in donkey foal serum were analyzed by a linear regression model and a general linear model, respectively. Results showed that colostrum IgG concentration were similar between the left and the right half. Colostrum IgG concentrations decreased continuously throughout the time in all jennies by 0.0244 Log 10 mg/mL per hour. Serum IgG concentrations in donkey foals at birth was significantly lower compared to other times. No correlation was found between the colostrum IgG concentrations and the average weekly body weight gain of the donkey foal. The pattern of colostrum IgG levels in jennies and serum IgG concentration in donkey foals seem to be similar to what reported for equine. However, the donkey foals seem to be less agammaglobulinemic at birth compared to the horse foal. The pattern and both serum and colostrum concentrations evaluated in the Amiata donkeys were slightly different from results reported in other donkey breeds, underlying the importance of setting references specific to breed., (© 2020 The Author(s).)

Obesity has been identified as a risk factor for developing breast cancer in post-menopausal period in humans and has been suspected to be associated with a worse prognosis also in the bitch. The aims of this study were to investigate the association between body condition score (BCS) and the prognosis of canine mammary carcinomas (CMCs) and the relationships between adiponectin expression and tumour behaviour. Seventy-three bitches with tubular, tubulopapillary, solid or complex carcinomas were included in the present study. For each dog, evaluation of BCS was conducted using a nine-point BCS system and the study population was divided into normal weight (4-5/9 points; n = 42), overweight (6-7/9 points; n = 19) and obese (8-9/9 points; n = 12). Type of diet (commercial, homemade or mixed) was recorded. After surgical excision, histological type, tumour size and nodal status were assessed and adiponectin expression was determined and quantified by immunohistochemistry and morphometric analysis. CMC histotype was not correlated with BCS, while a positive correlation between BCS and histological grade (p < .01) was observed. Overweight and obese bitches combined showed a shorter cancer-specific survival than normal weighted bitches (p < .01). Bitches fed with a homemade diet had a higher BCS than dogs fed with a commercial one, although no relationship was observed between diet and  cancer-specific survival. Thirty-six CMCs scored positive for adiponectin expression (49%), but no correlation was found between the hormone expression and either CMC characteristics or prognosis. In conclusion, a higher BCS seems to be related with a higher prevalence of more aggressive CMCs and negatively affects the survival time in bitches with these mammary tumours., (© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)