Maria Augusta Alonso

Addition of seminal plasma to post-thawing equine semen: what is the effect on sperm cell viability?

Effect of seminal plasma addition after thawing on viability or cryocapacitation is not definitively established. This experiment was performed to verify the effect of adding seminal plasma, autologous or homologous (from an animal with good semen freezability). Five ejaculates from each of four stallions with proven fertility were collected and cryopreserved. The semen was subsequently thawed and divided into the following three treatment groups: no seminal plasma addition after semen thawing (NOSP); the addition of homologous seminal plasma after semen thawing (HSP) and the addition of autologous seminal plasma after semen thawing (ASP). The addition of 20% of seminal plasma led to an increase in the cell population that simultaneously show plasma and acrosomal membrane integrity (p < 0.05). The addition of seminal plasma did not alter the total motility, the amount of cells with mitochondrial membrane potential or the sperm velocities (average path velocity, straight-line velocity and curvilinear velocity). However, the beat/cross-frequency, straightness and linearity were reduced in ASP and HSP groups (p < 0.05). Unexpectedly, the addition of homologous seminal plasma reduced the proportion of cells with progressive motility (p < 0.05) and the addition of autologous seminal plasma reduced the amplitude of the lateral head displacement (p < 0.05). Based on the increase in the cell populations that had the plasma and acrosomal membrane integrity simultaneously identified in this study, we proposed that the addition of seminal plasma (autologous or homologous) into post-thawed semen before insemination could increase semen fertility.

Nutraceuticals in reproduction of bulls and stallions

The industry has made available in the market a series of substances (nutraceuticals) which intent would be to optimize the use of nutrients in some metabolic paths, influencing positively reproductive performance in animals. However, the response to the use of nutraceuticals varies according to the animal. As the organism is highly complex and in order to achieve a perfect activity of the hypothalamic-pituitary-gonadal axis, an ideal interaction in molecular basis is needed, where the nutraceuticals can have their direct action. The aim of this study was to review the function and research results using the main nutraceuticals (β carotene, vitamin A, L-carnitine, omegas 3, 6 and 9 and Gamma-oryzanol) on reproductive characteristics of bulls and stallions.

Influence of Maternal Age and Parity on Placental Structure and Foal Characteristics From Birth up to 2 Years of Age

ABSTRACT The foals weight and height at birth are regarded as important parameters for the equestrian sports industry. Moreover, scientific studies indicate that maternal environment can influence postnatal genetic potential of foals. The purpose of this study was to evaluate the influence of maternal age and parity on placental efficiency and size of the newborn foal using stereological analysis tools of the terms allantochorion. Furthermore, the influence of maternal age and parity on weight, height, and girth circumference of the offspring from birth until 2 years of age was investigated. Forty pregnant mares were categorized according to age: 4–8 years (n = 15); 9–12 years (n = 15); ≥13 years (n = 10) and parity: nulliparous (n = 12), 1–3 parities (n = 18); ≥4 parities (n = 10). Parturition was monitored and the allantochorion membranes were collected after delivery. The stereological analysis determined the volumetric composition of the different compartments of the placenta along with the area of fetomaternal contact surface. It was observed that maternal age and parity significantly increased the development of the microcotyledons and total villi surface density, as well as the vascularization and tissue composition of different regions of the placenta, which contributes to fetomaternal contact. We also found a significant relationship between maternal age and parity on the size of the neonate foals but not after the first year of age, possibly because environmental and genetic factors contribute to postnatal growth. HIGHLIGHTSMaternal age and parity influence placental characteristics and fetomaternal contact.Mares age and parity influence neonatal size at birth.After the first year, differences in height, weight, and girth circumference disappear.

Uterine Vascular Perfusion and Involution During the Postpartum Period in Mares

ABSTRACT In horses, limited data is found regarding the vascular events during uterine involution at the puerperal period. Thus, we aimed to evaluate the morphological aspects (size of uterus and intrauterine fluid content) and the hemodynamics (endometrial and mesometrial vascular perfusion) of the uterus during its postpartum involution process. Ten mares were daily scanned by transrectal color Doppler ultrasonography from the first day postpartum (d1) to the 16th day after first postpartum ovulation (D0 = ovulation). The formerly gravid horn (GH) and formerly nongravid horn (NH) were individually evaluated. A reduction (P < .05) in the uterine diameter was observed during the first 7 days postpartum, and the rate of uterine involution decreased after this period. The involution was completed on d21 and d24 for the NH and GH, respectively. Presence of intrauterine fluid was present in large amounts between d1 and d2 postpartum, followed by a decrease (P < .05) between d4 and d7. No fluid was observed after d16 postpartum or after the third day postovulation (D3). During the early postpartum period, an increase (P < .05) in the endometrial and mesometrial vascularization was detected, respectively, between d1 and d4, and between d1 and d2. The vascular perfusion did not differ after d4 for endometrial tissue, whereas was reduced (P < .05) between d2 and d10 for mesometrium. After the first postpartum ovulation, an increase (P < .05) in vascular perfusion was observed from D0 to D5, followed by a decrease (P < .05) between D5 and D11 and an increase (P < .05) between D11 and D14. The novel vascular perfusion profile here described in the endometrium and mesometrium after ovulation is similar to the uterine vascular profile observed during estrous cycles and early pregnancy, indicating a fast return of the mares uterus to cycling postpartum conditions. HIGHLIGHTSImportant physiological changes involved in the uterine involution were described.Great reduction in the uterine diameter was observed during the first days postpartum.The uterine involution process was completed until 4 weeks postpartum.Uterine vascularity has a transitional increase during first 2 to 4 days postpartum.Uterine vascularity after ovulation is similar to that observed during early pregnancy.

Factors affecting gestational length in the Mangalarga Paulista breed

The Mangalarga Paulista is a Brazilian horse breed that originated from a cross between Andalusian and Alter Real horses, and despite its national importance, no data exist regarding their gestational length. This study was conducted to determine the variation in gestation length and the influence of factors such as the sex of the foal, maternal age and parity, stallion and mating period. Retrospectively, data from 208 gestations of Mangalarga Paulista foals located in Piracaia, SP, Brazil, during the period from 1991 to 2012 were analyzed. The mean gestational length was 343.08 ± 0.66 days, with a range between 317 and 404 days. Gestation of mares carrying colts had a tendency to be longer than mares carrying fillies. Maternal parity significantly affected gestational length, with longer gestation for primiparous mares until the third gestation, shorter pregnancies between 4th and 8th gestation, and longer thereafter. Regarding mating period, mares that were mated at the end of the breeding season (January and February) had shorter pregnancies (P < 0.05). These results are of significant clinical and economic relevance for the equine industry, so that the monitoring of gestation and parturition can be performed effectively.

Transplacental transmission of Theileria equi in mules: Should we worry?

Equine piroplasmosis, disease globally responsible for significant economic losses, is caused by the transmission of the hemoprotozoa Theileria equi and Babesia caballi by ticks. Transplacental transmission has been reported previously in horses, which is not the case for mules´ foals. The present study investigated, by Real Time-PCR (RT-PCR), the presence of both Theileria equi and Babesia caballi in newborn mules before the ingestion of colostrum. Blood samples of 18 mares and their respective mules´ foals were collected and analyzed by RT-PCR. Only one mare was positive for B. caballi. In contrast, 13 mares were positive for T. equi, with two foals (11.1%) positive for the same protozoon, both of affected mares. We may conclude that transplacental transmission of Theileria equi also occurs in mules which may be an important concern while breeding such animals.

Carnosine as malondialdehyde scavenger in stallion seminal plasma and its role in sperm function and oxidative status

Semen biotechniques may impair sperm quality due to excessive production of reactive oxygen species (ROS). Additionally, products of the oxidative reaction, especially involving lipids (e.g., malondialdehyde - MDA), may be even more harmful to sperm. Carnosine, previously reported to be present in seminal plasma of several species, may be a key factor on sperm tolerance to biotechniques by counterattacking the deleterious influence of MDA. Therefore, the aim of this study was to measure the levels of carnosine present in equine seminal plasma and relate these findings with sperm function and oxidative status during cooling and cryopreservation. Thus, semen samples were collected from 40 stallions in duplicate (N = 80) and then submitted to cooling and cryopreservation. Samples were then allocated into groups of high and low tolerance to refrigeration and cryopreservation (bad cooler and good cooler/bad freezer and good freezer, respectively), and in groups of different concentrations of carnosine (High, Medium-high, Medium-low and Low carnosine). Samples were evaluated for sperm kinetics patterns, function of sperm structures and oxidative status. In good cooler samples, it was observed higher concentrations of carnosine (Good cooler: 224.98 ± 19.16 ng/mL; Bad cooler: 159.72 ± 15.99 ng/mL; p = 0.0056), ROS production (Good cooler: 26.40 ± 18.33%; Bad cooler: 18.33 ± 1.84%; p = 0.001) and lipid peroxidation rates (Good cooler: 193.23 ± 18.22 ng/mL; Bad cooler: 131.92 ± 12.25; p = 0.0064). Groups of samples with higher carnosine concentrations had lower levels of malondialdehyde (High: 79.33 ± 6.72 ng/mL; Medium-high: 140.45 ± 11.70 ng/mL; Medium-low: 202.57 ± 16.30 ng/mL and Low: 231.02 ± 32.35 ng/mL; p < 0.05), demonstrating that carnosine was effective in removing lipid peroxidation products. Due to the removal of seminal plasma during the cryopreservation process, no differences occurred in carnosine levels between bad and good freezer groups. In this context, this study provides relevant data for future therapies using carnosine during cryopreservation, aiming to replace the levels lost due to the necessary removal of seminal plasma.