Melatonin reduces apoptotic cells,SOD2 and HSPB1 and improves the in vitro production and quality of bovine blastocysts

Effects of adding different concentrations of melatonin (10^?7, 10^?9 and 10^?11 M) to maturation (Experiment 1; Control, IVM  + 10^?7, IVM  + 10^?9, IVM  + 10^?11) and culture media (Experiment 2; Control, IVC  + 10^?7, IVC  + 10^?9, IVC  + 10^?11) were evaluated on in vitro bovine embryonic development. The optimal concentration of melatonin (10^?9 M) from Experiments 1–2 was tested in both maturation and/or culture media of Experiment 3 (Control, IVM  + 10^?9, IVC  + 10^?9, IVM /IVC  + 10^?9). In Experiment 1, maturated oocytes from Control and IVM  + 10^?9 treatments showed increased glutathione content, mitochondrial membrane potential and percentage of Grade I blastocysts (40.6% and 43%, respectively). In Experiment 2, an increase in the percentage of Grade I blastocysts was detected in IVC  + 10^?7 (43.5%; 56.7%) and IVC  + 10^?9 (47.4%; 57.4%). Moreover, a lower number and percentage of apoptotic cells in blastocysts were observed in the IVC  + 10^?9 group compared to Control (3.8 ± 0.6; 3.6% versus 6.1 ± 0.6; 5.3%). In Experiment 3, the IVC  + 10^?9 treatment increased percentage of Grade I blastocysts with a lower number of apoptotic cells compared to IVM /IVC  + 10^?9 group (52.6%; 3.0 ± 0.5 versus 46.0%; 5.4 ± 1.0). The IVC  + 10^?9 treatment also had a higher mRNA expression of antioxidant gene (SOD 2) compared to the Control, as well as the heat shock protein (HSPB 1) compared to the IVM  + 10^?9. Reactive oxygen species production was greater in the IVM /IVC  + 10^?9 treatment group. In conclusion, the 10^?9 M concentration of melatonin and the in vitro production phase in which it is used directly affected embryonic development and quality.     

Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

The use of ultrasonography in the reproductive evaluation of boars

The objective was to study the use of ultrasound as a complementary test in the breeding soundness evaluation in male pigs and study the pattern of echogenicity of the testicular parenchyma in boars of different racial groups. Twenty‐six adult boars from four different racial groups were used, 10 from the Piau breed (group 1), four from the commercial and finishing group (group 2), six Pietrain breed (group 3) and six from the Duroc breed (group 4). All animals were evaluated for breeding soundness evaluation and the ultrasound examination of the testicles. The groups of animals that were evaluated showed no difference in the main semen parameters that were evaluated, except for the sperm volume, concentration of the ejaculated sperm and the supravital staining; the lowest figures were for the animals from the Piau breed (group 1). In relation to the testicular biometrics, Duroc animals (group 4) had a greater scrotal width compared to the other groups. But when we assessed the intensity of pixels of the testicles, there was a difference between groups. The groups 2 (finishing animals), 3 (Pietrain) and 4 had no difference between themselves. Group 3 had greater pixel intensity in relation to group 1. Of the 26 animals studied, five showed an abnormality during ultrasound evaluation, like hydrocele, hyperechoic mass in the testicular parenchyma, cyst in the head of the epididymis and the presence of fluid in the head and tail of the epididymis. The various animal groups studied did not differ in the principal reproductive parameters evaluated, showing that despite the great variability of reproductive traits between breeds and within the same breed, the breeding soundness evaluation, the more complete it is, is essential for the selection of breeders and the ultrasonography of the reproductive system becomes an important addition in this examination.     

Equine foetal gender determination in mid‐ to late gestational mares: A practical inquiry

In recent years, the interest in equine foetal gender determination (FGD) during gestation increased remarkably. Ultrasonographic FGD can be performed in two different periods during gestation. The earliest examination can take place at a gestational age of 60–70 days, whereby the genital tubercle is used to differentiate between male and female foeti. The time window of the second approach is wider (120–210 days), and there are more characteristics to take into consideration. In this article, the feasibility and accuracy of ultrasonographic FGD in mid‐ to late gestation are evaluated. One hundred twenty‐one mares from different breeds with a pregnancy stage between 120 and 270 days were examined once, using B‐scale ultrasonography (Esaote MyLab™ClassC). None of the mares were sedated nor shaved, and the procedure was completed within 15 min. Diagnosis was firstly based on the gonads. The final judgement was made based on all visible foetal reproductive organs. In three cases with a pregnancy stage beyond 257 days, FGD was not possible. All of the examined mares in which a FGD could be performed gave birth to a healthy foal. In 98% of the examinations (116/118), the diagnosis was correctly made. In both cases of misdiagnosis, only one characteristic was seen during the procedure and wrongly interpreted. Beyond 210 days of pregnancy, the extremities can preclude a good visualization of the inguinal region. In conclusion, equine FGD in mid‐ to late gestation is an accessible and accurate technique, although a good ultrasound device is a prerequisite and experience and expertise is necessary.     

Idiopathic eosinophilic colitis lesions of the equine small (descending) colon

Five horses with a primary surgical lesion of the small (descending) colon were diagnosed with eosinophilic colitis based on visual and histopathological examination. These were evident as visibly striking, hyperaemic, focal lesions of the small colon, with serosal petechiation, oedema and marked thickening of the intestinal wall at the site. Areas of focal necrosis were also evident. The gross appearance of the lesions were considered to be sufficiently severe in all cases to merit resection, due to concerns about intestinal necrosis and septic peritonitis. An inability to fully exteriorise the affected portion of intestine to perform a resection and anastomosis necessitated intraoperative euthanasia of one horse. A total of three horses survived to hospital discharge. Eosinophilic colitis lesions are a rare cause of severe small colon disease, but should be considered in cases with similar visual characteristics.     

Pedotransfer functions to assess adsorbed phosphate using iron oxide content and magnetic susceptibility in an Oxisol

Adsorbed phosphate in soils can be chemically extracted; however, this process is both time‐consuming and not cost‐effective if large numbers of samples have to be analysed. Indirect assessment of adsorbed phosphate by pedotransfer functions (PTFs) can help optimize fertilizer strategies. This study aimed to evaluate the spatial variability of adsorbed phosphate (P_ads), iron oxides and magnetic susceptibility (MS) in oxisols and to calibrate PTFs to predict P_ads. A total of 308 soil samples were collected from Hapludox and Eutrudox soils formed from sandstone in Brazil. The contents of clay (196–607 g/kg), iron oxides (40–165 g/kg), MS (1.2–29 × 10^?6 m³/kg) and P_ads (327–842 mg/kg) were in the range of typical values for these highly weathered soils. This study showed that the attributes studied were spatially dependent. Geomorphic surfaces enabled understanding of spatial variability and helped to develop a more efficient sampling scheme to calibrate PTFs. Moreover, the adsorbed phosphate in these oxisols could be predicted by a PTF using iron oxides and MS as predictors. The MS attribute enabled the most accurate prediction (concordance coefficient = 0.95, root‐mean‐square error = 46 mg/kg and relative improvement in root‐mean‐square error = ?4.12) of spatial variability through PTF compared to other predictors.     

Food resources and cyprinid diet in permanent and temporary Mediterranean rivers with natural and regulated flow

This study addresses the differences in food availability, diet and feeding activity of the Iberian barbel, between permanent and temporary nonregulated rivers, and the effect of flow regulation on feeding parameters. A total of 267 adult barbels were seasonally collected in four nonregulated and regulated rivers from permanent and temporary basins, and their gut content was analysed. Locally available food sources were evaluated across sites and seasons. Barbels from the permanent nonregulated river exhibit a more variable and diversified diet in which invertebrates assumed a large importance, especially during high flows. Barbels from the temporary nonregulated river presented a more uniform diet composed of plant material and detritus, particularly in drought seasons. Flow regulation affected different flow components in both systems, but the effects on food resources and barbels' diet were similar, resulting in an intra‐annual stabilisation of resource availability and fish diet, with a higher consumption of plants and detritus. Changes in fish diet and feeding activity in both nonregulated and regulated rivers were strongly associated with the seasonal variability of streamflow components, particularly between low‐ and high‐flow periods, and with the reduction in flow variability in the case of dam regulation. Results from this study can be used to improve guidelines for flow requirement implementation.     

Evaluation of the Effects of Different Stunning Methods on the Stress Responses and Meat Quality of the Amazon hybrid surubim,Pseudoplatystoma fasciatum female × Leiarius marmoratus male

Different stunning methods (carbon dioxide [CO₂] and hypothermia) used in industrial fish processing were compared with asphyxia in air, which is used in traditional fishing, to evaluate the effects of these methods on the stress responses and the meat quality of the Amazon hybrid surubim, Pseudoplatystoma fasciatum female × Leiarius marmoratus male. After the application, blood and behavioral indicators were evaluated. Following death by gill cutting, the fish were gutted, and analyses of muscle pH, rigor mortis, the exudation of the meat, and coloration were performed. Significantly higher levels of cortisol and glucose were exhibited by the fish subjected to asphyxia than by the fish stunned by either CO₂ or hypothermia. The fish subjected to asphyxia displayed decreasing muscle pH during the first 2 h after death and showed higher rates of rigor mortis after 3 h than the fish that were stunned by the other two methods. The asphyxia is a practice that exposes fish to suffering by causing an increase in stress responses, which affects meat quality. The immersion of fish in water and ice (hypothermia) for 5 min was more effective stunning method than the use of CO₂, resulting in higher loss of sensibility and greater welfare.     

Early resynchronization protocols for goats: Progestogens can be used prior to an early pregnancy diagnosis without affecting corpus luteum functionality

This study aimed to evaluate the exogenous progesterone (P4) effect on the luteal function from Day 16 to Day 21 of the oestrous cycle in inseminated goats with unknown pregnancy status. A total of 54 does passed through a short progestin-based synchronization protocol and, on Day 16 of the following oestrous cycle, 27 does received a new P4 device which was retained until Day 21. Blood samples were collected daily from all does during this period, as well as on Day 24. Pregnancy diagnoses were performed on Day 30. Serum P4 values from 26 animals (G_NPSP: Group of non-pregnant does with second sponge: n = 8; G_NPNSP: Group of non-pregnant does without second sponge: n = 6; G_PSP: Group of pregnant does with second sponge: n = 5; G_PNSP: Group of pregnant does without second sponge: n = 7) were determined by radioimmunoassay commercial kits. No P4 differences were found between groups (G_NPSP: 3.1 ± 2.8; 1.7 ± 1.8; 0.4 ± 1.0; and 0.0 ± 0.0 vs. G_NPNSP: 4.4 ± 1.8; 3.0 ± 2.2; 0.8 ± 0.8; and 0.0 ± 0.0 or G_PSP: 4.2 ± 1.0; 3.4 ± 0.6; 3.3 ± 1.6; 3.2 ± 0.9; 3.6 ± 1.2; 3.5 ± 1.3; 2.7 ± 1.3 vs. G_PNSP: 4.4 ± 1.6; 3.6 ± 1.5; 3.7 ± 1.5; 3.8 ± 1.4; 3.2 ± 1.2; 3.1 ± 1.2; 3.6 ± 1.1; D16, D17, D18, D19, D20, D21, D24, respectively) or for the interaction of group and time. In conclusion, a second progestogen device had no effect on luteolysis or early pregnancy in the following oestrous cycle.