Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia–pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest^® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.     

Sperm subpopulations influence the pregnancy rates in cattle

This study aimed to study the characteristics and subpopulations of spermatozoa from bulls with low and high reproductive performance based on pregnancy rates. Based on historical records of pregnancy rate from four farms, 24 bulls were selected. Two groups were established, with low pregnancy rates (n = 12; LOW), including bulls that presented pregnancy rates <52.27% (33.33% to 51.81%); and a group with high pregnancy rates (n = 12; HIGH), with pregnancy rates >52.27% (52.27% to 69.64%), after fixed-time artificial insemination (FTAI). The thawed sperm straws were analysed to sperm kinetics, morphology, plasma membrane integrity and sperm subpopulations. The LOW group exhibited a higher proportion of static cells (p < .05). In contrast, the HIGH group showed greater percentages for membrane integrity and total and progressive motility, and cells with fast and medium velocity (p < .05). In the cluster procedures, four sperm subpopulations were established. The low-fertility bulls presented the highest percentage of subpopulation 2 (41.46%), characterized by slow and progressive spermatozoa. The high-fertility bulls exhibited the highest percentage of subpopulation 3 (37.17%), characterized by fast and nonlinear spermatozoa. Results from this study indicated that bulls with greater percentages of fast and nonlinear spermatozoa seem to have greater fertilization capacity and the subpopulations analysis can be considered a tool to identify ejaculates with high fertility.     

Use of dimethylformamide to cryopreserve alpaca semen previously incubated with collagenase

The objective of this study was to evaluate the effect of collagenase and two final dimethylformamide (DMF) concentrations (4% and 7%) on alpaca frozen-thawed sperm quality. A total of 25 ejaculates from 5 alpaca were obtained using electroejaculation. Each individual ejaculate was evaluated and then diluted 4:1 in a solution of 1 mg/ml collagenase in HEPES-TALP medium and incubated for 4 min at 37°C. Subsequently, samples were diluted in TRIS-fructose-citric acid-egg yolk and cooled to 5°C. Then, each sample was divided in two aliquots and DMF at final concentration of 4% or 7% was added, equilibrated for 1 hr at 5°C and frozen over liquid nitrogen vapours. A Kruskal–Wallis test was used to evaluate the sperm morphometry, and Completely Random Block designs were used to analyse sperm motility, viability, membrane function and acrosome status. After collagenase incubation, none of the samples showed thread formation, and sperm parameters were preserved. Non-progressive motile sperm were higher (p < .05) in equilibrated samples (4% DMF: 31.8 ± 8.3% and 7% DMF: 36.3 ± 11.8%) compared to raw (10.1 ± 4.3%) and frozen-thawed semen (4% DMF: 9.7 ± 1.8% and 7% DMF: 7.5 ± 3.2%). Sperm membrane function, membrane integrity and intact acrosomes were higher (p < .05) in raw semen (40.1 ± 12.2%, 94.6 ± 3.2% and 91.3 ± 8.1%) compared to frozen-thawed samples (4% DMF: 19.8 ± 4.7%, 53.2 ± 2.7%, 65.7 ± 8.7% and 7% DMF: 20.4 ± 4.5%, 54.1 ± 1.4%, 64.6 ± 9.1%). Length of the sperm head was lower in frozen-thawed samples, being statistically different with 4% DMF compared to pre-freezing samples. The ratio between acrosome and head areas was greater (p < .05) in frozen-thawed samples. Incubation of raw alpaca semen with collagenase decreased the thread formation without affecting sperm quality. Frozen of collagenase treated alpaca semen with 4% or 7% DMF did not preserve the sperm parameters in thawed samples.     

Free-range system and supplementation of 25-hydroxicholecalciferol increases the performance and serum vitamin levels in mixed-parity sows

Improvements in sow productivity have raised questions regarding dietary vitamin D recommendations. The present study aimed to evaluate the effects of the housing system with access to sunlight exposure and supplementation of 25-hydroxicholecalciferol on performance and serum levels of 25(OH)D₃ in sows during gestation and lactation. Sows were distributed in an experimental design with two housing systems: gestation crates or gestation free-range system with external area for sunlight exposure; and two diets: 0 or 50 μg of 25-hydroxicholecalciferol kg⁻¹. The use of 25-hydroxicholecalciferol tended (P = 0.052) to improve total born and influenced (P = 0.046) on number of born alive. Litter weight at birth was also increased (P = 0.01) by 25-hydroxicholecalciferol supplementation; 25-hydroxicholecalciferol supplementation and housing system (free-range with sunlight exposure) tended to increase weaning weight (P = 0.07) and litter daily gain (P = 0.051) during lactation. Exposure to sunlight and 25-hydroxicholecalciferol supplementation increased 25(OH)D₃ serum levels when compared with control treatment during gestation (136.95 vs. 113.92 ng mL⁻¹; P = 0.035) and lactation (120.29 vs. 88.93 ng mL⁻¹; P = 0.026). In conclusion, the association of 25-hydroxicholecalciferol supplementation with exposure to sunlight during gestation improved significantly 25(OH)D₃ serum levels and consequently performance traits in gestation and lactation.     

Cryopreservation of ram semen: Manual versus programmable freezing and different lengths of equilibration

The aim of our study was to examine effects of the length of semen equilibration as well as two freezing techniques on ram sperm post-thaw quality. The ejaculates of Wallachian sheep rams (n = 12) were collected by an electro-ejaculation, equilibrated in a Triladyl® (0, 2, 4, 6, and 8 h) containing glycerol and egg yolk and frozen by programmable freezing (PF) or manual freezing (MF). After thawing, sperm samples were subjected to the motility (computer-assisted sperm analysis [CASA]), viability (SYBR-14/PI), and fertilizing ability (FA) (in vitro penetration/fertilization test on bovine oocytes) assays. It was found that the equilibration of 6 h (E-6) ensured higher post-thaw sperm motility and progressive movement compared with other lengths tested, irrespective of a freezing technique. The E-6 sperm viability did not differ between PF and MF but was lower (P < 0.05) than control. Sperm FA (E-6) was similar in PF (60.44%) and MF (62%) but slightly lower than in fresh (72.8%). Our data demonstrate that the use of MF was comparable with PF, which can be applied in the field conditions without need in a piece of cost-expensive equipment, which can greatly benefit the gene bank of animal genetic resources.     

Effect of the Nixtamalization Process on the Dietary Fiber Content,Starch Digestibility,and Antioxidant Capacity of Blue Maize Tortilla

Nixtamalization is an ancient process developed by the Mesoamerican cultures. Initially, volcanic ashes were used and then calcium hydroxide in commercial production, and more recently nixtamalization with calcium salts (NCS) has been proposed. The aim of this study was to evaluate the effect of NCS on carbohydrate digestibility and antioxidant capacity in the elaboration of blue maize tortillas. NCS in blue tortillas showed a high amount of total dietary fiber (14.27 g/100 g), the main fraction being insoluble dietary fiber. The contents of resistant starch and slowly digestible starch did not change with the nixtamalization process. The predicted glycemic index value was lower in blue tortillas with the NCS process (58) than with the traditional nixtamalization process (71). In general, NCS in blue tortillas presented a higher antioxidant capacity than traditional tortillas (ferric reducing antioxidant power method), indicating that phenolics present in blue maize maintain their activity after cooking. It can be concluded that the nutraceutical features (high dietary fiber content and antioxidant capacity) of blue maize tortillas are enhanced when they are elaborated with the NCS process.     

Addition of Sodium Stearoyl Lactylate to Corn and Sorghum Starch Extrudates Enhances the Performance of Pregelatinized Beer Adjuncts

Double mashing for wort production is a time‐consuming process that can be reduced if pregelatinized adjuncts are used. Optimal extruding conditions were determined to obtain brewing adjuncts from corn and sorghum starch. For corn starch extrusion, a Box–Behnken design was devised in which moisture, screw speed, temperature of the barrel, and concentrations of sodium stearoyl lactylate (SSL) were varied, and sorghum starch was extruded according to a 2³ model in which the modified variables were moisture, SSL concentration, and temperature. The aim was to maximize starch damage and minimize resistant starch and final viscosity as determined with a Rapid Visco Analyzer. The treatments that satisfied these requirements were mashed, and wort extract yield was determined. Glucose, maltose, and maltotriose concentrations in the resulting worts were determined by HPLC with a refractive index detector. Feedstock tempering and SSL content were the most important factors affecting the response; for corn starch, treatments with lower moisture (20%) and middle levels of SSL (0.5%) or with high levels of both moisture (40%) and SSL (1%) produced the most desirable samples for mashing, whereas for sorghum starch the best treatment was tempering to 20% moisture and containing middle levels of SSL (0.5%). No statistical differences were found between these experimental treatments and the control.     

A SURVEY OF RADIOLOGISTS AND REFERRING VETERINARIANS REGARDING IMAGING REPORTS

An imaging report is a vital communication tool between a radiologist and clinician. In a field where in‐person communication may not be readily available, it is imperative that the report clearly relays pertinent clinical information in a timely manner. The purpose of this observational study was to describe and compare opinions and expectations of small animal general practitioners, veterinary specialists, and veterinary radiologists regarding the imaging report. Online surveys were distributed, and data were collected from 202 veterinary clinicians and 123 veterinary radiologists. The majority (89%) of clinicians were satisfied with their imaging reports and stated that they read the radiology report as soon as it was available (92%). Just less than half (48%) of clinicians indicated it was standard of care that a board‐certified veterinary radiologist read all imaging studies. Radiologists and clinicians agreed that a clinical history (98% and 94%, respectively) and clinical question (82% and 68%, respectively) were needed to generate a good radiology report. Fifty‐five percent to 70% of clinicians prefer red bulleted reports, which included incidental findings (96%); while radiologists slightly favored prose reporting (37–46%). Clinicians found it helpful when additional imaging (86%), medical (71%), and surgical recommendations (73%) were made. About one‐third of specialists who had been in practice for >11 years thought they were better able to interpret imaging for their own specialty than the radiologist. Clinicians voiced discontentment with reports that were not completed in a timely manner or did not give a prioritized differential list. Further studies are warranted to provide a more in‐depth evaluation of veterinary radiology reporting structure and style.