Accurate semen evaluation is necessary to maintain high reproductive efficiency but difficult to accomplish. The objective was to determine if the ability to bind oviduct cells or oviduct glycans are useful supplements to traditional semen analyses. Measuring binding to specific soluble glycans is less laborious than assessing binding to oviduct cell aggregates and more suitable for routine use. Previous work has shown that sperm binding to oviduct cells improves fertility prediction, possibly by estimating the ability of sperm to form an oviduct reservoir. The two oviduct glycan motifs, biantennary 6-sialylated N-acetyllactosamine (bi-SiaLN) and Lewis X trisaccharide (Le X), that bind boar spermatozoa with high affinity and specificity were tested. Semen from 30 boars was shipped overnight for laboratory analysis and for inseminations to determine fertility (n = 3 replicates). Oviduct cell binding and traditional sperm analyses including motility and morphology were completed. Additionally, binding to soluble fluoresceinated glycans bi-SiaLN, sulfated Le X (suLe X), and the control lactosamine disaccharide (LacNAc) was measured. Inseminations were at 15 farms (>50 matings per boar) in the Midwest and farrowing data from all matings were used. Pregnancy rate (PR) and litter size (LS) were adjusted to account for different farms, number of services, number of doses inseminated, and sow parity, using the MIXED procedure in SAS 9.4. A fertility index (FI) was generated, consisting of PR × LS, to estimate boar overall fertility. Finally, the GLMSELECT procedure was used to select variables having a significant impact on PR, LS, and FI. The predictive models constructed were further analyzed using the REG procedure and accounted for 58% or more of the variation in PR, LS, and FI [PR (P < 0.001, r 2 = 0.60), LS (P < 0.001, r 2 = 0.58), and FI (P < 0.001, r 2 = 0.63)]. The final model for PR includes oviduct cell binding as well as boar age, % distal droplets, head morphology, tail morphology, beat/cross frequency, and curvilinear velocity. The final model for LS includes boar age, % distal droplets, tail morphology, and overall morphology. Finally, the FI model included boar age, % distal droplets, head morphology, tail morphology, curvilinear velocity, and semen volume per ejaculate. Although binding to intact oviduct cells was impactful as a means to predict PR, binding to specific soluble oviduct glycans was not a useful supplement to traditional semen analysis.

No abstract

Compared to control the diluted/dynamised FSH addition increased progesterone production but decreased the estradiol production after in vitro culture of isolated porcine preantral follicles. Taken together the results suggest that at least for progesterone production the mechanism of action of diluted/dynamised FSH differs from its vehicle.

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Recent research has focused on developing secondary, preantral follicles as a new way to collect female gametes to develop in vitro or to study folliculogenesis. Secondary follicles are abundant in ovaries of almost all females, and can be isolated from the ovarian cortex of fresh or cryopreserved tissue. These follicles must be matured to obtain mature oocytes, but the mechanism for early follicle development is not well understood. Currently, culture techniques are being developed to promote the growth and quality of these follicles. The aim of this preliminary study was to compare 2 embedding systems for porcine follicles to improve preantral follicle culture. We collected secondary preantral follicles isolated from porcine ovaries recovered from the abattoir. Ovaries were transported to our laboratory in saline solution (0.9% NaCl) and cut into smaller fragments with a scalpel. Follicles were further mechanically isolated with a 23 1/2-gauge needle and placed into a dish with medium TCM199 plus Hepes (Lonza 12–117F) supplemented with 5% fetal bovine serum (FBS). Secondary follicles less than 300 µm were selected. Furthermore, only clear follicles with dark oocytes and no antrum were used. The selected follicles were randomly divided into 5 different treatment groups: 1) embedded in 0.5% alginate gel; 2) 1.0% alginate gel; 3) 1.0% agarose gel; 4) 2.0% agarose gel; and 5) control. Three follicles were placed in each well with 280 µL of α-minimal essential medium supplemented with 3.5 μg mL–1 of insulin 10 μg mL–1 of transferrin, 100 μg mL–1 of l-ascorbic acid, and 7.5% porcine serum and cultured for 4 days at 39°C at 5% CO2. Media (140 µL) from each well was changed on Day 2 and saved for metabolic and functional analysis. Initial diameters and measurements on Day 2 and 4 were taken to analyse dimensional growth. We evaluated 6 follicles per group per replicate, for a total of 3 replicates (18 per group). All recorded parameters were subjected to a 2-way ANOVA using the procedure of the Generalized Linear Model (SPSS, 18th version, SPSS Inc., Chicago, IL, USA). Independent variable was the result of the balance between the size on Day 2 and 0 and the size on Day 4 and 0 time of observation. Data were normally distributed. Least squares means tests were used to perform statistical multiple comparisons. The α level was set at 0.05. All data were expressed as quadratic means and mean standard errors. The results in the Table 1 show that at Day 2 there was no difference between groups. However, after 4 days of development, the group agar 2% and alginate 1% showed a statistical difference when compared with the control. These results suggest that there is a positive effect when the follicles are embedded during culture. Table 1.Growth of Follicles (in μm) since Day 0 in different embedding systems