Comparison of five different methods to assess the concentration of boar semen

Both for research and practical purposes, accurate and repeatable methods are required to assess the concentration of boar semen samples. Since the method which is used may influence the results considerably, the aim of the present study was to compare 5 frequently used techniques to determine boar semen concentration. Fifty ejaculates were collected from 37 different boars at an artificial insemination centre. Subsequently, each ejaculate was analyzed for sperm concentration by means of 2 different types of colorimeters (Colorimeter 1: Model 252, Sherwood Scientific Ltd, Cambridge, UK; Colorimeter 2: Ciba-Corning, Schippers, Bladel, The Netherlands), the Burker counting chamber (golden standard), and the Hamilton Thorne Analyzer (Ceros 42.1) using 2 types of Leja chambers (the 'former' and the 'recently developed'). Each ejaculate was assessed 5 times with each of the 5 methods, and the repeatability, expressed by coefficient of variation (CV), was determined for each method. The different methods were compared using Pearson's correlations and limits of agreement. The colorimeters yielded the lowest CV's (both 3.7%), while the former Leja chamber resulted in the highest CV (12.4%). Moreover, significant (P0.71) were found between the results obtained by the different methods. The limits of agreement plots showed that none of the methods consistently over- or underestimated the sperm concentrations when compared to the Burker chamber, although there was a tendency toward higher over- or underestimation in highly concentrated sperm samples. Based on our results, there were no major differences in the assessment of sperm concentration between the evaluated methods. The choice of method used in a laboratory could therefore be based on factors such as cost, number of samples to be assessed and practical use, without thereby negatively affecting the validity of the results thus obtained

Detailed motility evaluation of boar semen and its predictive value for reproductive performance in sows

Reliable estimates of boar fertility potential from semen evaluation could be a valuable tool for boar selection. The aim of this study was to investigate the morphology and the detailed motility parameters of diluted boar semen and to relate these to their predictive value concerning conception and farrowing rate, litter size and the number of live born piglets. In addition, the optimal time for evaluation of the motility of preserved semen with respect to its predictive effect on fertility was determined. One hundred ejaculates from 38 boars were evaluated morphologically by eosin-nigrosin staining and different motility characteristics were assessed using Computer Assisted Semen Analysis (CASA). The motility was determined at 15, 45 and 120 minutes after incubation at 37 degrees C. The conception rate, farrowing rate, litter size and number of live born piglets were registered from 276 sows inseminated with these ejaculates. Different regression models were used to evaluate the predictive value of the semen characteristics on these fertility parameters, taking into account the effect of herd, parity and weaning to estrus interval. The motility characteristics of the spermatozoa varied significantly during the 15 to 120 minutes of incubation. The longer the incubation time, the more the velocity parameters along the actual cell path decreased, while the parameters of straightforward movement increased. The predictive value of individual semen parameters on conception and farrowing rate was very small. The predictive value of certain associations of different semen parameters, on the other hand, was significant. The percentage of motile spermatozoa had a significant (P<0.05) and positive effect on the total number of piglets born (litter size) and on the number of live born piglets, independent of the time of measurement (X-2 0.38-1.00 and 0.41-1.00, respectively). Accurate evaluation of the motility of a semen dose is therefore imperative for estimating its predictive value relating to fertility. In conclusion, since the time of evaluation after warming the samples significantly influences the motility parameters, CASA measurement should be done when the cells are completely acclimatized to 37 degrees C. On the basis of the available data, a 45 min incubation period appeared to be sufficient. The percentage of motile spermatozoa, as assessed by CASA on diluted semen, offers detailed predictive information regarding litter size, irrespective of the time of measurement

In vitro production of bovine embryos derived from individual donors in the Corral® dish

Background: Since the identity of the embryo is of outmost importance during commercial in vitro embryo production, bovine oocytes and embryos have to be cultured strictly per donor. Due to the rather low yield of oocytes collected after ovum pick-up (OPU) per individual cow, oocyte maturation and embryo culture take place in small groups, which is often associated with inferior embryo development. The objective of this study was to improve embryonic development in small donor groups by using the Corral (R) dish. This commercial dish is designed for human embryo production. It contains two central wells that are divided into quadrants by a semi-permeable wall. In human embryo culture, one embryo is placed per quadrant, allowing individual follow-up while embryos are exposed to a common medium. In our study, small groups of oocytes and subsequently embryos of different bovine donors were placed in the Corral (R) dish, each donor group in a separate quadrant. Results: In two experiments, the Corral (R) dish was evaluated during in vitro maturation (IVM) and/or in vitro culture (IVC) by grouping oocytes and embryos of individual bovine donors per quadrant. At day 7, a significantly higher blastocyst rate was noted in the Corral (R) dish used during IVM and IVC than when only used during IVM (12.9% +/- 2.10 versus 22.8% +/- 2.67) (P < 0.05). However, no significant differences in blastocyst yield were observed anymore between treatment groups at day 8 post insemination. Conclusions: In the present study, the Corral (R) dish was used for in vitro embryo production (IVP) in cattle; allowing to allocate oocytes and/or embryos per donor. As fresh embryo transfers on day 7 have higher pregnancy outcomes, the Corral (R) dish offers an added value for commercial OPU/IVP, since a higher blastocyst development at day 7 is obtained when the Corral (R) dish is used during IVM and IVC

Micromechanical Analysis of the Hyaluronan-Rich Matrix Surrounding the Oocyte Reveals a Uniquely Soft and Elastic Composition

The cumulus cell-oocyte complex (COC) matrix is an extended coat that forms around the oocyte a few hours before ovulation and plays vital roles in oocyte biology. Here, we analyzed the micromechanical response of mouse COC matrix by colloidal-probe atomic force microscopy. We found that the COC matrix is elastic insofar as it does not flow and its original shape is restored after force release. At the same time, the COC matrix is extremely soft. Specifically, the most compliant parts of in vivo and in vitro expanded COC matrices yielded Young's modulus values of 0.5 ± 0.1 Pa and 1.6 ± 0.3 Pa, respectively, suggesting both high porosity and a large mesh size (≥100 nm). In addition, the elastic modulus increased progressively with indentation. Furthermore, using optical microscopy to correlate these mechanical properties with ultrastructure, we discovered that the COC is surrounded by a thick matrix shell that is essentially devoid of cumulus cells and is enhanced upon COC expansion in vivo. We propose that the pronounced nonlinear elastic behavior of the COC matrix is a consequence of structural heterogeneity and serves important functions in biological processes such as oocyte transport in the oviduct and sperm penetration

Direct comparison of distinct naive pluripotent states in human embryonic stem cells

Stem cells & developmental biolog