ABSTRACT
Objective: To improve the fertilizing ability of frozen buffalo semen using the more beneficial, accurate, and cheap technique of laser irradiation at specific wavelengths and exposure times. Materials and Methods: The red laser source (625 nm) was used in this study with 5 watts output power and for the irradiation of the semen samples for 5 min; the laser focus spot area was 1 cm2.Thirty straws belonging to five buffalo bulls were used in this study. Results: The results show that total motility (%) and progressive motility (%) increased insignificantly after 5 min of exposure (73.8 ± 1.4 and 60.4 ± 1.1, respectively) compared to the control sample (70.9 ± 0.9 and 57.5 ± 1.7, respectively). All velocity parameters (velocity average path,velocity curved line, and velocity straight line µm/sec) recorded a significant (p < 0.05) increase in samples measured 5 min after exposure (52.3 ± 1.3, 83.5 ± 2.0, and 43.5 ± 1.2, respectively) compared to the untreated ones (47.1 ± 2.0, 76.3 ± 3.1, and 38.6 ± 1.9, respectively). Conclusion: The application of the red laser light on buffalo semen postthawing resulted in a positive correlation with almost every motility parameter; it may be recommended to apply this technique pre-in vitro fertilization for embryo production of buffalo species.
ABSTRACT
This study was conducted to monitor the cellular and molecular changes of buffalo cumulus-oocytes complexes (COCs) cultured under high or low oxygen levels. Morphologically good quality COCs (n = 1627) were screened using brilliant cresyl blue (BCB) staining and placed into three groups (BCB+, BCB- and control). All groups of COCs were cultured under low (5%) or high (20%) oxygen tensions. Intracellular and molecular changes including oocyte ultrastructure, lipid contents, mitochondrial activity and transcript abundance of genes regulating different pathways were analyzed in the matured oocyte groups. The results revealed that oxygen tension did not affect cumulus expansion rates, however the BCB+ group had a higher (P ≤ 0.05) expansion rate compared with the BCB- group. BCB- oocytes recorded the lowest meiotic progression rate (P ≤ 0.05) under high oxygen levels that was linked with an increased level of reactive oxygen species (ROS) compared with the BCB+ oocytes. Ultrastructure examination indicated that BCB+ oocytes had a higher rate of cortical granules migration compared with BCB- under low oxygen tension. In parallel, our results indicated the upregulation of NFE2L2 in groups of oocytes cultured under high oxygen tension that was coupled with reduced mitochondrial activity. In contrast, the expression levels of MAPK14 and CPT2 genes were increased (P ≤ 0.05) in groups of oocytes cultured under low compared with high oxygen tension that was subsequently associated with increased mitochondrial activity. In conclusion, data from the present investigation indicated that low oxygen tension is a favourable condition for maintaining the mitochondrial activity required for nuclear maturation of buffalo oocytes. However, low-quality oocytes (BCB-) responded negatively to high oxygen tension by reducing the expression of gene-regulating metabolic activity (CPT2). This action was an attempt by BCB- oocytes to reduce the increased levels of endogenously produced ROS that was coupled with decreased expression of the gene controlling meiotic progression (MAPK14) in addition to nuclear maturation rate.
