Abstract
The present study was conducted to investigate the effect of omega-3 poly unsaturated fatty acid (PUFA), α-linolenic acid (ALA; 18:3 n-3) on the in vitro maturation (IVM) of buffalo oocytes and subsequent embryonic development. Buffalo cumulusoocyte complexes (COCs; n = 2282) were in vitro matured in TCM-199 (0.6% fatty acid free bovine serum albumin, 0.02 Units/ml FSH, 1 µg/ml 17-β-estradiol, 10 µg/ml epidermal growth factor, 50 µg/ml gentamicin) supplemented with 0 (control), 25, 50, 100, 150 or 300 µm ALA under an atmosphere of 5% CO2 in air at 38.5oC for 22-24 h. The matured oocytes were then fertilized in Tyrode’s Albumin Lactate Pyruvate (TALP) medium and cultured in synthetic oviductal fluid (SOF) medium. Concentrations up to 100 μm ALA improves (P ≤ 0.05) the cumulus expansion compared to control. Higher percentage of oocytes reaching MII stage was observed at 50 μm and 100 μm of ALA compared to control (P ≤ 0.05). Concentrations of 150 and 300 µm ALA were detrimental both for cumulus expansion and nuclear maturation rate of buffalo oocytes. Moreover, supplementation with 100 μm ALA improved (P ≤ 0.05) cleavage rate compared to control and treatment with 50 and 100 μm ALA yielded significantly higher morulae compared to control. The results of present study indicate that the supplementation with 100 μm ALA to the IVM medium improves nuclear maturation rate of buffalo oocytes and subsequent early embryonic development.
Highlights
The Nili-Ravi buffalo is considered to be one of the best milk producers among other breeds of buffaloes in the world (Warriach et al, 2008)
Supplementation with 50 and 100 μM ALA resulted in increased rate of cumulus expansion (P ≤ 0.05) compared to control
The percentage of oocytes remaining at germinal vesicle breakdown (GVBD) and MI stage were recorded higher in 300 μM ALA (26.2% ± 2.2 and 39.1% ± 1.4) compared to control (17.7% ± 3.1 and 20.7% ± 2.4), respectively
Summary
The Nili-Ravi buffalo is considered to be one of the best milk producers among other breeds of buffaloes in the world (Warriach et al, 2008). This breed has the potential of producing more than 5000 liters of milk /lactation (Bilal et al, 2006), the average milk yield is quite low, which raises the opportunities of genetic improvement through assisted reproductive technologies like artificial insemination, and multiple ovulation and embryo transfer (MOET). Several studies have evaluated different types of media (Totey et al, 1993; Abdoon et al, 2001; Zicarelli et al, 2003), and its supplements including different sources of protein (Chauhan et al, 1998), hormones (Totey et al, 1993), thiol compounds (Gasparrini et al, 2006; Zicarelli et al, 2005), growth factors (Purohit et al, 2005) and antioxidants (Ullah et al, 2006) for in vitro maturation of buffalo oocytes with variable success rates and blastocyst production rate of no more than 10-20% (Kumar and Anand, 2012) which is relatively lower when compared to 30-40 % blastocyst production rate in cattle (Rizos et al, 2008)
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