Abstract
Moringa oleifera seed extract was added to the in vitro maturation media of bovine oocytes as an alternative antibiotic to evaluate its effect on the oocyte maturation and its antimicrobial activity. Oocytes (n = 405) were recovered from ovaries from slaughtered cows, classified by quality and divided in four categories according to the cytoplasm appearance and cumulus cells layers. Oocytes were washed four times in TCM-199 supplemented with fetal bovine serum (FBS) and FSH, then placed in maturation media, which contained TCM-199 supplemented with FBS, FSH and the corresponding treatments: TCM-199+gentamicin (Control), TCM-199 without antibiotic (Test), moringa (M) 1.0, 5.0 and 10.0 mg/mL. After 22 h at 38.5°C in a 5% CO2 humidified atmosphere, oocytes were examined to determine cumulus expansion (as categorical results yes or no) and cumulus expansion Index (CEI). Additionally, 400 µL of used maturation media was retrieved and stored for the antimicrobial activity tests in agar cultures. Evaluation of the cultures for colony-forming units (CFU) was done at 24 and 48 h post culture. Results were analyzed with Chi Square test (for expansion rates) and an ANOVA (for the CEI and CFU) using the SAS system, data are presented as mean ± standard error. Statistically, there was no difference between treatments (P >0.05). Numerically M 1.0 and Test obtained the highest maturation rate (96.43%; 96.20%). M 1.0 and Control showed the highest CEI (2.26±0.21; 2.25±0.26). For the CFU count, Control showed the lowest result at 24 and 48 h post culture (0.00 ± 0.00; 0.11 ± 0.33), as M 10.0 showed the highest result at both 24 and 48 h (2.00 ± 3.60; 2.55 ± 4.58). The addition M 1.0 mg/mL to the maturation media is an alternative for the use of conventional antibiotics without decreasing maturation rates.
70 Effects of the addition of Moringa oleifera seed extract to in vitro maturation of bovine oocyte media and its antimicrobial activity
