Our experiment demonstrated an extension of Percoll ® Density Gradient Centrifugation to use as a selection tool for sperm sexing developed for dairy farmer. We develop a simple protocol for bovine sperm sexing and used for artificial insemination (AI) and in vitro fertilization (IVF) for embryo transfer (ET). The quality of sperm after centrifugation in each layer were significantly different (P <0.01). There was no significant difference between percentage of live sperm after centrifugation and fresh sperm before centrifugation (P >0.05). The phenotype and swimming capacity of sperms in each layer were significantly different (P < 0.01). It was found that the most appropriate sperm in the layer after centrifugation and after quinacrine staining was between 65-70% Percoll ® . X-bearing spermatozoa at this layer was 60.75% and motility was 95.86±0.46%. After insemination, the pregnancy ratio in the experiment was 40% which is similar to the percentage of normal AI. The number of female offspring in the experiment was 71.4% in selected sperm vs 50% in non-selected sperm AI. In the In Vitro fertilization experiment, sperm from our 7 layer Percoll ® gradient concentration can fertilize and produce blastocyst embryo successfully. The simple 7 layer Percoll ® gradient concentration protocol is appropriate for dairy farmer to do as a pre-AI step to increase their female offspring in their herd.

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Peroxidation damage to spermatozoa and seminal plasma has an important role in sperm quality. Thus, the objective of this study was to determine the levels of lipid and protein oxidation in spermatozoa and seminal plasma of Asian elephants (Elephas maximus) with varying percentage of progressive motility. Lipid and protein oxidation was measured by the thiobarbituric acid-reactive species (TBARS) assay and the 2, 4-dinitrophenylhydrazine (DNPH) carbonyl groups assay, respectively. Fresh semen samples were collected from Asian elephants and classified according to the percentage of motile spermatozoa into good (>60%) and poor (≤20%) motility. Results revealed that seminal plasma malondialdehyde (MDA) and seminal plasma protein carbonyls (PCs) were significantly higher in poor motility than in good motility (p < .05). The MDA and PC levels in seminal plasma were negatively correlated with the percentages of progressive motility (p < .05). In addition, the negative correlation between sperm concentration and seminal plasma MDA level was investigated (p < .05). The sperm viability was also negatively correlated with sperm PC level (p < .05). This study indicated that lipid and protein oxidation has deleterious effect on semen quality of Asian elephants.

The objective of this study was to determine the effect of bovine follicular fluid proteins (bFF) and their differently charged groups as maturation media supplements for in vitro embryo development. bFF was obtained by aspiration from large healthy follicles (4–10 mm in diameter) and was precipitated by 30–50% (NH4)2SO4. The precipitated protein was fractionated into basic and acidic fractions by ion-exchanger columns. In experiment 1, the oocytes were matured in TCM-199 with 1) FBS+hormones (control) and 2) 10% bFF. The oocyte maturation rate, the development to the blastocyst stage rate and blastocyst cell number were not significantly different between the groups. However, the INFα and IGF-2r expression levels in the 10% bFF were higher than in the control (P<0.05). In experiment 2, the specific charge proteins of bFF (basic and acidic) were also used as media supplements in the maturation medium. The basic fraction had higher oocyte maturation rate and blastocyst cell number when compared with addition of acidic fraction (P<0.05). The expression levels for almost all developmentally important genes in the basic fraction were greater than those in the acidic fraction, particularly INFα (P<0.05). Most of the protein in the basic fraction was associated with the immune response and mRNA processing. In conclusion, supplementation of 10% bFF alone in maturation medium can support oocyte maturation and embryo development. The basic fraction in bFF seemed to have effect on oocyte maturation rate and blastocyst cell number.

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Nowadays, there have been attempts to use phytochemicals in fruits to reduce the risk of suffering a given sickness. In this work, we studied the potential effects of mango (cultivar “Nam Dok Mai”) and banana (cultivar “Khai”) to attenuate DNA oxidative damage in MCF‐10A cells induced by 4‐hydroxyestradiol (4‐OHE2). The effects of mango extract (MNE) and banana extract (BKE) were comparable with three carotenoid compounds, β‐carotene, lycopene, and lutein. The oxidative‐induced DNA damage was evaluated by 8‐hydroxy‐2‐deoxyguanosine (8‐OHdG) reduction. 4‐OHE2‐induced DNA oxidative damage in MCF‐10A cells showed a decrease in 8‐OHdG formation when treated with MNE and BKE. Both fruit extracts also enabled the regaining production of Phase II detoxifying (GSTs and NQO1) and antioxidant (SOD, GPx, and CAT) enzymes during 4‐OHE2‐induced DNA oxidative damage in the MCF‐10A cells when compared with the untreated control. These results indicate that MNE and BKE can exert potential mitigating effects against 4‐OHE2‐induced DNA oxidative damage in MCF‐10A cells by enhancing the activities of detoxifying and antioxidant enzyme. Practical applications Long‐term exposure to estrogen increases the risk of sickness since oxidative stress via the estrogen pathway, leading to DNA damage. This study indicated that mango (cultivar “Nam Dok Mai”) extract contains β‐carotene and lycopene, while banana (cultivar “Khai”) extract contains β‐carotene and lutein, which act as natural antioxidants. Both fruit extracts have preventive properties against oxidative DNA damage and are potentially good supplements for women taking E2 between HRT.