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.

Objective: This study evaluated the phytochemical compounds, antioxidant, and antimicrobial activities of Vernonia amygdalina (VA) leaf extract and used it as an auxiliary herbal constituent in natural hair shampoo. Methods: VA leaf was macerated in various solvents. All crude extracts were phytochemical compounds screened and determined the antioxidant and antimicrobial activities before formulating a natural shampoo formula. Natural shampoos were evaluated with physic-chemical properties and sensory satisfaction. Results: About 95% ethanolic extract was the most suitable substrate for product development. It reveals the inhibition zone from 7.00±0.00 to 15.00±1.00 mm and possesses a broad spectrum of antibacterial agents against both Gram-positive and Gram-negative bacteria. The MICs and minimum bactericidal concentrations value ranged from 6.250 to 100 mg/ml and 25 to >200 mg/ml, respectively. About 95% ethanolic extract also revealed high antioxidant activity. The IC50 value of DPPH of 95% ethanolic extract was 1.88±0.02 μg/ml (% inhibition of 85.73±0.01) and the ferric reducing ability power was 23.00±0.50 mg AAE/100 gDW. These high biological activities may be due to a broad range of phytochemical compounds, including saponins, tannins, flavonoids, terpenoids, steroids, cardiac glycosides, and alkaloids. Conclusions: The VA leaf was suitable to use as an ingredient in natural shampoos with a low detrimental effect on normal skin flora. The shampoo with 0.00188% of 95% ethanolic extract was the most suitable formula in terms of characteristics and stability. This formula also obtained the highest satisfaction level under our sensory evaluation.

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Aims:The occurrence of bacterial disease in shrimp ponds is a major problem faced in shrimp farming. Thus, the aims of this study were to isolate and evaluate antibiotic resistant profile of Vibrio harveyi strain isolated from shrimp pond water, as well as to study the potential anti-Vibrio activity of Combretum quadrangulare Kurz. (CQ) and Mimosa pudica (MP) leaves extracts. Methodology and results: Vibrio harveyi WSC103 was isolated from water in white shrimp (Litopenaeus vannamei) culture pond and identified using 16S rRNA gene sequencing analysis. This strain showed characteristics of multidrugresistant (7 antibiotics). It had become more sensitive to antibiotics (9 out of 10 antibiotics) after plasmid curing. It is showed CQ and MP leaves extracts contain potent bioactive compounds (tannins, flavonoids, steroids, cardiac glycosides and alkaloids) against V. harveyi WSC103. The aqueous, 95% ethanolic and 75% acetone extracts of CQ (MIC value of 3.13-12.50 mg/mL) and MP (MIC value of 3.13-25.00 mg/mL) leaves revealed strong vibriostatic activity, but aqueous and 95% ethanolic extracts in both plants showed vibriocidal activity. The 95% ethanolic extract of both CQ and MP leaves displayed the excellent vibriocidal property with MBC value of 100 mg/mL with zone of inhibition at 11.44 ± 1.01 and 11.78 ± 1.01 mm by agar disc diffusion. Conclusion, significance and impact of study: The isolated Vibrio harveyi WSC103 was successfully characterized as a novel multidrug-resistant strain. The ethanolic C. quadrangulare Kurz. and M. pudica extracts exhibited prominent vibriostatic and vibriocidal capacities. These finding is proven that C. quadrangulare Kurz. and M. pudica extracts would be an alternative anti-Vibrio agent for aquaculture infectious treatment.