Resumption of Cyclic Ovarian Activity by Herbal Preparation AyuFertin in Bulgarian Murrah Buffaloes at Early Postpartum.

This study evaluates the phytochemical composition and ability of herbal preparation AyuFertin, supplemented shortly after calving, to resume the cyclic ovarian activity in postpartum Bulgarian Murrah buffaloes. In total, 13 healthy Bulgarian Murrah buffaloes at the age of 4-10 years, calved in March-April 2019 were involved in the experiment. Seven experimental buffaloes were treated individually per os with AyuFertin according to producer instruction. All animals were subjected to regular ultrasound examination of ovaries. GC-MS analysis of fatty acids composition and HPLC-DAD analysis of carotenoid and tocopherol content in AyuFertin were conducted. The determination of estrus occurrence and natural mating were done by a fertile bull. The development of the large follicles (12.5-14.3 mm) in 85.7% of treated animals was observed on day 24 postpartum compared to 0% in controls. Clinical signs of estrus were recorded in 71.4% of the animals, followed by an 80% pregnancy rate versus 16% in controls within 70 days after calving. In conclusion, the supplementation of the bioactive herbal product AyuFertin from day 21 postpartum can stimulate the follicular growth in the buffalo's ovary, which leads to the development of follicles with ovulatory capacity, followed by successful ovulation. The above-mentioned treatment resulted in a better pregnancy rate in the treated multiparous compared to the control buffaloes bred by natural service.

Positive effect of natural antioxidant oregonin from Alnus incana bark on ram semen quality stored at 5 °C for 48 h.

The maintenance of high vitality and motility of ram's spermatozoa during storage at low temperatures has a crucial role for successful fertilization. This study evaluates the effect of the natural antioxidant oregonin on ram semen quality, stored at 5 °C for 48 h. Еighteen ejaculates (three repetitions for 6 ejaculates) from three local breed rams, collecting by artificial vagina, with volume > 1 ml, concentration > 1 × 109/ml and mass motility >3.5 were used for chilling. Each ejaculate was separated in two equal parts, diluted with Tris-glucose-glycerol-egg yolk extender with no oregonin or supplemented with 100µÐœ oregonin until adjustment of the sperm concentration to 200 × 106 cells/ml and stored at 5 °C for 48 h. The semen quality assessment was based on the main kinematic (by CASA analysis), morphological parameters (by BrightVit kit staining) and mitochondrial status (by MitoView staining) of the spermatozoa on 0, 24 and 48 h of storage, and on in vivo fertility test. Oregonin did not impair the morphology and kept sustained motility of ram spermatozoa stored at 5 °C for 48 h. The curvilinear velocity indicated faster movement of the oregonin treated sperms that corresponded with high percent of spermatozoa with active mitochondria in these samples. The fertilizing capacity of spermatozoa was preserved and pregnancy rate in the experimental group was 80% versus 60% in control. In conclusion, this study provides a new data about positive effect of the natural antioxidant oregonin, supplemented to the extender, on chilled ram semen quality, including fertilizing ability.

Oregonin from Alnus incana bark affects DNA methyltransferases expression and mitochondrial DNA copies in mouse embryonic fibroblasts.

Oregonin is an open-chain diarylheptanoid isolated from Alnus incana bark that possesses remarkable antioxidant and anti-inflammatory properties, inhibits adipogenesis, and can be used in the prevention of obesity and related metabolic disorders. Here, we aimed to investigate the effects of oregonin on the epigenetic regulation in cells as well as its ability to modulate DNA methylating enzymes expression and mitochondrial DNA (mtDNA) copies. Our results show that oregonin altered the expression of DNA methyltransferases and mtDNA copy numbers in dependency on concentration and specificity of cells genotype. A close correlation between mtDNA copy numbers and mRNA expression of the mtDnmt1 and Dnmt3b was established. Moreover, molecular modeling suggested that oregonin fits the catalytic site of DNMT1 and partially overlaps with binding of the cofactor. These findings further extend the knowledge on oregonin, and elucidate for the first time its potential to affect the key players of the DNA methylation process, namely DNMTs transcripts and mtDNA.