Fibrin–alginate hydrogel supports steroidogenesis,in vitro maturation of oocytes and parthenotes production from caprine preantral follicles cultured in group

This study aimed to establish a culture system that improves the in vitro development of caprine preantral follicles. In a first experiment, follicles were encapsulated as a single unit per bead and cultured singly or in groups or with five follicles in the same alginate (ALG) bead for 18 days. In a subsequent experiment, the “five follicles per bead” design was chosen to culture in ALG, fibrin–alginate (FA) or hyaluronate (HA) for 18 days. In a third experiment, we chose the five follicles per bead in FA to culture for 30 days. The culture set‐up of five follicles per ALG bead increased antrum formation and follicle diameter compared to the other culture designs (p < .05). Moreover, under this condition, 44.44% of the oocytes from in vitro cultured preantral follicles reached meiotic resumption. A significant increase of follicle diameter occurred in attachment system and FA (p < .05), but the ALG condition reached the highest among all groups on day 18 (p < .05). Follicles encapsulated in matrix produced more estradiol and progesterone than attachment system (p < .05). The expression of MMP‐9 mRNA was higher in FA than in other groups (p < .05) and similar to antral follicles from in vivo control (p > .05). Only FA group resulted in oocytes matured. After 30 days, oocytes from preantral follicles in vitro grown in FA developed to eight‐cell parthenotes. In conclusion, a culture system using FA supported the development of caprine preantral follicles cultured in group and included in the same bead of hydrogel, improving the oocyte maturation and producing parthenotes.     

Effectiveness of α-tocopherol and selenium supplements in preventing lupinosis-associated myopathy in sheep

Objective To compare the effectiveness of combined selenium and α-tocopherol acetate treatments in preventing lupinosis-associated myopathy in sheep.
Design Measurement of plasma muscle and liver enzymes, and histopathological examination of muscle and liver in the treatment groups.
Procedure The treatments were: subcutaneous injections of selenomethionine and vitamin E (sc(SeM+E)) , an intraruminal selenium pellet and oral doses of vitamin E and intramuscular injections of selenomethionine combined with either oral doses of vitamin E (imSeM+orE) or intramuscular injections of vitamin E in an oily carrier. Another group received no supplements, while a control group was given selenium pellets on day 0 and fortnightly oral doses of vitamin E from day 0 to 72. To produce lupinosis-associated myopathy, the sheep were fed a diet low in vitamin E and given repeated injections of a crude extract of Diaphorthe toxica. Groups sc(SeM+E) and imSeM+orE were stressed by dosing with protected polyunsaturated fatty acids from day 56 onwards.
Results Lupinosis-associated myopathy was induced in all unsupplemented sheep. In these sheep the storage of Se increased and that of vitamin E decreased. The subcutaneous treatment was highly effective in preventing lupinosis-associated myopathy and also produced the highest vitamin E concentrations in plasma and liver. Supplemental vitamin E was more efficacious than supplemental Se. Concentrations of vitamin E in the livers of sheep given intramuscular vitamin E were higher than expected based on plasma concentrations. Oral doses of vitamin E proved the least effective method of increasing concentrations in liver. Lupinosis did not affect Se concentrations in liver or muscle.
Conclusion The sc(SeM+E) treatment is highly effective in preventing lupinosis-associated myopathy but needs to be further assessed when selenium and vitamin E are both limiting in the diet.