Preparation of acacia tannin loaded lipid microparticles by solid-in-oil-in-water and melt dispersion methods, their characterization and evaluation of their effect on ruminal gas production In Vitro

Tannin extracts have wide biological activity in ruminant nutrition. The possibility of masking their bitter taste and enhancing sustained release in the rumen can be achieved through encapsulation. The objectives of this study were to prepare an encapsulated Acacia tannin extract (ATE) suitable for ruminants using the solid-in-oil-in-water (S/O/W) method, and to evaluate the microparticles in terms of morphology, encapsulation efficiency and in vitro release under varying pH. Subsequently, the effect of the microparticles on rumen in vitro gas and methane production would be evaluated. Lipid microparticles were prepared using the double emulsion process with palm oil and lard, dichloromethane, and Tween80/Span80 emulsifiers. The microparticles produced by S/O/W emulsion tended to be smaller (P = 0.06), and had greater encapsulation efficiency compared with those produced by the melt dispersion method. Scanning electron micrographs showed microparticles had stable cylindrical and spherical shapes, with mean size of 34± 10.2 μm. Maximum encapsulation efficiencies of 78.6% and 80.1% were obtained with lard and palm oil as lipid wall materials, respectively, even under high core material loading percentage of 80%. Wall material type did not affect the characteristics of microparticles. In acetate buffer, only about 20% of tannin was released from the lipid-encapsulated microparticles into buffer media after 24 hours. In contrast, about 90% of the tannin had been released into solution before eight hours in the crude extract. Lipid-encapsulated ATE reduced rumen gas and methane production in vitro (P ˂0.05) in both Eragrostis and total mixed ration (TMR) diet substrates, but the magnitude of reduction was lower than that obtained when unencapsulated ATE was the additive (10% vs 20% for total gas and 17% vs 24% for methane). Crude ATE and palm oil encapsulated ATE reduced the concentration of methane in sampled gas (P = 0.054) when fermenting the TMR substrate, but this effect was not observed in the Eragrostis substrate. Ammonia nitrogen concentration was greater in encapsulated ATE compared with the crude ATE (P ˂0.001). These results show that the lipid-encapsulated ATE produced small-sized and more uniform microparticles, with high encapsulation efficiency compared with microparticles prepared by melt dispersion. Encapsulation of ATE enhanced the sustained release of tannin in the rumen, and with the potential to improve gas production and reduce methane production.