ABSTRACT
ABSTRACT: Piper betle L. is edible plant richer in polyphenols that might improve feed utilization in rumen diet. The objective of the present study was to investigate the effect of various Piper betle L. powder (PL) doses on in vitro rumen microorganisms, ruminal biogas and fermentation end-product production, and biohydrogenation including lipolysis-isomerization. The completely randomized design used five levels of PL supplementation (0, 25, 50, 75 and 100 mg DM) incubated with 400 mg of a basal substrate of Pangola hay and concentrate (50:50). The matrix compounds (g/kg DM) of 0.27 catechin, 0.11 rutin, 3.48 quercetin, 0.41 apigenin, 0.04 myricetin, 0.27 kaempferol, 0.76 eugenol and 0.22 caryophyllene derived from PL altered the fermentation pattern, with an increase in degradable nutrients and total volatile fatty acids and acetogenesis without shifting pH during fermentation. These values promoted in vitro gas production, with higher carbon dioxide and lower methane production. Although, hydrogen recovery from lipolysis-isomerization in biohydrogenation was limited, PL successfully promoted stearic acid (C18:0) accumulation by changing the biohydrogenation pathway of fatty acids, causing more C18:1 trans-11 rather than C18:2 trans-11, cis-15. Consequently, this resulted in more conjugated linoleic acid (CLA) cis-9, trans-11, CLA trans-10, cis-12 and CLA trans-11, cis-13. Enhanced PL supply increased total bacteria and fungal zoospores due to a reduction in rumen protozoa. In conclusion, our results demonstrated that PL is a feed additive with potential for ruminants, promising improved ruminal fermentation and biohydrogenation, while reducing methane production.
RESUMO: Piper betle L. é uma planta comestível rica em polifenois que podem melhorar a utilização de alimentos na dieta de ruminantes. O objetivo do presente estudo foi investigar o efeito de várias doses de Piper betle L em . pó (PL) sobre microrganismos do rúmen in vitro, biogás ruminal e produção de produtos finais de fermentação e bio-hidrogenação, incluindo lipólise e isomerização. O delineamento inteiramente casualizado utilizou cinco níveis de suplementação de PL (0, 25, 50, 75 e 100 mg de MS) incubados com 400 mg de um substrato basal do feno de Pangola e concentrado (50:50). Os compostos da matriz (g / kg MS) de 0,27 catequina, 0,11 rutina, 3,48 quercetina, 0,41 apigenina, 0,04 miricetina, 0,27 kaempferol, 0,76 eugenol e 0,22 cariofileno derivado de PL, alteraram o padrão de fermentação com o aumento de nutrientes degradáveis e voláteis totais, ácidos graxos e acetogênese sem alterar o pH durante a fermentação. Esses valores promoveram a produção de gás in vitro, com maior dióxido de carbono e menor produção de metano. Embora a recuperação de hidrogênio da lipólise-isomerização na bio-hidrogenação tenha sido limitada, o PL promoveu com sucesso o acúmulo de ácido esteárico (C18: 0) alterando a via de bio-hidrogenação dos ácidos graxos, causando mais C18: 1 trans-11 do que C18: 2 trans-11, cis -15. Consequentemente, isso resultou em mais ácido linoléico conjugado (CLA) cis-9, trans-11, CLA trans-10, cis-12 e CLA trans-11, cis-13. O suprimento aprimorado de PL aumentou o total de bactérias e zoósporos de fungos devido a uma redução no número de protozoários do rúmen. Em conclusão, nossos resultados demonstram que o PL é um aditivo alimentar com potencial para ruminantes, prometendo fermentação ruminal e bio-hidrogenação aprimoradas, enquanto reduz a produção de metano.
ABSTRACT
A reversed-phase high-performance liquid chromatography with diode-array detection (HPLC-DAD) was developedand validated to estimate the phenolic acids (gallic acid, caffeic acid, syringic acid, p-coumaric acid, sinapic acid, andferrulic acid), flavonoids (catechin rutin, myricetin, quercetin, apigenin, and kaempferol), ascorbic acid, and eugenol.The chromatogram condition was set in suitable wavelength 272 nm and run flow rate 0.7 µl/minutes using HPLCAgilent Technologies 1260 Infinity, a reversed-phase Zorbax SB-C18 column (3.5 µm particle size, i.d. 4.6 mm × 250mm) with the mobile phase solution (1:9, HPLC-grade acetonitrile:1% acetic acid). The linearity, precision, limit ofdetection, limit of detection, and accuracy were R2 > 0.9907, relative standard deviation < 1%, 0.005 µg/ml, 0.015 µg/ml, and 96%–102%, respectively. As a result, all the selective compounds were successfully separated, identified, andquantified. The enormous contents were found in quercetin and eugenol, expressing crude content (mean, 5.989 mg/g)and residue content (mean, 1.934 mg/g) for quercetin, while crude content (mean, 3.209 mg/g) and residue content(mean, 0.184 mg/g) for eugenol. Consequently, this method could be applied, repeated, and developed for the laterobservation, especially in commercially inclination of Piper betle analysis