Ensiling Characteristics of Sainfoin(Onobrychis viciifolia) addition with or without of Polyethylene glycol (PEG)
Description
Protein degradation usually happened during legume ensiling attributed to high (20%) protein contents. Condensed tannins have several bioactivity such as inhibit bacteria and fungi , decreased protein degradation by combine with protein , etc.Sainfoin (Onobrychis viciifolia) is a legume rich in condensed tannins.Several studies use polyethylene glycol (PEG) deactivated CT at ensiling to observed tannins impact.Thus objected to present study is to find out condensed tannins impact on Ensiling Characteristics of Sainfoin(Onobrychis viciifolia) addition with or without of Polyethylene glycol (PEG).Established 2 treatments (with and without addition of PEG) × 5 sampling time points (3,7,14,30,60 days of ensiling) × 3 replicates samples silages. Using two-way ANOVA for a 2×5(PEG and control treatments× five ensiling time days) to data analysis (IBM SPSS 22.0 software, New York, USA).The data collected from different fermentation days include contents of dry matter, dry matter loss, crude protein,Non-Protein Nitrogen, soluble protein, Neutral detergent -insoluble protein, Acid detergent -insoluble protein, Neutral detergent fiber, Acid detergent fiber, Extractable condensed tannins , Protein-bound condensed tannin, Fiber-bound condensed tannin, total condensed tannin, water soluble carbohydrates, ammonia nitrogen, lactic acid, acetic acid, propionic acid, Lactic acid bacteria, Aerobic bacteria, yeast , moulds.
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Silage samples were freeze dried and ground to 1.0mm-screen to obtain DM content. The WSC were determined by using boiling water to extracted according to anthrone method(Ren et al., 2020,https://doi.org/10.1016/j.biortech.2020.123238). Total nitrogen (TN) was determined on automatic Kjeldahl nitrogen analyzer (K9840, Hanon Co. Ltd, Shandong, china) according to the procedure described by AOAC (AOAC. 1999. Official methods of analysis of the association of official agricultural chemists, 16th ed, 5th revision. AOAC International, Gaithersburg, MD), then the CP can calculate as TN × 6.25.NDF and ADF contents were determined by using followed methods(Van Soest et al., 1991). The NPN, NDIP, ADIP were analyzed by followed methods(Licitra et al., 1996,https://doi.org/10.1016/0377-8401(95)00837-3),SOLP determined by Coomassie Brilliant Blue G-250 binding with protein (Bradford, 1976, https://doi.org/10.1016/0003-2697(76)90527-3). For analysis of fermentation characteristics in sainfoin. 20g wet sample from each fermented silage was taken, then combined with 180mL distill water and blended in a homogenizer (L-1BA, Kuansons Biotechnology Co. Ltd, shanghai, PR china). Using four layers of cheesecloth to filtered the mixture, the supernatant was collected for analyzed VFA and ammonia N content. The pH measured by using a portable pH meter (PHS-3C，Instrument and electrical science instrument Co. Ltd, shanghai, PR china). The ammonia nitrogen contents were determined by phenol-hypochlorite colorimetric method(Weatherburn, 1967, Phenol-Hypochlorite Reaction for Determination of Ammonia. Analytical Chemistry 39, 971–974.). For VFA contents, the supernatant was filtration through 0.22-μm dialyzer before analyzed on a High Performance Liquid Chromatography (HPLC) (1200 series, Agilent Technologies, Inc., Waldbronn, Germany). The analytical column was C-18(150×4.6mm, FMF-5559-EONU, FLM Scientific Instrument Co. Ltd, Guangzhou, PR china). The analyzed procedure as followed, mobile phase was Na₂HPO₄(1mM) with the flow rate 0.6 mL·min−1, oven temperature 50◦ C and injection volume was 20μL(Xu et al., 2020, https://doi.org/10.1111/1751-7915.13623).Use the methods described by Terrill et al., (1992,https://doi.org/10.1002/jsfa.2740580306)to analysis the concentrations of extractable, protein-bound, and fiber-bound CT in sainfoin, with the CT purified from whole-sainfoin plants as a standard. Microbial count was carried out according to the methods(He et al., 2020,https://doi.org/10.1016/j.biortech.2020.123496). Briefly, samples were homogenized in 90 mL sterilized saline then the supernatant was serially diluted. The Lactic acid bacteria was detected on Man Rogosa Sharpe (MRS) agar after incubated at 37◦C for 48–72 h. The aerobic bacteria was detected on nutrient agar after incubated at 30◦C for 24h. The yeasts and molds were detected on Rose Bengal agar after incubated at 30 ◦C for 78-120 h.