Amaro Felipe X, Kim Donghyeon, Agarussi Mariele C N, Silva Vanessa P, Fernandes Tatiane, Arriola Kathy G, Jiang Yun, Cervantes Andres P, Adesogan Adegbola T, Ferraretto Luiz F, Yu Shukun, Li Wenting, Vyas Diwakar
Department of Animal Sciences, University of Florida, Gainesville, FL.
Universidade Federal de Vicosa, Vicosa, MG, Brazil.
Transl Anim Sci. 2020 Nov 28;5(1):txaa222. doi: 10.1093/tas/txaa222. eCollection 2021 Jan.
Two separate experiments were carried out to evaluate the effects of incremental doses of 10 exogenous endo-acting α-amylase and exo-acting glucoamylase; 1LAT (bacterial α-amylase), 2AK, 3AC, 4Cs4, 5Trga, 6Afuga, 7Fvga, and 10Tg (fungal α-amylases, glucoamylases, and α-glucosidase), 8Star and 9Syn (fungal amylase-mixtures; experiment 1) and three exogenous proteases; 11P14L, 12P7L, and 13P30L (bacterial proteases; experiment 2) on in vitro dry matter digestibility (IVDMD) and in vitro starch digestibility (IVSD) of mature dent corn grain using a batch culture system. Incremental doses of the exogenous enzymes (0, 0.25, 0.50, 0.75, and 1.00 mg/g of dried substrate) were applied directly to the substrate (0.5 g of ground corn, 4 mm) in sextuplicate (experiment 1) or quadruplicate (experiment 2) within F57 filter bags, which were incubated at 39 °C in buffered rumen fluid for 7 h. Rumen fluid was collected 2-3 h after the morning feeding from three lactating dairy cows and pooled. Cows were consuming a midlactation total mixed ration (TMR; 1.60 Mcal/kg DM and 15.4%; net energy of lactation and crude protein, respectively). Three independent runs were carried out for each experiment. Data were analyzed as a randomized complete block design using run as the blocking factor. Dose was used as a fixed factor while run was considered a random factor. Linear, quadratic, and cubic orthogonal contrasts were also tested. In experiment 1, enzymes 2AK, 3AC, and 10Tg did not increase ( > 0.10) IVDMD and IVSD, whereas 0.25 mg of enzymes 1LAT, 5Trga, and 8Star increased ( < 0.01) IVDMD by 23%, 47%, and 62% and IVSD by 35%, 41%, and 58%, respectively, compared with the control. Enzymes 4Cs4, 6Afuga, 7Fvga, and 9Syn linearly increased IVDMD and IVSD ( < 0.01). Greatest increases in IVDMD (82.9%) and IVSD (85.9%) resulted with 1 mg of 6Afuga compared to control. In experiment 2, the lowest dose of exogenous proteases 11P14L and 12P7L increased ( < 0.01) IVDMD by 98% and 87% and IVSD by 57% and 64%, respectively, whereas the highest dose of 13P30L increased ( = 0.02) IVDMD by 44.8% and IVSD by 30%, relative to the control. In conclusion, IVSD and IVDMD were increased by one α-amylase, certain glucoamylases, and all proteases tested, with the glucoamylase 6Afuga in experiment 1 and the neutral protease 12P7L in experiment 2, increasing IVDMD and IVSD to the greater extents. Future in vivo studies are required to validate these findings before these enzyme additives can be recommended for improving the digestibility of mature dent corn grain.
进行了两项独立实验,以评估递增剂量的10种外源性内切α-淀粉酶和外切葡糖淀粉酶;1LAT(细菌α-淀粉酶)、2AK、3AC、4Cs4、5Trga、6Afuga、7Fvga和10Tg(真菌α-淀粉酶、葡糖淀粉酶和α-葡萄糖苷酶)、8Star和9Syn(真菌淀粉酶混合物;实验1)以及三种外源性蛋白酶;11P14L、12P7L和13P30L(细菌蛋白酶;实验2)对成熟马齿玉米籽粒体外干物质消化率(IVDMD)和体外淀粉消化率(IVSD)的影响,采用批次培养系统。将递增剂量的外源性酶(0、0.25、0.50、0.75和1.00 mg/g干底物)直接应用于底物(0.5 g磨碎的玉米,4 mm),在F57滤袋中一式六份(实验1)或一式四份(实验2),于39℃在缓冲瘤胃液中孵育7小时。在早晨喂食后2 - 3小时从三头泌乳奶牛收集瘤胃液并混合。奶牛采食的是泌乳中期全混合日粮(TMR;分别为1.60 Mcal/kg DM和15.4%;泌乳净能和粗蛋白)。每个实验进行三次独立运行。数据采用随机完全区组设计进行分析,以运行作为区组因素。剂量作为固定因素,而运行被视为随机因素。还测试了线性、二次和三次正交对比。在实验1中,酶2AK、3AC和10Tg未提高(>0.10)IVDMD和IVSD,而0.25 mg的酶1LAT、5Trga和8Star使IVDMD分别提高(<0.01)23%、47%和62%,IVSD分别提高35%、41%和58%,与对照相比。酶4Cs4、6Afuga、7Fvga和9Syn使IVDMD和IVSD呈线性增加(<0.01)。与对照相比,1 mg的6Afuga使IVDMD(82.9%)和IVSD(85.9%)增加最多。在实验2中,外源性蛋白酶11P14L和1十二P7L的最低剂量使IVDMD分别提高(<0.01)98%和87%,IVSD分别提高57%和64%,而最高剂量的13P30L使IVDMD提高(=0.02)44.8%,IVSD提高30%,相对于对照。总之,一种α-淀粉酶、某些葡糖淀粉酶和所有测试的蛋白酶均提高了IVSD和IVDMD,实验1中的葡糖淀粉酶6Afuga和实验2中的中性蛋白酶12P7L使IVDMD和IVSD提高的程度更大。在这些酶添加剂被推荐用于提高成熟马齿玉米籽粒的消化率之前,需要进行进一步的体内研究来验证这些发现。