韓奇鵬，張佩華，羅 玲，揭紅東，周傳社，陳 亮，孔志偉

(1.中國(guó)科學(xué)院亞熱帶農(nóng)業(yè)生態(tài)研究所亞熱帶農(nóng)業(yè)生態(tài)過程重點(diǎn)實(shí)驗(yàn)室，湖南省畜禽健康養(yǎng)殖工程技術(shù)中心，農(nóng)業(yè)部中南動(dòng)物營(yíng)養(yǎng)與飼料科學(xué)觀測(cè)實(shí)驗(yàn)站，湖南 長(zhǎng)沙 410125；2.湖南農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)技術(shù)學(xué)院畜禽遺傳改良湖南省重點(diǎn)實(shí)驗(yàn)室，湖南 長(zhǎng)沙 410128)

動(dòng)物生產(chǎn)層

不同來源吸附載體大豆糖蜜對(duì)奶牛瘤胃內(nèi)環(huán)境的影響

韓奇鵬1，2，張佩華2，羅 玲2，揭紅東2，周傳社1，陳 亮1，孔志偉1

(1.中國(guó)科學(xué)院亞熱帶農(nóng)業(yè)生態(tài)研究所亞熱帶農(nóng)業(yè)生態(tài)過程重點(diǎn)實(shí)驗(yàn)室，湖南省畜禽健康養(yǎng)殖工程技術(shù)中心，農(nóng)業(yè)部中南動(dòng)物營(yíng)養(yǎng)與飼料科學(xué)觀測(cè)實(shí)驗(yàn)站，湖南 長(zhǎng)沙 410125；2.湖南農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)技術(shù)學(xué)院畜禽遺傳改良湖南省重點(diǎn)實(shí)驗(yàn)室，湖南 長(zhǎng)沙 410128)

為考察不同來源吸附載體大豆糖蜜對(duì)奶牛瘤胃內(nèi)環(huán)境的影響，采用3×3拉丁方試驗(yàn)設(shè)計(jì)，選用3頭年齡相同(3胎次)、體重相近[(500±50) kg]且安裝永久性瘤胃瘺管的中國(guó)荷斯坦奶牛為試驗(yàn)動(dòng)物，分別飼喂基礎(chǔ)日糧、豆皮糖蜜和麩皮糖蜜日糧。結(jié)果表明，豆皮糖蜜組和麩皮糖蜜組瘤胃pH均極顯著高于對(duì)照組(P<0.01)，而豆皮糖蜜組和麩皮糖蜜組之間瘤胃pH沒有顯著差異(P>0.05)；麩皮糖蜜組瘤胃NH3-N濃度顯著低于對(duì)照組(P<0.05)，但與豆皮糖蜜組沒有顯著差異(P>0.05)；各處理間奶牛瘤胃各單個(gè)揮發(fā)性脂肪酸(volatile fatty acid，VFA，包括乙酸、丙酸、異丁酸、丁酸、異戊酸和戊酸)濃度、總VFA濃度和乙丙比均沒有顯著差異(P>0.05)；但與對(duì)照組相比，豆皮糖蜜組瘤胃總VFA濃度有降低趨勢(shì)；而麩皮糖蜜組瘤胃總VFA濃度有提高趨勢(shì)。結(jié)果顯示，在奶牛飼料中可用豆皮糖蜜替代15%玉米以及用麩皮糖蜜替代6%玉米和10%麩皮，均具有調(diào)控瘤胃穩(wěn)衡的作用，且麩皮糖蜜替代效果優(yōu)于豆皮糖蜜。

大豆糖蜜；瘤胃內(nèi)環(huán)境；奶牛

大豆糖蜜是以低溫脫脂豆粕為原料，采用酸提和醇提等方法生產(chǎn)大豆?jié)饪s蛋白過程中，經(jīng)萃取、濃縮而成的純天然副產(chǎn)物[1-3]。大豆糖蜜中一般含有固形物50%、蛋白質(zhì)6%、灰分3%、脂肪5%和碳水化合物36%，成分較為復(fù)雜，是多種植物性化合物的集合體[4]，可用于提取大豆異黃酮、大豆低聚糖、皂甙、大豆胰蛋白酶抑制素等活性成分[5-8]；同時(shí)還可以利用微生物發(fā)酵生產(chǎn)丁醇、槐脂、乳酸等[8-11]。目前，對(duì)大豆糖蜜及其多活性成分在動(dòng)物營(yíng)養(yǎng)領(lǐng)域的研究處于起步階段。隨著大豆糖蜜添加量的增加，肉羊瘤胃菌體蛋白濃度不同程度升高；同時(shí)，大豆糖蜜可提高肉羊瘤胃液丙酸濃度和血液總蛋白濃度，降低丙酸轉(zhuǎn)氨酶濃度；提高肉羊日增重和飼料轉(zhuǎn)化率[12-15]。添加6%大豆糖蜜可促進(jìn)魯山牛腿山羊日增重和飼料轉(zhuǎn)化率[16]。梁麗莉和趙海明[17]在奶牛日糧中添加3 kg大豆糖蜜，泌乳高峰期奶牛體重、產(chǎn)奶量和乳脂率均有所增加，在玉米較缺乏的地區(qū)向奶牛日糧中添加大豆糖蜜可替代部分玉米。上述研究均是在動(dòng)物日糧中直接添加液態(tài)大豆糖蜜，而本研究利用具有吸附和固定作用的豆皮和麩皮作為大豆糖蜜的吸附載體，對(duì)大豆糖蜜進(jìn)行吸附后再添加至奶牛日糧中，考察其對(duì)奶牛瘤胃內(nèi)環(huán)境的影響，以期為實(shí)施反芻家畜瘤胃穩(wěn)衡狀態(tài)調(diào)控和促進(jìn)奶牛養(yǎng)殖產(chǎn)業(yè)健康可持續(xù)發(fā)展提供理論依據(jù)和技術(shù)支撐。

1 材料與方法

1.1 試驗(yàn)動(dòng)物及飼養(yǎng)管理

選擇3頭體況良好、年齡相同(3胎次)、體重相近[(500±50)kg]、安裝永久性瘤胃瘺管的中國(guó)荷斯坦奶牛為試驗(yàn)動(dòng)物。分別于06:00和15:00等量飼喂，自由飲水。

1.2 試驗(yàn)日糧

試驗(yàn)日糧參照NRC(2004)中國(guó)奶牛飼養(yǎng)標(biāo)準(zhǔn)配制[18]。試驗(yàn)日糧中分別添加的豆皮糖蜜和麩皮糖蜜每千克均能吸附300 g的大豆糖蜜(干物質(zhì)計(jì))，豆皮糖蜜和麩皮糖蜜均由豐益(上海)生物技術(shù)研發(fā)中心有限公司提供。各組日糧精飼料組成及營(yíng)養(yǎng)成分如表1所示。

1.3 試驗(yàn)設(shè)計(jì)

采用3×3拉丁方試驗(yàn)設(shè)計(jì)，進(jìn)行3期動(dòng)物試驗(yàn)。每期試驗(yàn)15 d，包括14 d預(yù)飼期和1 d樣品采集。

1.4 樣品采集與分析測(cè)定

分別于每期試驗(yàn)第15天的05:00、06:00、07:00、08:00、10:00、12:00、15:00、16:00、18:00、21:00和23:00采集瘤胃內(nèi)容物50 mL。用pH計(jì)(REX PHS-3C，上海儀器設(shè)備廠)測(cè)定其pH值后，4層紗布過濾[20]。再5 000 r·min-1離心10 min去除原蟲和飼料大顆粒，取上清液，采用比色法(UV2450紫外可見分光光度計(jì)，Shimadzu Corporation，Japan)測(cè)定氨氮(NH3-N)濃度[21-22]。另取1.5 mL上清液分裝于預(yù)先裝有25%偏磷酸的離心管內(nèi)搖勻，采用氣相色譜法測(cè)定乙酸、丙酸、丁酸等揮發(fā)性脂肪酸(VFA)含量[23]。色譜條件：色譜柱為Agilent TC-C18柱(250 nm×4.6 nm，i. d. 5 μm)，流速為0.6 mL·min-1，進(jìn)樣體積20 μL，紫外檢測(cè)波長(zhǎng)210 nm；柱溫為室溫。流動(dòng)相經(jīng)0.22 μm纖維膜過濾，超聲波脫氣[24]。

表1 試驗(yàn)?zāi)膛Ｈ占Z精飼料組成及其營(yíng)養(yǎng)成分Table 1 Ingredients and chemical composition of concentrate of the experimental diets

注：2)預(yù)混料成分組成如下：MgSO4·H2O 113.87 g·kg-1，MnSO4·H2O 9.54 g·kg-1，ZnSO4·H2O 9.61 g·kg-1，F(xiàn)eSO4·7H2O 2.69 g·kg-1，CuSO4·5H2O 2.55 g·kg-1，CoCl2·6H2O 181.68 mg·kg-1，Na2SeO3112.80 mg·kg-1，KI 58.86 mg·kg-1，VA 500 000 IU·kg-1，VD 60 000 IU·kg-1，VE 2 000 IU·kg-1。 3)產(chǎn)奶凈能為估測(cè)值[19]。

Note: 2)DDGS means distillers dried grains with soluble. 2)Premix components were as follows: MgSO4·H2O 113.87 g·kg-1, MnSO4·H2O 9.54 g·kg-1, ZnSO4·H2O 9.61 g·kg-1, FeSO4·7H2O 2.69 g·kg-1, CuSO4·5H2O 2.55 g·kg-1, CoCl2·6H2O 181.68 mg·kg-1, Na2SeO3112.80 mg·kg-1, KI 58.86 mg·kg-1, vitamin A (VA) 500 000 IU·kg-1, vitaminD (VD) 60 000 IU·kg-1, vitaminE (VE) 2 000 IU·kg-1. 3)NEL was estimated value[19].

1.5 試驗(yàn)數(shù)據(jù)統(tǒng)計(jì)及分析

試驗(yàn)數(shù)據(jù)采用SAS(8.2版)的MIXED過程統(tǒng)計(jì)，不同瘤胃發(fā)酵物的差異采用CONTRAST語句進(jìn)行比較。各時(shí)間點(diǎn)作為重復(fù)，統(tǒng)計(jì)差異顯著性定義為P<0.05，P>0.05為差異不顯著。

2 結(jié)果與分析

2.1 大豆糖蜜對(duì)奶牛瘤胃pH值和NH3-N濃度的影響

豆皮糖蜜組和麩皮糖蜜組瘤胃pH值均極顯著高于對(duì)照組(P<0.01)，而豆皮糖蜜組和麩皮糖蜜組之間瘤胃pH值沒有顯著差異(P>0.05)(表2)。麩皮糖蜜組瘤胃NH3-N濃度顯著低于對(duì)照組(P<0.05)，但與豆皮糖蜜組沒有顯著差異(P>0.05)，且豆皮糖蜜組與對(duì)照組之間瘤胃NH3-N濃度亦沒有顯著差異(P>0.05)。

表2 大豆糖蜜對(duì)奶牛瘤胃pH值和NH3-N濃度的影響Table 2 Effects of soybean molasses on pH value in the rumen and NH3-N concentration in dairy cows

注：同行數(shù)據(jù)不同小寫字母表示差異顯著(P<0.05)，不同大寫字母表示差異極顯著(P<0.01)。下表同。

Note: Different lower case or capital letters within the same row indicate a significant difference at the 0.05 or 0.01 levels, respectively.

2.2 大豆糖蜜對(duì)奶牛瘤胃VFA濃度的影響

各處理間奶牛瘤胃各單個(gè)VFA(乙酸、丙酸、異丁酸、丁酸、異戊酸和戊酸)濃度、總VFA濃度和乙丙比(A∶P)均沒有顯著差異(P>0.05)(表3)。但與對(duì)照組相比，豆皮糖蜜組瘤胃總VFA濃度有降低趨勢(shì)，較對(duì)照組降低了5.04%；而麩皮糖蜜組瘤胃總VFA濃度有提高趨勢(shì)，較對(duì)照組提高了10.34%。

表3 大豆糖蜜對(duì)奶牛瘤胃VFA濃度的影響Table 3 Effects of soybean molasses on ruminal VFA concentration in dairy cows

3 討論

3.1 大豆糖蜜對(duì)瘤胃pH的影響

瘤胃pH是評(píng)價(jià)瘤胃發(fā)酵狀況的綜合性指標(biāo)，它主要受日糧性質(zhì)，唾液分泌及瘤胃內(nèi)酸、堿性物質(zhì)排除、吸收的影響，決定著瘤胃微生物對(duì)底物的發(fā)酵利用效率。大豆(Glycinemax)糖蜜中低聚糖的主要發(fā)酵產(chǎn)物是乙酸、丙酸和丁酸，且這幾種是反芻動(dòng)物最重要的揮發(fā)性脂肪酸，約占瘤胃發(fā)酵總揮發(fā)性脂肪酸的95%以上[25]。本研究顯示，各組瘤胃pH值在6.52-6.68內(nèi)變化，均處在正常生理范圍內(nèi)[14]，且試驗(yàn)組間差異不顯著，表明試驗(yàn)?zāi)膛２墒澈蠖固敲鄣娜占Z后，瘤胃自身具有的緩沖能力仍能維持瘤胃內(nèi)pH值處于正常范圍之內(nèi)。

3.2 大豆糖蜜對(duì)瘤胃NH3-N濃度的影響

NH3-N是由瘤胃代謝中外源蛋白質(zhì)和內(nèi)源含氮物質(zhì)降解產(chǎn)物氨基酸脫氫基而產(chǎn)生，是瘤胃中主要微生物體系生長(zhǎng)的氮源[26-27]，其在瘤胃中的濃度也是體現(xiàn)微生物對(duì)氮源降解和利用的一個(gè)動(dòng)態(tài)指標(biāo)，受飼料蛋白的溶解度、瘤胃壁吸收和食糜排空速度的影響。本研究顯示，與對(duì)照組相比，豆皮糖蜜組奶牛瘤胃內(nèi)NH3-N濃度呈降低趨勢(shì)，而麩皮糖蜜組奶牛瘤胃內(nèi)NH3-N濃度顯著降低(P<0.05)，這可能是由于大豆糖蜜中存在的低聚糖加速了飼料蛋白質(zhì)的分解，致使NH3-N濃度增加；但同時(shí)又由于促進(jìn)了瘤胃微生物對(duì)所分解生成的NH3的利用率提高并合成菌體蛋白，從而導(dǎo)致瘤胃內(nèi)NH3-N濃度下降[14]；還可能是由于吸附后的豆皮糖蜜和麩皮糖蜜蛋白質(zhì)含量(13.25%和19.81%，實(shí)測(cè)值)及其蛋白組分尤其是瘤胃可降解蛋白和非降解蛋白比值之間的差異所致。

3.3 大豆糖蜜對(duì)瘤胃揮發(fā)性脂肪酸的影響

揮發(fā)性脂肪酸是指含有1～6個(gè)碳鏈的短鏈脂肪酸，其提供的能量占反芻動(dòng)物所需的70%～80%[28]，是反芻動(dòng)物生長(zhǎng)發(fā)育的直接能量來源之一。揮發(fā)性脂肪酸的主要成分有乙酸、丙酸、異丁酸、丁酸、異戊酸、戊酸等。大豆糖蜜中含有59%～62%的大豆低聚糖，具有較多的可消化物質(zhì)，而大豆低聚糖的主要發(fā)酵產(chǎn)物是乙酸、丙酸和丁酸。本研究表明，各試驗(yàn)組瘤胃揮發(fā)性脂肪酸濃度和乙丙比均沒有顯著差異，但與對(duì)照組相比，豆皮糖蜜組瘤胃總揮發(fā)性脂肪酸濃度有降低趨勢(shì)，而麩皮糖蜜組瘤胃總VFA濃度有提高趨勢(shì)。這可能是由于本研究應(yīng)用的豆皮糖蜜和麩皮糖蜜中大豆糖蜜的吸附濃度均為30%所致。

4 小結(jié)

在奶牛飼料中可用豆皮糖蜜替代15%玉米以及用麩皮糖蜜替代6%玉米和10%麩皮，均具有調(diào)控瘤胃穩(wěn)衡的作用，且麩皮糖蜜替代效果優(yōu)于豆皮糖蜜。根據(jù)大豆糖蜜主要功能物質(zhì)大豆異黃酮、大豆皂甙、低聚糖的作用，可以進(jìn)一步對(duì)奶牛生產(chǎn)性能和乳品質(zhì)等關(guān)鍵指標(biāo)進(jìn)行相應(yīng)的研究。

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(責(zé)任編輯 武艷培)

Effects of wheat bran and bean hull as adsorption substrates for soybean molasses on the ruminal environment of dairy cows

Han Qi-peng1,2, Zhang Pei-hua2, Luo Ling2, Jie Hong-dong2,Zhou Chuan-she1, Chen Liang1, Kong Zhi-wei1

(1.Key Laboratory for Agro-ecological Processes in Subtropical Region, Hunan Research Center of Livestock &Poultry Sciences, South-Central Experimental Station of Animal Nutrition and Feed Science of Ministry of Agriculture, Institute of Subtropical Agriculture, The Chinese Academy of Science, Changsha 410125, China;2. Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China)

We examined the effects of soybean molasses adsorbed to different substrates on the ruminal environment of dairy cows. Three Chinese Holstein dairy cows (all the same age, and three parity) with an average body weight of (500±50) kg were allocated three dietary treatments in a 3×3 Latin square design. The cows were fitted with the permanent ruminal fistula, the three experimental diets were similar except that some wheat bran (replaced 15% of corn meal by adsorbed soybean molasses with wheat bran) or soybean hull (replaced 10% of wheat bran and 5% corn meal by adsorbed soybean molasses with soybean hull). The results showed that the molasses that were adsorbed to soybean hull and wheat bran significantly increased the pH value of the rumen compared to the control group (P<0.01). However, there was no significant difference (P>0.05) between the pH values of the rumens of cows fed diets containing molasses adsorbed to soybean hull and wheat bran. The wheat bran diet resulted in significantly lower ruminal NH3-N concentrations than the control diet (P<0.05), however, no differences were observed between soybean hull and control diets (P>0.05). Furthermore, no significant differences (P>0.05) were observed in individual volatile fatty acid (VFA) (acetate, propionate, isobutyrate, butyrate, isovalerate, and valerate) concentrations or the acetate/propionate (A∶P) ratio between the control and treatment groups. However, the ruminal total VFA concentration in cows of the soybean hull molasses group did show a lower trend than that observed in the control group, whereas ruminal total VFA concentration in cows of the wheat bran molasses group had a tendency to increase. These results suggest that it would be efficient that feed soybean hull molasses to replace 15% of corn meal instead of 10% of what bran and 5% corn meal in dairy cattle. Molasses adsorbed to both wheat bran and soybean hull have the ability to maintain ruminal homeostasis, although wheat bran is a better adsorption substrate than soybean hull.

soybean molasses; dairy cows; ruminal environment

Zhou Chuan-she E-mail:zcs@isa.ac.cn Zhang Pei-hua E-mail:peiqin41-@163.com

2016-03-22接受日期：2016-08-20

國(guó)家自然科學(xué)基金面上項(xiàng)目(31372342)；中國(guó)科學(xué)院科技服務(wù)網(wǎng)絡(luò)計(jì)劃課題(KFJ-EW-STS-071)；中國(guó)科學(xué)院戰(zhàn)略性先導(dǎo)科技專項(xiàng)(XDA05020700)；中央駐湘科研機(jī)構(gòu)技術(shù)創(chuàng)新發(fā)展專項(xiàng)(2013TF3006)

韓奇鵬(1988-)，男，內(nèi)蒙古巴林右旗人，在讀碩士生，主要從事反芻動(dòng)物營(yíng)養(yǎng)與飼料研究。E-mail:18711069677@163.com

周傳社(1980-)，男，湖南汨羅人，研究員，博士，主要從事反芻動(dòng)物營(yíng)養(yǎng)研究。E-mail:zcs@isa.ac.cn 張佩華(1970-)，女，湖南汨羅人，副教授，博士，主要從事反芻動(dòng)物營(yíng)養(yǎng)與飼料研究。E-mail:peiqin41-@163.com

10.11829/j.issn.1001-0629.2016-0143

S823.9+1;S816.11

A

1001-0629(2016)12-2559-06*

韓奇鵬,張佩華,羅玲,揭紅東,周傳社,陳亮,孔志偉.不同來源吸附載體大豆糖蜜對(duì)奶牛瘤胃內(nèi)環(huán)境的影響.草業(yè)科學(xué),2016,33(12)：2559-2564.

Han Q P,Zhang P H,Luo L,Jie H D,Zhou C S,Chen L,Kong Z W.Effects of wheat bran and bean hull as adsorption substrates for soybean molasses on the ruminal environment of dairy cows.Pratacultural Science，2016,33(12)：2559-2564.