殷雨洋,劉玉潔,張瑞陽,朱偉云,毛勝勇*
(1.南京農(nóng)業(yè)大學(xué)動(dòng)物科技學(xué)院,江蘇 南京 210095;2.湖州市農(nóng)業(yè)科學(xué)研究院湖羊研究所,浙江 湖州 313000)
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高精料日糧下添加阿卡波糖對(duì)奶牛瘤胃及后腸發(fā)酵的影響研究
殷雨洋1,2,劉玉潔1,張瑞陽1,朱偉云1,毛勝勇1*
(1.南京農(nóng)業(yè)大學(xué)動(dòng)物科技學(xué)院,江蘇 南京 210095;2.湖州市農(nóng)業(yè)科學(xué)研究院湖羊研究所,浙江 湖州 313000)
本試驗(yàn)旨在探討高精料日糧下添加阿卡波糖對(duì)奶牛瘤胃和后腸發(fā)酵的影響。試驗(yàn)選用3頭干奶期荷斯坦奶牛,采用3×3拉丁方試驗(yàn)設(shè)計(jì),阿卡波糖添加劑量為0,0.5和1.0 g/d,試驗(yàn)分3期進(jìn)行,每期21 d。結(jié)果表明,與對(duì)照組比較,添加阿卡波糖顯著降低了奶牛瘤胃液中丙酸濃度(P<0.05),提高了乙丙比(P<0.05),但對(duì)瘤胃pH值、乳酸、乙酸、異丁酸、丁酸、異戊酸、戊酸、總揮發(fā)性脂肪酸和氨氮濃度無顯著影響(P>0.05);與對(duì)照組比較,添加阿卡波糖顯著降低了糞便pH值和氨氮濃度(P<0.05),提高了乳酸、丁酸和異戊酸濃度(P<0.05),但對(duì)乙酸、丙酸、異丁酸、戊酸、總揮發(fā)性脂肪酸和乙丙比無顯著影響(P>0.05)。結(jié)果說明,高精料日糧下長期添加阿卡波糖雖可影響瘤胃液中個(gè)別揮發(fā)性脂肪酸的濃度,但對(duì)瘤胃整體發(fā)酵和瘤胃pH值無顯著影響,此外,添加阿卡波糖可增加后腸發(fā)酵,并可能對(duì)后腸健康帶來潛在危害。
阿卡波糖;奶牛;瘤胃;后腸;揮發(fā)性脂肪酸
在現(xiàn)代奶牛生產(chǎn)中,為提高奶牛產(chǎn)奶性能和經(jīng)濟(jì)效益,生產(chǎn)者常在奶牛日糧中使用高比例精料,但大量使用精料易導(dǎo)致瘤胃內(nèi)揮發(fā)性脂肪酸(volatile fatty acid,VFA)和乳酸等累積,pH值快速下降,結(jié)果誘發(fā)瘤胃酸中毒[1]。瘤胃酸中毒可分為急性酸中毒(acute ruminal acidosis,ARA)和亞急性酸中毒(subacute ruminal acidosis,SARA),其中SARA在生產(chǎn)中較為常見。有研究顯示,即使在美國管理較嚴(yán)格的奶牛場,早期泌乳?;?SARA的比例約為19%,中期泌乳牛SARA發(fā)病率甚至高達(dá)26%[2]。由于該病臨床癥狀不明,同時(shí)該病常造成其他繼發(fā)性疾病如蹄葉炎、腹瀉和肝膿腫等的發(fā)生[3-8]。因此,該病已成為影響奶牛生產(chǎn)效益的主要疾病之一。近年來,許多營養(yǎng)措施被用于防治奶牛瘤胃酸中毒發(fā)生,具體包括:增加物理有效纖維含量;降低飼料中谷物類飼料的比例;使用緩沖劑、添加酵母培養(yǎng)物和應(yīng)用莫能霉素[9-10]。這些方式在生產(chǎn)中雖取得了較顯著的效果;但也表現(xiàn)出一些負(fù)面效應(yīng),如提高物理有效纖維的用量可導(dǎo)致日糧能量濃度下降,最終導(dǎo)致奶牛產(chǎn)奶性能下降;而緩沖劑存在降低瘤胃pH的作用時(shí)間短暫的問題;而應(yīng)用莫能菌素雖可預(yù)防SARA的發(fā)生,但也帶來瘤胃微生物耐藥性增加和瘤胃纖維降解率下降的風(fēng)險(xiǎn)[11]。因此,尋找新的酸中毒預(yù)防措施,已成為當(dāng)前反芻動(dòng)物營養(yǎng)研究領(lǐng)域的一個(gè)熱點(diǎn)問題。
近年來,一些學(xué)者發(fā)現(xiàn),使用阿卡波糖等α-葡糖苷酶和α-淀粉酶抑制劑可有效控制和緩解奶牛急性瘤胃酸中毒發(fā)生[12];同時(shí)可提高采食量和乳脂校正奶產(chǎn)量[13]。我們?cè)谝郧暗捏w外實(shí)驗(yàn)中也發(fā)現(xiàn),阿卡波糖可顯著提高發(fā)酵體系中瘤胃pH值[14],但長期飼喂阿卡波糖的效果如何,并不清楚。此外,有報(bào)道顯示[15],在治療人糖尿病過程中,阿卡波糖雖可有效控制人小腸內(nèi)碳水化合物的降解速度,但同時(shí)也提高了后腸中淀粉數(shù)量,進(jìn)而促進(jìn)了微生物發(fā)酵;由于后腸健康對(duì)奶牛生產(chǎn)十分關(guān)鍵,阿卡波糖在影響瘤胃碳水化合物的降解的同時(shí),是否會(huì)增加奶牛后腸消化道負(fù)擔(dān),并不清楚。由此,本試驗(yàn)基于以上進(jìn)展,研究了在高精料日糧下添加阿卡波糖對(duì)奶牛瘤胃及后腸發(fā)酵的影響,擬進(jìn)一步明確阿卡波糖對(duì)奶牛瘤胃與后腸發(fā)酵的實(shí)際影響效果。
實(shí)驗(yàn)于2013年11月到2014年1月在南京農(nóng)業(yè)大學(xué)江浦實(shí)驗(yàn)基地進(jìn)行。
1.1實(shí)驗(yàn)動(dòng)物及日糧
本試驗(yàn)選用3頭干奶期裝有永久性瘤胃瘺管的健康多胎奶牛為試驗(yàn)動(dòng)物,單槽飼喂,自由飲水,每天8:00和16:00進(jìn)行飼喂,體重為(590±48) kg?;A(chǔ)飼糧精粗比為65∶35,滿足中國農(nóng)業(yè)行業(yè)標(biāo)準(zhǔn)(NY/T34-2004)推薦的營養(yǎng)需求,日糧配方及營養(yǎng)水平見表1。
1.2試驗(yàn)試劑
阿卡波糖(Acarbose)為美國Bayer公司生產(chǎn),用于治療Ⅱ型糖尿病的藥物。
1.3試驗(yàn)設(shè)計(jì)
試驗(yàn)采用3×3拉丁方設(shè)計(jì),分3期進(jìn)行,每期21 d。在正式試驗(yàn)前預(yù)飼10 d。正式試驗(yàn)前1 d對(duì)奶牛稱重。阿卡波糖添加劑量為0,0.5和1.0 g/d。
表1 日糧配方及營養(yǎng)水平Table 1 Ingredient and nutrient levels of the diets
[1]每kg添加劑包括 The premix contains per kilogram: Fe 5500 mg, Cu 4080 mg, Zn 17500 mg, Mn 4980 mg, Se 110 mg, I 180 mg, Co 88.5 mg, VA>2000000 IU, VD3600000 IU, VE 10800 mg.
1.4樣品采集與分析
1.4.1干物質(zhì)采食量的測定在每期的第18,19和20天,分別對(duì)飼喂量和剩余料量進(jìn)行稱重,取樣測定干物質(zhì)重,計(jì)算干物質(zhì)采食量。
1.4.2瘤胃和后腸發(fā)酵參數(shù)的測定在每期的第18,19和20天,分別采集飼喂阿卡波糖后0和2 h瘤胃內(nèi)容物,并在晨飼前直腸采集300 g糞便;在每期第21天采集飼喂阿卡波糖后0,2,4,6,8,10和12 h的瘤胃內(nèi)容物。所采集瘤胃內(nèi)容物經(jīng)4層紗布過濾獲得瘤胃液,立即測定瘤胃液和糞便pH值。取200 g糞便與200 mL去離子水混合均勻后,15000 r/min下離心,取上清液于-20℃凍存?zhèn)溆?。采用比色法測定瘤胃液和糞便中乳酸[16]和氨氮濃度[17],采用氣相色譜法 (GC-14B氣相色譜儀,日本島津,柱溫110℃,氣化室溫度180℃,檢測室溫度180℃) 測定揮發(fā)性脂肪酸 (volatile fatty acid,VFA) 濃度[18]。
1.5數(shù)據(jù)處理
試驗(yàn)各期數(shù)據(jù)采用SPSS (17.0)軟件中GLM模型中Multivariate程序進(jìn)行主效應(yīng)分析,以牛體重為協(xié)變量,當(dāng)阿卡波糖的處理效應(yīng)達(dá)顯著水平時(shí),對(duì)阿卡波糖各處理間的差異進(jìn)一步進(jìn)行多重比較,多重比較采用LSD法。顯著性水平為P<0.05。
圖1 添加阿卡波糖對(duì)奶牛干物質(zhì)采食量的影響Fig.1 Effects of acarbose addition on the dry matter intake of dairy cows
2.1添加阿卡波糖對(duì)奶牛干物質(zhì)采食量的影響
圖1為添加阿卡波糖對(duì)奶牛干物質(zhì)采食量的影響。與對(duì)照組比較,添加阿卡波糖有提高奶牛干物質(zhì)采食量的趨勢(P<0.1)。
2.2添加阿卡波糖對(duì)奶牛瘤胃發(fā)酵的影響
由表2可見,較對(duì)照組相比,添加阿卡波糖顯著降低了瘤胃液中丙酸含量(P<0.05),提高了乙丙比(P<0.05),但對(duì)瘤胃pH、乳酸、乙酸、異丁酸、丁酸、異戊酸、戊酸、總揮發(fā)性脂肪酸(total volatile fatty acid, TVFA)和氨氮濃度無顯著影響(P>0.05)。飼
表2 添加阿卡波糖對(duì)瘤胃微生物發(fā)酵的影響Table 2 Effects of acarbose addition on the fermentation in rumen of dairy cows
SEM, standard error of the mean; A/P, ratio of acetate to propionate; TVFA, total volatile fatty acid. 不同字母表示差異顯著(P<0.05)。Values with different letters mean significantly difference (P<0.05). 下同。The same below.
喂期對(duì)瘤胃pH值、乙酸、丙酸、異丁酸、丁酸和TVFA濃度及乙丙比有顯著影響(P>0.05);在對(duì)瘤胃pH值、丙酸、丁酸、TVFA、氨氮濃度和乙丙比的影響上,飼喂期和阿卡波糖劑量之間有顯著的互作效應(yīng)(P<0.05)。
2.3添加阿卡波糖對(duì)奶牛瘤胃發(fā)酵指標(biāo)動(dòng)態(tài)變化的影響
較對(duì)照組相比,添加阿卡波糖對(duì)瘤胃pH的動(dòng)態(tài)變化無顯著影響(P>0.05),但在飼喂后0~4 h期間,添加阿卡波糖可減緩瘤胃pH值下降速率,提高最低瘤胃pH值;添加阿卡波糖對(duì)乙酸、丙酸和丁酸濃度的動(dòng)態(tài)變化無顯著影響(P>0.05)。與對(duì)照組相比,添加阿卡波糖對(duì)總揮發(fā)性脂肪酸濃度、乙丙比和乳酸濃度的動(dòng)態(tài)變化無顯著影響(P>0.05)。此外,添加阿卡波糖顯著降低了瘤胃氨氮濃度(P<0.05)(圖2)。
圖2 添加阿卡波糖對(duì)奶牛瘤胃pH值、VFA、乳酸、氨氮濃度及乙丙比動(dòng)態(tài)變化的影響Fig.2 Effects of acarbose addition on changes in the ruminal pH, the concentrations of VFA, lactate and NH3-N, and the ratio of acetate to propionate
2.4添加阿卡波糖對(duì)奶牛后腸發(fā)酵的影響
與對(duì)照組相比,添加阿卡波糖顯著降低了后腸pH值和氨氮水平(P<0.05),顯著提高了乳酸、丁酸和異戊酸濃度(P<0.05),但對(duì)乙酸、丙酸、異丁酸、戊酸、TVFA濃度及乙丙比沒有顯著(P>0.05)影響。飼喂期對(duì)瘤胃pH值、乳酸、丙酸和丁酸濃度有顯著影響(P<0.05),阿卡波糖劑量和飼喂周期對(duì)pH值、丁酸和異戊酸濃度亦有顯著的互作效應(yīng)(P<0.05)(表3)。
表3 添加阿卡波糖對(duì)奶牛后腸發(fā)酵的影響Table 3 Effects of acarbose addition on the hindgut fermentation in dairy cows
隨著我國奶牛飼養(yǎng)規(guī)模的不斷增大與集約化程度的不斷提高,以及生產(chǎn)者對(duì)經(jīng)濟(jì)效益的過度追求,因大量飼喂高精料日糧而誘發(fā)的瘤胃健康紊亂問題已嚴(yán)重影響到我國一些牧場的正常生產(chǎn)。研究發(fā)現(xiàn)[19-20],高精料日糧易導(dǎo)致瘤胃pH值過低,進(jìn)而引發(fā)瘤胃內(nèi)代謝紊亂。因此,如何有效提高瘤胃pH值,對(duì)于防止瘤胃過酸帶來的健康隱患意義巨大。Speight和Harmon[21]體外研究發(fā)現(xiàn),添加阿卡波糖能有效防止瘤胃pH下降、TVFA和乳酸累積,進(jìn)而有效控制和緩解急性瘤胃酸中毒的發(fā)生。Speight等[22]報(bào)道,阿卡波糖可能通過降低瘤胃內(nèi)淀粉的降解速度來起到預(yù)防酸中毒的作用。與上述報(bào)道不一致,本試驗(yàn)結(jié)果表明,添加阿卡波糖雖可顯著降低瘤胃中丙酸含量,提高乙丙比,但對(duì)瘤胃pH值、氨氮、乳酸和總揮發(fā)性脂肪酸濃度無顯著影響;此外,對(duì)瘤胃pH值動(dòng)態(tài)變化數(shù)值的統(tǒng)計(jì)結(jié)果顯示,阿卡波糖對(duì)瘤胃pH值也無顯著效果。該結(jié)果暗示,長期飼喂阿卡波糖不能有效提高瘤胃pH,暗示其在預(yù)防奶牛瘤胃亞急性酸中毒方面的作用效果有限。本研究表明,盡管阿卡波糖對(duì)瘤胃和后腸大多指標(biāo)影響不顯著,但飼喂期對(duì)多項(xiàng)指標(biāo)有顯著影響;而且在多項(xiàng)指標(biāo)上,飼喂期和阿卡波糖劑量之間有顯著的互作效應(yīng)。我們推測,出現(xiàn)上述結(jié)果的根本原因可能在于阿卡波糖在影響微生物發(fā)酵方面的短暫效應(yīng),即動(dòng)物初次使用阿卡波糖可能存在一定效果,但長期使用則效果減弱或消失,原因可能在于瘤胃微生物種類十分豐富,長期添加阿卡波糖使部分瘤胃微生物對(duì)其產(chǎn)生了適應(yīng),進(jìn)而使其生物學(xué)效應(yīng)下降,但該推測仍需試驗(yàn)驗(yàn)證。
近年來,對(duì)阿卡波糖的相關(guān)研究主要關(guān)注于其對(duì)瘤胃發(fā)酵的影響,對(duì)后腸消化有何具體影響,仍不清楚。Nordlund等[3]報(bào)道,飼喂高精料日糧除可顯著影響瘤胃健康外,還可促進(jìn)后腸發(fā)酵,導(dǎo)致后腸食糜pH值降低,進(jìn)而可能引發(fā)氣泡糞和腹瀉產(chǎn)生。本試驗(yàn)結(jié)果顯示,飼喂阿卡波糖顯著降低了后腸pH值和氨氮濃度,提高了乳酸、乙酸、丁酸和異戊酸的濃度。試驗(yàn)結(jié)果說明,應(yīng)用阿卡波糖加重了奶牛后腸道的消化負(fù)擔(dān),可能對(duì)奶牛后腸道健康造成危害。其原因可能與阿卡波糖抑制了奶牛小腸淀粉酶活性,導(dǎo)致奶牛小腸淀粉消化率下降,最終進(jìn)入奶牛后腸中的淀粉數(shù)量增加,進(jìn)而使奶牛后腸微生物發(fā)酵增強(qiáng),導(dǎo)致部分VFA濃度增加,pH值下降。
綜上所述,奶牛日糧中長期添加阿卡波糖對(duì)瘤胃pH值、乳酸和TVFA濃度未產(chǎn)生顯著影響,但顯著降低了后腸pH值,提高了乳酸含量。因此,高精料日糧下長期添加阿卡波糖不能改善奶牛瘤胃健康,并可能增加奶牛后腸發(fā)酵,影響奶牛后腸健康。
References:
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*The effect of acarbose addition on rumen and hindgut fermentation in dairy cows fed high-concentrate diets
YIN Yu-Yang1,2, LIU Yu-Jie1, ZHANG Rui-Yang1, ZHU Wei-Yun1, MAO Sheng-Yong1*
1.TheCollegeofAnimalScienceandTechnology,NanjingAgriculturalUniversity,Nanjing210095,China;2.HuzhouAcademyofAgriculturalSciences,Huzhou313000,China
In this study, three cannulated Holstein cows (590±48 kg liveweight) were used to investigate the effect of acarbose (glucosidase inhibitor) addition (0, 0.5 and 1.0 g/d) on ruminal and hindgut fermentation characteristics in a 3×3 Latin square design. The results showed that acarbose addition decreased the propionate concentration in rumen fluid (P<0.05), increased the ratio of acetate to propionate (P<0.05) but did not affect the concentrations of lactate, acetate, isobutyrate, butyrate, isovalerate, valerate, total volatile fatty acid, ammonia-nitrogen, and pH value compared with the control (P>0.05). Acarbose addition decreased the pH value and ammonia-nitrogen levels (P<0.05) and increased the concentrations of lactate, butyrate, isovalerate (P<0.05) in feces, but no differences were observed in the concentration of acetate, propionate, isobutyrate, valerate, total volatile fatty acid and the ratio of acetate to propionate among the three groups. In general, our data revealed that acarbose addition may not be useful for preventing the occurrence of ruminal acidosis in the long term. Additionally, acarbose addition may increase hindgut fermentation and produce detrimental effects on the hindgut health of dairy cattle.
acarbose; diary cows; rumen; hindgut; volatile fatty acid
10.11686/cyxb2015431
http://cyxb.lzu.edu.cn
2015-09-09;改回日期:2015-11-09
國家“973”奶業(yè)項(xiàng)目(2011CB100801)資助。
殷雨洋(1987-),男,安徽阜陽人,助理畜牧師,碩士。E-mail: yinyuyang@163.com
Corresponding author. E-mail: maoshengyong@163.com
殷雨洋,劉玉潔,張瑞陽,朱偉云,毛勝勇. 高精料日糧下添加阿卡波糖對(duì)奶牛瘤胃及后腸發(fā)酵的影響研究. 草業(yè)學(xué)報(bào), 2016, 25(6): 198-203.
YIN Yu-Yang, LIU Yu-Jie, ZHANG Rui-Yang, ZHU Wei-Yun, MAO Sheng-Yong. The effect of acarbose addition on rumen and hindgut fermentation in dairy cows fed high-concentrate diets. Acta Prataculturae Sinica, 2016, 25(6): 198-203.