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上海地区奶牛非常规饲料的安全评估及应用
Safe Evaluation of Shanghai Unconventional Ingredients and Its Application
【作者】 范占炼;
【导师】 沈向真;
【作者基本信息】 南京农业大学 , 兽医, 2013, 博士
【摘要】 为合理开发和科学配置上海及周边地区非常规饲料资源,建立CPM配方软件数据库,大幅度降低饲料成本,避免人畜争粮,避免奶牛与其他单胃动物争夺饲料资源,我们重点对豆腐渣、酱油渣、老白酒糟等蛋白饲料和豆秸、稻草、羊草、小麦秸和青贮玉米等粗饲料进行系统分析,根据净碳水化合物净蛋白体系(CNCNP)理论,分析指标包括10个常规指标(干物质,粗蛋白,中性洗涤纤维,酸性洗涤纤维,木质素,粗脂肪,淀粉,糖,非纤维碳水化合物,灰分),6个能量指标(总消化养分,1倍消化能,1倍代谢能,3倍泌乳净能,3倍维持净能,3倍增重净能)和2个24h体外消化率指标(24h干物质体外消化率,24h中性洗涤纤维消化率),7种常量元素(钙,磷,镁,钾,硫,钠,氯)和5种微量元素(铁,锌,铜,锰,钼),3种蛋白组分(中性洗涤不溶蛋白,酸性洗涤不溶蛋白,溶解蛋白)等。经评估,豆腐渣、酱油渣、老白酒糟消化率高,能量高,可以作为优质蛋白饲料使用,以节省豆粕用量。豆秸、稻草、小麦秸和羊草成分相似,可以代替羊草。青贮玉米品质较国外低。对103种非常规饲料进行8种概略养分(水分,粗蛋白质,脂肪,灰分,NDF,ADF,钙,磷),部分品种进行真蛋白,糖,盐分,钾,中性洗涤不溶蛋白,酸性洗涤不溶蛋白分析,并建立相应数据库,为配合等营养日粮寻找原料提供科学依据。对1098个奶牛饲料样品AFB1检测,阳性率达到73.13%;国标超标率为1.64%,WHO超标率为3.83%,欧盟超标率15.85%。经评估,高危饲料产品为棉籽(光棉籽,毛棉籽,棉粕)及红薯秧捆;中度污染产品有:玉米DDGS,进口玉米DDGS,人工裹包发酵TMR,矮象草粉,烘干橙皮,光棉籽,棉饼,酵母培养物,白酒糟,膨化玉米尿素,复合糖蜜,玉米皮,真姬菇菌渣。污染较轻的有:新鲜TMR,大豆产品(大豆,膨化大豆,大豆皮和豆粕),麦类产品(大麦,大麦麸,大麦青贮,小麦,小麦麸,小麦青贮,小麦秸秆,啤酒糟,麦糟,麦乳精糟),玉米产品(玉米,压片玉米,玉米青贮),甜菜粕,干草(苜蓿草,羊草,野干草,牛毛草,澳大利亚燕麦草)。样品中全部符合欧盟标准的有:压片玉米,小麦,大麦,向日葵粕,黄酒糟,啤酒糟,麦糟,酵母饲料,豆粕,小麦麸,大麦麸,大豆皮,苹果渣,膨化大豆,大豆,澳大利亚燕麦草,羊草,苔草,野干草,牛毛草,青贮类产品(青贮大麦,青贮小麦,青贮稻草,青贮大米草),小麦秸秆,橙皮茶叶半干青贮,自动裹包发酵TMR。用ELISA对长征奶牛场部分原料的5种霉菌毒素(黄曲霉毒素B1,玉米赤霉烯酮,呕吐毒素,T-2毒素和烟曲霉毒素)分析表明,黄曲霉毒素B1为20ug/kg,不是本试验奶牛饲料中的主要霉菌毒素,不会引起牛奶中黄曲霉毒素M1的残留量超标;玉米赤霉烯酮100ug/kg,呕吐毒素500ug/kg,T-2毒素80ug/kg和烟曲霉毒素100ug/kg,在奶牛饲料中阳性率高,部分超标率高,超标严重。故日粮设计中试验组使用美国奥特奇霉菌毒素吸附剂”霉可吸”,观察其生产效果。试验表明在不影响泌乳奶牛乳脂率和乳蛋白率的前提下,霉菌毒素吸附剂”霉可吸”有提高奶牛产奶量的作用,整个试验期的平均产奶量与对照组比较提高了1.56kg。霉可吸可增强奶牛自身机体的免疫力,降低牛奶中的体细胞数,试验结束时霉可吸组比对照组体细胞多降低11万/ml,从而提高牛奶质量。牛奶中黄曲霉毒素M1的残留量未检出,检出限为0.008ug/L。总的来说,泌乳奶牛饲料中添加霉可吸能够提高奶牛场的经济效益。本项目用非常规饲料原料生产的奶牛全混合日粮(TMR),在国内首次采用超高压液相色谱-质谱/质谱(UHPLC-MS/MS),一次性能同时检测38种霉菌毒素,结果检出7种霉菌毒素,阳性检出率18.42%,分属8种霉菌属中的7种,说明了多霉菌毒素感染,也反映了感染的复杂性。各霉菌毒素浓度分别是:黄曲霉毒素G228.66±8.87ug/kg,呕吐毒素264.68±128.82ug/kg,HT-237.80±21.15ug/kg:伏马菌素B216.71±5.14ug/kg,神经毒素C0.47±0.13ug/kg,青霉酸56.80±5.20ug/kg;麦角新素(麦角酯醇)2.57±0.84ug/kg,但含量均在国家规定的限量范围内。此次没有检测到黄曲霉毒B1,但检测到黄曲霉毒素G2,提示我们光检测黄曲霉毒B1并不能真实反映饲料黄曲霉毒素的实际感染。牛奶黄曲霉毒素M181.75±87.28ng/kg也在允许范围内。说明饲喂非常规饲料配制的TMR生产的牛奶对人体是安全的。在后备牛日粮中,大米草代替青贮玉米作为粗料来源,用啤酒糟,低蛋白豆粕代替豆粕,用大麦代替玉米,在日粮营养水平相近的前提下,配制发酵TMR,饲喂6月龄的后备牛,其月增高、月增长和月增重、平均每日采食量分别增加了6.7%、7.7%、15.8%、13.9%,还能够大幅度降低后备奶牛的饲养成本,每吨饲料成本降低246.5元。这为人畜争粮、争地,消耗成本上涨,避免农产品价格过快上涨提供一个技术思路。泌乳中期荷斯坦奶牛以精料15%(1kg/d.head)小麦代替玉米,并添加木聚糖酶,加酶小麦组产量每头天增加鲜奶0.97kg,折成4%标准乳增加0.44kg,乳脂率增加0.2个百分点,乳蛋白减少0.08个百分点。对亚临床酮病发病率没有影响,粪便粘度正常。小麦和玉米有较好的饲料协作效应。在泌乳高峰期,利用CPM电脑软件,应用非常规蛋白饲料:新鲜豆腐渣、米酒糟和低盐酱油渣、双低菜粕和浮性缓释尿素膨化玉米颗粒;能量饲料压片小麦、粗料:青贮大麦、发酵橙皮茶叶渣饲料、干稻草代替传统的蛋白饲料豆粕、膨化大豆和DDGS;部分能量饲料玉米和部分粗料进口苜蓿草、燕麦草和羊草,使日粮成本下降18.45元,原料进口依存度从41.02%进一步减少到15.33%,日单产增加0.81kg,乳脂肪,乳蛋白,乳糖,固形物,乳尿素氮变化不大,体细胞下降17.6万/ml,血中酮体BHBA0.85mmol/L、 AOAC35.80mg/100ml,属正常水平,变化不大,总抗氧化能力5.82u/ml和过氧化氢酶活性0.63u/ml,比对照组高,尤其是过氧化氢酶活性达到显著差异。SOD95.2u/ml、GSH-PX184.17u/ml,匕对照组稍低,但差异不显著,脂质过氧化物MDA0.70nmol/ml和NO14.74nmol/ml较低。发酵橙皮茶叶渣饲料可能对降低体细胞和提高机体抗氧化能力有一定的好处。
【Abstract】 In order to develop Shanghai new unconventional ingredient and build CPM formulation software feed database, we sampled tofu residue, soy sauce pulp, rice distillers grains,soy straw, rice straw, corn silage,wheat straw and wild rye grass to AmericanDairyone Forage Testing Laboratory to analysis feed composition and nutrition. Analysis index included10conventional index (DM, CP, EE, NDF, ADF, Starch, Lignin, NFC, ESC, Ash),6energy index(TDN1X, DE1X, ME1X, NEL3X, NEM3X, NEG3X),2digestibility(24hIVTD,24hNDFD),7macroelements (DCAD, Ca, P, Mg, K, S, Na, Cl),5microelements(Fe, Zn, Cu, Mn, MO),3N fraction (NDICP, ADICP, SP).Analysis showed tofu residue, soy sauce pulp, and rice distillers grains had high digestibility and energy, can be used as quality protein resource and can substitute for soybean meal;soy straw, rice straw, wheat straw had the similar composition of wild rye grass,and can replace each other.Other102unconventional ingredients were sent to Shanghai Institute of Dairy Cattle to test DM,CP, EE, NDF, ADF,NFC,Ca,P and Ash.Some also were tested true protein, suger,salt,K,NDICP and ADICP.Feed database were set up for future formulation.Survey of AFB1content in1098dairy cattle ingredientsample showed occurency rate was73.13%;1.64%feeds were was above the PRC action level of50μg/kg,3.83%feeds were above the WHO action level of15μg/kg,15.85%feeds were above the EU action level of5μg/kg.High level AFB1was founded in cotton seed and cotton seed meal.We analysed AFB1,ZON, DON, T-2, Fumonisin level in Some ingredients from trial dairy cattle farm. Analysis show AFB1content is20ug/kg, isn’t major contaminated mycotoxin,can’t cause AFM1is hgher than regulation upper limit.Other mycotoxins content is ZON100ug/kg, DON500ug/kg,T-280ug/kg, Fumonisin100ug/kg. These mycotoxins contamination is serious.So we addited mycotoxin binder "mycosorber" from ALLTECH CO.,Ltd. to experiment group to oberserv it’s effect. Experiment showed mycobinder increased milk yield1.56kg/d.head more, reduced SCC110,000/ml,no effect on milk protein and butter content.AFM1is0.008ug/L,much lower than regulation upper limit0.5ug/L. Mycotoxin binder has good response on milking performance.38type mycotoxins in Shanghai dairy cattle unconventional TMR (Total Mixed Ration) were tested by using UHPLC/UHPLC-MS/MS. This method can test38type mycotoxins in one time.Analysis showed7type mycotoxins were positive.This show mycotoxin comtamination is very complicated.Positive rate is18.42%.Each mycotoxin content is AFG2,28.66±8.87ug/kg, DON264.68±128.82ug/kg, HT-237.80±21.15ug/kg, Fumonisin B216.71±5.14ug/kg, Roquefortine C0.47±0.13ug/kg, Penicillic Acid56.80±5.20ug/kg, Ergometrine2.57±0.84ug/kg,respectively.These mycotoxins content were not too much highe.Feed Aflatoxin has many subtype,so we must test AFB1, AFB2, AFG1, AFG2to know true Aflatoxin contamination,Milk Aflatoxin M1from cow feeded unconventional TMR is81.75±87.28ng/kg, is lowerer than upper limit of P.R.C. Regulation。In6month old heifer fermented TMR,We use invasive Spartina alterniflora to replace corn silage as forage,use soybean isolated protein byproduc,wet brewer grain to substitute for soybean meal,use barley to substitute for corn.Ration nutrition is the same as control ration. Experiment shows unconventional ration improve heifer height at6.7%more,gain at15.8%more,length at7.7%more and body weight at13.9%more,reduce ration cost246.5yuan RMB/t.This model is one of solution for crisis of feed,land and rapid cost increase.In middle lactation concentration,15%ground corn was substituted for wheat with xylanase.Wheat based group showed0.97kg/d.head more milk yield(equal to0.44kg4%FCM),0.2%more butter level,0.08%less protein level,no effect on ketosis rate and feces viscosity,more return3.15yuan RMB/d.head. Wheat and corn have good cooperation response.In peak lactation with help of CPM Dairy Ration Analyzer V3.0.8,without big nutrition chang,we used unconventional protein feed resource such as fresh tofu residue, rice distillers grains, low salt soysauce pulp,canola meal,slow realeased urea expand corn pellet to replace soybean meal and extrution soy;use steamed flaked wheat to substitute for Corn,use barley silage,fermented citrus pulp tea mix,rice straw to partly substitute for imported aflafal hay, oat hay and deyeuxia angustifolia. Unconventional TMR cost is18.45yuan/d.head RMB lower than conventional TMR cost, used less import feed from41.02%to15.33%, increased milk production0.81kg/d.head. Milk composition including butter,protein,lactose,solide,MUN has small change.Milk SCC has reduction.BHBA level is 0.85mmol/L,is the same as control group.ACAC is35.80mg/100ml, has small change.Ketone body content is normal in both two group.Compare to control group, T-AOC(5.82u/ml) and CAT(0.63u/ml) is higher.CAT level was different significantly(P<0.05).Level of SOD,GSH-PX is95.2u/ml,184.17u/ml respectively, lower than than of control group.Content of MDA, NO in serum is0.70nmol/ml,14.74nmol/ml respectively, lower than that of control group. Higher antioxidation capacity might be caused by fermented citrus pulp tea mix riched in antioxydative ingredients.
【Key words】 Mycotoxin; Unconventional ingredient; TMR; Dairy cattle; UHPLC/UHPLC-MS/MS; Milk; AFM1;