CN114158655B - L-苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法 - Google Patents
L-苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法 Download PDFInfo
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- CN114158655B CN114158655B CN202111552376.XA CN202111552376A CN114158655B CN 114158655 B CN114158655 B CN 114158655B CN 202111552376 A CN202111552376 A CN 202111552376A CN 114158655 B CN114158655 B CN 114158655B
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Abstract
本发明属于动物营养技术领域，具体涉及L‑苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法。本发明提供L‑苹果酸在改善猪肉品质中的应用。本发明所述L‑苹果酸能够在保育期通过干预猪骨骼肌的肌纤维类型转化，达到提高猪骨骼肌氧化型肌纤维的比例的目的，进而达到改善育肥猪肉品质的效果。
Description
技术领域
本发明属于动物营养技术领域，具体涉及L-苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法。
背景技术
2015年以来，我国畜禽肉总产量达到创纪录的8625万吨，随后进入平台期，基本维持在8500万吨左右，人均猪肉占有量超过了中国居民膳食指南推荐的每天50～75g畜禽肉摄入量。在生猪产业快速发展的过程中，由于对猪的生长速度和瘦肉率的过分追求，导致猪肉系水力降低，风味变差，劣质肉时有发生，猪肉品质和消费者预期仍然存在较大的差距。肉色、嫩度、风味、多汁性、系水力、肌内脂肪含量等是评价猪肉品质的关键指标。
L-苹果酸是三羧酸循环的中间代谢产物，易于吸收，通过细胞膜进入线粒体内，以苹果酸-草酰乙酸穿梭系统的形式参与线粒体能量代谢，具有调控肌肉能量代谢、推动肌纤维类型转化的潜力。目前的研究发现，一水肌酸和L-苹果酸组成的混合酸可以增加育肥猪的眼肌面积；在母猪妊娠后期利用有机酸(富马酸、柠檬酸和L-苹果酸)和中链脂肪酸(辛酸和癸酸)的混合物可以提高哺乳仔猪的平均日增重。然而，利用L-苹果酸改善猪肉品质的研究还未见报道。
发明内容
本发明的目的在于提供L-苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法。利用本发明所述的L-苹果酸能够有效改善猪肉品质。
为了实现上述目的，本发明提供如下技术方案：
本发明提供了L-苹果酸在改善猪肉品质中的应用。
本发明提供了一种改善猪肉品质的混合物，所述混合物包括L-苹果酸。
优选的，所述混合物还包括载体；所述载体包括蒙脱石和/或沸石粉。
优选的，所述L-苹果酸和载体的质量比为(2～3)：(7～8)。
本发明提供了一种改善猪肉品质的饲料，所述饲料包括上述技术方案所述的混合物和基础日粮。
优选的，所述混合物与基础日粮的质量比为(0.5～1)：(99～99.5)。
优选的，所述基础日粮包括以下质量份的组分：玉米62.00～63.45份、豆粕9.65～11.00份、膨化全脂大豆9.10～10.45份、鱼粉3.80～4.22份、乳清粉5.76～6.21份、大豆浓缩蛋白1.90～2.10份、豆油1.72～1.76份、磷酸氢钙0.63～0.66份、石粉1.04～1.12份、食盐0.28～0.31份、L-赖氨酸盐酸盐0.58～0.61份、DL-蛋氨酸0.19～0.21份、L-苏氨酸0.22～0.25份、L-色氨酸0.04～0.07份、植酸酶0.01份和预混料0.50份。
本发明提供了上述技术方案所述混合物或上述技术方案所述饲料在改善猪肉品质中的应用。
优选的，所述猪肉品质为育肥猪的猪肉品质。
本发明提供了一种改善猪肉品质的方法，包括以下步骤：
在猪保育期饲喂上述技术方案所述的饲料。
有益效果：
本发明提供L-苹果酸在改善猪肉品质中的应用。本发明所述L-苹果酸能够在保育期通过干预猪骨骼肌的肌纤维类型转化，达到提高猪骨骼肌氧化型肌纤维的比例的目的，进而达到改善育肥猪肉品质的效果。具体实施例结果表明：利用本发明所述L-苹果酸饲喂猪后能够显著增加保育猪骨骼肌的氧化型肌纤维比例。
而且，本发明所述L-苹果酸对猪生长育肥期的生长性能无不良影响，对育肥猪的胴体性状无不良影响，提高屠宰后24h的pH值，降低滴水损失，可见，L-苹果酸对猪肉品质有很好的改善作用。本发明在窗口期内进行营养干预，通过增加氧化型肌纤维的比例改善猪肉品质是促进猪肉优质生产的一个重要策略，为现有技术提高猪肉品质提供了依据。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例中所需要使用的附图作简单地介绍。
图1为不同处理对保育猪的肌纤维类型组成的影响，其中A为琥珀酸脱氢酶染色结果图，B为琥珀酸脱氢酶阳性的肌纤维的比例图。
具体实施方式
如无特殊要求，本发明所述组分均为本领域技术人员常规购买所得即可。
本发明提供了L-苹果酸在改善猪肉品质中的应用。本发明所述L-苹果酸能够在保育期促进猪骨骼肌的肌纤维类型转化，达到提高猪骨骼肌氧化型肌纤维的比例的目的，进而达到改善育肥猪肉品质的效果。本发明对所述L-苹果酸的来源没有特殊的限定，采用本领域技术人员常规购买所得即可；在本发明具体实施例中，所述L-苹果酸优选购自安徽雪郎生物科技股份有限公司，所述L-苹果酸的纯度优选为≥70.0％。
本发明提供了一种改善猪肉品质的混合物，所述混合物包括L-苹果酸。在本发明中，所述混合物还优选包括载体；所述载体优选包括蒙脱石和/或沸石粉，更优选包括蒙脱石和沸石粉。在本发明中，所述L-苹果酸和载体的质量比优选为(2～3)：(7～8)，更优选为(2.4～2.7)：(7.3～7.6)；当所述载体包括蒙脱石和沸石粉时，所述蒙脱石和沸石粉的质量比优选为(3.5～4)：(3.5～4)，更优选为(3.6～3.8)：(3.9～4)。本发明对所述蒙脱石和沸石粉的来源没有特殊限定，采用本领域技术人员常规购买所得即可；在本发明具体实施例中，所述蒙脱石和沸石粉均优选购自北京同力兴科农业科技有限公司，所述蒙脱石和沸石粉的纯度均优选为98％。
以质量份计，本发明所述混合物优选包括2～3份L-苹果酸和7～8份载体。本发明所述混合物更优选为2.4～2.7份L-苹果酸。在本发明中，所述L-苹果酸能够在保育期促进猪骨骼肌的肌纤维类型转化，达到提高猪骨骼肌氧化型肌纤维的比例的目的，进而达到改善育肥猪肉品质的效果。
以L-苹果酸的质量份为基准，本发明所述混合物优选包括7～8份载体，更优选为7.3～7.6份。在本发明中，所述载体优选包括包括蒙脱石和/或沸石粉，更优选包括蒙脱石和沸石粉；当所述载体包括蒙脱石和沸石粉时，所述蒙脱石优选为3.5～4份，更优选为3.75～4份，沸石粉优选为3.5～4份，更优选为3.75～4份。本发明所述混合物通过该载体能够进入猪的体内，进而保证L-苹果酸的作用。
本发明提供了一种改善猪肉品质的饲料，所述饲料包括上述技术方案所述的混合物和基础日粮。在本发明中，所述混合物与基础日粮的质量比为(0.5～1)：(99～99.5)，更优选为(0.6～0.8)：(99.2～99.4)，以质量百分含量计，所述混合物的含量优选为0.5％～1％，更优选为0.6％～0.8％，基础日粮的含量优选99％～99.5％，更优选为99.2％～99.4％。
在本发明中，所述基础日粮优选包括以下质量份的组分：玉米62.00～63.45份、豆粕9.65～11.00份、膨化全脂大豆9.10～10.45份、鱼粉3.80～4.22份、乳清粉5.76～6.21份、大豆浓缩蛋白1.90～2.10份、豆油1.72～1.76份、磷酸氢钙0.63～0.66份、石粉1.04～1.12份、食盐0.28～0.31份、L-赖氨酸盐酸盐0.58～0.61份、DL-蛋氨酸0.19～0.21份、L-苏氨酸0.22～0.25份、L-色氨酸0.04～0.07份、植酸酶0.01份和预混料0.50份，更优选为玉米62.5～63.45份、豆粕9.90～10.5份、膨化全脂大豆9.50～10.30份、鱼粉3.90～4.10份、乳清粉5.90～6.10份、大豆浓缩蛋白1.95～2.00份、豆油1.73～1.75份、磷酸氢钙0.64～0.65份、石粉1.06～1.10份、食盐0.29～0.30份、L-赖氨酸盐酸盐0.59～0.60份、DL-蛋氨酸0.20份、L-苏氨酸0.23～0.24份、L-色氨酸0.05～0.06份、植酸酶0.01份和预混料0.50份。
在本发明中，以质量百分含量计，所述基础日粮优选包括以下质量份的组分：玉米62.00％～63.45％、豆粕9.65％～11.00％、膨化全脂大豆9.10％～10.45％、鱼粉3.80％～4.22％、乳清粉5.76％～6.21％、大豆浓缩蛋白1.90％～2.10％、豆油1.72％～1.76％、磷酸氢钙0.63％～0.66％、石粉1.04％～1.12％、食盐0.28％～0.31％、L-赖氨酸盐酸盐0.58％～0.61％、DL-蛋氨酸0.19％～0.21％、L-苏氨酸0.22％～0.25％、L-色氨酸0.04％～0.07％、植酸酶0.01％和预混料0.50％，更优选为玉米62.5％～63.45％、豆粕9.90％～10.5％、膨化全脂大豆9.50％～10.30％、鱼粉3.90％～4.10％、乳清粉5.90％～6.10％、大豆浓缩蛋白1.95％～2.00％、豆油1.73％～1.75％、磷酸氢钙0.64％～0.65％、石粉1.06％～1.10％、食盐0.29％～0.30％、L-赖氨酸盐酸盐0.59％～0.60％、DL-蛋氨酸0.20％、L-苏氨酸0.23％～0.24％、L-色氨酸0.05％～0.06％、植酸酶0.01％和预混料0.50％。本发明所述基础日粮能够满足猪生长过程中所需营养，满足NRC(2012)中规定的猪的营养需要。
在本发明中，以1kg饲料计，所述预混料优选包括维生素A 10000IU、维生素D31500IU、维生素E 30IU、维生素K3 2.5mg、维生素B1 1.5mg、维生素B2 10mg、维生素B610mg、维生素B120.05mg、叶酸1mg、生物素0.5mg、烟酸30mg、泛酸20mg、铜20mg、铁100mg、锌110mg、锰40mg、硒0.3mg和碘0.54mg。
本发明提供了上述技术方案所述混合物或上述技术方案所述饲料在改善猪肉品质中的应用。在本发明中，所述猪肉品质优选为育肥猪的猪肉品质。利用本发明所述L-苹果酸饲喂猪后能够在保育期显著增加猪骨骼肌的氧化型肌纤维比例。而且，本发明所述L-苹果酸对猪生长育肥期的生长性能无不良影响，对育肥猪的胴体性状无不良影响，提高屠宰后24h的pH值，降低滴水损失，可见，L-苹果酸对猪肉品质有很好的改善作用。
本发明提供了一种改善猪肉品质的方法，包括以下步骤：
在猪保育期饲喂上述技术方案所述的饲料。本发明在猪的保育期饲喂本发明所述饲料，能够影响保育期肌纤维类型转化进而改善猪肉品质。
肌纤维包括氧化型肌纤维和酵解型肌纤维，氧化型肌纤维直径细，含有丰富的线粒体和肌红蛋白，糖原含量低，有氧代谢能力强，增加氧化型肌纤维的比例会对肉色、pH值、嫩度、系水力等产生积极影响。本发明是通过影响窗口期(在猪出生后早期(出生至10周龄，即哺乳期和保育期))肌纤维类型转化进而改善猪肉品质，可见窗口期是影响肉品质的关键时期。
为了进一步说明本发明，下面结合附图和实施例对本发明提供的L-苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法进行详细地描述，但不能将它们理解为对本发明保护范围的限定。
实施例1
将2.5质量份(3750g)的L-苹果酸、3.75质量份(5625g)的蒙脱石和3.75质量份(5625g)的沸石粉混合后(L-苹果酸、蒙脱石和沸石粉的质量比为2.5：3.75:3.75)，得改善猪肉品质的混合物，即L-苹果酸混合物。
应用例1
选用96头28日龄三元杂交(杜×长×大)断奶仔猪，具有相似的遗传背景，随机分为两个处理，每个处理6个重复，每个重复8头猪(公母各半)。
设置两个处理，分别为：(1)对照组，饲喂基础日粮；(2)试验组，饲喂改善猪肉品质的饲料：以基础日粮的质量计，每kg基础日粮添加10g实施例1制备得到的L-苹果酸混合物；基础日粮以玉米-豆粕型日粮为基础，添加合成氨基酸，满足NRC(2012)推荐的猪体重为11～25kg阶段的营养需要。基础日粮的组分和营养水平见表1。
试验动物自由采食和饮水，试验周期42天。
表1猪保育期(11～25kg阶段)的基础日粮组成与营养水平(％，饲喂基础)
注：预混料向每千克饲料提供10000IU维生素A，1500IU维生素D3，30IU维生素E，2.5mg维生素K3，1.5mg维生素B1，10mg维生素B2，10mg维生素B6，0.05mg维生素B12，1mg叶酸，0.5mg生物素，30mg烟酸，20mg泛酸，20mg铜，100mg铁，110mg锌，40mg锰，0.3mg硒和0.54mg碘。
试验结束后，每个处理选取6头接近平均体重的猪屠宰，然后取背最长肌于-80℃保存，用于琥珀酸脱氢酶染色和实时荧光定量PCR。
其中，琥珀酸脱氢酶染色的具体方法如下：
①在37℃条件下使用琥珀酸脱氢酶孵育液(上海联硕生物科技有限公司)孵育冰冻切片(10μm)60min，去离子水漂洗2遍；
②随后使用4wt.％多聚甲醛固定15min，去离子水清洗2遍；
③2wt.％甲基绿染液复染2min，去离子水清洗2遍；
④酒精脱水，使用二甲苯透明化；
⑤使用中性树胶封片固定；
⑥使用倒置显微镜(CK40，Olympus公司，日本)观察切片。
琥珀酸脱氢酶染色结果如图1所示，与对照组相比，试验组背最长肌中琥珀酸脱氢酶阳性(深紫色)肌纤维的比例有升高的趋势(图1中的A和图1中的B，P＝0.07)，即图1A深紫色区域为琥珀酸阳性的肌纤维，即氧化型肌纤维。表明肌纤维类型向氧化型肌纤维转化。
实时荧光定量PCR的具体方法如下：
组织总RNA提取、反转录与cDNA合成、实时荧光定量PCR：参照Takara公司RNAisoPlus(9108)说明书提取背最长肌的RNA，参照Takara公司反转录试剂盒(RR047A)说明书进行反转录和cDNA合成。依据猪的mRNA序列，使用Primer 5.0软件设计引物并进行引物的合成，引物序列为表2所述的序列，内参基因是β-actin。cDNA经过10倍稀释之后用于实时荧光定量PCR。PCR反应体系见表3，反应条件见表4。采用2-ΔΔCt方法计算目的基因的相对表达水平。
表2实时荧光定量PCR引物序列
注：其中MyHC I为肌球蛋白重链Ⅰ，MyHC IIa为肌球蛋白重链Ⅱa，MyHC IIx为肌球蛋白重链Ⅱx，MyHC IIb为肌球蛋白重链Ⅱb，PGC-1α为过氧化物酶体增殖物激活受体γ共激活因子1，Myoglobin为肌红蛋白，Tnnt1为骨骼肌慢肌肌钙蛋白T，Tnnt3为骨骼肌慢肌肌钙蛋白T，β-actin为内参基因。
表3实时荧光定量PCR反应体系
| 组分 | 加样体积(μL) |
| 2×PCR反应试剂 | 5.00 |
| 上游引物 | 0.30 |
| 下游引物 | 0.30 |
| cDNA模板 | 0.40 |
| 灭菌超纯水 | 4.00 |
| 合计 | 10.00 |
表4实时荧光定量PCR反应条件
| 循环数 | 温度(℃) | 时间 | 说明 |
| 1 | 95 | 5min | 起始模板变性 |
| 95 | 15s | 模板变性 | |
| 35 | X | 15s | 退火 |
| 72 | 15s | 延伸并采集荧光信号 | |
| 1 | 60～95 | 6min | 溶解曲线 |
实时荧光定量PCR结果如表5所示。
表5 L-苹果酸混合物对保育期猪的肌纤维特异性基因表达水平的影响(n＝6)
由表5记载的可知，与对照组相比，试验组背最长肌中氧化型肌纤维标志性基因MyHC Ⅰ(肌球蛋白重链Ⅰ)、MyHC Ⅱa(肌球蛋白重链Ⅱa)、PGC-1α(过氧化物酶体增殖物激活受体γ共激活因子1)、Myoglobin(肌红蛋白)和Tnnt1(骨骼肌慢肌肌钙蛋白T)的mRNA表达水平显著升高(P<0.05)，中间型肌纤维标志性基因MyHCⅡx(肌球蛋白重链Ⅱx)的mRNA表达水平显著降低(P<0.05)，进一步说明了试验组骨骼肌的氧化型肌纤维比例显著提高。
应用例2
将应用例1中剩余84头猪按照保育期的分组，即对照组和试验组继续饲养。后续试验分为3个阶段，体重25～50kg为第1阶段，体重50～75kg为第2阶段，体重75～100kg为第3阶段。
3个阶段两个组全部饲喂基础日粮。基础日粮为玉米-豆粕型日粮，添加合成氨基酸，分别满足NRC(2012)推荐的生长育肥猪25～50kg、50～75kg和75～100kg的营养需要。基础日粮组成和营养水平见表6。
表6生长育肥期3个阶段的基础日粮组成与营养水平(％，饲喂基础)
试验动物自由采食和饮水，试验周期90天，计算平均日增重、平均日采食量和耗料增重比。试验结束后，每个处理选取6头接近平均体重的猪屠宰，检测胴体品质(胴体重、胴体长、屠宰率、眼肌面积、背膘厚、无脂瘦肉指数等)和肉品质(肉色、pH值、滴水损失、烹饪损失、剪切力等)，检测结果如表7～9所示。
表7保育期添加L-苹果酸混合物对生长育肥猪生长性能的影响(n＝6)
| 项目 | 对照组 | 试验组 | 标准误 | P值 |
| 平均日增重，g | 828 | 808 | 40.14 | 0.73 |
| 平均日采食量，g | 2382 | 2266 | 105.45 | 0.46 |
| 耗料增重比 | 2.92 | 2.81 | 0.17 | 0.67 |
表8保育期添加L-苹果酸混合物对育肥猪胴体性状的影响(n＝6)
表9保育期添加L-苹果酸混合物对育肥猪肉品质的影响(n＝6)
由表7～表9记载的可知，猪的保育期饲喂本发明所述饲料后，再采用应用例2提供的基础日粮饲喂，对育肥猪平均日增重、采食量、耗料增重比和胴体性状没有影响，且能够提高屠宰后24h的pH值，降低了滴水损失，降低了56.55％(P<0.05)。由此可见，在猪的保育期添加本发明所述含有L-苹果酸的饲料对猪生长育肥阶段的生长性能无不良影响，对育肥猪的胴体性状无不良影响，可以提高屠宰后24h的pH值，降低滴水损失，对猪肉品质有很好的改善作用。
由上述实施例记载可知，本发明所述L-苹果酸能够通过干预猪骨骼肌的肌纤维类型转化，达到提高猪骨骼肌氧化型肌纤维的比例的目的，提高屠宰后24h的pH值，降低滴水损失，进而达到改善肉品质的效果。同时对猪的安全性高，没有不良影响。
尽管上述实施例对本发明做出了详尽的描述，但它仅仅是本发明一部分实施例，而不是全部实施例，人们还可以根据本实施例在不经创造性前提下获得其他实施例，这些实施例都属于本发明保护范围。
序列表
<110> 中国农业大学
安徽雪郎生物科技股份有限公司
<120> L-苹果酸、混合物和饲料在改善猪肉品质中的应用和改善猪肉品质的方法
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Claims (9)
1.L-苹果酸在改善猪肉品质中的应用，其特征在于，所述L-苹果酸为唯一活性成分；
所述L-苹果酸结合基础日粮饲喂猪；
所述基础日粮由以下质量份的组分组成：玉米62.00~63.45份、豆粕9.65~11.00份、膨化全脂大豆9.10~10.45份、鱼粉3.80~4.22份、乳清粉5.76~6.21份、大豆浓缩蛋白1.90~2.10份、豆油1.72~1.76份、磷酸氢钙0.63~0.66份、石粉1.04~1.12份、食盐0.28~0.31份、L-赖氨酸盐酸盐0.58~0.61份、DL-蛋氨酸0.19~0.21份、L-苏氨酸0.22~0.25份、L-色氨酸0.04~0.07份、植酸酶0.01份和预混料0.50份；
所述改善猪肉品质包括提高猪骨骼肌氧化型肌纤维比例。
2.根据权利要求1所述的应用，其特征在于，所述猪肉品质为育肥猪的猪肉品质。
3.一种改善猪肉品质的饲料，其特征在于，所述饲料包括混合物和基础日粮；
所述混合物由L-苹果酸和载体组成；
所述基础日粮由以下质量份的组分组成：玉米62.00~63.45份、豆粕9.65~11.00份、膨化全脂大豆9.10~10.45份、鱼粉3.80~4.22份、乳清粉5.76~6.21份、大豆浓缩蛋白1.90~2.10份、豆油1.72~1.76份、磷酸氢钙0.63~0.66份、石粉1.04~1.12份、食盐0.28~0.31份、L-赖氨酸盐酸盐0.58~0.61份、DL-蛋氨酸0.19~0.21份、L-苏氨酸0.22~0.25份、L-色氨酸0.04~0.07份、植酸酶0.01份和预混料0.50份；
所述改善猪肉品质包括提高猪骨骼肌氧化型肌纤维比例。
4.根据权利要求3所述的饲料，其特征在于，所述载体包括蒙脱石和/或沸石粉。
5.根据权利要求3所述的饲料，其特征在于，所述L-苹果酸和载体的质量比为(2~3):(7~8)。
6.根据权利要求3所述的饲料，其特征在于，所述混合物与基础日粮的质量比为(0.5~1):(99~99.5)。
7.权利要求3~6任一项所述饲料在改善猪肉品质中的应用。
8.根据权利要求7所述的应用，其特征在于，所述猪肉品质为育肥猪的猪肉品质。
9.一种改善猪肉品质的方法，其特征在于，包括以下步骤：
在猪保育期饲喂权利要求3~6任一项所述的饲料。
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