Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 8: 5067 - 5075

DOI: https://doi.org/10.12418/CJAN2024.431

RESEARCH PAPER

Effects of Vitamin B12 on Laying Performance, Egg Quality, Serum Biochemical and Antioxidant Indices of Wulong Geese during Laying Period

  • TAN Shaoxing , 1 ,
  • WANG Baowei , 2, * ,
  • ZHANG Ming’ai 1 ,
  • FAN Wenlei 1 ,
  • YUE Bin 2 ,
  • KONG Min 2 ,
  • WANG Binghan 3
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  • 1 College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
  • 2 Institute of High Quality Waterfowl, Qingdao Agricultural University, Qingdao 266109, China
  • 3 Qingdao Huihe Biotechnology Co., Ltd., Qingdao 266109, China
*professor, E-mail:

Received date: 2024-02-16

  Online published: 2024-08-12

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Abstract

The purpose of this study was to study the effects of dietary different supplemental levels of vitamin B12 on laying performance, egg quality, serum biochemical and antioxidant indices of Wulong geese during laying period. A total of 180 thirty-four-week-old Wulong geese were randomly divided into 6 groups with 6 replicates in each group and 5 geese in each replicate (1 male and 4 females). The vitamin B12 supplemental levels in each group were 0 (control group), 12.5, 25.0, 37.5, 50.0 and 62.5 μg/kg, respectively. The pre-experimental period lasted for 1 week, and the experimental period lasted for 16 weeks. The results showed as follows: 1) compared with the control group, the average daily feed intake of 12.5, 25.0, 37.5, 50.0 and 62.5 μg/kg vitamin B12 group was significantly increased (P<0.05), and the laying rate and egg number of 25.0, 37.5 and 62.5 μg/kg vitamin B12 group were significantly increased (P<0.05). 2) Compared with the control group, the eggshell strength of 12.5, 25.0, 37.5, 50.0 and 62.5 μg/kg vitamin B12 group was significantly increased (P<0.05), and the eggshell thickness of 25.0, 37.5, 50.0 and 62.5 μg/kg vitamin B12 group was significantly increased (P<0.05). 3) Compared with the control group, the contents of total cholesterol (T-CHO) and low density lipoprotein cholesterol (LDL-C) in serum of 12.5, 25.0, 37.5, 50.0 and 62.5 μg/kg vitamin B12 group were significantly decreased (P<0.05), and the serum albumin (ALB) content of 25.0 and 37.5 μg/kg vitamin B12 group was significantly increased (P<0.05). 4) Compared with the control group, the serum total antioxidant capacity (T-AOC) of 25.0 and 37.5 μg/kg vitamin B12 group was significantly increased (P<0.05), and the serum glutathione peroxidase (GSH-Px) activity of 12.5, 25.0, 37.5, 50.0 and 62.5 μg/kg vitamin B12 group was significantly increased (P<0.05). In conclusion, dietary vitamin B12 can improve the laying performance and egg quality of Wulong geese during laying period, regulate the serum biochemical indices, and enhance the antioxidant capacity. Under the conditions of this experiment, the dietary vitamin B12 suitable supplemental level is 25.0 μg/kg.

Key words： vitamin B12; Wulong geese during laying period; laying performance; egg quality; serum biochemical indices; antioxidant capacity

Cite this article

TAN Shaoxing , WANG Baowei , ZHANG Ming’ai , FAN Wenlei , YUE Bin , KONG Min , WANG Binghan . Effects of Vitamin B12 on Laying Performance, Egg Quality, Serum Biochemical and Antioxidant Indices of Wulong Geese during Laying Period[J]. Chinese Journal of Animal Nutrition, 2024 , 36(8) : 5067 -5075 . DOI: 10.12418/CJAN2024.431

五龙鹅是一种小型高产鹅品种,具有耐粗饲、抗病力强等特点,母鹅一般在8~9月龄开始产蛋,年产蛋120个左右,平均蛋重约128.5 g[1],具有很大的开发潜力。维生素B12(vitamin B12)即钴胺素,是动物体内唯一含有金属元素的一种维生素,它在人体一碳基团的代谢中起着转移甲基的作用[2]。维生素B12在调控动物神经发育、生殖、养分代谢等方面发挥重要作用,饲粮中添加适宜水平维生素B12对畜禽生长发育、繁殖后代、机体健康方面都有积极作用[3]。此外,维生素B12通过清除自由基和减少氧化应激来保护DNA免受活性氧造成的损伤,有一定的抗氧化作用[4]。研究表明,维生素B12缺乏不仅会影响家禽代谢活动,而且会造成蛋重降低、产蛋率下降,长时间缺乏还会导致食欲不振及减少采食量[5]。维生素B12缺乏也会影响家畜的配种,从而无法正常繁殖后代[6];同时,对家畜妊娠也会有不利影响,可能出现流产、仔猪体弱多病甚至死亡[7]。龙建华等[8]研究表明,在5~15周龄五龙鹅饲粮中添加不同水平维生素B12对五龙鹅的终末体重和平均日增重有显著影响,且在添加水平为0.01 mg/kg时较高。王瑞[9]研究表明,京红蛋鸡饲粮中添加50、100、400 μg/kg的维生素B12可以显著提高蛋壳厚度(eggshell thickness,ET),调节血清脂质代谢,提高血清白蛋白(albumin,ALB)含量。刘欢等[10]研究表明,与对照组相比,饲粮中添加维生素B12组的灶性脑缺血大鼠的血清超氧化物歧化酶(superoxide dismutase,SOD)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性显著上升,血清丙二醛(malondialdehyde,MDA)含量显著降低。目前,维生素B12在家禽中的研究主要以鸡和鸭为主,而关于产蛋期五龙鹅饲粮中适宜维生素B12添加水平缺乏数据参数。因此,本试验选用产蛋期五龙鹅为试验对象,在产蛋期五龙鹅饲粮中添加不同水平的维生素B12,以产蛋期五龙鹅的产蛋性能、蛋品质、血清生化和抗氧化指标为评价指标,筛选产蛋期五龙鹅饲粮中维生素B12的适宜添加水平,为进一步完善产蛋期五龙鹅的营养需求数据提供参考。

1 材料与方法

1.1 试验设计

选择34周龄产蛋期五龙鹅180只,随机分为6组,每组6个重复,每个重复5只鹅(1公4母)。各组饲粮中维生素B12添加水平分别为0(对照组)、12.5、25.0、37.5、50.0、62.5 μg/kg。预试期1周,正试期16周。试验用维生素B12有效含量为1%。

1.2 试验饲粮

采用玉米-豆粕型基础饲粮,参考我国种鹅生产实际设计。基础饲粮组成及营养水平见表1。基础饲粮中粗纤维含量参照GB/T 6434—2006[11]测定,粗蛋白质含量参照GB/T 6432—2018[12]测定,维生素B12含量参照GB/T 17819—2017[13]测定。
表1 基础饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of the basal diet (air-dry basis) %

项目Items 含量Content
原料Ingredients
玉米Corn 56.20
豆粕Soybean meal 23.40
麸皮Bran 4.00
菊花梗粉Chrysanthemum stalk powder 4.80
豆油Soybean oil 3.00
石粉Limestone 6.00
磷酸氢钙CaHPO4 1.50
食盐NaCl 0.40
微量元素预混料Trace elements premix1) 0.50
维生素预混料Vitamin premix1) 0.10
蛋氨酸Methionine 0.10
合计Total 100.00
营养水平Nutrient levels2)
代谢能ME/(MJ/kg) 11.43
粗蛋白质CP 16.41
粗纤维CF 4.54
钙Ca 2.71
总磷TP 0.60
蛋氨酸Met 0.35
赖氨酸Lys 0.89
苏氨酸Thr 0.61
色氨酸Trp 0.22
维生素B12 VB12/(μg/kg) 0.00

1)微量元素预混料和维生素预混料为每千克饲粮提供 Trace elements premix and vitamin premix provided the following per kg of the diet:Mn 30 mg,Zn 65 mg,I 0.3 mg,Fe 80 mg,Se 0.5 mg,Cu 4 mg,叶酸 folic acid 0.5 mg,VD3 2 000 IU,生物素 biotin 0.2 mg,VK3 0.8 mg,VA 9 000 IU,VE 40 mg,VB1 2.0 mg,烟酰胺 nicotinamide 30 mg,VB2 4.0 mg,胆碱 choline 740 mg/kg,泛酸钙 calcium pantothenate 11 mg,VB6 4.0 mg。

2)维生素B12、粗蛋白质和粗纤维为实测值,其余均为计算值。VB12, CP and CF were measured values, while the others were calculated values.

1.3 饲养管理

每个重复提供舒适的产蛋箱,鹅舍为半开放式舍,采用地面平养的方法饲养试验鹅,光照为自然光照。试验过程保证试验鹅自由采食和自由饮水,每周清扫鹅舍2次并且严格消毒。

1.4 采集样品与指标测定

1.4.1 产蛋性能和蛋品质指标

试验期间,每天记录各重复饲料消耗量,并收集各重复的鹅蛋,记录数量和称量蛋重。试验末期,每个重复收集4枚种蛋,每组24枚,共144枚,待测蛋品质。参考《家禽生产性能名词术语和度量统计方法》(NY/T 823—2020)[14]测定产蛋性能和蛋品质指标。通过游标卡尺测量蛋形指数(egg shape index,ESI),通过ETG-1061蛋壳厚度测量仪测量蛋壳厚度,通过EFG-0503蛋壳强度测定仪测定蛋壳强度(eggshell strength,ES),通过EMT-7300蛋品质测定仪测定蛋白高度(albumen height,AH)、蛋黄颜色(yolk color,YC)、哈氏单位(Haugh unit,HU)和蛋黄比率(yolk ratio,YR)。

1.4.2 血清生化和抗氧化指标

试验结束时,随机选36只母鹅(每个重复1只),采翅静脉血10 mL,离心机预热0.5 h, 4 ℃、3 000 r/min离心30 min,取上清分装并标号,将分装好的血清样本置于-40 ℃冰箱中保存。血清甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,T-CHO)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)和高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)含量采用比色法测定,ALB含量采用溴甲酚绿法测定,总蛋白(total protein,TP)含量采用考马斯亮蓝法测定,过氧化氢酶(catalase,CAT)活性采用钼酸铵法测定;总超氧化物歧化酶(total superoxide dismutase,T-SOD)活性采用羟胺法测定;总抗氧化能力(total antioxidant capacity,T-AOC)采用2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)法测定,MDA含量采用硫代巴比妥酸(TBA)法测定,GSH-Px活性采用比色法测定。

1.5 统计分析

试验数据采用SPSS 25.0软件进行单因素方差分析(one-way ANOVA),并采用Duncan氏法进行组间多重比较。结果用平均值和均值标准误(SEM)表示,P<0.05为差异显著。

2 结果

2.1 维生素B12对产蛋期五龙鹅产蛋性能的影响

由表2的结果可以看出,与对照组相比,12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组的平均日采食量显著升高(P<0.05),25.0、37.5、62.5 μg/kg维生素B12组的产蛋率和产蛋数显著升高(P<0.05),并且25.0 μg/kg维生素B12组的产蛋率和产蛋数最高。各组之间平均蛋重和料蛋比均无显著差异(P>0.05)。
表2 维生素B12对产蛋期五龙鹅产蛋性能的影响

Table 2 Effects of vitamin B12 on laying performance of Wulong geese during laying period

项目
Items		 维生素B12添加水平Vitamin B12 supplemental levels/(μg/kg) SEM P值
P-value
0(对照
Control)		 12.5 25.0 37.5 50.0 62.5
产蛋率LR/% 22.47c 25.07bc 33.11a 31.18ab 28.27abc 31.32ab 1.18 0.035
平均日采食量ADFI/(g/d) 166.98b 187.38a 181.35a 186.26a 183.61a 185.77a 9.02 0.022
平均蛋重AEW/g 115.67 122.37 118.95 126.31 122.34 122.42 5.99 0.388
料蛋比F/E 6.69 6.14 4.63 4.84 5.44 4.88 1.15 0.151
产蛋数EN/(个/只) 25.17c 28.08bc 37.08a 34.92ab 31.67abc 35.08ab 1.32 0.035
产蛋总重EM/(kg/只) 2.93 3.44 4.42 4.43 3.90 4.30 0.81 0.098

同行数据肩标无字母或相同字母表示差异不显著(P>0.05),不同小写字母表示差异显著(P<0.05)。下表同。

In the same row, values with no letter or the same letter superscripts mean no significant difference (P>0.05), while with different small letter superscripts mean significant difference (P<0.05). The same as below.

2.2 维生素B12对产蛋期五龙鹅蛋品质的影响

由表3的结果可以看出,与对照组相比,12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组的蛋壳强度显著升高(P<0.05),25.0、37.5、50.0、62.5 μg/kg维生素B12组的蛋壳厚度显著升高(P<0.05),并且25.0 μg/kg维生素B12组的蛋壳强度和蛋壳厚度最高。各组之间蛋形指数、蛋白高度、哈氏单位、蛋黄颜色和蛋黄比率均无显著差异(P>0.05)。
表3 维生素B12对产蛋期五龙鹅蛋品质的影响

Table 3 Effects of vitamin B12 on egg quality of Wulong geese during laying period

项目
Items		 维生素B12添加水平Vitamin B12 supplemental levels/(μg/kg) SEM P值
P-value
0(对照
Control)		 12.5 25.0 37.5 50.0 62.5
蛋形指数ESI 1.48 1.48 1.43 1.46 1.43 1.47 0.01 0.153
蛋壳强度ES/(kg/cm2) 4.80b 5.02a 5.12a 5.08a 5.06a 5.03a 0.03 0.030
蛋壳厚度ET/mm 0.50b 0.53ab 0.56a 0.55a 0.54a 0.54a 0.01 0.030
蛋白高度AH/mm 14.54 15.07 15.75 15.31 15.33 15.48 0.30 0.912
哈氏单位HU 107.52 109.38 111.36 109.97 109.31 110.26 1.16 0.966
蛋黄颜色YC 4.34 4.44 4.58 4.49 4.97 4.64 0.15 0.895
蛋黄比率YR/% 29.36 29.49 29.81 30.46 31.55 30.21 0.38 0.588

2.3 维生素B12对产蛋期五龙鹅血清生化指标的影响

由表4的结果可以看出,与对照组相比,12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组的血清T-CHO和LDL-C含量显著降低(P<0.05),25.0和37.5 μg/kg维生素B12组的血清ALB含量显著升高(P<0.05)。各组之间血清TG、HDL-C和TP含量均无显著差异(P>0.05)。
表4 维生素B12对产蛋期五龙鹅血清生化指标的影响

Table 4 Effects of vitamin B12 on serum biochemical indices of Wulong geese during laying period

项目
Items		 维生素B12添加水平Vitamin B12 supplemental levels/(μg/kg) SEM P值
P-value
0(对照
Control)		 12.5 25.0 37.5 50.0 62.5
甘油三酯TG/(mmol/L) 1.10 0.72 0.69 0.78 0.69 0.72 0.09 0.821
总胆固醇T-CHO/(mmol/L) 5.59a 3.68b 3.45b 3.50b 3.35b 3.12b 0.24 0.016
低密度脂蛋白胆固醇
LDL-C/(mmol/L)		 1.06a 0.68b 0.65b 0.55b 0.49b 0.46b 0.06 0.031
高密度脂蛋白胆固醇
HDL-C/(mmol/L)		 1.26 1.43 1.63 1.51 1.56 1.53 0.05 0.472
白蛋白ALB/(g/L) 12.14c 12.28c 16.33a 15.87ab 14.95abc 12.57bc 0.53 0.033
总蛋白TP/(g/L) 40.83 42.57 51.80 48.96 47.98 44.53 1.67 0.419

2.4 维生素B12对产蛋期五龙鹅血清抗氧化指标的影响

由表5的结果可以看出,与对照组相比,25.0和37.5 μg/kg维生素B12组的血清T-AOC显著升高(P<0.05),12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组的血清GSH-Px活性显著升高(P<0.05),并且25.0 μg/kg维生素B12组的血清T-AOC和GSH-Px活性最高。各组之间血清CAT、T-SOD活性和MDA含量均无显著差异(P>0.05)。
表5 维生素B12对产蛋期五龙鹅血清抗氧化指标的影响

Table 5 Effects of vitamin B12 on serum antioxidant indices of Wulong geese during laying period

项目
Items		 维生素B12添加水平Vitamin B12 supplemental levels/(μg/kg) SEM P值
P-value
0(对照
Control)		 12.5 25.0 37.5 50.0 62.5
丙二醛MDA/(nmol/mL) 3.61 3.47 2.07 2.42 2.66 2.77 1.14 0.445
过氧化氢酶CAT/(U/mL) 15.42 15.56 17.24 16.90 16.79 16.33 1.07 0.056
总抗氧化能力
T-AOC/(mmol/L)		 0.24c 0.40bc 0.86a 0.65ab 0.50abc 0.47bc 0.29 0.031
总超氧化物歧化酶
T-SOD/(U/mL)		 153.75 176.36 264.54 248.21 239.18 210.17 15.72 0.281
谷胱甘肽过氧化物酶
GSH-Px/(U/mL)		 201.98b 259.85a 308.04a 296.97a 280.20a 268.97a 10.01 0.018

3 讨论

3.1 维生素B12对产蛋期五龙鹅产蛋性能的影响

生产性能是畜禽生产的重要指标,维生素B12对其有重要的作用。侯水生等[15]研究表明,与对照组相比,饲粮中添加不同水平维生素B12组肉鸡增重和采食量显著提高,料重比显著降低,且维生素B12添加水平为0.024 mg/kg时效果最好。Keating[16]研究表明,水蚤的繁殖率随培养基中维生素B12的浓度而增加,当培养基中维生素B12浓度为0.75~1.00 μg/kg时,水蚤后代数量最多。El-Katcha等[17]研究表明,在蛋鸡饲粮中添加20 μg/kg维生素B12可以显著提高平均产蛋量。Keshavarz[18]研究表明,蛋鸡产蛋期后期在减少饲粮中蛋白质水平的情况下,不添加叶酸和维生素B12,会降低蛋鸡的蛋重和产蛋量。Squires等[19]研究发现,随着饲粮中维生素B12添加水平的增加,母鸡的产蛋量和蛋重提高,当饲粮中维生素B12添加水平为8 μg/kg时,可获得最大产蛋量和蛋重。本试验研究发现,与对照组相比,12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组五龙鹅的平均日采食量显著升高,25.0、37.5、62.5 μg/kg维生素B12组的产蛋率和产蛋数显著升高,其中25 μg/kg维生素B12组的产蛋率和产蛋数最高。这可能是因为维生素B12与叶酸一起参与体内蛋氨酸循环,从而影响甲基的转移,在维生素B12的参与下,蛋氨酸合成酶使叶酸将一碳单位转移给高半胱氨酸,使其得到甲基而转变为蛋氨酸[20],蛋氨酸含量增加,从而使五龙鹅的产蛋量增加。此外,维生素B12可以使优势卵泡更快生长,为排卵做更好的准备[21],且补充维生素B12可以通过减少线粒体应激来增强蛋氨酸介导的卵黄生成和卵母细胞发育[22],从而提高产蛋期五龙鹅的产蛋性能。

3.2 维生素B12对产蛋期五龙鹅蛋品质的影响

蛋品质是家禽产品的重要指标,包括蛋的外形与内容物的品质,受遗传、饲养管理、蛋贮存期等因素影响。Janist等[23]研究表明,在蛋鸡饲粮中添加叶酸和维生素B12有提高鸡蛋蛋壳强度和蛋壳厚度的趋势。武笑天[24]研究表明,京红蛋鸡饲粮中添加不同水平的维生素B12,其蛋壳厚度和哈氏单位有明显差异,饲粮中维生素B12添加水平为25 μg/kg时,蛋壳厚度和哈氏单位均达到最高值。何欣等[25]研究表明,饲粮中添加维生素B12对改善33周龄蛋鸡的蛋壳品质有促进作用。本试验研究发现,与对照组相比,12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组的蛋壳强度显著升高,25.0、37.5、50.0、62.5 μg/kg维生素B12组的蛋壳厚度显著升高,各组之间蛋白高度和哈氏单位虽然没有显著差异,但添加维生素B12组的蛋白高度和哈氏单位均有所提升,并且以上及指标均在饲粮中维生素B12添加水平为25.0 μg/kg时最优,与前人结果基本一致。由此可见,在产蛋期五龙鹅饲粮中添加维生素B12有利于提高蛋品质,具体机制有待进一步探究。

3.3 维生素B12对产蛋期五龙鹅血清生化指标的影响

脂质在细胞中起着维持体内平衡的重要作用,血液脂质产生ATP并参与合成维生素等多种生理反应[26],ALB、TP是反映机体营养状况和炎症状态的重要指标,对机体应激状态有重要作用[27]。Boachie等[28]研究表明,与正常组相比,人肝癌细胞(HepG2)在维生素B12缺乏组培养基中脂肪酸合成增加,氧化减少,脂质代谢失调。张大龙等[29]研究发现,维生素B12能有效促进高脂饮食大鼠的血脂代谢。李庆云等[30]研究表明,1~42日龄北京鸭的夏季饲粮中添加高水平维生素B12有提高其血清ALB含量的趋势。Ahmad[31]研究发现,饲粮中添加不同水平的维生素B12对1~21日龄北京鸭血清TP和ALB含量无显著影响。本试验研究发现,与对照组相比,12.5、25.0、37.5、50.0和62.5 μg/kg维生素B12组的血清T-CHO和LDL-C含量显著降低,25.0和37.5 μg/kg维生素B12组的血清ALB含量显著升高,推测可能是因为维生素B12在甲基丙二酰辅酶A转化为琥珀酰辅酶A的过程中充当辅酶,如果维生素B12含量低,该反应就会被阻断,导致甲基丙二酰辅酶A积累,从而抑制脂肪酸氧化的限速酶,引起脂肪含量升高,进而影响血脂水平[32]。另外,植物性蛋白质在维生素B12的参与下利用率得到提高,而且维生素B12可以促进蛋白质和脂肪代谢等,并且参与氨基酸和甲基嘌呤的合成[33],进而调节产蛋期五龙鹅血清生化指标。

3.4 维生素B12对产蛋期五龙鹅血清抗氧化指标的影响

MDA目前被认为是氧化脂质损伤使用最广泛的代表[34]。GSH-Px、SOD和CAT是公认的抗氧化酶标志物,可以减轻活性氧负荷并作为抗氧化防御系统[35]。曲木等[36]研究发现,维生素B12能显著提高黄颡鱼的抗氧化能力,在饲料中加入0.15%的维生素B12时,其血清CAT、T-SOD活性最高,而MDA含量最低。Misra等[37]研究发现,与对照组相比,维生素B12缺乏症患者的血清GSH-Px活性和T-AOC降低,MDA含量升高。本试验研究发现,随着饲粮中维生素B12添加水平增加,产蛋期五龙鹅血清T-AOC呈先升高后降低趋势;12.5、25.0、37.5、50.0、62.5 μg/kg组的血清GSH-Px活性显著升高,且在维生素B12添加水平为25.0 μg/kg时效果最佳,与上述结果基本一致。这可能与维生素B12可以直接清除活性氧,特别是超氧化物,通过保存谷胱甘肽间接刺激活性氧清除,调节细胞因子和生长因子的产生,以提供对免疫反应诱导的氧化应激的保护,降低同型半胱氨酸诱导的氧化应激,减少晚期糖基化终产物引起的氧化应激有关[38],从而改善产蛋期五龙鹅血清抗氧化指标。

4 结论

饲粮中添加维生素B12可提高产蛋期五龙鹅的产蛋性能、蛋品质,调节血清生化指标,增强抗氧化能力。本试验条件下,饲粮中维生素B12适宜添加水平为25.0 μg/kg。

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