研究论文

饲粮精氨酸水平对产蛋期五龙鹅种蛋质量、血清激素和细胞因子含量的影响

  • 陈莹 ,
  • 张贝贝 ,
  • 王宝维 ,
  • 张名爱 ,
  • 凡文磊 ,
  • 李文立 , *
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  • 青岛农业大学动物科技学院,青岛 266109
*李文立,教授,博士生导师,E-mail:

陈莹(1998—),女,山东烟台人,硕士研究生,研究方向为动物营养与饲料科学。E-mail:

Office editor: 武海龙

收稿日期: 2023-04-21

  网络出版日期: 2023-10-12

基金资助

国家现代农业产业技术体系(CARS-42-14)

荣昌市农牧业重点研发项目(cstc2020ngzx0007)

山东省重点技术研发计划项目(2021CXGC010805)

山东省自然科学基金青年基金(ZR2020QC181)

Effects of Dietary Arginine Level on Breeding Egg Quality, Serum Hormone and Cytokine Contents of Wulong Geese during Laying Period

  • CHEN Ying ,
  • ZHANG Beibei ,
  • WANG Baowei ,
  • ZHANG Ming’ai ,
  • FAN Wenlei ,
  • LI Wenli , *
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  • College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
*professor, E-mail:

Received date: 2023-04-21

  Online published: 2023-10-12

摘要

本试验旨在研究饲粮精氨酸水平对产蛋期五龙鹅种蛋质量、血清激素和细胞因子含量的影响。试验选取34周龄体况相近的产蛋期五龙鹅150只,随机分成6个组,每组5个重复,每个重复5只(1公4母)。各组分别饲喂精氨酸水平为0.72%(对照组)、0.82%、0.92%、1.02%、1.12%和1.22%的试验饲粮。预试期1周,正试期16周。结果表明:1)0.82%、0.92%、1.02%、1.12%、1.22%组种蛋孵化率(HR)显著高于对照组(P<0.05),0.92%、1.02%组种蛋哈氏单位(HU)显著高于对照组(P<0.05),1.02%组种蛋蛋壳厚度(ET)显著高于对照组(P<0.05)。2)1.02%组血清胰岛素(INS)和孕酮(P4)含量显著高于对照组(P<0.05)。3)0.92%、1.02%组血清白细胞介素-1 (IL-1 )含量显著低于对照组(P<0.05),0.92%、1.02%组血清白细胞介素-4(IL-4)含量显著高于对照组(P<0.05)。由此可见,饲粮适宜精氨酸水平可提高产蛋期五龙鹅种蛋质量,调节激素水平,增强机体免疫功能。在本试验条件下,饲粮适宜精氨酸水平为1.02%。

本文引用格式

陈莹 , 张贝贝 , 王宝维 , 张名爱 , 凡文磊 , 李文立 . 饲粮精氨酸水平对产蛋期五龙鹅种蛋质量、血清激素和细胞因子含量的影响[J]. 动物营养学报, 2023 , 35(10) : 6414 -6422 . DOI: 10.12418/CJAN2023.588

Abstract

This experiment was conducted to investigate the effects of dietary arginine level on breeding egg quality, serum hormone and cytokine contents of Wulong geese during laying period. A total of 150 thirty-four-week-old Wulong geese during laying period with similar body condition were randomly divided into 6 groups with 5 replicates per group and 5 geese per replicate (1 male and 4 female). Geese in 6 groups were fed experimental diets which the arginine levels were 0.72% (control group), 0.82%, 0.92%, 1.02%, 1.12% and 1.22%, respectively. The pre-trial period lasted for 1 week, and the experimental period lasted for 16 weeks. The results showed as follows: 1) the breeding egg hatching rate of 0.82%, 0.92%, 1.02%, 1.12% and 1.22% groups was significantly higher than that of the control group (P<0.05), the breeding egg Haugh unit of 0.92% and 1.02% groups was significantly higher than that of the control group (P<0.05), and the breeding egg eggshell thickness of 1.02% group was significantly higher than that of the control group (P<0.05). 2) The contents of insulin (INS) and progesterone (P4) in serum of 1.02% group were significantly higher than those of the control group (P<0.05). 3) The serum interleukin-1β (IL-1β) content of 0.92% and 1.02% group was significantly lower than that of the control group (P<0.05), and the serum interleukin-4 (IL-4) content of 0.92% and 1.02% group was significantly higher than that of the control group (P<0.05). In conclusion, dietary optimal arginine level can improve the breeding egg quality, regulate hormone level, and enhance the body immune function of Wulong geese during laying period. Under the conditions of this experiment, the dietary optimal arginine level is 1.02%.[Chinese Journal of Animal Nutrition, 2023, 35(10):6414-6422]

五龙鹅又称豁眼鹅,具有体型小、产蛋量高、抗病力强、耐粗饲等优点[1],开发利用潜力大。目前,国内外有关种鹅营养需要参数的研究较少,尚未颁布种鹅的饲养标准,导致实际生产中饲料配制缺乏依据,在一定程度上影响了种鹅的产蛋性能和繁殖性能。
精氨酸(arginine,Arg)是一种碱性氨基酸,在自然界中以D型和L型2种形式存在,畜禽体内主要以L-精氨酸为主[2]L-精氨酸是一种多用途氨基酸[3],是多种生物学效应化合物一氧化氮(NO)、脯氨酸、多胺、谷氨酸等的前体,在畜禽营养转运蛋白表达、免疫调节、繁殖生产等方面发挥重要作用[4-6]。在家禽代谢途径中,由于精氨酸合成所需的关键酶活性有限[7],不能有效自主合成精氨酸,只能从饲粮中额外获取满足以自身需求,因此,精氨酸常被称为家禽的第四限制性氨基酸[8]L-精氨酸作为蛋白质链中的单体合成蛋白质,参与多种代谢途径[9],并参与畜禽生长发育、繁殖以及免疫等过程。研究表明,肉鸡饲粮中添加0.96%~1.27%的精氨酸可提高种蛋受精率(fertilization rate,FR)、孵化率(hatching rate,HR)[10]和哈氏单位(Haugh unit,HU)[2]。饲粮中添加1.0%的精氨酸显著降低了低出生重仔猪肝脏白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)mRNA表达量,作为免疫调节介质调控免疫,增强了仔猪免疫力[11]。目前,国内外有关鹅精氨酸需要量的研究很少,家禽主要集中鸡和鸭,而关于鹅饲粮精氨酸水平未有确切标准,所以鹅饲粮适宜精氨酸水平的试验研究对填补五龙鹅精氨酸营养需要的空白具有重要意义。本课题组前期研究也发现,饲粮中1.02%的精氨酸水平可促进产蛋期五龙鹅的产蛋率和平均蛋重[12]。因此,本试验以产蛋期五龙鹅为试验对象,研究饲粮精氨酸水平对五龙鹅种蛋质量、血清激素和细胞因子含量的影响,探究产蛋期五龙鹅饲粮适宜精氨酸水平,为精氨酸在产蛋期种鹅生产应用中提供理论依据。

1 材料与方法

1.1 试验饲粮

试验用精氨酸为L-精氨酸(有效含量为99%),购自青岛某生物科技有限公司。饲粮配方参考我国鹅生产实际和NRC(1994)家禽营养标准设计,原料以玉米和玉米蛋白粉为主。试验饲粮精氨酸含量参考GB/T 18246—2019测定。试验饲粮组成及营养水平见表1
表1 试验饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of experimental diets (air-dry basis)%

项目
Items
饲粮精氨酸水平 Dietary arginine level/%
0.72 0.82 0.92 1.02 1.12 1.22
原料 Ingredients
玉米 Corn 64.12 64.12 64.12 64.12 64.12 64.12
玉米蛋白粉 Corn gluten meal 11.10 11.10 11.10 11.10 11.10 11.10
菜籽粕 Rapeseed meal 5.10 5.10 5.10 5.10 5.10 5.10
麸皮 Bran 3.99 3.99 3.99 3.99 3.99 3.99
菊花梗粉 Chrysanthemum stalks powder 2.89 2.89 2.89 2.89 2.89 2.89
豆油 Soybean oil 2.20 2.20 2.20 2.20 2.20 2.20
豆粕 Soyabean meal 1.95 1.95 1.95 1.95 1.95 1.95
石粉 Limestone 5.65 5.65 5.65 5.65 5.65 5.65
磷酸氢钙 CaHPO4 1.28 1.28 1.28 1.28 1.28 1.28
食盐 NaCl 0.34 0.34 0.34 0.34 0.34 0.34
蛋氨酸 Met 0.11 0.11 0.11 0.11 0.11 0.11
预混料 Premix1) 0.77 0.77 0.77 0.77 0.77 0.77
L-丙氨酸 L-Ala 0.50 0.40 0.30 0.20 0.10 0.00
L-精氨酸 L-Arg 0.10 0.20 0.30 0.40 0.50
合计 Total 100.00 100.00 100.00 100.00 100.00 100.00
营养水平 Nutrient levels2)
代谢能 ME/(MJ/kg) 12.01 12.01 12.01 12.01 12.01 12.01
粗蛋白质 CP 18.03 18.03 18.03 18.03 18.03 18.03
钙 Ca 2.27 2.27 2.27 2.27 2.27 2.27
总磷 TP 0.66 0.66 0.66 0.66 0.66 0.66
L-精氨酸 L-Arg 0.72 0.82 0.92 1.02 1.12 1.22
赖氨酸 Lys 0.73 0.73 0.73 0.73 0.73 0.73
蛋氨酸 Met 0.47 0.47 0.47 0.47 0.47 0.47
色氨酸 Trp 0.15 0.15 0.15 0.15 0.15 0.15
苏氨酸 Thr 0.57 0.57 0.57 0.57 0.57 0.57

1)预混料为每千克饲粮提供 Premix provided the following per kg of diets:VA 9 000 IU,VD3 2 000 IU,VE 40 mg,VB1 2.0 mg,VB2 4.0 mg,VB6 4.0 mg,VB11 0.5 mg,VB12 12 μg,VK3 0.8 mg,Zn 65 mg,Cu 4.0 mg,I 0.3 mg,Mn 30 mg,烟酰胺 nicotinic acid 30 mg,赖氨酸 Lys 2.5 g,Fe 80 mg,Se 0.5 mg,泛酸钙 calcium pantothenate 11 mg,生物素 biotin 0.2 mg,胆碱 choline 740 mg。

2)L-精氨酸、粗蛋白质、钙和总磷为实测值,其余营养水平为计算值。L-Arg, CP, Ca and TP were measured values, while the others were calculated values.

1.2 试验设计

选取34周龄体况相近的产蛋期五龙鹅150只,随机分成6个组,每组5个重复,每个重复5只(1公4母)。各组分别饲喂精氨酸水平为0.72%(对照组)、0.82%、0.92%、1.02%、1.12%和1.22%的试验饲粮。试验从2022年3月7日开始,预试期1周,正试期16周。

1.3 饲养管理

试验开始前对鹅舍进行充分冲洗和严格消毒,保证鹅舍水槽及料槽干净卫生。每重复为1栏,栏与栏之间通过铁丝网进行隔离。正试期内,每周进行2次杀菌消毒。整个试验期,试验鹅自由饮水、自由采食,地面开放式饲养。每日注意观察鹅群生长状况。

1.4 测定指标与方法

1.4.1 种蛋质量

试验8周后,每重复收集10枚种蛋进行孵化,共300枚,记录受精蛋数、出雏数等,计算HR、FR和合格率(qualified rate,QR),计算公式如下:
HR(%)=(出雏数/受精种蛋数)×100;
FR(%)=(合格种蛋数/产蛋总数)×100;
QR(%)=(受精种蛋数/入孵种蛋数)×100。
试验第16周,每组收集25枚(每重复5枚),共150枚种蛋,用于测定蛋品质指标,蛋形指数(egg shape index,ESI)通过游标卡尺量取;蛋壳厚度(eggshell thickness,ET)使用厚度测定仪测量;蛋黄颜色(yolk color,YC)、蛋白高度(albumen height,AH)、HU、蛋黄比率(yolk ratio,YR)采用蛋品质分析仪(DET-6000,NABEL Co., Ltd.,日本)测定。

1.4.2 血清激素和细胞因子含量

试验期结束后,每重复随机选择1只母鹅,翅根静脉采血10 mL,储存在促凝剂采血管中。4 ℃条件下,3 000 r/min离心15 min分离上清液,-80 ℃冰箱保存。采用双抗体一步夹心法酶联免疫吸附试验测定血清中胰岛素(INS)、促黄体生成素(LH)、促卵泡素(FSH)、孕酮(P4)以及IL-1β、白细胞介素-2(IL-2)、IL-6、白细胞介素-4(IL-4)、白细胞介素-10(IL-10)含量。以上试剂盒均购自南京建成生物工程研究所。

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氏法进行多重比较。试验结果用“平均值±标准误”表示,P<0.05表示差异显著。

2 结果

2.1 饲粮精氨酸水平对产蛋期五龙鹅种蛋质量的影响

表2可知,0.82%、0.92%、1.02%、1.12%、1.22%组种蛋HR显著高于对照组(P<0.05),0.92%、1.02%组种蛋HU显著高于对照组(P<0.05),1.02%组种蛋ET显著高于对照组和0.82%、1.22%组(P<0.05)。饲粮精氨酸水平对产蛋期五龙鹅种蛋FR、QR、ESI、YC、AH和YR无显著影响(P>0.05)。
表2 饲粮精氨酸水平对产蛋期五龙鹅种蛋质量的影响

Table 2 Effects of dietary arginine level on breeding egg quality of Wulong geese during laying period

项目
Items
饲粮精氨酸水平 Dietary arginine level/% P
P-value
0.72(对照组
Control group)
0.82 0.92 1.02 1.12 1.22
孵化率 HR/% 87.30±1.44b 91.79±1.07a 92.49±0.28a 94.63±1.37a 92.27±0.56a 93.58±2.89a 0.042
受精率 FR/% 88.75±2.34 90.00±2.50 85.92±3.12 91.25±4.24 82.50±5.73 75.00±14.10 0.554
合格率 QR/% 96.25±5.59 97.50±5.59 96.08±3.58 98.75±2.79 98.75±2.79 96.25±8.38 0.911
蛋形指数 ESI 1.41±0.02 1.48±0.02 1.46±0.01 1.42±0.06 1.43±0.08 1.46±0.03 0.887
蛋黄颜色 YC 6.64±0.38 6.96±0.28 6.32±0.23 6.72±0.24 6.46±0.18 6.86±0.97 0.678
蛋白高度
AH/mm
14.16±0.55 14.87±0.11 15.02±0.22 14.64±0.15 14.04±0.41 14.31±0.18 0.197
哈氏单位 HU 110.71±2.61b 113.86±0.79ab 115.75±0.43a 116.04±0.29a 113.70±0.47ab 113.80±0.61ab 0.049
蛋壳厚度
ET/mm
0.52±0.03b 0.56±0.01b 0.58±0.01ab 0.60±0.02a 0.58±0.01ab 0.54±0.02b 0.018
蛋黄比率
YR/%
45.29±1.61 42.41±1.39 45.76±1.99 43.10±0.58 45.27±1.18 42.68±2.23 0.495

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

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

2.2 饲粮精氨酸水平对产蛋期五龙鹅血清激素含量的影响

表3可知,1.02%组血清INS含量显著高于对照组和1.12%、1.22%组(P<0.05),1.02%组血清P4含量显著高于对照组和0.82%、1.22%组(P<0.05)。饲粮精氨酸水平对产蛋期五龙鹅血清LH和FSH含量无显著影响(P>0.05)。
表3 饲粮精氨酸水平对产蛋期五龙鹅血清激素含量的影响

Table 3 Effects of dietary arginine level on serum hormone contents of Wulong geese during laying period

项目
Items
饲粮精氨酸水平 Dietary arginine level/% P
P-value
0.72(对照组
Control group)
0.82 0.92 1.02 1.12 1.22
胰岛素
INS/(mU/L)
9.41±0.71c 13.32±1.33ab 12.39±2.24abc 14.87±0.48a 10.82±0.94bc 10.00±0.61bc 0.030
促黄体生成素
LH/(mIU/mL)
23.37±0.46 24.06±0.95 24.25±0.50 24.31±1.47 26.72±0.63 26.43±1.24 0.102
促卵泡素
FSH/(mIU/mL)
14.85±0.23 14.99±0.76 16.55±0.46 17.14±0.61 16.62±0.52 15.68±0.85 0.066
孕酮
P4/(ng/mL)
7.74±0.27b 7.72±0.29b 8.27±0.22ab 8.94±0.25a 8.52±0.33ab 7.85±0.30b 0.025

2.3 饲粮精氨酸水平对产蛋期五龙鹅血清细胞因子含量的影响

表4可知,0.92%、1.02%组血清IL-1 含量显著低于对照组和1.12%、1.22%组(P<0.05),0.92%、1.02%组血清IL-4含量显著高于对照组和1.02%组(P<0.05)。饲粮精氨酸水平对产蛋期五龙鹅血清IL-2、IL-6和IL-10含量无显著影响(P>0.05)。
表4 饲粮精氨酸水平对产蛋期五龙鹅血清细胞因子含量的影响

Table 4 Effects of dietary arginine level on serum cytokine contents of Wulong geese during laying periodpg/mL

项目
Items
饲粮精氨酸水平 Dietary arginine level/% P
P-value
0.72(对照组
Control group)
0.82 0.92 1.02 1.12 1.22
白细胞介素-1β
IL-1β
282.40±15.86a 264.11±16.35ab 225.01±4.68b 229.98±13.93b 275.49±14.68a 275.83±5.66a 0.011
白细胞介素-2
IL-2
127.10±10.25 119.09±6.69 115.64±9.50 120.27±7.69 112.14±6.22 132.44±5.52 0.491
白细胞介素-6
IL-6
55.33±2.18 49.37±5.09 49.91±3.86 47.65±4.18 51.66±2.89 49.47±2.22 0.732
白细胞介素-4
IL-4
218.06±19.65c 318.86±36.35abc 384.18±56.15a 382.05±19.55a 342.55±34.30ab 245.15±36.89bc 0.012
白细胞介素-10
IL-10
113.24±10.70 118.99±15.26 133.01±10.77 146.20±12.34 141.95±8.34 136.20±8.81 0.289

3 讨论

3.1 饲粮精氨酸水平对产蛋期五龙鹅种蛋质量的影响

精氨酸及其代谢产物对动物繁殖性能有重要影响,补充精氨酸可以促进胎盘血管生成,改善胎盘功能[13]。Zhang等[14]研究发现,家鸽胚蛋注射1.14 mg精氨酸可显著提高HR,精氨酸调节能量代谢活动通过糖异生作用转化为葡萄糖,补充孵化过程所需的能量需求。Subramaniyan等[15]和Gao等[16]研究发现,肉鸡胚蛋注射适宜L-精氨酸可提高HR,卵内胚胎可利用精氨酸改善能量状态并调节肌肉蛋白质沉积,从而促进胚胎生长,提高HR。林昊[10]研究发现,0.96%和1.16%精氨酸水平组种鸡HR显著高于对照组。与上述试验结果一致,本研究发现0.82%、0.92%、1.02%、1.12%、1.22%组种蛋HR显著高于对照组,1.02%组HR最高。可能是母体内积累的精氨酸在种蛋中被胚胎利用,导致肌肉质量增加和胚胎沉重;也可能是精氨酸代谢产物NO可以调节卵泡发育、成熟等,影响HR[17]
ET和HU是反映种蛋质量、蛋白品质和新鲜度的重要蛋品质指标[18-20]。徐良梅等[21]研究发现,肉种鸡饲粮添加1.16%~1.76%精氨酸后,与对照组相比,种蛋HU有上升趋势,可能是精氨酸与其他氨基酸存在协同作用。本研究发现,与对照组相比,饲粮1.02%精氨酸水平可显著提高五龙鹅种蛋ET和HU。Xia等[22]研究发现,与对照组相比,饲粮1.46%精氨酸水平显著提高鸭蛋ET,与本研究结果一致。这可能是精氨酸对调节蛋壳矿物质沉积的禽类蛋壳基质特异性蛋白ovocleidin-17有促进作用,鹅蛋壳中的主要基质蛋白ansocalcin与鸡蛋壳基质特异性蛋白ovocleidin-17具有高度相似性,推测可能是精氨酸对ansocalcin有同样的促进作用[23-24]。也可能是精氨酸通过激活雷帕霉素靶蛋白通路调控蛋白质沉积,进而影响蛋品质。

3.2 饲粮精氨酸水平对产蛋期五龙鹅血清激素含量的影响

精氨酸可通过调节激素水平促进畜禽繁殖性能,INS能够调节糖代谢及脂肪合成的蛋白质激素,维持血糖动态平衡[25]。本研究发现,饲粮精氨酸水平对产蛋期五龙鹅血清INS含量有显著影响,且在精氨酸水平为1.02%时效果最佳。姚康等[26]研究表明,哺乳仔猪饲粮添加0.6%和0.8%精氨酸显著提高了血清INS含量。这可能是精氨酸通过代谢产生多胺和NO,影响了INS等内分泌激素的释放,改善了机体机能[27]。姜丽丽[28]研究发现,蛋雏鸭饲粮中添加0.2%精氨酸显著提高了血清INS含量。精氨酸刺激胰岛细胞后,通过本身携带的正电荷加强膜的去极化,增加钙离子流来促进INS的分泌,精氨酸也可通过抑制生长激素抑制素释放和增加垂体前叶细胞内钙离子流增加生长激素含量,刺激INS的释放[27,29]
生殖激素在卵泡发育成熟的主要调节机制下丘脑-垂体-卵巢轴中发挥了至关重要的作用[30]。P4由卵泡颗粒细胞分泌,通过调控多种类固醇激素促进卵泡发育和排卵[31]。FSH和LH协同作用与性腺,促进雌激素分泌,刺激卵泡生长,通过卵泡发育和增加排卵增加产蛋量[32]。有研究表明,妊娠母羊饲粮中添加0.5 g/kg过瘤胃精氨酸可显著提高血浆P4含量,促进排卵[33]。本研究发现,饲粮精氨酸水平对产蛋期五龙鹅血清P4含量有显著影响,且在精氨酸水平为1.02%时效果最佳,但对血清FSH和LH含量没有显著影响。NO在生殖系统中发挥作用,精氨酸是畜禽体内NO合成的唯一供氮前体[34],畜禽可以通过调控体内NO含量促进P4分泌[35],但仍需进一步的研究调查NO在生殖系统发育过程中的具体机制。

3.3 饲粮精氨酸水平对产蛋期五龙鹅血清细胞因子含量的影响

精氨酸可通过诱导型一氧化氮合酶催化生成免疫调节的细胞信号分子NO,阻止病菌侵害家禽,降低机体损伤[36-37]。细胞因子是一类能在细胞间传递信息并具有免疫调节功能的的蛋白质或小肽,参与炎症和免疫反应期间其他细胞和组织的激活和控制[38],IL-1β是一种促炎因子,在机体感染和损伤时启动炎症反应,促进清除病原体,保护机体免受侵害作用[39];IL-4是一种抗炎因子,具有多效应性免疫活性调节作用,抑制炎症性体液免疫反应[40]。张贝贝[41]研究发现,感染使肉鸡空肠IL-1β基因表达量升高,饲粮添加精氨酸可降低细胞因子的基因表达量。Gao等[42]研究发现,卵内注射10 g/L精氨酸溶液的肉鸡血清IL-4含量显著高于其他组,可能是精氨酸促进淋巴细胞增殖,调节IL-4的合成分泌,从而提高了肉鸡的免疫功能。本研究发现,与对照组相比,饲粮精氨酸水平为0.92%和1.02%时,可显著降低血清IL-1 含量,显著提高血清IL-4含量。推测可能是精氨酸通过产生NO降低促炎细胞因子的合成[43],调节免疫细胞因子合成和分泌[44],进而增强机体免疫功能。

4 结论

饲粮适宜精氨酸水平可提高五龙鹅种蛋质量,调节激素水平,增强机体免疫功能。在本试验条件下,饲粮适宜精氨酸水平为1.02%。
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