代乳粉水平对羔羊血液指标和肠道屏障功能的影响

doi:10.3864/j.issn.0578-1752.2020.02.015

摘要/Abstract

摘要：

【背景】代乳粉的合理利用对于提高羔羊健康和成活率具有重要意义,但代乳粉饲喂水平对羔羊免疫功能和肠道屏障功能的影响仍不清楚,羔羊免疫功能发育和肠道健康发育所需的最佳代乳粉饲喂水平仍无定论。【目的】从羔羊血液学指标动态变化规律、回肠和结肠组织形态、抗氧化指标、免疫指标以及肠道屏障功能相关基因表达量的角度分析代乳粉饲喂水平对羔羊免疫功能和肠道屏障功能的影响。【方法】选用16只初生重接近的初生湖羊公羔,随机分成两组,分别为对照组（CON,代乳粉每日饲喂量为羔羊体重的2%）和高饲喂水平组（H,代乳粉每日饲喂量为羔羊体重的4%）。所有羔羊从7日龄开始断母乳饲喂代乳粉,49日龄时屠宰。【结果】代乳粉饲喂水平对羔羊白细胞数、淋巴细胞数、中性粒细胞数、红细胞数和血红蛋白浓度影响不显著（P＞0.05）。21—49d羔羊淋巴细胞数、红细胞数、血红蛋白浓度随日龄逐渐增加,49日龄时显著较高（P＜0.05）,而中性粒细胞淋巴细胞比49日龄时显著较低（P＜0.05）。H组羔羊回肠绒毛高度显著高于CON组（P＜0.05）,回肠和结肠隐窝深度显著低于CON组（P＜0.05）。回肠和结肠丙二醛（MDA）含量和免疫球蛋白A（IgA）含量显著低于CON组（P＜0.05）。结肠Claudin4表达量显著低于CON组（P＜0.05）,回肠和结肠肿瘤坏死因子（TNF-α）表达量均显著低于CON组（P＜0.05）。【结论】高代乳粉饲喂水平可以降低肠道组织氧化损伤,促进肠绒毛发育,降低肠道物理屏障损伤进而减少肠道免疫反应。生产中可以通过提高早期羔羊代乳粉饲喂水平促进羔羊肠道健康发育。

关键词: 羔羊, 代乳粉, 血液学指标, 肠道屏障, 抗氧化

Abstract:

【Background】 The application of milk replacer powder has important significance for improving the health and survival rate of lambs. However, the effects of milk replacer feeding levels on immune function and intestinal barrier function of lambs were still unclear. The optimal level of milk replacer for development of immune function and intestinal health is still inconclusive. 【Objective】 The objective of this experiment was to analyze the effects of the milk replacer feeding level on the immune function and intestinal barrier function of lambs through the dynamic of hematological indicators, the morphology of ileum and colon, antioxidant index, immune index and expression of intestinal barrier function related genes.【Method】16 newborn lambs were randomly divided into two groups: the control group (CON, milk replacer feeding level was 2% of average body weight) and the intensive feeding group (H, milk replacer feeding level was 4% of their average body weight). All lambs were fed milk replacer from the 7th day of age and were slaughtered at 49 days of age. 【Result】The results showed that the milk replacer feeding level had no significant effects on leukocyte count, lymphocyte count, neutrophil count, red blood cell count and hemoglobin concentration (P＞0.05). The number of lymphocytes, red blood cells and hemoglobin in the 21-49d lambs increased gradually with age, significantly higher at 49 days of age (P＜0.05), while neutrophil lymphocytes were significantly lower than at 49 days (P＜0.05). The ileal villus height of the H group was significantly higher than that of the CON group (P＜0.05), the ileum and colon crypt depth was significantly lower than that of the CON group (P＜0.05), the ileal and colonic malondialdehyde (MDA) content and immunoglobulin A (The content of IgA) was significantly lower than that of CON group (P＜0.05). The expression of Claudin4 in colon was significantly lower than that in CON group (P＜0.05), and the expression of tumor necrosis factor (TNF-α) in ileum and colon was significantly lower than that in CON group (P＜0.05). 【Conclusion】The intensive milk replacer feeding could reduce intestinal oxidative damage, promote intestinal villus development, reduce intestinal physical barrier damage and reduce intestinal immune response. In the sheep production, an intensive milk replacer-feeding program benefits to the healthy intestinal development.

Key words: lamb, milk replacer, hematological index, intestinal barrier, antioxidant

李彦军,牛骁麟,张千,王国秀,李发弟,李飞,李冲,庞鑫,贾莉,樊海苗. 代乳粉水平对羔羊血液指标和肠道屏障功能的影响[J]. 中国农业科学, 2020, 53(2): 409-417.

LI YanJun,NIU XiaoLin,ZHANG Qian,WANG GuoXiu,LI FaDi,LI Fei,LI Chong,PANG Xin,JIA Li,FAN HaiMiao. Effects of Milk Replacer Feeding Level on Hematology Index and Gut Barrier Function in Lambs[J]. Scientia Agricultura Sinica, 2020, 53(2): 409-417.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2020.02.015

https://www.chinaagrisci.com/CN/Y2020/V53/I2/409

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参考文献 24

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原料 Ingredients	配比Proportion (%)	营养成分Chemical composition
苜蓿草粉 Alfalfa meal	18.50	干物质 DM (%)	90.96
玉米 Corn	21.00	消化能 DE (MJ·kg^-1)	13.01
膨化玉米 Extruded corn	22.30	粗蛋白质 CP (%)	19.50
麸皮 Bran	6.00	钙 Ca(%)	0.66
豆粕 Soybean meal	21.50	磷 P(%)	0.31
膨化大豆 Extruded-soybean	4.00	中性洗涤纤维 NDF（%）	18.87
玉米蛋白粉 Corn gluten meal	5.00
石粉 Limestone	0.30
预混料 Premix	1.00
食盐 Salt	0.40
合计 Total	100.00

基因 Gene	检索号 Gene bank number	引物序列 Primer sequence	片段长度 Size (bp)	扩增效率 Amplification efficiency
Claudin-1	NM_001185016.1	F: 5' AATACATTGAGGTCACCGAGTA 3'	191	98.93%
		R: 5' GATTAGGCAAGGAAAGGCAC 3'
Claudin-4	NM_001185017.1	F: 5' GCCTTCATCGGCAGCAACAT 3'	115	92.75%
		R: 5' CCAGCAGCGAGTCGTACACCTT 3'
Occludin	XM_012145891.2	F: 5' AGTGGTAACTTGGAGACGCTTTC 3'	107	95.17%
		R: 5' CCTCCCGTCGTGTAGTCTGTT 3'
TLR4	NM_001135930.1	F: 5' GGTTTCAGGAATGCCACTT 3'	142	98.14%
		R: 5' CTTTCACCTCTGCCATACTTT 3'
TNFα	NM_001114186.1	F: 5' ACGGCGTGGAGCTGAAA 3'	132	92.18%
		R: 5' CTGATGGTGTGGGTGAGGAA 3'
β-Actin	NM_001009784.2	F: 5' TCCGTGACATCAAGGAGAAGC 3'	266	91.82%
		R: 5' CCGTGTTGGCGTAGAGGT 3'

项目 Items	处理组 Treatment	日龄（d）					标准误 SEM	P值 P value
项目 Items	处理组 Treatment	21	28	35	42	49	标准误 SEM	饲喂水平 Treatment	日龄 Age	饲喂水平×日龄 Treatment × Age
白细胞 Leukocyte (×10⁹cells/L)	CON	8.28	8.31	9.20	8.28	9.86	0.297	0.839	0.615	0.954
白细胞 Leukocyte (×10⁹cells/L)	H	8.33	8.26	8.34	8.91	9.48	0.297	0.839	0.615	0.954
淋巴细胞 Lymphocyte (×10⁹cells/L)	CON	3.52b	3.32b	4.01ab	4.32ab	5.47a	0.153	0.763	<0.001	0.966
淋巴细胞 Lymphocyte (×10⁹cells/L)	H	3.57b	3.57b	3.86ab	4.32ab	5.88a	0.153	0.763	<0.001	0.966
中性粒细胞 Neutrophil (×10⁹cells/L)	CON	3.49	3.69	3.68	2.75	3.30	0.200	0.592	0.738	0.908
中性粒细胞 Neutrophil (×10⁹cells/L)	H	3.36	3.37	3.44	3.25	2.40	0.200	0.592	0.738	0.908
中性粒细胞/淋巴细胞NLR	CON	1.02	1.12	1.03	0.66	0.61	0.062	0.813	0.034	0.972
中性粒细胞/淋巴细胞NLR	H	1.05a	1.04a	0.99ab	0.78ab	0.44b	0.062	0.813	0.034	0.972
红细胞 Red blood cells (×10¹²cells/L)	CON	8.47ab	7.91b	7.94b	8.37ab	9.22a	0.080	0.920	<0.001	0.696
红细胞 Red blood cells (×10¹²cells/L)	H	8.11b	7.80b	8.22b	8.45b	9.40a	0.080	0.920	<0.001	0.696
血红蛋白浓度Hemoglobin concentration (g·L^-1)	CON	122.64ab	109.79b	111.94b	116.67ab	127.43a	1.346	0.705	0.001	0.698
血红蛋白浓度Hemoglobin concentration (g·L^-1)	H	114.72ab	108.71b	115.00ab	116.43ab	127.92a	1.346	0.705	0.001	0.698

	项目Item	CON	H	P值 P value
回肠 Ileum	肠绒毛高度Villus height (μm)	349.58±39.01	429.77±62.51	0.041
回肠 Ileum	肠绒毛宽度Villus width (μm)	104.43±23.06	130.49±37.13	0.219
	隐窝深度Crypt depth(μm)	158.63±28.27	114.46±15.71	0.027
	肌层厚度Muscle layer thickness (μm)	173.06±47.28	175.40±15.44	0.919
结肠 Colon	肠绒毛高度Villus height (μm)	371.99±44.80	350.01±74.85	0.544
结肠 Colon	肠绒毛宽度Villus width (μm)	86.79±3.98	75.80±23.36	0.544
	隐窝深度Crypt depth (μm)	190.09±52.96	135.58±27.37	0.041
	肌层厚度Muscle layer thickness (μm)	167.97±39.30	164.22±31.93	0.859

	项目Item	CON	H	P值P value
回肠 Ileum	MDA（nmol·g^-1）	14.02±5.00	6.59±3.14	0.029
回肠 Ileum	GSH（U·g^-1）	448.72±75.27	452.33±77.20	0.936
	SOD（U·g^-1）	1236.01±203.75	1246.15±117.87	0.929
	IgA（mg·g^-1）	0.37±0.06	0.15±0.09	0.005
结肠 Colon	MDA（nmol·g^-1）	61.49±14.56	26.03±13.84	0.003
结肠 Colon	GSH（U·g^-1）	405.04±102.68	396.40±105.56	0.907
	SOD（U·g^-1）	1134.25±218.65	932.68±141.25	0.111
	IgA（mg·g^-1）	0.90±0.21	0.56±0.23	0.015