Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (2): 422-434.doi: 10.3864/j.issn.0578-1752.2021.02.017

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Comparison of Growth Physiology and Gut Microbiota Between Healthy and Diarrheic Lambs in Pre- and Post-Weaning Period

YANG NingZhi1(),LI Ting1,WANG Yan1,CHEN Zhuo1,MA YiCheng1,REN QiangLin1,LIU JiaJia2,YANG HuiGuo2,YAO Gang1()   

  1. 1College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052
    2Institute of Animal Science, Academy of Animal Science and Veterinary Medicine, Urumqi 830026
  • Received:2020-03-08 Accepted:2020-07-10 Online:2021-01-16 Published:2021-02-03
  • Contact: Gang YAO E-mail:yangningzhi1@126.com;yg@xjau.edu.cn

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Abstract:

【Objective】Lamb diarrhea is one of the common clinical diseases caused by the certain pathogens or the interaction with malnutrition, abnormal management, and environmental stress during newborn and weaning periods, which hazards the lamb production. There is lack of enough investigation about the differences of lambs' physio-biochemistry with their gut microbiota caused by diarrhea between suckling and weaned periods. Therefore, this study was intended to compare the growth physiology with gut microbiota of diarrheic lambs in pre-and post-weaning period, so as to lay a scientific foundation for prevention and treatment of lamb diarrhea occurred at different growth stages precisely and effectively. 【Method】10 suckling healthy lambs and 10 diarrheic lambs at age of 31-45 day, and another 10 weaned healthy lambs and 10 diarrheic lambs at age of 60-75 day were selected and paired to check the growth status, blood physio-biochemistry and inflammatory factors. The rectal fecal samples were used to analyze gut microbiota composition and structure using 16S rRNA. 【Result】1. There was no significant difference in body weight between healthy and diarrhea lambs before weaning, but after weaning, the body weight of diarrheic lambs was extremely lower than that of healthy lambs (P<0.01), and the body temperature, pulse and respiratory rate of diarrheic lambs were also significantly lower than those of healthy lambs (P<0.05). 2. Before weaning, the total protein, globulin, and cholesterol in diarrheic lambs were significantly lower than those in healthy lambs (P<0.05). After weaning, the total number of leukocytes in diarrheic lambs was extremely higher than that of healthy lambs (P<0.01). And the blood urea, nitrogen, creatinine, phosphorus, glucose and total bilirubin in diarrheic lambs were significantly lower than those in healthy lambs (P<0.05). After weaning, IL-4, IL-6 and IL-8 in diarrheic lambs were significantly higher than those in healthy lambs (P<0.05). 3. The percentage of common OTUs in the gut of healthy and diarrheic lambs after weaning was almost as doubled as that before weaning. Alpha diversity of the gut microbiota in diarrheic lambs before weaning was significantly reduced as compared to their healthy counterpart (P<0.05). And Beta diversity of the gut microbiota weighted PCoA in diarrheic lambs before weaning was significantly different from that in the healthy lambs (Weighted UniFrac Adonis and Anosim tests, P<0.05). At phylum level: before weaning, the relative abundance of Fusobacteria was significantly increased (P<0.05), whereas that of Verrucomicrobia and Actinobacteria was significantly decreased in the gut of diarrheic lambs as compared to their healthy counterpart (P<0.05). After weaning, the relative abundance of Lentisphaerae and Actinobacteria was significantly reduced as compared to their healthy counterpart (P<0.05). At genus level: before weaning, the relative abundance of Fusobacterium and Clostridium were significantly increased, while Akkermansia, Desulfovibrio and Butyricimonas were significantly decreased in the gut of diarrheic lambs as compared to their healthy counterpart (P<0.05). After weaning, the relative abundance of Ruminococcus was significantly decreased, while Desulfovibrio significantly was increased in the gut of diarrheic lambs as compared to their healthy counterpart (P<0.05). 【Conclusion】It was concluded that before weaning the content of blood proteins and the blood fat decreased significantly, whereas after weaning, the elevation of white blood cells and the decrease of CREA, UREA and the blood glucose was significant in diarrheic lambs. Before weaning, the more obvious gut microbiota dysbiosis was observed, whereas after weaning more significant gut inflammatory activity was observed in diarrheic lambs. The ration structure and the environment during weaning time could be the crucial factors for shifting aforementioned differences.

Key words: lamb, weaning, diarrhea, physio-biochemical and inflammatory factor, gut microbiota

Table 1

Experimental grouping"

分组
Grouping		 日龄（天）
Age(d)		 健康组
Health group		 腹泻组
Diarrhea group
断奶前（Suckling） 31-45 H组（Health） D组（Diarrhea）
断奶后（Weaned） 60-75 h组（health） d组（diarrhea）

Table 2

Diet composition of lambs pre- and post- weaning period"

原料
Ingredient		 日粮组成 Diet composition (%) 营养成分
Nutritional components		 营养水平 Nutrient levels
断奶前Suckling 断奶后Weaned 断奶前Suckling 断奶后Weaned
玉米Corn 43.12 25.20 干物质 DM (%) 82.29 86.65
麸皮Wheat bran 3.75 4.40 消化能DE (MJ·kg-1) 10.41 6.43
豆粕Sybean meal 16.50 4.40 粗蛋白 CP (%) 16.76 10.76
棉粕Cottonseed meal 3.00 1.20 粗纤维 CF (%) 11.01 24.48
葵粕Sunflower meal 2.63 2.00 粗灰分Ash (%) 4.43 6.27
酵母Brewers dried yeast 0.70 1.60 钙 Ca (%) 0.57 0.51
食盐NaCl 0.80 0.40 总磷 P (%) 0.34 0.23
石粉Limestone 0.80 0.40
碳酸氢钠Sodium bicarbonate 3.70 0.40
麦秸Wheat straw 10.00 40.00
苜蓿草粉Alfalfa meal 14.00 19.00
复合预混料Premix 1.00 1.00
合计Total 100.00 100.00

Fig. 1

Fecal consistency score（left-right：score of 0, 1, 2, 3 and 4）"

Table 3

Ratios of diarrhea and death of diarrheic lambs in pre- and poster- weaning time"

日龄
Age (d)		 总头数
N. lambs		 腹泻只（次）数
N. diarrhea		 腹泻率
Diarrhea rate		 死亡数
N. death		 病死率
Mortality
1-15 80 34 2.83 18 52.90
16-30 122 55 3.01 12 21.80
31-45 59 22 2.49 9 40.90
60-75 200 68 2.27 14 20.60
总计Total 461 179 2.59 53 29.60

Table 4

Blood physiological parameters of healthy and diarrheic lambs in pre- and poster- weaning time"

指标
Index		 断奶前Suckling 断奶后Weaned
健康Health 腹泻Diarrhea 健康Health 腹泻Diarrhea
白细胞WBC 8.69±0.39 9.12±0.45 8.96±0.36A 11.20±0.47B
淋巴细胞LY 4.42±0.32 5.00±0.26 5.84±0.34 6.23±0.42
中性粒细胞Gran 4.14±0.46 4.33±0.85 5.36±0.87 5.29±0.72
血红蛋白HGB 107.00±3.78 108.00±5.16 127.00±4.44 141.00±5.06
红细胞RBC 9.43±0.24 8.97±0.43 8.69±0.24 9.30±0.34
血小板PLT 507.00±38.9 487.00±38.40 750.00±40.00 768.00±55.00

Table 5

Blood biochemical parameters of healthy and diarrheic lambs in pre- and poster- weaning time"

指标
Index		 断奶前Suckling 断奶后Weaned
健康Health 腹泻Diarrhea 健康health 腹泻diarrhea
碱性磷酸酶ALKP(U·L-1) 355.00±90.30 169.20±73.13 38.80±9.20 69.80±38.27
丙氨酸转移酶ALT(U·L-1) 186.40±152.42 228.00±258.80 37.20±16.77 37.40±18.77
胰淀粉酶AMYL(U·L-1) 64.60±12.88 59.20±34.78 32.00±30.54 54.80±30.26
白蛋白ALB(g·L-1) 19.00±2.35 19.60±1.82 15.20±5.72 19.80±6.61
球蛋白GLOB(g·L-1) 48.80±3.97a 35.60±5.35b 56.80±12.48 47.40±14.08
总蛋白TP(g·L-1) 66.40±15.79a 55.00±5.24b 72.00±9.05 67.20±9.41
胆固醇CHOL(mmol·L-1) 3.70±1.87a 2.52±0.86b 0.58±0.34 0.77±0.22
肌酐CREA(mmol·L-1) 39.00±14.07 27.20±5.72 35.80±7.60A 22.90±5.60B
尿素氮UREA(mmol·L-1) 4.18±1.45 3.64±1.28 9.15±2.50A 6.48±0.94B
总胆红素BIL(mmol·L-1) 2.80±1.64 2.00±2.23 4.00±2.94a 1.20±0.44b
血糖GLU(mmol·L-1) 5.36±0.71 5.42±1.72 4.85±1.01a 3.93±0.64b
钙离子Ca(mmol·L-1) 2.50±0.08 2.35±0.20 2.14±0.27 2.11±0.10
磷PHOS(mmol·L-1) 2.89±0.82a 2.18±0.40b 2.52±0.58A 1.64±0.31B

Table 6

Significance test of bacterial flora difference"

方法
Method name		 P 断奶前
Suckling		 断奶后
Weaned
加权UniFrac
Weighted UniFrac		 Adonis Pr (>F) 0.02 0.26
Anosim P-value 0.04 0.15

Fig. 2

Body weight, temperature, pulse and respiratory parameters of healthy and diarrheic lambs in pre- and poster- weaning time"

Fig. 3

Changes of inflammatory factors of healthy and diarrheic lambs in pre- and poster- weaning time"

Fig. 4

Venn analysis of OTUs of gut microbiota of healthy and diarrheic lambs in pre- and poster- weaning time"

Fig. 5

Alpha-diversity index of the gut microbiota of healthy and diarrheic lambs in pre- and poster- weaning time"

Fig. 6

Weighted UniFrac PCA of gut microbiota of healthy and diarrheic lambs in pre- and poster- weaning time"

Table 7

Number of OTUs of gut microbiota at each classification level"

分类水平
Classification level		 断奶前 Suckling 断奶后 Weaned
健康 Health 腹泻 Diarrhea 健康 Health 腹泻 Diarrhea
门Phylum 8.70±1.25 8.00±2.67 10.70±2.50 11.30±1.59
纲Class 16.00±2.70 14.00±3.68 20.80±3.67 20.40±2.11
目Order 20.10±4.30 16.00±4.97 27.00±5.56 27.10±4.04
科Family 35.3±7.13 28.80±7.91 45.90±9.37 46.30±5.53
属Genus 56.1±9.75a 46.40±12.07b 67.70±13.33 67.80±8.53
种Species 63.3±9.25a 52.70±12.27b 69.30±13.49 70.10±9.27

Fig. 7

Relative abundance and difference of gut microbiota of healthy and diarrheic lambs in pre- and poster- weaning time at phyla level"

Fig. 8

Relative abundance and difference of gut microbiota of healthy and diarrheic lambs in pre- and poster- weaning time at genera level"

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