Effects of Allicin and Bacillus natto Supplements on Production, Antioxidant Enzymes, Intestinal Morphology, and Cecal Microbiota of Fujian White Rabbits
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摘要:
目的 研究大蒜素、纳豆菌及其复合物对福建白兔生产性能、抗氧化活性、肠道形态和菌群的影响，筛选合适的添加剂替代饲料抗生素。 方法 选取48只体重相近的40日龄福建白兔，随机分成4组，每组12只（公母各半），试验期50 d。对照组饲喂基础饲粮（A组，CK），试验在基础饲粮上分别添加200 mg·kg−1大蒜素、0.5%纳豆菌、200 mg·kg−1大蒜素+0.5%纳豆菌，设置大蒜素处理组（B组）、纳豆菌处理组（C组）和大蒜素+纳豆菌处理组（D组）。试验期间测定生产性能，试验结束测定屠宰性能、血清抗氧化酶、肠道形态和盲肠菌群结构。 结果 （1）试验组的腹泻率和死亡率都低于对照组；平均日增重大蒜素+纳豆菌处理（D组）极显著高于基础饲粮处理组（A组）和大蒜素处理组（B组）（P＜0.01），显著高于纳豆菌处理组（C组）（P＜0.05）；D组料重比最低，半净膛重极显著高于A组和B组（P＜0.01）；全净膛率和半净膛率各组之间差异不显著。（2）血清中GSH-Px含量，对照组均显著低于试验组，且A组和B组差异显著（P＜0.05）， A组和C、D组差异极显著（P＜0.01）。（3）回肠隐窝深度B组显著高于C组（P＜0.05）；空肠隐窝深度，对照组低于试验组，且与B组差异极显著（P＜0.01），与D组差异显著（P＜0.05）。（4）菌群结构分析，科水平和属水平盲肠unclassified_o__Clostridiales菌科、菌属在组间差异均显著 （P＜0.05），且A组显著高于D 组；属水平Ruminococcaceae_UCG-010在组间差异显著 （P＜0.05），A组显著高于C 组。 结论 大蒜素和纳豆菌联合使用可提高福建白兔平均日增重、半净膛重，降低料重比，增加空肠隐窝深度，减少有害菌，调整肠道菌群平衡，降低腹泻和死亡，促进生长发育。 Abstract:Objective Effects of addition of allicin and/or Bacillus natto in feed on the production performance, antioxidant enzymes, intestinal morphology, and cecal microbiota of Fujian white rabbits were studied for forage supplement as an antibiotic replacement. Method Forty-eight healthy 40-day-old Fujian white rabbits with similar body weight were randomly divided into 4 groups of 12 rabbits each, half male and half female, for a 50 d feeding trial. The rabbits in the control group were fed a basal diet (Group A), while those in the treatment groups the basal diet supplemented with 200 mg·kg−1 allicin (Group B), 0.5% B. natto (Group C), and 200 mg·kg−1 allicin+0.5% B. natto (Group D). Growth of the rabbits was monitored during the trial, and at the end, the animals were slaughtered to determine the meat production, serum antioxidant enzymes, intestinal morphology, and cecum microbiota. Results (1) The rates of diarrhea occurrence and mortality of the rabbits in Group B, C, and D were lower than those of Group A. The average daily weight gain of Group D was significantly higher than that of Group A and B at P＜0.01 and Group C at P＜0.05. Group D had the lowest F/G among all, and significantly higher half-eviscerated weight than that of Group A or Group B (P＜0.01). No significant differences were observed in the eviscerated and half-eviscerated rates among the groups. (2) On serum GSH-Px, Group A was significantly lower than Group B, C and D. The difference between group A and Group B was significant (P＜0.05), while those between Group A and Group C and between Group A and Group D extremely significant (P＜0.01). (3) The ileal crypt depth of Group B was significantly higher than Group C(P ＜0.05). Group A had a lower depth of jejunum crypt than the treatment groups and significantly different from Group B (P＜0.01) and Group D (P＜0.05). (4) The unclassified_o__Clostridiales significantly differed at both family and genus levels among the groups (P＜0.05) with Group A being significantly higher than Group D. Whereas, at genus level, Ruminococcaceae_UCG-010 significantly differed among the groups (P＜0.05), but Group A was significantly higher than Group C. Conclusion The combined use of allicin and B. natto in feed significantly increased the average daily weight gain and half-eviscerated weight, decreased the F/G, increased the depth of jejunum crypt, reduced the harmful bacteria population, balanced the cecum microbiota, mitigated diarrhea and deaths, promoted the growth and development of Fujian white rabbits. -
Key words:
- Fujian white rabbit /
- allicin /
- Bacillus natto /
- production performance /
- intestinal morphology /
- cecal microbiota
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图 1 福建白兔肠道组织切片形态（100×）
Figure 1. Morphology of sectioned intestinal tissues of Fujian white rabbits (100×)
图 2 不同处理组门水平肠道微生物丰度
Figure 2. Intestinal microbial abundance at phylum level of treatment groups
图 3 不同处理组肠道微生物科水平丰度
Figure 3. Intestinal microbial abundance at family level of treatment groups
图 4 不同处理组肠道微生物科水平菌群差异性
Figure 4. Differential intestinal microflora at family level of treatment groups
图 5 不同处理组肠道微生物属水平丰度
Figure 5. Intestinal microbial abundance at genus level of treatment groups
图 6 不同处理组肠道微生物菌群属水平差异性
Figure 6. Differential intestinal microflora of treatment groups
表 1 基础饲粮组成及营养水平（干物质基础）
Table 1. Composition and nutrient levels of basic diets (DM basis)
项目
Items含量
Content原料 Ingredients 玉米 Corn/% 30.0 豆粕 Soybean meal/% 15.0 麸皮 Wheat bran/% 25.0 花生壳粉 Peanut shell powder/% 5.0 草粉 Grass meal/% 20.0 预混料 Premix1）/% 5.0 合计 Total/% 100.0 营养水平 Nutrient levels2） 粗蛋白质 Crude protein/% 17.0 消化能 Digestible energy/（MJ·kg−1） 10.6 粗纤维 Crude fiber/% 13.0 钙 Ca/% 1.0 磷 P/% 0.5 注：1）预混料为每千克饲粮提供：赖氨酸1 500 mg，蛋氨酸 1 500 mg，CaHPO4 15 000 mg，NaCl 5 000 mg，VA 10 000 IU，VD3 2 000 IU，VE 50 mg，VK3 2.5 mg，硫胺素5 mg，VB12 1 mg，核黄素10 mg，泛酸50 mg，烟酸20 mg，叶酸2.5 mg，氯化胆碱400 mg，Fe 100 mg，Zn 50 mg，Cu 40 mg，Mn 30 mg，I 0.5 mg，Se 0.05 mg，其余为载体补足。2）营养水平为计算值。
Note: 1) Premix provided the following per kg of diets: Lys 1 500 mg, Met 1 500 mg, CaHPO415 000 mg, NaCl 5 000 mg, VA 10 000 IU, VD3 2 000 IU, VE 50 mg, VK3 2.5 mg, thiamine 5 mg, VB12 1 mg, riboflavin 10 mg, pantothenic acid 50 mg, nicotinic acid 20 mg, folic acid 2.5 mg, choline chloride 400 mg, Fe 100 mg, Zn 50 mg, Cu 40 mg, Mn 30 mg, I 0.5 mg, Se 0.05 mg, the rest was meal carrier complement. 2) Nutrient levels were calculated.
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表 2 不同添加剂对福建白兔腹泻率和死亡率的影响
Table 2. Effects of supplements on rates of diarrhea incident and mortality of Fujian white rabbits
组别
Groups腹泻率 Number of diarrhea/% 死亡率 Mortality/% 平均
Average40～60 d 60～90 d 平均
Average40～60 d 60～90 d 对照A组 CK 50.00 33.33 25.00 33.33 33.33 0.00 处理B组 Treatment B 25.00 25.00 0.00 33.33 33.33 0.00 处理C组 Treatment C 25.00 25.00 0.00 25.00 16.67 8.33 处理D组 Treatment D 8.33 8.33 0.00 16.67 8.33 8.33
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表 3 不同添加剂对福建白兔生长性能的影响
Table 3. Effect of supplements on growth performance of Fujian white rabbits
组别
Groups初始体重
Original weight/g终末体重
Final weight/g平均日采食量
Average feed intake/（g·d−1）平均日增重
Average weight gain/（g·d−1）料重比
F/G处理A组 CK 667.25±78.68 1635.31±131.67 ABb 110.26±8.70 ABab 19.36±2.16 Bb 5.74±0.54 处理B组 Treatment B 659.13±86.17 1622.25±98.21 Bb 105.87±9.75 Bb 19.43±2.07 Bb 5.81±0.86 处理C组 Treatment C 690.33±83.40 1697.61±108.44 ABab 118.20±7.68 Aa 20.27±1.94 ABb 5.92±0.70 处理D组 Treatment D 618.9±112.43 1772.00±90.22 Aa 119.92±8.23 Aa 23.06±3.10 Aa 5.35±0.68 注：同列数据后字母相同或不标者表示差异不显著（P＞0. 05）；小写字母不同表示差异显著（P＜0.05）； 大写字母不同表示差异极显著（P＜0.01）。表4～8同。
Note: The datas with the same or no letters in a line mean no significance (P＞0. 05), the datas with different small letters indicate significance difference (P＜0.05), the datas with different capital letters indicate extremely significances difference (P＜0.01). Same for Table 4–8.
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表 4 不同添加剂对福建白兔屠宰性能的影响
Table 4. Effect of supplements on slaughter performance of Fujian white rabbits
组别
Groups全净膛重
Carcass net weight/g半净膛重
Half eviscerated weight/g全净膛率
Whole net carcass rate/%半净膛率
Half-complete ratio/%对照A组 CK 799.06±65.05 ABb 880.31±68.27 Bb 0.49±0.02 0.54±0.02 处理B组Treatment B 790.63±44.17 Bb 870.56±49.53 Bb 0.49±0.02 0.54±0.02 处理C组Treatment C 835.39±47.79 ABab 928.78±52.59 ABa 0.49±0.03 0.55±0.03 处理D组Treatment D 861.65±56.93 Aa 957.8±56.53 Aa 0.49±0.02 0.54±0.02
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表 5 不同添加剂对福建白兔血清抗氧化酶的影响
Table 5. Effect of supplements on serum antioxidant enzymes of Fujian white rabbits
组别
Groups总抗氧化能力
T-AOC/（mmol·L−1）谷胱甘肽过氧化物酶
GSH-Px/（U·mL−1）丙二醛
MDA/（nmol·mL−1）对照A组 CK 0.59±0.08 202.03±37.00 Bb 5.23±1.14 处理B组 Treatment B 0.61±0.08 275.63±53.97 ABa 5.38±1.51 处理C组 Treatment C 0.58±0.12 311.51±49.82 Aa 4.59±2.52 处理D组 Treatment D 0.56±0.06 318.22±69.78 Aa 4.43±1.29
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表 6 不同添加剂对福建白兔肠道绒毛长度和隐窝深度的影响
Table 6. Effects of supplements on intestinal villus length and crypt depth of Fujian white rabbits
（单位：mm） 组别
Groups绒毛长度 Villus length 隐窝深度 Crypt depth 十二指肠
Duodenum回肠
Ileal空肠
Jejunum十二指肠
Duodenum回肠
Ileal空肠
Jejunum对照A组 CK 0.68±0.07 0.54±0.13 0.51±0.08 0.13±0.02 0.10±0.02 Aab 0.10±0.01 Bc 处理B组 Treatment B 0.62±0.13 0.49±0.09 0.49±0.05 0.13±0.05 0.12±0.04 Aa 0.13±0.03 Aa 处理C组 Treatment C 0.65±0.07 0.57±0.07 0.53±0.05 0.10±0.02 0.09±0.01 Ab 0.11±0.02 ABb 处理D组 Treatment D 0.62±0.13 0.52±0.06 0.52±0.07 0.12±0.03 0.10±0.02 Aab 0.12±0.03 ABab
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表 7 不同添加剂对福建白兔肠道绒毛数量和隐窝数量的影响
Table 7. Effects of supplements on intestinal villi population and crypts of Fujian white rabbits
（单位：根·mm−2） 组别
Groups绒毛数量 Villus number 隐窝数量 Crypt number 十二指肠
Duodenum回肠
Ileal空肠
Jejunum十二指肠
Duodenum回肠
Ileal空肠
Jejunum对照A组 CK 4.15±0.88 7.62±1.74 7.44±2.68 13.08±4.03 18.01±7.88Aa 23.5±17.38 处理B组 Treatment B 4.75±1.17 7.70±0.96 6.73±1.02 18.01±6.76 17.34±4.75Aab 28.94±27.38 处理C组 Treatment C 4.58±1.05 7.61±1.05 6.28±0.60 14.68±6.50 12.19±3.48Ab 16.54±6.34 处理D组 Treatment D 4.75±0.95 7.41±1.87 6.16±0.76 14.29±4.78 13.75±2.62Ab 14.32±4.48
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表 8 试验各组测序结果及Alpha多样性分析
Table 8. Sequence and alpha diversity of treatment groups
项目
Item对照A组
CK处理B组
Treatment B处理C组
Treatment C处理D组
Treatment D分类单元数量 OUT/个 648.00±49.88 571.25±90.6 578.56±144.44 575.20±147.72 覆盖率 Coverage/% 99.58±0.03 99.63±0.05 99.60±0.11 99.62±0.09 Shannon指数 Shannon index 4.82±0.18 Aa 4.59±0.36 Aab 4.53±0.33 Ab 4.54±0.52 Aab Simpson指数 Simpson index 0.02±0.01 0.03±0.02 0.04±0.02 0.03±0.02 ACE指数 ACE index 742.16±54.21 657.55±98.67 673.62±165.4 668.17±150.36 Chaol指数 Chaol index 751.06±49.72 668.86±98.81 689.77±171.52 673.87±169.42 注：同行数据后字母相同或不标者表示差异不显著（P＞0. 05）；小写字母不同表示差异显著（P＜0.05）；大写字母不同表示差异极显著（P＜0.01）。
Note: The datas with the same or no letters indicate no significance in a row(P＞0.05), the datas with different small letters indicate significance difference(P＜0.05), the datas with different capital letters indicate extremely significances difference(P＜0.01).
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