Effects of Dietary Adding Different Probiotics on Growth Performance, Slaughter Performance, Ileum Mucosal Immune Indexes and Cecal Fermentation Indexes of Meat Rabbits

LIU Gongyan; GAO Shuxia; BAI Liya; LIU Man; LI Mingyong; YANG Liping; JIANG Wenxue; WANG Peng; SUN Haitao

doi:10.3969/j.issn.1006-267x.2023.01.050

2023 , Vol. 35 >Issue 1: 527 - 535

DOI: https://doi.org/10.3969/j.issn.1006-267x.2023.01.050

RESEARCH PAPER

Effects of Dietary Adding Different Probiotics on Growth Performance, Slaughter Performance, Ileum Mucosal Immune Indexes and Cecal Fermentation Indexes of Meat Rabbits

LIU Gongyan ^,¹ ,
GAO Shuxia ¹ ,
BAI Liya ¹ ,
LIU Man ² ,
LI Mingyong ² ,
YANG Liping ¹ ,
JIANG Wenxue ¹ ,
WANG Peng ³ ,
SUN Haitao ^,¹^,^*

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¹ Key Laboratory of Livestock and Poultry Multiomics of Ministry of Agriculture and Rural Affairs, Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Husbandry and Veterinary, Shandong Academy of Agricultural Sciences, Jinan 250100, China
² Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Qingdao Kangda Rabbit Industry Development Co., Ltd., Qingdao 266000, China
³ Shandong Zhengyu Rabbit Industry Co., Ltd., Linyi 273400, China

^* associate professor, E-mail: wwww8888@163.com

Received date: 2022-05-20

Online published: 2023-01-11

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Abstract

This experiment was conducted to study the effects of dietary adding different probiotics on growth performance, slaughter performance, ileum mucosal immune indexes and cecal fermentation indexes of meat rabbits, so as to provide theoretical reference for application of probiotics in rabbit production. A total of 400 commercial Ira meat rabbits with 28-day-old weaned, similar weight and good health were randomly divided into 4 groups with 10 replicates in each group and 10 rabbits in each replicate. Rabbits in the control group was fed a basal diet, while others in experimental groups were fed basal diets added 10⁸ CFU/kg Lactiplantbacillus plantarum, Pediococcus pentosaceus and Clostridium butyricum, respectively. The pre-experimental period lasted for 7 days, and the formal experimental period lasted for 40 days. The results showed as follows: 1) dietary adding different probiotics had no significant effects on growth performance, slaughter performance and visceral organ development indexes of meat rabbits (P>0.05). Dietary adding different probiotics had a trend to decrease the diarrhea rate of meat rabbits (P=0.059 5), and had a trend to increase the survival rate of meat rabbits (P=0.060 3). 2) The jejunum villi height of experimental groups was significantly higher than that of the control group (P<0.05), and the jejunum crypt depth of Clostridium butyricum group was significantly higher than the other groups (P<0.05). 3) The ileum secretory immunoglobulin A (sIgA) content of experimental groups was significantly higher than that of the control group (P<0.05), and the contents of tumour necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in ileum of experimental groups was significantly lower than those of the control group (P<0.05). 4) The cecal contents pH of experimental groups was significantly lower than that of the control group (P<0.05), the contents of acetic acid and total volatile fatty acids in cecal contents of experimental groups was significantly higher than those of the control group (P<0.05), and the cecal contents ammonia nitrogen content of Clostridium butyricum group was significantly lower than that of the control group (P<0.05). In conclusion, dietary adding different probiotics can improve the intestinal morphology and structure, change the composition of volatile fatty acids in cecum contents, improve the immune performance of intestinal mucosa, and promote the intestinal health of meat rabbits. Based on the results of this experiment, Clostridium butyricum has a good effect on maintain the intestinal morphology and structure of meat rabbits.

Key words： Lactiplantbacillus plantarum; Pediococcus pentosaceus; Clostridium butyricum; meat rabbits; growth performance; immune performance; cecal fermentation

Cite this article

LIU Gongyan , GAO Shuxia , BAI Liya , LIU Man , LI Mingyong , YANG Liping , JIANG Wenxue , WANG Peng , SUN Haitao . Effects of Dietary Adding Different Probiotics on Growth Performance, Slaughter Performance, Ileum Mucosal Immune Indexes and Cecal Fermentation Indexes of Meat Rabbits[J]. Chinese Journal of Animal Nutrition, 2023 , 35(1) : 527 -535 . DOI: 10.3969/j.issn.1006-267x.2023.01.050

动物胃肠道内存在大量微生物,在机体营养消化吸收、维持肠道屏障结构完整性和抵御病原体方面发挥着重要作用。胃肠道微生物菌群失调不仅会导致消化功能紊乱,还会引起多种肠道外疾病^[1⇓-3]。益生菌最初被定义为“一种活的微生物,当给予足够的量时,会给宿主带来健康益处”,从而调节肠道菌群平衡,发挥预防及治疗疾病的功效。因此,在动物生产上使用益生菌替代抗生素对维持动物健康具有重要意义。Zhang等^[4]研究报道,益生菌能够影响肉鸡肠道微生物组成、短链挥发性脂肪酸浓度和游离脂肪酸受体2/3(FFAR2/3)基因表达。Khattab等^[5]研究发现,在粗蛋白质水平为14%的饲粮中添加益生菌可提高白羽肉鸭的生长性能。Roselli等^[6]研究报道,益生菌具有调节猪肠道菌群、维持肠道健康和抗病的免疫调节作用。Ma等^[7]研究表明,在母猪饲粮中添加益生菌能够通过改善肠道菌群提高哺乳仔猪的免疫和抗氧化功能。Zhang等^[8]在饲粮中添加等量的丁酸梭菌(Clostridium butyricum)、粪肠球菌、枯草芽孢杆菌和植物乳杆菌(Lactiplantbacillus plantarum),结果表明饲粮中添加外源益生菌能够通过调节肠道菌群组成影响中国鲈鱼生长性能和肠道健康。Shehu等^[9]报道,酵母等益生菌及其提取物是天然抗氧化剂的优良来源,能够缓解热应激,促进肉兔生长。然而,近年来关于益生菌的研究主要集中在猪、禽和水产上,在肉兔生产上的研究报道相对较少。因此,本试验旨在研究饲粮中添加植物乳杆菌、戊糖片球菌(Pediococcus pentosaceus)和丁酸梭菌3种不同益生菌对肉兔生长性能、屠宰性能、回肠黏膜免疫指标和盲肠发酵指标的影响,为益生菌在肉兔生产中应用提供理论参考。

1 材料与方法

1.1 试验设计

试验选用28日龄断奶、体重相近、健康状况良好的商品伊拉肉兔400只(公母各占1/2),随机分为4组,每组10个重复,每个重复10只。对照组饲喂基础饲粮,试验组分别在基础饲粮中添加10⁸ CFU/kg植物乳杆菌、戊糖片球菌和丁酸梭菌。基础饲粮参考《肉兔营养需要量》(NY/T 4049—2021)^[10]行业标准配制而成,其组成及营养水平见表1。预试期7 d,正试期40 d。

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)%

原料 Ingredients	含量 Content	营养水平 Nutrient levels²⁾	含量 Content
玉米Corn	5.0	消化能DE/(MJ/kg)	10.23
豆粕Soybean meal	8.0	干物质DM	89.82
大麦Barley	6.0	粗蛋白质CP	16.12
麸皮Wheat bran	15.0	粗脂肪EE	2.80
玉米胚芽粕Corn germ meal	16.0	粗纤维CF	17.38
玉米皮Corn husk	17.0	中性洗涤纤维NDF	38.74
苜蓿草粉Alfalfa meal	15.0	酸性洗涤纤维ADF	23.08
豆秸粉Soybean straw powder	7.0	酸性洗涤木质素ADL	6.29
稻壳粉Rice hull powder	8.0	粗灰分Ash	9.03
磷酸氢钙CaHPO₄	1.5	钙Ca	0.95
氯化钠NaCl	0.5	总磷TP	0.45
预混料Premix¹⁾	1.0	赖氨酸Lys	0.60
合计Total	100.0	蛋氨酸+半胱氨酸Met+Cys	0.65

1)预混料为每千克饲粮提供 Premix provided the following per kg of the diet: VA 8 000 IU,VD₃ 1 500 IU,VE 45 mg,VK₃ 2.0 mg,VB₁ 1.0 mg,VB₂ 3.0 mg,VB₆ 1.5 mg,烟酸 nicotinic acid 30 mg,泛酸 pantothenic acid 50 mg,叶酸 folic acid 0.5 mg,氯化胆碱 choline chloride 100 mg,Fe 50 mg,Cu 10 mg,Zn 50 mg,Mn 10 mg,I 0.5 mg,Se 0.05 mg,赖氨酸 Lys 1.5 g,蛋氨酸 Met 0.5 g。

2)消化能为计算值,其余为实测值。DE was a calculated value, while the others were measured values.

1.2 样品采集与制备

试验结束前1天晚上禁食12 h,试验结束当天,每个重复选取1只与该重复平均体重相近的试验兔屠宰,统计屠宰性能;同时采集空肠样品,置于4%多聚甲醛固定,用于空肠组织形态结构测定;采集回肠和盲肠内容物样品,液氮冻存,分别用于回肠黏膜免疫指标和盲肠发酵指标测定。

1.3 测定指标和方法

1.3.1 生长性能

在试验正式开始(35日龄)和结束当日(75日龄),称量每个重复试验兔重量并计算平均日增重(ADG);统计试验期间每个重复试验兔总采食量,计算平均日采食量(ADFI)和料重比(F/G);统计试验期间每个重复试验兔发生腹泻的只数和成活的只数,计算腹泻率和成活率。计算公式如下:

平均日增重(g/d)=(末重-初重)/(试验天数×试验兔只数);

平均日采食量(g/d)=每个重复总采食量/(试验天数×试验兔只数);

料重比=平均日采食量/平均日增重;

腹泻率(%)=(发生腹泻的只数/10)×100;

成活率(%)=(成活的只数/10)×100。

1.3.2 屠宰性能

试验结束前1天晚上禁食12 h,试验结束当天每个重复选取1只与该重复平均体重相近的试验兔屠宰,称重并记录宰前活体重。颈部放血处死后,立即剥皮,在腕关节处去除前肢及在跗关节处去后肢,移走肠道及内容物和泌尿生殖器官后在第1颈椎处去头,同时去除气管、食管,并保留肝脏(摘去胆囊)、肾脏及肾周脂肪称重,即为半净膛胴体重。半净膛胴胴体重去心脏、肝脏、肾脏及肾周脂肪后称重即为全净膛胴体重,并分别除以宰前活体重,即半净膛屠宰率和全净膛屠宰率。

1.3.3 内脏器官发育指标

对心脏、肺脏、肝脏(摘去胆囊)、肾脏和脾脏分别称重,并分别除以宰前活体重,即脏器指数。

1.3.4 空肠组织形态结构

将空肠样品进行修剪、脱水、包埋、切片、染色、封片。使用正置白光拍照显微镜(Eclipse Ci-L,Nikon公司,日本)拍照,选取小肠组织的目的区域进行40倍成像,成像完成后使用Image-Pro Plus 6.0分析软件分别测量每张切片中5处完整肠绒毛高度、绒毛宽度、隐窝宽度、黏膜层厚度和肌层厚度,取平均值作为最终值,同时计算绒毛高度/隐窝深度。

1.3.5 回肠黏膜免疫指标

回肠分泌型免疫球蛋白A(secretory immunoglobulin A,sIgA)、肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)、干扰素-γ(interferon-γ,IFN-γ)和白细胞介素-6(interleukin-6, IL-6)含量分别使用江苏酶免实业有限公司酶联免疫试剂盒(货号分别为:MM-33548、MM-0240、MM-0290和MM-0302)测定。

1.3.6 盲肠发酵指标

采用Mettler MP120型酸碱度计测定盲肠内容物的pH。称取约0.2 g盲肠内容物样本,加入1.5 mL提取液,匀浆过夜浸提,超声提取60 min,离心取上清液,针头式过滤器过滤后在高效液相色谱仪(Rigol L3000)上测定,RIGOL C18反相色谱柱(250 mm×4.6 mm×5 μm),流动相的配制:800 mL水加1.56 g磷酸二氢钠混匀。用磷酸调节pH至4~5,最后加入16 mL甲醇。设置进样量10 μL,流速0.8 mL/min,柱温30 ℃,走样时间为30 min,紫外检测波长214 nm。利用铵态氮在强碱性介质中与次氯酸盐和苯酚作用,生成水溶性染料靛酚蓝,在625 nm处有特征吸收峰,吸光值与氨态氮含量成正比,进行氨态氮含量测定。

1.4 数据处理

采用SAS 9.1.3统计软件中的GLM程序进行数据的方差分析,并用Duncan氏法进行多重比较,结果以平均值和均方根误差(RMSE)表示,P<0.05为差异显著。

2 结果

2.1 饲粮中添加不同益生菌对肉兔生长性能的影响

由表2可知,饲粮中添加不同益生菌对肉兔末重、平均日增重、平均日采食量和料重比均无显著影响(P>0.05)。然而,饲粮中添加不同益生菌有降低肉兔腹泻率的趋势(P=0.059 5),有提高肉兔成活率的趋势(P=0.060 3)。

表2 饲粮中添加不同益生菌对肉兔生长性能的影响

Table 2 Effects of dietary adding different probiotics on growth performance of meat rabbits (n=10)

项目 Items	组别Groups				均方根误差 RMSE	P值 P-value
项目 Items	对照 Control	植物乳杆菌 Lactiplantbacillus plantarum	戊糖片球菌 Pediococcus pentosaceus	丁酸梭菌 Clostridium butyricum	均方根误差 RMSE	P值 P-value
初重IBW/g	961.7±16.7	939.0±17.2	958.0±11.3	936.0±15.3	48.413 7	0.557 6
末重FBW/g	2 632.2±26.5	2 582.6±19.6	2 597.3±16.2	2 563.0±34.9	80.153 6	0.280 9
平均日增重ADG/(g/d)	41.79±0.49	41.09±0.64	40.98±0.19	40.68±0.68	1.695 8	0.526 9
平均日采食量ADFI/(g/d)	171.17±2.43	174.94±2.99	177.03±3.14	178.59±3.74	9.836 2	0.376 2
料重比F/G	4.10±0.08	4.27±0.11	4.32±0.07	4.40±0.11	0.300 1	0.172 5
成活率Survival rate/%	89.00±2.77	92.00±3.27	91.00±3.14	92.00±2.49	8.279 6	0.060 3
腹泻率Diarrhea rate/%	11.00±3.79	8.00±3.27	9.00±2.77	7.00±3.00	9.206 2	0.059 5

	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 饲粮中添加不同益生菌对肉兔屠宰性能的影响

由表3可知,饲粮中添加不同益生菌对肉兔宰前活体重、半净膛胴体重、全净膛胴体重、半净膛屠宰率和全净膛屠宰率均无显著影响(P>0.05)。

表3 饲粮中添加不同益生菌对肉兔屠宰性能的影响

Table 3 Effects of dietary adding different probiotics on slaughter performance of meat rabbits (n=10)

项目 Items	组别Groups				均方根误差 RMSE	P值 P-value
项目 Items	对照 Control	植物乳杆菌 Lactiplantbacillus plantarum	戊糖片球菌 Pediococcus pentosaceus	丁酸梭菌 Clostridium butyricum	均方根误差 RMSE	P值 P-value
宰前活体重 Pre-slaughter body weight/g	2 607.0±65.6	2 550.0±65.1	2 605.6±50.8	2 633.0±80.8	210.094 1	0.841 4
半净膛胴体重 Semi clean carcass weight/g	1 366.7±32.8	1 326.9±38.3	1 362.0±35.3	1 385.9±50.4	125.747 8	0.766 5
全净膛胴体重 Full clean carcass weight/g	1 226.2±33.5	1 203.2±35.3	1 228.7±34.0	1 257.9±43.6	116.494 0	0.775 3
半净膛屠宰率 Semi clean slaughter ratio/%	52.45±0.33	52.03±0.65	52.23±0.62	52.57±0.49	1.697 2	0.899 7
全净膛屠宰率 Full clean slaughter ratio/%	47.03±0.46	47.18±0.63	47.12±0.69	47.73±0.45	1.792 6	0.817 7

2.3 饲粮中添加不同益生菌对肉兔内脏器官发育指标的影响

由表4可知,饲粮中添加不同益生菌对肉兔心脏指数、肺脏指数、肝脏指数和脾脏指数均无显著影响(P>0.05)。然而,丁酸梭菌组的肾脏指数较其他各组有降低的趋势(P=0.076 2)。

表4 饲粮中添加不同益生菌对肉兔内脏器官发育指标的影响

Table 4 Effects of dietary adding different probiotics on visceral organ development indexes of meat rabbits (n=10)g/kg

项目 Items	组别Groups				均方根误差 RMSE	P值 P-value
项目 Items	对照 Control	植物乳杆菌 Lactiplantbacillus plantarum	戊糖片球菌 Pediococcus pentosaceus	丁酸梭菌 Clostridium butyricum	均方根误差 RMSE	P值 P-value
心脏指数Heart index	3.14±0.14	3.03±0.14	3.07±0.12	3.14±0.13	0.423 2	0.910 4
肺脏指数Lung index	6.67±0.45	6.51±0.34	6.42±0.49	6.25±0.17	1.217 6	0.889 2
肝脏指数Liver index	34.19±1.90	32.77±0.99	33.23±1.39	30.63±1.59	4.752 8	0.403 2
肾脏指数Kidney index	7.43±0.17	7.13±0.31	7.21±0.26	6.55±0.16	0.748 9	0.076 2
脾脏指数Spleen index	0.95±0.09	0.78±0.07	0.84±0.05	0.76±0.03	0.204 4	0.168 9

2.4 饲粮中添加不同益生菌对肉兔空肠组织形态结构的影响

由表5可知,饲粮中添加不同益生菌对肉兔空肠绒毛高度和隐窝深度影响显著(P<0.05);其中,试验组的空肠绒毛高度显著高于对照组(P<0.05),丁酸梭菌组的空肠隐窝深度显著高于其他各组(P<0.05)。另外,戊糖片球菌组和丁酸梭菌组组的空肠黏膜层厚度有高于对照组和植物乳杆菌组的趋势(P=0.086 0),植物乳杆菌组和戊糖片球菌组的空肠绒毛高度/隐窝深度有高于对照组和丁酸梭菌组的趋势(P=0.051 6)。饲粮中添加不同益生菌对肉兔空肠绒毛宽度和肌层厚度无显著影响(P>0.05)。

表5 饲粮中添加不同益生菌对肉兔空肠组织形态结构的影响

Table 5 Effects of dietary add different probiotics on histological structure of jejunum of meat rabbits (n=10)

项目 Items	组别Groups				均方根误差 RMSE	P值 P-value
项目 Items	对照 Control	植物乳杆菌 Lactiplantbacillus plantarum	戊糖片球菌 Pediococcus pentosaceus	丁酸梭菌 Clostridium butyricum	均方根误差 RMSE	P值 P-value
绒毛高度Villus height/μm	686.06±41.41^b	831.45±28.86^a	830.28±37.35^a	825.96±45.42^a	122.530 5	0.027 5
隐窝深度Crypt depth/μm	110.29±5.93^b	114.10±4.98^b	114.69±7.18^b	138.68±5.50^a	18.825 6	0.006 8
绒毛宽度Villus width/μm	106.15±4.73	113.86±7.15	117.10±7.49	120.90±5.05	19.698 4	0.398 4
黏膜层厚度 Mucosal layer thickness/μm	930.80±28.21	984.00±43.24	1 019.20±47.59	1 081.23±42.12	129.458 0	0.086 0
肌层厚度 Muscle layer thickness/μm	127.44±10.77	141.20±11.76	157.75±7.84	130.70±8.91	31.433 3	0.149 5
绒毛高度/隐窝深度V/C	6.26±0.32	7.45±0.48	7.45±0.49	6.08±0.46	1.400 8	0.051 6

2.5 饲粮中添加不同益生菌对肉兔回肠黏膜免疫指标的影响

由表6可知,饲粮中添加不同益生菌对肉兔回肠sIgA、TNF-α和IFN-γ含量影响显著(P<0.05);其中,试验组的回肠sIgA含量显著高于对照组(P<0.05),回肠TNF-α和IFN-γ含量显著低于对照组(P<0.05)。饲粮中添加不同益生菌对肉兔回肠IL-6含量无显著影响(P>0.05)。

表6 饲粮中添加不同益生菌对肉兔回肠黏膜免疫指标的影响

Table 6 Effects of dietary adding different probiotics on ileum mucosal immune indexes of meat rabbits (n=10)

项目 Items	组别Groups				均方根误差 RMSE	P值 P-value
项目 Items	对照 Control	植物乳杆菌 Lactiplantbacillus plantarum	戊糖片球菌 Pediococcus pentosaceus	丁酸梭菌 Clostridium butyricum	均方根误差 RMSE	P值 P-value
分泌型免疫球蛋白A sIgA/(mg/g)	3.96±0.16^b	4.89±0.18^a	5.19±0.19^a	5.09±0.16^a	0.550 8	<0.000 1
肿瘤坏死因子-α TNF-α/(ng/g)	5.14±0.26^a	4.08±0.23^b	3.81±0.33^b	3.96±0.22^b	0.833 6	0.011 1
干扰素-γ IFN-γ/(ng/g)	7.00±0.37^a	5.60±0.44^b	5.41±0.29^b	4.97±0.30^b	1.121 3	0.009 3
白细胞介素-6 IL-6/(ng/g)	1.24±0.04	1.34±0.06	1.33±0.10	1.24±0.10	0.242 9	0.699 7

2.6 饲粮中添加不同益生菌对肉兔盲肠发酵指标的影响

由表7可知,饲粮中添加不同益生菌对肉兔盲肠内容物pH及氨态氮、乙酸、丁酸和总挥发性脂肪酸含量影响显著(P<0.05);其中,试验组的盲肠内容物pH显著低于对照组(P<0.05),试验组的盲肠内容物乙酸和总挥发性脂肪酸含量显著高于对照组(P<0.05),丁酸梭菌组的盲肠内容物氨态氮含量显著低于对照组(P<0.05)。饲粮中添加不同益生菌对肉兔盲肠内容物丙酸含量和乙酸/(丙酸+丁酸)无显著影响(P>0.05)。

表7 饲粮中添加不同益生菌对肉兔盲肠发酵指标的影响

Table 7 Effects of dietary adding different probiotics on cecal fermentation indexes of meat rabbits (n=10)

项目 Items	组别Groups				均方根误差 RMSE	P值 P-value
项目 Items	对照 Control	植物乳杆菌 Lactiplantbacillus plantarum	戊糖片球菌 Pediococcus pentosaceus	丁酸梭菌 Clostridium butyricum	均方根误差 RMSE	P值 P-value
pH	7.04±0.11^a	6.73±0.07^b	6.82±0.07^b	6.61±0.09^b	0.270 3	0.008 7
氨态氮 Ammoniacal nitrogen/(μg/g)	74.93±1.74^a	71.47±1.84^ab	73.29±1.35^ab	66.34±1.59^b	3.346 3	0.032 5
乙酸Acetic acid/(mg/g)	0.90±0.05^b	1.25±0.09^a	1.19±0.07^a	1.13±0.06^a	0.216 6	0.005 0
丙酸Propionic acid/(mg/g)	0.13±0.01	0.14±0.01	0.12±0.02	0.10±0.02	0.045 7	0.242 7
丁酸Butyric acid/(mg/g)	0.69±0.08^b	1.03±0.08^a	0.76±0.10^b	1.11±0.08^a	0.270 6	0.002 4
总挥发性脂肪酸 Total volatile fatty acids/(mg/g)	1.71±0.08^c	2.42±0.10^a	2.07±0.11^b	2.35±0.09^ab	0.306 7	<0.000 1
乙酸/(丙酸+丁酸) Acetic acid/(propionic acid+butyric acid)	1.21±0.14	1.14±0.14	1.55±0.23	0.98±0.10	0.499 8	0.089 0

3 讨论

益生菌是有益微生物经培养、发酵、干燥等特殊工艺制成的生物制剂或活菌制剂,肠道益生菌与动物机体生长发育、营养代谢和健康密切相关^[11]。益生菌种类繁多,目前用于饲用益生菌制剂的菌种主要有:乳酸菌、芽孢杆菌、酵母菌等^[12]。乳酸菌具有抗菌、调节免疫反应、维持肠上皮屏障和抗氧化等功能,在促进健康和预防疾病方面发挥着关键作用^[13]。植物乳杆菌和戊糖片球菌作为乳酸菌的成员,具有抗炎、抗氧化、调节动物肠道菌群环境、提高机体免疫能力、改善生长性能等功能^[14]。Funk等^[15]以每天0.8 mL/kg BW的剂量将植物乳杆菌制剂添加到妊娠期山羊的饲粮中,结果表明植物乳杆菌制剂对羔羊生长发育具有促进作用。Sagada等^[16]研究报道,饲粮中添加植物乳杆菌能够提高幼年黑海鲷鱼生长性能、抗氧化能力和免疫功能。Hashemi等^[17]研究表明,使用戊聚糖片球菌作为益生菌能够提高凡纳滨对虾的成活率。丁酸梭菌是一种以产丁酸而得名的厌氧菌,其特有的代谢产物决定了其广泛的生物学功能。Cai等^[18]研究报道,饲粮中添加有效剂量(0.05%)的丁酸梭菌可以提高热应激状态下山羊的生长性能。饲粮中添加丁酸梭菌可以改善环江小型猪胴体性状和肉品质^[19]。另外,饲粮中添加丁酸梭菌可改善黄羽肉鸡母鸡的繁殖参数、蛋品质、孵化性能和后代的生长性能^[20]。本研究发现,饲粮中添加植物乳杆菌、戊糖片球菌和丁酸梭菌有降低试验期肉兔腹泻率的趋势,有提高成活率的趋势,并未观察到明显的生长性能和屠宰性能的改变,这可能是由于益生菌的效果在不同物种之间存在差异所致。

大量研究表明,益生菌能够改善肠道形态结构,维持肠道健康,提高机体免疫力。An等^[21]研究报道,饲粮中添加400 mg/kg植物乳杆菌可改善肉鸭的肠道健康,提高回肠黏膜sIgA含量。Dong等^[22]研究报道,戊糖片球菌对结肠炎具有保护作用,并能调节小鼠肠道菌群和免疫应答。Wang等^[23]研究发现,戊糖片球菌能够提高小鼠肠道黏膜屏障,通过降低炎症反应来降低副溶血性弧菌感染。此外,戊糖片球菌在敌百虫引起的动物肠道损伤中具有缓解氧化应激、恢复肠道菌群的功能^[24]。丁酸梭菌作为益生菌具有抑制肠道致病菌侵入的效果^[25]。饲粮中添加丁酸梭菌能够调节肉鸡肠道黏膜上皮细胞,促进免疫和肠道屏障功能,改善肉鸡生长性能^[26-27]。Yu等^[28]研究报道,饲粮中添加丁酸梭菌能够通过改善肠道结构和功能促进鹅的生长。Huang等^[29]研究发现,饲粮中添加丁酸梭菌的肉兔回肠绒毛高度和绒毛高度/隐窝深度显著升高,隐窝深度降低;此外,饲粮中添加丁酸梭菌显著提高了肉兔回肠sIgA的分泌。本研究发现,饲粮中添加益生菌的试验组回肠绒毛高度显著提高,丁酸梭菌组的隐窝深度显著高于其他各组。此外,饲粮中添加益生菌能够提高回肠sIgA含量,降低炎症因子TNF-α和IFN-γ的含量。

肉兔盲肠占肠道总容积的40%,盲肠内富含大量微生物,与宿主保持相对平衡和稳定的状态,在营养物质消化、吸收、代谢和肠道黏膜免疫调节中发挥重要作用。肉兔盲肠微生物可以发酵纤维素产生挥发性脂肪酸为肉兔提供能量^[30]。Liu等^[31]研究报道,丁酸梭菌能够通过增加肠道菌群多样性,提高挥发性脂肪酸含量,增强肠道屏障功能和免疫系统,改善北京鸭肠道健康。Li等^[32]研究表明,饲喂丁酸乳杆菌能够改变荷斯坦母牛瘤胃挥发性脂肪酸含量及组成。饲粮中添加丁酸梭菌能够改善断奶仔猪的肠道健康,改变断奶仔猪肠道挥发性脂肪酸等代谢产物的含量,能够为肠道健康提供潜在的益处^[33-34]。本研究发现,饲粮中添加益生菌后盲肠内容物中乙酸和总挥发性脂肪酸含量升高,pH降低,且丁酸梭菌组盲肠内容物氨态氮含量显著降低,这与前人报道基本一致。

4 结论

饲粮中添加益生菌并不能改变商品肉兔生长性能和屠宰性能,但能够提高机体肠道黏膜免疫性能,改变盲肠内容物挥发性脂肪酸组成,对维持肉兔肠道健康具有一定促进作用。综合本试验结果,丁酸梭菌对维持肉兔肠道形态结构效果较好。

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