动物营养学报    2021, Vol. 33 Issue (8): 4332-4341    PDF    
枯草芽孢杆菌对断奶仔猪生长性能影响的Meta分析
杨宽民1 , 申远航2 , 王丽1 , 蒋宗勇1     
1. 广东省农业科学院动物科学研究所, 农业农村部华南动物营养与饲料重点实验室, 畜禽育种国家重点实验室, 岭南现代农业科学与技术广东省实验室茂名分中心, 广东省畜禽育种与营养研究重点实验室, 广州 510640;
2. 深圳比利美英伟营养饲料有限公司, 深圳 518103
摘要: 为探究枯草芽孢杆菌对断奶仔猪生长性能的影响,本试验进行了系统回顾和Meta分析。计算机检索中国知网、维普、万方、Science Direct、Web of Science数据库,查找枯草芽孢杆菌对断奶仔猪生长性能影响的随机对照试验。检索年限从建库到2020年,按照纳入和排除标准筛选文献、提取资料,用Review Manager 5.3软件进行Meta分析,并对纳入研究进行发表偏倚分析、亚组分析和敏感性分析。纳入18个研究,共计样本1 598例。Meta分析结果显示:枯草芽孢杆菌组平均日增重显著高于空白组[P < 0.000 01,标准化均数差(SMD)=1.34,95%置信区间(95% CI):0.76~1.93];料重比显著低于空白组(P=0.000 3,SMD=-0.86,95% CI:-1.33~-0.40);腹泻率显著低于空白组(P=0.000 2,SMD=-3.07,95% CI:-4.68~-1.46),但平均日采食量与空白组不存在统计学差异(P=0.12,SMD=0.33,95% CI:-0.08~0.74)。综上所述,枯草芽孢杆菌可提高断奶仔猪平均日增重,降低料重比、腹泻率,对平均日采食量影响不大,以上结果可为断奶仔猪生产上添加剂的选择提供理论依据。
关键词: 枯草芽孢杆菌    断奶仔猪    生长性能    Meta分析    
Effects of Bacillus subtilis on Growth Performance of Weaned Pglets: a Meta-Analysis
YANG Kuanmin1 , SHEN Yuanhang2 , WANG Li1 , JIANG Zongyong1     
1. Guangdong Key Laboratory of Animal Breeding and Nutrition, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming Branch, State Key Laboratory of Livestock and Poultry Breeding, Key Laboratory of Animal Nutrition and Feed Science in South China Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China;
2. Shenzhen Premix Nutrition Co., Ltd., Shenzhen 518103, China
Abstract: The aim of this systematic review and Meta-analysis was to explore the effects of Bacillus subtilis on the growth performance of weaned piglets. The study conducted a literature search about randomized controlled trials on the effects of Bacillus subtilis on the growth performance of weaned piglets in English and Chinese databases, including CNKI, CQVIP, Wan Fang Data, Science Direct and Web of science databases. The retrieval period was from the establishment of the database to 2020. According to the inclusion and exclusion criteria, we formulated, the literatures were screened and the data were extracted. The Meta-analysis was conducted using Review Manager V5.3 software. Publication bias analysis, subgroup analysis and sensitivity analysis were also performed. A total of 1 742 samples were included in 18 studies. The results of Meta-analysis showed that average daily gain of Bacillus subtilis group was significantly higher than that of control group[P < 0.000 01, standardized mean difference (SMD)=1.34, 95% confidence interval (CI): 0.76 to 1.93]. Meanwhile, feed to gain ratio of Bacillus subtilis group was significantly lower than that of control group (P=0.000 3, SMD=-0.86, 95% CI: -1.33 to -0.40). Moreover, lower diarrhea rate was found in Bacillus subtilis group than control group (P=0.000 2, SMD=-3.07, 95% CI: -4.68 to -1.46), but there was no significant difference in feed intake between the two groups (P=0.12, SMD=0.33, 95% CI: -0.08 to 0.74). This study demonstrates that Bacillus subtilis improves the average daily gain of weaned piglets, reduces the feed to gain ratio and diarrhea rate, and doesn't influence the average daily feed intake. The above results provide a theoretical basis for the selection of additives in the production of weaned piglets.
Key words: Bacillus subtilis    weaned piglets    growth performance    Meta-analysis    

随着一系列禁抗措施的颁布和实施,我国畜牧业已进入全面禁抗时代。断奶仔猪阶段是猪场生产中重要的生产阶段,仔猪体内没有产生足够的抗体,肠道微生物生态平衡不健全,胃酸和消化酶分泌量较低[1-2],断奶后由于饲粮、环境、饲养制度等的转变,对仔猪造成极大的应激。断奶应激是影响养猪生产效率的重要因素,解决此问题也成为养猪业在禁抗背景下的一大难题。益生菌具有性质较稳定、无毒、无不良反应、不产生耐药性、无残留、无污染等优点[3-5]。枯草芽孢杆菌是常用的益生菌菌种,具有耐高温、制粒稳定、过瘤胃稳定等特点,通过嗜氧、厌氧繁殖可较好地抑制有害菌的生长,具有较强的酶活性,可促进营养物质消化吸收[6-8]。在菌株代谢过程中可产生短杆菌肽等活性物质,促进肠道形成厌氧环境,可提高仔猪平均日增重、平均日采食量、降低腹泻率等,具有缓解断奶应激的应用价值[9]。然而,检索发现枯草芽孢杆菌在断奶仔猪上应用的研究结果并不一致,需要进一步系统循证性研究与评价。

Meta分析是运用统计学方法对多个研究结果进行系统、定量和客观的综合分析,对效应指标进行准确和科学的评估,以达到增大样本数量、提高检验效能的效果,为临床决策提供有力的依据[10-11]。本研究采用Meta分析方法,以枯草芽孢杆菌为干预措施,以断奶仔猪的平均日增重、平均日采食量、料重比和腹泻率为结局指标,对发表的相关历史研究进行系统评价,为枯草芽孢杆菌在断奶仔猪生产上的应用提供参考。

1 材料与方法 1.1 检索策略

计算机检索中国知网、万方、维普、Science Direct、Web of Science数据库。检索年限从建库至今。中文检索关键词包括:断奶仔猪、仔猪、枯草芽孢杆菌、益生菌,英文检索关键词包括:weaned piglets、piglets、Bacillus subtilis、probiotics,根据不同数据库的特征分别进行主题词联合自由词相结合的方式收集文献。

1.2 文献的纳入和排除标准

研究类型:已发表的随机对照试验;研究对象:断奶仔猪,必须是健康无疾病;干预措施:饲粮中添加益生菌并以空白作为对照;结局指标:平均日增重、平均日采食量、料重比、腹泻率;纳入文献中给出了指标的平均值和标准差(standard deviation, SD)或标准误(standard error of mean, SE)。

有以下情况应排除:1)重复发表、质量较低的文献;2)综述类文章、无空白对照组的文献;3)数据不全或数据明显有误的文献;4)试验组干预措施为益生菌以外的治疗方法或枯草芽孢杆菌与化学药物联合用药的情况。

1.3 数据资料提取

仔细阅读全文后,由2名评价者独立提取评价材料。从文献中提取以下数据:1)题目、第一作者、发表年份;2)研究对象、试验组动物总数、对照组动物总数、给药途径、饲养天数、动物品种、初重、末重和枯草芽孢杆菌添加量,如文中有多个添加量只提取中等添加量,排除低剂量和高剂量;3)平均日增重、平均日采食量、料重比、腹泻率。各研究中如有缺乏的资料尝试通过电话或邮件与作者进行联系补充。

1.4 统计方法

运用Review Manager 5.3软件进行数据分析并绘制相关图表。由于所纳入文献结局指标为连续型变量,且研究间测量方法、时间及表达单位不完全一致,故选择标准化均数差(standardized mean difference, SMD)为效应尺度。考虑到多个研究之间潜在的临床或方法学异质性,使用了随机效应模型(randomized effect model, REM),该模型会对合并效应进行更保守的估计。如果未提供均值和标准差,将根据标准误差、95%可信区间(confidence interval, CI)或其他统计指标进行计算。为了评估异质性,使用了异质性检验I2统计量和卡方检验。概率值P < 0.05被认为是显著的。并对分析结果进行敏感性分析和亚组分析,评价结果的稳定性,探究异质性来源,以漏斗图检验其发表偏移。

2 结果与分析 2.1 文献检索与筛选结果

按照检索策略共得到文献636篇,阅读文献题目及摘要,排除非临床研究,综述、重复发表的文献578篇,阅读全文,排除为研究益生菌对猪某些性能的影响而进行分离和选择具有潜在益生菌活性的菌株的任何体内试验,排除对照组不是空白组的文献,共排除不符合纳入标准的文献39篇,最终纳入文献18篇。文献筛选流程如图 1所示。

图 1 文献筛选流程 Fig. 1 Literature screening process
2.2 纳入文献的基本特征

研究对象为断奶仔猪,试验组共799头,对照组799头。试验组均为枯草芽孢杆菌添加至饲粮中,对照组为空白对照组,基本特征见表 1

表 1 基本特征 Table 1 Basic characteristics
2.3 枯草芽孢杆菌对断奶仔猪平均日增重影响的森林图、敏感性分析、亚组分析和漏斗图

18项研究报告了枯草芽孢杆菌对断奶仔猪平均日增重的影响,分析中包括1 598头断奶仔猪,各研究间异质性较高(I2=96%>50%),采用随机效应模型进行分析,分析结果发现枯草芽孢杆菌组平均日增重显著高于空白组(P < 0.000 01, SMD=1.34, 95% CI:0.76~1.93,图 2)。这提示枯草芽孢杆菌可提高断奶仔猪的平均日增重。依次去除该分析中任一项文献后进行敏感性分析,结果发现整体异质性未发生明显变化,所以本项结局指标的分析结果较为稳健。按文献为中文和英文文献、2015年之前的文献和之后的文献进行亚组分析(表 2)。结果显示英文文献的异质性较小,试验组和空白组差异不显著,但可信区间有趋向右侧倾向,可能的原因为英文文献纳入数量少,其他结果与森林图结果一致。同时制作漏斗图进行发表偏移检测,发现对称性相对较好,说明其发表偏倚程度较小(图 3)。

Study or Subgroup:研究或亚组;Mean:均值;SD:标准差standard deviation;Total:样本数sample number;Weight:权重;Std.Mean Difference:标准化均数差;Random:随机效应模型random effects model;95% CI:95%置信区间95% confidence interval;Total (95% CI):总体95%置信区间total 95% confidence interval;Heterogeneity:异质性检验;Test for overall effect:总效应量检验。图 4图 6图 8同。The same as Fig. 4Fig. 6 and Fig. 8. 图 2 枯草芽孢杆菌对断奶仔猪平均日增重影响的森林图 Fig. 2 Forest map of effects of Bacillus subtilis on average daily gain of weaned piglets
表 2 结局指标亚组分析结果 Table 2 Results of subgroup analysis of outcome indicators
图 3 枯草芽孢杆菌对断奶仔猪平均日增重影响的漏斗图 Fig. 3 Funnel plot of effects of Bacillus subtilis on average daily gain of weaned piglets
2.4 枯草芽孢杆菌对断奶仔猪平均日采食量影响的森林图、敏感性分析、亚组分析和漏斗图

13项研究报告了枯草芽孢杆菌对断奶仔猪平均日采食量的影响,分析中包括1 178头断奶仔猪,各研究间异质性较高(I2=91%>50%),采用随机效应模型进行分析,分析结果发现枯草芽孢杆菌组采食量与空白组不存在统计学差异(P=0.12, SMD=0.33, 95% CI:-0.08~0.74,图 4)。这说明枯草芽孢杆菌对断奶仔猪平均日采食量无显著影响。依次去除该分析中任一项研究后进行敏感性分析,结果发现整体异质性未发生变化,所以本项结局指标的分析结果较为稳健。按文献为中文和英文文献、2015年之前文献和之后的文献进行亚组分析(表 2)。结果显示英文文献的异质性较小,试验组和空白组差异不显著,与森林图结果一致。而中文文献、2015年之前的文献和之后的文献都显示试验组平均采食量显著高于空白组,说明英文文献是异质性来源,但英文文献数量较少,后期应纳入更多的文献进行分析。同时制作漏斗图进行发表偏移检测,发现对称性相对较好,说明其发表偏倚程度较小(图 5)。

图 4 枯草芽孢杆菌对断奶仔猪平均日采食量影响的森林图 Fig. 4 Forest map of effects of Bacillus subtilis on average daily feed intake of weaned piglets
图 5 枯草芽孢杆菌对断奶仔猪平均日采食量影响的漏斗图 Fig. 5 Funnel plot of effects of Bacillus subtilis on average daily feed intake of weaned piglets
2.5 枯草芽孢杆菌对断奶仔猪料重比影响的森林图、敏感性分析、亚组分析和漏斗图

14项研究报告了枯草芽孢杆菌对断奶仔猪料重比的影响,分析中包括1 238头断奶仔猪,各研究间异质性较高(I2=93%>50%),采用随机效应模型进行分析,分析结果发现枯草芽孢杆菌组料重比极显著低于空白组(P=0.000 3, SMD=-0.86, 95% CI:-1.33~-0.40,图 6)。这提示枯草芽孢杆菌可显著降低断奶仔猪料重比。依次去除该分析中任一项研究后进行敏感性分析,结果发现整体异质性未发生明显变化,所以本项结局指标的分析结果较为稳健。按文献为中文和英文文献、2015年之前文献和之后文献进行亚组分析(表 2)。结果显示英文文献的异质性较小,试验组和空白组差异不显著,但可信区间有趋向右侧倾向,可能的原因为英文文献纳入数量少,其他结果与森林图结果一致。同时制作漏斗图进行发表偏移检测,发现对称性相对较好,说明其发表偏倚程度较小(图 7)。

图 6 枯草芽孢杆菌对断奶仔猪料重比影响的森林图 Fig. 6 Forest map of effects of Bacillus subtilis on feed to gain ratio of weaned piglets
图 7 枯草芽孢杆菌对断奶仔猪料重比影响的漏斗图 Fig. 7 Funnel plot of effects of Bacillus subtilis on feed to gain ratio of weaned piglets
2.6 枯草芽孢杆菌对断奶仔猪腹泻率影响的森林图、漏斗图和敏感性分析

5项研究报告了枯草芽孢杆菌对断奶仔猪腹泻率的影响,分析中包括406头断奶仔猪,各研究间异质性较高(I2=97%>50%),采用随机效应模型进行分析,分析结果发现枯草芽孢杆菌组腹泻率极显著低于空白组(P=0.000 2, SMD=-3.07, 95% CI:-4.68~-1.46,图 8)。这表明枯草芽孢杆菌具有明显降低断奶仔猪腹泻率的效果。依次去除该分析中任一项研究后进行敏感性分析,结果发现整体异质性未发生明显变化,所以本项结局指标的分析结果较为稳健。同时制作漏斗图进行发表偏移检测,发现文献小部分对称性相对较差,说明有一定的发表偏倚,但偏倚程度不大(图 9)。

图 8 枯草芽孢杆菌对断奶仔猪腹泻率影响的森林图 Fig. 8 Forest map of effects of Bacillus subtilis on diarrhea rate of weaned piglets
图 9 枯草芽孢杆菌对断奶仔猪腹泻率影响的漏斗图 Fig. 9 Funnel plot of effects of Bacillus subtilis on diarrhea rate of weaned piglets
3 讨论

众多学者发现,在饲粮中添加益生菌可改善动物的生长性能,可起到一定程度的饲用抗生素作用,如乳酸菌、酵母菌和枯草芽孢杆菌等。但实际生产中枯草芽孢杆菌的使用效果并不一致[29]。为此本文搜集大量同类研究利用Meta分析方法进行科学评价,以期为枯草芽孢杆菌在断奶仔猪上的应用提供有价值的参考。因Meta分析是建立在已有文献数据的基础之上,故结论可靠性和文献偏倚性受纳入文献质量的影响较大[30]。本研究也可能存在一定的局限性:1)各研究试验饲养环境、饲粮情况存在差异;2)所选择枯草芽孢杆菌的菌株和剂量可能不一致;3)各研究间品种、断奶策略的不同,可能对分析结果造成一定的影响。本结果也需要开展更多设计合理的大样本且试验周期足够长的随机对照试验,将进一步验证其效果。

枯草芽孢杆菌能够有效解决断奶仔猪腹泻问题,改善其生长性能[31]。研究表明,枯草芽孢杆菌对仔猪肠道形态和微生态菌群有益,可增加小肠绒毛高度、降低隐窝深度和增加绒隐比,从而增加小肠与营养物质的接触面积[6, 32],即可增强动物肠道对营养物质的消化吸收能力,提高平均日增重和料重比。饲粮中添加一定量的枯草芽孢杆菌还可增加乳酸杆菌等有益菌的数量,形成厌氧环境,抑制肠道内大肠杆菌等有害菌的增殖,改善仔猪肠道菌群平衡,保证动物的健康和促进生长。另外,有益菌数量增加可调节肠道pH,刺激免疫系统提高干扰素和巨噬细胞活性,提高机体免疫力[33],与降低腹泻率有关,与本研究结果相一致。因此,在仔猪饲粮中添加枯草芽孢杆菌可显著提高平均日增重,降低料重比和腹泻率。

4 结论

本文对纳入的18项研究(1 598头断奶仔猪的试验数据)进行Meta分析,分析结果稳定可靠,结果显示枯草芽孢杆菌可提高断奶仔猪平均日增重,降低料重比、腹泻率,对平均日采食量影响不大,表明枯草芽孢杆菌在断奶仔猪生产上具有较好的应用效果。

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