益生菌对载鸭水体和产蛋性能的影响及其分子机制

doi:10.3864/j.issn.0578-1752.2015.11.017

摘要/Abstract

摘要： 【目的】研究饲粮添加芽孢杆菌和载鸭水体应用光合细菌对载鸭水体和鸭产蛋性能的影响及其分子机制。【方法】采用“地面平养＋水池”养殖模式，以2×2因子试验设计，选择同批次连城白鸭和樱桃谷鸭杂交后代蛋鸭240只，随机分为4组，每组6个重复，每个重复10只，每个重复独立一栏。两处理因子分别为饲粮添加有效活菌含量为8×10⁷ cfu/kg的枯草芽孢杆菌和载鸭水体应用光合细菌至水体有效活菌含量为1.8×10¹² cfu/m³。对照组为不应用芽孢杆菌和光合细菌，光合组为只应用光合细菌，芽孢组为只添加芽孢杆菌，联合组为既添加芽孢杆菌又应用光合细菌。预饲7 d，试验期120 d。试验期间统计每10 d的平均产蛋率、平均采食量和平均料蛋比；每10 d采集水样用于检测大肠杆菌和沙门+志贺氏菌数量；每15 d采集水样和血样用于检测内毒素浓度。试验结束时，采集新鲜粪样用于检测大肠杆菌和沙门+志贺氏菌数量；取十二指肠、空肠和回肠黏膜用于检测内毒素水平；取空肠、回肠和脾脏组织用于炎症和凋亡相关基因表达。试验鸭自由饮用池水、自由采食、自然光照。水体和血液内毒素、采食量、产蛋率、料蛋比、水体大肠杆菌和沙门+志贺氏菌数据采用重复测量的线性混合模型分析，LSD修正法多重比较。粪便微生物数量、肠道黏膜内毒素水平和组织基因表达数据采用单因子方差分析，Duncan法多重比较。【结果】芽孢杆菌和光合细菌不显著影响鸭采食量（P＞0.05）、空肠Caspase-3和Bcl-2 mRNA表达（P＞0.05）、回肠Bcl-2 mRNA表达（P＞0.05）、以及脾脏IL-1β mRNA表达（P＞0.05），但芽孢杆菌和光合细菌显著提高蛋鸭产蛋率（P ＜0.001），显著降低蛋鸭料蛋比（P＜0.001）、载鸭水体（P＜0.001）、十二指肠（P＜0.01）和回肠（P＜0.01）内毒素浓度、粪便大肠杆菌数（P＜0.001）、空肠TLR4（P＜0.05）和回肠Caspase-3（P＜0.05）mRNA表达水平。采食量、产蛋率、料蛋比、水体大肠杆菌、沙门+志贺氏菌和内毒素水平存在显著的时间效应（P＜0.001）。与芽孢组和联合组相比，对照组和光合组血液内毒素和粪便沙门+志贺氏菌数量显著提高，对照组空肠粘膜内毒素水平、载鸭水体沙门+志贺氏菌数量以及脾脏TNF-α mRNA表达水平显著提高。联合组水体大肠杆菌数量显著低于对照组、光合组和芽孢组。仅联合组回肠TLR4 mRNA表达水平显著低于对照组，脾脏IL-10 mRNA表达水平显著高于对照组。【结论】芽孢杆菌和光合细菌可能通过降低载鸭水体革兰氏阴性菌数量和内毒素水平、降低肠道和血液内毒素浓度、降低炎症反应和肠道细胞凋亡，提高蛋鸭产蛋性能。

关键词: 蛋鸭, 产蛋性能, 光合细菌, 芽孢杆菌, 内毒素

Abstract: 【Objective】 This study aims to investigate the effects of supplementation of Bacillus subtilis spores in the feed and addition of photosynthetic bacteria (PSB) on living water and laying performance of laying ducks and their mechanisms.【Method】A total of 240 laying ducks were randomly and equally allocated into 24 “slate floor bed + water pool” individual pens using a 2×2 factorial design with 6 replications per treatment. After 7-d pre-feeding, the ducks were received 2 main treatments: supplementation of Bacillus subtilis (viable count 8 × 10⁷ cfu/kg) in the feed, and addition of PSB (viable count 1.8 × 10¹² cfu/kg) to the living water. Neither supplementation of B. subtilis nor addition of PSB into living water was used as the control group (C group), supplementation of B. subtilis was used as the B. subtilis group (B group), addition of PSB into living water was used as the PSB group (P group), and both supplementation of B. subtilis and addition of PSB into living water was used as the combined group (BP group). In the 120-d experimental period, the average laying rate, average feed intake and average feed to egg ratio were calculated every10 days. The living water samples were collected every 10 days for measuring counts of E. coli and Salmonella + Shigella. The living water and blood samples were collected every 15 days for measuring LPS concentration. At the end of the experiment, the fresh feces were collected for measuring counts of E. coli and Salmonella + Shigella. The mucosa and tissues of duodenum, jejunum and ileum were collected for measuring LPS concentrations and mRNA expression of inflammation/apoptosis - related genes, respectively. During the experiment, the animals were maintained under natural photoperiod. Living water and feed were available to the ducks ad libitum. The linear mixed model of SPSS, for the analysis of repeated measurements, was used with comparisons using the LSD correction. The data with repeated measurements included LPS concentrations in living water and plasma, feed intake, laying rate, feed to egg ratio, and counts of E. coli and Salmonella + Shigella in living water. The one-way ANOVA was used for data including counts of E. coli and Salmonella + Shigella in feces, LPS concentrations of intestinal mucosa, and tissue gene expression. Post-hoc differences between treatment groups were further examined using Duncan test.【Result】Both Bacillus subtilis and PSB treatments did not significantly affect feed intake (P＞0.05), Caspase-3 and Bcl-2 mRNA expression in jejunum (P＞0.05), Bcl-2 mRNA expression in ileum (P＞0.05), and IL-1β mRNA expression in spleen, however, they significantly increased laying rate (P＜0.001), and decreased feed to egg ratio (P＜0.001), LPS concentrations of living water (P＜0.001), duodenum (P＜0.01) and ileum (P＜0.01), counts of E. coli in feces (P＜0.001), and mRNA expression of TLR4 in jejunum (P＜0.05) and Caspase-3 in ileum (P＜0.05). There was significant effect of time on feed intake, laying rate, feed to egg ratio, LPS concentration and counts of E. coli and Salmonella + Shigella in living water (P＜0.01). Compared with B and BP groups, LPS concentration in plasma and counts of Salmonella + Shigella in feces in C and P groups were increased (P＜0.05), and LPS concentration in jejunum, counts of Salmonella + Shigella in living water and TNF-α mRNA expression in spleen were also increased (P＜0.05). The count of E. coli in living water in BP group was lower than in C, B and P groups (P＜0.05). Only in BP group was TLR4 mRNA expression in ileum lower (P＜0.05), and IL-10 mRNA expression higher than in C group (P＜0.05).【Conclusion】The results indicate that B. subtilis and PSB treatments improved laying performance of laying ducks by decreasing the counts of Gram-negative bacteria and level of LPS in cultivated water, and decreasing the level of LPS in intestinal mucosa and plasma and inflammatory response.

Key words: laying ducks, laying performance, photosynthetic, bacillus, lipopolysaccharide

应诗家，许爱红，戴子淳，郗正林，何宗亮，匡伟，赵伟，吴云良，施振旦. 益生菌对载鸭水体和产蛋性能的影响及其分子机制[J]. 中国农业科学, 2015, 48(11): 2262-2269.

YING Shi-jia, XU Ai-hong, DAI Zi-chun, XI Zheng-lin, HE Zong-liang, KUANG Wei, ZHAO Wei, WU Yun-liang, SHI Zhen-dan . Effects of Pro-Biotic Microbes on Living Water and Laying Performance in Layer Ducks and Molecular Mechanisms[J]. Scientia Agricultura Sinica, 2015, 48(11): 2262-2269.

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