黄曲霉毒素B1、赭曲霉毒素A和玉米赤霉烯酮联合对奶山羊肠道微生物的影响

doi:10.16431/j.cnki.1671-7236.2017.12.004

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (12): 3418-3425.doi: 10.16431/j.cnki.1671-7236.2017.12.004

黄曲霉毒素B₁、赭曲霉毒素A和玉米赤霉烯酮联合对奶山羊肠道微生物的影响

范彩云¹, 苏娣¹, 李晓娇¹, 黄帅¹, 张养东², 郑楠², 刘光磊³, 刘祯⁴, 程建波¹

1. 安徽农业大学动物科技学院, 合肥 230036;
2. 中国农业科学院北京畜牧兽医研究所, 农业部奶产品质量安全风险评估实验室(北京), 北京 100193;
3. 上海光明荷斯坦牧业有限公司, 上海 200443;
4. 青岛市畜牧兽医研究所, 青岛 266100

修回日期:2017-10-26 出版日期:2017-12-20 发布日期:2017-12-20
通讯作者: 程建波 E-mail:chengjianboahau@163.com
作者简介:范彩云(1980-),女,内蒙古包头人,博士,研究方向:动物分子营养,E-mail:fancaiyunnmgbt@163.com
基金资助:
公益性行业（农业）科研专项（201303144）；长三角联合攻关项目（16395810103）；农业部948项目（2016-X37）

Effects of Aflatoxin B₁ Individual,Binary or Tertiary Mixed with Ochratoxin A and Zearalenone on Intestinal Microorganism of Dairy Goat

FAN Cai-yun¹, SU Di¹, LI Xiao-jiao¹, HUANG Shuai¹, ZHANG Yang-dong², ZHENG Nan², LIU Guang-lei³, LIU Zhen⁴, CHENG Jian-bo¹

1. School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
2. Ministry of Agriculture-Milk Risk Assessment Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
3. Shanghai Bright Holstan Co., Ltd., Shanghai 200443, China;
4. Qingdao Research Institute of Husbandry and Veterinary, Qingdao 266100, China

Revised:2017-10-26 Online:2017-12-20 Published:2017-12-20

摘要/Abstract

摘要：

为研究黄曲霉毒素B₁（AFB₁）、赭曲霉毒素A（OTA）和玉米赤霉烯酮（ZEN）联合对奶山羊肠道微生物组成及多样性的影响，本研究选取体况相近的健康崂山奶山羊20头，分为5组，每组4头，A组为对照组，饲喂不添加毒素的基础日粮；B、C、D、E组分别在日粮中添加50 μg/kg AFB₁+500 μg/kg ZEN、50 μg/kg AFB₁+100 μg/kg OTA、50 μg/kg AFB₁及50 μg/kg AFB₁+100 μg/kg OTA+500 μg/kg ZEN。预试期7 d，正试期14 d。在试验期第11天采集粪便样品，利用基因组DNA提取试剂盒提取粪便总DNA，对总DNA进行16S rRNA扩增，构建奶山羊肠道微生物16S rRNA基因克隆文库并进行数据分析。经稀释曲线和多样性指数分析显示，单一AFB₁或与其他毒素联合处理后奶山羊肠道微生物多样性有降低趋势，但差异不显著（P>0.05），而奶山羊肠道菌群总数在AFB₁与其他毒素的联合处理后显著降低（P<0.05）。对获得的OTUs代表序列进行物种注释后发现，厚壁菌门（Firmicutes）、拟杆菌门（Bacteroidetes）、软壁菌门（Tenericutes）为山羊肠道中主要的细菌类群；经LEfSe分析显示，AFB₁、OTA和ZEN 3种毒素的联合作用可显著增加琥珀酸弧菌科（Succinivibrionaceae）、考拉杆菌属（Phascolarctobacterium）和气单胞菌科（Aeromonadales）丰度。而未处理组的霍尔德曼氏菌（Holdemania）和BF311拟杆菌的丰度显著高于其他组（P<0.05）。以上结果表明，AFB₁、ZEN和OTA联合能影响山羊肠道微生物区系结构，且影响程度依次为：AFB₁+OTA+ZEN > AFB₁+OTA > AFB₁+ZEN > AFB₁。

关键词: 奶山羊; 霉菌毒素; 肠道微生物; 16S rRNA

Abstract:

This experiment was conducted to investigate the effects of aflatoxin B₁ (AFB₁) individual,binary or tertiary mixed with ochratoxin A (OTA) and zearalenone (ZEN) on the bacterial composition and diversity of dairy goat intestinal tract. Twenty lactating Laoshan dairy goats with similar conditions were randomly assigned to five treatments (n=4). The dairy goats in control group (group A) were fed basal diet,while that of groups B,C,D and E were fed basal diet supplementation 50 μg/kg AFB₁+500 μg/kg ZEN,50 μg/kg AFB₁+100 μg/kg OTA,50 μg/kg AFB₁, 50 μg/kg AFB₁+100 μg/kg OTA+500 μg/kg ZEN,respectively. The experimental period consisted of 7 days for adaptation and 14 days for feeding mycotoxins. Faeces samples were collected on the 11th of training. The total DNA of feces was extracted using genomic DNA extraction kit,then 16S rRNA was amplified,and the 16S rRNA gene cloning library of dairy goat intestinal microorganism was constructed and data analysis was performed. The results of the dilution curves and the diversity index showed that single AFB₁ or mixed with other toxins had a tendency to reduce the dairy goat intestinal microorganism,but there was no significant difference (P>0.05).However,the total number of intestinal flora decreased significantly after AFB₁ mixed with other toxins treatement (P<0.05). The Firmicutes,Bacteroidetes and Tenericutes were found as the main phylum in the intestines of dairy goats using the species annotation of the obtained OTUs representative sequence. According to LEfSe analysis,the combination of AFB₁,OTA and ZEN could significantly increase the abundance of Succinivibrionaceae,Phascolarctobacterium and Aeromonadales (P<0.05).The abundance of Holdemania and BF311 bacillus in control group were significantly higher than those of other groups (P<0.05). These results indicated that AFB₁ mixed with ZEN and OTA could influence the gut microbial community structure of dairy goat,and the effects of different mycotoxin combinations complied with the following pattern:AFB₁+OTA+ZEN > AFB₁+OTA > AFB₁+ZEN > AFB₁.

Key words: dairy goat; mycotoxin; intestinal microflora; 16S rRNA

中图分类号:

S826

范彩云, 苏娣, 李晓娇, 黄帅, 张养东, 郑楠, 刘光磊, 刘祯, 程建波. 黄曲霉毒素B₁、赭曲霉毒素A和玉米赤霉烯酮联合对奶山羊肠道微生物的影响[J]. 《中国畜牧兽医》, 2017, 44(12): 3418-3425.

FAN Cai-yun, SU Di, LI Xiao-jiao, HUANG Shuai, ZHANG Yang-dong, ZHENG Nan, LIU Guang-lei, LIU Zhen, CHENG Jian-bo. Effects of Aflatoxin B₁ Individual,Binary or Tertiary Mixed with Ochratoxin A and Zearalenone on Intestinal Microorganism of Dairy Goat[J]. , 2017, 44(12): 3418-3425.

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黄曲霉毒素B₁、赭曲霉毒素A和玉米赤霉烯酮联合对奶山羊肠道微生物的影响

Effects of Aflatoxin B₁ Individual,Binary or Tertiary Mixed with Ochratoxin A and Zearalenone on Intestinal Microorganism of Dairy Goat

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