醋酸棉酚对绵羊瘤胃液真菌多样性的影响

Effects of Gossypol Acetic Acid on Rumen Fungi Diversity of Sheep

本试验旨在运用高通量测序方法研究醋酸棉酚对绵羊瘤胃液真菌多样性的影响。选取8只健康、体重为(49.13±4.70)kg、安装有永久性瘤胃瘘管的哈萨克羊母羊,随机分为2组,分别为对照组和试验组,每组4只。试验期对照组绵羊饲喂基础饲粮;试验组绵羊第1~26天在基础饲粮中添加600 mg/(d只)醋酸棉酚,在第27~47天醋酸棉酚添加量增加至1200mg/(d只)。采集试验期正试期第1、20、37和47天瘤胃液样品,运用Illumina Hiseq平台测序分析绵羊瘤胃液真菌变化。结果显示:1)由样品物种稀释曲线和累积曲线可以看出,测序深度已经覆盖了瘤胃液样品中大部分的真菌。2)第1、20、47天,试验组与对照组相比瘤胃液真菌的Alpha多样性指数差异不显著(P>0.05);第37天时,试验组瘤胃液真菌的Shannon和Simpson指数显著高于对照组(P<0.05)。3)主坐标分析图显示,试验组与对照组瘤胃液真菌样品聚集在一起。4)在属水平上,第20和37天,试验组瘤胃液真菌区系中毛霉菌属(Mucor)相对丰度显著高于对照组(P<0.05)。第47天,试验组绵羊瘤胃液真菌区系中Metschnikowia相对丰度显著高于对照组(P<0.05)。综上所述,低浓度醋酸棉酚对绵羊瘤胃液真菌多样性无明显影响,醋酸棉酚浓度增加至1200mg/(d只)时,Simpson和Shannon指数显著升高;醋酸棉酚对绵羊瘤胃液真菌主要优势菌属组成无显著影响。

This study was aimed to evaluate the effects of gossypol acetic acid on rumen fungi diversity of sheep by high-throughput sequencing. Eight healthy Kazakh female sheep weighted( 49.13±4.70) kg with permanently rumen fistula were randomly divided into control group and trial group,each group contained 4 sheep. Sheep in the control group were fed a basal diet;and sheep in the trial group were fed the basal diet supplemented with 600 mg/d gossypol acetic acid per sheep during 1 to 26 days and 1 200 mg/d gossypol acetic acid per sheep during 27 to 47 days. Rumen fluid samples at days 1,20,37 and 47 were collected to determined fungi communities by Illumina Hiseq platform sequencing. The results showed as follows: 1) rarefaction curve and accumulation curve of sample species showed that all samples were reached the saturation phase,indicating depth of sequencing was reached. 2) On days 1,20 and 47,there was no significant effect on Alpha diversity of fungi in rumen fluid between control group and trial group( P > 0. 05);on day 37,the Shannon and Simpson indexes in rumen fluid of trial group were significantly higher than those of the control group( P<0.05). 3) The principal coordinate analysis diagram showed that rumen fluid fungi samples in control group and trial group clustered together. 4) At the genus levels,on days 20 and 37,the relative abundance of Mucor in rumen fluid fungi flora of trial group was significantly higher than that of the control group( P < 0.05);on day 47,the relative abundance of Metschnikowia in rumen fluid fungi flora of trial group was significantly higher than that of the control group( P<0.05). In summary,the lowconcentration of gossypol acetic acid has no significant effect on fungi diversity of sheep, when gossypol acetic acid concentration increases to1 200 mg/d per sheep,the Simpson and Shannon indexes significantly increase;gossypol acetic acid has no effect on predominant fungi composition in rumen fluid of sheep.