幼龄朱鹮肠道微生物多样性与产胞外酶活力分析  

Intestinal Microbial Diversity and Enzyme Activities of Crested Ibis Nestlings

作　　者：王琦[1,2] 兰阿峰[1,2] 王华健 郭素芬 段文斌[3] 牛克胜 WANG Qi;LAN A-Feng;WANG Hua-Jian;GUO Su-Fen;DUAN Wen-Bin;NIU Ke-Sheng(College of Biological Science and Engineering,Shaanxi University of Technology,Hanzhong 723001;Qinba State Key Laboratory of Biological Resources and Ecological Environment,Shaanxi University of Technology,Hanzhong 723001;Shaanxi Hanzhong Crested Ibis National Nature Reserve,Yangxian 723300,China)

机构地区：[1]陕西理工大学生物科学与工程学院,汉中723001 [2]陕西理工大学秦巴生物资源与生态环境省部共建国家重点实验室(培育),汉中723001 [3]陕西汉中朱鹮国家级自然保护区管理局,洋县723300

出　　处：《动物学杂志》2024年第5期714-728,共15页Chinese Journal of Zoology

基　　金：陕西省科技厅项目(No.2015SZS-15-09)。

摘　　要：朱鹮(Nipponia nippon)是国家一级重点保护野生动物,属于世界上最濒危的鸟类之一,本研究通过对幼龄朱鹮肠道微生物的多样性及产酶活力进行研究,以期分离得到对朱鹮有益的微生物。本研究采用纯培养的方法获得幼龄朱鹮肠道微生物,通过革兰氏染色以及生理生化鉴定,结合16S rRNA基因扩增和序列分析进行鉴定。使用水解圈降解法筛选产淀粉酶、蛋白酶、纤维素酶和脂肪酶的菌株,最终获得254株幼龄朱鹮肠道细菌,隶属于2门9属。其中变形菌门(Proteobacteria)211株,占分离菌株总数的83.07%,优势菌属为埃希氏菌属(Escherichia)179株(70.47%)、柠檬酸杆菌属(Citrobacte)18株(7.09%)以及肠杆菌属(Enterobacter)14株(5.51%);厚壁菌门(Firmicutes)43株,占分离总数的16.93%,优势菌属为肠球菌属(Enterococcus)27株(10.63%)。菌株生理生化鉴定出的种属与各自的16SrRNA基因鉴定出的种属相一致。产酶活力分析结果显示,有244株产蛋白酶,19株产淀粉酶,10株产脂肪酶,4株产纤维素酶,分别占分离菌株总数的83.07%、7.48%、3.94%和1.57%。开展肠道微生物研究可为朱鹮饲养提供理论指导,挽救每年夏秋季节因肠道疾病死亡的朱鹮。因此,本研究期望能分离筛选出对朱鹮肠道有益的微生物菌株,对朱鹮肠道疾病、生长发育及种群恢复提供帮助。[Objectives]Crested Ibis Nipponia nippon is protected at the national level in China and is also one of the most critically endangered bird species globally.Unfortunately,the population of wild Crested Ibises has experienced a drastic decline due to environmental degradation.Research indicates that the gut microorganisms of Crested Ibises play vital roles in nutrient absorption,immune regulation and healthy growth.The objective of this study was to culturally analyze the intestinal microbiota of Crested Ibis nestlings for beneficial microorganisms.In this study,the gut microbial diversity and extracellular enzyme activity of Crested Ibises were examined to further understand their healthy development.[Methods]The intestinal microorganisms in Crested Ibis nestlings were obtained through pure culture methodology.The identification of the bacteria involved a combination of Gram staining,physiological and biochemical analysis,and 16S rRNA gene amplification and sequence analysis.A phylogenetic tree was subsequently constructed using 16S rRNA gene sequences of intestinal bacteria derived from Crested Ibis nestlings,including representative isolates and related species.Strains that produced amylase,protease,cellulase,or lipase were screened through hydrolytic cycle degradation.[Results]A total of 254 strains of bacteria were isolated from the fresh excreta of artificially-reared Crested Ibis nestlings,comprising 2 phyla and 9 genera(Table 2).Thirty-three gram-positive strains and 221 strains of gram-negative bacteria were identified(Fig.1).Among these strains,211 were identified as Proteobacteria,accounting for 83.07%of the total isolates.Among them,179 were Escherichia sp.,representing 70.47%of the total number of isolates,additionally,18 strains of Citrobacter sp.(7.09%),14 strains of Enterobacter sp.(5.51%),and 14 strains of Klebsiella sp.(5.51%)were also isolated.Of the total number of isolates,7.09%were identified as follows:fourteen strains of Enterobacter sp.(5.51%),4 strains of Klebsiella sp.(1.57%),4 strains o

关 键 词：幼龄朱鹮 肠道微生物 纯培养 16S rRNA基因 产胞外酶 

分 类 号：Q958[生物学—动物学]