黄雪敏,温崇庆,梁华芳,洪居恳,薛明

• 1:广东海洋大学水产学院

摘要(Abstract)：

【目的】揭示凡纳滨对虾(Litopenaeus vannamei)糠虾期健康和发病育苗池水体菌群结构差异。【方法】从健康和发病育苗池各采集3份水样,提取水样基因组DNA,利用Illumina Mi Seq测序技术对其进行16S高通量测序,分析菌群结构组成与多样性,并筛选指示健康状态的水体菌群。【结果】健康苗池水体菌群多样性和均匀度均显著高于发病苗池(P<0.05)。健康苗池中放线菌门(Actinobacteria)及其微杆菌科(Microbacteriaceae)、拟杆菌门(Bacteroidetes)及其圆杆菌科(Cyclobacteriaceae)和黄杆菌科(Flavobacteriacea)、β-变形菌纲(β-Proteobacteria)及其丛毛单胞菌科(Comamonadaceae)、庞微菌门(Verrucomicrobia)及其分类未定科(OTU9),均优势存在,且序列丰度显著高于发病池(P<0.01);相反,发病池中α-变形菌纲(α-Proteobacteria)及其红杆菌科(Rhodobacteraceae)和Stappia_f科、γ-变形菌纲(γ-Proteobacteria)及其交替单胞菌科(Alteromonadaceae)丰度均显著高于健康池(P<0.01)。属水平上,发病池红杆菌科4个分类未定属(OTU1、OTU6、OTU64、OTU123)、亚硫酸杆菌属(Sulfitobacter),以及交替单胞菌科交替单胞菌属(Alteromonas)丰度均显著高于健康池(P<0.01),而微杆菌科Limnoluna与Rhodoluna属和红杆菌科一个分类未定属(OTU4)丰度均显著低于健康池(P<0.01)。【结论】健康和发病糠虾期育苗池水体菌群结构和多样性存在显著差异,属水平上丰度差异显著的优势菌群可作为苗池健康状态指示菌群。

关键词(KeyWords)： 凡纳滨对虾;育苗池;健康状态;水体菌群;多样性;高通量测序;指示菌群

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(31502189,31372536)

作者(Author): 黄雪敏,温崇庆,梁华芳,洪居恳,薛明

参考文献(References)：

• [1]徐怀恕,杨学宋,李筠,等.对虾育苗期细菌病害的诊断与控制[M].北京:海洋出版社,1999:1-51;136-152.
• [2]VANDENBERGHE J,VERDONCK L,ROBLES-AROZARENA R,et al.Vibrios associated with Litopenaeus vannamei larvae,postlarvae,broodstock,and hatchery probionts[J].Applied and Environmental Microbiology,1999,65(6):2592-2597.
• [3]FAO.Health management and biosecurity maintenance in white shrimp(Penaeus vannamei)hatcheries in Latin America[R].FAO Fisheries Technical Paper No.450.Rome,2003.
• [4]ZHENG Y F,YU M,LIU Y,et al.Comparison of cultivable bacterial communities associated with Pacific white shrimp(Litopenaeus vannamei)larvae at different health statuses and growth stages[J].Aquaculture,2016,451:163-169.
• [5]DE SCHRYVER P,VADSTEIN O.Ecological theory as a foundation to control pathogenic invasion in aquaculture[J].ISME Journal,2014,8(12):2360-2368.
• [6]ZHANG D M,WANG X,XIONG J B,et al.Bacterioplankton assemblages as biological indicators of shrimp health status[J].Ecological Indicators,2014,38(38):218-224.
• [7]温崇庆,薛明,何红,等.两株对虾幼体弧菌病病原的分离和鉴定[J].微生物学通报,2008,35(3):346-352.
• [8]ZHENG Y F,YU M,LIU J W,et al.Bacterial community associated with healthy and diseased pacific white shrimp(Litopenaeus vannamei)larvae and rearing water across different growth stages[J].Frontiers in Microbiology,2017,8:1362.
• [9]黄永春.有效微生物菌群对养虾水体细菌生态和水质的影响[J].广东海洋大学学报,2009,29(1):44-48.
• [10]BLANCHETON J P,ATTRAMADAL K J K,MICHAUD L,et al.Insight into bacterial population in aquaculture systems and its implication[J].Aquacultural Engineering,2013,53(3):30-39.
• [11]XUE M,LIANG H F,HE Y Y,et al.Characterization and in-vivo evaluation of potential probiotics of the bacterial flora within the water column of a healthy shrimp larviculture system[J].Chinese Journal of Oceanology&Limnology,2016,34(3):484-491.
• [12]薛明,何瑶瑶,邱孟德,等.高通量测序分析凡纳滨对虾育苗期水体菌群结构特征[J].水产学报,2017,41(5):785-794.
• [13]张吕平,胡超群,罗鹏,等.抗生素对于对虾苗池水体细菌区系的影响[J].应用生态学报,2009,20(10):2551-2557.
• [14]WEN C Q,XUE M,LIANG H F,et al.Evaluating the potential of marine Bacteriovorax,sp.DA5 as a biocontrol agent against vibriosis in Litopenaeus vannamei,larvae[J].Veterinary Microbiology,2014,173(1/2):84-91.
• [15]ZHOU J Z,WU L Y,DENG Y,et al.Reproducibility and quantitation of amplicon sequencing-based detection[J].ISME Journal,2011,5(8):1303-1313.
• [16]XIONG J B,ZHU J L,ZHANG D M.The application of bacterial indicator phylotypes to predict shrimp health status[J].Applied Microbiology&Biotechnology,2014,98(19):8291-8299.
• [17]中华人民共和国国家标准.海洋监测规范第4部分:海水分析(GB17378.4-2007)[S].北京:中国标准出版社,2007.
• [18]温崇庆,何瑶瑶,薛明,等.高通量测序分析DNA提取引起的对虾肠道菌群结构偏差[J].微生物学报,2016,56(1):130-142.
• [19]KESSNER L,SPINARD E,GOMEZ-CHIARRI M,et al.Draft genome sequence of Aliiroseovarius crassostreae CV919-312,the causative agent of Roseovarius Oyster disease(formerly juvenile oyster disease)[J].Genome Announcements,2016,4(2):e00148-16.