【作者】 张凯；

【导师】 董双林； 田相利；

【作者基本信息】 中国海洋大学 ， 水生生物学， 2014， 博士

【摘要】 本文根据生物学和生态学原理，对三疣梭子蟹、凡纳滨对虾与菲律宾蛤仔混养围隔的有机碳收支以及细菌生产力与代谢功能进行了比较研究，包括梭子蟹-对虾二元混养（CS）、梭子蟹-蛤仔二元混养（CB2）、对虾-蛤仔二元混养（SB2）、梭子蟹-对虾-蛤仔三元混养（菲律宾蛤仔以四个不同密度与虾、蟹混养，分别命名为CSB1、CSB2、CSB3和CSB4），以梭子蟹单养作为对照（C）。研究内容包括：三疣梭子蟹、凡纳滨对虾与菲律宾蛤仔混养系统有机碳收支、有机碳库储量、底泥有机碳垂直分布特征、细菌生产力以及微生物群落功能多样性研究。旨在阐明三疣梭子蟹不同养殖模式下有机碳收支以及细菌生产力与代谢功能，总结出该种养殖模式的特点及其带来的效益，为三疣梭子蟹养殖业的健康、可持续发展提供参考。主要结果如下：1三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔混养系统有机碳收支的研究利用海水池塘陆基实验围隔，研究了三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养系统有机碳的收支、利用率及养殖效果等。结果表明：（1）适宜的三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔三元混养，能有效提高三疣梭子蟹养殖的综合效益。（2）浮游植物初级生产是围隔中有机碳的主要来源，占总输入量的46.13%⁷9.14%；其次为投喂饲料带入的有机碳，占总输入量的18.07%⁵1.37%。有机碳的支出主要为生物和底泥呼吸作用，占总支出量的50.84%⁷7.30%；其次为底泥积累，占总支出量的15.84%²5.39%。（3）各处理比较，三元混养CSB3组有机碳的利用率显著提高，比其它处理提高了1.80%⁶.14%。（4）综合养殖效果和各系统有机碳利用率结果表明，在本研究条件下，虾、蟹、贝混养最优比例为三疣梭子蟹6ind/m2，凡纳滨对虾45ind/m2，菲律宾蛤仔30⁶0ind/m2。2三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养系统有机碳库储量研究利用海水池塘陆基实验围隔，研究了三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养系统有机碳库储量，结果表明：（1）各处理POC和DOC分别为（1.71±0.05）^4.68±0.46mg C/L和（5.60±0.58）^18.44±1.81mg C/L。其中，浮游植物有机碳波动范围为（0.08±0.02）^1.53±0.10mg C/L，平均为0.64mg C/L；浮游动物有机碳波动范围为（10.58±1.71）^{173.70±13.68}μgC/L，平均为46.59μgC/L；细菌有机碳波动范围为（0.10±0.04）^1.46±0.05mg C/L，平均为0.41mg C/L。（2）各处理DOC和POC分别占总有机碳（TOC）的77.95%⁸1.89%和18.11%²2.05%。 POC中，腐质颗粒有机碳占TOC的6.92%¹3.85%；其次为浮游植物碳，占TOC的3.67%⁷.33%；浮游细菌碳和浮游动物碳分别占TOC的2.53%⁴.59%和0.25%⁰.61%。3三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养系统底泥有机碳储量及垂直分布特征的研究利用海水池塘陆基实验围隔，研究了三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养系统底泥有机碳储量及垂直分布特征进行研究。结果表明：（1）不同养殖模式底泥有机碳、微生物碳及易氧化碳含量均随深度增加而逐渐降低。（2）实验期间各处理底泥有机碳、微生物碳、易氧化碳波动范围分别为0.85².91mg/g、2.75×10-4⁷.40×10-2mg/g和0.23¹.14mg/g，微生物碳、易氧化碳占底泥有机碳的比例波动范围为0.78%¹.33%和35.70%⁴0.39%。（3）蟹单养和虾贝混养处理底泥有机碳稳定性较差，适宜密度虾蟹贝三元混养能提高底泥微生物活性。4三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养模式的微生物群落功能多样性的研究应用BIOLOG生态微板研究了三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔不同混养模式的微生物群落功能多样性，结果表明，不同混养生态系统微生物群落代谢功能在整个养殖期间呈现动态变化趋势，对底物碳源的利用数量和利用强度也具有较大差异：（1）随着养殖时间的推移，三元混养处理CSB3水体和底泥微生物碳代谢活性显著高于其它养殖模式（P<0.05），而蟹单养处理C的水体和底泥微生物碳代谢活性均为最低（P<0.05）。（2）不同处理水体和底泥微生物群落对碳源种类利用差异随时间分化显著。实验后期，三元混养处理CSB1、CSB3、CSB4水体微生物群落和CSB3底泥微生物群落对6类碳源的利用强度总体上均显著高于梭子蟹单养模式（P<0.05）。（3）PCA分析表明，实验期间系统微生物群落一直处于不断演替和变化之中，最终，三元混养处理微生物群落组成和代谢特征已与其他处理显著不同。（4）各处理比较，实验后期，三元混养处理CSB3、CSB4水体微生物群落和CSB3底泥微生物群落功能多样性总体上均显著高于梭子蟹单养模式（P<0.05）。综上，三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔合理混养（CSB3）可以显著提高水体和底泥微生物群落的碳代谢活性和功能多样性，其次为处理CSB4。即在本研究条件中，蟹、虾、贝单元混养最优比例为三疣梭子蟹6ind/m2，凡纳滨对虾45ind/m2，菲律宾蛤仔30⁶0ind/m2。5三疣梭子蟹、凡纳滨对虾、菲律宾蛤仔混养系统浮游细菌生产力的研究利用陆基实验围隔，采用原位培养法，对比研究三疣梭子蟹、凡纳滨对虾和菲律宾蛤仔混养系统中细菌生产力的状况。结果表明，细菌生产力波动范围为（157.02±21.19）^{766.94±30.45}μgC/（L·d），平均值为377.57μgC/（L·d）。总体上，各处理组细菌生产力随时间呈现先升高后降低的趋势。实验前期，各处理细菌生产力相近；实验中后期，三元混养处理CSB3、CSB4的细菌生产力显著高于其他处理（P<0.05）。冗余分析发现，实验期间，细菌生产力、生物量与环境因子具有很好的相关性；细菌生产力的主要影响因子为DOC、POC、叶绿素a和浮游植物初级生产力；细菌数量的主要影响因子为NH4-N、NO2-N、NO3-N和PO4-P。更多还原

【Abstract】 In this thesis, total organic carbon budgets and bacterial productivity andmetabolism of polyculture system of Portunus trituberculatus, Litopenaeus vannameiand Ruditapes philippinarum were studied with land-based enclosures, including totalorganic carbon budgets, organic carbon storage, sediment organic carbon storage andvertical distribution, bacterial productivity and functional diversity of microbialcommunities of polyculture system of Portunus trituberculatus, Litopenaeusvannamei and Ruditapes philippinarum. There were8culture systems, monocultureof Portunus trituberculatus (C), polyculture of Portunus trituberculatus andLitopenaeus vannamei (CS), polyculture of Portunus trituberculatus and Ruditapesphilippinarum (CB2), polyculture of Portunus trituberculatus, Litopenaeus vannameiand Ruditapes philippinarum (CSB1, CSB2, CSB3and CSB4), polyculture ofLitopenaeus vannamei and Ruditapes philippinarum (SB2). The main results aredetailed as follows:1Total organic carbon budgets of polyculture system of Portunus trituberculatus,Litopenaeus vannamei and Ruditapes philippinarumThe budget of total organic carbon (TOC) was studied with land-basedenclosures with different polyculture systems of Portunus trituberculatus,Litopenaeus vannamei and Ruditapes philippinarum. The results showed that thebenefit of Portunus trituberculatus was significantly improved in the polyculturesystem of Portunus trituberculatus, Litopenaeus vannamei and Ruditapesphilippinarum. Primary production of phytoplankton was the main TOC (accountingfor46.13%~79.14%) inputs in the enclosures while the feed provided 18.07%~51.37%of TOC inputs. Among the output items, the major output of TOCwas the biological and sediment respiration, which accounted for50.84%~77.30%ofTOC outputs. The accumulation in sediment compromised15.84%~25.39%of TOCoutputs. The highest utilization efficiency of TOC of polyculture systems was CSB3system, which improved1.80%~6.14%than the other systems. The treatment givingthe best culturing benefit was crabs at6ind/m2, shrimps at45ind/m2and clams at30~60ind/m2.2Storage of the organic carbon of polyculture system of Portunus trituberculatus,Litopenaeus vannamei and Ruditapes philippinarumThe storage of organic carbon was studied with land-based enclosures withdifferent polyculture systems of Portunus trituberculatus, Litopenaeus vannamei andRuditapes philippinarum. The results showed that the storage of dissolved organiccarbon(DOC) and particulate organic carbon(POC) pools(mgC/L) ranged from(1.71±0.05) to (4.68±0.46) and (5.60±0.58) to (18.44±1.81), respectively. Organiccarbon storage of phytoplankton, zooplankton and bacteria ranged from (0.08±0.02)to (1.53±0.10)mg C/L,(10.58±1.71) to (173.70±13.68)μgC/L and (0.10±0.04) to(1.46±0.05)mg C/L, respectively. The ratios of DOC and POC to total organiccarbon(TOC) were77.95%~81.89%and18.11%~22.05%, respectively. The ratios ofdetritus organic carbon(DC), phytoplankton organic carbon(PBC), bacterial organiccarbon(BC) and zooplankton organic carbon(ZBC) were6.92%~13.85%,3.67%~7.33%,2.53%~4.59%and0.25%~0.61%, respectively.3Sediment organic carbon storage and vertical distribution of polyculturesystem of Portunus trituberculatus, Litopenaeus vannamei and RuditapesphilippinarumThe sediment organic carbon storage and vertical distribution was studied withland-based enclosures with different polyculture systems of Portunus trituberculatus,Litopenaeus vannamei and Ruditapes philippinarum. The results showed thatSediment Organic Carbon(SOC), Microbial Biomass Carbon(MBC) and ReadilyOxidizable Carbon (ROC) decreased with the depth of sediment layers. The contentsof SOC, MBC and ROC changed with the range of0.85~2.91mg/g, 2.75×10-4~7.40×10-2mg/g and0.23~1.14mg/g, respectively. The proportion of MBCand ROC changed with the range of0.78%~1.33%and35.70%~40.39%respectively.SOC of C and SB2treatments got lower stability while the polyculture of Portunustrituberculatus, Litopenaeus vannamei and Ruditapes philippinarum could improvemicrobial communities activity in sediment.4Variation of functional diversity of microbial communities in the polyculturesystem of Portunus trituberculatus, Litopenaeus vannamei and RuditapesphilippinarumFunctional diversity of water microbial communities were examined usingthe Biolog method in land-based enclosures with different polyculture systems ofPortunus trituberculatus, Litopenaeus vannamei and Ruditapes philippinarum.The results, including the carbon utilization, diversity indexes (Shannon, Simpson,McIntosh, S-E), and principal component analysis (PCA) with carbon sources,showed that microbial community functional diversity were significantlydifferent among8systems throughout the experiment, especially in Septemberwhen compared with in July.(1) In the later stage of the experiment, metabolicactivity of the microbial communities in CSB3were significantly higher than theother treatments while C was the lowest treatment (P<0.05).(2) The microbialcommunities in the water and sediment of different treatments of changedsignificantly on the use of carbon sources over time. In the later stage of theexperiment, the microbial communities in the water of CSB1, CSB3and CSB4treatments and sediment of CSB3treatment had significantly higher intensity thanC treatment on the using of carbon source (P<0.05).(3) PCA analysis showedthat microbial community functional diversity of CSB1, CSB2, CSB3and CSB4treatments were significantly different with the other treatments throughout theexperiment.(4) In the later stage of the experiment, the microbial communityfunctional diversity of microbial communities in the water of CSB3and CSB4treatments and sediment of CSB3treatment was significantly higher than Ctreatment. Our results indicated that the microbial communities in CSB3hadsignificantly higher functional diversity (P<0.05) than any other treatments. This study may provide a novel strategy for the culture of Portunus trituberculatus.5Bacterial Productivity of polyculture system of Portunus trituberculatus,Litopenaeus vannamei and Ruditapes philippinarumBacterial productivity (BP) was studied in situ with land-based enclosureswith different polyculture systems of Portunus trituberculatus, Litopenaeusvannamei and Ruditapes philippinarum. The results showed that BP ranged from(157.02±21.19) to (766.94±30.45)μgC/(L·d) with an average of377.57μgC/(L·d).The BP showed the same trend in general, increasing atbeginning and then decreasing. At the early stage of the study, BP of differenttreatments showed the similar trend. While at the middle and later stage, BP ofCSB3and CSB4were significantly higher than the other treatments(P<0.05).Redundancy analysis showed that was BP and bacterial number(BN) were closelyrelevant to environmental factors. The most significant environmental variableswere dissolved organic carbon(DOC), particulate organic carbon(POC),chlorophyll a and phytoplankton productivity(PP) with BP while NH4-N, NO2-N,NO3-N and PO4-P with BN, respectively.更多还原