不同养殖模式下对虾早期死亡综合症致病菌的动态变化

[目的]为指导南美白对虾高位池的科学养殖提供理论依据。[方法]选取2种养殖模式的南美白对虾高位池塘为试验对象,通过周期性的水样采集研究了虾池水体细菌数量、弧菌数量以及弧菌中非副溶血弧菌和副溶血弧菌比例变化规律与早期死亡综合症发生的相关性。[结果]试验虾池在养殖初期弧菌数量较低,养殖过程中弧菌数量与细菌总数的消长趋势基本一致。当虾池中细菌总数高于2.5×104cfu/ml,弧菌数量高于1.5×103cfu/ml,非副溶血弧菌与副溶血弧菌的比例超过10∶1,虾池内极易发生早期死亡综合症。[结论]该研究结果可用于进行对虾健康养殖的风险评估。     

江苏沿海河蟹育苗水体中细菌种群数量变化

对江苏沿海河蟹育苗水体中异养菌和弧菌数变化进行了测定 ,研究了水体中细菌种群数量及河蟹幼体体内异养菌变化情况。结果表明 ,在河蟹育苗水体中 ,总异养菌数和弧菌数都随着幼体发育阶段的发展呈现出稳定增高趋势 ,总异养菌数从Z1期的 3.3× 10 3 cfu/ml上升至M期的 5 .2× 10 8cfu/ml;弧菌数从 7.8× 10 2 cfu/ml上升至 5 .2× 10 6cfu/ml。幼体体内的异养菌数量表现为由高而低的趋势 ,总体水平在 10 3 ～ 10 5cfu/g之间。育苗水体中异养菌组成主要包括弧菌属、假单胞菌属、发光杆菌属、芽孢杆菌属、黄杆菌属、产碱杆菌属等     

工厂化育苗养殖体系内细菌数量动态变化研究

以异养菌、弧菌、副溶血弧菌为监测对象,对中国对虾的工厂化育苗进行实时监测,探寻育苗过程中细菌的动态变化规律,为对虾健康养殖提供科学依据。     

不同密度青蛤养殖塘异养细菌和弧菌的数量变化

研究了青蛤(Cyclina sinensis)养殖环境中异养细菌数量和弧菌数量的动态变化,以探讨其与养殖密度的相关性和样本间相关性。结果表明:实验周期内各养殖塘水体中异养细菌总数呈现不同的变化趋势且差异不显著(P>0.05),而弧菌数量变化趋势较为一致,9月份与其他月份水样中弧菌数量差异较为显著(P<0.05)。各塘底泥中异养细菌总数变化和弧菌数量变化差异均不显著,而不同密度养殖塘底泥中弧菌数量变化差异显著(P<0.05)。不同采样时间同一塘的青蛤腔液异养细菌总数差异极显著(P<0.01),而弧菌数量差异较为显著(P<0.05)。样本间、样本与养殖密度间相关性分析发现,水体中异养细菌总数变化与弧菌数量变化呈显著正相关(r=0.335,P<0.05);底泥中异养细菌总数变化与弧菌数量变化呈正相关且相关性极显著(r=0.587,P<0.01);底泥和水体中异养细菌总数和弧菌数量变化与养殖密度相关性均不显著(P>0.05);青蛤腔液异养细菌总数变化与养殖密度相关性极显著(r=-0.189,P<0.01),弧菌数量变化与养殖密度相关性极显著(r=-0.123,P<0.01)。     

海参养殖池塘异养菌与弧菌数量变化特征分析

本研究对海参养殖池塘水体中的异养菌、弧菌数量变化进行监测,结果表明,表层水体异养菌数量为4.4×103~2.3×105 CFU/mL,中层水体异养菌数量为7.95×103~1.60×105CFU/mL,底层水体异养菌数量为5.0×103~1.6×105CFU/mL;表层水体弧菌数量为0~7.85×103 CFU/mL,中层水体弧菌数量为0~1.8×103 CFU/mL,底层水体弧菌数量为0~1.32×104 CFU/mL。池塘水环境中的异养菌总数和弧菌数均呈现出较为明显的季节变化,春季、夏季处于较高水平,秋季、冬季处于较低水平。其中,弧菌数量受温度影响较大,冬季达到最低值,随着养殖池塘水温的回升而逐渐升高。     

凡纳滨对虾循环水养殖系统弧菌消长研究

以鉴别性培养基TCBS琼脂采用平板菌落计数法对凡纳滨对虾循环水养殖系统水体中弧菌的消长进行了监测.结果表明,循环水养殖系统中主要存在非O1群霍乱弧菌、副溶血性弧菌、梅氏弧菌、溶藻弧菌和普通变形杆菌等病原细菌;与传统换水养殖模式相比,循环水养殖系统对非O1群霍乱弧菌、溶藻弧菌和普通变形杆菌控制效果更好,对副溶血性弧菌和梅氏弧菌控制效果相当;在整个养殖过程中,循环水养殖系统弧菌和普通变形杆菌的数量均低于对虾发病的阈值(104 CFU/mL);循环水养殖模式和传统换水养殖模式的水体pH值均在7～10范围内波动,但仅发现非O1群霍乱弧菌和副溶血性弧菌的消长与pH值波动有一定相关性.     

高位虾池水体细菌和弧菌的数量变化及影响因素

研究了位于广东湛江东海岛北寮村和庵里村的4个凡纳滨对虾（Penaeus vannamei）高位池养殖过程水体中的可培养异养细菌和弧菌的数量变化及其影响因素.结果表明：1）北寮村虾池BL1和BL2的可培养异养细菌数量变化均呈现中间高两头低、后期多于初期的单峰变化趋势;BL1的弧菌数量变化趋势和可培养异养细菌的变化趋势相似,而BL2则是前期稍微升高而后下降,后期又呈升高态势,峰值出现在养殖末期;水源地BL3的可培养异养细菌数量变化为初期平缓,后期明显多于初期,大体呈上升态势;而BL3弧菌数量变化趋势则比较平缓.2）庵里村发病的虾池AL1的可培养异养细菌和弧菌数量呈现出单峰变化趋势,在养殖中途爆发病害时,可培养异养细菌数量高出未发病的正常池2个数量级;庵里村虾池AL2的可培养异养细菌数量变化趋势也是中间高两头低,而弧菌的数量变化则是无规律的波动;水源地AL3的可培养异养细菌数量变化大体呈下降趋势,弧菌数量放养初期最高.3）高位虾池的可培养异养细菌和弧菌数量虽多于水源地,但差异不显著,同一个位点,细菌和弧菌数量呈现同步增多或减少.虾场水体的可培养异养细菌和弧菌数量与多数环境因子之间相关性不显著,这与虾池养殖生态系统受人为因素影响较大有关,同时反映了高位虾池生态系统受多种因子作用的复杂性.     

1株乳酸菌在凡纳滨对虾养殖中的应用效果

利用从对虾消化道分离出的1株乳酸菌优势菌株HN3,分别对哈维氏弧菌和副溶血弧菌进行体外拮抗实验,同时通过乳酸菌拌料投喂的方式开展凡纳滨对虾的养殖试验。结果表明:乳酸菌液及中和酸性后的胞外液均对2种弧菌有显著的抑制效果(P0.05);乳酸菌拌料投喂能显著提高对虾的养殖存活率、日增体质量和特定增长率(P0.05);对虾消化道和养殖水体中的乳酸菌和总细菌数量大幅增加,弧菌数量显著降低(P0.05),其中,以乳酸菌添加剂量4×106~5×106cfu·g-1的综合养殖效果最佳。此株乳酸菌适应对虾消化道的环境并能定植,对抑制对虾消化道弧菌数量、优化体内菌群组成和改善养殖环境有重要的作用。     

海湾扇贝幼体期流行性弧菌病的研究

从辽宁兴城育苗场和辽宁省海洋水产研究所育苗室患面盘解体的海湾扇贝Argopecten irradians Lamarck幼体中分离出6株菌,经感染试验表明,其中3株菌B01、Y01和Y02为病原菌。根据菌体形态及生理、生化测试,B01鉴定为需钠弧菌Vibrio natriegen,Y01和Y02具有相同的形态、生理、生化特征,鉴定为弧菌属Vibrio。需钠弧菌对庆大霉素、痢特灵、菌必治、诺氟沙星、氯霉素、环丙沙星、多粘菌素B敏感;Y01和Y02菌对诺氟沙星、菌必治、氯霉素、复方新诺明、环丙沙星、磺胺异嗯唑敏感。对海湾扇贝育苗水体中细菌总数和弧菌数量的监测结果表明,幼体发生病变之前的水体中,细菌和弧菌的密度高于正常育苗水体的。文中还对病原菌的传播途径等问题进行了讨论。     

副溶血弧菌噬菌体的吸附材料选择及其吸附和洗脱作用

[目的]选择副溶血弧菌噬菌体的适宜吸附剂,并研究其吸附和洗脱作用。[方法]比较了滑石粉+硅藻土(3∶1)、活性炭、蒙脱石、玻璃纤维4种吸附剂对副溶血弧菌噬菌体的吸附固定化效果以及洗脱效果。[结果]蒙脱石和玻璃纤维对噬菌体的吸附效果好。通过单因素试验和正交试验研究蒙脱石用量、吸附时间、吸附液p H对噬菌体固定化的影响,结果发现当蒙脱石用量为0.5 g,吸附时间为1 h,吸附液p H为6.0时,吸附效果最佳。[结论]将副溶血弧菌噬菌体制成固定化制剂后分别作用于养殖动物和养殖水体,可以达到对养殖动物进行生物防治和改善水体环境的目的。     

Study on the Quantity Dynamic Changes of Heterobacteria and Vibrios in Larvae Industrialized Culture System

[Objective] The dynamic change of heterobacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp.[Method] The heterobacteria,vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized culture system.[Result] The heterobacteria,vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in nauplius,then their number would increase with growth. During whole rearing period,both heterobacteria in larvae,vibrios in water would increase by one order of magnitude,while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude.There were many heterobacteria and vibrios but few vibrio parahaemolyticus in living bait.The correlation coefficients between larvae and heterobacteria and vibrios in water were 0.704 and 0.840 in culture system respectively,while the correlation among heterobacteria,vibrios in living bait and larvae,water were weak or negative.[Conclusion] There was a dynamic relation between water and larvae in rearing period,and restrictly control of culture condition would restrain the occurrence of disease caused by vibrio parahaemolyticus,besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.     

工厂化育苗养殖体系内细菌数量动态变化研究(英文)

[Objective] The dynamic change of heterobacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp.[Method] The heterobacteria,vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized culture system.[Result] The heterobacteria,vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in nauplius,then their number would increase with growth. During whole rearing period,both heterobacteria in larvae,vibrios in water would increase by one order of magnitude,while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude.There were many heterobacteria and vibrios but few vibrio parahaemolyticus in living bait.The correlation coefficients between larvae and heterobacteria and vibrios in water were 0.704 and 0.840 in culture system respectively,while the correlation among heterobacteria,vibrios in living bait and larvae,water were weak or negative.[Conclusion] There was a dynamic relation between water and larvae in rearing period,and restrictly control of culture condition would restrain the occurrence of disease caused by vibrio parahaemolyticus,besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.     

Study on the Quantity Dynamic Changes of Heterobacteria and Vibrios in Larvae Industrialized Culture System

[Objective] The dynamic change of heterebacteria and vibrios in larvae industrialized culture system was studied to provide scientific reference for healthy cultivation of shrimp. [Method] The hcterobacteria, vibrios and pathogenic vibrio parahaemolyticus were monitored in larvae industrialized cul-ture system. [Result] The heterobacteria, vibrios and pathogenic vibrio parahaemolyticus were the most in fertilized eggs of shrimp but the least in naupli-us, then their number would increase with growth. During whole rearing period, both heterobscteria in larvae, vibrios in water would increase by one order of magnitude, while both vibrios in larvae and heterobacteria in water would increase by two orders of magnitude. There were many heterohacteria and vibri-os but few vibrio parahaemolyticus in living bait. The correlation coeffcients between larvae and heterobacteria and vibrios in water were 0.704 and 0.840 in culture system respectively, while the correlation among heterobacteria, vibrios in living bait and larvae, water were weak or negative. [Conclusion] There was a dynamic relation between water and larvae in rearing period, and restrictly control of culture condition would restrain the occurrence of disceae caused by vibrio parahaemolyticus, besides that bacteria number in bait was not obviously correlated with bacteria nubmer in culture system.     

副溶血性弧菌的检测研究

[目的]对toxR进行副溶血性弧菌特异性检测,探讨toxR靶基因能否准确检测副溶血性弧菌,从而消除其他研究者对该基因特异性的疑虑。[方法]采用SN/T1870—2016中副溶血性弧菌toxR的引物探针对副溶血性弧菌和其亲缘关系接近的弧菌标准菌株进行检测。[结果]采用实时荧光PCR方法证实该引物探针只能扩增出副溶血性弧菌,其他弧菌诸如溶藻弧菌、创伤弧菌、霍乱弧菌等未得到扩增。[结论]证实SN/T1870—2016中toxR的引物探针特异性高。该研究可为各检测机构提供数据支持,为副溶血性弧菌的检测与研究奠定更为坚实的基础。     

致病性副溶血性弧菌特异性三重PCR检测方法研究

[目的]建立同时检测致病性副溶血性弧菌gyrase、tdh、toxR基因的三重PCR快速检测方法。[方法]用3种基因的特异性引物分别对副溶血性弧菌ATCC33847的模板DNA进行单一扩增,找到各自引物最佳扩增条件;再用3种引物同步对模板DNA进行扩增,通过优化引物浓度、引物间比例以及退火温度,建立最佳扩增体系。[结果]在最佳三重PCR反应条件下,gyrase、tdh和toxR能同时扩增出清晰条带,大小分别为91、269和368 bp。[结论]该研究为致病性副溶血性弧菌的快速检测提供了一种新的技术方法。     

利用PCR技术快速检测水产品中副溶血性弧菌

[目的]对副溶血性弧菌的检测进行研究。[方法]采用聚合酶链式反应（PCR）,以标准菌株（单增李斯特菌、哈氏弧菌）作为阴性对照对3株副溶血性弧菌进行特异性扩增,扩增引物采用副溶血性弧菌如基因中的tlh-3和tlh-4,同时利用PCR对人工污染的白对虾中的副溶血性弧菌进行检出限测定。[结果]结果表明,通过PER扩增,副溶血性弧菌在约449bp处扩增出特异性条带,单增李斯特菌、哈氏弧菌均未出现任何条带,扩增片段表现出极好的特异性。菌液稀释到3．3×10^2cfu／ml时,将其污染到白对虾中作PCR仍可扩增出目的片断。[结论]该研究表明PER检测方法快捷、特异性好、敏感性高,可以作为副溶血性弧菌辅助检测方法。     

文蛤生物体及内脏中弧菌的研究

[目的]鉴定文蛤体内弧菌的类型及致病性强度,并筛选治疗这些病原菌的最佳药物。[方法]用TCBS培养基从文蛤生物体内分离出12株弧菌,对其进行致病性试验,对致病性较强的3株弧菌进行鉴定及药物敏感试验。[结果]该3株菌分别为河流弧菌H04（Vibrio fluvialis）,H06副溶血弧菌（Vibrio parahaemolyticus）,H11创伤弧菌（Vibrio vulnificus）,其对环丙沙星、先锋赛高度敏感,对青霉素G、先锋霉素V有耐药性,临床上可以首选环丙沙星、先锋赛作为治疗这些病原菌的药物,其次是头孢曲松、氟哌酸、丁胺卡那、四环素、庆大霉素等。[结论]结果为文蛤疾病的防治提供了理论依据。     

副溶血弧菌对南美白对虾生理生化指标的影响

在自然条件下从患红体病的养殖南美白对虾肝脏内分离出病原菌——副溶血弧菌，将该菌通过肌肉注射的方式，对南美白对虾做感染实验，以探讨副溶血弧菌对南美白对虾生理生化指标的影响。注射弧菌后南美白对虾死亡率与时间、浓度呈正相关，与半致死剂量呈负相关；虾体肌肉、肝脏组织的超氧化物歧化酶（SOD）、溶菌酶（LSZ）酶活性随弧菌浓度的增加呈下降的趋势，而过氧化物酶（POD）酶活性呈上升趋势，这可能是应激反应；酯酶同工酶（EST）酶带表现出缺失，苹果酸脱氢酶同工酶（MDH）具有明显的组织特异性，并且两种酶在肌肉组织中表达最强。副溶菌弧菌感染后破坏了虾体的免疫系统，导致免疫机能的损坏，防病、抗病能力下降，进一步导致虾体患病，甚至死亡。     

副溶血弧菌对南美白对虾生理生化指标的影响

在自然条件下从患红体病的养殖南美白对虾肝脏内分离出病原菌——副溶血弧菌，将该菌通过肌肉注射的方式，对南美白对虾做感染实验，以探讨副溶血弧菌对南美白对虾生理生化指标的影响。注射弧菌后南美白对虾死亡率与时间、浓度呈正相关，与半致死剂量呈负相关；虾体肌肉、肝脏组织的超氧化物歧化酶（SOD）、溶菌酶（LSZ）酶活性随弧菌浓度的增加呈下降的趋势，而过氧化物酶（POD）酶活性呈上升趋势，这可能是应激反应；酯酶同工酶（EST）酶带表现出缺失，苹果酸脱氢酶同工酶（MDH）具有明显的组织特异性，并且两种酶在肌肉组织中表达最强。副溶菌弧菌感染后破坏了虾体的免疫系统，导致免疫机能的损坏，防病、抗病能力下降，进一步导致虾体患病，甚至死亡。