施用微生物制剂对育蚌水体微生物区系的影响

研究了育蚌水体分别施用微生物制剂利水素与鱼虾菌乐后水样中微生物区系的变化。试验结果表明:施用利水素的水样革兰氏阴性杆菌和嗜水气单胞菌含量分别下降26.67%、20%,芽胞杆菌和表皮葡萄球菌含量分别上升13.33%、26.67%;施用鱼虾菌乐的水样革兰阴性杆菌和嗜水气单胞菌含量分别下降20%、13.33%,芽胞杆菌和表皮葡萄球菌含量分别上升6.67%、13.33%;未施用微生物制剂的水样革兰阴性杆菌和嗜水气单胞菌含量均上升13.33%,芽胞杆菌消失,新增肠杆菌和链球菌。施用微生物制剂利水素和鱼虾菌乐,可有效优化养殖水体微生物种群结构,改良养殖水质环境。     

养殖中后期高位池对虾水体微生物群落结构及水体理化因子

针对凡纳滨对虾(Litopenaeus vannamei)高位池越冬棚养殖模式,采集养殖中后期水样,分析水体微生物群落结构、优势菌种类和水体理化因子的动态变化。结果表明:越冬棚保温效果良好,池塘水温由搭棚前最低的20.6℃上升并维持稳定于26.5℃左右,可满足对虾正常生长需求;养殖中后期水体异养细菌数量在5.10×104~2.95×105cfu·m L~(-1)波动,菌群结构相对稳定,优势菌数量在养殖后期略有增加;嗜冷杆菌(Psychrobacter)、亚硫酸杆菌(Sulfitobacter)等在养殖中后期均为优势种,搭棚后黄杆菌(Flavobacteria)、生丝微菌(Hyphomicrobium)等形成优势;亚硝氮、硝氮和水温是影响养殖中后期水体微生物群落分布的主要理化因子。研究表明,搭建越冬棚有利于保持养殖池塘水环境的相对稳定,养殖中后期水体氮素水平的控制尤为重要。因此,为促进水中有益菌的生态优势,调控水体菌群结构和生态功能,建议在养殖中后期,尤其是搭建越冬棚前后合理使用益生菌制剂,净化水质,稳定生态环境。     

鲤肠道正常菌群的研究

本实验用稀释滴种的定量方法,对淡水养殖地中健康鲤肠道内的１０种菌群进行了定性、定量分析,并对结果进行统计学处理,得到鲤肠菌群的一些生理值。结果表明鲤肠道中需氧、兼性厌氧优势菌是气单胞菌和酵母菌,厌氧优势菌是拟杆菌。对不同时间、不同温度条件下同一养殖池中鲤肠道菌群的测定结果比较表明：葡萄球菌、假单胞菌差异显著（ｐ＜０．０５）,气单胞菌、大肠杆菌、需氧芽胞杆菌、酵母菌、乳酸杆菌、双歧杆菌、拟杆菌、梭状芽胞杆菌差异不显著（ｐ＞０．０５）,表明该８种菌群在鲤肠道中的菌数处于相对稳定状态,属于肠道正常菌群。     

芽胞杆菌对鱼池微生物群落代谢功能的影响

利用Biotog平板,以施用与不施用芽胞杆菌的鱼塘为研究对象,研究鱼塘水体与表泥不同空间的微生物群落代谢功能的差异。结果表明：芽胞杆菌提高了水体微生物群落代谢的平均活性,及对氨基酸类、胺类、羧酸类、聚合物及其它类碳源的利用能力,提高了水体微生物群落代谢的多样性（Shannon均度、Simpson指数、McIntosh均度差异显著）;两池表泥微生物群落代谢平均活性差异较小,对同一碳源的利用差异不大,芽胞杆菌显著提高了表泥的Simpson指数,但表泥的McIntosh指数显著低于对照池。总体而言,鱼塘施用芽胞杆菌,水体微生物群落代谢功能所受的影响大过表泥,芽胞杆菌通过提高水体微生物群落代谢活性,提高了水体微生物对有机污染物的降解能力,以此改善池塘的生态环境。     

OLAND生物脱氮系统中硝化菌群16S rDNA的DGGE分析

为了考察生物脱氮系统中硝化菌群（氨氧化菌和亚硝酸氧化菌）的种群多样性及硝化菌群随溶解氧降低的种群变化规律,并建立一套行之有效的用于自养生物脱氮系统中功能微生物菌群的快速分子检测技术,采用DGGE（变性梯度凝胶电泳）分子检测技术对硝化菌群的16SrDNA的特异性PCR扩增产物进行了分析,结果表明：OLAND生物脱氮系统中氨氧化菌和亚硝酸氧化菌随溶解氧的降低表现出了不同的种群变化规律,氨氧化菌种群多样性受溶解氧的影响非常大,而非亚硝酸氧化菌的种群多样性比较单一,且不受溶解氧的影响。结合FISH（全细胞荧光原位杂交）分析结果表明,在OLAND限氧稳定运行后期,亚硝化单胞菌属（Nitrosomonas）是主要的氨氧化菌,占OLAND限氧亚硝化阶段反应器中总细菌数的72.5%左右。     

一株高效去除亚硝酸氮细菌的分离鉴定及其脱氮特性研究

【目的】从南美白对虾养殖塘中分离到高效去除亚硝酸氮的细菌, 对其分类和脱氮特性进行了研究。【方法】 以除亚硝酸氮为主要指标, 取养殖塘底层水样分离筛选菌株; 依据16S rRNA基因序列和生理生化特征初步鉴定菌株; 研究不同碳源、碳氮比、起始pH、温度、摇床转速和氯化钠浓度对反硝化除亚硝酸氮的影响, 并考查了菌株对硝酸氮和氨氮的利用情况。【结果】得到的菌株中菌株FP6活性最高, 初步鉴定菌株FP6属于地衣芽孢杆菌。菌株FP6的生长最适脱氮碳源为蔗糖, 菌株FP6去除亚硝酸氮有高活性的条件范围为: C/N值15?25、起始pH 7.0?10.0、温度20 °C?37 °C、摇床转速0?200 r/min和氯化钠浓度0?40 g/L。菌株FP6对硝酸氮和氨氮都有一定的去除能力, 利用硝酸氮时不积累亚硝酸氮。【结论】地衣芽孢杆菌FP6具有优良的除亚硝氮特性, 适宜的温度、pH和盐度范围较宽。     

水质净化酵母菌的分离筛选及鉴定

摘要:【目的】针对目前水产养殖专用优良菌种资源缺乏的现状,从养殖环境和养殖生物体中分离筛选具有水质净化功能的酵母菌,并对优良菌株进行鉴定。【方法】在低温和常温条件下从皮皮虾、南美白对虾肠道及养殖池底质活性污泥中分离具有水质净化功能的酵母菌,在模拟水体中对分离菌株的水质净化能力进行筛选,并对优良菌株采用形态、生理生化实验及5.8S rDNA ITS序列分析进行鉴定。【结果】从3种介质中共分离到酵母菌37株,其中常温分离16株,低温分离21株。水质净化实验结果表明,常温分离的16株酵母菌中有5株,低温分离的21株酵母菌中有6株对模拟水体中亚硝态氮和氨氮有显著的去除效果;其中低温分离的DN9和常温分离的CN6 48h能将10.64mg/L的亚硝态氮彻底转化,96h对630mg/L CODcr的去除率分别达52%和67%。DN9和CN6均产红色色素,经形态特征、生理生化特性及5.8S rDNA基因序列分析,鉴定菌株CN6为沼泽生红冬胞酵母(Rhodosporidium paludigenum),DN9为胶红酵母(Rhodotorula mucilaginosa)。【结论】红酵母DN9和CN6能有效去除养殖水体中的有机污染物和亚硝态氮,有望开发成 水产养殖水质净化高效微生态制剂。     

添加不同益生菌对草鱼养殖水体菌群结构的影响

为评价添加不同益生菌对草鱼养殖水体菌群结构的影响,研究采用454焦磷酸测序技术分析其水体菌群结构。结果表明:添加益生菌后的处理组（枯草芽孢杆菌BS、光合细菌PSB和复合菌CB）其微生物多样性高于对照组（Control）。在门的水平,Control和CB样品中变形菌（Proteobacteria）为优势菌,PSB和BS中变形菌（Proteobacteria）和放线菌（Actinobacteria）所占比例差别不大。与Control相比,其他三组中拟杆菌（Bacteroidetes）和放线菌（Actinobacteria）都增加。对变形菌深入分析发现,在PSB,BS和 CB 样品中,-变形杆菌为优势菌,接下来是-变形杆菌纲、-变形杆菌纲和-变形杆菌纲。对拟杆菌分析发现,除对照外,其他样品中黄杆菌纲（Flavobacteria）为优势菌。在对照和处理组中,-变形杆菌、-变形杆菌、-变形杆菌和拟杆菌门在目的水平组成也有差异。以上结果表明,水体中添加益生菌能增加水体菌群多样性,改变菌群结构。       

高效净水异养硝化细菌的筛选鉴定及固定化应用研究

本研究从海参养殖水体、泥土中筛选出4株具有硝化能力的异养硝化细菌。分别将其游离菌体细胞投入海参养殖水体,测定亚硝态氮、氨氮去除率,筛选出HS．NOB2为高效净化菌株,对HS-NOB2进行16SrDNA扩增及序列测定,初步鉴定为节杆菌（Arthrobactersp．）。利用海藻酸钠包埋法对高效净化菌体细胞进行固定化,将该固定化菌投入养殖水体及人工合成污水,研究其对水体中亚硝态氮、氨氮的处理效果,并与游离菌体细胞进行比较。结果表明,固定化后亚硝态氮去除率达到49．85％,氨氮去除率达到56．58％,均明显高于游离菌体细胞。上述研究为探寻水体净化提供了新思路,为水质改良剂的实际生产提供可选菌株。     

地衣芽胞杆菌对实验性家兔阴道炎影响的研究

目的通过探讨地衣芽胞杆菌活菌制剂对实验性家兔细菌性阴道病的影响,恢复家兔阴道微生态平衡和正常菌群环境。方法（1）采用注射用氨苄西林和甲硝唑生理盐水溶液注入家兔阴道进行冲洗,建立家兔阴道脱污染动物模型。（2）取40只阴道脱污染家兔,其中20只接种大肠埃希菌,20只接种金黄色葡萄球菌,建立家兔阴道感染模型。（3）地衣芽胞杆菌对家兔阴道菌群失调的调整作用：采用不同浓度的地衣芽胞杆菌菌液（10^6CFU／ml、10^8CFU／ml、10^9CFU／ml）对感染家免阴道进行接种,分析和考察地衣芽胞杆菌对家兔阴道菌群失调的影响以及对家兔阴道黏膜的影响。结果（1）动物经过金黄色葡萄球菌感染,通过地衣芽胞杆菌治疗后,动物阴道内芽胞杆菌、肠杆菌、乳杆菌数量明显上升,葡萄球菌数量明显下降,自细胞数量减少,黏膜红肿减轻、分泌物减少,治疗作用明显。（2）大肠埃希菌感染动物经地衣芽胞杆菌治疗后,动物阴道内芽胞杆菌、乳杆菌数量明显上升,肠杆菌数量明显降低,白细胞数量减少,黏膜红肿消失、分泌物减少。结论地衣芽胞杆菌对实验性家兔金黄色葡萄球菌和大肠埃希菌阴道炎的治疗有效。地衣芽胞杆菌与乳杆菌的作用相似,具有维持阴道菌群平衡的作用。     

蔗糖输入对凡纳滨对虾养殖系统真核微生物群落的影响

利用18S rRNA基因高通量测序, 研究蔗糖输入条件下凡纳滨对虾(Litopenaeus vannamei)养殖系统中水体与絮团的真核微生物群落组成及其相互作用关系。结果表明, 输入蔗糖能显著改变对虾养殖系统中真核微生物群落组成, 促进水体中纤毛虫和轮虫的生长, 对水体绿藻的过度生长有一定抑制作用。Mantel分析发现, 在第28天时, 蔗糖添加以及系统中的溶解氧、亚硝酸盐和磷酸盐对真核微生物群落结构变异有显著影响。利用共线性网络分析表明碳氮比的增加可以增强水体和絮团中不等鞭毛类、纤毛虫和轮虫的相互作用, 对养殖体系中藻类的过度繁殖具有一定的抑制作用。研究揭示了在蔗糖输入条件下凡纳滨对虾养殖系统中真核微生物群落的演替规律及絮团和水体微生物相互作用特征, 为生物絮团技术持续稳定的应用于对虾养殖系统提供理论支持。     

Massive production of all-female diploids and triploids in the crucian carp

In many species of aquaculture importance, all-female and sterile populations possess superior productivity due to faster growth and a relatively homogenous size of individuals. However, the production of all-female and sterile fish in a large scale for aquaculture is a challenge in practice, because treatments necessary for gynogenesis induction usually cause massive embryonic and larval mortality, and the number of induced gynogens is too small for their direct use in aquaculture. Here we report the massive production of all-female triploid crucian carp by combining artificial gynogenesis, sex reversal and diploid-tetraploid hybridization. Previously, we have obtained an allotetraploid carp population (4n = 200) by hybridization between red crucian carp (Carassius auratus red var; ♀) and common carp (Cyprinus carpio; ♂). We induced all-female diploid gynogens of the Japanese crucian carp (Carassius cuvieri; 2n = 100). We also generated male diploid gynogens of the same species treated gynogenetic fry with 17-α-methyltestosterone, leading to the production of sex-revered gynogenetic males. Finally, these males were used to cross with the female diploid Japanese crucian carp gynogens and the allotetraploid females, resulting in the production of fertile all-female diploid Japanese crucian carp (2n=100) and sterile all-female triploid hybrids (3n = 150), respectively. Therefore, diploid crucian carp gynogenetic females and sex-reversed male together with an allotetraploid line provide an opportunity to produce all-female triploid populations in a large scale to meet demands in aquaculture industry.     

Evidence for maternal inheritance of mitochondrial DNA in allotetraploid.

The complete mitochondrial DNA (mtDNA) sequences of the allotetraploid and red crucian carp were determined in this paper. We compared the complete mtDNA sequences between the allotetraploid and its female parent red crucian carp, and between the allotetraploid and its male parent common carp. The results indicated that the complete mtDNA nucleotide identity (99.7%) between the allotetraploid and its female parent red crucian carp was higher than that (89.0%) between the allotetraploid and its male parent common carp. Moreover, the analysis on the start and stop codons, overlaps and spacers, and phylogeny of the mt genomes indicated the genetic relationship between the allotetraploid and its female parent red crucian carp was closer than that between the allotetraploid and its male parent common carp. Our results indicated that the allotetraploid mt genome was strictly maternally inherited. Through maternal inheritance, the mt genome in the F(11) allotetraploid displayed extremely high similarity to that in the female parent red crucian carp after 11 generations (from F(1) to F(11) hybrids). Such results indicated that the F(11) allotetraploid possessed the stable inheritance characteristic. Thus the tetraploid stocks possessed the good base to form a new tetraploid species in the future. Since the establishment of the new tetraploid stocks has the great significance in analyzing evolutionary theory of vertebrate and in improving aquaculture industry, analysis of the mt genome and the elucidation of the variation of the mt genome in the allotetraploid and its parents proved that it was a useful genetic marker to monitor the variations in the progeny of the crosses.     

Performance of a constructed wetland in treating brackish wastewater from commercial recirculating and super-intensive shrimp growout systems

A recirculating aquaculture system was developed for treating Pacific white shrimp (Litopenaeus vannamei) production wastewater using an integrated vertical-flow (IVF) and five connected integrated horizontal flow (IHF) constructed wetlands as water treatment filters for mesohaline conditions (8.25‰-8.26‰ salinity). The constructed wetlands demonstrated the ability to reduce total nitrogen, total ammonia nitrogen, nitrite nitrogen, nitrate nitrogen, total phosphorous, chemical oxygen demand, and total suspended solids to levels significantly lower than those in effluents from culture tanks. Various water quality parameters in the culture tanks were deemed suitable for shrimp culture. The actual ratio of wetland area (A(w)) to culture tank area (A(t)) was 1.1439, and the estimated optimal ratio A(w)/A(t) was approximately 1. The IVF-IHF wetlands showed flexibility and reliability in consistently removing the main pollutants from commercial recirculating and super-intensive shrimp growout systems throughout the culture period.     

Comparative and Evolutionary Analysis of the Cytochrome b Sequences in Cyprinids with Different Ploidy Levels Derived from Crosses

The mitochondrial cyt b genes in the allotetraploid and triploid crucian carp as well as triploid common carp were isolated and completely sequenced. Their DNA sequences were compared with those derived from the cyt b genes of the red crucian carp, Japanese crucian carp, and common carp with MEGA 1.0 software. Phylogenetic analysis revealed the sister relationships between allotetraploid and diploid red crucian carp, between the triploid crucian carp and diploid Japanese crucian carp, and between triploid common carp and diploid common carp. Our results indicated the cyt b genes in the allotetraploid, triploid crucian carp, and triploid common carp were maternally inherited. Through maternal inheritance, the cyt b gene in the F11 tetraploid displayed extremely high similarity to that in the female parent red crucian carp after 11 generations (from F1 to F11 hybrids). Since the establishment of the new tetraploid stocks has great significance in analyzing evolutionary theory of vertebrate and in improving aquaculture industry, analysis of the cyt b gene and the elucidation of the variation of the cyt b gene DNA in different cyprinids prove that cyt b is a useful genetic marker to monitor the variations in the progeny of the crosses.     

一株枯草芽孢杆菌的分离鉴定、生物学特性及其对水质净化的作用

[背景]随着水产养殖业的发展和养殖集约化程度的提高,养殖水环境日趋恶化,养殖动物病害频发,而水产益生菌因其环境友好、安全而被广泛应用于水产养殖中.[目的]从南美白对虾养殖池底泥分离枯草芽孢杆菌,探究其体外生物学特性及对水质的净化作用,以期扩充微生态制剂的种质资源.[方法]采用稀释涂布平板法分离菌株,通过形态学观察、生理...     

氨氮和镉胁迫对芙蓉鲤鲫抗氧化系统和免疫机能的影响

为探究氨氮、镉单一及联合胁迫对鱼类氧化防御系统及非特异性免疫系统的影响, 以芙蓉鲤鲫(Cyprinus capio furong.♀×Carassius auratus red var.♂)为研究对象, 通过静态毒理学实验方法模拟亚急性胁迫, 在胁迫后不同时间点对芙蓉鲤鲫的肝脏进行取样, 并测定其抗氧化指标和非特异性免疫指标。结果显示, 氨氮和镉单一及联合胁迫后肝脏丙二醛(MDA)含量、酸性磷酸酶(ACP)活性和髓过氧化物酶(MPO)活性均出现不同程度的升高, 过氧化氢酶(CAT)活性均呈先下降后升高的趋势。氨氮胁迫后肝脏超氧化物歧化酶(SOD)活性、谷胱甘肽过氧化物酶(GSH-PX)活性、碱性磷酸酶(AKP)活性及谷胱甘肽(GSH)含量均呈先升高后下降的变化趋势。镉胁迫后肝脏SOD和GSH-PX活性在6d时均显著高于对照组, GSH含量在4d时显著低于对照组, AKP活性呈先降低后升高的趋势。氨氮和镉联合胁迫后肝脏SOD和GSH-PX活性在6d和8d时显著增加, AKP活性在8d时显著升高, GSH含量呈先降低后升高的变化趋势。结果表明, 氨氮和镉单一及联合胁迫均可诱导芙蓉鲤鲫产生氧化应激, 激活了鱼体的氧化防御系统和非特异性免疫系统。研究可为深入探讨芙蓉鲤鲫的健康养殖和逆境生理响应机制提供基础资料。     

Closed water recirculating system for fish rearing equipped with bioreactor capable of simultaneous nitrification and denitrification.

Five crucian carp, Carassius auratus langsdorfiicarps had been reared in a closed water recirculating system. The system was equipped with the compact bioreactor using the plate gels capable of both nitrification and denitrification in a single unit. Ammonia and nitrite concentrations in the rearing water had been maintained below 0.05 mg-N/L, and nitrate concentration also controlled between 2 and 8 mg-N/L with the bioreactor. As concerns nitrogen budget in the closed system, 95.0% of nitrogen income from feed was lost as nitrogen gas from the closed system. All fish was alive for 91 days without any unusual behavior. Thus, the bioreactor performed both nitrification and denitrification abilities enough to rear the five fish for 91 days. The bioreactor using the plate gels would be effective to simplify the closed system both physically and operationally, since it can remove the ammonia excreted from fish as nitrogen gas by a single step.     

海洋养殖环境微生物多样性及其作用

微生物是海洋养殖环境和海洋牧场生态系统中的关键环境因素之一，在牧场环境的物质循环、水质保障、饵料供应、水产健康等方面起重要作用，是营造海洋牧场环境且影响其可持续发展的重要因素。主要围绕海洋养殖环境中的病原微生物以及有益微生物两方面，概述了我国海洋养殖环境常见的病原性细菌、真菌、病毒以及相应的病害防治措施等，并介绍了海洋养殖环境中能够改善海水养殖环境，包括降低水体氨氮含量、降解水体硫化物、抑制赤潮藻类的生长等以及直接促进海水养殖生物生长的有益功能微生物类群。目前，我国关于海洋牧场环境营造过程中微生物的研究仍很少，而与海洋牧场环境有一定相似性的海水养殖环境中微生物的研究较为成熟，其对海洋牧场环境健康营造有很好的启示。海洋水产养殖环境中微生物的新功能菌种的发现、作用机制以及功能菌应用等方面的深入研究将会对我国大力推进海洋牧场建设起到重要的作用。