淡水入侵生物沼蛤的污损机制与防污措施研究进展

沼蛤是一种典型的淡水入侵贝类,能够利用其分泌的足丝牢固黏附在多种水下基质表面,引起严重的生物污损问题。沼蛤污损不但影响水生态系统健康,也给水利工程、交通运输、水产养殖等行业带来经济损失,已成为全球水生态系统安全和国民经济重要行业的潜在威胁,相关防污工作亟待开展。欲从根本上解决沼蛤污损问题,一方面需要加强对其基础生物学特性和污损机制的深入解析,另一方面也需要在此基础上研发更加经济、高效、环境友好的防污措施。本文综述了近年来国内外关于沼蛤污损生物学特性、污损机制和防污措施方面的研究进展,尤其是对沼蛤生物污损发生的主要机制如足探测识别、足丝黏附和环境影响等方面进行了总结,也从物理、化学、生物和防污材料等角度阐述了现有的沼蛤污损控制措施并对未来发展方向进行了展望,以期更加深入地理解沼蛤生物污损现象,为揭示其作用机制、制定科学有效的防污措施、维护水生态系统安全提供数据支撑,综述内容对于水下仿生材料研发也具有重要的参考价值。     

凌云沼虾(Macrobrachium lingyunense)

凌云沼虾(Macrobrachium lingyunense u,Cai & Clarke 2006)(封面图片)是一种典型的洞穴沼虾.眼睛极度退化,步足等外部形态特征及其他生物学特性也显示出真洞穴生物(troglobiont)的特征,包括缺乏体色素、附肢延长、繁殖无季节性、耗氧量降低、新陈代谢缓慢等.     

Notch信号途径中的Su(H)和E(spl)基因对果蝇天然免疫的影响

天然免疫系统是多细胞生物抵抗各种入侵微生物的第一道防线.Notch途径介导相邻细胞之间的相互作用,调节细胞、组织、器官的分化和发育.为了进一步探索Notch信号途径在果蝇天然免疫中的功能,利用Notch途径下游基因Su(H)和E(spl)的低表达突变体果蝇,通过体外注射病原体分析了生存率、血细胞的噬菌功能和抗菌肽的表达量以及突变体的血细胞数量.结果表明,革兰氏阴性细菌和真菌感染后果蝇E(spl)突变体的生存率、噬菌能力及抗菌肽的表达量明显降低,而且幼虫期血细胞出现异常增殖;Su(H)突变体只对真菌表现出敏感性,抗菌肽的表达量降低,但是对真菌的噬菌能力正常.此结果表明,Notch途径不仅影响个体的生长发育,而且在果蝇天然免疫中也起重要的调节作用.     

大豆品种(系)和种植密度对豆天蛾幼虫存活及生长发育的影响

豆天蛾Clanis bilineata tsingtauica Mell幼虫,是一种食用昆虫,富含蛋白质、维生素和矿质元素,具有很高的营养、药用和经济价值。了解适宜豆天蛾幼虫生长发育的大豆品种（品系）和种植密度,可为豆天蛾幼虫规模化饲养提供理论基础。研究通过设置两因素裂区实验,在10个品种（品系）大豆和2个种植密度水平下,测定豆天蛾幼虫生物学参数,明确适宜豆天蛾幼虫养殖的大豆品种（品系）和种植密度。结果表明,不同大豆品种（系）和种植密度对豆天蛾幼虫存活及生长发育均有显著影响;其中使用连151、淮豆9号、东辛3号和H0573大豆饲养的豆天蛾幼虫产量和存活率较高,虫体长度较长,发育历期较短;大青豆饲养的豆天蛾幼虫产量较低,幼虫发育整齐度较差,发育历期长。当种植密度为40 cm×30 cm时,豆天蛾幼虫存活和发育历期整齐度均显著高于种植密度40 cm×15 cm。综上所述,较适宜豆天蛾幼虫养殖的大豆品种（品系）为连151、淮豆9号、东辛3号和H0573,种植密度为40 cm×30 cm。     

絮凝酵母吸附Cr(VI) 的机理

絮凝酵母SPSC01为酿酒酵母Saccharomyces cerevisiae和粟酒裂殖酵母Schizosaccharomyces pombe的融合菌株,用其吸附水溶液中的重金属Cr(VI),可以大大降低生物吸附的固液分离成本。为了探讨SPSC01菌体絮凝蛋白对Cr(VI) 还原吸附的影响,对SPSC01与其亲本菌株的吸附行为进行了比较。结果表明,SPSC01和其具有絮凝性状的亲本S. pombe的Cr(VI) 去除速率基本同步,远优于无絮凝性状的亲本S. cerevisiae;达到吸附平衡时,S. pombe、SPSC01和S. cerevisiae对总Cr去除率分别达68.8％、48.6％和37.5％;从而证明了絮凝有利于Cr(VI) 的还原、吸附,絮凝蛋白在Cr(VI) 的还原吸附过程中起促进作用。通过化学屏蔽方法和傅立叶变换红外光谱 (FTIR) 分析,对SPSC01菌体表面吸附Cr(VI) 的机理进行了研究,结果表明SPSC01菌体表面吸附Cr(VI) 起主要作用的基团是氨基、羧基和酰胺基。     

中华蜜蜂囊状幼虫病病毒单克隆抗体的制备与鉴定

为了制备中华蜜蜂囊状幼虫病病毒(CSBV)单克隆抗体,本实验利用纯化的中华蜜蜂囊状幼虫病毒(CSBV)免疫Balb/c小鼠,经过3次免疫后,小鼠断尾采血测其血清效价,选择效价高于1∶80 000的小鼠,在细胞融合前3~4d再加强免疫一次。取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合,用间接ELISA筛选阳性细胞株并进行3次亚克隆后注射入Balb/c小鼠(Mus musculus)腹腔制备单抗腹水。共获得9株稳定分泌抗CSBV的单克隆抗体,命名为10A1、5B10、5A5、5H2、11D7、9A5、1D3、3C10、10C4,效价都在1∶16 000以上,经间接ELISA实验检测表明,具有较好的特异性。在此基础上,将9株腹水单抗分别与CSBV互作后,接种于2~3日龄中华蜜蜂幼虫,观察幼虫死亡率,研究单克隆抗体中和CSBV病毒能力,结果表明筛选到三株单克隆抗体(10A1、5A5、9A5)对CSBV有中和作用,为CSBV的防治研究奠定了基础。     

缅甸蛤(Burmesia)在四川西北部的发现

1957年,某地质队在四川西北部江油一带的黑色页岩钻心中,发现了一些瓣鳃类化石,寄送我所请顾知微教授鉴定。最近,顾教授将上述标本交由作者研究,鉴定结果,包括有BurmesialirataHealey及Posidoniasp.indet.(sp.nov.?)两个种。上述化石的地面地层层位尚不清楚,现仅将某队在江油官渡地区所作的野外地层剖面列在下面,以供参考。     

絮凝酵母吸附Cr(VI)的机理

絮凝酵母SPSC01为酿酒酵母Saccharomyces cerevisiae和粟酒裂殖酵母Schizosaccharomyces pombe的融合菌株,用其吸附水溶液中的重金属Cr(VI),可以大大降低生物吸附的固液分离成本。为了探讨SPSC01菌体絮凝蛋白对Cr(VI)还原吸附的影响,对SPSC01与其亲本菌株的吸附行为进行了比较。结果表明,SPSC01和其具有絮凝性状的亲本S.pombe的Cr(VI)去除速率基本同步,远优于无絮凝性状的亲本S.cerevisiae;达到吸附平衡时,S.pombe、SPSC01和S.cerevisiae对总Cr去除率分别达68.8%、48.6%和37.5%;从而证明了絮凝有利于Cr(VI)的还原、吸附,絮凝蛋白在Cr(VI)的还原吸附过程中起促进作用。通过化学屏蔽方法和傅立叶变换红外光谱(FTIR)分析,对SPSC01菌体表面吸附Cr(VI)的机理进行了研究,结果表明SPSC01菌体表面吸附Cr(VI)起主要作用的基团是氨基、羧基和酰胺基。     

两种沼虾溶菌酶基因ORF的克隆和罗氏沼虾溶菌酶基因的组织表达(英文)

分别提取罗氏沼虾和日本沼虾血细胞总RNA,RT-PCR扩增获得特异性cDNA片段,纯化后克隆到T载体上。序列测定表明所克隆的两种沼虾溶菌酶基因的开放阅读框(ORF)为477bp,共编码158个氨基酸,包括溶菌酶成熟肽140个氨基酸残基和信号肽18个氨基酸残基。同源性分析表明,罗氏沼虾和日本沼虾溶菌酶基因的碱基序列及推测氨基酸序列高度同源,分别为99.4%和98.1%。两种沼虾溶菌酶基因的碱基序列和推测氨基酸序列与Gen-Bank上其他对虾溶菌酶的同源性达83.0%和80.0%以上。两种沼虾溶菌酶都具有c-型溶菌酶典型的两个酶活性位点(Glu51)和(Asp68),以及8个保守结构氨基酸残基Cys,且在101、106和107位上缺少Asp,因而推测本实验所克隆的两种沼虾溶菌酶基因属c-型溶菌酶基因的非钙结合亚型。以PCR法制备罗氏沼虾溶菌酶基因的生物素标记探针,斑点杂交检测感染弧菌后溶菌酶基因mRNA在各组织中的转录水平,结果表明受感染6h后在眼、肌肉、鳃、肝胰腺、肠管中的表达量均有升高,其中在肝胰腺中的表达量最高,约为对照组的560%。在不同感染时间里,肝胰腺中该基因表达量有较大的变化:感染后3h表达量最低,24h后表达量升至最高,大约为对照组的430%,48h时的表达量又有所下降,但仍明显高于对照组(约为330%)。受弧菌感染后罗氏沼虾溶菌酶基因转录的上调证明溶菌酶基因在非特异性免疫中的直接作用,同时表明肝胰腺可能在沼虾的免疫防御过程起重要作用。       

秦岭大坪峪川金丝猴(Rhinopithecus roxellana)种群生存力分析

小种群的保护已成为保护生物学中一个重要的问题,鉴于小种群极易灭绝的特性,寻找制约小种群存活的关键因子,形成快速拯救的措施,是目前对于小种群进行保护十分迫切的任务。根据秦岭大坪峪地区2个川金丝猴(Rhinopithecus roxellana)种群连续5a的观察数据,参照群体大小、配偶体制、迁移扩散、环境容纳量、死亡率、灾害的发生频率等种群参数,并结合川金丝猴其他种群的研究结果,利用旋涡模型(Vortex 10.0),对猴群未来100a内的数量动态进行了模拟分析。结果表明,在没有个体迁移的情况下,大坪峪种群种群100a间灭绝概率为95.8%和93%;当存在个体迁移和扩散的情况,灭绝概率为2%和0.4%。因此种群间个体迁移是秦岭大坪峪川金丝猴种群数量动态的限制因子。鉴于此,增加川金丝猴大坪峪种群间的个体交流,能够保证本群川金丝猴长期生存所需的遗传多样性。同时,增加秦岭川金丝猴各种群间的个体交流,建立群间的生境走廊对这一世界性濒危物种的长期存活具有重要的现实意义。     

环境DNA技术在贻贝入侵监测中的应用挑战

沼蛤(Limnoperna fortunei)和斑马贻贝(Dreissena polymorpha)是淡水系统中常见的入侵贻贝物种,对其种群规模的持续监测是入侵贻贝防治管控中的关键环节。随着分子生物学技术的发展,入侵物种监测中逐渐尝试利用环境DNA(eDNA)技术实现快速、灵敏检测。然而,在入侵物种引入-定植-扩散过程的监测中,eDNA技术的灵敏度及定量效果受到诸多因素的影响,给实际应用带来挑战。系统梳理了国内外学者利用eDNA技术监测沼蛤、斑马贻贝等入侵物种的研究进展;分析了eDNA技术的采样方案、引物设计、定量分析、质量保证、原位便携仪器设计等影响监测效率与准确率的关键环节;进一步探讨了eDNA技术在贻贝入侵监测中的优势和局限性,以及未来的改进方向。     

Invasion by Limnoperna fortunei (Dunker, 1857) (Bivalvia, Mytilidae) of the Pantanal Wetland, Brazil

Limnoperna fortunei (Bivalvia, Mitylidae) was introduced into South America in 1991 in the La Plata River (Argentina). It arrived in the ballast water of ships coming from Asia, where this species is native. It was first observed in 1998 in the Paraguay River. Limnoperna was introduced into the Pantanal region as hull fouling of vessels using the Paraguay–Parana waterway. This study describes how L. fortunei came to the Pantanal region, and provides details of its occurrence, density, and impacts. From 1999 to 2002, observations and sampling on natural and artificial substrates in the Paraguay River were made. Some aspects of the spread and impacts, based on local community information, were also analyzed. On artificial substrate the density reached 523.8 individuals m⁻² and on natural substrate (rocks), up to 10,000 individuals m⁻² were found. The densities observed were quite low compared to those found in Southern Brazil, where values up to 100,000 individuals m⁻² have been recorded in the last 3 years. In the Paraguay River, the population density of L. fortunei can be negatively impacted by periodic low levels of dissolved oxygen and decreases in pH to between 5 and 6. Such conditions are frequently present during the periodic flooding or inundation of this area. Under these conditions, a high mortality of L. fortunei was recorded in March of 2002, on both natural and artificial substrates. Despite low densities, L. fortunei can colonize water cooling systems of boats, obstructing water circulation and causing motor overheating. Accumulation in water supply equipment, such as pumps and pipes has also been observed. An erratum to this article is available at .     

北京典型水域贝类和虾类生物多样性监测及入侵风险分析

【目的】了解北京地区典型水域贝类和虾类生物多样性,分析外来水生入侵物种的生态风险。【方法】选取北运河、潮白河、永定河、大清河和蓟运河五大水系以及南水北调蓄水工程沿线等北京地区典型水域,于2020年6、8和10月开展3次监测工作。共设置监测站点48个,涵盖河流、湖泊、水库等不同水体类型。通过对采集样品的分类鉴定,分析北京地区贝类和虾类物种组成、分布特征和生物多样性,同时对入侵贝类沼蛤的生态风险进行初步分析。【结果】本研究共鉴定出贝类和虾类13科27种,其中,贝类11科22种,虾类2科5种。角形环棱螺、纹沼螺、中华新米虾和铜锈环棱螺为各监测站点中的优势物种。从监测时间分析,各监测站点贝类和虾类的物种数量、密度和生物量都随着监测时间的延续表现为先升高后降低的趋势,而生物多样性表现为持续升高趋势。从水体类型分析,各监测站点贝类和虾类的物种组成和生物多样性没有表现出显著差异,同质化趋势明显,而湖泊型水体贝类和虾类的生物量和密度较高。此外,本研究在玉渊潭西湖和龙潭闸2个站点均采集获得入侵贝类沼蛤的活体样品,且在3次监测中沼蛤种群密度均较大,这表明北京地区沼蛤生物入侵风险较大,需持续开展跟踪监测并密切关注其种群发展动态。【结论】北京地区底栖动物尤其是贝类和虾类资源较为丰富,但相关监测调查仍然没有全面覆盖本地贝类和虾类物种,在后续工作中应该加大监测力度,同时应密切关注入侵生物沼蛤的种群动态和入侵情况。     

Detachment,displacement and reattachment activity in a freshwater byssate mussel (Limnoperna fortunei): the effects of light,temperature and substratum orientation

The ability of the freshwater bivalve Limnoperna fortunei to voluntarily detach from the substratum, crawl and reattach as a function of illumination, temperature, substratum orientation, and mussel size was investigated. Thirty-two per cent of the 879 experimental animals detached and reattached elsewhere at least once during five- to eight-day experiments. The proportions of mobile mussels were significantly higher in permanent darkness than under permanent illumination. Displacement distances were also higher in darkness, but statistical differences with illuminated individuals were inconclusive. No evidence of circadian rhythms was detected. Mobile mussels were often significantly smaller than non-mobile individuals. It was not possible to detect the effect of water temperature (22°C and 31°C), or substratum orientation (topside and underside) on mussel mobility, but because the power of the statistical tests was low, future experiments are needed to confirm this result. The ability of mussels to voluntarily detach and reattach elsewhere has important implications for biofouling control.     

Attachment strength of an adhesive nuisance mussel,limnoperna fortunei,against water flow

The attachment strength of the freshwater mussel Limnoperna fortunei against water flow was studied. Newton's expression successfully described the hydrodynamic drag force acting on the mussel with a drag coefficient value of 1.03. The drag‐resistant force (defined as hydrodynamic drag force at mussel detachment) was smaller than the detachment force measured using a tensile load test. A fairly good correlation was obtained between the drag‐resistant force and the number of secreted threads. The drag‐resistant force divided by the number of threads increased with shell size, suggesting that byssal thread strength increased with mussel growth. For the mussel specimens obtained from a water transmission pipe, thread width increased with shell size. However, thread width was not dependent on current velocity. There was no correlation between the number of secreted threads and shell length, which indicated that the number of secreted threads did not change with mussel size. Therefore, the water velocity needed to detach mussels increases with shell size of the mussel when the number of secreted threads is constant. The increases in the water velocity to detach mussels with larger shells suggests that the mussel becomes more resistant to water flow as it grows. It is estimated that a flow velocity of around lms^‐1 is critical for attachment/detachment of a juvenile mussel with a shell length of a few millimeters and one hundred byssal threads.     

Effects of different biotic substrata on mussel attachment

Surface colonization by invertebrates can be stimulated or inhibited by cues produced by biofilms, conspecifics or other macroorganisms. To study the effects of living substrata on the attachment of the brown mussel, Perna perna, two different approaches were employed: (1) mussels were distributed in sets of Petri dishes consisting of one sterile set (controls), three sets in which marine biofilms were allowed to develop in aquaria for 1, 7 or 15 days and another set that had been immersed in a natural marine environment for 1-day. There was no significant effect of biofilms on attachment, suggesting that neither age nor the source of the biofilm influenced attachment. (2) Mussels were suspended over PVC panels (controls) and over panels on which Balanus trigonus (Crustacea), Schizoporella errata (Bryozoa), Symplegma rubra or Didemnum speciosum (Ascidiacea) were present. Attachment was significantly higher on the controls and on B. trigonus than on colonial taxa such as S. rubra, S. errata and D. speciosum, probably due to antifouling defenses of these species. The results show that the composition of the biological substratum is an important factor affecting mussel behavior.     

Combined effects of food quantity and quality on Cd, Cr, and Zn assimilation to the green mussel, Perna viridis

We examined the assimilation of Cd, Cr, and Zn by the green mussel Perna viridis under complicated food conditions, including combinations of different compositions and concentrations of food (diatom and sediment), and variable food quantity and quality during particle digestion. At different combinations of food composition and quantity (5 mg l⁻¹ and 20 mg⁻¹, below and above the pseudofeces production), the Cd assimilation was significantly dependent on the food composition. The Cd assimilation efficiency (AE) decreased with increasing proportions of sediments in the diets, but its assimilation was not significantly affected by food concentration. In contrast, the assimilation of Cr and Zn decreased significantly with increasing food concentrations, whereas food composition did not significantly affect their AEs. Variations in metal gut passage time accounted partially for the difference in AEs among different combinations of food composition and quantity. By changing the type of particles during metal digestion, their AEs were maintained comparably at a low particle load (1 mg l⁻¹), suggesting that variation of food quality during digestion did not affect metal assimilation. At a higher particle load (5 mg l⁻¹), variation of food type during digestion affected the AEs of Cr and Zn. An increase in food concentrations from 1 to 15 mg l⁻¹ during digestion resulted in a significant decrease in the AEs of Cr and Zn bound with either sediments or diatoms. Conversely, decreasing the food concentrations from 15 to 1 mg l⁻¹ did not affect the AEs of metals, except for Zn bound with diatoms. Overall, our results highlighted the metal-specificity in their assimilation as influenced by complicated food environments, probably caused by different metal geochemical and biological behavior in the mussels. Feeding selectivity may have a greater control on the influx rate into the mussels than metal assimilation.     

The role of bacterial attachment in the transformation of cell-wall-regenerating tobacco protoplasts by Agrobacterium tumefaciens

The presence of a newly formed primary cell wall was shown to be required for attachment and subsequent transformation of tobacco leaf protoplasts by Agrobacterium tumefaciens in cocultivation experiments. In these experiments both protoplasts at different stages after their isolation and cell-wall inhibitors were used. The specificity of Agrobacterium attachment was shown by using other kinds of bacteria that did not attach. By diminishing the concentration of divalent cations using ethylenediaminetetraacetic acid, neither attachment nor transformation was found; however, when more specifically the Ca²⁺concentration was lowered by ethylene glycol-bis (-aminoethyl ether)-N,N,N,N-tetraacetic acid, both phenomena occurred. Commercial lectins had no effect on binding, but this observation does not exclude the involvement of other lectins. Protoplasts isolated from various crown-gall callus tissues also developed binding sites, but when they were at the stage of dividing cells, attachment of agrobacteria was no longer observed. In this respect, cells from protoplasts of normal tobacco leaves behaved differently. Even 16 d after protoplast isolation, the dividing cells were still able to bind A. tumefaciens, while transformation was not detected. For transformation of 3-d-old tobacco protoplasts, a minimal co-cultivation period of 24 h was required, while optimal attachment took place within 5 h. It is concluded that the primary cell wall was sufficiently well formed that certain functional receptor molecules were available for attachment of Agrobacterium as the first step of a multistep process leading to the transformation of cells. The expression of bacterial functions required for attachment, moreover, was independent of the presence of Ti-plasmid.Abbreviations ConA concanavalin A - CW calcofluor white - EDTA ethylenediaminetetraacetic acid - EGTA ethylene glycol-bis (-aminoethyl ether)-N,N,N,N-tetraacetic acid - -Man -methyl-d-mannoside     

Studies on fouling by the freshwater mussel limnoperna fortunei and the antifouling effects of low energy surfaces

Attachment of the freshwater mussel, Limnoperna fortunei, was tested using non‐treated surfaces, viz. glass, nylon, rubber, silicone and Teflon, together with glass surfaces modified with nine kinds of silane coupling agents. Among the surfaces tested, the mussel avoided attaching to Teflon, silicone, and glass modified with 3‐bromopropyltrimethoxysilane or 3,3,3‐(trifluo‐ropropyl)‐trimethoxysilane. With respect to the relationship between the percentage attachment and the surface free energy (sfe) of the substrates, it was found that attachment was considerably reduced on the substrates which exhibited relatively low sfe, as above. The mean number of secreted byssuses per attaching mussel also decreased with decreasing substrate sfe. Furthermore, when the sfe was divided into the dispersion and polar components, the percentage mussel attachment was related to the polar component. These results suggest that effective antifouling towards L. fortunei is achieved on substrates with a low sfe polar component.