深海嗜冷希瓦氏菌Shewanella psychrophila WP2的基因组学分析

嗜冷希瓦氏菌Shewanella psychrophila WP2分离自深海沉积物,其拥有目前希瓦氏菌属(Shewanella)中最大的基因组,为6. 4 Mb。通过对S. psychrophila WP2的完整基因组进行系统发育和代谢潜能分析,并与相近菌株进行比较,探究WP2拥有较大基因组的生理和生态意义,深化对Shewanella菌在环境适应性方面的认识。基于串联保守蛋白对WP2基因组进行系统发育分析,根据注释结果对WP2进行代谢通路重建。系统发育进化树显示,WP2属于Group 1分支,和其它嗜冷嗜压Shewanella菌相同,非嗜冷嗜压Shewanella菌株都属于Group 2分支。WP2基因组中与DNA修复、次级代谢产物合成、转运分泌相关的基因数明显多于Shewanella piezotolerans WP3。WP2拥有较多水平转移来源的基因组岛(17个),岛内包含了与嘌呤代谢、氨基糖代谢、运动趋化、群体感应等功能相关的基因。结果表明,通过潜在的水平基因转移,WP2获得了大量的辅助功能基因,增强了它的氮源利用能力、运动和趋化能力,以及应对复杂环境的群体感应能力和抗生素合成功能,从而更加适应生活在寡营养和复杂极端的深海环境。本研究针对S. psychrophila WP2的代谢潜力进行了深入的探索和研究,为将来关于Shewanella菌和微生物的环境适应机制的研究提供了更多数据参考和理论基础。     

Biophysical models of larval dispersal in the Benguela Current ecosystem

We synthesise and update results from the suite of biophysical, larval-dispersal models developed in the Benguela Current ecosystem. Biophysical models of larval dispersal use outputs of physical hydrodynamic models as inputs to individual-based models in which biological processes acting during the larval life are included. In the Benguela, such models were first applied to simulate the dispersal of anchovy Engraulis encrasicolus and sardine Sardinops sagax ichthyoplankton, and more recently of the early life stages of chokka-squid Loligo reynaudii and Cape hakes Merluccius spp. We identify how the models have helped advance understanding of key processes for these species. We then discuss which aspects of the early life of marine species in the Benguela Current ecosystem are still not well understood and could benefit from new modelling studies.     

Symbiotic Relations of Sediment-Agglutinating Nematodes and Bacteria in Detrital Habitats: The Enzyme-Sharing Concept

Abstract. A new concept (‘enzyme sharing’) concerning the interaction of marine nematodes and microbes in the degradation of sedimentary detritus is presented. Elements of this concept are (1) the notorious tendency of many aquatic nematodes to agglutinate detrital particles by mucus secretions, (2) new observations on the stimulation of microbial growth by nematodes in agar plates, and (3) literature data on limited endogenous proteolytic capacities of aquatic nematodes.
Observations on nematode‐microbe associations in agar plates prompted the conceptual synthesis. In agar medium without the addition of any nutrients a spectacular growth of bacteria was visible on the sinusoidal crawling trails of nematodes only 2 – 3 days after introduction of the worms (species of Adoncholaimus, Anoplostoma and Sabatieria). Juveniles of Anoplostoma that hatched in the agar cultures left their minute trails in the medium and these were rapidly occupied by bacteria. The nematodes repeatedly visited their bacterial trails, which persisted as a peculiar biotic structure for more than one year and survived the nematodes.
In sterile agar preparations containing the fluorogenic methylumbelliferyl‐β‐glucoside in the presence of the nematode Adoncholaimus, an enhanced fluorescence of the medium was visible, indicating β‐glucosidase activity. We therefore assume that oncholaimid nematodes discharge enzymes that alone, or in concert with microbial activities, contribute to the hydrolytic cleavage of refractory polysaccharides containing β‐glucosidic bonds such as agar components and cellulose. The sugars thus produced may then be taken up by the nematodes and concomitantly support the conspicuous growth of microbes. Since we did not observe any feeding of the nematodes on the associated microbes in agar plates, we question the nutritive potential of intact microbial cells for a number of nematodes abounding in detrital habitats, and call attention to the significance of ambient dissolved or adsorbed organic monomeric nutrients.
Consequently, we perceive the puzzling perpetual accretion of detrital organic particles to sediment agglutinations by nematodes as an adaptation for operating an ‘enzymatic reactor’ for the production of dissolved nutrients. We hypothesise a relationship of mutual commensalism of nematodes and heterotrophic microbes in detrital habitats and propose the term ‘enzyme sharing’ for this relationship. Both parties invest in a common enzyme pool that decomposes organic detritus for their nutrition. We present here evidence that nematodes contribute β‐glucosidase, which is involved in the cellulase system. Data from the literature suggest that microbial enzymatic processing of detrital proteins yields amino acids available to nematodes, which apparently have no efficient proteolytic enzyme system in their intestines.     

海水养殖环境细菌耐药性的危害

为解决人口、资源、环境等人类面临的重大问题，海洋作为地球上尚未充分开发的重要疆域而备受重视。中国以海水养殖业为基础的“蓝色农业”在缓解人口增长对陆地农业的压力，促进沿海地区经济的发展中正发挥着重要作用。然而，近10年来，世界范围内海水微生物流行性和暴发性病害相继发生，严重制约了海水养殖业的健康发展(相建海，2003)。为了防治细菌性病害，大量的抗生素药类被应用于养殖过程的各个阶段，巨大的选择压力导致了海水养殖水域耐药细菌的种类和数量的显著增加，养殖生物耐药病原菌病害的发生也越来越频繁，微生物病害防治的难度越来越大。更严重的问题是这样培养的水产养殖品往往携带耐药细菌，通过食物链传递，将对人类健康和生命安全直接构成威胁(Sorum et al.，2002)。一些海产生物病原菌本身也是人类病原菌，如一些海洋弧菌(French et al.，1989；Wong et al.，2000)，这些水陆两栖人畜共患病原菌耐药性的获得及这种病害的发生和流行将会给人类带来灾难性的后果。另外，由于大多耐药因子的可传播性，海水养殖环境中正常菌群的耐药性同样是一个不可忽视的严峻问题(Sorum et al.，2002)。大多数抗性决定子(耐药基因)是在可移动和传递的遗传载体，如质粒或接合转座子上，可在不同菌株甚至是不同种属细菌间相互传递(Ferber，1998；OBrien，2002)。再加上养殖水体的半封闭性(池养中的定期或不定期的换水)或全开放性(滩涂或近海的网箱或筏式养殖)，耐药细菌有可能被潮汐和海流带入近岸的非养殖区及海滨旅游区，通过菌间遗传物质的传递，耐药基因有可能被传入环境中的人体病原菌，大大增加了人体耐药病原菌病害发生和流行的可能性及危害程度(Miranda et al.，2001；Thayumanavan et al.，2003)。大量抗生素的使用还改变了养殖海域及临近自然海区细菌群落的结构及种类和功能多样性，破坏了生态平衡和海洋正常的生命过程(Chelossi et al.，2003)。养殖水域作为可能的抗药因子的孳生地、储藏库和扩散源(Miranda et al.，2002；Rhodes et al.，2000)，已引起了全球性的关注。世界上一些水产养殖国家相继开展了养殖水体耐药细菌的调查研究，并有一些国家已对养殖中抗生素的种类、使用方法、使用剂量、残存量及检测标准进行了行政化的规范和管理(李爱华，2002)。目前我国对海水养殖水体细菌耐药危害的认识明显不足，许多需要进行的研究尚未开展，为更有效地防治耐药细菌病害，避免耐药基因在海洋环境中的广泛传播扩散，保护海洋生态环境及资源，提高养殖产品卫生质量，保护海滨旅游环境及人民群众身体健康，在我国开展海水养殖环境耐药细菌及其抗性机理的调查研究已是刻不容缓，这项工作的重点应在于查清耐药因子的源及其传播途径和动态，并在此基础上寻找治理的科学办法，以达到防患于未然的目的。20世纪滥采乱捕导致的近海渔业资源的严重衰竭是一个非常深刻的教训，希望这种急功近利、涸泽而渔的状况不会再度在海产养殖业中出现。我们应该把海洋细菌耐药性的研究提升到直接关系到人类生存和发展的环境和资源问题的高度来对待，避免海洋成为耐药因子孳生、繁衍、传播和扩散的潘多拉魔盒。     

法紫菜生物多样性及其栽培生物学基础

综述了重要经济红藻法紫菜属物种多样性及其地理分布,修订了法紫菜属中文名称,回顾了紫菜生活史研究的历程,基本概括了法紫菜栽培生物学基础研究成果。根据文献报道及最新研究成果,认为紫菜属和法紫菜属共有138个物种,集中分布在北半球的温带、亚热带海域,其中我国是物种多样性较高的国家之一,其物种及变种数达到25个。法紫菜生活史的阐明对紫菜栽培产业的发展具有奠基性的作用,其中我国藻类学家曾呈奎和张德瑞在法紫菜生活史研究中作出了重要的贡献,提出了壳孢子的概念,被广泛接受并沿用至今。壳孢子采苗和丝状体培养技术是发展紫菜栽培业的重要基础,丝状体生长发育及壳孢子形成的分子机制将是未来法紫菜生物学研究的重点和难点。尽管根据已发表的相关文献大致梳理了法紫菜相关生物学基础研究的成果,但限于篇幅对栽培生产中相关技术的优化、完善与推广以及新品种培育等均未作介绍。     

Chemical and Biological Studies of Particulate Organic Matter in the Ocean

This paper presents a review of the past decade's highlights of research on the isolation and characterisation of particulate organic matter (POM) in the world's oceans. The emphasis is on chemical studies but, in keeping with the growing interdisciplinary nature of marine science, advances in other disciplines are also discussed, particularly those in biological sciences. Increasing evidence for the importance of picoplankton, bacteria and viruses as POM constituents is highlighted, including the recent recognition of large populations of autotrophic bacteria able to harvest light for energy. The transport of POM to bottom waters was thought to be largely confined to large, rapidly sinking faecal pellets. However, recent studies have highlighted the importance of organic aggregates and flocs formed by diatoms such as Rhizosolenia and other microalgae. Ascending particles have also been discovered, many of which are lipid-rich. Several studies have shown that resuspension of bottom sediments and lateral advection of material from continental shelves can lead to anomalously high particle fluxes measured in sediment traps moored in deep water. Many new approaches for characterizing POM have emerged, such as pyrolysis gas chromatography-mass spectrometry and direct temperature-resolved mass spectrometry for analysis of higher molecular weight materials and biopolymers. Lipid biomarker techniques have also advanced, exciting new possibilities being raised by the ability to measure stable and radioactive carbon isotopes for individual compounds. The techniques of molecular biology, such as the polymerase chain reaction (PCR), are being increasingly applied to provide complementary information to more conventional microscopy and flow cytometry on the identity of organisms in the sea. The combination of these techniques with advanced chemical analysis methods promises to greatly increase our knowledge of the origins, transport and fate of organic matter in the oceans. This revised version was published online in August 2006 with corrections to the Cover Date.     

厚壳贻贝(Mytilus coruscus)中抗生素耐受细菌多样性研究

抗生素的滥用是当前水产养殖业发展及食品安全面临的重大威胁。贻贝体内含有丰富的附生菌群,但目前尚不清楚贻贝附生菌群在抗生素残留背景下的动态变化和耐受性特征。为此进行了厚壳贻贝中抗生素耐受细菌多样性研究。将厚壳贻贝暴露于含有青霉素、链霉素和卡那霉素的养殖水体中,之后采用超声法获取含贻贝软体部组织表面附着菌群的超声液,经Zobell 2216E液体培养基培养后,将组织超声液和培养液一并经16S rDNA高通量测序来分析厚壳贻贝微生物的群落组成。结果表明,超声处理后获得的微生物主要为拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)和疣微菌门(Verrucomicrobia)。多重抗生素的处理可以显著下降贻贝体内微生物的OTU数,对微生物群落结构有明显影响。经264h恢复养殖后,在门水平上观察到贻贝体内微生物群落可以恢复重建,表明其具有一定的韧性。研究结果将为今后进一步探索宿主相关微生物群落构建机制及特殊抗性微生物奠定基础。     

水产品鲐鱼(Pneumatophorus japonicus)细菌多样性及优势微生物分析

为研究鲐鱼(Pneumatophorus japonicus)在不同冻藏时期的优势微生物,并且得到一个较全面的细菌群落组成结构,采用16SrDNA基因克隆文库及克隆子分析等方法,分别对冻藏1d、8d和20d条件下鲐鱼鳃体系中的细菌种群多样性、优势微生物及其中产组胺菌的变化进行分析。结果表明,不同时期鲐鱼鳃内的细菌多样性均较强;冻藏1天的鳃体系中不动杆菌属(Acinetobacter)占优势为总数的77.5%,产组胺细菌如泛菌属(Pantoea)和肠杆菌属(Enterobacter)较少;第8天优势细菌种群是梭杆菌属(Fusobacteria bacterium)和希瓦氏菌属(Shewanella),而产组胺菌如发光杆菌属(Photobacterium)和弧菌属(Vibrio)较前期数量有所增加;冻藏到第20天的优势种群为哈夫尼菌属(Hafnia)和沙雷菌属(Serratia),它们均是产组胺的菌群。鲐鱼中细菌多样性及其优势微生物的变化等全面信息可为了解鲐鱼水产品组胺发生的微生物机制及其控制方法提供重要依据。     

海洋球石藻(Emiliania huxleyi)通用表达载体的构建与电转化

海洋球石藻Emiliania huxleyi是一种全球广泛分布的真核浮游植物,该种不仅是海洋碳、硫循环和全球气候变化的重要指示物种,而且能够产生丰富的次级代谢生物活性物质,在生物技术领域也具有很好的应用前景。本文通过分析氨苄青霉素、卡那霉素、G418、氯霉素、链霉素、新生霉素及嘌呤霉素等7种常用抗生素对海洋球石藻生长的影响,确定G418可作为该藻阳性转化藻株的抗性筛选试剂,其对应的抗性基因neo则作为该藻表达载体构建中的抗性筛选标记。在此基础上克隆了绿色荧光蛋白基因gfp、抗性标记基因neo及E. huxleyi BOF92内源性岩藻黄素-叶绿素a/c结合蛋白基因的启动子fcp,以pUC18为基础载体,构建了pUC18-fcp-gfp和pUC18-fcp-neo两个重组表达载体,以电转化方法共转化球石藻细胞并结合选择性固体培养基筛选,成功获得了被转化的球石藻细胞。海洋球石藻遗传转化系统的建立为进一步开展该种相关的基础生物学研究及其在生物技术领域的应用奠定了基础。     

深海热液喷口金属硫化物中甲烷菌的多样性研究

对两个来自太平洋洋脊胡安·德富卡的两个热液烟囱的金属硫化物4136-2和4148-B1进行甲烷菌甲基辅酶 M 还原酶的编码基因 mcrA 的序列扩增,构建克隆文库并进行分子进化分析.结果表明在这个甲烷富集的热液喷口周围含有丰富的甲烷产生菌,没有任何甲烷氧化菌存在.两个硫化物样品的甲烷产生菌种类完全不同.在4136-2硫化物中的甲烷产生菌都与热液口的高温环境有关系,主要属于甲烷球菌目的甲烷暖球菌(Methanocaldococcus),少部分属于甲烷火菌目甲烷嗜高热菌属的坎氏甲烷嗜高热菌(Methanopyrus kandleri).与这两个属的可分离菌株的氨基酸同源性为89%~97%,核苷酸同源性高达92%~100%.4148-B1硫化物中发现的一类疑似甲基辅酶 M 还原酶的编码序列,它们与已知的甲烷菌 mcrA 序列核苷酸同源性为69%~72%,氨基酸同源性仅为43%~47%.这可能是由于4148-B1来自于正在喷发的超高温热液喷口相关.由于与已发表的甲烷菌克隆子或菌株同源性较低,有可能是热液口特有的以前未发现的甲烷菌     

基于高通量测序技术解析鲣鱼(Katsuwonus pelamis)鱼油和鱼白调控肠道菌群结构作用

本研究利用高通量测序技术分析小鼠喂食鲣鱼(Katsuwonus pelamis)鱼油和鱼白酶解液后肠道微生物的多样性、群落组成以及结构的变化,探讨高蛋白、高脂肪饮食对人类健康的影响。研究结果显示:喂食鱼油组小鼠的肠道微生物丰富且多样。在门水平上,对照组和鱼油组的优势菌门为拟杆菌门(Bacteroidetes);鱼白组的优势菌门为厚壁菌门(Firmicutes)。在科水平上,对照组和鱼油组中的优势菌科为Muribaculaceae;鱼白组中的优势菌科为毛螺菌科(Lachnospiraceae)。在属水平上,对照组和鱼油组中的优势菌属为Muribaculum;鱼白组中的优势菌为Lachnoclostridium;对照组、鱼油组和鱼白组小鼠的肠道微生物群落中瘤胃球菌属(Ruminococcus)和拟杆菌属(Bacteroides)等丰度较高。在种水平上,对照组和鱼油组中的优势菌为Muribaculum intestinale;鱼白组中的优势菌为解糖梭菌(Clostridium saccharolyticum)。研究结果证明鲣鱼鱼油和鱼白可以调节菌群结构和组成,增加肠道有益菌,减少病原菌的丰度等。     

Doubled CO2 impact on subarctic marine carbon cycle: a case study at Ocean Station P

Along with meteorological observations, complementary and systematic oceanographic observations of various physical, biological and chemical parameters have been made at Ocean Station P (OSP) (50°N, 145°W) since the early 1950s. These decadal time scale data have contributed to a better understanding of the physical, biological and chemical processes in the surface layer of the northeastern subarctic region of the Pacific Ocean. These data have demonstrated the importance of the North Pacific in the global carbon cycle and, in particular, the role of biological/chemical processes in the net exchange of CO₂ across the air–sea interface. Although we do not fully comprehend how climatic variations influence marine communities or marine biogeochemistry, previous studies have provided some basic understanding of the mechanisms controlling the seasonal and inter-annual variations of biological and chemical parameters (such as phytoplankton, bacteria, nitrate/ammonium concentration) at OSP, and how they affect the carbon cycling in the subarctic North Pacific. In this study, we investigate how these mechanisms might alter the seasonal variations of these parameters at OSP under a 2XCO₂ condition. We examine these influences using a new biological model calibrated by the climatological data from OSP. For the 2XCO₂ simulation, the biological model is driven off line (i.e., no feedback to the ocean/atmospheric model components) by the climatology plus 2XCO₂−1XCO₂ outputs from a global surface ocean model and the Canadian GCM. Under the 2XCO₂ condition, the upper layer ocean shows an increase in the entrainment rate at the bottom of the mixed layer for OSP during the late autumn and winter seasons, resulting in an increase in the f-ratio. Although there is an overall increase in the primary production (PP) by 3–18%, a decrease in the biomass of small phytoplankton and microzooplankton (due to mesozooplankton grazing) lowers the concentration of dissolved organic matter (DOM) by 4–25%. The model also predicts a significant increase in the concentrations of nitrate and ammonium, and in bacterial production during July and August. Doubling of the atmospheric CO₂ from 330 to 660 ppm forces the marine pCO₂ to increase by about 63%, much of which is driven by an increased flux of CO₂ from the atmosphere to the oceans.     

几丁聚糖生物敷料的研究与开发

以几丁聚糖为主要原料,采用理化、正交等试验,研制了几丁聚糖生物敷料配方和生产工艺,并通过体外抗菌、皮肤刺激、皮肤致敏试验,对其细胞毒性及动物药效学等进行药理检验和安全性评价。研究结果表明,使用几丁聚糖生物敷料安全有效,几丁聚糖生物敷料具有良好的生物相容性和天然、广谱抗菌活性,同时还具有明显保护创面、预防感染、促进愈合等生物学功效。     

抗生素对浒苔形态及共附生细菌多样性的影响

本文研究了3种抗生素联合处理黄海绿潮浒苔(Ulva prolifera)形态及共附生细菌多样性的影响。将浒苔放散的配子加入到含有3种广谱抗生素(青霉素G、硫酸新霉素、多黏菌素B)的培养液中进行培养, 与对照组相比, 发现浒苔幼苗生长速度缓慢, 且细胞变小, 藻体无法形成正常形态。进一步采用16S rDNA高通量测序分析抗生素处理藻体前后的共附生细菌多样性, 发现抗生素处理后浒苔共附生细菌丰富度和多样性降低, 其中Maribacter丰度降低。将藻体与形态发生化合物thallusin(Maribacter spp. 提取物)共培养, 发现thallusin可促进藻体形态恢复和生长, 推测抗生素可通过抑制Maribacter等细菌从而影响浒苔形态和生长, 这将为浒苔共附生细菌功能的深入研究奠定一定基础。     

Thermophilic subseafloor microorganisms from the 1996 North Gorda Ridge eruption

High-temperature microbes were present in two hydrothermal event plumes (EP96A and B) resulting from the February–March 1996 eruptions along the North Gorda Ridge. Anaerobic thermophiles were cultured from 17 of 22 plume samples at levels exceeding 200 organisms per liter; no thermophiles were cultured from any of 12 samples of background seawater. As these microorganisms grow at temperatures of 50–90°C, they could not have grown in the event plume and instead most probably derived from a subseafloor environment tapped by the event plume source fluids. Event plumes are thought to derive from a pre-existing subseafloor fluid reservoir, which implies that these thermophiles are members of a native subseafloor community that was present before the eruptive event. Thermophiles also were cultured from continuous chronic-style hydrothermal plumes in April 1996; these plumes may have formed from cooling lava piles. To better understand the nutritional, chemical, and physical constraints of pre-eruptive crustal environments, seven coccoidal isolates from the two event plumes were partially characterized. Results from nutritional and phylogenetic studies indicate that these thermophiles are heterotrophic archaea that represent new species, and probably a new genus, within the Thermococcales.     

单环刺螠育苗养殖及综合利用研究进展

单环刺螠(Urechis unicinctus)是我国北方沿海常见的一种海洋底栖无脊椎动物,也是具有较好市场开发前景的新型海水养殖品种之一。本文概述了单环刺螠的基础生物学、人工育苗和养殖现状,以及高附值开发潜力,并提出了构建单环刺螠新型海水养殖产业链的可行性。     

海洋中单环芳烃(BTEX)的来源、分布与迁移及其生态环境效应

挥发性有机物排放是目前大气环境污染的一个焦点问题,以苯、甲苯、乙苯和邻/间/对二甲苯(BTEX)为主要代表的单环芳烃是其中一类重要的化合物。受人类生产活动的影响,BTEX在大气、土壤、地表及地下径流、海洋中普遍存在,其环境效应与生态毒害受到广泛关注。本文从大气-海水-沉积物-生物体入手,综述了海洋环境中BTEX的分布、来源及其迁移过程等研究进展。研究显示海洋环境中BTEX浓度变化范围较大,在分布上呈现出近海高、远洋低的趋势。海洋中BTEX的主要来源是人为来源,即海洋是陆源BTEX重要的汇,而海-气交换和生物降解则被认为是BTEX的主要去除途径。此外,本文阐释了海洋中BTEX的生态与大气环境效应。BTEX具有生物毒性,威胁海洋生物的生存与繁衍,破坏海洋生态系统平衡;而进入大气的BTEX能够影响大气的氧化能力,对于臭氧和二次有机气溶胶的形成具有重要贡献。尽管海洋BTEX研究取得了一定的认识,但目前海洋环境不同介质中BTEX来源解析、迁移转化过程及生物毒性机理亟需深入系统研究,为深入认识BTEX的生物地球化学过程与环境效应奠定基础。     

Antibiotic resistance in coliform and faecal coliform bacteria from natural waters and effluents

Coliform and faecal coliform bacteria isolated from potable water supplies (springs and wells), streams, seawater, and effluents were examined for the incidence of antibiotic resistance. More than 40% of the coliform and faecal coliform bacteria isolated from drinking water supplies were resistant to one or more antibiotics. In stream and seawater samples, the incidence was increased to more than 60%, and to more than 70% among faecal coliforms isolated from effluents. Approximately 40% of the isolates resistant to streptomycin or tetra‐cycline were able to transfer their resistance determinants to an antibiotic‐sensitive strain of Escherichia coli within 2 h.

Although the widespread use of antibiotics is perhaps the major selective force favouring the increased incidence of antibiotic‐resistant bacteria, antibiotic‐resistant faecal coliforms may also have a selective advantage in natural waters.     

Overlapping distributions of cryptic Scoloplos cf. armiger species in the western Wadden Sea

Unknowingly lumping cryptic species in biological monitoring studies hinders progress in understanding their functioning in a wide range of research fields including population dynamics, ecophysiology and community ecology. The common polychaete worm Scoloplos cf. armiger is a cryptic species complex comprising entirely different developmental modes: holobenthic and pelago-benthic development. In the northeast Atlantic, three putative species have been described on the basis of molecular data and a breeding study. We report on the development of a fast genotyping assay and on the occurrence and distribution of different molecular types in the western Wadden Sea. The genotyping assay consists of PCR-RFLP analysis by two enzymes of a mitochondrial (cox3-trnQ-nad6) DNA segment. A new, fourth type was observed and this was the only type whose geographical distribution was not uniform but instead skewed toward the eastern part of the study area. All three hitherto known types were also observed and these three displayed a significant difference in depth distribution within the study area. This is the first ecological difference reported for the ‘Subtidal Clade’ (SC) versus the ‘Type Locality Clade’ (TLC). The new type (‘Intertidal Clade 2’) had a similar distribution to what is known as the Intertidal Clade (IC). However, the most striking observation is the large degree of overlap between the depth distributions in the western Wadden Sea, where the clade names ‘Intertidal’ and ‘Subtidal’ do not necessarily reflect local species composition. The necessity to genotype Scoloplos cf. armiger in monitoring programs is emphasized.