Effects of temperature on cellular and biochemical parameters in the crab Carcinus aestuarii (Crustacea, Decapoda)

The effects of temperature on cellular and biochemical parameters of the crab Carcinus aestuarii were evaluated. Crabs were kept for 7 days at 4, 17 (reference value) and 30 °C (salinity of 35 psu), and total haemocyte count (THC), haemocyte volume, haemocyte proliferation, phenoloxidase (PO) activity in both haemocyte lysate (HL) and cell-free haemolymph (CFH), CFH total protein and glucose levels, superoxide dismutase (SOD) and catalase (CAT) activities in both gills and digestive gland were evaluated. The lowest and the highest temperature significantly decreased THC, whereas haemocyte volume and haemolymph glucose concentration did not differ significantly among experimental conditions. Haemolymph protein concentration significantly reduced in crabs maintained at 30 °C, when compared with that of animals kept at 4 and 17 °C. Haemocyte proliferation increased significantly in crabs kept at 4 and 30 °C, when compared with that of crabs held at 17 °C. Likewise, a significantly higher PO activity was recorded in CFH from crabs kept at 4 and 30 °C, than in control crabs. Conversely, PO activity did not vary significantly in HL. With regard to antioxidant enzyme activities, a significant decrease in CAT activity was observed in gills from crabs kept at 4 °C, when compared to that of crabs kept at 17 °C and 30 °C. Results obtained demonstrated that the highest and lowest temperature tested influenced crab biological responses, and indicated that C. aestuarii modulated its cellular and biochemical parameters (mainly haemocyte proliferation, CFH protein concentrations and CFH PO activity) in order to cope with temperature.     

Uptake and release of paralytic shellfish toxins by the clam Ruditapes decussatus exposed to Gymnodinium catenatum and subsequent depuration

A laboratory experiment was performed with the clam Ruditapes decussatus, fed with the toxic dinoflagellate Gymnodinium catenatum and the non-toxic algae Isochrysis galbana (14 days) and subsequently only with I. galbana (15 days). Individual paralytic shellfish toxins were determined by LC-FLD in G. catenatum cells, whole clam tissues, and particulate organic matter (POM) produced by clams. The toxins dcSTX and dcGTX2 + 3 in the algae were less abundant than C1 + 2 and B1, but were predominant in clams during both the exposure and depuration phases. The toxin dcNEO was only detected in clams during a short period, indicating conversion from other compounds. The toxin composition of the POM indicated the export of dcSTX as faeces or pseudo-faeces along the entire experiment (2.5–14 nmol mg⁻¹), B1 was present in a short period of the exposure and C1 + 2 and dcGTX2 + 3 absent. A mass balance calculation indicated that approximately 95% of C1 + 2 and 85% of B1 supplied to the clams were converted into other toxins or lost in solution. Conversely, the net gain of 512, 61 and 31 nmol for dcSTX, dcGTX2 + 3 and dcNEO, respectively, suggests the conversion from other assimilated compounds by clams during exposure and depuration phases.     

A multi-biomarker approach to assess effects of Triclosan in the clam Ruditapes philippinarum

The effects of 7 days’ exposure to differing Triclosan (TCS) concentrations (300, 600, and 900 ng/L) were investigated in the clam Ruditapes philippinarum. Vitellogenin (Vg)-like protein levels in haemolymph and digestive gland from males and females, gill acetylcholinesterase (AChE) activity, superoxide dismutase (SOD) and catalase (CAT) activities in gills and digestive gland, and gill lipid peroxidation (LPO) were measured. The highest TCS concentrations decreased significantly Vg levels in male haemolymph and digestive gland, whereas no significant variations were found in females. The highest TCS concentrations increased significantly SOD activity in gills, but decreased it in digestive gland. No changes in CAT activity were observed. In gills, TCS reduced significantly AChE activity, but it did not induce significant variations in LPO. Our study demonstrates that TCS alters biochemical parameters in R. philippinarum, even at environmentally realistic concentrations, and suggests differing modes of action of the contaminant, in clams at least.     

Seasonal and gender-related differences in morphometric features and cellular and biochemical parameters of Carcinus aestuarii from the Lagoon of Venice

In this study, the seasonal variations in the morphometric features and in the cellular and biochemical parameters of the haemolymph were investigated in both male and female crabs (Carcinus aestuarii). Crabs were seasonally (November 2010–August 2011) collected from the Lagoon of Venice, and the moult stage, weight, width and length of the carapace, and width and length of the bigger chela were evaluated. In addition, the total haemocyte count (THC), haemocyte diameter and volume, haemolymph glucose and total protein levels, and haemolymph phenoloxidase (PO) and N-acetyl-β-glucosaminidase (NAG) activities were measured. The results demonstrated that the collected crabs were all in the intermoult stage and that the males were bigger than the females. A two-way ANOVA revealed a significant effect of season on the THC and the haemocyte volume and a significant influence of gender on the haemocyte diameter. Season and gender significantly affected the haemolymph glucose concentration, whereas haemolymph protein levels were dependent only on the season. In addition, both season and gender significantly influenced the PO and NAG activities in the haemolymph. Overall, the results demonstrated that crab morphometric features as well as haemolymph cellular and biochemical parameters varied markedly as a function of both season and gender.     

Speciation of Fe in the Eastern North Atlantic Ocean

In the Eastern North Atlantic Ocean iron (Fe) speciation was investigated in three size fractions: the dissolvable from unfiltered samples, the dissolved fraction (<0.2 μm) and the fraction smaller than 1000 kDa (<1000 kDa). Fe concentrations were measured by flow injection analysis and the organic Fe complexation by voltammetry. In the research area the water column consisted of North Atlantic Central Water (NACW), below which Mediterranean Overflow Water (MOW) was found with the core between 800 and 1000 m depth. Below 2000 m depth the North Atlantic Deep Water (NADW) proper was recognised. Dissolved Fe and Fe in the <1000 kDa fraction showed a nutrient like profile, depleted at the surface, increasing until 500–1000 m depth below which the concentration remained constant. Fe in unfiltered samples clearly showed the MOW with high concentrations (4 nM) compared to the overlying NACW and the underlying NADW, with 0.9 nM and 2 nM Fe, respectively. By using excess ligand (Excess L) concentrations as parameter we show a potential to bind Fe. The surface mixed layer had the highest excess ligand concentrations in all size fractions due to phytoplankton uptake and possible ligand production. The ratio of Excess L over Fe proved to be a complementary tool in revealing the relative saturation state of the ligands with Fe. In the whole water column, the organic ligands in the larger colloidal fraction (between 0.2 μm and 1000 kDa) were saturated with Fe, whereas those in the smallest fraction (<1000 kDa) were not saturated with Fe, confirming that this fraction was the most reactive one and regulates dissolution and colloid aggregation and scavenging processes. This regulation was remarkably stable with depth since the alpha factor (product of Excess L and K′), expressing the reactivity of the ligands, did not vary and was 10¹³. Whereas, in the NACW and the MOW, the ligands in the particulate (>0.2 μm) fraction were unsaturated with Fe with respect to the dissolved fraction, thus these waters had a scavenging potential.     

Implications of seasonal priming and reproductive activity on the interpretation of Comet assay data derived from the clam, Tapes semidecussatus Reeves 1864, exposed to contaminated sediments

We explore the use of the clam Tapes semidecussatus Reeves 1864 as an indicator for the presence of potentially genotoxic substances in estuarine sediments. The limitations associated with the interpretation of Comet assay data (expressed as % DNA in tail) in terms of clam reproductive state, size (age) and thermal exposure history following laboratory acclimation are discussed. Hatchery-reared clams, subjected to ambient temperature fluctuations during growth, were exposed in vivo under laboratory conditions for three weeks to sediment samples collected from a polluted site and a "clean" reference site. The DNA damage observed in haemocytes, gill and digestive gland cells was significantly higher in animals exposed to contaminated sediment compared to those exposed to sediment from the reference site. The extent of DNA damage recorded was not correlated with size (age). Spawning was not observed during the experiment. Nevertheless, clams with well-developed gonads showed a statistically higher degree of DNA damage in gill and digestive gland cells- but not haemocytes, demonstrating an increased sensitivity to potential genotoxic compounds, possibly caused by impaired DNA repair capacity due to reproductive activity. Furthermore, the degree of DNA damage in clams exposed to contaminated sediments was higher in autumn and winter compared to spring and summer, suggesting an effect of seasonal priming.     

Application of primary haemocyte culture of Penaeus monodon in the assessment of cytotoxicity and genotoxicity of heavy metals and pesticides

Lack of shrimp cell lines has hindered the study of pollutants which adversely affects shrimp health and its export value. In this context a primary haemocyte culture developed from Penaeus monodon was employed for assessing the cytotoxicity and genotoxicity of two heavy metal compounds, cadmium chloride and mercuric chloride and two organophosphate insecticides, malathion and monocrotophos. Using MTT assay 12?h IC(50) values calculated were 31.09?±?16.27?μM and 5.52?±?1.16?μM for cadmium chloride and mercuric chloride and 59.94?±?52.30?mg?l(-1) and 186.76?±?77.00?mg?l(-1) for malathion and monocrotophos respectively. Employing Comet assay, DNA damage inflicted by these pollutants on haemocytes were evaluated and the pollutants induced DNA damage in >60% of the cells. The study suggested that haemocyte culture could be used as a tool for quantifying cytotoxicity and genotoxicity of aquaculture drugs, management chemicals and pollutants.     

Can starvation influence cellular and biochemical parameters in the crab Carcinus aestuarii?

Crustacea experience periods of starvation during moulting or when limited food availability occurs. The effects of starvation on Crustacea physiological responses have been demonstrated, whereas the effects of starvation on Crustacea immune parameters remain to be more fully studied. In the present study the effects of starvation on immune parameters and antioxidant enzyme activities of the crab Carcinus aestuarii were evaluated for the first time. Treated crabs were starved for 7 days, whereas control crabs were fed daily with mussels. Total haemocyte count (THC), haemocyte diameter and volume, haemocyte proliferation, cell-free haemolymph (CFH) glucose and total protein levels, and phenoloxidase (PO) activity in both haemocyte lysate (HL) and CFH were measured in crabs. In addition, superoxide dismutase (SOD) and catalase (CAT) activities were evaluated in both gills and digestive gland from crabs, in order to evaluate whether starvation induced oxidative stress in C. aestuarii. THC increased significantly in starved crabs, with respect to controls, whereas no significant variations were observed in haemocyte diameter, volume and proliferation. In CFH of starved animals glucose concentration significantly increased, whereas total protein concentration significantly reduced. A significantly higher PO activity was recorded in HL from starved crabs, than in control crabs. Conversely, PO activity did not vary significantly in CFH. Starvation did not cause significant alterations in antioxidant enzyme activities in both gills and digestive gland. Results obtained demonstrated that starvation influenced crab immune parameters, but did not induce oxidative stress. Results also indicated that C. aestuarii can modulate its cellular and biochemical parameters in order to cope with starvation.     

Size-fractionated biological iron and carbon uptake along a coastal to offshore transect in the NE Pacific

Iron has been shown to limit phytoplankton growth in high-nutrient low-chlorophyll (HNLC) regions such as the NE subarctic Pacific. We report size-fractionated Fe-uptake rates by the entire plankton community in short (6–8 h) light and dark incubations along an E–W transect from P04 (a coastal ocean station) to OSP (an open-ocean HNLC station) during August–September 1997. Size-fractionated primary productivity and chl a were measured to monitor algal Fe : C uptake ratios and Fe-uptake relative to phytoplankton biomass. The >5.0 μm size-class, which consisted mostly of large diatoms, had the highest Fe-uptake rate at nearshore stations (P04 and P8), but Fe-uptake rates for this size class decreased despite increases in biomass and primary productivity when transecting westwards to HNLC waters. Fe-uptake rates of the small size class (0.2–1.0 μm, including heterotrophic bacteria and autotrophs) were inversely related to the >5.0 μm size-class uptake rates, in that stations with high dissolved Fe (DFe) concentrations had relatively low uptake rates compared to those in the low-Fe offshore region. The 1.0–5.0 μm size-class Fe-uptake rates were low, relatively invariant along the transect, and differed little between light and dark incubations. Dark Fe-uptake rates averaged 10–20% less than those in the light for the >5.0 μm size class. Dark uptake rates however, were higher than light uptake rates for the 0.2–1.0 μm size class at all stations. Fe : C uptake ratios were high for all size classes at P04, but decreased as DFe concentrations decreased offshore. The prokaryote-dominated 0.2–1.0 μm size class had the highest Fe : C uptake ratios at all stations. These data suggest that prokaryotic organisms make an important contribution to biological Fe uptake in this region. Our experiments support the results of previous culture work, suggesting higher Fe : C ratios in coastal phytoplankton compared to open-ocean species, and demonstrate that light can have a large effect on Fe partitioning between size classes in subarctic Pacific HNLC waters.     

凡纳滨对虾(Litopenaeus vannamei)感染白斑症病毒(WSSV)后大颗粒血细胞的变化

采用抗凡纳滨对虾大颗粒细胞的单克隆抗体，应用免疫细胞化学方法，分析了凡纳滨对虾人工感染白斑症病毒(WSSV)后血细胞中大颗粒细胞的变化。研究表明，凡纳滨对虾感染WSSV后，大颗粒细胞的细胞质较正常细胞的多；从WSSV感染凡纳滨对虾的第1天至第3天，大颗粒细胞的数量增加幅度较大。同时，大颗粒细胞占血细胞总数的比例也增加较大：第1天为15．2％，第2天为18％，第3天增大到45．9％，而后大颗粒细胞的比例稍有下降，到第8天下降到43．3％，但仍明显高于对照组(健康虾的大颗粒细胞比例为15．2％)。     

DNA strand breakage in mussels (Mytilus edulis L.) deployed in intertidal and subtidal zone in Reykjavík harbour

DNA single-strand breaks were analysed in the blue mussel (Mytilus edulis L.) deployed in intertidal and subtidal zones in the PAH contaminated Reykjavík harbour and at a reference site, Hvalfj?rethur, Iceland. DNA strand breaks were analysed by Comet assay in isolated gill and haemocyte cells from six mussels from each site and depth. Increased DNA damage in both gill cells and haemocytes were observed in mussels deployed in Reykjavík harbour compared to the reference site. Intertidal mussels from Reykjavík harbour had higher DNA damage in haemocytes compared to subtidal mussels. The Comet assay seems to be useful for measuring genotoxic exposure in mussels from the field, and that DNA damage might be higher in the intertidal zone either due to higher exposure to contaminants or because of physiological and biochemical responses to variations in oxygen availability.     

低溶解氧对凡纳滨对虾酚氧化酶原激活系统和相关免疫因子的影响

本文研究了凡纳滨对虾在低溶解氧条件下酚氧化酶原系统的激活机制。结果表明:低溶氧(1.5mg/L,3.0mg/L)胁迫对凡纳滨对虾血淋巴中多巴胺含量、血细胞数量、酚氧化酶原激活系统相关酶活力、血淋巴中类α2-巨球蛋白活力影响显著(P<0.05),且各指标表现出明显的时间效应性。在实验时间内,DO为3.0mg/L处理组,总血细胞、小颗粒细胞和透明细胞数量均在12h内降低而后趋于稳定,大颗粒细胞数量在24h内降低后保持稳定,DO为1.5mg/L处理组,总血细胞数量和三类血细胞数量均在24h内降低而后趋于稳定;DO为3.0mg/L和1.5mg/L处理组凡纳滨对虾血淋巴中多巴胺含量分别在24h、12h内呈峰值变化,至12h、6h时达到最大值,在24h、12h后恢复至对照组水平并趋于稳定;血细胞中酚氧化酶原活力分别在12h、6h时显著降低,且均在24h时趋于稳定;丝氨酸蛋白酶活力在6h时均显著降低,在24h时达到最低值,而后趋于稳定,蛋白酶抑制剂活力显著降低,但类α2-巨球蛋白活力48h内呈峰值变化,与对照组比较均显著升高,48h后趋于稳定。实验期内对照组DO为5.5mg/L各指标变化不显著。     

Anchoring metabolic changes to phenotypic effects in the chlorophyte Scenedesmus vacuolatus under chemical exposure

In order to derive a causal understanding of toxic effects in organisms, ecotoxicology may benefit from linking molecular changes, evaluated by ‘omics’-techniques, to phenotypic observations. However, an approach to link these observation levels is still lacking.The aim of this study was to relate metabolic changes in the chlorophyte Scenedesmus vacuolatus to established parameters of toxicity. Therefore, synchronized cultures of the alga were exposed for 14 h to the phytotoxicant N-phenyl-2-naphthylamine (PNA) in the range of 0.00089 μmol L⁻¹ (environmental concentrations) up to 1.82 μmol L⁻¹. Cell growth and photosynthesis inhibition were evaluated but revealed no effect of PNA at experimental concentrations below 0.456 μmol L⁻¹. Changes in the biochemical composition of algae were measured by GC–MS in both polar and non-polar phases. PCA uncovered no separation in the multivariate pattern of mass spectral features at exposure concentrations below 0.00356 μmol L⁻¹ of PNA. However, a clear separation was detected at concentrations higher than 0.00713 μmol L⁻¹. A combined visualization of PCA results for metabolic changes and concentration–response relationships for growth and photosynthesis inhibition revealed (I) a two orders of magnitude higher sensitivity of metabolomics to detects changes after PNA exposure compared to the phenotypic parameters measured and (II) two types of metabolic responses: one group of features was reflecting pharmacological effects at low exposure concentrations and the second group corresponded to adverse effects along with conventional observations of toxicity.     

Effect studies of human pharmaceuticals on Fucus vesiculosus and Gammarus spp

In two experiments, the human pharmaceutical propranolol negatively affected the physiology of two test organisms, Fucus vesiculosus and Gammarus spp. from a Baltic Sea littoral community in a concentration of 1000 μg l⁻¹. Some effects were also observed for the lower, more ecologically relevant concentrations (10 μg l⁻¹ and 100 μg l⁻¹). The effects on F. vesiculosus not only increased with increasing concentration, but also with exposure time; while the effects on Gammarus spp. were more inconsistent over time. No clear effects of the pharmaceuticals diclofenac and ibuprofen were observed for any of the organisms. Physiological parameters measured were GP:R-ratio, chlorophyll fluorescence and release of coloured dissolved organic matter, respiration and ammonium excretion. Pharmaceutical substances are repeatedly detected in the Baltic Sea which is the recipient for STP effluents from more than 85 million people living in the catchment area, but the knowledge of their effects on non-target organisms is still very limited.     

Bioavailability and effects of sediment-bound TBT in deposit-feeding clams, Scrobicularia plana

Concentrations of tributyltin (TBT), dibutyltin (DBT), inorganic and total tin have been measured in water, sediments and deposit feeding clams, Scrobicularia plana, from 25 estuarine locations in England and Wales, in order to compare the bioavailability of different forms of the element. Abnormally high tin values in Scrobicularia from harbours and areas of high boating activity testify to the high bioaccumulation potential of organotins, notably TBT derived from anti-fouling paints, relative to inorganic tin.Infaunal deposit-feeding bivalves consistently contain higher concentrations of TBT than other benthic organisms indicating that sediments may be an important route for uptake. Significant correlations between sediment TBT values and residues in clams such as S. plana (r = 0·81, p < 0·001) support this hypothesis. Direct evidence of the bioaccumulation of sediment-bound TBT is provided from laboratory experiments in which Scrobicularia were exposed to TBT in water and in sediments, either separately or in combination. The results of these experiments clearly demonstrate a predominantly particulate component for TBT uptake in clams.The kinetics of TBT accumulation and elimination were studied in Scrobicularia exposed to sediments containing 0·3 and 1·0 μg/g TBT (as tin). Equilibrium concentrations in tissue are approached after 40 days.Sediments containing 10 μg/g TBT are acutely toxic to S. plana, although such concentrations are only likely to occur close to dockyards and large marinas. Preliminary laboratory and field observations suggest however, that clam populations could be affected at TBT concentrations in sediments of 0·3 μg/g and possibly lower.     

Effect of macronutrient enrichment on the size distribution, sorption, and bioconcentration factor of iron by coastal phytoplanktonic diatoms

The influences of macronutrient enrichment on iron absorption, adsorption, and the bioconcentration factor (BCF) of iron by coastal diatoms were examined. In addition, the distribution of different iron size fractions in seawater in relation to two diatom species exposed to nutrient enrichment over an extended period (6 days for Thalassiosira weissflogii, and 4 days for Skeletonema costatum), was investigated. Iron concentrations were measured of seven size fractions: particles (>0.4 μm), colloidal particles (0.4 μm–100, 100–50, 50–30, 30–10, 10–3 kDa), and soluble species (<3 kDa).The absorption, adsorption, uptake, BCF of iron by the diatoms, and the iron species distribution in seawater were affected by the diatom species, as well as the nitrate (N) and phosphate (P) concentration. The addition of P could affect the iron internalization strategy. The content of soluble iron species was positively correlated with the amount of iron absorption by T. weissflogii, but it was not suitable for S. costatum.     

流式细胞术比较研究4种双壳贝类血细胞的分群

应用流式细胞术对太平洋牡蛎(Crassostrea gigas)、海湾扇贝(Argopecten irradians)、虾夷扇贝(Patinopecten yessoensis)和毛蚶(Scapharca subcrenata)4种双壳贝类的血细胞分类进行了比较。根据其血细胞的前向角散射光(FSC)和侧向角散射光(SSC)特性的不同将血细胞分群,FSC和SSC二维图分析发现太平洋牡蛎、海湾扇贝和虾夷扇贝的血细胞都可分为3个亚群,即透明细胞、小颗粒细胞和大颗粒细胞,而毛蚶的血细胞只可区分为透明细胞和颗粒细胞2个亚群。同时还对这4种双壳贝类各种血细胞所占比例进行了比较研究,发现各亚群细胞所占比例差异很大。     

Effects of okadaic acid on haemocytes from Mytilus galloprovincialis: A comparison between field and laboratory studies

Individuals of Mytilus galloprovincialis, contaminated with Diarrheic Shellfish Poisoning (DSP) toxins, were studied with the aim to correlate the okadaic acid (OA) body burden and the percentage of damaged haemocytes by quantifying annexin V positive cells by flow cytometry. Results showed less percentage of damaged haemocytes in high OA contaminated samples. These data were compared with results of in vitro assays of mussel haemocytes exposed to increased concentrations of OA. Similarly, haemocytes exposed to the most concentrated OA solution were less damaged.     

Influence of chronic exposure to silver and mercury in the field on the bioaccumulation potential of the bivalve Macoma balthica

In order to assess the adaptation to metals previously observed in the bioindicator organism, Macoma balthica, subjected to chronic contamination by silver and mercury in the French Loire estuary, the bioaccumulation potential of individual organisms originating from the contaminated Loire estuary and a relatively uncontaminated control estuary (Somme) was evaluated using both radiotracers and stable isotopes of Ag (80 μg Ag litre⁻¹) and Hg (100 μg Hg litre⁻¹). Clams from the contaminated estuary were more sensitive to Ag (LT₅₀ = 9d) than those originating from the Somme estuary (LT₅₀ > 15d), even though the former bioaccumulated Ag to a significantly lower degree. This is attributed to a consequence of the chronic stress induced by Ag while clams were living in their natural environment. Therefore, past history of trace metal contamination should be considered when evaluating the susceptibility of M. balthica to heavy metal exposure. Lower uptake rates obtained for Hg (during the initial uptake phase only) and for Ag in clams from the polluted estuary suggest the presence of an adaptive trait for survival in contaminated areas. However, the lower degree of bioconcentration observed for Ag was not sufficiently low to reduce the sensitivity of the organisms to Ag and allow them to resist the toxic stress. Clams that survived Ag or Hg exposure at LT₅₀ did not protect themselves against metal toxicity by accumulating a significantly lesser amount of these metals than clams which did not survive metal stress. The results suggest that the bioaccumulation potential of each individual was not a factor which can explain the survival ability of M. balthica exposed to chronic Ag and Hg contamination in estuaries. In this case, cellular, biochemical and genetic levels of adaptation are presumed to be of greater importance.     

Habitat,growth and physiological ecology of a basaltic community of Ridgeia piscesae from the Juan de Fuca Ridge

The vestimentiferan tubeworm Ridgeia piscesae is an ecosystem-structuring organism in the hydrothermal vent environments of the Northeast Pacific. During this study, a single representative aggregation of the long-skinny morphotype of R. piscesae from the main endeavor segment was monitored for 3 yr before being collected in its entirety with a hydraulically actuated collection device manipulated in situ by a research vehicle. Vestimentiferan growth rates in this aggregation were determined by staining the exterior of the tubes and measuring newly deposited tube sections. The average growth rate of R. piscesae in this aggregation was very low in both years of the growth study (3.2 mm yr⁻¹). Although the incidence of plume damage from partial predation was very high (>95%), mortality was very low (<4% yr⁻¹). The distribution and the very tight clustering of recently recruited individuals indicated gregarious settlement behavior that is hypothesized to be partly due to biotic cues from settled larvae. Coupled measurements of vent fluid sulfide concentration and temperature were used to calculate the exposure of the vestimentiferans to sulfide from short- and long-term temperature monitoring. Plume-level temperature records indicate that most of the time individuals in this aggregation were exposed to extremely low levels of vent fluid, and therefore sulfide (<0.1 μM), while their posterior sections were consistently exposed to sulfide concentrations in the 100 μM range. A rootball-like structure formed the common base of the aggregation. In contrast to the anterior sections of the tubeworm tubes, the portions of the tubes within the “rootball” were freely permeable to sulfide. The results of this study show that R. piscesae, unlike vestimentiferans from the East Pacific Rise, can survive and grow in areas of low diffuse vent flow with very low plume-level exposure to sulfide. We propose that this morphotype of R. piscesae has the ability to acquire sulfide from sources near their posterior ends, similar to some species of cold seep vestimentiferans from the Gulf of Mexico. The ability of this single species of vestimentiferan to survive low exposure to vent flow with low mortality coupled with sulfide uptake across posterior tube sections may help explain the occurrence of a single vent vestimentiferan species in a wide variety of habitat conditions at hydrothermal vent sites in the Northeast Pacific.