The Interactive Effects of Elevated CO<Subscript>2</Subscript> and Ammonium Enrichment on the Physiological Performances of <Emphasis Type="Italic">Saccharina japonica</Emphasis> (Laminariales,Phaeophyta)

Environmental challenges such as ocean acidification and eutrophication influence the physiology of kelp species. We investigated their interactive effects on Saccharina japonica (Laminariales, Phaeophyta) under two pH conditions [Low, 7.50; High (control), 8.10] and three NH ₄ ⁺ concentrations (Low, 4; Medium, 60; High, 120 μM). The degree of variation of pH values in the culture medium and inhibition rate of photosynthetic oxygen evolution by acetazolamide were affected by pH treatments. Relative growth rates, carbon, nitrogen, and the C:N ratio in tissue samples were influenced by higher concentrations of NH ₄ ⁺ . Rates of photosynthetic oxygen evolution were enhanced under elevated CO₂ or NH ₄ ⁺ conditions, independently, but these two factors did not show an interactive effect. However, rates of NH ₄ ⁺ uptake were influenced by the interactive effect of increased CO₂ under elevated NH ₄ ⁺ treatment. Although ocean acidification and eutrophication states had an impact on physiological performance, chlorophyll fluorescence was not affected by those conditions. Our results indicated that the physiological reactions by this alga were influenced to some extent by a rise in the levels of CO₂ and NH ₄ ⁺ . Therefore, we expect that the biomass accumulation of S. japonica may well increase under future scenarios of ocean acidification and eutrophication.     

The Effects of Ocean Acidification on Feeding and Contest Behaviour by the Beadlet Anemone <Emphasis Type="Italic">Actinia equina</Emphasis>

Increasing concentrations of atmospheric carbon dioxide are causing oceanic pH to decline worldwide, a phenomenon termed ocean acidification. Mounting experimental evidence indicates that near-future levels of CO₂ will affect calcareous invertebrates such as corals, molluscs and gastropods, by reducing their scope for calcification. Despite extensive research into ocean acidification in recent years, the effects on non-calcifying anthozoans, such as sea anemones, remain little explored. In Western Europe, intertidal anemones such as Actinia equina are abundant, lower trophic-level organisms that function as important ecosystem engineers. Changes to behaviours of these simple predators could have implications for intertidal assemblages. This investigation identified the effects of reduced seawater pH on feeding and contest behaviour by A. equina. Video footage was recorded for A. equina feeding at current-day seawater (pH 8.1), and the least (pH 7.9) and most (pH 7.6) severe end-of-century predictions. Footage was also taken of contests over ownership of space between anemones exposed to reduced pH and those that were not. No statistically significant differences were identified in feeding duration or various aspects of contest behaviour including initiating, winning, inflating acrorhagi, inflicting acrorhagial peels and contest duration. Multivariate analyses showed no effect of pH on a combination of these variables. This provides contrast with other studies where anemones with symbiotic algae thrive in areas of natural increased acidity. Thus, novel experiments using intraspecific contests and resource-holding potential may prove an effective approach to understand sub-lethal consequences of ocean acidification for A. equina, other sea anemones and more broadly for marine ecosystems.     

Retrieval of total ozone in the mesosphere with a new model of electronic-vibrational kinetics of O<Subscript>3</Subscript> and O<Subscript>2</Subscript> photolysis products

The paper presents a new model of electronic-vibrational kinetics of the products of ozone and molecular oxygen photodissociation in the terrestrial middle atmosphere. The model includes 45 excited states of the oxygen molecules O₂(b ¹, Σ _g ⁺ ,v= 0−2), O₂ (a ¹Δ _g , v= 0−5), and O₂(X ³Σ _g ⁻ , v= 1−35) and of the metastable atom O (¹ D) and over 100 aeronomic reactions. The model takes into account the dependence of quantum yields of the production of O₂(a ¹Δ _g , v= 0−5) in a singlet channel of ozone photolysis in the Hartley band on the wavelength of photolytic emission. Taking account of the electronic-vibrational kinetics is important in retrieval of the vertical profiles of ozone concentration from measured intensities of the Atm and IR Atm emissions of the oxygen bands above 65 km and leads to an increase in the ozone concentration retrieved from the 1.27-μm emission, in contrast to the previous model of pure electronic kinetics. Sensitivity analysis of the new model is made for variations in the concentrations of atmospheric constituents ([O₂], [N₂], [O(³P)], [O₃], [CO₂]), the gas temperature, rate constants of the reactions, and quantum yields of the reaction products. A group of reactions that most strongly affect the uncertainty of ozone retrieval from measured intensities of atmospheric emissions of molecular oxygen O₂(b ¹Σ _g ⁺ , v) and O₂(a ¹Δ _g , v) has been determined. Original Russian Text ? V.A. Yankovsky, V.A. Kuleshov, R.O. Manuilova, A.O. Semenov, 2007, published in Izvestiya AN. Fizika Atmosfery i Okeana, 2007, Vol. 43, No. 4, pp. 557–569.     

Effects of temperature on growth,photophysiology, Rubisco gene expression in <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis> and <Emphasis Type="Italic">Karenia mikimotoi</Emphasis>

To explore the effects of temperature changes on dinoflagellate bloom succession in the coastal waters of the East China Sea, changes in the growth, photophysiology, and Rubisco gene expression of Prorocentrum donghaiense and Karenia mikimotoi, two harmful algal species, were investigated at different temperatures (16 to 28°C). The maximal specific growth rate and the maximal mRNA expression of Rubisco gene in P. donghaiense and K. mikimotoi occurred at 20 and 24°C, respectively. The photosynthetic activity of P. donghaiense was generally stable, but K. mikimotoi photosynthesis increased when temperatures rose from 16 to 28°C. The effective photochemical efficiency (F _q ^′ /F _m ^′ ) and the maximal relative electron transfer rate (rETR_max) of K. mikimotoi increased significantly with increasing temperature, and the lowest and highest values occurred at 16 and 28°C, respectively. It seems that P. donghaiense has higher photosynthetic capacity than K. mikimotoi due to its higher F _q ^′ /F _m ^′ , rETR_max, and photosynthetic efficiency (α). However, K. mikimotoi has a higher growth rate than P. donghaiense. These results suggest that the photosynthetic activity and genetic responses of dinoflagellates are species-dependent. It is likely that temperature changes affect species composition during blooms, leading to the observed patterns of bloom succession.     

High mortality and poor growth of green mussels, <Emphasis Type="Italic">Perna viridis</Emphasis>, in high chlorophyll-<Emphasis Type="Italic">a</Emphasis> environment

The current study was carried out from May 2014 to April 2015 to estimate the stock status of P. viridis in Marudu Bay. The gonad development was monitored by histological examination, while the population parameters including asymptotic length (L_∞), growth coefficient (K), mortality rate (Z, F and M), exploitation level (E) and recruitment of P. viridis were estimated using the lengthfrequency data. Results of the current study demonstrated that P. viridis in Marudu Bay spawned throughout the year with two major peaks, one in April to May and another one in October to December. The recruitment pattern was continuous with the peak in May to June 2014, which corresponded to the first spawning peak in April. However, no significant recruitment was observed from the second spawning peak due to the difference in spawning timing between male and female populations. The estimated asymptotic length (L_∞), growth coefficient (K), total mortality (Z), natural mortality (M), fishing mortality (F) and growth performance (ф) of P. viridis in Marudu Bay were estimate to be 117 mm, 0.97 yr^-1, 4.39 yr^-1, 1.23 yr^-1, 3.16 yr^-1 and 4.123, respectively. The exponent b of the lengthweight relationship was 2.4 and exploitation level (E) was 0.72. The high mortality, low condition indices and negative allometric of P. viridis in Marudu Bay is caused by a lack of suitable food in the surrounding water.     

Uptake mechanism of anthropogenic CO<Subscript>2</Subscript> in the Kuroshio Extension region in an ocean general circulation model

The uptake mechanism of anthropogenic CO₂ in the Kuroshio Extension is examined by a Lagrangian approach using a biogeochemical model embedded in an ocean general circulation model. It is found that the uptake of anthropogenic CO₂ is caused mainly by the increase of pCO₂ dependency of seawater on temperature, which is caused by greater dissolved inorganic carbon concentration in the modern state than in the pre-industrial state. In contrast with the view of previous studies, the effect of the vertical entrainment, which brings waters that last contacted the atmosphere with the past lower CO₂ concentration, is comparatively small. Winter uptake of anthropogenic CO₂ increases with the rise of the atmospheric CO₂ level, while summer uptake is relatively stable, resulting in a larger seasonal cycle of the uptake. This increase is significant, especially in the Kuroshio Extension region. It is newly suggested that this increase in the Kuroshio Extension region is largely caused by the combined effects of the increased pCO₂ dependency of the sea water on the temperature and the seasonal difference in cooling.     

Physiological and biochemical responses of <Emphasis Type="Italic">Prorocentrum minimum</Emphasis> to high light stress

Prorocentrum minimum is a common bloomforming photosynthetic dinoflagellate found along the southern coast of Korea. To investigate the adaptive responses of P. minimum to high light stress, we measured growth rate, and generation of reactive oxidative species (ROS), superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) in cultures exposed to normal (NL) and high light levels (HL). The results showed that HL (800 μmol m^?2 s^?1) inhibited growth of P. minimum, with maximal inhibition after 7–9 days. HL also increased the amount of ROS and MDA, suggesting that HL stress leads to oxidative damage and lipid peroxidation in this species. Under HL, we first detected superoxide on day 4 and H₂O₂ on day 5. We also detected SOD activity on day 5 and CAT activity on day 6. The level of lipid peroxidation, an indicator of cell death, was high on day 8. Addition of diphenyleneiodonium (DPI), an NAD(P)H inhibitor, decreased the levels of superoxide generation and lipid peroxidation. Our results indicate that the production of ROS which results from HL stress in P. minimum also induces antioxidative enzymes that counteract oxidative damage and allow P. minimum to survive.     

Age determination and growth rate of <Emphasis Type="Italic">Mactra chinensis</Emphasis> (Bivalvia: Mactridae) by external rings and chondrophore growth bands

Age, growth and mortality of Mactra chinensis were investigated during the period from October 2012 to September 2013 in Busan, South Korea. The monthly variation of the marginal index (MI) of the shell and chondrophore showed that the ring of this species was formed once a year during July. We estimated the age of M. chinensis by reading the external rings on the shell and the growth bands of the chondrophore to compare growth parameters between the two growth characters. The age of this species ranged from 0 to 8 years (shell-based age reading) and from 0 to 10 years (chondrophore-based age reading). Based on external rings and growth bands of chondrophore for the same period, the von Bertalanffy growth functions were expressed by the equation, L _t = 101.53[1-exp {-0.15(t + 0.75)}] and L _t = 90.03[1-exp {-0.20(t + 0.50)}], respectively. The likelihood test showed that there was a significant difference in L _∞ (P < 0.001), K (P < 0.001), to (P < 0.001) estimated from non-linear regression between the two growth characters.     

Ventilation time and anthropogenic CO<Subscript>2</Subscript> in the Bering Sea and the Arctic Ocean based on carbon tetrachloride measurements

The Arctic Ocean is connected to the Pacific by the Bering Sea and the Bering Strait. During the 4th Chinese National Arctic Research Expedition, measurements of carbon tetrachloride (CCl₄) were used to estimate ventilation time-scales and anthropogenic CO₂ (C_ant) concentrations in the Arctic Ocean and Bering Sea based on the transit time distribution method. The profile distribution showed that there was a high-CCl₄ tongue entering through the Canada Basin in the intermediate layer (27.6?<?σ_θ?<?28), at latitudes between 78 and 85°N, which may be related to the inflow of Atlantic water. Between stations B09 and B10, upwelling appeared to occur near the continental slope in the Bering Sea. The ventilation time scales (mean ages) for deep and bottom water in the Arctic Ocean (~?230–380 years) were shorter than in the Bering Sea (~?430–970 years). Higher mean ages show that ventilation processes are weaker in the intermediate water of the Bering Sea than in the Arctic Ocean. The mean C_ant column inventory in the upper 4000 m was higher (60–82 mol m^?2) in the Arctic Ocean compared to the Bering Sea (35–48 mol m^?2).     

Boldness in a deep sea hermit crab to simulated tactile predator attacks is unaffected by ocean acidification

Despite rapidly growing interest in the effects of ocean acidification on marine animals, the ability of deep-sea animals to acclimate or adapt to reduced pH conditions has received little attention. Deep-sea species are generally thought to be less tolerant of environmental variation than shallow-living species because they inhabit relatively stable conditions for nearly all environmental parameters. To explore whether deep-sea hermit crabs (Pagurus tanneri) can acclimate to ocean acidification over several weeks, we compared behavioral “boldness,” measured as time taken to re-emerge from shells after a simulated predatory attack by a toy octopus, under ambient (pH ～7.6) and expected future (pH ～7.1) conditions. The boldness measure for crab behavioral responses did not differ between different pH treatments, suggesting that future deep-sea acidification would not influence anti-predatory behavior. However, we did not examine the effects of olfactory cues released by predators that may affect hermit crab behavior and could be influenced by changes in the ocean carbonate system driven by increasing CO₂ levels.     

Determining the influence of wind-wave breaking on the dissipation of the turbulent kinetic energy in the upper ocean and on the dependence of the turbulent kinetic energy on the stage of wind-wave development

New experimental data that make it possible to explain and predict the observed variability of turbulent-energy dissipation in the upper ocean are discussed. For this purpose, the dependence of the energy dissipation rate of breaking wind waves on their propagation velocity (see [1]) is used. The turbulent-energy dissipation values obtained earlier in [2, 3] by a direct method are compared to the results of radar measurements of individual breaking events presented in [1]. On the basis of this comparison, a strong dependence of the turbulent-energy dissipation value on the stage of wind-wave development, which is characterized by the ratio U _a /c _p (U _a is the wind speed and c _p is the phase speed of the peak of the wind-wave spectrum) is confirmed. This dependence was found earlier purely empirically. Moreover, it is shown that the theoretically obtained dependence (c _p /U _a )⁴, does not contradict the available empirical data. The results of this study opens possibilities for scientifically substantiated calculations of greenhouse-gas exchange (specifically, CO₂ exchange between the ocean and the atmosphere).     

Molecular phylogeny of the genus <Emphasis Type="Italic">Chondracanthus</Emphasis> (Rhodophyta), focusing on the resurrection of <Emphasis Type="Italic">C. okamurae</Emphasis> and the description of <Emphasis Type="Italic">C. cincinnus</Emphasis> sp. nov.

Determining the taxonomic status of the red algal genus Chondracanthus based on morphological characters is challenging due to the similarity and high degree of plasticity of the thallus. Since the taxonomic history of several Chondracanthus species remains unclear, we analyzed the plastid rbcL and mitochondrial COI genes of the specimens from Korea and Japan, in combination with morphological observations, to examine their phylogenetic relationships. Our results confirmed the distinction of C. okamurae, which is separated from C. intermedius, and identified a novel species, C. cincinnus sp. nov. Three species (C. okamurae, C. intermedius and C. cincinnus) formed a monophyletic clade with C. tenellus. C. okamurae is distinguished by linear, narrow, cylindrical to compressed, slightly recurved axes, and a high-intertidal to subtidal distribution. It was collected from Korea and Japan, while C. intermedius was identified from Japan only. A new species, Chondracanthus cincinnus sp. nov., is characterized by linear, compressed, strongly recurved axes, and a low-intertidal to subtidal distiribution. Based on the molecular phylogeny using rbcL and COI data, we herein resurrect C. okamurae as a distinct species and identify C. cincinnus as a new species.     

Changes in the egg production rate of two copepod species of the genus <Emphasis Type="Italic">Pseudocalanus</Emphasis> in relation to temperature in the White Sea

This study examines changes in egg production rates of two co-occurring Pseudocalanus species, P. acuspes and P. minutus, in response to changing temperatures (0, 3, 6, 9 and 12°C) in the White Sea. The boreal P. acuspes (sample size >650 individuals) increased its reproductive rate gradually across the entire range of temperatures from 0 to 12°C. Significant differences in egg production rates and reaction to temperature were also observed between P. acuspes females incubated during the early vs. the mid-summer season. The sample size of the Arctic P. minutus was significantly smaller due to the low numbers at which this species occurred. However, the results suggest that this species increases its egg production rates from 0 to 3°C from 3.3 to 8.5% dry weight female^–1 day^–1, but at 9° its reproductive rate drops significantly and the animals demonstrate markedly reduced fitness. Our results indicate that closely related co-occurring species, and even different generations within one species, can demonstrate significantly different responses to changes in the physical environment.     

Blooms of <Emphasis Type="Italic">Trichodesmium erythraeum</Emphasis> in the South Eastern Arabian Sea during the onset of 2009 summer monsoon

This study presents in situ evidence for the blooms of Trichodesmium erythraeum observed in the shelf waters of the South Eastern Arabian Sea (SEAS) during the onset of the southwest monsoon in June 2009. Evidence showed that water surface discoloration was caused by the accumulation of T. erythraeum, and that the water column contained a colony of T. thiebautii. The surface water color in the bloom region varied from pale brown to pinkish red. Pale brown indicated healthy algae at the peak of its photosynthetic activity, while pinkish red indicated the presence of photosynthetically less active filaments. Zooplankton abundance, especially copepodites, in the bloom area substantiated the theory that Trichodesmium filaments are excellent epiphytes to which the copepodites cling. The bloom area was very fertile with copious quantities of dissolved oxygen (6.85 ml L^?1), PO₄-P (0.108 μmol L^?1) and SiO₄ (1.29 μmol L^?1). Lower NO₃-N (0.028 μmol L^?1) values in the bloom area did not appear to affect Trichodesmium growth from molecular nitrogen fixation. However, lower NO₃-N values altered the normal phytoplankton composition of this area.     

Effect of Mercury Chloride on the Chlorophyl <Emphasis Type="Italic">a</Emphasis> and Pheophytin Content in Marine Microalgae: Measuring the Flow of Autotrophic Phytoplankton Using Sediment Traps Data

The present work aimed to study the influence of mercuric chloride (HgCl₂) on pigment content (chlorophyll a and pheophytin) in sediment traps on the basis of experimental data. The marine microalgal cultures of Conticribra weissflogii (Mediophyceae), Tetraselmis viridis (Chlorodendrophyceae), Amphidinium carterae (Dinophyceae), Nannochloropsis sp. (Eustigmatophyceae) were used. The obtained results revealed that the influence of mercury on pigment content as well as on photosynthetic activity was different in different species. The considerable decrease in phytopigments content was observed in Amphidinium carterae and Conticribra weissflogii, while the proportion of pheophytin increased. The pigment content of Tetraselmis viridis was not different from the control line, while for Nannochloropsis sp. the increase in pigment concentration was noticed. The complete immediate inhibition of photosynthetic activity was observed in Conticribra weissflogii, Tetraselmis viridis, Amphidinium carterae, while Nannochloropsis sp. responded only after 24 h of exposition. The pigment degradation rate was almost the same for diatoms and dinoflagellates. The equation for estimating the phytopigments content in sediment traps, that takes into consideration the exposition time, is proposed.     

Hybrid <Emphasis Type="Italic">N</Emphasis>-order Lagrangian interpolation Eulerian-Lagrangian method for salinity calculation

The Eulerian-Lagrangian method (ELM) has been used by many ocean models as the solution of the advection equation, but the numerical error caused by interpolation imposes restriction on its accuracy. In the present study, hybrid N-order Lagrangian interpolation ELM (LiELM) is put forward in which the N-order Lagrangian interpolation is used at first, then the lower order Lagrangian interpolation is applied in the points where the interpolation results are abnormally higher or lower. The calculation results of a step-shaped salinity advection model are analyzed, which show that higher order (N=3-8) LiELM can reduce the mean numerical error of salinity calculation, but the numerical oscillation error is still significant. Even number order LiELM makes larger numerical oscillation error than its adjacent odd number order LiELM. Hybrid N-order LiELM can remove numerical oscillation, and it significantly reduces the mean numerical error when N is even and the current is in fixed direction, while it makes less effect on mean numerical error when N is odd or the current direction changes periodically. Hybrid odd number order LiELM makes less mean numerical error than its adjacent even number order LiELM when the current is in the fixed direction, while the mean numerical error decreases as N increases when the current direction changes periodically, so odd number of N may be better for application. Among various types of Hybrid N-order LiELM, the scheme reducing N-order directly to 1st-order may be the optimal for synthetic selection of accuracy and computational efficiency.     

New records of three <Emphasis Type="Italic">Paraprionospio</Emphasis> species (Polychaeta: Spionidae) from Korean waters

The spionid polychaete Paraprionospio pinnata (Ehlers 1901) has been widely reported from Korean waters. We examined some specimens belonging to the genus Paraprionospio that had been collected from Korean waters, and identified them as Paraprionospio coora Wilson, 1990, Paraprionospio cordifolia Yokoyama, 2007 and Paraprionospio patiens Yokoyama, 2007, which are new to the fauna of Korea. The present study suggests misidentification of the specimens that have been previously reported as P. pinnata from Korean waters.     

Comparison of carbonate parameters and air–sea CO<Subscript>2</Subscript> flux in the southern Yellow Sea and East China Sea during spring and summer of 2011

In this work, we examined the carbonate parameters, i.e. total alkalinity (TA), pH, and partial pressure of CO₂ (pCO₂), and the air–sea CO₂ flux (FCO₂) in the continental shelves of the southern Yellow Sea (SYS) and East China Sea (ECS), based on two field surveys conducted in April and August of 2011. Surface pCO₂ showed significant spatial variations, ranging from 246 to 686 µatm in spring (average ± standard deviation = 379 ± 95 µatm) and from 178 to 680 µatm in summer (384 ± 114 µatm). During the spring cruise, the central SYS (pCO₂ < 240 µatm) and the Changjiang estuary (pCO₂ < 300 µatm) were under-saturated with CO₂, while the southern SYS and the southwestern ECS were supersaturated (pCO₂ = 420–680 µatm). In summer, however, the CO₂-supersaturated waters (pCO₂ = 380–680 µatm) occupied a relatively wide area, including the nearshore of the SYS and the Changjiang estuary, whereas pCO₂-deficient water (pCO₂ = 220–380 µatm) was observed only at the offshore ECS. In general, the entire SYS and ECS area behaved as a sustained CO₂ sink, with average FCO₂ of ?3.9 and ?2.1 mmol m^?2 d^?1 in spring and summer, respectively. Phytoplankton production was the driving force for CO₂ absorption, especially during the spring cruise. In addition, we found that typical water mixing processes and decomposition of terrestrial material were responsible for the release of CO₂ in three turbidity maximum regions.     

Effects of seawater acidification by ocean CO<Subscript>2</Subscript> sequestration on bathypelagic prokaryote activities

We investigated the effects of seawater acidification induced by ocean CO₂ sequestration on bathypelagic prokaryotes. We simulated acidification conditions by bubbling high-CO₂ air or adding chemical buffer solutions to seawater samples in order to examine changes in total cell counts, heterotrophic production rate, direct viable cell count, and relative abundance of Bacteria and Archaea. Considerable suppression of prokaryotic activities was observed at pH 7.0 or lower, especially in samples enriched with organic matter. The relative abundance of Archaea increased with increasing CO₂ concentration. We found that seawater acidification can potentially alter heterotrophic activities and community structure of bathypelagic prokaryotes.     

Phylogenetic relationship between symbionts of tubeworm <Emphasis Type="Italic">Lamellibrachia satsuma</Emphasis> and the sediment microbial community in Kagoshima Bay

Vestimentiferan tubeworms acquire their symbionts through horizontal transmission from the surrounding environment. In the present study, we constructed a 16S rRNA gene clone library to investigate the phylogenetic relationship between diverse microbes in the sediment and symbiotic bacteria in the trophosome of the tubeworm, Lamellibrachia satsuma, from Kagoshima Bay, Japan. Two symbiotic bacterial phylotypes belonging to the classes γ- and ε-Proteobacteria were found from this tubeworm trophosome. They were very closely related to the symbionts of several other marine invertebrates. The most predominant bacteria in the sediment were ε-Proteobacteria. A broad diversity of bacteria belonged to non-proteobacterial phyla such as Planctomycetes, Acidobacteria, and Chloroflexi was observed. The presence of sulfur oxidizers (i.e., ε-Proteobacteria and γ-Proteobacteria) and sulfur reducers (i.e., δ-Proteobacteria) may play a significant role in the sulfur cycle in these habitats and provide multiple sources of nutrition to the cold-seep communities. Closely related clones of ε-Proteobacteria symbiont in the species level and of γ-Proteobacteria symbiont in the genus level were found in the surrounding sediment. The similarity of symbiont clones of L. satsuma with other symbionts and free-living bacteria suggests the possibility of opportunistic symbiosis in ε-Proteobacteria and the co-evolution of γ-Proteobacteria having occurred after symbiosis with the tubeworms.