Spatial distribution, diversity and composition of bacterial communities in sub-seafloor fluids at a deep-sea hydrothermal field of the Suiyo Seamount

Spatial distribution, diversity, and composition of bacterial communities within the shallow sub-seafloor at the deep-sea hydrothermal field of the Suiyo Seamount, Izu-Bonin Arc, Western Pacific Ocean, were investigated. Fluids were sampled from four boreholes in this area. Each borehole was located near or away from active vents, the distance ranging 2–40 m from active vents. In addition, fluids discharging from a natural vent and ambient seawater were sampled in this area. We extracted DNA from each sample, amplified bacterial 16S rRNA genes by PCR, cloned the PCR products and sequenced. The total number of clones analyzed was 348. Most of the detected phylotypes were affiliated with the phylum Proteobacteria, of which the detection frequency in each clone library ranged from 84.6% to 100%. The bacterial community diversity and composition were different between hydrothermal fluids and seawater, between fluids from the boreholes and the vent, and even among fluids from each borehole. The relative abundances of the phylotypes related to Thiomicrospira, Methylobacterium and Sphingomonas were significantly different among fluids from each borehole. The phylotypes related to Thiomicrospira and Alcanivorax were detected in all of the boreholes and vent samples. Our findings provide insights into bacterial communities in the shallow sub-seafloor environments at active deep-sea hydrothermal vent fields.     

Spatial distribution of sister species of vesicomyid bivalves Calyptogena okutanii and Calyptogena soyoae along an environmental gradient in chemosynthetic biological communities in Japan

Vesicomyid bivalves have a substantial biomass in deep-sea chemosynthetic biological communities in the Pacific. Using a novel multiplex-PCR (mPCR) method to identify the co-occurring vesicomyids in Sagami Bay, we analyzed the distribution of Calyptogena okutanii and Calyptogena soyoae along environmental gradients. All the known distributions of C. okutanii indicated the different preferences in salinity and temperature to those of C. soyoae, and in Sagami Bay, depth seemed to be an important environmental factor, too. Although the concentration of hydrogen sulfide in sediment was not examined, our results showed that the distributions of these two Calyptogena clams were affected by salinity and temperature.     

Larval development ofEuphausia similis (Crustacea,Euphausiacea) in Sagami Bay,central Japan

The larvae ofEuphausia similis G. O. Sars in Sagami Bay, Central Japan, are described. Nauplius, metanauplius, calyptopis and furcilia stages are included. In the furcilia stage, six forms are identified on the basis of the form of the pleopods and the number of terminal telson spines. Furcilia I: a pair of non-setose pleopods and seven terminal telson spines. Furcilia II: a pair of setose and three pairs of non-setose pleopods and seven terminal spines. Furcilia III: four pairs of setose and a pair of non-setose pleopods and seven terminal spines. Furcilia IV: five pairs of setose pleopods and five terminal telson spines. Furcilia V: five pairs of setose pleopods and three terminal telson spines. Furcilia VI: five pairs of setose pleopods and one terminal telson spine. On the basis of the developmental pathway of the larvae,E. similis is suggested to be related toE. spinifera, E. longirostris, E. hanseni and theE. gibboides group species. The size ofE. similis larvae, expressed as total length, in Sagami Bay varied according to month. The sizes of calyptopis III and furcilia I–V stages were smallest in November and March and largest in May.     

Stage-V copepodites of Calanus sinicus and Calanus jashnovi (Copepoda: Calanoida) in mesopelagic zone of Sagami Bay as identified with genetic markers, with special reference to their vertical distribution

We applied genetic makers to identify Calanus species occurring in Sagami Bay, Japan, in order to investigate their vertical distribution in the upper 1000 m. First, interspecific genetic distances of three gene loci, mitochondrial small ribosomal RNA (srRNA), nuclear internal transcribed spacers 1 (ITS1) and 2 (ITS2), were estimated from morphologically distinguishable adult females of Calanus sinicus, Calanus jashnovi and Calanus pacificus that were collected from Sagami Bay, the Kuroshio Extension and the Oyashio region, respectively. The highest levels of interspecific genetic distance were observed in srRNA, followed by ITS1 and ITS2. The intraspecific genetic distances within C. sinicus were much lower than the interspecific genetic distances, indicating that DNA sequences in these loci are consistent with the morphological differences. This information was used as a criterion for species identification based on DNA sequence variation, and allowed us to identify the fifth copepodites (CVs) or younger stages of these species. Next, the vertical distribution of Calanus species was investigated in Sagami Bay in May 2006, on the basis of a stratified sampling in the upper 1000 m. By applying the genetic markers, 23 individuals comprising all copepodite stages were allocated into either C. sinicus or C. jashnovi, and the small- and large-sized CVs were identified as C. sinicus and C. jashnovi, respectively. The total abundance of C. sinicus was highest at 0-50 m and decreased with depth. On the contrary, CV individuals of C. sinicus were abundant not only in 0-50 m but also below 200 m with minimum occurrences in 150-200 m depth. C. jashnovi was much less abundant than C. sinicus and comprised of only CIV and CV which occurred in the upper 100 m and deeper than 50 m depths, respectively. The abundance of C. sinicus in the 1000-m water column of Sagami Bay was at a level comparable to that in shelf waters, suggesting the importance of off-shelf individuals in the biological production and organic transport in the respective areas.     

Comparing management of recreational Pagrus fisheries in Shark Bay (Australia) and Sagami Bay (Japan): Conventional catch controls versus stock enhancement

This paper compares the management of recreational fisheries for pink snapper (Pagrus auratus) in the inner gulfs of Shark Bay (Australia) and the closely related red sea bream (Pagrus major) in Sagami Bay (Japan). Fishing and other factors have resulted in population declines of these species in both regions. In response, fishery managers have employed contrasting management, more conventional catch controls in Shark Bay and stock enhancement in Sagami Bay. Although recreational harvest levels were higher than commercial levels in both fisheries, the driving mechanisms are comparatively different due to historical, social, economic and political issues in the respective locations.     

Microbial community structure and diversity in deep-sea hydrothermal vent sediments along the Eastern Lau Spreading Centre

The aim of this study is to investigate microbial structures and diversities in five active hydrothermal fields’ sediments along the Eastern Lau Spreading Centre (ELSC) in the Lau Basin (southwest Pacific). Microbial communities were surveyed by denatured gradient gel electrophoresis (DGGE) and clone library analysis of 16S rRNA genes. The differences in microbial community structures among sediment samples from the five deep-sea hydrothermal sites were revealed by DGGE profiles. Cluster analysis of DGGE profiles separated the five hydrothermal samples into two groups. Four different 16S rRNA gene clone libraries, representing two selected hydrothermal samples (19-4TVG8 and 19-4TVG11), were constructed. Twenty-three and 32 phylotypes were identified from 166 and 160 bacterial clones respectively, including Proteobacteria, Bacteroidetes, Firmicutes, Nitrospirae and Planctomycetes. The phylum Proteobacteria is dominant in both bacterial libraries with a predominance of Gamma-Proteobacteria. A total of 31 and 25 phylotypes were obtained from 160 and 130 archaeal clones respectively, including Miscellaneous Crenarchaeotic Group, Marine Group Ⅰ and Ⅲ, Marine Benthic Group E, Terrestrial Hot Spring Crenarchaeota and Deep-sea Hydrothermal Vent Euryarchaeota. These results show a variety of clones related to those involved in sulfur cycling, suggesting that the cycling and utilization of sulfur compounds may extensively occur in the Lau Basin deep-sea hydrothermal ecosystem.     

Diatom quantification and their distribution with salinity brines in coastal sediments of Terra Nova Bay (Antarctica)

Benthic diatoms represent an important element of global nutritional productivity; to raise attention on their role, which is often neglected due to analytical difficulties, surface (1 cm top layer) coastal sediments from Gerlache inlet to Penguin Bay at Terra Nova Bay were collected and stored at -20 °C. DNA amplification by real-time PCR, based on diatom-specific oligonucleotide primers designed on small-subunit rRNA (SSU rRNA), was performed in addition to diatom conventional cell counting and spectrophotometric determination of photo-pigments. Moreover, cations and anions were determined in sediments with the aim to identify factors involved in the control of diatom abundance. Diatom distribution was found quite heterogeneous displaying significant differences from site to site. The salinity in sediments ranged from 45.1 at Gerlache inlet to 76.2 at Penguin Bay and it was correlated with cell abundance, biodiversity, amount of pigments and amplified DNA. The dominant species, Fragilariopsis curta, was associated to sediment salinity brines.     

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.     

Characteristics of the eddy caused by Izu-Oshima Island and the Kuroshio branch current in Sagami Bay,Japan

Direct current measurements of the branch current of the Kuroshio intruding into Sagani Bay were carried out during 1989–1990 in order to clarify the frequency characteristics of the eddies in the lee of Izu-Oshima Island, which are well recognized as cold water mass produced by upwelling. Satellite and ADCP (Acoustic Doppler Current Profiler) data indicated that current velocity in the eddy fluctuates with periods of 2–4 days and 6–8 days.When the Kuroshio branch current intruding into Sagami Bay from the western channel is weak and its velocity at the depth of 400 m is approximately 10 cm s^–1, the 6–8 day period fluctuation is dominant. On the other hand, when the branch current strongly intrudes from the western channel with a velocity of approximately 20 cm s^–1, the 2–4 day period fluctuation dominates. The relationship between the periods and velocities agrees well with theory based on laboratory experiments for a flow of a homogeneous fluid past a circular obstacle. These periods correspond to the time scale of appearance of the eddy caused by the intrusion of the Kuroshio branch current into Sagami Bay and Izu-Oshima Island.     

Growth and succession patterns of major phylogenetic groups of marine bacteria during a mesocosm diatom bloom

Our objective was to track microbial processes associated with serial degradation of organic matter derived from algal blooms. To do this, we analyzed population fluctuations and growth responses of major phylogenetic groups of free-living marine bacteria. We used bromodeoxyuridine immunocytochemistry–fluorescence in situ hybridization methodology to examine marine bacterial community development during and after a diatom bloom in a mesocosm. We revealed that the Roseobacter/Rhodobacter, SAR11, Alteromonas, and Bacteroidetes groups were clearly major phylotypes responsible for most free-living bacterial biomass and production throughout the experiment. The clearest bacterial response was a proliferation of the Alteromonas group (cells with large volumes) during development of the bloom (up to 30?% of actively growing cells). Populations of these bacteria declined sharply thereafter, likely due to grazing. Alteromonas group responses suggest that these bacteria strongly influenced the flux of organic matter at an early bloom stage. The growth potential of Bacteroidetes was relatively large as the bloom peaked; this early development probably contributed to the initial stage of bloom decomposition. In contrast, the contribution of Roseobacter/Rhodobacter to total bacterial production increased at a late stage of decomposing of the bloom. The contributions of Betaproteobacteria, SAR11, and SAR86 groups to total bacterial abundance and production were relatively minor throughout the experiment. These results imply that the ability to utilize organic matter derived from diatoms varies among bacterial phylotypes, and, frequently, less abundant but ecological specialist taxa such as Alteromonas may play major roles in the flux of organic matter during diatom blooms.     

Environmental changes in the Amur Bay (Japan/East Sea) during the last 150 years revealed by examination of diatoms and silicoflagellates

Siliceous unicellular microalgae — diatoms and silicoflagellates from sediments in Amur Bay were analyzed with high temporal resolution to examine changes over the last 150 years. The age of sediments was estimated from unsupported ²¹⁰Pb controlled by ¹³⁷Cs. Siliceous microalgae examined in each cm of two sediment cores demonstrated significant changes in the ecological structure of the assemblages that reflected changes in sedimentation conditions. During the years 1860–1910 the sediments accumulated under the great influence of river runoff. For about the next 50 years the number of freshwater species and marine benthic diatoms in sediments sharply declined, which is probably connected with the weakening of the effects of river runoff due to deforestation. Since the early 1960s the sedimentation conditions in the Amur Bay changed significantly. Marine planktonic diatoms and silicoflagellates began to prevail in sediments and this reflects increasing microphytoplankton productivity. One consequence of this was the formation of seasonal bottom hypoxia in Amur Bay. The ecological structure of diatom and silicoflagellate assemblages indicates that the sea level began to rise since the early 1960s and this corresponds to the water and air temperature increase in the area for that period. The obtained data suggest that the environmental changes over the last 150 years in Armur Bay are associated with the weakening of river runoff due to deforestation, sea level rise caused by global warming, and the increase of siliceous microplankton productivity that resulted in the formation of seasonal bottom hypoxia.

     

西南印度洋深海热液硫化物区沉积物微生物群落结构和固氮基因多样性

A sediment sample was collected from a deep-sea hydrothermal vent field located at a depth of 2 951 m on the Southwest Indian Ridge. Phylogenetic analyses were performed on the prokaryotic community using polymerase chain reaction（PCR） amplification of the 16 S rRNA and nifH genes. Within the Archaea, the dominant clones were from marine benthic group E（MBGE） and marine group I（MGI） belonging to the phyla Euryarchaeota and Thaumarchaeota, respectively. More than half of the bacterial clones belonged to the Proteobacteria, and most fell within the Gammaproteobacteria. No epsilonproteobacterial sequence was observed. Additional phyla were detected including the Actinobacteria, Bacteroidetes, Planctomycetes, Acidobacteria, Nitrospirae, Chloroflexi, Chlorobi, Chlamydiae, Verrucomicrobia, and candidate divisions OD1, OP11, WS3 and TM6, confirming their existence in hydrothermal vent environments. The detection of nifH gene suggests that biological nitrogen fixation may occur in the hydrothermal vent field of the Southwest Indian Ridge. Phylogenetic analysis indicated that only Clusters I and III NifH were present. This is consistent with the phylogenetic analysis of the microbial 16 S rRNA genes, indicating that Bacteria play the main role in nitrogen fixation in this hydrothermal vent environment.     

Seasonal patterns of growth and composition of phytoplankton in the lower Chesapeake Bay and vicinity

A twenty-three month study of lower Chesapeake Bay phytoplankton was made from February 1982 to December 1983. The major seasonal growth periods were dominated by a diatomaceous flora and a pico-nanoplankton complex composed mainly of cyanobacteria, chlorophytes and other cells <5μm in size. There was a pattern of multiple pulses throughout the year. However, the trend for maximum development occurred during the winter-spring and fall periods. The dominant diatoms were Skeletonema costatum, Leptocylindrus danicus, and Asterionella glacialis.Seasonal comparisons were made to earlier phytoplankton composition studies in the Bay. Over the past 60 years there has been a change in phytoplankton assemblages and concentrations in the Bay. Skeletonema costatum has remained a dominant species, but a more abundant and broader base of small sized, chainforming diatoms have become established. In addition, high concentrations of the pico-nanoplankton, chrysophyceans and cryptophyceans were abundant.     

Lagrangian observation of phytoplankton dynamics at an artificially enriched subsurface water in Sagami Bay,Japan

Phytoplankton dynamics in the lower euphotic zone were observed by tracking a subsurface water released at 20-m depth from Takumi, an artificial upwelling device. Takumi continually discharged seawater pumped up from a depth of 205 m: this water was mixed with 5-m depth water to adjust the density to that of 20-m depth water of Sagami Bay, Japan. The discharged water was pulse-labeled at Takumi with uranine and tracked for 63.9 h with a drifting buoy equipped with a drogue at 20-m depth. We present a simple model to estimate in situ phytoplankton net growth rates from temporal changes in phytoplankton abundance in the discharged water with correction for the influence of water exchange between the discharged water and neighboring layers. Lagrangian observation showed active growth of pico- and nanophytoplankton, especially cryptophytes and Synechococcus (Cyanobacteria), in the subsurface layer. In contrast, diatoms grew little in spite of micromolar concentrations of nutrients. The active growth of pico- and nanophytoplankton was in good agreement with shipboard serial dilution culture experiments. The low growth activity of diatoms was suggested to be related to low light availability in the subsurface layer.     

Difference in the prevailing periods of internal tides between Sagami and Suruga Bays: Numerical experiments

Current measurements in the surface layer in Sagami and Suruga Bays showed existence of significant tidal currents which are considered to be mainly due to internal tides (Inaba, 1982; Ohwaki,ea al., 1991). In addition, the prevailing period of the tidal currents is semidiurnal in Sagami Bay, but diurnal in Suruga Bay. To explain this difference in the prevailing, periods, numerical experiments were carried out using a two layer model. The internal tides are generated on the Izu Ridge outside the two bays. The semidiurnal internal tide propagates into Sagami Bay having characteristics of an internal inertia-gravity wave, while it propagates into Suruga Bay having characteristics of either an internal inertia-gravity wave or an internal Kelvin wave. The diurnal internal tide behaves only as an internal Kelvin wave, because the diurnal period is longer than the inertia period. Thus, the diurnal internal tide generated on the Izu Ridge can be propagated into Suruga Bay, while it cannot propagate into the inner region of Sagami Bay, though it is trapped around Oshima Island, which is located at the mouth of Sagami Bay. The difference in the propagation characteristics between the semidiurnal and diurnal internal tides can give a mechanism to explain the difference in the prevailing periods of the internal tides between Sagami and Suruga Bays.     

利用扩增vvhA基因的环介导等温扩增技术快速，特异而敏感地检测创伤弧菌

创伤弧菌是一种可感染人类的河口病原菌。建立快速,特异而敏感的检测方法,有助于创伤弧菌感染的早期疾病诊断和及时治疗。本研究设计了针对vvhA基因的一系列引物（包括两对外部引物和两对内部引物）,采用环介导等温扩增技术（LAMP）来检测创伤弧菌。结果显示,本方法的最适扩增温度是63℃,反应仅需35分钟。扩增产物不仅可以用含有DNA ladder的琼脂糖凝胶电泳检出,也可借助钙黄绿素直接肉眼观察。采用45株菌株检测该方法的特异性,其中所有创伤弧菌均被检出而其他菌株检测结果皆为阴性。本方法的敏感性是普通PCR扩增的100倍,同时利用该方法可以准确检测出所有的模拟样品、临床标本及环境样品。与其他已知方法比较,针对vvhA基因的介导等温扩增技术可以快速、简单、敏感及特异地鉴定创伤弧菌。     

Evidence for predominance of internal tidal currents in Sagami and Suruga bays

Tidal currents observed in a surface layer overlying deep water in Sagami and Suruga Bays frequently have large amplitude in summer and fall. Numerical experiments show that the current amplitude due to the surface tides is below 1.0 cm sec^–1 for the semidiurnal and diurnal constituents in the inner region of the two bays. The observed current amplitudes are larger than the calculated ones due to the surface tides. Therefore, the observed tidal currents are indicated to be due mainly to the internal tides. In addition, the semidiurnal currents dominate the diurnal currents in Sagami Bay, while the opposite occurs in Suruga Bay. These results suggest that the prevailing periods of the internal tides differ between the two bays,i.e., the internal tide has a semidiurnal period in Sagami Bay and a diurnal period in Suruga Bay.     

Intrusion events of the intermediate Oyashio Water into Sagami Bay, Japan

Intermediate intrusion of low salinity water (LSW) into Sagami Bay was investigated on the basis of CTD data taken in Sagami Bay and off the Boso Peninsula in 1993–1994. In October 1993, water of low temperature (<7.0°C), low salinity (<34.20 psu) and high dissolved oxygen concentration (>3.5 ml I⁻¹) intruded along the isopycnal surface of {ie29-1} at depths of 320–500 m from the Oshima East Channel to the center of the bay. On the other hand, the LSW was absent in Sagami Bay in the period of September–November 1994, though it was always found to the south off the Boso Peninsula. Salinity and dissolved oxygen distributions on relevant isopycnal surfaces and water characteristics of LSW cores revealed that the LSW intruded from the south off the Boso Peninsula to Sagami Bay through the Oshima East Channel. The LSW cores were distributed on the continental slope along 500–1000 m isobaths and its onshore-offshore scales were two to three times the internal deformation radius. Initial phosphate concentrations in the LSW revealed its origin in the northern seas. These facts suggest that the observed LSW is the submerged Oyashio Water and it flows southwestward along the continental slope as a density current in the rotating fluid. The variation of the LSW near the center of Sagami Bay is closely related to the Kuroshio flow path. The duration of LSW in Sagami Bay is 0.5 to 1.5 months.     

PCB resistance within phytoplankton populations in polluted and unpolluted marine environments

A comparison between clones of two species of diatoms, Asterionella japonica and Ditylum brightwellii, isolated from Sandy Hook, New Jersey (polluted site) and Montauk, New York (unpolluted site) indicated that some of the clones from Sandy Hook were more resistant to polychlorinated biphenyls (PCB). A japonica was less sensitive to PCB than D. brightwellii since seven clones of A. japonica from both sites were tolerant of 25 μg litre^?1 PCB whereas no clones from either site of D. brightwellii showed such resistance. Growth under high light intensity increased the sensitivity to PCB in all clones except one super-resistant clone from Sandy Hook which was not affected even by additions of 50 μg litre^?1 to the growth medium. This interactive effect of light on PCB sensitivity may relate to the photo-adaptive characteristics of each clone.