Growth characteristics of low-nutrient bacteria from the north-east and central Pacific Ocean

Abstract Low-nutrient, or oligotrophic, marine bacteria, defined operationally as those which grow well in the dilute nutrient concentrations typical of natural seawaters, were found in waters of the eastern and central Pacific Ocean. All isolates tested grew in sterile unsuplemented seawater, ultraviolet-irradiated seawater, artificial seawater and charcoal-treated artificial seawater. Under optimal conditions, doubling times for low-nutrient bacteria were generally 2–5 h. Low-nutrient bacteria are heterotrophs responding to added organic substrates including glucose, proline, acetate, peptone, algal extract and yeast extract. Organic substrate concentrations ( K _s values) necessary to yield half the maximum growth rates of low-nutrient bacteria ranged from 0.8–12 μg · 1⁻¹ (0.2–4.9 μg C · 1⁻¹). Bacterial inoculum size had little effect on doubling times, whereas low oxygen tensions and low temperatures reduced growth rates. Surfaces (glass beads) increased growth rates and maximum cell densities. The results of these laboratory studies indicate that low-nutrient bacteria play a major role in marine heterotrophic transformations of dissolved organic matter.     

Environmental fungal diversity in the upwelling ecosystem off central Chile and potential contribution to enzymatic hydrolysis of macromolecules in coastal ecotones

Even though occurrence of fungi in several marine environments has been documented, their inclusion within the marine microbial loop is not fully recognized. A major constraint is whether fungi in coastal waters are truly marine or represent transient microorganisms transported from terrestrial environments. We approached this issue by analyzing ambient fungal composition and hydrolytic activity of culturable fungi along a nearshore-offshore gradient in the upwelling ecosystem off central Chile, a region of high marine productivity strongly influenced by river discharges. We detected different communities of fungi in nearshore and offshore waters, with near estuary strains hydrolyzing proteins and carbohydrates faster than those from offshore sites. We conclude that coastal waters off central Chile comprise distinct fungal communities representative of offshore and nearshore environments, and provide new evidence for fungi processing organic matter in coastal ecotones, opening a fresh perspective for disappearance of organics carried by rivers in the coastal ocean.     

Advective Mixing of Pore Waters and Ecology of Sandy Sediments of the Ocean Shelf

Sand sediments are widely distributed in the shelf zone of the World Ocean. The main physical and ecological peculiarity of marine sands is the mobility of pore waters and their mixing with the near-bottom waters of the sea. The mixing is closely related to turbulent processes in the water column; the most important of these processes is wave mixing. This causes filtration of seawater through the sand, ensuring therefore, perhaps, the most large-scale filtration process on the Earth. Advective mixing redistributes dissolved oxygen and nutrients between the pore waters of the sand and the bulk of the seawater column and determines redox conditions in the sediment column, which usually are either oxidative or suboxic; the metals with variable valence, mostly iron, serve as a redox buffer. The regeneration of nutrients and nitrification of ammonia also take place in the sand column. The instability of advective mixing is considered as the major difficulty for qualitative assessment of energetic metabolism and regeneration of nutrients in sands as well as for revealing the ecological role of marine sands in coastal ecosystems.     

The distribution of major biochemical components in the waters of the Kuril Islands

In this paper, data are presented on the distribution of dissolved and particulate carbohydrates, proteins, lipids, and nucleic acids (NAs) in the waters of the Kuril Islands on both the Sea of Okhotsk and Pacific Ocean sides. By analyzing discrepancies in the absolute and relative concentrations of these substances, it is revealed that the waters of the Sea of Okhotsk have a strong bioproductivity. Changes in the absolute and relative conventrations of the major biochemical components of dissolved organic matter (DOM) and particulate organic matter (POM) are shown to result from the biological activity of auto- and heterotrophs of shelf and deep-sea ecosystems. Carbohydrates and proteins prove to be the major biochemical components of DOM and POM, respectively. Given high primary productivity, the carbohydrate content in POM is comparable to that of protein. In the southern Kuril area, the high concentration of dissolved and particulate NAs (which exceeds that of dissolved protein) and the high content of dissolved lipids are both strongly associated with local fish and zooplankton distribution. In the shelf and pelagic waters, biological factors dominate the horizontal and vertical distribution patterns of the major biochemical components of DOM and POM. The hydrological regime only affects these patterns in the straits of the Kuril Islands. For the first time, the biochemical composition of DOM and POM in the sea surface microlayer (SML) is characterized. In this microlayer, the concentrations of major biochemical components of DOM and POM are, to a great extent, higher than in the underlying layer.     

Distribution of thermophilic marine sulfate reducers in north sea oil field waters and oil reservoirs

The distribution of thermophilic marine sulfate reducers in produced oil reservoir waters from the Gullfaks oil field in the Norwegian sector of the North Sea was investigated by using enrichment cultures and genus-specific fluorescent antibodies produced against the genera Archaeoglobus, Desulfotomaculum, and Thermodesulforhabdus. The thermophilic marine sulfate reducers in this environment could mainly be classified as species belonging to the genera Archaeoglobus and Thermodesulforhabdus. In addition, some unidentified sulfate reducers were present. Culturable thermophilic Desulfotomaculum strains were not detected. Specific strains of thermophilic sulfate reducers inhabited different parts of the oil reservoir. No correlation between the duration of seawater injection and the numbers of thermophilic sulfate reducers in the produced waters was observed. Neither was there any correlation between the concentration of hydrogen sulfide and the numbers of thermophilic sulfate reducers. The results indicate that thermophilic and hyperthermophilic sulfate reducers are indigenous to North Sea oil field reservoirs and that they belong to a deep subterranean biosphere.     

Degradation of Ferric Chelate of Ethylenediaminetetraacetic Acid by Bacterium Isolated from Deep-Sea Stalked Barnacle

Twenty strains of marine bacteria that degrade ferric chelate of ethylenediaminetetraacetic acid (Fe-EDTA) were isolated from among 117 strains collected from a marine environment. Among them strain 02-N-2, which was isolated from stalked barnacle collected from the deep sea in the Indian Ocean, had the highest Fe-EDTA degradation ability and was selected for further study. The strain showed high Fe-EDTA degradation ability at different seawater concentrations. In addition, the intact cells of this strain had the ability to degrade such metal-EDTAs as Ca, Cu, and Mg. The strain was an aerobic, gram-variable, rod-shaped organism. The results of various taxonomic studies revealed that the strain had significant similarity to Bacillus jeotgali JCM 10885^T, which was isolated from a Korean traditional fermented seafood, Jeotgal.     

Effect of melting Antarctic sea ice on the fate of microbial communities studied in microcosms

Although algal growth in the iron-deficient Southern Ocean surface waters is generally low, there is considerable evidence that winter sea ice contains high amounts of iron and organic matter leading to ice-edge blooms during austral spring. We used field observations and ship-based microcosm experiments to study the effect of the seeding by sea ice microorganisms, and the fertilization by organic matter and iron on the planktonic community at the onset of spring/summer in the Weddell Sea. Pack ice was a major source of autotrophs resulting in a ninefold to 27-fold increase in the sea ice-fertilized seawater microcosm compared to the ice-free seawater microcosm. However, heterotrophs were released in lower numbers (only a 2- to 6-fold increase). Pack ice was also an important source of dissolved organic matter for the planktonic community. Small algae (<10 μm) and bacteria released from melting sea ice were able to thrive in seawater. Field observations show that the supply of iron from melting sea ice had occurred well before our arrival onsite, and the supply of iron to the microcosms was therefore low. We finally ran a “sequential melting” experiment to monitor the release of ice constituents in seawater. Brine drainage occurred first and was associated with the release of dissolved elements (salts, dissolved organic carbon and dissolved iron). Particulate organic carbon and particulate iron were released with low-salinity waters at a later stage.     

A note on the isolation and enumeration of Gram positive cocci from marine and estuarine waters

Ninety-two percent of Gram positive cocci detected in estuarine and marine surface waters were identified as Staphylococcus spp. Micrococcus and Streptococcus spp. were rarely isolated from seawater, except at dump site stations located in the Puerto Rico Trench area of the Atlantic Ocean, where they were present in unusually large numbers. Staphylococci were present in greatest abundance near land, but were also found in open ocean surface waters in low, but relatively constant, numbers. Staphylococcus epidermidis , or strains related to Staph. hominis , were present in all samples. Staphylococci from surface water samples collected at the Puerto Rico Trench dump site degraded a wide range of compounds. It is considered that these cocci were derived from the wastes dumped, as very few were found in unpolluted marine waters and in those cases in which stimulatory factors, such as disposal of wastes, do not play a role, Gram positive cocci will not be recovered unless large volumes of water are filtered. The Gram positive cocci are present in very low numbers in natural seawater of the open ocean and, therefore, are commonly overlooked. The significance of these bacteria in the nutrient cycles of the sea remains to be elucidated. Documentation of a widespread occurrence of Gram positive cocci in North Eastern Atlantic seawater is provided.     

Diversity,Ecology and Biogeochemistry of Cyst-Forming Acantharia (Radiolaria) in the Oceans

Marine planktonic organisms that undertake active vertical migrations over their life cycle are important contributors to downward particle flux in the oceans. Acantharia, globally distributed heterotrophic protists that are unique in building skeletons of celestite (strontium sulfate), can produce reproductive cysts covered by a heavy mineral shell that sink rapidly from surface to deep waters. We combined phylogenetic and biogeochemical analyses to explore the ecological and biogeochemical significance of this reproductive strategy. Phylogenetic analysis of the 18S and 28S rRNA genes of different cyst morphotypes collected in different oceans indicated that cyst-forming Acantharia belong to three early diverging and essentially non symbiotic clades from the orders Chaunacanthida and Holacanthida. Environmental high-throughput V9 tag sequences and clone libraries of the 18S rRNA showed that the three clades are widely distributed in the Indian, Atlantic and Pacific Oceans at different latitudes, but appear prominent in regions of higher primary productivity. Moreover, sequences of cyst-forming Acantharia were distributed evenly in both the photic and mesopelagic zone, a vertical distribution that we attribute to their life cycle where flagellated swarmers are released in deep waters from sinking cysts. Bathypelagic sediment traps in the subantarctic and oligotrophic subtropical Atlantic Ocean showed that downward flux of Acantharia was only large at high-latitudes and during a phytoplankton bloom. Their contribution to the total monthly particulate organic matter flux can represent up to 3%. High organic carbon export in cold waters would be a putative nutritional source for juveniles ascending in the water column. This study improves our understanding of the life cycle and biogeochemical contribution of Acantharia, and brings new insights into a remarkable reproductive strategy in marine protists.     

Impact of sinking carbon flux on accumulation of deep-ocean carbon in the Northern Indian Ocean

The export of carbon through the biological pump from the surface to the deep ocean has a direct influence on the removal of CO₂ from the atmosphere. This is because the carbon is sequestered for only a few days to months in the surface while the carbon removed from the surface to deep waters takes hundreds of years to re-enter the atmosphere. The highest dissolved inorganic carbon (DIC) is expected in the deep waters of the North Pacific due to longer age of waters. On contrary, the higher deep water DIC is found in the northern Indian Ocean than elsewhere in the World Oceans. The sinking fluxes of particulate organic (POC) and inorganic carbon (CaCO₃) are found to be the highest in the northern Indian Ocean. The rates of bacterial respiration, organic carbon regeneration and inorganic carbon dissolution are also found to be the highest in the northern Indian Ocean than elsewhere. A most efficient biological pump appears to be operating in the northern Indian Ocean that transports surface-derived organic/inorganic carbon to deeper layers where it is converted and stored for longer times in dissolved inorganic form.     

Distribution of Vibrio parahaemolyticus and Related Organisms in the Atlantic Ocean off South Carolina and Georgia

The distribution of Vibrio parahaemolyticus and related organisms in the Atlantic Ocean was determined during the summer of 1971 from samples collected at stations along four transects on the continental shelf off the South Carolina and Georgia coasts. No V. parahaemolyticus strains were isolated from any of the samples of seawater (surface and bottom), sediment, and plankton which were collected. A numerical taxonomy analysis of data on substrate utilization, including 154 organic compounds serving as single carbon sources, was carried out, and four groups of strains were observed. Each group showed well-separated distribution profiles from shore out to the continental shelf. That is, the groupings were observed to correspond to coastal, off-shore and intermediate distribution patterns for the strains. This study provides a useful example of the kind of ecological distributional analysis of bacteria which can be accomplished with numerical taxonomy.     

Characterization and Relatedness of Marine Vibrios Pathogenic to Fish: Physiology, Serology, and Epidemiology

Cultural characteristics and serological relationships of pathogenic marine vibrios isolated from fish in the Pacific Northwest were studied. These organisms were compared with cultures of Vibrio anguillarum, a known fish pathogen. On the basis of morphological and cultural characteristics, the Pacific Northwest strains of Vibrio were found to be closely related to V. anguillarum. Serological analyses of thermostable antigens served to distinguish three serotypes among the vibrios. Serotype 1 was composed of organisms isolated from Northwest salmonids; serotype 2 of strains of V. anguillarum from European waters; and serotype 3 of organisms isolated from Pacific herring. The epidemiology of vibrio disease among populations of fish in the Pacific Northwest is discussed.     

Tracing biogeochemical subsidies from glacier runoff into Alaska's coastal marine food webs

Nearly half of the freshwater discharge into the Gulf of Alaska originates from landscapes draining glacier runoff, but the influence of the influx of riverine organic matter on the trophodynamics of coastal marine food webs is not well understood. We quantified the ecological impact of riverine organic matter subsidies to glacier‐marine habitats by developing a multi‐trophic level Bayesian three‐isotope mixing model. We utilized large gradients in stable (δ¹³C, δ¹⁵N, δ²H) and radiogenic (Δ¹⁴C) isotopes that trace riverine and marine organic matter sources as they are passed from lower to higher trophic levels in glacial‐marine habitats. We also compared isotope ratios between glacial‐marine and more oceanic habitats. Based on isotopic measurements of potential baseline sources, ambient water and tissues of marine consumers, estimates of the riverine organic matter source contribution to upper trophic‐level species including fish and seabirds ranged from 12% to 44%. Variability in resource use among similar taxa corresponded to variation in species distribution and life histories. For example, riverine organic matter assimilation by the glacier‐nesting seabirds Kittlitz's murrelet (Brachyramphus brevirostris) was greater than that of the forest‐nesting marbled murrelet (B. marmoratus). The particulate and dissolved organic carbon in glacial runoff and near surface coastal waters was aged (12100–1500 years BP ¹⁴C‐age) but dissolved inorganic carbon and biota in coastal waters were young (530 years BP ¹⁴C‐age to modern). Thus terrestrial‐derived subsidies in marine food webs were primarily composed of young organic matter sources released from glacier ecosystems and their surrounding watersheds. Stable isotope compositions also revealed a divergence in food web structure between glacial‐marine and oceanic sites. This work demonstrates linkages between terrestrial and marine ecosystems, and facilitates a greater understanding of how climate‐driven changes in freshwater runoff have the potential to alter food web dynamics within coastal marine ecosystems in Alaska.     

Origin, biochemical composition and vertical flux of particulate organic matter under the pack ice in Terra Nova Bay (Ross Sea, Antarctica) during late summer 1995

Water samples and particulate materials settling under the pack ice were collected in an ice-covered area near the Terra Nova Bay Italian Station during late summer 1995, in order to study short-term changes in the biochemical composition of particulate organic matter. At the end of the study period the phytoplankton biomass increase (up to >3.0 μg chlorophyll-a l⁻¹) was probably related to the intrusion under the pack ice of chlorophylls-enriched surface waters coming from the near ice-free area. Such increase was associated also with a notable increase in particulate organic matter concentrations, as well as in particulate organic matter vertical fluxes (up to >100 mg C m⁻² day⁻¹). Proteins were the most abundant biochemical class of particulate organic matter (on average about 49%), followed by lipids (29%) and carbohydrates (22%). By contrast, organic matter collected in the sediment trap was characterized by the dominance of lipids (about 55% of the total biopolymeric carbon flux) over carbohydrates (28%) and proteins (17%). The hydrolizable particulate biopolymeric carbon accounted for about 23% of total biopolymeric carbon. This value was about one-half of that found in ice-free waters, suggesting that the suspended particulate organic material under the pack ice was less digestible than in ice-free waters or was already partially digested. Despite this, and the decay of labile organic compounds in the sediment trap during the deployment, material settling towards the sea bottom under the pack ice in Terra Nova Bay, owing to its high lipid content, might represent an important high-quality food source for benthic consumers. Finally, assuming as possible the intrusion under sea ice of primary organic matter-enriched waters, we hypothesize the occurrence of a “fertilization” effect deriving from ice-melting areas towards under-ice waters, supplying the latter with an additional rate of primary organic matter. Accepted: 18 February 1999     

The composition and distribution of the particulate matter in the Strait of Magellan (Chile) during the 1991 and 1995 Italian campaigns

During 1991 and 1995, the Italian National Program for Antarctic Research carried out two oceanographic campaigns in the framework of the International (Strait of) Magellan Project. In this paper, we describe the distribution, biochemical composition, and mineralogical characteristics of particulate matter and the characteristics of the water masses defining microbasins in the Strait of Magellan. The data analyses highlighted differences in quality and quantity of the suspended matter and its organic component in the basins that make up the Strait and the Pacific Ocean. The westernmost basin is subject to an eastward flow from the Pacific Ocean, and continental runoff in the surface layers, which are consequently rich in organic matter with a high C:N ratio. The central basin, Isla Carlos III–Segunda Angostura, is characterized by the mixing of Sub-Antarctic Pacific waters, continental runoff, and glacio-fluvial waters: The basin has the lowest particulate matter concentration, but at Paso Ancho showed high concentrations of organic matter with a lower C:N ratio. The easternmost basin, Segunda Angostura-Atlantic entrance, is characterized by the mixing of the water column due to strong Atlantic tides and showed high concentration of particulate matter with a high detritic component. Generally, the organic matter concentrations showed significant differences during the cruises, being higher in 1991. Using the multivariate discriminant factor analysis to determine whether statistically significant differences existed between defined sampling areas, we determined that temperature, particulate organic carbon, and total particulate matter were the variables most important for the difference between areas.     

印度洋可培养解有机磷细菌的多样性及解磷特性

【目的】筛选获得印度洋海水解有机磷细菌,研究其解有机磷作用机理,初步了解该地区可培养有机磷细菌的系统发育多样性。【方法】采用卵磷脂培养基从分离自印度洋的细菌中筛选解有机磷细菌,根据16S rRNA基因序列确定获得的有机磷细菌的分类地位;同时挑选解磷效果显著的3株细菌利用液体发酵进行产酶和解磷特性分析。【结果】自916株细菌中得到99株具有解有机磷活性的细菌,分属于16个属。在以卵磷脂为其有机磷来源时,菌株India-BSP-1 (Cobetia sp.)、India-BSP-21 (Pelagibaca sp.)、India-BSP-23 (Pelagibacterium sp.)的培养液中磷酸盐(DIP)浓度曲线为N型,并且碱性磷酸酶的检测活性滞后于DIP的生成。【结论】印度洋海水中解有机磷细菌种类多样性丰富,疑似有新种;解有机磷细菌的解磷特性受DIP和碱性磷酸酶相互作用的影响。     

Development of a heterotrophic bacterial community within a closed prawn aquaculture system

The quantitative and qualitative development of a heterotrophic bacterial community in seawater was studied throughout an experimental rearing of the prawnPenaeus japonicus. The maturation of juvenile prawns had been carried out for 8 months in aerated tanks of seawater without any water renewal. Bacteria (337 strains) were isolated from seawater, which had been sampled at different times. Samples from one of the mussels used to feed the prawns and from the digestive tract of one prawn each supplied 40 more strains. 101 tests were performed on each strain, and cluster analysis showed the existence of 4 different groups containing 95% of the sampled strains. Characteristics of the various samples and groups were described in terms of ecotype diversities, catabolic potentialities, nutritional capacities, and morpho-physiological groups. Until nitrification attained a steady state, the heterotrophic community clearly decreased in number and was composed mostly of auxotrophic bacteria (pseudomonads andMoraxella-Acinetobacter groups). These bacteria needed growth factors and were unable to use amino acids (group B). At the end of the experiment (7 months) the bacteria isolated from the water were vibrios and enterobacteria, as were those isolated from the prawn and the mussel. They clustered together in group A. The number of strains that clustered with those of natural seawater (group E) decreased steadily during the experimental period.     

Physiological and Biochemical Characteristics of the Halophilic Bacteria Vibrio parahaemolyticus and V. alginolyticus Isolated from Marine Invertebrates of Peter the Great Bay, Sea of Japan

One hundred strains of halophilic vibrios were isolated from 16 species of marine invertebrates of Peter the Great Bay. Based on their morphological and biochemical characteristics, the bacteria were identified as Vibrio parahaemolyticus and Vibrio alginolyticus. Bacterial isolates possessed virulence enzymes (DNAase, lecithinase, catalase) and were characterized by a high enterotoxigenicity. It was determined that 76% of the V. parahaemolyticus strains and 43% of the V. alginolyticus strains were Kanagawa-positive. The isolates showed a high adhesive capability, the average adhesion index was 18.06 cells per erythrocyte for V. parahaemolyticus and 12.55 for V. alginolyticus. The results of this study suggest a high pathogenic potential of the isolated halophilic vibrios, which are an epidemic hazard to marine invertebrates and to humans.     

Distribution and ecology of Vibrio vulnificus and other lactose-fermenting marine vibrios in coastal waters of the southeastern United States.

Water, sediment, plankton, and animal samples from five coastal sites from North Carolina to Georgia were sampled for their lactose-fermenting vibrio populations. Over 20% of all vibrios tested were sucrose negative and o-nitrophenyl-beta-D-galactopyranoside (ONPG) positive, suggesting identification as the human pathogen Vibrio vulnificus. These vibrios were isolated from all sample sites and sources (water, sediment, plankton, and animals). Correlations with several of 19 environmental parameters monitored at each site were found for total vibrios. The presence of ONPG-positive, sucrose-negative vibrios was correlated with hydrocarbon levels in the water and, in the case of plankton samples, with salinity. A total of 279 sucrose-negative, ONPG-positive isolates were subjected to numerical taxonomic analysis, which resulted in three major clusters. Cluster I corresponded to and included 11 reference strains of V. vulnificus. Cluster II contained the largest number (133) of isolates, of which the great majority were bioluminescent. Although having a resemblance to V. harveyi, the isolates were ONPG positive and many were H2S positive. Cluster III consisted of strains similar to the group F vibrios (V. fluvialis). Of all of the isolates, 55% were luminescent, of which over 20% were lethal when injected into mice. Problems involved in detecting lactose fermentation among marine vibrios and the potential pathogenicity of these organisms are discussed.     

Distribution and functions of TonB-dependent transporters in marine bacteria and environments: implications for dissolved organic matter utilization

Background

Bacteria play critical roles in marine nutrient cycles by incorporating and redistributing dissolved organic matter (DOM) and inorganic nutrients in the ocean. TonB-dependent transporter (TBDT) proteins allow Gram-negative bacteria to take up scarce resources from nutrient-limiting environments as well as siderophores, heme, vitamin B12, and recently identified carbohydrates. Thus, the characterization of TBDT distribution and functions is essential to better understand the contribution TBDT to DOM assimilation and its consequences on nutrient cycling in the environment.

Methodology/Principal Findings

This study presents the distribution of encoded known and putative TBDT proteins in the genomes of microorganisms and from the Global Ocean Survey data. Using a Lek clustering algorithm and substrate specificities, the TBDT sequences were mainly classified into the following three groups: (1) DOM transporters; (2) Siderophores/Vitamins transporters; and (3) Heme/Hemophores/Iron(heme)-binding protein transporters. Diverse TBDTs were found in the genomes of oligotroph Citromicrobium bathyomarinum JL354 and Citromicrobium sp JLT1363 and were highly expressed in the stationary phase of bacterial growth. The results show that the Gammaproteobacteria and the Cytophaga-Flavobacterium-Bacteroides (CFB) group bacteria accounted for the majority of the TBDT gene pool in marine surface waters.

Conclusions/Significance

The results of this study confirm the ecological importance of TBDTs in DOM assimilation for bacteria in marine environments owing to a wide range of substrate utilization potential in the ubiquitous Gammaproteobacteria and CFB group bacteria.