Extracellular hydrolytic enzyme screening of culturable heterotrophic bacteria from deep-sea sediments of the Southern Okinawa Trough

The Southern Okinawa Trough is an area of focused sedimentation due to particulate matter export from the shelf of the East China Sea and the island of Taiwan. In order to understand the geomicrobiological characteristics of this unique sedimentary environment, bacterial cultivations were carried out for an 8.61 m CASQ core sediment sample. A total of 98 heterotrophic bacterial isolates were characterized based on 16S rRNA gene phylogenetic analysis. These isolates can be grouped into four bacterial divisions, including 13 genera and more than 20 species. Bacteria of the gamma-Proteobacteria lineage, especially those from the Halomonas ( 27 isolates) and Psychrobacter ( 20 isolates) groups, dominate in the culturable bacteria assemblage. They also have the broadest distribution along the depth of the sediment. More than 72.4% of the isolates showed extracellular hydrolytic enzyme activities, such as amylases, proteases, lipases and Dnases, and nearly 59.2% were cold-adapted exoenzyme-producers. Several Halomonas strains show almost all the tested hydrolases activities. The wide distribution of exoenzyme activities in the isolates may indicate their important ecological role of element biogeochemical cycling in the studied deep-sea sedimentary environment.

Black carbon and polycyclic aromatic hydrocarbons (PAHs) in surface sediments of China's marginal seas

This study investigates the distribution of black carbon (BC) and its correlation with total polycyclic aromatic hydrocarbons (I PAH) pound in the surface sediments of China's marginal seas. BC content ranges from < 0.10 to 2.45 mg/g dw (grams dry weight) in the sediments studied, and varied among the different coastal regions. The Bohai Bay sediments had the highest BC contents (average 2.18 mg/g dw), which comprises a significant fraction (27%-41%) of the total organic carbon (TOC) preserved in the sediments. In comparison, BC in the surface sediments of the North Yellow Sea, Jiaozhou Bay, East China Sea and the South China Sea is less abundant and accounted for an average of 6%, 8%, 14% and 5%, respectively, of the sedimentary organic carbon pool. The concentration of I PAH pound in the surface sediments ranges from 41 to 3 667 ng/g dw and showed large spatial variations among the sampling sites of different costal regions. The Bohai Bay has the highest I PAH pound values, ranging from 79 to 3 667 ng/g dw. This reflects the high anthropogenically contaminated nature of the sediments in the bay. BC is positively correlated to TOC but a strong correlation is not found between BC and I PAH pound in the surface sediments studied, suggesting that BC and PAHs preserved in the sediments are derived from different sources and controlled by different biogeochemical processes. Our study suggests that the abundance of BC preserved in the sediments could represent a significant sink pool of carbon cycling in China's marginal seas.

Preservation of black carbon in the shelf sediments of the East China Sea

Concentrations and carbon isotopic (C-14, C-13) compositions of black carbon (BC) were measured for three sediment cores collected from the Changjiang River estuary and the shelf of the East China Sea. BC concentrations ranged from 0.02 to 0.14 mg/g (dry weight), and accounted for 5% to 26% of the sedimentary total organic carbon (TOC) pool. Among the three sediment cores collected at each site, sediment from the Changjiang River estuary had relatively high BC contents compared with the sediments from the East China Sea shelf, suggesting that the Changjiang River discharge played an important role in the delivery of BC to the coastal region. Radiocarbon measurements indicate that the ages of BC are in the range of 6910 to 12250 years old B. P. (before present), that is in general, 3700 to 9000 years older than the C-14 ages of TOC in the sediments. These variable radiocarbon ages suggest that the BC preserved in the sediments was derived from the products of both biomass fire and fossil fuel combustion, as well as from ancient rock weathering. Based on an isotopic mass balance model, we calculated that fossil fuel combustion contributed most (60%. 80%) of the BC preserved in these sediments and varied with depth and locations. The deposition and burial of this "slow-cycling" BC in the sediments of the East China Sea shelf represent a significant pool of carbon sink and could greatly influence carbon cycling in the region.

Forms and functions of inorganic carbon in the Jiaozhou Bay sediments

Inorganic carbon forms and their influencing factors, mutual transformation and contribution to carbon cycling in the Jiaozhou Bay sediments were discussed. The results show that inorganic carbon in sediments could be divided into five forms: NaCl form, NH3 center dot H2O form, NaOH form, NH2OH center dot HCl form and HCl form. Thereinto, NH2OH center dot HCl form and HCl form account for more than 70% of total inorganic carbon. There was close relationship among every form of inorganic carbon and their correlativity was clearly different with different sedimentary environment except the similar strong positive correlation among NH2OH center dot HCl form, HCl form and total inorganic carbon in all regions of the Jiaozhou Bay. All forms of inorganic carbon were influenced by organic carbon, pH, Eh, Es, nitrogen and phosphorus in sediments, but their influence had different characteristics in different regions. Every farm of inorganic carbon transformed into each other continuously during early diagenesis of sediments and the common phenomenon was that NaCl form, NH3 center dot H2O form, NaOH form and NH2OH center dot HCl form might transform into steady HCl form. NaCl form, NH3 center dot H2O form, NaOH form and NH2OH center dot HCl form could participate in carbon recycle and they are potential carbon source; HCl form may be buried for a long time in sediments, and it may be one of the final resting places of atmospheric CO2. Inorganic carbon which entered into sediments was about 4.98 x 10(10) g in the Jiaozhou Bay every year, in which about 1.47x10(10) g of inorganic carbon might be buried for a long time and about 3.51. x 10(10) g of inorganic carbon might return into seawater and take part in carbon recycling.

The nitrogen isotopic composition of dissolved nitrate in the Yangtze River (Changjiang) estuary, China

The nitrogen isotopic composition of dissolved nitrate (delta N-15-NO3-) in surface water of the Yangtze River estuary was determined in four seasons of 2006. delta N-15-NO3- ranged from 0.4 parts per thousand to 6.5 parts per thousand and varied with seasons and geographic regions, reflecting the dynamics of nitrogen cycling in the estuarine ecosystem. delta N-15-NO3- was markedly lower in February than in other seasons and exhibited conservative mixing, which was probably attributed to the NO3- being sourced from the atmospheric deposition and agricultural fertilizer. In the upper estuary, the influence of riverine inputs was important during all surveys. in the turbidity maximum zone, nitrification was found with nitrate depleted in N-15 in May, whereas denitrification resulting in heavy delta N-15-NO3- played an important role in August. More enriched delta N-15-NO3- values coinciding with losses of nitrate concentrations based on the conservative mixing model were found in the adjacent marine area in May, and may reflect obvious phytoplankton assimilation of dissolved nitrate. In this manner, delta N-15-NO3- may be a sensitive indicator of nitrogen sources and biogeochemical processing existing in this estuary in conjunction with the variations of dissolved nitrate and other environmental factors. (C) 2009 Elsevier Ltd. All rights reserved.

Geochemical characteristics of nitrogen in the southern Yellow Sea surface sediments

Roles and distributions of various forms of nitrogen in biogeochemical cycling in the southern Yellow Sea surface sediments were investigated. The southern Yellow Sea could be divided into three regions (I, II and III) according to the proportion of fine-grained sediment in > 65%, 35-65% and < 35%, respectively. The ratios of different forms of nitrogen contents between each two of the three regions indicated that the nitrogen contents increased with the proportion of fine-grained sediment increasing. The quanta of exchangeable forms of nitrogen were similar in the three regions, while their releasing time increased from regions I to III, indicating that the cycle of nitrogen in fine-grained sediments was shorter than that in coarse-grained sediments. Nitrogen burial fluxes were also similar in these regions, while the burial efficiency increased from regions I to III. The highest burial efficiency was 30.21% in region III, indicating that more than 70% of nitrogen in the southern Yellow Sea surface sediments could be released to take part in biogeochemical recycling. When all the four forms of exchangeable nitrogen (nitrogen in ion exchangeable form (IEF-N), nitrogen in weak acid extractable form (WAEF-N), nitrogen in strong alkali extractable form (SAEF-N) and nitrogen in strong oxidant extractable form (SOEF-N)) were released to take part in recycling, their potential contributions were 80% (SOEF-N), 11% (IEF-N), 6% (SAEF-N), 3% (WAEF-N) respectively, which showed that SOEF-N was the predominant one, and its contribution to biogeochemical cycling was the highest. (c) 2004 Elsevier B.V. All rights reserved.

Determination for the different combined-form inorganic carbon in marine sediments

Marine sediment is the important sources and sinks of carbon. The inorganic carbon(IC) in marine sediments plays an important role in carbon cycling. In order to understand IC function in carbon cycling, sequential extraction method based on IC combined chemical strength difference were established to get five phases: NaCl phase (step I), NH3 - H2O phase (step II), NaOH phase (step III), NH2OH . HCl phase (step IV) and HCl phase(step V). The best extraction conditions were obtained by a series of experiments. Extractants were added into plastic centrifuge tubes in Step I - M, the capped tube were placed on a shaker table to keep the solids suspended for two hours. The suspended solution was separated by centrifugal, the residues were washed with water. The two supernatant were combined and the CO, was finally determined by volumetric analysis. The residues were transferred into conical flask in step IV and V, and then the extractants were added. The produced CO2 was adsorbed by saturated Ba(OH)(2) solution, and determined by volumetric analysis. This method for IC has a good precision in the analysis sediment samples.

A simple and accurate method for determining accurately dissolved inorganic carbon in seawaters

Dissolved inorganic carbon (DIC) account for more than 95% of total carbon in seawater, so it is necessary to make reliable and precise measurements of DIC to study marine carbon cycling. In order to establish a simple and speed method, an airproof device of gas extraction-absorption was designed. Finally a simple method was developed for the determination of DIC in seawater through a large mount of experiments. The determination procedure is as follows: 100 similar to 150 mL seawater was put into conical flask, then add 10% H3PO4, the DIC in seawater sample was dissolved to form CO2 gas and carried by pure N-2, then the CO2 gas was absorbed by two grades 0.1 mol/L NaOH solution. Finally the absorbed solution was titrated by HCl standard solution of 0.01000 mol/L with the end points detected with the indicator phenolphthalein and bromocresol green-methyl red mixture. The precision and accuracy of the method were satisfied. This method was used to analyse seawater samples from Jiaozhou bay in June, 2003. The result shows that the average DIC in surface seawater is 2066 mumol/L, DIC in bottom seawater is 2075 mumol/L inside bay, but the average DIC in surface seawater is 1949 mumol/L, DIC in bottom seawater is 2147 mumol/L outside bay.

Sources and transport of dissolved and particulate organic carbon in the Mississippi River estuary and adjacent coastal waters of the northern Gulf of Mexico

Dissolved organic carbon (DOC), stable carbon isotopic (delta(13)C) compositions of DOC and particulate organic carbon (POC), and elemental C/N ratios of POC were measured for samples collected from the lower Mississippi and Atchafalaya rivers and adjacent coastal waters in the northern Gulf of Mexico during the low flow season in June 2000 and high flow season in April 2001. These isotopic and C/N results combined with DOC measurements were used to assess the sources and transport of terrestrial organic matter from the Mississippi and Atchafalaya rivers to the coastal region in the northern Gulf of Mexico. delta(13)C values of both POC (-23.8parts per thousand to -26.8parts per thousand) and DOC (-25.0parts per thousand to -29.0parts per thousand) carried by the two rivers were more depleted than the values measured for the samples collected in the offshore waters. Strong seasonal variations in delta(13)C distributions were observed for both POC and DOC in the surface waters of the region. Fresh water discharge and horizontal mixing played important roles in the distribution and transport of terrestrial POC and DOC offshore. Our results indicate that both POC and DOC exhibited non-conservative behavior during the mixing especially in the mid-salinity range. Based on a simple two end-member mixing model, the comparison of the measured DOC-delta(13)C with the calculated conservative isotopic mixing curve indicated that there was a significant in situ production of marine-derived DOC in the mid- to high-salinity waters consistent with our in situ chlorophyll-a measurements. Our DOC-delta(13)C data suggest that a removal of terrestrial DOC mainly occurred in the high-salinity (>25) waters during the mixing. Our study indicates that the mid- to high- (10-30) salinity range was the most dynamic zone for organic carbon transport and cycling in the Mississippi River estuary. Variability in isotopic and elemental compositions along with variability in DOC and POC concentrations suggest that autochthonous production, bacterial utilization, and photo-oxidation could all play important roles in regulating and removing terrestrial DOC in the northern Gulf of Mexico and further study of these individual processes is warranted. (C) 2004 Elsevier B.V. All rights reserved.

Bacterial roles in the formation of high-molecular-weight dissolved organic matter in estuarine and coastal waters: Evidence from lipids and the compound-specific isotopic ratios

High-molecular-weight dissolved organic matter (HMW-DOM, > 1,000 Daltons) is actively involved in the global biogeochemical cycling of many elements, but its carbon sources and detailed formation pathways are still not well understood. In this study, we measured bulk stable carbon and nitrogen isotopic ratios, lipid composition, and compound-specific carbon isotopic ratios of HMW-DOM samples collected from four U.S. estuaries (Boston Harbor/Massachusetts Bay, Delaware/Chesapeake Bay, San Diego Bay, and San Francisco Bay). Analytical results show (1) a fraction of HMW-DOM (lipid associated) in estuarine and coastal waters is derived from bacteria and phytoplankton; (2) this fraction of HMW-DOM is formed by various release processes of bacterial membrane components and bacterial reworking of phytoplankton-derived material; (3) this fraction of HMW-DOM is generally present in all samples from different coastal systems despite variable organic matter inputs and environmental conditions, suggesting an important bacterial role in HMW-DOM formation.

Sources and preservation of organic matter in Plum Island salt marsh sediments (MA, USA): long-chain n-alkanes and stable carbon isotope compositions

Elemental (TOC, TN, C/N) and stable carbon isotopic (delta(13)C) compositions and n-alkane (nC(16-38)) concentrations were measured for Spartina alterniflora, a C-4 marsh grass, Typha latifolia, a C-3 marsh grass, and three sediment cores collected from middle and upper estuarine sites from the Plum Island salt marshes. Our results indicated that the organic matter preserved in the sediments was highly affected by the marsh plants that dominated the sampling sites. delta(13)C values of organic matter preserved in the upper fresh water site sediment were more negative (-23.0+/-0.3) as affected by the C-3 plants than the values of organic matter preserved in the sediments of middle (-18.9+/-0.8) and mud flat sites (-19.4+/-0.1) as influenced mainly by the C4 marsh plants. The distribution of n-alkanes measured in all sediments showed similar patterns as those determined in the marsh grasses S. alterniflora and T. latifolia, and nC(21) to nC(33) long-chain n-alkanes were the major compounds determined in all sediment samples. The strong odd-to-even carbon numbered n-alkane predominance was found in all three sediments and nC(29) was the most abundant homologue in all samples measured. Both delta(13)C compositions of organic matter and n-alkane distributions in these sediments indicate that the marsh plants could contribute significant amount of organic matter preserved in Plum Island salt marsh sediments. This suggests that salt marshes play an important role in the cycling of nutrients and organic carbon in the estuary and adjacent coastal waters. (C) 2003 Elsevier Ltd. All rights reserved.

Characteristics of nitrogen forms in the surface sediments of southwestern Nansha Trough, South China Sea

The area of the southwestern Nansha Trough is one of the most productive areas of the southern South China Sea. It is a typical semi-deep sea area of transition from shoal to abyssal zone. To understand distributions and roles of nitrogen forms involved in biogeochemical cycling in this area, contents of nitrogen in four extractable forms: nitrogen in ion exchangeable form (IEF-N), nitrogen in weak acid extractable form (WAEF-N), nitrogen in strong alkali extractable form (SAEF-N) and nitrogen in strong oxidation extractable form (SOEF-N), as well as in total nitrogen content (TN) in surface sediments were determined from samples collected from the cruise in April-May 1999. The study area was divided into three regions (A, B and C) in terms of clay sediment (< 4 mu m) content at < 40%, 40%-60% and > 60%, respectively. Generally, region C was the richest in the nitrogen of all forms and region A the poorest, indicating that the finer the grain size is, the richer the contents of various nitrogen are. The burial efficiency of total nitrogen in surface sediments was 28.79%, indicating that more than 70% of nitrogen had been released and participated in biogeochemical recycling through sediment-water interface.

Inorganic carbon of sediments in the Yangtze River Estuary and Jiaozhou Bay

JGOFS results showed that the ocean is a major sink for the increasing atmospheric carbon dioxide resulting from human activity. However, the role of the coastal seas in the global carbon cycling is poorly understood. In the present work, the inorganic carbon (IC) in the Yangtze River Estuary and Jiaozhou Bay are studied as examples of offshore sediments. Sequential extraction was used to divide inorganic carbon in the sediments into five forms, NaCl form, NH3 H2O form, NaOH form, NH2OH HCl form and HCl form. Studied of their content and influencing factors were also showed that NaCl form < NH3 H2O form < NaOH form < NH2OH HCl form < HCl form, and that their influencing factors of pH, Eh, Es, water content, organic carbon, organic nitrogen, inorganic nitrogen, organic phosphorus and inorganic phosphorus on inorganic carbon can be divided into two groups, and that every factor has different influence on different form or on the same form in different environment. Different IC form may transform into each other in the early diagenetic process of sediment, but NaCl form, NH3 H2O form, NaOH form and NH2OH HCl form may convert to HCl form ultimately. So every IC form has different contribution to carbon cycling. This study showed that the contribution of various form of IC to the carbon cycle is in the order of NaOH form > NH2OH HCl form > NH3 H2O form > NaCl form > HCl form, and that the contribution of HCl form contributes little to carbon cycling, HCl form may be one of end-result of atmospheric CO2. So Yangtze River estuary sediment may absorb at least about 40.96x10(11) g atmospheric CO2 every year, which indicated that offshore sediment play an important role in absorbing atmospheric CO2.

The behaviors and sources of dissolved arsenic and antimony in Bohai Bay

In this paper, we determined the concentrations of antimony species (antimonite (Sb(III)), antimonate (Sb(V)) and dissolved inorganic antimony (DISb)) and arsenic, in Bohai Bay seawaters, as well as the relationships of the analytes with environmental factors such as seawater characteristics (e.g., suspended particulate material (SPM), salinity and total organic carbon (TOC)), heavy metals, nutrients and phytoplankton species, and evaluated the sources of arsenic and antimony. Dissolved arsenic and antimony concentrations in the surface waters were ranging spatially from 1.03 to 1.26 ng/ml and 0.386 to 1.075 ng/ml, with mean values of 1.18 and 0.562 ng/ml, respectively. Sb(V) as the prominent chemical species constituted about 89%. Regarding arsenic concentrations in the surface waters, there was a tendency for a small variation. However, antimony species concentrations were much variable than arsenic. The highest arsenic and antimony concentrations were found near the Haihe Estuary. These distribution patterns were controlled mainly by environmental factors, biological activities and sources. In this region, DISb and Sb(V) negatively correlated with salinity. Besides, arsenic and antimony correlated well with the nutrients, chlorophyll a and phytoplankton, implying that arsenic and antimony had been involved in biological cycling. In addition, according to our estimate, about 333.5 x 10(8) mg/year of arsenic and 454.2 x 10(8) mg/year of antimony reached Bohai Bay via rivers. (C) 2010 Elsevier Ltd. All rights reserved.

Variability in radiocarbon ages of biochemical compound classes of high molecular weight dissolved organic matter in estuaries

High molecular weight dissolved organic matter (HMW-DOM, > 1000 Da) represents a major fraction (> 30%) of dissolved organic carbon (DOC) in the ocean and thus plays an important role in the global biogeochemical cycling of carbon and many other elements. Its organic sources and formation mechanisms, however, are still not well understood especially in estuarine and coastal regions where multiple natural and anthropogenic sources contribute to total HMW-DOM. In this paper we report our measurements of natural radiocarbon (C-14) abundances and stable carbon isotope (C-13) compositions of the major biochemical compound classes: amino acids, carbohydrates and lipids separated from eight HMW-DOM samples collected from five US estuaries as part of our on-going study of sources, distribution and transport of chromophoric dissolved organic matter (CDOM) in estuarine and coastal waters. Distinct differences in both C-14 and C-13 values were found among the bulk HMW-DOM samples as well as the individual compound classes. Radiocarbon ages of the major compound classes varied by as much as 27,000 years in a single sample. The calculated average radiocarbon ages of the compound fractions of HMW-DOM indicate that the total lipid fraction is very "old", while the acid-insoluble fraction is slightly younger. Total amino acid and carbohydrate fractions, however, have relatively modern apparent C-14 ages. The significant variability in C-14 ages among the compound classes indicates not only multiple organic carbon sources but also different formation and turnover pathways controlling the cycling of different biochemical components of HMW-DOM in estuarine and coastal waters. (c) 2006 Elsevier Ltd. All rights reserved.