Speleothems from Sahastradhara Caves in Siwalik Himalaya,India: Possible Biogenic Inputs

Stalactites and moonmilk from Sahastradhara caves in Siwalik Himalayas were studied to understand the role of microbes in their genesis. Fourier spectroscopy in the moonmilk indicates a complex milieu of organic compounds that is unusual for inorganic formations. Stable C and O isotopes show trends in the moonmilk and stalactite, which suggest biogenic input; the geochemical inference is consistent with evidence from microscopy and laboratory-based microbial cultures. Light microscopy of moonmilk samples show the presence of a number of microbial forms similar to Cyanobacteria, and scanning electron microscope (SEM) images show microbial structures similar to Spirulina. The total number of microbial cells using SYBR Gold is 6.5 × 10⁵ cells, g sed^?1in moonmilk and 3.2 × 10⁵ cells, g sed^?1 in stalactites. FISH indicates approximately 3.5 × 10⁵ cells, g sed^?1 in moonmilk and 2 × 10⁵ cells, g sed^?1 in stalactites. SEM images of the moonmilk indicate a large network of microbial filaments along with minerals, which are identified as calcite based on their x-ray diffraction pattern. In vitro laboratory cultures with pure monogenic strains isolated from the moonmilk and stalactites raise pH in the medium, which facilitate calcite precipitation. The mineral precipitating isolates were identified as: Bacillus pumilis, B. cereus, B. anthracis, B. lentus, B. sphaericus, B. circulans and Actinomycetes. The Sahastradhara moonmilk and statactites are colonized by a diverse microbial community and the isolated bacterial strains induce biomineralization on different nutrient media, supporting their biogenic origin.     

Evaluation of commercial and natural food in experimental cultures of white shrimp based on stable carbon isotopes

The farming of shrimp is developing quickly worldwide, and recently, ingredients such as seaweeds in low proportion (25 to 4 %), incorporated into the commercial food, have been shown to improve the shrimp productive variables. The change of commercial foods to commercial feed with a proportion of natural food, and finally, to natural food has been little and simultaneously evaluated. The aim of our study was to determine the relative contribution of dietary carbon to the growth of Litopenaeus vannamei fed with a proportion of 4 % Sargassum (δ¹³C = ?20.9?±?0.05?‰), 4 % Ulva (δ¹³C = ?20.6?±?0.6?‰) meal, and a control diet (δ¹³C?=??22.6?±?0.2?‰) in 60-L tanks for 30 days, and finally, with the green seaweed Ulva spp. (δ¹³C = ?13.2?±?0.25?‰) and Ulva meal (δ¹³C = ?14.5?±?0.6?‰) in open-air ponds for 120 days, by measuring δ¹³C for each of the foods and in the muscle of shrimp. After 15 days, the rates of metabolic turnover (Δ¹³C = δ¹³C_shrimp ? δ¹³C_food) were constant until the end of the experiment in the tanks. The muscle of shrimp assimilated carbon from all diets, which demonstrated the potential use of combined diets and the optimization of their use in diets containing seaweed. Our data will be useful in future interpretations of field and laboratory isotopic values for this species.     

Bacterial abundance and composition in marine sediments beneath the Ross Ice Shelf,Antarctica

Marine sediments of the Ross Sea, Antarctica, harbor microbial communities that play a significant role in the decomposition, mineralization, and recycling of organic carbon (OC). In this study, the cell densities within a 153‐cm sediment core from the Ross Sea were estimated based on microbial phospholipid fatty acid (PLFA) concentrations and acridine orange direct cell counts. The resulting densities were as high as 1.7 × 10⁷ cells mL^?1 in the top ten centimeters of sediments. These densities are lower than those calculated for most near‐shore sites but consistent with deep‐sea locations with comparable sedimentation rates. The δ¹³C measurements of PLFAs and sedimentary and dissolved carbon sources, in combination with ribosomal RNA (SSU rRNA) gene pyrosequencing, were used to infer microbial metabolic pathways. The δ¹³C values of dissolved inorganic carbon (DIC) in porewaters ranged downcore from ?2.5‰ to ?3.7‰, while δ¹³C values for the corresponding sedimentary particulate OC (POC) varied from ?26.2‰ to ?23.1‰. The δ¹³C values of PLFAs ranged between ?29‰ and ?35‰ throughout the sediment core, consistent with a microbial community dominated by heterotrophs. The SSU rRNA gene pyrosequencing revealed that members of this microbial community were dominated by β‐, δ‐, and γ‐Proteobacteria, Actinobacteria, Chloroflexi and Bacteroidetes. Among the sequenced organisms, many appear to be related to known heterotrophs that utilize OC sources such as amino acids, oligosaccharides, and lactose, consistent with our interpretation from δ¹³C_PLFA analysis. Integrating phospholipids analyses with porewater chemistry, δ¹³C_DIC and δ¹³C_POC values and SSU rRNA gene sequences provides a more comprehensive understanding of microbial communities and carbon cycling in marine sediments, including those of this unique ice shelf environment.     

Kangaroo tooth enamel oxygen and carbon isotope variation on a latitudinal transect in southern Australia: implications for palaeoenvironmental reconstruction

Tooth enamel apatite carbonate carbon and oxygen isotope ratios of modern kangaroos (Macropus spp.) collected on a 900-km latitudinal transect spanning a C₃–C₄ transition zone were analysed to create a reference set for palaeoenvironmental reconstruction in southern Australia. The carbon isotope composition of enamel carbonate reflects the proportional intake of C₃ and C₄ vegetation, and its oxygen isotope composition reflects that of ingested water. Tooth enamel forms incrementally, recording dietary and environmental changes during mineralisation. Analyses show only weak correlations between climate records and latitudinal changes in δ¹³C and δ¹⁸O. No species achieved the δ¹³C values (~?1.0 ‰) expected for 100 % C₄ grazing diets; kangaroos at low latitudes that are classified as feeding primarily on C₄ grasses (grazers) have δ¹³C of up to ?3.5 ‰. In these areas, δ¹³C below ?12 ‰ suggests a 100 % C₃ grass and/or leafy plant (browse) diet while animals from higher latitude have lower δ¹³C. Animals from semi-arid areas have δ¹⁸O of 34–40 ‰, while grazers from temperate areas have lower values (~28–30 ‰). Three patterns with implications for palaeoenvironmental reconstruction emerge: (1) all species in semi-arid areas regularly browse to supplement limited grass resources; (2) all species within an environmental zone have similar carbon and oxygen isotope compositions, meaning data from different kangaroo species can be pooled for palaeoenvironmental investigations; (3) relatively small regional environmental differences can be distinguished when δ¹³C and δ¹⁸O data are used together. These data demonstrate that diet–isotope and climate–isotope relationships should be evaluated in modern ecosystems before application to the regional fossil record.     

Thermophilic Archaeal Diversity and Methanogenesis from El Tatio Geyser Field,Chile

The hydrothermal fluid chemistry at El Tatio Geyser Field (ETGF) in northern Chile suggests that biogenic CO₂–CH₄ cycling may play an important role in water chemistry, and relatively low sulfate (0.6–1 mM) and high molecular hydrogen (H₂) concentrations (67–363 nM) suggest that methanogenic Archaea are present in ETGF microbial mats. In this study, δ¹³C analysis of dissolved inorganic carbon and methane was not indicative of biogenic methane production (δ¹³C_CH4 values ranging from ?15‰ to ?5.3‰); however, methanogenic Archaea were successfully cultured from each of the hydrothermal sites sampled. Sanger sequencing using universal Archaea primers identified putative methanogenic orders with varying metabolic capabilities, including Methanobacteriales, Methanomicrobiales and Methanosarcinales.     

Three years of soil respiration in a mature eucalypt woodland exposed to atmospheric CO<Subscript>2</Subscript> enrichment

Large amounts of atmospheric N deposition cause negative effects on ecosystems. Effective mitigation strategies require the sources of N deposition to be identified and the contributions from individual sources to be quantified. Determination of the isotopic composition represents a useful approach in source apportionment. In this study, the δ¹⁵N-NH_x of wet and dry atmospheric deposition and the main NH₃ emission sources were analyzed at an urban, a suburban and a rural site in the Taihu Lake region of China. The 2-year average δ¹⁵N-\( {\text{NH}}_{4}^{ + } \) of precipitation was ? 3.0 ± 2.3, ? 3.1 ± 2.8 and ? 0.5 ± 2.8‰ for the urban, suburban and rural sites, respectively. These values were much lower than the corresponding values for particulate \( {\text{NH}}_{4}^{ + } \) (15.9, 15.2 and 14.3‰ at the urban, suburban and rural sites, respectively), and much higher than those of gaseous δ¹⁵N-NH₃ (? 16.7, ? 18.2 and ? 17.4‰ at the urban, suburban and rural sites, respectively). The δ¹⁵N-NH₃ of NH₃ from the main emission sources ranged from ? 30.8 to ? 3.3‰ for volatilized fertilizer, from ? 35.1 to ? 10.5‰ for emissions from a pig farm, and ? 24.7 to ? 11.3‰ for emissions from a dairy farm. Temporal variations of deposition δ¹⁵N-NH_x indicated that δ¹⁵N-NH_x values were lower in summer and autumn, but higher in winter and spring for both precipitation \( {\text{NH}}_{4}^{ + } \)-N and gaseous NH₃-N. Weather conditions such as temperature and precipitation significantly influenced the spatial and temporal distribution of isotope values of the deposition. Analysis of δ¹⁵N-NH_x in deposition and emission sources identified volatilized fertilizer and livestock wastes as the origins of both gaseous NH₃-N and precipitation \( {\text{NH}}_{4}^{ + } \)-N over the region. A stable isotope mixing model estimated that volatilized fertilizer and animal excreta contributed more than 65% to precipitation \( {\text{NH}}_{4}^{ + } \)-N, more than 60% to particulate \( {\text{NH}}_{4}^{ + } \)-N, and more than 75% to gaseous NH₃-N.     

Induction of metamorphosis of pediveliger larvae of the mussel Mytilus galloprovincialis Lamarck, 1819 using neuroactive compounds,KCl, NH4Cl and organic solvents

Pediveliger larvae of Mytilus galloprovincialis were subjected to a series of bioassays to investigate the induction of metamorphosis using neuroactive compounds, K⁺, NH₄ ⁺ and organic solvents. Growth and survival of post-larvae obtained using ethanol and methanol were also observed. Epinephrine, phenylephrine, clonidine and metanephrine induced larval metamorphosis at 10^?6 to 10^?4 M in both 24-h and continuous exposure assays. In 24-h exposure assays, α-methyldopa at 5×10^?5 M and methoxyphenamine at 5×10^?5?10^?4 M induced 55?94% metamorphosis. Similarly, excess K⁺ at 3×10^?2 M induced 39% metamorphosis and NH₄ ⁺ at 1?5×10^?2 M induced 63–78% metamorphosis. The EC50s of seven organic solvents ranged from 0.04 to 0.82 M. Post-larvae that metamorphosed using ethanol and methanol survived as juveniles and grew at the same rate as those from microbial biofilm. Thus, the above compounds can be useful inducers of metamorphosis for antifouling studies using larvae and juveniles of M. galloprovincialis.     

Effects of land use on sources and ages of inorganic and organic carbon in temperate headwater streams

The amounts, sources and relative ages of inorganic and organic carbon pools were assessed in eight headwater streams draining watersheds dominated by either forest, pasture, cropland or urban development in the lower Chesapeake Bay region (Virginia, USA). Streams were sampled at baseflow conditions six different times over 1 year. The sources and ages of the carbon pools were characterized by isotopic (δ¹³C and ?¹⁴C) analyses and excitation emission matrix fluorescence with parallel factor analysis (EEM–PARAFAC). The findings from this study showed that human land use may alter aquatic carbon cycling in three primary ways. First, human land use affects the sources and ages of DIC by controlling different rates of weathering and erosion. Relative to dissolved inorganic carbon (DIC) in forested streams which originated primarily from respiration of young, ¹⁴C-enriched organic matter (OM; δ¹³C = ?22.2 ± 3 ‰; ?¹⁴C = 69 ± 14 ‰), DIC in urbanized streams was influenced more by sedimentary carbonate weathering (δ¹³C = ?12.4 ± 1 ‰; ?¹⁴C = ?270 ± 37 ‰) and one of pasture streams showed a greater influence from young soil carbonates (δ¹³C = ?5.7 ± 2.5 ‰; ?¹⁴C = 69 ‰). Second, human land use alters the proportions of terrestrial versus autochthonous/microbial sources of stream water OM. Fluorescence properties of dissolved OM (DOM) and the C:N of particulate OM (POM) suggested that streams draining human-altered watersheds contained greater relative contributions of DOM and POM from autochthonous/microbial sources than forested streams. Third, human land uses can mobilize geologically aged inorganic carbon and enable its participation in contemporary carbon cycling. Aged DOM (?¹⁴C = ?248 to ?202 ‰, equivalent¹⁴C ages of 1,811–2,284 years BP) and POM (?¹⁴C = ?90 to ?88 ‰, ¹⁴C ages of 669–887 years BP) were observed exclusively in urbanized streams, presumably a result of autotrophic fixation of aged DIC (?297 to ?244 ‰, ¹⁴C age = 2,251–2,833 years BP) from sedimentary shell dissolution and perhaps also watershed export of fossil fuel carbon. This study demonstrates that human land use may have significant impacts on the amounts, sources, ages and cycling of carbon in headwater streams and their associated watersheds.     

Facies development and sequence stratigraphy of the Ludfordian (Upper Silurian) deposits in the Zbruch River Valley, Podolia, western Ukraine: local facies overprint on the δ13Ccarb record of a global stable carbon isotope excursion

The Ludfordian (Upper Silurian) succession in Podolia, western Ukraine, represents a Silurian carbonate platform developed in an epicontinental sea on the shelf of the paleocontinent of Baltica. Coeval deposits throughout this basin record a positive stable carbon isotope excursion known as the Lau excursion. The record of this excursion in Podolia exhibits an unusual amplitude from highly positive (+6.9 ‰) to highly negative (?5.0 ‰) δ¹³C_carb values. In order to link δ¹³C_carb development with facies, five sections in the Zbruch River Valley were examined, providing microfacies characterization and revised definitions of the Isakivtsy, Prygorodok, and Varnytsya Formations. The Isakivtsy Fm. is developed as dolosparite replacing originally bioclastic limestone. The Prygorodok Fm., recording strongly depleted (down to ?10.53 ‰) to near zero (0.12 ‰) δ¹³C_carb values is developed as laminated, organic-rich dolomicrite with metabentonite and quartz siltstone beds. The Varnytsya Fm. is characterized by peritidal deposition with consistent, slightly negative δ¹³C_carb values (?0.57 to ?3.20 ‰). It is proposed that dolomitization of the Isakivtsy Fm. is associated with a sequence boundary and erosional surface. The overlying Prygorodok Fm. represents the proximal part of a TST deposited in restricted and laterally extremely variable environments dominated by microbial carbonate production. The transition to the overlying Varnytsya Fm. facies is marked by a maximum flooding surface. The SB and MFS are potentially correlative within the basin and support a global rapid sea-level fall previously proposed for this interval. The interpretation of the Prygorodok Fm. as coastal lake deposits may explain the unusual δ¹³C_carb values and constitute one of the few records of this type of environment identified in the early Paleozoic.     

Fluxes of nitrous oxide, methane and carbon dioxide during freezing–thawing cycles in an Inner Mongolian steppe

Combined measurements of nitrification activity and N₂O emissions were performed in a lowland and a montane tropical rainforest ecosystem in NE-Australia over a 18 months period from October 2001 until May 2003. At both sites gross nitrification rates, measured by the BaPS technique, showed a strong seasonal pattern with significantly higher rates of gross nitrification during wet season conditions. Nitrification rates at the montane site (1.48?±?0.24–18.75?±?2.38 mg N kg^?1 day^?1) were found to be significantly higher than at the lowland site (1.65?±?0.21–4.54?±?0.27 mg N kg^?1 day^?1). The relationship between soil moisture and gross nitrification rates could be described best by O’Neill functions having a soil moisture optimum of nitrification at app. 65% WFPS. At the lowland site, for which continuous measurements of N₂O emissions were available, nitrification was positively correlated with N₂O emission. Nitrification contributed significantly to N₂O formation during dry season (app.85%) but less (app. 30%) during wet season conditions. In average 0.19‰ of the N metabolized by nitrification was released as N₂O. The N₂O fraction loss for nitrification was positively correlated with changes in soil moisture and varied slightly between 0.15 and 0.22‰. Our results demonstrate that combined N₂O emission and microbial N turnover studies covering prolonged observation periods are needed to clarify and quantify the role of the microbial processes nitrification and denitrification for annual N₂O emissions from soils of terrestrial ecosystems.     

Biomineralization Abilities of Cupriavidus Strain and Bacillus subtilis Strains In Vitro Isolated from Speleothems,Rani Cave,Chhattisgarh, India

In vitro culture experiments using three bacterial strains CSJC1, CSJC2, and CSJC3 isolated from speleothems, Rani cave, Chhattisgarh, India, were studied to examine their biomineralization potential. These speleothems showed high microbial cell enumerations on nutrient agar and iron agar (9 × 10⁴ CFU/g) followed by thiosulfate agar (7 × 10⁴ CFU/g), and 60 diverse strains were isolated. The BLASTn sequence search of 16S rRNA sequences with the NCBI database to establish the identity of CSJC1, CSJC2, and CSJC3 strains yielded similarity scores of ≥99% with the respective organisms, and the strains were identified as CSJC1 – Bacillus sp., CSJC2 – Cupriavidus sp., CSJC3 – Bacillus sp. The phylogenetic analysis of CSJC2 strain suggests that it formed a separate major cluster with Cupriavidus sp. and Cupriavidus necator. The phylogenetic analysis of CSJC1 and CSJC3 strains revealed that it formed a major cluster with several strains of Bacillus sp. and Bacillus subtilis. The biominerals induced by Cupriavidus sp. CSJC2 strain imaged with an ultra high-resolution field emission scanning electron microscope (FE-SEM) were seen as calcified coccoid shells that transformed into calcified dumbbells. FE-SEM imaging of biominerals induced by B. subtilis CSJC1 and CSJC3 tested both on B4 media and sheep blood agar individually showed that the precipitates formed calcified dumbbells that were almost similar but not identical phenotypically, indicating that strain-specific morphologies and crystal formation is easier when Ca is present in the media. This is the first comprehensive report on the possible evidences about the role of Cupriavidus sp. in calcite precipitation isolated from speleothems in the Indian caves. These results allow us to postulate that the identified strains may have a role in the biogenic influences in mineral formations at Rani cave.     

A preliminary survey on the occurrence of mycotoxigenic fungi and mycotoxins contaminating red rice at consumer level in Selangor, Malaysia

Red rice is a fermented product of Monascus spp. It is widely consumed by Malaysian Chinese who believe in its pharmacological properties. The traditional method of red rice preparation disregards safety regulation and renders red rice susceptible to fungal infestation and mycotoxin contamination. A preliminary study was undertaken aiming to determine the occurrence of mycotoxigenic fungi and mycotoxins contamination on red rice at consumer level in Selangor, Malaysia. Fifty red rice samples were obtained and subjected to fungal isolation, enumeration, and identification. Citrinin, aflatoxin, and ochratoxin-A were quantitated by ELISA based on the presence of predominant causal fungi. Fungal loads of 1.4?×?10⁴ to 2.1?×?10⁶?CFU/g exceeded Malaysian limits. Monascus spp. as starter fungi were present in 50 samples (100 %), followed by Penicillium chrysogenum (62 %), Aspergillus niger (54 %), and Aspergillus flavus (44 %). Citrinin was present in 100 % samples (0.23–20.65 mg/kg), aflatoxin in 92 % samples (0.61–77.33 μg/kg) and Ochratoxin-A in 100 % samples (0.23–2.48 μg/kg); 100 % citrinin and 76.09 % aflatoxin exceeded Malaysian limits. The presence of mycotoxigenic fungi served as an indicator of mycotoxins contamination and might imply improper production, handling, transportation, and storage of red rice. Further confirmatory analysis (e.g., HPLC) is required to verify the mycotoxins level in red rice samples and to validate the safety status of red rice.     

Isotopic biosignatures in carbonate‐rich,cyanobacteria‐dominated microbial mats of the Cariboo Plateau,B.C.

Photosynthetic activity in carbonate‐rich benthic microbial mats located in saline, alkaline lakes on the Cariboo Plateau, B.C. resulted in pCO₂ below equilibrium and δ¹³C_DIC values up to +6.0‰ above predicted carbon dioxide (CO₂) equilibrium values, representing a biosignature of photosynthesis. Mat‐associated δ¹³C_carb values ranged from ~4 to 8‰ within any individual lake, with observations of both enrichments (up to 3.8‰) and depletions (up to 11.6‰) relative to the concurrent dissolved inorganic carbon (DIC). Seasonal and annual variations in δ¹³C values reflected the balance between photosynthetic ¹³C‐enrichment and heterotrophic inputs of ¹³C‐depleted DIC. Mat microelectrode profiles identified oxic zones where δ¹³C_carb was within 0.2‰ of surface DIC overlying anoxic zones associated with sulphate reduction where δ¹³C_carb was depleted by up to 5‰ relative to surface DIC reflecting inputs of ¹³C‐depleted DIC. δ¹³C values of sulphate reducing bacteria biomarker phospholipid fatty acids (PLFA) were depleted relative to the bulk organic matter by ~4‰, consistent with heterotrophic synthesis, while the majority of PLFA had larger offsets consistent with autotrophy. Mean δ¹³C_org values ranged from ?18.7 ± 0.1 to ?25.3 ± 1.0‰ with mean Δ¹³C_inorg‐org values ranging from 21.1 to 24.2‰, consistent with non‐CO₂‐limited photosynthesis, suggesting that Precambrian δ¹³C_org values of ~?26‰ do not necessitate higher atmospheric CO₂ concentrations. Rather, it is likely that the high DIC and carbonate content of these systems provide a non‐limiting carbon source allowing for expression of large photosynthetic offsets, in contrast to the smaller offsets observed in saline, organic‐rich and hot spring microbial mats.     

Effects of long-term chlorimuron-ethyl application on the diversity and antifungal activity of soil Pseudomonas spp. in a soybean field in Northeast China

The herbicide chlorimuron-ethyl has been applied widely for weed control in farmland, especially in soybean fields in China over the past decade, but the chronic effects of this herbicide on soil microorganisms, particularly Pseudomonas spp., is not well understood. Taking a continuously cropped soybean field in the town of Fuyuan—a soybean production base of Heilongjiang Province in Northeast China—as a case study, soil samples were collected from plots having received 0-, 5-, and 10-year applications of chlorimuron-ethyl (30 g active component of chlorimuron-ethyl/ha/year) to study the abundance and diversity of Pseudomonas spp. Meanwhile, an in vitro assay was used to examine the antifungal activities of isolated Pseudomonas spp. against soil-borne pathogens (Fusarium graminearum, Fusarium oxysporum, and Rhizoctonia solani) causing soybean root rot disease. The production of siderophore, hydrogen cyanide (HCN), and lytic enzymes (cellulase, pectinase, and chitinase) by Pseudomonas spp. was also investigated. With 5- and 10- year chlorimuron-ethyl application, the numbers of soil Pseudomonas spp. decreased from 121?×?10² CFU/g dry soil in the control to 40?×?10² CFU/g dry soil and 13?×?10² CFU/g dry soil, and the Shannon index values decreased from 6.23 to 3.71 and 1.73, respectively. The numbers of antifungal Pseudomonas spp. also decreased, and the proportions of Pseudomonas spp. with antifungal activities against the different test pathogens altered. All the antifungal Pseudomonas spp. could produce siderophore and HCN but not lytic enzymes. The results suggest that long-term application of chlorimuron-ethyl in continuously cropped soybean field had negative effects on the abundance and diversity of soil Pseudomonas spp., including species with different antifungal activities against pathogens. Siderophore and HCN rather than lytic enzymes formed the antifungal metabolites of Pseudomonas spp., and the number of antifungal Pseudomonas that can produce siderophore and HCN decreased markedly under application of chlorimuron-ethyl, especially after 10-year application.     

Use of stable isotopes to understand food webs in Macao wetlands

In this study, components of the food-web in Macao wetlands were quantified using stable isotope ratio techniques based on carbon and nitrogen values. The δ¹³C and δ¹⁵N values of particulate organic matter (δ¹³C_POM and δ¹⁵N_POM, respectively) ranged from ?30.64 ± 1.0 to ?28.1 ± 0.7 ‰, and from ?1.11 ± 0.8 to 3.98 ± 0.7 ‰, respectively. The δ¹³C values of consumer species ranged from ?33.94 to ?16.92 ‰, showing a wide range from lower values in a freshwater lake and inner bay to higher values in a mangrove forest. The distinct dietary habits of consumer species and the location-specific food source composition were the main factors affecting the δ¹³C values. The consumer ¹⁵N-isotope enrichment values suggested that there were three trophic levels; primary, secondary, and tertiary. The primary consumer trophic level was represented by freshwater herbivorous gastropods, filter-feeding bivalves, and plankton-feeding fish, with a mean δ¹⁵N value of 5.052 ‰. The secondary consumer level included four deposit-feeding fish species distributed in Fai Chi Kei Bay and deposit-feeding gastropods in the Lotus Flower Bridge flat, with a mean δ¹⁵N value of 6.794 ‰. The tertiary consumers group consisted of four crab species, one shrimp species, and four fish species in the Lotus Flower Bridge Flat, with a mean δ¹⁵N value of 13.473 ‰. Their diet mainly comprised organic debris, bottom fauna, and rotten animal tissues. This study confirms the applicability of the isotopic approach in food web studies.     

Metabolic Quotient Measured by Free-Water Method in Six Enclosures with Different Silver Carp Densities

Quasi-continuous DO and pH measurements (total 47 days) were conducted during enclosure experiments (6 enclosures; 5 × 5 × 2.5 m), in which a biomass gradient of silver carp was created. After subtracting the air–water exchanges of O₂ and CO₂, the chemical and biochemical changes in DO (dissolved oxygen) and DIC (dissolved inorganic carbon) were estimated in order to evaluate MQ (metabolic quotient: DO change divided by DIC change) at intervals of 1 hour. By removing small absolute changes below the threshold value (0.01 mM h^?1), the averaged values of the 24 MQ means for the respective 1-hour periods ranged from 0.96 to 1.20 in the six enclosures. Because the MQs in the daytime inversely correlated well with the ratio of NH⁺ ₄–N to (NH⁺ ₄–N + NO^? ₃–N), not the ecosystems, i.e., density of fish, community structure of zooplankton and phytoplankton, but the form of nitrogen uptaken for primary production principally determined the MQs. The higher MQs observed in the daytime compared with the nighttime (from 14% to 21% except 3% for one enclosure) could not be explained by the denitrification and/or dissolution of CaCO₃ in the sediments, therefore suggesting the selectively faster decomposition of part of the organic matter provided through primary production, in other words, an accumulation of another part of the organic matter in the diurnal and/or daily time scale.     

Spatial distribution of rhizodeposit carbon of maize (Zea mays L.) in soil aggregates assessed by multiple pulse 13C labeling in the field

Background and aims

Rhizosphere effect is controlled by spatial distribution of rhizodeposits, which may be influenced by soil aggregation and soil moisture regime in relation to water uptake by roots. The objectives of this study were to measure soil organic carbon (SOC) concentration and its δ¹³C abundance by aggregate size in the rooted bulk soil and by distance in the root-free soil vertically and horizontally away from roots, and to measure DOC concentration and its δ¹³C abundance in pore water in the rooted bulk soil after a seasonal pulse labelings of ¹³CO₂ to maize (Zea mays L.).

Methods

Pulse labeling was conducted in the field once a week for 11 weeks. Soil cells (50 mm in diameter and 100 mm long) mimicking root-free soils were imbedded vertically and horizontally 25–50 mm away from the main root of a maize crop. The rooted bulk soils were sampled to extract soil pore water at different suctions and to fractionate aggregates by wet sieving. The root-free soil cells were sliced by 1 mm intervals from the root end to 20 mm away. All the sampling was 12 days after the last labeling after the crop was harvested.

Results and discussion

The δ¹³C abundance before and after the continuous labeling was ?24.20?±?0.05?‰ and ?23.80?±?0.05?‰ in the rooted bulk soil. The labeling caused increases in δ¹³C abundance in all the aggregates in the rooted bulk soil and down to 14 mm away from the roots in both the root-free cells. The δ¹³C abundance was enriched in the >2 mm and 1–2 mm aggregates (?23.17?±?0.12?‰ and ?23.26?±?0.05?‰) though the SOC concentration was not different among the >0.25 mm aggregates, indicating that rhizodeposits or their metabolites were protected and distributed widely in whole soil through soil aggregation. The δ¹³C abundance in pore water (?24.0?±?0.01?‰) was much lower than those soil aggregates and greatest from the >2 μm soil pores though the DOC concentration was greater from the <20 μm soil pores. The δ¹³C abundance was in general greater in the horizontal cell than in the vertical cell. The δ¹³C abundance decreased with the increasing distance to the roots in the vertical cell and peaked at the 5 and 6 mm distance to the roots in the horizontal cell (?23.66?±?0.11?‰ and ?23.5?±?0.10?‰), possibly due to the drier condition unfavorable to microbial decomposition in the horizontal cell. The higher δ¹³C abundance in the horizontal cell than in the vertical cell was accompanied by a lower SOC concentration and a lower C: N ratio within 3 mm away from the roots, suggesting a stronger priming effect due to the longer residence time of rhizodeposits in the horizontal cell than in the vertical cell.

Conclusions

Rhizodeposits or their metabolites were protected during soil aggregation and distributed to 14 mm beyond the rhizosphere in the natural soil-plant system. This extension is of significance in regulating the formation of soil structure and the priming of soil organic matter during the whole life cycle of plants, which needs further study.     

Unique Microbial Communities Inhabiting Underground Seawater in an Intertidal Area Utilized for Industrialized Aquaculture,as Compared with the Coastal Water

In recent decades, the decline of coastal water quality has promoted the birth of a new industrialized aquaculture mode in China, which involves the cultivation of organisms using underground seawater extracted from various depths below the intertidal zone. In view of the special physicochemical characteristics of underground seawater, the microbial community in this environment has attracted interest. In this study, the microbial community in the underground seawater of an intertidal area of the Qingdao coast of China was investigated. Compared with the upper coastal water, the underground seawater displayed lower numbers of microorganisms (2.7?±?0.3?×?10⁵ cells mL^?1 in underground seawater vs. 5.3?±?0.4?×?10⁵ cells mL^?1 in upper coastal seawater) but displayed much higher microbial diversity. At the phyla level, Proteobacteria, Bacteroidetes, Cyanobacteria, and Actinobacteria inhabited both environments, whereas bacteria in the phyla Planctomycetes, Deferribacteres, and Nitrospirae were recovered only from the underground seawater. Eighty-nine percent of the OTUs in the underground seawater were environmental specific. Furthermore, compared with coastal water, underground seawater displayed significant lower (p?<?0.05) concentration of NH₃-N, NO₂-N, PO₄-P, and DOC-C, and contained fewer potentially harmful pathogens (e.g., Verrucomicrobia/Opitutae) and more denitrifying bacteria (e.g., Shewanella denitrificans), thus making it more suitable for aquaculture.     

Stable isotopic (δ13C and δ18O) signatures of biogenic calcretes marking discontinuity surfaces: a case study from Upper Cretaceous carbonates of central Dalmatia and eastern Istria,Croatia

Biogenic calcretes associated with a regional Cretaceous to Paleogene subaerial unconformity and an intraformational composite (polygenic) surface in Upper Cretaceous intra-platform peritidal successions in central Dalmatia and eastern Istria, Croatia (Adriatic-Dinaridic Carbonate Platform), were analyzed for their δ¹³C and δ¹⁸O signatures in order to provide insight into the conditions of subaerial exposure and calcrete development. The distinctly negative δ¹³C signatures of biogenic calcretes marking the regional subaerial unconformity differ considerably from the δ¹³C values of the host marine limestones. This indicates carbon isotope exchange of primary marine CaCO₃ with CO₂ released by root and rhizomicrobial respiration and subsequent precipitation of pedogenic calcrete. The range of δ¹³C (from ?13.1 to ?8.2 ‰ Vienna PeeDee Belemnite standard, VPDB) and δ¹⁸O (from ?10.1 to ?6.1 ‰ VPDB) values of calcretes are similar to those reported from calcretes elsewhere, and the δ¹³C values of biogenic calcretes with typical Microcodium aggregates (?13.1 to ?12.3 ‰ VPDB) at the ?ibenik locality are very close to, or at the lower limit of, values for soil carbonates formed in isotopic equilibrium with soil CO₂. These values are expected for authigenic pedogenic carbonates formed under the influence of C₃ plant communities, without influence from heavier carbon from pre-existing carbonate and lack of input of atmospheric CO₂. Such low δ¹³C values support the interpretation of Microcodium aggregates as being precipitated under a direct biological control within the soil, although the relationship between formation mechanisms and stable isotope signatures of Microcodium needs further investigation. The δ¹³C values (?4.4 to ?3.6 ‰ VPDB) of rhizogenic calcretes formed inside firmground Thalassinoides burrows of the composite surface at the ?ibenik locality are more negative than the δ¹³C values of the host marine limestones, which confirms that the composite surface went through a phase of meteoric pedo(dia)genesis. However, the overall δ¹³C values of calcretes are less negative than expected, which might reflect contamination from associated primary marine carbonate. This study represents the first detailed stable isotope investigation of calcretes from carbonate successions of the External Dinarides, and the results may be applied to discontinuities present in other shallow-water carbonate rock successions.     

The relative contributions of biological and abiotic processes to carbon dynamics in subarctic sea ice

Knowledge on the relative effects of biological activity and precipitation/dissolution of calcium carbonate (CaCO₃) in influencing the air-ice CO₂ exchange in sea-ice-covered season is currently lacking. Furthermore, the spatial and temporal occurrence of CaCO₃ and other biogeochemical parameters in sea ice are still not well described. Here we investigated autotrophic and heterotrophic activity as well as the precipitation/dissolution of CaCO₃ in subarctic sea ice in South West Greenland. Integrated over the entire ice season (71 days), the sea ice was net autotrophic with a net carbon fixation of 56 mg C m^?2, derived from a sea-ice-related gross primary production of 153 mg C m^?2 and a bacterial carbon demand of 97 mg C m^?2. Primary production contributed only marginally to the TCO₂ depletion of the sea ice (7–25 %), which was mainly controlled by physical export by brine drainage and CaCO₃ precipitation. The net biological production could only explain 4 % of this sea-ice-driven CO₂ uptake. Abiotic processes contributed to an air-sea CO₂ uptake of 1.5 mmol m^?2 sea ice day^?1, and dissolution of CaCO₃ increased the air-sea CO₂ uptake by 36 % compared to a theoretical estimate of melting CaCO₃-free sea ice. There was a considerable spatial and temporal variability of CaCO₃ and the other biogeochemical parameters measured (dissolved organic and inorganic nutrients).