Effects of Carbon Source, Carbon Concentration, and Chlorination on Growth Related Parameters of Heterotrophic Biofilm Bacteria

Abstract To investigate growth of heterotrophic biofilm bacteria, a model biofilm reactor was developed to simulate a drinking water distribution system. Controlled addition of three different carbon sources (amino acids, carbohydrates, and humics) at three different concentrations (500, 1,000, and 2,000 ppb carbon) in the presence and absence of chlorine were used in separate experiments. An additional experiment was run with a 1:1:2 mixture of the above carbon sources. Biofilm and effluent total and culturable cells in addition to total and dissolved organic carbon were measured in order to estimate specific growth rates (SGRs), observed yields, population densities, and bacterial carbon production rates. Bacterial carbon production rates (μg C/L day) were extremely high in the control biofilm communities (range = 295–1,738). Both growth rate and yield decreased with increasing carbon concentrations. Therefore, biofilm growth rates were zero-order with respect to the carbon concentrations used in these experiments. There was no correlation between growth rate and carbon concentration, but there was a significant negative correlation between growth rate and biofilm cell density (r=−0.637, p= 0.001 control and r=−0.57, p= 0.021 chlorinated biofilms). Growth efficiency was highest at the lowest carbon concentration (range = 12–4.5%, amino acids and humics respectively). Doubling times ranged from 2.3–15.4 days in the control biofilms and 1–12.3 days in the chlorinated biofilms. Growth rates were significantly higher in the presence of chlorine for the carbohydrates, humics, and mixed carbon sources (p= 0.004, < 0.0005, 0.013, respectively). The concept of r/K selection theory was used to explain the results with respect to specific growth rates and yields. Humic removal by the biofilm bacteria (78% and 56% for the control and chlorinated biofilms, respectively) was higher than previously reported literature values for planktonic bacteria. A number of control experiments indicated that filtration of drinking water was as effective as chlorination in controlling bacterial biofilm growth. Received: 26 March 1999; Accepted: 3 August 1999; Online Publication: 15 February 2000     

草地农业生态系统的碳平衡分析方法

根据草地农业生态系统的结构,它的碳平衡为4个生产层的碳平衡之和,也是3个界面的碳平衡之和,而某一生产层或者某一界面的碳平衡则是其固定、输入、排放和输出的碳之和。草地农业生态系统4个生产层的碳平衡分析方法定量重要生产环节的碳汇与碳源过程,便于草业生产改进碳汇管理;草地农业生态系统3个界面的碳平衡分析方法显示碳源和碳汇的发生机理,及其空间和数量关系,便于调控草业生产组分以增汇减排;但是,这两个方法不易区分碳的来源和去向,难以明确其利用效率。草地农业生态系统碳平衡分析的输入/输出法定量地指示碳的来源和去向,以及碳效率,计算简单,但是较为概括,不利于牧场尺度的草业碳汇管理。以中国祁连山甘肃马鹿牧场和澳大利亚塔斯玛尼亚奶牛牧场为例,用3种方法分析了两个牧场的碳平衡,结果表明,放牧管理的草业系统的主要碳源是休闲旅游、产品加工流通环节产生的温室气体,主要碳汇是草地和土壤中贮存的碳,好的草地管理可以增汇减排。     

Synthesis of Extracellular Proteinase by Pseudomonas fluorescens Under Conditions of Limiting Carbon, Nitrogen, and Phosphate

The influence of carbon, nitrogen, and phosphate concentrations on growth and proteinase production by Pseudomonas fluorescens 32A was examined. In mineral salts medium containing dialyzed skim milk supernatant as an inducer, maximum growth was obtained at 1.0 and 2.5 mM orthophosphate at 20 and 5°C, respectively. At both temperatures, 5 mM orthophosphate was required for maximum proteinase production, whereas significant inhibition was found at 10 mM. Orthophosphate was the only phosphate compound able to support growth. With sodium pyruvate as the carbon source, maximum enzyme synthesis was at 100 mM carbon at both temperatures. At both 20 and 5°C maximum growth and enzyme production was found with 10 mM NH₄Cl. A bioassay for available phosphate based on the growth of P. fluorescens 32A in phosphate-limited mineral salts medium showed that skim milk and skim milk supernatant contained 50 and 10 mM orthophosphate, respectively. Proteinase production in skim milk was 2.6- and 12-fold greater than that in optimal mineral salts medium at 20 and 5°C, respectively. These results suggest that proteinase production in milk does not occur as a result of nutrient limitation and may be regulated in part by milk phosphates.     

Effects of Road Decommissioning on Carbon Stocks,Losses, and Emissions in North Coastal California

During the last 3 decades, many road removal projects have been implemented on public and private lands in the United States to reduce erosion and other impacts from abandoned or unmaintained forest roads. Although effective in decreasing sediment production from roads, such activities have a carbon (C) cost as well as representing a carbon savings for an ecosystem. We assessed the carbon budget implications of 30 years of road decommissioning in Redwood National Park in north coastal California. Road restoration techniques, which evolved during the program, were associated with various carbon costs and savings. Treatment of 425 km of logging roads from 1979 to 2009 saved 72,000 megagrams (Mg) C through on‐site soil erosion prevention, revegetation, and soil development on formerly compacted roads. Carbon sequestration will increase in time as forests and soils develop more fully on the restored sites. The carbon cost for this road decommissioning work, based on heavy equipment and vehicle fuel emissions, short‐term soil loss, and clearing of vegetation, was 23,000 Mg C, resulting in a net carbon savings of 49,000 Mg C to date. Nevertheless, the degree to which soil loss is a carbon sink or source in steep mountainous watersheds needs to be further examined. The ratio of carbon costs to savings will differ by ecosystem and road removal methodology, but the procedure outlined here to assess carbon budgets on restoration sites should be transferable to other systems.     

Net carbon flux from agriculture: Carbon emissions, carbon sequestration, crop yield, and land-use change

There is a potential to sequester carbon in soil by changing agricultural management practices. These changes in agricultural management can also result in changes in fossil-fuel use, agricultural inputs, and the carbon emissions associated with fossil fuels and other inputs. Management practices that alter crop yields and land productivity can affect the amount of land used for crop production with further significant implications for both emissions and sequestration potential. Data from a 20-year agricultural experiment were used to analyze carbon sequestration, carbon emissions, crop yield, and land-use change and to estimate the impact that carbon sequestration strategies might have on the net flux of carbon to the atmosphere. Results indicate that if changes in management result in decreased crop yields, the net carbon flux can be greater under the new system, assuming that crop demand remains the same and additional lands are brought into production. Conversely, if increasing crop yields lead to land abandonment, the overall carbon savings from changes in management will be greater than when soil carbon sequestration alone is considered.     

Uncertainty and Variability in Product Carbon Footprinting

Recent years have seen increasing interest in life cycle greenhouse gas emissions accounting, also known as carbon footprinting, due to drivers such as transportation fuels policy and climate‐related eco‐labels, sometimes called carbon labels. However, it remains unclear whether applications of greenhouse gas accounting, such as carbon labels, are supportable given the level of precision that is possible with current methodology and data. The goal of this work is to further the understanding of quantitative uncertainty assessment in carbon footprinting through a case study of a rackmount electronic server. Production phase uncertainty was found to be moderate (±15%), though with a high likelihood of being significantly underestimated given the limitations in available data for assessing uncertainty associated with temporal variability and technological specificity. Individual components or subassemblies showed varying levels of uncertainty due to differences in parameter uncertainty (i.e., agreement between data sets) and variability between production or use regions. The use phase displayed a considerably higher uncertainty (±50%) than production due to uncertainty in the useful lifetime of the server, variability in electricity mixes in different market regions, and use profile uncertainty. Overall model uncertainty was found to be ±35% for the whole life cycle, a substantial amount given that the method is already being used to set policy and make comparative environmental product declarations. Future work should continue to combine the increasing volume of available data to ensure consistency and maximize the credibility of the methods of life cycle assessment (LCA) and carbon footprinting. However, for some energy‐using products it may make more sense to increase focus on energy efficiency and use phase emissions reductions rather than attempting to quantify and reduce the uncertainty of the relatively small production phase.     

Organic Carbon Flux to a Large Salt Lake: Pyramid Lake,Nevada, USA

Energy flux to a large, deep, salt lake from phytoplankton, periphyton and macrophyte primary production as well as fluvial transport and wind-transported terrestrial vegetation and dust were quantified. Average areal phytoplankton net photosynthesis was 511 mg C m⁻² d⁻¹. Highest rates were during water-blooms of the bluegreen alga, Nodularia spumigena. Although areal daily net photosynthesis by periphyton in Pyramid Lake was comparable to other salt lakes, annual carbon influx by periphyton was small due to the lake's graben morphology and moderate euphotic depth (mean, 11.9 m). Macrophytes were uncommon and, therefore a minor source of energy. Truckee River is the only major fluvial discharge to Pyramid Lake and dissolved organic carbon was the principal organic carbon fraction in river water. Large upstream water diversions coupled with several drought years resulted in an average fluvial organic carbon load of only 7.3 g Cm⁻²y⁻¹ or 4% of median phytoplankton net photosynthesis. Tumbleweeds were the most common terrestrial plant material observed in Pyramid Lake comprising a maximum projected importance of 6% of total annual carbon input. Windborne dust represented < .1% of annual carbon input. Phytoplankton primary production is the predominant energy source to Pyramid Lake, accounting for over 80% of annual carbon influx. The relative magnitude of autochthonous and allochthonous vectors to the annual carbon budget of this desert salt lake are comparable to those of the few other large lakes for which detailed energy input budgets have been calculated.     

Marine Heterotrophic Bacteria in Continuous Culture,the Bacterial Carbon Growth Efficiency,and Mineralization at Excess Substrate and Different Temperatures

To model the physiological potential of marine heterotrophic bacteria, their role in the food web, and in the biogeochemical carbon cycle, we need to know their growth efficiency response within a matrix of different temperatures and degrees of organic substrate limitation. In this work, we present one part of this matrix, the carbon growth efficiencies of marine bacteria under different temperatures and nonlimiting organic and inorganic substrate supply. We ran aerobic turbidostats with glucose enriched seawater, inoculated with natural populations of heterotrophic marine bacteria at 10, 14, 18, 22, and 26°C. The average cell-specific growth rates increased with temperature from 1.17 to 2.6 h−1. At steady-state total CO₂ production, biomass production [particulate organic carbon (POC) and nitrogen (PON)], and viruslike particle abundance was measured. CO₂ production and specific growth rate increased with increasing temperature. Bacterial carbon growth efficiency (BCGE), the particulate carbon produced per dissolved carbon utilized, varied between 0.12 and 0.70. Maximum BCGE values and decreased specific respiration rates occurred at higher temperatures (22 and 26°C) and growth rates. This trend was largely attributable to an increase in POC per cell abundance; when the BCGE was recalculated, parameterizing the biomass as the product of cell concentration and a constant cellular carbon content, the opposite trend was observed.     

Is Net Ecosystem Production Equal to Ecosystem Carbon Accumulation?

Net ecosystem production (NEP), defined as the difference between gross primary production and total ecosystem respiration, represents the total amount of organic carbon in an ecosystem available for storage, export as organic carbon, or nonbiological oxidation to carbon dioxide through fire or ultraviolet oxidation. In some of the recent literature, especially that on terrestrial ecosystems, NEP has been redefined as the rate of organic carbon accumulation in the system. Here we argue that retaining the original definition maintains the conceptual coherence between NEP and net primary production and that it is congruous with the widely accepted definitions of ecosystem autotrophy and heterotrophy. Careful evaluation of NEP highlights the various potential fates of nonrespired carbon in an ecosystem.     

Carbon Source and Micronutrient Requirements of the Aquatic Phycomycete, Catenaria anguillulae Sorokin

The requirements of the aquatic Phycomycete, Catenaria anguillulaewere analysed in liquid, shake cultures using a standardizedzoospore inoculum. Growth was determined by measuring mycelialdry weight and rate of production of titratable acid. D-glucose was the best carbon source and had an optimum concentrationof 166 mM of carbon for the medium used. When other carbon sourceswere supplied, only those related to glucose (fructose and mannose)or composed of glucose units with an alpha-linkage (maltose,glycogen, and starch) were readily utilized. Lactic acid wasdetermined qualitatively as an end-product of carbon metabolism. The optimum level of phosphate was 1.0 mM. The optimum concentrationof EDTA was 0.032 mM. Of the chelated cations included in themedium only the omission of iron, zinc, calcium, or magnesiumreduced growth. Concentrations of calcium below 0.4 mM and ofmagnesium below 0.2 mM were limiting; whereas, concentrationsof both ions up to 1 mM were non-toxic.     

Carbon source for reproduction in a spring ephemeral herb, Corydalis ambigua (Papaveraceae)

1.  The carbon source for reproduction in plants may differ between flowering and fruiting stages. To clarify how spring ephemerals use current photosynthetic products for reproduction, the allocation patterns of photosynthate at flowering and fruiting and the effects of resource limitation on reproductive performance in Corydalis ambigua were assessed.
2.  A ¹³C tracing experiment revealed that about 20% of the current photosynthetic carbon was used for reproduction at both flowering and fruiting. The proportion of ¹³C allocated to fruits was constant irrespective of the light level. In contrast, ¹³C translocation to tubers increased at fruiting, and this trend was accelerated when plants were shaded.
3.  Defoliation treatment significantly reduced nectar production and tuber mass, while seed production was not affected. Therefore, when carbon assimilation was limited, carbon was preferentially allocated to current reproduction (seeds) rather than to pollinator attraction (nectar) or storage (tuber).
4.  If seed production is partly supported by carbohydrate reserved in the old tissue of tubers, nectar and seed production may not compete strongly for carbon sources. In contrast to the ability of high seed production, the susceptibility of nectar production to current photosynthesis indicates that seed production of this species is basically limited by pollen capture.
5.  Therefore, temporal separation of resource pool for reproduction may mitigate the joint limitation of seed production between pollinator attraction and resource availability. Temporal variation of the sink–source balance of storage organ is crucial to understand the cost of reproduction in perennial plants.     

Carbon allocation in forest ecosystems

Carbon allocation plays a critical role in forest ecosystem carbon cycling. We reviewed existing literature and compiled annual carbon budgets for forest ecosystems to test a series of hypotheses addressing the patterns, plasticity, and limits of three components of allocation: biomass, the amount of material present; flux, the flow of carbon to a component per unit time; and partitioning, the fraction of gross primary productivity (GPP) used by a component. Can annual carbon flux and partitioning be inferred from biomass? Our survey revealed that biomass was poorly related to carbon flux and to partitioning of photosynthetically derived carbon, and should not be used to infer either. Are component fluxes correlated? Carbon fluxes to foliage, wood, and belowground production and respiration all increased linearly with increasing GPP (a rising tide lifts all boats). Autotrophic respiration was strongly linked to production for foliage, wood and roots, and aboveground net primary productivity and total belowground carbon flux (TBCF) were positively correlated across a broad productivity gradient. How does carbon partitioning respond to variability in resources and environment? Within sites, partitioning to aboveground wood production and TBCF responded to changes in stand age and resource availability, but not to competition (tree density). Increasing resource supply and stand age, with one exception, resulted in increased partitioning to aboveground wood production and decreased partitioning to TBCF. Partitioning to foliage production was much less sensitive to changes in resources and environment. Overall, changes in partitioning within a site in response to resource supply and age were small (<15% of GPP), but much greater than those inferred from global relationships. Across all sites, foliage production plus respiration, and total autotrophic respiration appear to use relatively constant fractions of GPP – partitioning to both was conservative across a broad range of GPP – but values did vary across sites. Partitioning to aboveground wood production and to TBCF were the most variable – conditions that favored high GPP increased partitioning to aboveground wood production and decreased partitioning to TBCF. Do priorities exist for the products of photosynthesis? The available data do not support the concept of priorities for the products of photosynthesis, because increasing GPP increased all fluxes. All facets of carbon allocation are important to understanding carbon cycling in forest ecosystems. Terrestrial ecosystem models require information on partitioning, yet we found few studies that measured all components of the carbon budget to allow estimation of partitioning coefficients. Future studies that measure complete annual carbon budgets contribute the most to understanding carbon allocation.     

Carbon and energy balances of glucose fermentation with hydrogenproducing bacterium Citrobacter amalonaticus Y19

For the newly isolated H2-producing chemoheterotrophic bacterium Citrobacter amalonaticus Y19, anaerobic glucose metabolism was studied in batch cultivation at varying initial glucose concentrations (3.5- 9.5 g/l). The carbon-mass and energy balances were determined and utilized to analyze the carbon metabolic-pathways network. The analyses revealed (a) variable production of major metabolites (H2, ethanol, acetate, lactate, CO2, and cell mass) depending on initial glucose levels; (b) influence of NADH regeneration on the production of acetate, lactate, and ethanol; and (c) influence of the molar production of ATP on the production of biomass. The results reported in this paper suggest how the carbon metabolic pathway(s) should be designed for optimal H2 production, especially at high glucose concentrations, such as by blocking the carbon flux via lactate dehydrogenase from the pyruvate node.     

Carbon and nitrogen source nutrition of fumagillin biosynthesis by Aspergillus fumigatus

The carbon and nitrogen source requirements of Aspergillus fumigatus NRRL 2436 for growth and production of the angiogenesis inhibitor fumagillin were studied in chemically defined media. Both carbon and nitrogen sources strongly influenced fumagillin formation. Two out of 29 carbon sources tested interfered with fumagillin biosynthesis. The best combination of two carbon sources was 30 g L(-1) xylan and 50 g L(-1) mannose. Of fifteen nitrogen sources tested, three ammonium salts (chloride, sulfate, and dibasic phosphate) failed to support fumagillin formation, presumably due to the low pH which developed. The dosage-response study of the best nitrogen source, L-glutamic acid, revealed that 9 g L(-1) was optimal. Volumetric production of fumagillin was increased by 15-fold over that in the starting (Peterson-Goldstein) medium as a result of these findings.     

Carbon mass balance evaluation of cellulase production on soluble and insoluble substrates

A methodology is described and applied for performing carbon mass balances across cellulase enzyme production processes using both soluble sugar and insoluble cellulose substrates. The fungus Trichoderma reesei was grown on either glucose, lactose, or cellulose in aerobic batch mode, and the evolution of the main carbonaceous components (cell mass, cellulose, soluble protein, adsorbed protein, sugars, and carbon dioxide) was followed. A variety of analytical techniques were utilized to measure these components, including (i) gravimetric analysis, (ii) near-infrared spectroscopy, (iii) bicinchoninic acid based soluble protein measurement, (iv) gas mass spectrometry and flow rate, (v) CHNS/O elemental analyses, and (vi) high-performance liquid chromatography. The combined set of measurements allowed carbon mass balances across the cellulase production process to be assessed to determine the consistency of the underlying kinetic data. Results demonstrate the capability to determine the levels and distribution of all major carbonaceous components during the cellulase production process on both soluble and insoluble substrates. Average carbon mass balance closures were near 100% during early stages (<72 h) of the cultivations using glucose, lactose, or cellulose as the substrates, but carbon mass closures trended high later in the cultivation. Analysis of carbon allocation results suggests that an error in the gas mass flow rate measurement was the primary cause for carbon mass balance closures to exceed 110% late in the process.     

小兴安岭谷地云冷杉林的碳密度与生产力

采用样地清查和异速生长方程法,量化了处于衰退状态的小兴安岭谷地云冷杉林的森林碳密度和生产力.结果表明： 2011年森林碳密度总量为268.14 t C·hm^-2,其中植被碳密度、碎屑碳密度和土壤碳密度分别为74.25、16.86和177.03 t C·hm^-2.2006—2011年,乔木层碳密度从80.86 t C·hm^-2减少到71.73 t C·hm^-2,主要树种冷杉、白桦、云杉和兴安落叶松的碳密度年均减少比例分别为0.5%、1.2%、2.7%和3.7%,毛赤杨、红松和花楷槭的碳密度年均增加比例分别为2.9%、3.9%和7.2%.森林净初级生产力(NPP)为4.69 t C·hm^-2·a^-1,地下部和地上部NPP比值为0.56,凋落物损失部分是总NPP的最大组分,所占比例为34.5%.森林生态系统中2个主要碳输出途径异养呼吸和粗木质残体分解的年通量分别为293.67和119.29 g C·m^-2·a^-1.森林净生态系统生产力(NEP)为55.90 g C·m^-2·a^-1.研究结果表明,处于衰退状态的谷地云冷杉林仍具有一定的碳汇功能.     

CRP-Mediated Carbon Catabolite Regulation of Yersinia pestis Biofilm Formation Is Enhanced by the Carbon Storage Regulator Protein,CsrA

The natural transmission of Yersinia pestis is reliant upon biofilm blockage of the flea vector. However, the environmentally-responsive adaptive regulators which facilitate Y. pestis biofilm production in accordance with the flea midgut milieu are not well understood. We seek to establish the impact of available carbon source metabolism and storage upon Y. pestis biofilm production. Our findings demonstrate that Y. pestis biofilm production is subject to carbon catabolite regulation in which the presence of glucose impairs biofilm production; whereas, the sole metabolism of alternate carbon sources promotes robust biofilm formation. This observation is facilitated by the cAMP receptor protein, CRP. In accordance with a stark growth defect, deletion of crp in both CO92 and KIM6+ Y. pestis strains significantly impaired biofilm production when solely utilizing alternate carbon sources. Media supplementation with cAMP, a small-molecule activator of CRP, did not significantly alter Y. pestis biofilm production. Furthermore, CRP did not alter mRNA abundance of previously-characterized hms biofilm synthesis and regulation factors. Therefore, our findings indicate CRP does not confer a direct stimulatory effect, but may indirectly promote Y. pestis biofilm production by facilitating the alternate carbon source expression profile. Additionally, we assessed the impact of the carbon storage regulator protein, CsrA, upon Y. pestis biofilm production. Contrary to what has been described for E. coli, Y. pestis biofilm formation was found to be enhanced by CsrA. Regardless of media composition and available carbon source, deletion of csrA significantly impaired Y. pestis biofilm production. CsrA was found to promote Y. pestis biofilm production independent of glycogen regulation. Loss of csrA did not significantly alter relative hmsH, hmsP, or hmsT mRNA abundance. However, deletion of hmsP in the csrA-deficient mutant enabled excessive biofilm production, suggesting CsrA enables potent Y. pestis biofilm production through cyclic diguanylate regulation.     

碳源、氮源对异养单针藻Monoraphidium sp. FXY-10油脂积累和脂肪酸组成的影响

研究了碳源与氮源对单针藻Monoraphidium sp. FXY-10异养培养的影响。以BG-11为基础培养基,通过添加不同类型、浓度梯度碳源和氮源,比较分析微藻生物量、油脂积累以及脂肪酸组成。结果表明,以葡萄糖作碳源,硝酸钠为氮源,微藻细胞积累的油脂是理想的生物柴油制备原料。硝酸钠浓度分别为1.00、3.00和5.00 g/L时,对油脂产量影响不显著(P>0.05)。葡萄糖浓度为10.00 g/L,硝酸钠为氮源油脂产量达到实验最高值0.84 g/L,其油脂脂肪酸组成主要由C16:0和C18:1等短链饱和脂肪酸和单不饱和脂肪酸组成,不饱和度值(DU)为61.98,相对偏低。     

Landscape-Scale Simulation of Heterogeneous Fire Effects on Pyrogenic Carbon Emissions,Tree Mortality,and Net Ecosystem Production

Fire influences carbon dynamics from local to global scales, but many uncertainties remain regarding the remote detection and simulation of heterogeneous fire effects. This study integrates Landsat-based remote sensing and Biome-BGC process modeling to simulate the effects of high-, moderate-, and low-severity fire on pyrogenic emissions, tree mortality, and net ecosystem production. The simulation area (244,600 ha) encompasses four fires that burned approximately 50,000 ha in 2002–2003 across the Metolius Watershed, Oregon, USA, as well as in situ measurements of postfire carbon pools and fluxes that we use for model evaluation. Simulated total pyrogenic emissions were 0.732 Tg C (2.4% of equivalent statewide anthropogenic carbon emissions over the same 2-year period). The simulated total carbon transfer due to tree mortality was fourfold higher than pyrogenic carbon emissions, but dead wood decomposition will occur over decades. Immediately postfire, burned areas were a simulated carbon source (net C exchange: −0.076 Tg C y⁻¹; mean ± SD: −142 ± 121 g C m⁻² y⁻¹). As expected, high-severity, stand-replacement fire had disproportionate carbon impacts. The per-unit area effects of moderate-severity fire were substantial, however, and the extent of low-severity fire merits its inclusion in landscape-scale analyses. These results demonstrate the potential to reduce uncertainties in landscape to regional carbon budgets by leveraging Landsat-based fire products that account for both stand-replacement and partial disturbance.