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Zero net growth in a membrane bioreactor with complete sludge retention.

A bench-scale membrane bioreactor was operated with complete sludge retention in order to evaluate biological processes and biomass characteristics over the long term. The investigation was carried out by feeding a bench-scale plant with real sewage under constant volumetric loading rate (VLR = 1.2 gCOD L(react)(-1) h(-1)). Biological processes were monitored by measuring substrate removal efficiencies and biomass-related parameters. The latter included bacterial activity as determined through respirometric tests specifically aimed at investigating long term heterotrophic and nitrifying activity. After about 180 days under the imposed operating conditions, the system reached equilibrium conditions with constant VSS concentration of 16-18gL(-1), organic loading rate (OLR) below 0.1 gCOD gVSS(-1) d(-1) and specific respiration rates of 2-3 mgO2 gVSS(-1) h(-1). These conditions were maintained for more than 150 days, confirming that an equilibrium had been achieved between biomass growth, endogenous metabolism, and solubilization of inorganic materials.

Acetates↗

Determining the domestic specific loads of two wastewater plants of the Paris conurbation (France) with contrasted treatments: a step for exploring the effects of the application of the European Directive.

The effluents of wastewater treatment plants (WTTP) discharged into the rivers considerably affect the biogeochemical functioning of the system. In this paper, we characterize both raw and treated domestic wastewater from two WTTPs of Parisian agglomeration using different process treatments (Achères WWTP with a secondary treatment and Colombes WWTP with a tertiary one). In addition to the classical variables, we analyse the input of bacteria, both the heterotrophs and the nitrifyers. Tertiary treatment leads to significantly decrease ammonium-specific load (< 2 g KjN inhab equ.(-1) instead of 9 g KjN inhab equ.(-1) for secondary treatment) and notably reduces the one of organic matter (approximately 2.5 g biological oxygen demand (BOD) inhab equ.(-1) instead of approximately 7.5 g BOD inhab equ.(-1) for secondary treatment); it is therefore promising to improve oxygen status of both the Seine river and its estuary. In terms of total bacterial biomass abatement (the heterotrophs mostly), bioreactors (at Colombes WWTP) eliminate 12% more bacterial biomass than the activated sludge treatment (at Achères WWTP). Regarding the nitrifying bacteria, a tertiary treatment in bioreactors eliminates reverse similar 90% of both nitrifying bacteria and nitrogen pollution of wastewater. Bacterial populations are characterized by large size bacteria (> 1 microm) with a higher growth rate, that represent in the treatment plant effluents 70% of the biomass. These large size bacteria have therefore a strong impact in the organic matter degradation and oxygen consumption. Relationships between classical physical-chemical variables routinely analysed in WWTPs laboratory and bacterial biomass (heterotrophic and nitrifying) are established, in order to quantify the ecological role of the allochthonous bacteria brought into the river system. In addition, domestic specific loads are calculated for both raw and treated effluents of the two types of WWTPs. As the application of the European Water Directive requires to upgrade the wastewater treatment at Achères WWTP as soon as 2007 for 90% nitrification and 30% denitrification and in 2015 for further denitrification (up to 70%), the results of this study can be taken as point-source constraints into the modelling approach already developed for the Seine basin, and chosen to test the implementation of the Water Frame Directive.

Bacteria↗

Climate, energy and diversity.

In recent years, a number of species-energy hypotheses have been developed to explain global patterns in plant and animal diversity. These hypotheses frequently fail to distinguish between fundamentally different forms of energy which influence diversity in dissimilar ways. Photosynthetically active radiation (PAR) can be utilized only by plants, though their abundance and growth rate is also greatly influenced by water. The Gibbs free energy (chemical energy) retained in the reduced organic compounds of tissue can be utilized by all heterotrophic organisms. Neither PAR nor chemical energy influences diversity directly. Both, however, influence biomass and/or abundance; diversity may then increase as a result of secondary population dynamic or evolutionary processes. Temperature is not a form of energy, though it is often used loosely by ecologists as a proxy for energy; it does, however, influence the rate of utilization of chemical energy by organisms. It may also influence diversity by allowing a greater range of energetic lifestyles at warmer temperatures (the metabolic niche hypothesis). We conclude that there is no single species/energy mechanism; fundamentally different processes link energy to abundance in plants and animals, and diversity is affected secondarily. If we are to make progress in elucidating these mechanisms, it is important to distinguish climatic effects on species' distribution and abundance from processes linking energy supply to plant and animal diversity.

Animals↗

Bacteria in bivalve shellfish with special reference to the oyster.

The bacterial flora of the Pacific oyster Crassostrea gigas, the sea mussel Perna viridis and the arkshell clam Scapharca cornea differed considerably from that of seawater in both numbers and generic composition. The numbers of heterotrophic bacteria in the bivalve shellfish, including the anaerobes and spore-forming bacteria, were greater than that in the surrounding water. Pseudomonas spp. were the dominant organisms, comprising over one third of the 321 strains characterized after isolation from the bivalves and seawater. Other bacteria isolated from the shellfish included Vibrio, Acinetobacter, and Aeromonas spp., whereas the seawater flora consisted mainly of coliform organisms, coryneform bacteria and Flavobacterium/Cytophaga spp. Bacteria associated with the deposit-feeding clams were higher in density and more distinct in generic composition as compared with those in the suspension-feeding oysters and mussels. Over 90% of the coliform and heterotrophic bacteria in oysters were found in organs associated with the digestive tract. Coliforms were mainly found in the stomach while heterotrophs were present in both stomach and the lower intestine. The results suggest that the stomach flora of oysters are mainly derived from the external environment and, through a process of selection and multiplication, that it may be gradually replaced by a more indigenous population which dominates the lower digestive tract.

Animals↗

Use of a fluorescent redox probe for direct visualization of actively respiring bacteria.

The redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) was employed for direct epifluorescent microscopic enumeration of respiring bacteria in environmental samples. Oxidized CTC is nearly colorless and is nonfluorescent; however, the compound is readily reduced via electron transport activity to fluorescent, insoluble CTC-formazan, which accumulates intracellularly. Bacteria containing CTC-formazan were visualized by epifluorescence microscopy in wet-mount preparations, on polycarbonate membrane filter surfaces, or in biofilms associated with optically opaque surfaces. Counterstaining of CTC-treated samples with the DNA-specific fluorochrome 4',6-diamidino-2-phenylindole allowed enumeration of active and total bacterial subpopulations within the same preparation. Municipal wastewater, groundwater, and seawater samples supplied with exogenous nutrients yielded CTC counts that were generally lower than total 4',6-diamidino-2-phenylindole counts but typically equal to or greater than standard heterotrophic (aerobic) plate counts. In unsupplemented water samples, CTC counts were typically lower than those obtained with the heterotrophic plate count method. Reduction of CTC by planktonic or biofilm-associated bacteria was suppressed by formaldehyde, presumably because of inhibition of electron transport activity and other metabolic processes. Because of their bright red fluorescence (emission maximum, 602 nm), actively respiring bacteria were readily distinguishable from abiotic particles and other background substances, which typically fluoresced at shorter wavelengths. The use of CTC greatly facilitated microscopic detection and enumeration of metabolically active (i.e., respiring) bacteria in environmental samples.

Bacteria↗

A generalized model for aerobic granule-based sequencing batch reactor. 1. Model development.

A generalized model was established for simulating an aerobic granule-based sequencing batch reactor (SBR) with considerations of biological processes, reactor hydrodynamics, mass transfer, and diffusion. Methodology of discretization was effectively used forthe model development and calculations. The activated sludge model no.1 was modified to describe the biological processes within the granules. Based on the difference between the calculated and measured results, the model structure was further improved through introducing simultaneous consumption of soluble substrates by storage and heterotrophs growth with a changeable reaction rate. Model calculations were conducted using a MATLAB program. The calculation results show the respective contributions of granules in different size fractions and slices to the overall change of model component concentrations. Moreover, oxygen concentration profiles within granules and oxygen consumption rate varied in one operating cycle. This confirms the applicability and validity of the discretization method and the model structure.

Aerobiosis↗

Representative freshwater bacterioplankton isolated from Crater Lake, Oregon.

High-throughput culturing (HTC) methods that rely on dilution to extinction in very-low-nutrient media were used to obtain bacterial isolates from Crater Lake, Oregon. 16S rRNA sequence determination and phylogenetic reconstruction were used to determine the potential ecological significance of isolated bacteria, both in Crater Lake and globally. Fifty-five Crater Lake isolates yielded 16 different 16S rRNA gene sequences. Thirty of 55 (55%) Crater Lake isolates had 16S rRNA gene sequences with 97% or greater similarity to sequences recovered previously from Crater Lake 16S rRNA gene clone libraries. Furthermore, 36 of 55 (65%) Crater Lake isolates were found to be members of widely distributed freshwater groups. These results confirm that HTC is a significant improvement over traditional isolation techniques that tend to enrich for microorganisms that do not predominate in their environment and rarely correlate with 16S rRNA gene clone library sequences. Although all isolates were obtained under dark, heterotrophic growth conditions, 2 of the 16 different groups showed evidence of photosynthetic capability as assessed by the presence of puf operon sequences, suggesting that photoheterotrophy may be a significant process in this oligotrophic, freshwater habitat.

Actinobacteria↗

[Microbial utilization of mixed substrates].

Decomposition of substrates by heterotrophic microorganisms is accomplished in natural biotopes such as in soil and in waters, on or in macroorganisms but also in laboratory and industrial biotopes. The interest of man in these processes is manifold. Starting with the division of substrates into three groups of simple substrates, complex and mixed substrates with or without solid particles their qualitative and quantitative occurrence in nature and their significance in biotechnology will be discussed. In the decomposition of these substrates their utilization by pure cultures or mixed populations is to be exactly distinguished. Simple growth curves, di- or polyauxy, sequences of decomposition of simple substrates of a mixed substrate, population changes and successions are only some of the phenomena occurring in this process. The pathways of catabolism are subjected to manifold regulations on the three levels of stoichiometric regulation, the regulation of enzyme activity and the regulation of enzyme synthesis. In natural biotopes there is hardly a constant substrate supply over a longer period. That's why certain mechanisms of regulation are permanently acting. Thus the "normal" physiological state for microorganisms is characterized by permanent transition situations--called "transients". These reactions are also applied to many biotechnological processes.

Allosteric Regulation↗

Temperature affects stoichiometry and biochemical composition of Escherichia coli.

Temperature is a master variable controlling biochemical processes in organisms, and its effects are manifested on many organizational levels in organisms and ecosystems. We examined the effects of temperature on the biochemical composition and stoichiometry of a model heterotrophic bacterium, Escherichia coli K-12, held at constant growth rate in chemostats. Increasing temperature led to increased cellular organic carbon (C) and organic nitrogen (N) with decreased phosphorus (P) content, leading to increased C/P and N/P biomass ratios. P content was related to cellular RNA, which is P-rich (9-10% by weight) and nonnucleic acid P (presumably composed of mostly phospholipids, intracellular phosphate, and polyphosphate). These results indicate that E. coli allocates an increased proportion of its P cell quota toward assembly (ribosomes) at low temperatures and an increasing proportion toward resource acquisition machinery (membranes) at higher temperatures. If these results are relevant to the behavior of prokaryotic heterotrophs in natural settings (the gut, soils, lakes, oceans, etc.), it suggests greater nutrient regeneration and less microbial nutrient retention as temperatures increase.

Biomass↗

Harmful algal blooms: causes, impacts and detection.

Blooms of autotrophic algae and some heterotrophic protists are increasingly frequent in coastal waters around the world and are collectively grouped as harmful algal blooms (HABs). Blooms of these organisms are attributed to two primary factors: natural processes such as circulation, upwelling relaxation, and river flow; and, anthropogenic loadings leading to eutrophication. Unfortunately, the latter is commonly assumed to be the primary cause of all blooms, which is not the case in many instances. Moreover, although it is generally acknowledged that occurrences of these phenomena are increasing throughout the world's oceans, the reasons for this apparent increase remain debated and include not only eutrophication but increased observation efforts in coastal zones of the world. There is a rapidly advancing monitoring effort resulting from the perception of increased impacts from these HABs, manifested as expanding routine coastal monitoring programs, rapid development and deployment of new detection methods for individual species, toxins, and toxicities, and expansion of coastal modeling activities towards observational forecasts of bloom landfall and eventually bloom prediction. Together, these many efforts will provide resource managers with the tools needed to develop effective strategies for the management and mitigation of HABs and their frequently devastating impacts on the coastal environment.

Environment↗

Diplonemid glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and prokaryote-to-eukaryote lateral gene transfer.

Lateral gene transfer refers to the movement of genetic information from one genome to another, and the integration of that foreign DNA into its new genetic environment. There are currently only a few well-supported cases of prokaryote-to-eukaryote transfer known that do not involve mitochondria or plastids, but it is not clear whether this reflects a lack of such transfer events, or poor sampling of diverse eukaryotes. One gene where this process is apparently active is glyceraldehyde-3-phosphate dehydrogenase (GAPDH), where lateral transfer has been implicated in the origin of euglenoid and kinetoplastid genes. We have characterised GAPDH genes from diplonemids, heterotrophic flagellates that are closely related to kinetoplastids and euglenoids. Two distinct classes of diplonemid GAPDH genes were found in diplonemids, however, neither class is closely related to any other euglenozoan GAPDH. One diplonemid GAPDH is related to the cytosolic gapC of eukaryotes, although not to either euglenoids or kinetoplastids, and the second is related to cyanobacterial and proteobacterial gap3. The bacterial gap3 gene in diplonemids provides one of the most well-supported examples of lateral gene transfer from a bacterium to a eukaryote characterised to date, and may indicate that diplonemids have acquired a novel biochemical capacity through lateral transfer.

Amino Acid Sequence↗

The microorganism community in an aerobic denitrification membrane bioreactor by 16S rDNA based molecular biotechnology.

In this study, the performance and microbial community in a submerged membrane bioreactor were investigated for different hydraulic retention times. The removal efficiencies of chemical oxygen demand, total phosphorus, suspended solid and heterotrophic bacteria did not differ significantly with different hydraulic retention times but the total nitrogen removal efficiency increased when the hydraulic retention time increased. Aerobic denitrification occurred during the membrane bioreactor process at hydraulic retention time of 20 hours and Thiosphera pantotropha was identified as the aerobic denitrifier by the clone-denaturing gradient gel electrophoresis method. Another denitrifying Fe+2 oxidizing bacteria was also identified at hydraulic retention time of 20 hours. In contrast, no denitrifying bacteria were identified at hydraulic retention time of 5 hours.

Bacteria, Aerobic↗

Source bioaerosol concentration and rRNA gene-based identification of microorganisms aerosolized at a flood irrigation wastewater reuse site.

Reuse of partially treated domestic wastewater for agricultural irrigation is a growing practice in arid regions throughout the world. A field sampling campaign to determine bioaerosol concentration, culturability, and identity at various wind speeds was conducted at a flooded wastewater irrigation site in Mexicali, Baja California, Mexico. Direct fluorescent microscopy measurements for total microorganisms, culture-based assays for heterotrophs and gram-negative enteric bacteria, and small-subunit rRNA gene-based cloning were used for microbial characterizations of aerosols and effluent wastewater samples. Bioaerosol results were divided into two wind speed regimens: (i) below 1.9 m/s, average speed 0.5 m/s, and (ii) above 1.9 m/s, average speed 4.5 m/s. Average air-borne concentration of total microorganisms, culturable heterotrophs, and gram-negative enteric bacteria were, respectively, 1.1, 4.2, and 6.2 orders of magnitude greater during the high-wind-speed regimen. Small-subunit rRNA gene clone libraries processed from samples from air and the irrigation effluent wastewater during a high-wind sampling event indicate that the majority of air clone sequences were more than 98% similar to clone sequences retrieved from the effluent wastewater sample. Overall results indicate that wind is a potential aerosolization mechanism of viable wastewater microorganisms at flood irrigation sites.

Aerosols↗

Quantum thermodynamics approach to phosphorylation and heterotrophic growth yields.

A model of cell growth is presented which is based on the double postulates of quantized loss of energy during phosphorylation and reversible biosynthesis of cell structure. An immediate consequence of the postulates is the identical value for the energy efficiency of the phosphorylation and for that of the whole growth process. Another consequence is the relationship between the energy level of the biomass and the phosphorylation potential as embodied in the equation: EO = gamma'M X EATP, where EO is the heat of transfer of a pair of electrons to oxygen, EATP, the molar heat of hydrolysis of ATP, and gamma'M, the degree of reduction of the biomass, gamma M being constant and equal to 5. The model predicts five levels of growth yields corresponding to five permissible values for the P/O ratio (r = 0, 1, 2, 3, and 4). Any growth process would be characterized by a set of two integers N and lambda; N is the maximal P/O ratio prescribed by the energy content of the substrate as compared with that of the biomass, and lambda the number of further downward quantum jumps of the P/O ratio resulting from the adversity of the growth condition (N - lambda = r). Under full aerobiosis, one has 0 less than or equal to lambda less than or equal to N less than or equal to 3. When growth is limited only by the energy content of the substrate (lambda = 0), the time-independent dispersion of N, owing to substrate-level phosphorylations and (or) dephosphorylations, leads to effective values which are higher than the nominal ones for the yield per mole of oxygen and the heat of transfer of a pair of electrons. Under adverse conditions (lambda greater than 0), the apparent variations of the yields and the P/O ratio in function of the growth rate are shown to be an effect of the random dispersion of lambda and of the existence of a maximal rate of substrate consumption. Statistical evidence for the macroscopic quantum effect in heterotrophic growth is presented.

Adenosine Triphosphate↗

Feasibility of using ultrasonic irradiation to recover active biomass from waste activated sludge.

Under typical operating conditions, the microbial fraction of activated sludge flocs is approximately 40% by weight. The objective of this research is to evaluate the feasibility of using ultrasonic irradiation to disrupt activated sludge flocs allowing for the subsequent separation of active and inactive fractions. If separation of floc components is possible, then methods may be incorporated into wastewater treatment plant operations whereby only the inactive fraction of floc is wasted (i.e., of waste activated sludge, WAS), which in turn could increase the overall effective biological solids retention time, leading to increased process robustness with no net increase in reactor size. The results indicate that ultrasonic irradiation of WAS at 800 Wl(-1) followed by 30 min of settling can produce a supernatant with heterotrophic specific oxygen uptake rates (SOURs) of over two times the SOUR measured in the bulk mixed liquor. Under these conditions 26% of the initial heterotrophic activity was recovered within only 11% of the initial volatile mass. Similarly, autotrophic analysis revealed that nitrifying organisms, while sensitive to the effects of ultrasonic irradiation, can be separated from the activated sludge floc and recovered. An irradiation density of 200 Wl(-1) with an exposure time between 1 and 2 min produced a supernatant with a specific ammonia removal rate of over two times the initial mixed liquor rate.

Bacteria↗

Impacts of large-scale climatic disturbances on the terrestrial carbon cycle.

BACKGROUND: The amount of carbon dioxide in the atmosphere steadily increases as a consequence of anthropogenic emissions but with large interannual variability caused by the terrestrial biosphere. These variations in the CO2 growth rate are caused by large-scale climate anomalies but the relative contributions of vegetation growth and soil decomposition is uncertain. We use a biogeochemical model of the terrestrial biosphere to differentiate the effects of temperature and precipitation on net primary production (NPP) and heterotrophic respiration (Rh) during the two largest anomalies in atmospheric CO2 increase during the last 25 years. One of these, the smallest atmospheric year-to-year increase (largest land carbon uptake) in that period, was caused by global cooling in 1992/93 after the Pinatubo volcanic eruption. The other, the largest atmospheric increase on record (largest land carbon release), was caused by the strong El Niño event of 1997/98. RESULTS: We find that the LPJ model correctly simulates the magnitude of terrestrial modulation of atmospheric carbon anomalies for these two extreme disturbances. The response of soil respiration to changes in temperature and precipitation explains most of the modelled anomalous CO2 flux. CONCLUSION: Observed and modelled NEE anomalies are in good agreement, therefore we suggest that the temporal variability of heterotrophic respiration produced by our model is reasonably realistic. We therefore conclude that during the last 25 years the two largest disturbances of the global carbon cycle were strongly controlled by soil processes rather then the response of vegetation to these large-scale climatic events.

Journal Article↗

[Regulation of metabolism in microorganisms with CO2 fixation].

The paper deals with a review of the data available in literature on the regulation of the metabolic processes in microorganisms with CO2 fixation. The main function of CO2 is traced as exemplified by different types of microorganism nutrition. The carboxylation reactions in their main forms are observed in all heterotrophic microorganisms. The experimental data relative to the heterotrophic fixation of CO2 in yeast assimilating oil carbohydrates are presented as an example. The degree of CO2 fixation under these conditions may reach 30%. Carbon of CO2 is utilized for the biomass biosynthesis.

Carbohydrate Metabolism↗

Plastids and protein targeting.

Plastids with two bounding membranes--as exemplified by red algae, green algae, plants, and glaucophytes--derive from primary endosymbiosis; a process involving engulfment and retention of a cyanobacterium by a phagotrophic eukaryote. Plastids with more than two bounding membranes (such as those of euglenoids, dinoflagellates, heterokonts, haptopytes, apicomplexa, cryptomonads, and chlorarachniophytes) probably arose by secondary endosymbiosis, in which a eukaryotic alga (itself the product of primary endosymbiosis) was engulfed and retained by a phagotroph. Secondary endosymbiosis transfers photosynthetic capacity into heterotrophic lineages, has apparently occurred numerous times, and has created several major eukaryotic lineages comprising upwards of 42,600 species. Plastids acquired by secondary endosymbiosis are sometimes referred to as "second-hand." Establishment of secondary endosymbioses has involved transfer of genes from the endosymbiont nucleus to the secondary host nucleus. Limited gene transfer could initially have served to stabilise the endosymbioses, but it is clear that the transfer process has been extensive, leading in many cases to the complete disappearance of the endosymbiont nucleus. One consequence of these gene transfers is that gene products required in the plastid must be targeted into the organelle across multiple membranes: at least three for stromal proteins in euglenoids and dinoflagellates, and across five membranes in the case of thylakoid lumen proteins in plastids with four bounding membranes. Evolution of such targeting mechanisms was obviously a key step in the successful establishment of each different secondary endosymbiosis. Analysis of targeted proteins in the various organisms now suggests that a similar system is used by each group. However, rather than interpreting this similarity as evidence of an homologous origin, I believe that targeting has evolved convergently by combining and recycling existing protein trafficking mechanisms already existing in the endosymbiont and host. Indeed, by analyzing the multiple motifs in targeting sequences of some genes it is possible to infer that they originated in the plastid genome, transferred from there into the primary host nucleus, and subsequently moved into the secondary host nucleus. Thus, each step of the targeting process in "second-hand" plastids recapitulates the gene's previous intracellular transfers.

Animals↗