The carbohydrates of Phaeocystis and their degradation in the microbial food web

The ubiquity and high productivity associated with blooms of colonial Phaeocystis makes it an important contributor to the global carbon cycle. During blooms organic matter that is rich in carbohydrates is produced. We distinguish five different pools of carbohydrates produced by Phaeocystis. Like all plants and algal cells, both solitary and colonial cells produce (1) structural carbohydrates, (hetero) polysaccharides that are mainly part of the cell wall, (2) mono- and oligosaccharides, which are present as intermediates in the synthesis and catabolism of cell components, and (3) intracellular storage glucan. Colonial cells of Phaeocystis excrete (4) mucopolysaccharides, heteropolysaccharides that are the main constituent of the mucous colony matrix and (5) dissolved organic matter (DOM) rich in carbohydrates, which is mainly excreted by colonial cells. In this review the characteristics of these pools are discussed and quantitative data are summarized. During the exponential growth phase, the ratio of carbohydrate-carbon (C) to particulate organic carbon (POC) is approximately 0.1. When nutrients are limited, Phaeocystis blooms reach a stationary growth phase, during which excess energy is stored as carbohydrates. This so-called overflow metabolism increases the ratio of carbohydrate-C to POC to 0.4–0.6 during the stationary phase, leading to an increase in the C/N and C/P ratios of Phaeocystis organic matter. Overflow metabolism can be channeled towards both glucan and mucopolysaccharides. Summarizing the available data reveals that during the stationary phase of a bloom glucan contributes 0–51% to POC, whereas mucopolysaccharides contribute 5–60%. At the end of a bloom, lysis of Phaeocystis cells and deterioration of colonies leads to a massive release of DOM rich in glucan and mucopolysaccharides. Laboratory studies have revealed that this organic matter is potentially readily degradable by heterotrophic bacteria. However, observations in the field of accumulation of DOM and foam indicate that microbial degradation is hampered. The high C/N and C/P ratios of Phaeocystis organic matter may lead to nutrient limitation of microbial degradation, thereby prolonging degradation times. Over time polysaccharides tend to self-assemble into hydrogels. This may have a profound effect on carbon cycling, since hydrogels provide a vehicle to move DOM up the size spectrum to sizes subject to sedimentation. In addition, it changes the physical nature and microscale structure of the organic matter encountered by bacteria which may affect the degradation potential of the Phaeocystis organic matter.     

Limited sinking of Phaeocystis during a 12 days sediment trap study

There is a controversy discussion about the contribution of the genus Phaeocystis to the vertical carbon export with evidence for and against sedimentation of Phaeocystis. So far, the presence of Phaeocystis in sinking matter was investigated with methods depending on morphological features (microscopy) and fast degradable substances (biochemical analyses). In this study, we determine the occurrence and abundance of Phaeocystis antarctica in short‐term sediment traps and the overlying water column during a 12‐day time period in the Atlantic sector of the Southern Ocean with 454‐pyrosequencing and microscopy counting. In the sediment trap samples, we only found few sequences belonging to Phaeocystis, which was not reflecting the situation in the water column above. The cell counts showed the same results. We conclude that Phaeocystis cells are not generally transported downwards by active sinking or other sinking processes.     

Effects of iron concentration on pigment composition in Phaeocystis antarctica grown at low irradiance

Interpretation of photosynthetic pigment data using iterative programs such as CHEMTAX are widely used to examine algal community structure in the surface ocean. The accuracy of such programs relies on understanding the effects of environmental parameters on the pigment composition of taxonomically diverse algal groups. Phaeocystis antarctica is an important contributor to total autotrophic production and the biogeochemical cycling of carbon and sulfur in the Southern Ocean. Here we report the results of a laboratory culture experiment in which we examined the effects of ambient dissolved iron concentration on the pigment composition of colonial P. antarctica, using a new P. antarctica strain isolated from the southern Ross Sea in December 2003. Low-iron (<0.2 nM dissolved Fe) filtered Ross Sea seawater was used to prepare the growth media, thus allowing sub-nanomolar iron additions without the use of EDTA to control dissolved iron concentrations. The experiment was conducted at relatively low irradiance (∼20 μE m⁻² s⁻¹), with P. antarctica primarily present in the colonial form—conditions that are typical of the southern Ross Sea during austral spring. Relative to the iron-limited control treatments (0.22 nM dissolved Fe), iron addition mediated a decrease in the ratio of 19′-hexanoyloxyfucoxanthin to chlorophyll a, and an increase in the ratio of fucoxanthin to chlorophyll a. Our results also suggest that the ratio of 19′-hexanoyloxyfucoxanthin to chlorophyll c₃ (Hex:Chl c₃ ratio) may be a characteristic physiological indicator for the iron-nutritional status of colonial P. antarctica, with higher Hex:Chl c₃ ratios (>3) indicative of Fe stress. We also observed that the ratio of fucoxanthin to 19′-hexanoyloxyfucoxanthin (Fuco:Hex ratio) was highly correlated (r ² = 0.82) with initial dissolved Fe concentration, with Fuco:Hex ratios <0.05 measured under iron-limited conditions (dissolved Fe <0.45 nM). Our results corroborate and extend the results of previous experimental studies, and, combined with pigment measurements from the southern Ross Sea, are consistent with the hypothesis that the interconversion of fucoxanthin and 19′-hexanoyloxyfucoxanthin by colonial P. antarctica is used as a photo-protective or light-harvesting mechanism, according to the availability of dissolved iron.     

Size‐spectrum based differential response of phytoplankton to nutrient and iron‐organic matter combinations in microcosm experiments in a Chilean Patagonian Fjord

The Patagonian fjords have been recognized as a major region of relatively high primary productivity systems during spring–summer bloom periods, where iron‐organic matter forms may be essential complexes involved in key growth processes connected to the carbon and nitrogen cycles. We used two dissolved organic matter (DOM) types, marine polysaccharide and siderophore, as a model to understand how they affect the bioavailability of Fe to phytoplankton and bacteria and to assess their ecological role in fjord systems. A 10‐day microcosm study was performed in the Comau Fjord during summer conditions (March 2012). Pico‐, nano‐, and microphytoplankton abundance, total chlorophyll‐a and bacteria abundance, and bacterial secondary production estimates were analyzed in five treatments: (i) control (no additions), (ii) only nutrients (NUT: PO₄, NO₃, Si), (iii) nutrients + Fe(II), (iv) polysaccharide (natural diatoms extracted: 1–3 beta Glucan), and (v) Hexandentate Desferroxiamine B (DFB, siderophore). Our results showed that while DFB reduced Fe bioavailability for almost all phytoplankton assemblages in the fjord, polysaccharide did not have effects on the iron bioavailability. At Nutrients + Fe and Polysaccharide treatments, chlorophyll‐a concentration abruptly increased from 0.9 to 20 mg m^?3 during the first 4–6 days of the experimental period. Remarkably, at the Nutrients + Fe treatment, the development of the bloom was accompanied by markedly high abundances of Synechococcus, picoeukaryotes, and autotrophic nanoflagellates within the first 4 days of the experiment. Our study indicated that small plankton (phytoplankton <20 μm and bacteria) were the first to respond to dissolved Nutrients + Fe compared to large sized micro‐phytoplankton cells (>20 μm). This could be at least partially attributed to biological utilization of Fe (2 to 3 nM) by <20 μm phytoplankton and bacteria through the interaction with organic ligands released by bacteria that eventually could increase solubility of the Fe dissolved fraction thus having a positive effect on the small‐sized phytoplankton community.     

Moss and Peat Leachate Degradability by Heterotrophic Bacteria: The Fate of Organic Carbon and Trace Metals

The respiration of dissolved organic matter (DOM) by aerobic heterotrophic bacterioplankton in boreal surface waters is one of the major factors that regulate CO₂ exchange of lakes and rivers with the atmosphere in arctic and subarctic zones. The DOM that originates from topsoil leaching and vegetation degradation is brought to the lakes by surface flow and is subjected to coagulation and degradation by heterotrophic bacteria, which are well-established processes in the majority of boreal aquatic settings. The behavior of colloids and organic complexes of trace metals during this process is virtually unknown. In this work, we studied the interaction of two model heterotrophic bacteria, soil Pseudomonas aureofaciens and aquatic Pseudomonas reactans, with peat and Sphagnum moss leachates from the permafrost region under controlled laboratory conditions in nutrient-free media. The moss leachate was the better substrate for bacterial survival, with P. reactans exhibiting an order of magnitude higher live cell number compared with P. aureofaciens. In eight-day experiments, we analyzed organic carbon and ～40 major and trace elements (TEs) during heterotrophic bacteria growth. The total net decrease in the concentration of dissolved organic carbon (DOC) was similar for both bacteria and ranged from 30 mg g_wet^?1 to ≤10 mg g_wet^?1 during 8 days for the moss and peat leachate, respectively. Despite significant evolutions of pH, DOC, dissolved inorganic carbon (DIC), and cell number, most major (Mg, K, and Ca) and TEs remained nearly constant (within ±30% of the control). Only Fe, Al, P, Zn, Mn, Co, and Ba and to a much lesser extent Cd, Pb, Rare Earth Elements (REEs), U, Ti, and Zr were affected (p??1 to µg L^?1 and followed the order DOC >> P >> Ba > Zn ≥ Fe ≥ Al > Mn > Cu ≥ Sr > Zr ≥ Ti > Ni ≥ Co > REEs ≥ U > Hf～Th, which reflected the abundance of the elements in the two substrates. Generally, the soil exopolysaccharide producing bacterium P. aureofaciens in the peat leachate had the greatest impact of the four combinations investigated in this study (two bacteria with two substrates). Under ongoing environmental changes in the boreal zone, the autochthonous processes of bacterioplankton activity are able to decrease the concentrations of a very limited number of TEs, including mainly Fe and several macro- (P) and micro- (Zn, Mn, and Ba) nutrients.     

Degradation of paper mill water components in laboratory tests with pure cultures of bacteria

The degradation of dissolved and colloidal substances from thermomechanical pulp (TMP) by bacteria isolated from a paper mill was studied in a laboratory slide culture system.Burkholderia cepacia strains hydrolysed triglycerides to free fatty acids, and the liberated unsaturated fatty acids were then degraded to some extent. Saturated fatty acids were not notably degraded. However, the branched anteiso-heptadecanoic fatty acid was degraded almost like the unsaturated fatty acids. About 30% of the steryl esters were degraded during 11 days, increasing the concentrations of free sterols. Approximately 25% of the dehydroabietic, and 45% of the abietic and isopimaric resin acids were degraded during 11 days. The degree of unsaturation seemed to be of greater importance for the degradation of fatty acids than the molar mass. No degradation of dissolved hemicelluloses could be observed with any of the nine bacterial strains studied. Burkholderia cepacia strains and one Bacillus coagulans strain degraded monomeric fructose and glucose in winter TMP water, but in summer TMP water, with much lower sugar concentrations, also otherBacillus strains degraded monomeric sugars.     

Does Phaeocystis spp. contribute significantly to vertical export of organic carbon?

Phaeocystis spp. cell and colony mass fluxes and their contribution to the vertical particulate organic carbon (POC) export from a wide range of stations were quantified by short-term sediment traps. The compilation of available data, ranging from polar to sub-arctic and boreal regions, revealed that Phaeocystis colonial and single cells frequently are observed in shallow sediment traps at 30–50 m depth (average of 7 ± 11% of POC export). A strong vertical export decline between 40 m and 100 m diminished the contribution of Phaeocystis spp. cell carbon to vertical export of POC to only 3 ± 2% at 100 m depth, with two exceptions (deeper mixed stations). Estimates of potential corresponding mucus contribution increased the average Phaeocystis spp. contribution to <5% of POC export. The vertical flux attenuation efficiency is higher for Phaeocystis spp. than for diatoms. The overall contribution of Phaeocystis spp. to vertical carbon export based on direct investigations of vertical organic carbon export is small.     

Bacterial consumption of total and dissolved organic carbon in the Great Barrier Reef

Heterotrophic bacteria typically take up directly dissolved organic matter due to the small molecular size, although both particulate and dissolved organic matter have labile (easily consumed) compounds. Tropical coastal waters are important ecosystems because of their high productivity. However, few studies have determined bacterial cycling (i.e. carbon uptake by bacteria and allocation for bacterial biomass and respiration) of dissolved organic carbon in coastal tropical waters, and none has determined bacterial cycling of total and dissolved organic carbon simultaneously. In this study we followed bacterial biomass and production, and organic carbon changes over short-term (12 days) dark incubations with (total organic carbon, TOC) and without particulate organic carbon additions (dissolved organic carbon, DOC). The study was performed at three sites along the middle stretch of the Great Barrier Reef (GBR) during the dry and wet seasons. Our results show that the bacterial growth efficiency is low (0.1–11.5%) compared to other coastal tropical systems, and there were no differences in the carbon cycling between organic matter sources, seasons or locations. Nonetheless, more carbon was consumed in the TOC compared to the DOC incubations, although the proportion allocated to biomass and respiration was similar. This suggests that having more bioavailable substrate in the particulate form did not benefit bacteria. Overall, our study indicates that when comparing the obtained respiration rates with previously measured primary production rates, the GBR is a heterotrophic system. More detailed studies are required to fully explore the mechanisms used by bacteria to cycle TOC and DOC in tropical coastal waters.

     

Effects of zooplankton carcasses degradation on freshwater bacterial community composition and implications for carbon cycling

Non-predatory mortality of zooplankton provides an abundant, yet, little studied source of high quality labile organic matter (LOM) in aquatic ecosystems. Using laboratory microcosms, we followed the decomposition of organic carbon of fresh ¹³C-labelled Daphnia carcasses by natural bacterioplankton. The experimental setup comprised blank microcosms, that is, artificial lake water without any organic matter additions (B), and microcosms either amended with natural humic matter (H), fresh Daphnia carcasses (D) or both, that is, humic matter and Daphnia carcasses (HD). Most of the carcass carbon was consumed and respired by the bacterial community within 15 days of incubation. A shift in the bacterial community composition shaped by labile carcass carbon and by humic matter was observed. Nevertheless, we did not observe a quantitative change in humic matter degradation by heterotrophic bacteria in the presence of LOM derived from carcasses. However, carcasses were the main factor driving the bacterial community composition suggesting that the presence of large quantities of dead zooplankton might affect the carbon cycling in aquatic ecosystems. Our results imply that organic matter derived from zooplankton carcasses is efficiently remineralized by a highly specific bacterial community, but does not interfere with the bacterial turnover of more refractory humic matter.     

Evidence for high iron requirements of colonial Phaeocystis antarctica at low irradiance

We have carried out field and laboratory experiments to examine the iron requirements of colonial Phaeocystis antarctica in the Ross Sea. In December 2003, we performed an iron/light-manipulation bioassay experiment in the Ross Sea polynya, using an algal assemblage dominated by colonial Phaeocystis antarctica, collected from surface waters with an ambient dissolved Fe concentration of ∼0.4 nM. Results from this experiment suggest that P. antarctica growth rates were enhanced at high irradiance (∼50% of incident surface irradiance) but were unaffected by iron addition, and that elevated irradiance mediated a significant decrease in cellular chlorophyll a content. We also conducted a laboratory iron dose–response bioassay experiment using a unialgal, non-axenic strain of colonial P. antarctica and low-iron (<0.2 nM) filtered seawater, both collected from the Ross Sea polynya in December 2003. By using rigorous trace-metal clean techniques, we performed this dose–response iron-addition experiment at ∼0°C without using organic chelating reagents to control dissolved iron levels. At the relatively low irradiance of this experiment (∼20 μE m⁻² s⁻¹), estimated nitrate-specific growth rate as a function of dissolved iron concentration can be described by a Monod relationship, yielding a half-saturation constant with respect to growth of 0.45 nM dissolved iron. This value is relatively high compared to reported estimates for other Antarctic phytoplankton. Our results suggest that seasonal changes in the availability of both iron and light play critical roles in limiting the growth and biomass of colonial Phaeocystis antarctica in the Ross Sea polynya.     

Methods used to reveal genetic diversity in the colony-forming prymnesiophytes Phaeocystis antarctica, P. globosa and P. pouchetii—preliminary results

Previous work on the genetic diversity of Phaeocystis used ribosomal DNA and internal transcribed spacer (ITS) sequence analyses to show that there is substantial inter- and intraspecific variation within the genus. First attempts to trace the biogeographical history of strains in Antarctic coastal waters were based on a comparison of ITS sequences. To gain deeper insights into the population structure and bloom dynamics of this microalga it is necessary to quantify the genetic diversity within populations of P. antarctica from different locations (i.e., each of the three major gyres in the Antarctic continental waters) and to calculate the gene flow between them. Here we describe methods to quantify genetic diversity and our preliminary results for P. antarctica in comparison to two other colonial species: P. globosa and P. pouchetii. For this study of genetic diversity, two fingerprinting techniques were used. First, amplified fragment-length polymorphisms (AFLPs) were established as a pre-screening tool to assess clone diversity and to select divergent clones prior to physiological investigations. Second, the more-powerful microsatellite markers were established to assess population structure and biogeography more accurately. Results show differences in the AFLP patterns between isolates of P. antarctica from different regions, and that a wide variety of microsatellite motifs could be obtained from the three Phaeocystis species.     

Littoral-pelagial interchange and the decomposition of dissolved organic matter in a polyhumic lake

The small, polyhumic lake, Mekkojärvi (southern Finland), is bordered by a moss vegetation zone (Warnstorfia and Sphagnum species) which provides a habitat-rich and productive environment for many planktonic and periphytic animals. Impacts of moss on the metabolism of bacterioplankton, phytoplankton and zooplankton in polyhumic water were investigated in laboratory throughflow systems. Growing Warnstorfia (together with epiphytic algae and bacteria) suppressed the production of planktonic algae but had no clear effect on leucine uptake, and hence bacterial production, or on the decomposition of humic substances. Phenol uptake and mineralization rates, however, were lower in the littoral water than in the pelagial water. Excretion of organic carbon by Warnstorfia algae or Daphnia longispina (the predominant crustacean in the pelagial water) provided only a minor contribution to bacterial production; therefore, a major contribution had to be from humic substances. A bacterial production efficiency of 31–38% could account for the microbial respiration in the water. The results indicated that bacterial, or detrital matter (originating largely from the littoral zone), could not obviate the need for algal food, and that a great deal of particulate matter in the water was poor or useless food for Daphnia. In all, the bulk of dissolved organic matter in Lake Mekkojärvi was biochemically highly recalcitrant. Our results indicate that humic substances (from watershed or littoral area) which, through bacterial degradation, enter the planktonic food web of the lake are mainly lost through respiration by microorganisms.     

Bacterial Contribution to Dissolved Organic Matter in Eutrophic Lake Kasumigaura,Japan

Incubation experiments using filtered waters from Lake Kasumigaura were conducted to examine bacterial contribution to a dissolved organic carbon (DOC) pool. Bacterial abundance, bacterial production, concentrations of DOC, total dissolved amino acids (TDAA), and total dissolved neutral sugars (TDNS) were monitored during the experiments. Bacterial production during the first few days was very high (20 to 35 μg C liter⁻¹ day⁻¹), accounting for 40 to 70% of primary production. The total bacterial production accounted for 34 to 55% of the DOC loss during the experiment, indicating high bacterial activities in Lake Kasumigaura. The DOC degradation was only 12 to 15%, whereas the degradation of TDAA and TDNS ranged from 30 to 50%, suggesting the preferential usage of TDAA and TDNS. The contribution of bacterially derived carbon to a DOC pool in Lake Kasumigaura was estimated using d-amino acids as bacterial biomarkers and accounted for 30 to 50% of the lake DOC. These values were much higher than those estimated for the open ocean (20 to 30%). The ratio of bacterially derived carbon to bulk carbon increased slightly with time, suggesting that the bacterially derived carbon is more resistant to microbial degradation than bulk carbon. This is the first study to estimate the bacterial contribution to a DOC pool in freshwater environments. These results indicate that bacteria play even more important roles in carbon cycles in freshwater environments than in open oceans and also suggests that recent increases in recalcitrant DOC in various lakes could be attributed to bacterially derived carbon. The potential differences in bacterial contributions to dissolved organic matter (DOM) between freshwater and marine environments are discussed.     

Compositions isotopiques naturelles des bactéries hétérotrophes et détermination de l'origine du carbone organique dissous biodisponible

Several studies of salt marsh systems have attempted to quantify the flow of organic matter between the land and coastal waters. However, the techniques used could not identify sources of dissolved organic carbon (DOC) rapidly assimilated by heterothrophic bacteria. Recently, the assay of carbon isotope ratios has allowed characterization of the different sources of organic matter in salt marshes. In this study, we wanted to find out if the natural isotopic composition assayed in heterotrophic bacteria distinguished the origin of bioavailable DOC. We determined the δ¹³C values for 1) three bacterial strains and their nucleic acids cultured on glucose and tryptose substrates, respectively, and 2) naturally occurring bacteria recovered from seawater in which salt marsh vegetation had been immersed. First, we showed that the isotopic fractionation was the same for the three bacterial strains cultured on the same synthetic substrate, but could vary depending on the nature of DOC. There was no significant difference between the δ¹³ C values of bacteria and their nucleic acids. Second, natural bacteria were grown in a medium enriched in DOC from halophytic plants. The δ¹³C values of this community were close to those of dissolved organic carbon from plant leachates. The comparison between the isotopic ratios of natural bacteria in Vibrio alginolyticus showed that the heterogeneity of the bacterial community averaged the isotopic fractionation from the preferential assimilation of organic compounds in the medium by each bacterial strain. The δ¹³ C values recorded for the bacterial community in the field and their nucleic acids made it possible to identify the source of organic matter readily accessible to microorganisms in a coastal ecosystem.     

Phaeocystis and its interaction with viruses

Over the years, viruses have been shown to be mortality agents for a wide range of phytoplankton species, including species within the genus Phaeocystis (Prymnesiophyceae). With its polymorphic life cycle, its worldwide distribution, and the capacity of several of the Phaeocystis species to form dense blooms, this genus is a key player for our understanding of biogeochemical cycling of elements. This paper provides an overview of what is know to date about the ecological role of viruses in regulating Phaeocystis population dynamics. It explores which variables affect the algal host–virus interactions, and examines the impact of virally induced cell lysis of Phaeocystis on the function and structure of the pelagic food web as well as on the flow of organic carbon and nutrients.     

The influence of dietary iron on zinc in rat

The purpose of this study was to clarify the influence of iron on zinc status. The animals were divided into four groups, consisting of five rats in each group. The control group was fed on basal diet with adequate levels of zinc and iron, whereas the experimental group was fed diets containing different levels of iron ad libitum for 15 d. Low levels of iron (LFe) significantly increased the zinc absorption percentage but there was a decrease in high (HFe) and very high iron (VHFe) level groups (p<0.001). The retention percentage changes were found to be parallel to the changes in the absorption percentage curve. It was found that zinc (per total dry tissue) and Zn-65 (per total tissue) increased in the rats fed the LFe, whereas in general they decreased in the rats fed the HFe and VHFe diets. Significant changes were found in the duodenum and liver. Zn-65 (per g wet tissue) significantly increased in the brain and liver in the LFe group, but there was a decrease in the duodenum, ileum, kidney, liver, and brain in the HFe and VHFe groups. Changes in the level of zinc (per g dried tissue) were found to be parallel to the changes in Zn-65 in all the groups. The dietary proportions of iron appear to influence zinc metabolism at the intestinal and cellular transport levels over a given period of time.     

Biodegradation of naphthalene-2-sulfonic acid present in tannery wastewater by bacterial isolates <Emphasis Type="Italic">Arthrobacter</Emphasis> sp. 2AC and <Emphasis Type="Italic">Comamonas</Emphasis> sp. 4BC

Two bacterial strains, 2AC and 4BC, both capable of utilizing naphthalene-2-sulfonic acid (2-NSA) as a sole source of carbon, were isolated from activated sludges previously exposed to tannery wastewater. Enrichments were carried out in mineral salt medium (MSM) with 2-NSA as the sole carbon source. 16S rDNA sequencing analysis indicated that 2AC is an Arthrobacter sp. and 4BC is a Comamonas sp. Within 33 h, both isolates degraded 100% of 2-NSA in MSM and also 2-NSA in non-sterile tannery wastewater. The yield coefficient was 0.33 g biomass dry weight per gram of 2-NSA. A conceptual model, which describes the aerobic transformation of organic matter, was used for interpreting the biodegradation kinetics of 2-NSA. The half-lives for 2-NSA, at initial concentrations of 100 and 500 mg/l in MSM, ranged from 20 h (2AC) to 26 h (4BC) with lag-phases of 8 h (2AC) and 12 h (4BC). The carbon balance indicates that 75–90% of the initial TOC (total organic carbon) was mineralized, 5–20% remained as DOC (dissolved organic carbon) and 3–10% was biomass carbon. The principal metabolite of 2-NSA biodegradation (in both MSM and tannery wastewater) produced by Comamonas sp. 4BC had a MW of 174 and accounted for the residual DOC (7.0–19.0% of the initial TOC and 66% of the remaining TOC). Three to ten percent of the initial TOC (33% of the remaining TOC) was associated with biomass. The metabolite was not detected when Arthrobacter sp. 2AC was used, and a lower residual DOC and biomass carbon were recorded. This suggests that the two strains may use different catabolic pathways for 2-NSA degradation. The rapid biodegradation of 2-NSA (100 mg/l) added to non-sterile tannery wastewater (total 2-NSA, 105 mg/l) when inoculated with eitherArthrobacter 2AC or Comamonas 4BC showed that both strains were able to compete with the indigenous microorganisms and degrade 2-NSA even in the presence of alternate carbon sources (DOC in tannery wastewater = 91 mg/l). The results provide information useful for the rational design of bioreactors for tannery wastewater treatment.     

Microbial Communities Involved in Fe Reduction and Mobility During Soil Organic Matter (SOM) Mineralization in Two Contrasted Paddy Soils

Lowland rice fields of West Africa (Ivory Coast) and South Asia (Thailand) are affected by ferrous toxicity or salinity, respectively, and their soil waters contain large amounts of ferrous iron, depending on reducing irrigation condition and suggesting occurrence of bacterial reducing processes. To determine the involvement, dynamic and activities of bacterial communities in Fe(III) reduction and mobilization during anaerobic degradation and mineralization of soil organic matter (SOM), different experiments and analyses have been performed. Results demonstrated that the utilization of SOM as sole carbon, nutrient and energy sources favored the presence of large bacterial communities: facultative anaerobic and anaerobic bacteria, Fe(III)-reducing bacteria (FeRB) (fermentative and Fe respiring), sulfate reducing bacteria (SRB) which are involved in carbon, nitrogen, iron and sulfur cycling. The larger functional diversity is observed in the Ivory Coast paddy soils containing larger amounts of organic matter and sulfur compounds. These communities contained complementary populations (chemoorganotrophic, chemolitotrophic, aerobic, facultative anaerobic and anaerobic) that can be active at different steps of iron solubilization with simultaneous organic matter mineralization. Our results indicate that the pH controlled by bacterial activity, the nature much more than the content of organic matter, and consequently the structure and activity of bacterial communities influence significantly the availability and dynamic of iron in paddy fields which affect the soil quality.     

Toluene biodegradation by <Emphasis Type="Italic">Pseudomonas putida</Emphasis> F1: targeting culture stability in long-term operation

The stability of Pseudomonas putida F1, a strain harbouring the genes responsible for toluene degradation in the chromosome was evaluated in a bioscrubber under high toluene loadings and nitrogen limiting conditions at two dilution rates (0.11 and 0.27 h⁻¹). Each experiment was run for 30 days, period long enough for microbial instability to occur considering previously reported studies carried out with bacterial strains encoding the catabolic genes in the TOL plasmid. At all tested conditions, P. putida F1 exhibited stable performance as shown by the constant values of the specific toluene degradation yield, CO₂ produced versus toluene degraded yield, and biomass concentration within each steady state. Benzyl alcohol, a curing agent causing TOL plasmid deletion in Pseudomonas strains, was present in the cultivation medium as a result of the monooxygenation of toluene by the diooxygenase system of P. putida F1. However, no mutant population growing at the expense of the extracellular excreted carbon or lysis products was established in the chemostat as confirmed by the constant dissolved total organic carbon (TOC) concentration and fraction of toluene degrading cells (approx. 100%). In addition, batch experiments conducted with both lysis substrate and toluene simultaneously confirmed that P. putida F1 preferentially consumed toluene rather than extracellular excreted carbon.     

Bacterial Growth on Allochthonous Carbon in Humic and Nutrient-enriched Lakes: Results from Whole-Lake 13C Addition Experiments

Organic carbon (C) in lakes originates from two distinct sources—primary production from within the lake itself (autochthonous supply) and importation of organic matter from the terrestrial watershed (allochthonous supply). By manipulating the ¹³C of dissolved inorganic C, thereby labeling within-lake primary production, we examined the relative importance of autochthonous and allochthonous C in supporting bacterial production. For 35 days, NaH¹³CO₃ was added daily to two small, forested lakes. One of the lakes (Peter) was fertilized so that primary production exceeded total respiration in the epilimnion. The other lake (Tuesday), in contrast, was low in productivity and had high levels of colored dissolved organic C (DOC). To obtain bacterial C isotopes, bacteria were regrown in situ in particle-free lake water in dialysis tubes. The contribution of allochthonous C to bacterial biomass was calculated by applying a two-member mixing model. In the absence of a direct measurement, the isotopic signature of the autochthonous end-member was estimated indirectly by three different approaches. Although there was excess primary production in Peter Lake, bacterial biomass consisted of 43–46% allochthonous C. In Tuesday Lake more than 75% of bacterial growth was supported by allochthonous C. Although bacteria used autochthonous C preferentially over allochthonous C, DOC from the watershed contributed significantly to bacterial production. In combination with results from similar experiments in different lakes, our findings suggest that the contribution of allochthonous C to bacterial production can be predicted from ratios of chromophoric dissolved organic matter (a surrogate for allochthonous supply) and chlorophyll a (a surrogate for autochthonous supply).