Continuous Culture Experiment Research Articles

Uncovering the role of oxygen on organic carbon cycling: insights from a continuous culture study with a facultative anaerobic bacterioplankton species (Shewanella baltica)

Deoxygenation is tied to organic carbon (Corg) supply and utilization in marine systems. Under oxygen-depletion, bacteria maintain Corg respiration using alternative electron acceptors such as nitrate. Since anaerobic respiration’s energy yield is lower, Corg remineralization may be reduced and its residence time increased. We investigated the influence of oxygen and alternative electron acceptors’ availability on Corg cycling by heterotrophic bacteria during a continuous culture experiment with Shewanella baltica, a facultative anaerobic γ-Proteobacteria in the Baltic Sea. We tested six different oxygen levels, from suboxic (<5 µmol L-1) to fully oxic conditions, using a brackish (salinity=14 g L-1) media supplied with high (HighN) or low (LowN) inorganic nitrogen concentrations relative to glucose as labile Corg source. Our results show that suboxia limited DOC (glucose) uptake and cell growth only under LowN, while higher availability of alternative electron acceptors seemingly compensated oxygen limitation under HighN. N-loss was observed under suboxia in both nitrogen treatments. Under HighN, N-loss was highest and a C:N loss ratio of ~2.0 indicated that Corg was remineralized via denitrification. Under LowN, the C:N loss ratio under suboxia was higher (~5.5), suggesting the dominance of other anaerobic respiration pathways, such as dissimilatory nitrate reduction to ammonium (DNRA). Bacterial growth efficiency was independent of oxygen concentration but higher under LowN (34 ± 3.0%) than HighN (26 ± 1.6%). Oxygen concentration also affected dissolved organic matter (DOM) cycling. Under oxic conditions, the release of dissolved combined carbohydrates was enhanced, and the amino acid-based degradation index (DI) pointed to more diagenetically altered DOM. Our results suggest bacterial Corg uptake in low-oxygen systems dominated by S. baltica can be limited by oxygen but compensated by high nitrate availability. Hence, suboxia diminishes Corg remineralisation only when alternative electron acceptors are lacking. Under high nitrate:Corg supply, denitrification leads to a higher N:C loss ratio, potentially counteracting eutrophication in the long run. Low nitrate:Corg supply may favour other anaerobic respiration pathways like DNRA, which sustains labile nitrogen in the system, potentially intensifying the cycle of eutrophication. Going forward, it will be crucial to establish the validity of our findings for S. baltica in natural systems with diverse organic substrates and microbial consortia.

Effect of increased pCO2 and temperature on the phytoplankton community in the coastal of Yellow Sea

In order to study the dynamics of marine phytoplankton communities in response to anticipated in temperature and CO2, a shipboard continuous culture experiment (Ecostat) was conducted. The experiment involved simulations under current atmospheric CO2 concentrations (400 ppm) and projected year-2100 CO2 levels (1000 ppm), as well as varying temperature under present (22 °C) versus increased temperature (26 °C) in the Yellow Sea during the summer of 2020. The results showed that both the increased pCO2 and temperature had significant effects on microphytoplankton and picophytoplankton, with the warming effect proving to be more significant. The different responses of various species to acidification and warming and their coupling effect led to the changes in microphytoplankton and picophytoplankton community structure. Elevated temperature and greenhouse treatments promoted the growth of dominant diatoms and Synechococcus, such as Guinardia flaccida and Pseudo-nitzschia delicatissima. This phenomenons widened the ecological niche, and the changes in the growth patterns of dominant species consequently influenced the content of cellular elements. Mantel's analysis further demonstrated that both warming and greenhouse promoted the growth of diatoms and Synechococcus. Projections of marine phytoplankton community trends by the end of the century based on Growth Rate Ratio (GRR), indicated that not only would species with GRR < 1 decrease, but also numerous species with growth rates >1 at elevated pCO2 levels would be ousted from competition. This experiment demonstrates the need to investigate whether extended exposure to increased pCO2 and temperature over more extended time scales would similarly induce shifts in the biological and biogeochemical dynamics of the Yellow Sea.

Evaluation of dietary addition of 2 essential oils from Achillea moschata, or their components (bornyl acetate, camphor, and eucalyptol) on invitro ruminal fermentation and microbial community composition.

This study investigated the effects of 2 Achillea moschata essential oils extracted from plants collected in 2 different valleys of the Italian Alps and 3 pure compounds of oils — bornyl acetate (BOR), camphor (CAM), and eucalyptol (EUCA) — on in vitro ruminal fermentation and microbiota. An in vitro batch fermentation experiment (Exp. 1) tested the addition of all of the substances (2 essential oils and 3 compounds) in fermentation bottles (120 mL) at 48 h of incubation, whereas a subsequent in vitro continuous culture experiment (Exp. 2) evaluated the pure compounds added to the fermenters (2 L) for a longer incubation period (9 d). In both experiments, total mixed rations were incubated with the additives, and samples without additives were included as the control (CTR). Each treatment was tested in duplicate and was repeated in 3 and 2 fermentation runs in Exp. 1 and 2, respectively. Gas production (GP) in Exp. 1 was similar for all of the treatments, and short chain volatile fatty acid (SCFA) production was similar in both experiments except for a decrease of SCFA produced (P = 0.029) due to EUCA addition in Exp. 2. Compared to CTR, BOR and CAM reduced the valerate proportion (P = 0.04) in Exp. 1, and increased (P < 0.01) the acetate proportion in Exp. 2. All treatments increased (P < 0.01) total protozoa counts (+36.7% and +48.4% compared to CTR on average for Exp. 1 and 2, respectively). In Exp. 1, all of the treatments lowered the Bacteroidetes and Firmicutes and increased the Proteobacteria relative abundances (P < 0.05), whereas in Exp. 2, the EUCA addition increased (P = 0.012) the Ruminococcus. In Exp. 1, methane (CH4) as a proportion of the GP was lowered (P = 0.004) by the addition of CAM and EUCA compared to CTR, whereas in Exp. 2, EUCA reduced the amount of stoichiometrically calculated CH4 compared to CTR. Overall, essential oils extracted from A. moschata and the pure compounds did not depress in vitro rumen fermentation, except for EUCA in Exp. 2. In both experiments, an increase of the protozoal population occurred for all the additives.

Effects of Nano-magnesium Hydroxide on the Forms of Cadmium in Different Types of Soil

The effects of nano-magnesium hydroxide and common magnesium hydroxide (100, 200, and 300 mg·kg-1) on the forms of cadmium in different types of cadmium contaminated soils (1, 5, 10, and 15 mg·kg-1) were studied under 28 days of continuous culture experiment. In the neutral soil, during the 28 days of culture, soil exchange Cd (EX-Cd) form distribution ratio (FDC) decreased at first and then increased with the culture time increasing under treatment of 1, 5, 10, and 15 mg·kg-1 Cd. The minima of soil EX-Cd FDC were found on the 14th day under 1 mg·kg-1 Cd and 5 mg·kg-1 Cd treatments, whereas the minima of soil EX-Cd FDC were observed on the 4th day under 10 mg·kg-1 Cd and 15 mg·kg-1 Cd treatments. The FDC of soil carbonate bound Cd (CAB-Cd), iron manganese oxidized Cd (FeMn-Cd), and organic bound Cd (OM-Cd) increased at first, then decreased, and finally, became stable, and the maxima of soil CAB-Cd, FeMn-Cd, and OM-Cd FDC were found on the 4th day, whereas the minima of soil CAB-Cd, FeMn-Cd, and OM-Cd FDC were observed on the 14th day. Soil residual Cd (RES-Cd) FDC increased gradually and then tended to becomes stable during the 28 days of culture. The soil EX-Cd FDC was 66.7%-81.8% at 1, 5, 10, and 15 mg·kg-1 Cd treatments, which was the main form of the soil. The FDC of soil Cd forms was in the order of EX-Cd > CAB-Cd > RES-Cd > FeMn-Cd > OM-Cd. Soil EX-Cd FDC reached the lowest value on the 14th Day. Soil EX-Cd FDC was reduced by nano-magnesium hydroxide and common magnesium hydroxide, and it decreased with the increase of the amount of magnesium hydroxide. During 0-28 days of culture, the soil EX-Cd FDC decreased by 11.4%-67.7%, 7.8%-37.2%, 7.7%-36.4%, 5.0%-28.8% (nano-magnesium hydroxide) and 0.5%-49.5%, 0.6%-15.0%, 1.0%-18.1%, 0.7%-14.6% (ordinary magnesium hydroxide) at 1, 5, 10, and 15 mg·kg-1 Cd treatments, respectively. The EX-Cd content of alkaline soil reached the lowest value on the 7th day of culture, and the EX-Cd content of acidic soil reached the lowest value on the 21st day under 1, 5, and 10 mg·kg-1 Cd treatments. The content of EX-Cd in neutral, acidic, and alkaline soils decreased with the increase of magnesium hydroxide content, and the content of EX-Cd in soil decreased with the increase of magnesium hydroxide amount. At the same amount, the effect of passivating soil EX-Cd under nanometer magnesium hydroxide treatment was superior to ordinary magnesium hydroxide treatment.

Antibiotic disturbance affects aquatic microbial community composition and food web interactions but not community resilience.

Notwithstanding the fundamental role that environmental microbes play for ecosystem functioning, data on how microbes react to disturbances are still scarce, and most factors that confer stability to microbial communities are unknown. In this context, antibiotic discharge into the environment is considered a worldwide threat for ecosystems with potential risks to human health. We therefore tested resilience of microbial communities challenged by the presence of an antibiotic. In a continuous culture experiment, we compared the abundance, composition and diversity of microbial communities undisturbed or disturbed by the constant addiction of tetracycline in low (10µg/L) or intermediate (100µg/L) concentration (press disturbance). Further, the bacterial communities in the three treatments had to face the sudden pulse disturbance of adding an allochthonous bacterium (Escherichia coli). Tetracycline, even at low concentrations, affected microbial communities by changing their phylogenetic composition and causing cell aggregation. This, however, did not coincide with a reduced microbial diversity, but was mainly caused by a shift in dominance of specific bacterial families. Moreover, the less disturbed community (10µg/L tetracycline) was sometimes more similar to the control and sometimes more similar to heavily disturbed community (100µg/L tetracycline). All in all, we could not see a pattern where the communities disturbed with antibiotics were less resilient to a second disturbance introducing E. coli, but they seemed to be able to buffer the input of the allochthonous strain in a similar manner as the control.

유기염소계 난분해성 산업폐수의 처리를 위한 미생물제제의 개발

유기염소계 난분해성 산업폐수처리에 효과적인 미생물제제 개발을 위하여 PCP (pentachlorophenol)와 TCE (trichloroethylene) 등과 같은 유기계 염소화합물이 오염되어있는 토양 및 산업폐수로부터 PCP 분해활성이 높은 GP5, GP19와 TCE 분해활성이 높은 GA6, GA15를 분리하였다. 이들 분리균주 GP5, GP19, GA6, GA15등은 Acetobactor sp., Pseudomonas sp., Arthrobacer sp., Xanthomonas sp.과 유사한 것으로 나타나 최종적으로 Acetobacter sp. GP5, Pseudomonas sp. GP19, Arthrobacer sp. GA6, Xanthomoas sp. GA15로 명명하였다. 유기염소계 산업폐수의 처리를 위한 복합미생물제제 OC17은 PCP와 TCE를 분해하는 4개의 분리 분리균주와 방향족화합물 분해균주인 Acinetobacter sp. KN11, Neisseria sp. GN13의 배양액을 혼합하여 제조하였다. 복합미생물제제 OC17은 <TEX>$2.8{\times}10^9CFU/g$</TEX>의 균체수를 갖고 있으며, 밀도는 <TEX>$0.299g/cm^3$</TEX>, 수분함량은 26.8%를 나타내었다. 복합미생물 제제 OC17은 PCP 500 mg/l가 포함되어있는 인공폐수를 이용한 실험에서 배양 65시간에 87%의 분해효율을 나타내었고, TCE (300 uM)의 분해효율은 배양 50시간에 90%의 분해효율을 나타내었다. 복합미생물제제 OC17을 이용한 유기 염소계 산업폐수의 처리효율 시험을 위한 연속배양 실험 에서 10일간 처리 하였을 때 91%의 COD 제거효율을 나타내었다. The screening of the microorganisms degrading chlorinated organic compounds such as PCP (pentachlorophenol) and TCE (trichloroethylene) was conducted with soil and industrial wastewater contaminated with various chlorinated organic compounds. Isolates (GP5, GP19) capable of degrading PCP and isolates (GA6, GA15) capable of degrading TCE were identified as Acetobactor sp., Pseudomonas sp., Arthrobacer sp., Xanthomonas sp. and named Acetobacter sp. GP5, Pseudomonas sp. GP19, Arthrobacer sp. GA6 and Xanthomoas sp. GA15, respectively. The microbial augmentation, OC17 formulated with the mixture of bacteria including isolates (4 strains) degrading chlorinated organic compounds and isolates (Acinetobacter sp. KN11, Neisseria sp. GN13) degrading aromatic hydrocarbons. Characteristics of microbial augmentation OC-17 showed cell mass of <TEX>$2.8{\times}10^9CFU/g$</TEX>, bulk density of <TEX>$0.299g/cm^3$</TEX> and water content of 26.8%. In the experiment with an artificial wastewater containing PCP (500 mg/l), degradation efficiency of the microbial augmentation OC17 was 87% during incubation of 65 hours. The degradation efficiency of TCE (300 uM) by microbial augmentation OC17 was 90% during incubation of 50 hours. In a continuous culture experiment, analysis of the biodegradation of organic compounds by microbial augmentation OC17 in industry wastewater containing chlorinated hydrocarbons showed that the removal rate of COD was 91% during incubation of 10 days. These results indicate that it is possible to apply the microbial augmentation OC17 to industrial wastewaters containing chlorinated organic compounds.

Comparison of alfalfa, birdsfoot trefoil, and cicer milkvetch in combination with 25, 50, or 75% tall fescue in a continuous-culture system

ABSTRACT A continuous-culture experiment was performed to investigate the effects of tall fescue (TF)-to-legume ratios (TF:legume = 75:25, 50:50, or 25:75 on a DM basis) of 3 different TF-legume mixed diets [TF-alfalfa (TF+AF), TF-birdsfoot trefoil (TF+BT), or TF-cicer milkvetch (TF+CM)] on in vitro fermentation characteristics. Nine dietary treatments were tested in a 3 (TF-legume ratio) × 3 (TF-legume mixed diet) split-plot design. Dietary treatments did not affect the concentrations of total VFA, acetate, and butyrate, whereas increasing legume proportion increased propionate concentration (P = 0.03). Regardless of TF-legume ratio, feeding TF+CM re sulted in the greatest propionate concentration, whereas TF+AF and TF+BT maintained a similar concentration of propionate. The TF+AF combination resulted in a greater acetate-to-propionate ratio than TF+BT or TF+CM (P = 0.03). Decrease in ammonia-N concentration (P

Comparative activities of selected fluoroquinolones against dynamic populations of Actinobacillus pleuropneumoniae in an in vitro model of time–kill continuous culture experiment

The aim of the current study was to demonstrate and compare the impact of different pharmacokinetics of marbofloxacin, enrofloxacin and difloxacin on their antimicrobial effects, their killing and re-growth kinetics, and the population dynamics of Actinobacillus pleuropneumoniae clinical isolates in an in vitro dynamic model. Selected clinical isolates of A. pleuropneumoniae and three fluoroquinolones at a range of simulated AUC24/MIC ratios of multiple doses were investigated. At the same simulated AUC24/MIC ratios of the three fluoroquinolones, the killing re-growth profile and IE values (intensity of the antimicrobial effect) revealed strain- and fluoroquinolone-specific effects. For example, a 31% lower IE of difloxacin was observed in AppK5 (biofilm-former) than in AppK2 (biofilm-non-former) at the same AUC24/MIC ratio of 120h. In addition, losses in A. pleuropneumoniae susceptibility of both strains by the three fluoroquinolones were observed. AUC24/MPC ratios of 20.89 and 39.81 for marbofloxacin, 17.32 and 19.49 for enrofloxacin and 31.62 and 60.25 for difloxacin were estimated to be protective against the selection of AppK2 and AppK5 strain mutants, respectively. Integration of these in vitro data with published pharmacokinetics revealed the inadequacy of the conventional clinical doses of the three drugs to attain the above protective values for minimum biofilm eradication concentration (MBEC) and concentration to prevent growth of 90% of the mutant subpopulation (MPC90). In conclusion, the results suggest optimising doses could suffice for resistant mutants control, while for biofilm-forming strains combination with biofilm-disrupting agents to reduce the MBEC to achieve AUC/MBEC ratios within the possible dosing regimens is desired.

Changes in bacterial diversity in response to dissolved organic matter supply in a continuous culture experiment

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 69:157-168 (2013) - DOI: https://doi.org/10.3354/ame01632 Changes in bacterial diversity in response to dissolved organic matter supply in a continuous culture experiment Marine Landa1,2, Matthew T. Cottrell3, David L. Kirchman3, Stéphane Blain1,2, Ingrid Obernosterer1,2,* 1UPMC Univ Paris 06, UMR 7621 and 2CNRS, UMR 7621, LOMIC, Observatoire Océanologique, Banyuls-sur-Mer 66650, France 3School of Marine Science and Policy, University of Delaware, Lewes, Delaware 19958, USA *‑Corresponding author. Email: ingrid.obernosterer@obs-banyuls.fr ABSTRACT: Heterotrophic bacterial metabolism is tightly linked to the availability of dissolved organic matter (DOM), but the relationship between substrate supply and bacterial diversity remains unresolved. The objective of the present study was to determine how an increase in the supply of bioavailable DOM affects bacterial metabolism and diversity. We addressed this issue by growing a natural bacterial community over 5 generations in continuous cultures either on seawater (dissolved organic carbon concentration equal to 74 µM) or on seawater amended with 13 µM of diatom-derived DOM. Bacteria markedly responded to the additional DOM supply, as revealed by 1.6-fold higher bacterial abundances and 10-fold higher cell-specific α- and β-glucosidase activities in the +DOM treatment than in the control throughout the experimental period. Analysis of diversity as determined by tag pyrosequencing of 16S rRNA genes after 3 and 5 bacterial generations showed that the bacterial communities in the +DOM treatment were more similar to each other than to communities in the control. Diversity indices such as richness and phylogenetic distance were higher in the DOM-amended cultures than in the control. Gammaproteobacteria dominated the communities selected in our cultures, representing between 87 and 99% of the sequences. The addition of DOM led to a higher relative abundance of several bacterial groups, in particular Alphaproteobacteria, Bacteroidetes, and Verrucomicrobia, which accounted for up to 14% of the sequences, compared to the control in which they represented <5% of the sequences. Our results demonstrate that in this experimental context, an increased supply of bioavailable DOM sustained a higher bacterial diversity. KEY WORDS: Bacterial diversity · Dissolved organic matter · Continuous cultures · 454 pyrosequencing Full text in pdf format Supplementary material PreviousNextCite this article as: Landa M, Cottrell MT, Kirchman DL, Blain S, Obernosterer I (2013) Changes in bacterial diversity in response to dissolved organic matter supply in a continuous culture experiment. Aquat Microb Ecol 69:157-168. https://doi.org/10.3354/ame01632 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 69, No. 2. Online publication date: May 28, 2013 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2013 Inter-Research.

Rumen degradable protein supply affects microbial efficiency in continuous culture and growth in steers

We hypothesized that microbial efficiency and output from fermentation in the rumen would be optimized when peptide supply was balanced with peptide requirement of ruminal microflora. This study was conducted to measure response of varying rumen degradable peptide (RDPep) supply on ruminal fermentation characteristics and steer growth. A continuous culture experiment was conducted with diets formulated to achieve a predicted RDPep balance (RDPep supplied above RDPep required) of -0.30 to 1.45% CP with rumen degradable N (RDN) balance (RDN supplied above RDN required) above dietary ammonia-N requirement of microbes. Two additional treatments had RDPep balances of -0.30 and 0.78% CP with insufficient ammonia-N supply to meet microbial requirements. Single-flow fermenters (N = 24; n = 6) were inoculated with rumen fluid and maintained anaerobically at 39°C with a 0.06 h(-1) dilution rate. Inadequate RDN decreased OM digestion and microbial N flow, and increased rumen undegradable N (P < 0.01). Microbial efficiency decreased in RDN-deficient diets and was greatest when RDPep balance did not excessively exceed microbial requirement of RDPep predicted (P < 0.01). A growth study was conducted with 49 yearling, crossbred, Angus steers (initial BW 370 ± 34 kg). Animals were assigned to 1 of 4 treatment groups by BW and further divided into 3 pens with 4 steers per pen to achieve similar initial pen weights. Treatments consisted of 4 isonitrogenous diets balanced for RDN but varying in predicted RDPep balance (0.55%, -0.02%, -0.25%, and -0.65% CP). Animals were maintained on treatment for 70 d with individual BW taken on d 0, 1, 21, 42, 70, and 71. Final BW decreased linearly with decreasing RDPep (P = 0.05). Average daily gain and G:F displayed a quadratic effect with greater ADG and G:F at greater and lesser RDPep levels (P = 0.02). We concluded that balancing RDPep supply to predicted requirement improved fermentation efficiency and microbial output, which in turn improved animal performance.

Effect of dietary crude glycerol level on ruminal fermentation in continuous culture and growth performance of beef calves

Continuous culture and in vivo experiments were conducted to measure changes in ruminal fermentation and animal performance when crude glycerol was added to diets. For the continuous culture experiment (n = 6), diets consisted of 4 levels of crude glycerol (0, 5, 10, and 20%) that replaced corn grain. Dry matter and OM digestibility decreased linearly (P < 0.05) when crude glycerol increased in the diet, and no effect (P = 0.20 and 0.65, respectively) was observed for CP and NDF digestibility. Total VFA concentration and ammonia did not change (P > 0.05) due to crude glycerol level. Microbial efficiency increased quadratically (P = 0.012) as crude glycerol increased, whereas microbial N flow did not differ (P = 0.36) among treatments. As crude glycerol increased in the diet, crude glycerol digestibility decreased (P < 0.05). Seventy-two crossbred steer calves (250 ± 2.0 kg) were assigned to 4 treatments: 0, 5, 10, and 20% crude glycerol that replaced corn grain. Animals were fed for a total of 150 d. No differences (P = 0.08) between treatments were measured for DMI. Average daily gain and GF responded quadratically (P < 0.05), with 10% crude glycerol resulting in the greatest values. In the second in vivo experiment, 100 crossbred steer calves (300 ± 2.0 kg) were assigned to 5 treatments: 0, 5, 10, 12.5, or 15% crude glycerol replaced corn grain. Calves were fed for a total of 135 d. No significant differences (P > 0.05) were measured in growth performance. For Exp. 3, one hundred heifer calves (270 ± 2.0 kg) were assigned to 4 treatments: 0, 5, 10, or 20% crude glycerol that replaced hay. No differences (P > 0.05) were measured in animal performance. We concluded that crude glycerol addition to a diet did not negatively affect ruminal fermentation, and addition of up to 20% in concentrate and hay-based diets should not affect performance or carcass characteristics.

Interactive effects of iron, irradiance and CO 2 on Ross Sea phytoplankton

We conducted a factorial shipboard continuous culture experiment to examine the interactive effects of altered iron, irradiance and CO 2 on the summer phytoplankton community of the Ross Sea, Antarctica. After 18 days of continuous incubation, iron enrichment increased phytoplankton biomass, nutrient drawdown, diatom and Phaeocystis abundance, and some photosynthetic parameters. High irradiance significantly increased the number of Phaeocystis antarctica colonies, as well as P. antarctica abundance relative to diatoms. Iron and light had significant interactive effects on diatom and P. antarctica pigment concentrations, P. antarctica colony abundance, and Si:N, Si:C, and N:P ratios. The major influence of high CO 2 was on diatom community structure, by favoring the large centric diatom Chaetoceros lineola over the small pennate species Cylindrotheca closterium. The ratio of centric to pennate diatoms was significantly responsive to changes in all three variables individually, and to all of their possible two- and three-way combinations. These results suggest that shifts in light, iron, and CO 2 and their mutual interactions all play a role in controlling present day Ross Sea plankton community structure, and need to be considered when predicting the possible future responses of biology and biogeochemistry in this region.

Effects of increased pCO2 and temperature on the North Atlantic spring bloom. I. The phytoplankton community and biogeochemical response

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 388:13-25 (2009) - DOI: https://doi.org/10.3354/meps08133 Effects of increased pCO2 and temperature on the North Atlantic spring bloom. I. The phytoplankton community and biogeochemical response Yuanyuan Feng1,8, Clinton E. Hare1, Karine Leblanc1,9,10, Julie M. Rose1,11, Yaohong Zhang1, Giacomo R. DiTullio2, Peter A. Lee2, Steven W. Wilhelm3, Janet M. Rowe3,12, Jun Sun4, Nina Nemcek5, Celine Gueguen5,13, Uta Passow6, Ina Benner6, Christopher Brown7, David A. Hutchins1,8,* 1College of Marine and Earth Studies, University of Delaware, 700 Pilottown Road, Lewes, Delaware 19958, USA 2Hollings Marine Laboratory, College of Charleston, 331 Fort Johnson Road, Charleston, South Carolina 29412, USA 3Department of Microbiology, University of Tennessee, 1414 West Cumberland Ave, Knoxville, Tennessee 37996, USA 4Key Laboratory of Marine Ecology and Environmental Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, PR China 5Department of Earth and Ocean Science, The University of British Columbia, 6339 Stores Road, Vancouver, British Columbia V6T 1Z4, Canada 6Alfred Wegener Institute for Polar and Marine Research, Am Handelshafen 12, Bremerhaven 27568, Germany 7Earth System Science Interdisciplinary Center, University of Maryland Research Park (M-Square), 5825 University Research Ct, Ste 4001, College Park, Maryland 20740, USA 8Present address: Department of Biological Sciences, University of Southern California, 3616 Trousdale Parkway, Los Angeles, California 90089, USA 9Present address: Aix-Marseille Université, CNRS, LOB-UMR 6535, Laboratoire d’Océanographie et de Biogéochimie, OSU/Centre d’Océanologie de Marseille, 163 Avenue de Luminy, 13288 Marseille Cedex 09, France 10Present address: CNRS (CNRS/INSU), UMR 6535, Campus de Luminy Case 901, 163 Avenue de Luminy, 13288 Marseille Cedex 09, France 11Present address: Biology Department, MS #32, Woods Hole Oceanographic Institution, 266 Woods Hole Road, Woods Hole, Massachusetts 02543, USA 12Present address: Department of Biological Sciences, The University of Nebraska, 204 Morrison Center Lincoln, Nebraska 68583-0900, USA 13Present address: Department of Chemistry, Trent University, 1600 West Bank Drive, Peterborough, Ontario K9J 7B8, Canada *Corresponding author. Email: dahutch@usc.edu ABSTRACT: The North Atlantic spring bloom is one of the largest annual biological events in the ocean, and is characterized by dominance transitions from siliceous (diatoms) to calcareous (coccolithophores) algal groups. To study the effects of future global change on these phytoplankton and the biogeochemical cycles they mediate, a shipboard continuous culture experiment (Ecostat) was conducted in June 2005 during this transition period. Four treatments were examined: (1) 12°C and 390 ppm CO2 (ambient control), (2) 12°C and 690 ppm CO2 (high pCO2), (3) 16°C and 390 ppm CO2 (high temperature), and (4) 16°C and 690 ppm CO2 (‘greenhouse’). Nutrient availability in all treatments was designed to reproduce the low silicate conditions typical of this late stage of the bloom. Both elevated pCO2 and temperature resulted in changes in phytoplankton community structure. Increased temperature promoted whole community photosynthesis and particulate organic carbon (POC) production rates per unit chlorophyll a. Despite much higher coccolithophore abundance in the greenhouse treatment, particulate inorganic carbon production (calcification) was significantly decreased by the combination of increased pCO2 and temperature. Our experiments suggest that future trends during the bloom could include greatly reduced export of calcium carbonate relative to POC, thus providing a potential negative feedback to atmospheric CO2 concentration. Other trends with potential climate feedback effects include decreased community biogenic silica to POC ratios at higher temperature. These shipboard experiments suggest the need to examine whether future pCO2 and temperature increases on longer decadal timescales will similarly alter the biological and biogeochemical dynamics of the North Atlantic spring bloom. KEY WORDS: Ocean acidification · Global change · Carbon dioxide · Temperature · Coccolithophores · Diatoms · Calcification · North Atlantic bloom Full text in pdf format PreviousNextCite this article as: Feng Y, Hare CE, Leblanc K, Rose JM and others (2009) Effects of increased pCO2 and temperature on the North Atlantic spring bloom. I. The phytoplankton community and biogeochemical response. Mar Ecol Prog Ser 388:13-25. https://doi.org/10.3354/meps08133 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 388. Online publication date: August 19, 2009 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2009 Inter-Research.

Biodiversity and dye decolourization ability of an acclimatized textile sludge

In the present study, sludge sample from biological treatment plant of a textile industry was acclimatized for decolourization of azo dye Direct Black 38 (DB38). A continuous culture experiment showed that the acclimatized sludge could decolourize 76% of 100 mg/l DB38. Bacterial community in the sludge was analyzed using culture-independent molecular approach to get the complete picture of its diversity. RFLP analysis of its 16S rRNA gene library divided the clones into 14 distinct groups. Phylogenetic analysis of these groups showed that they belonged to five different bacterial lineages: β - and γ -Proteobacteria (3 and 4 respectively), Bacteroidetes (2), Firmicutes (4) and Actinobacteria (1). The largest number of clones was found to cluster in the γ -Proteobacteria (54%), followed by Firmicutes (19%), β -Proteobacteria (14%), Bacteroidetes (10%) and Actinobacteria (3%).

Interactions of algal ligands, metal complexation and availability, and cell responses of the diatom Ditylum brightwellii with a gradual increase in copper

A continuous culture experiment was conducted to study interactions between copper-binding ligands released by light-limited Ditylum brightwellii, and toxic effects of Cu on this diatom. Over 6 months, the Cu concentration in the medium has been increased in seven steps (3–173 nM). At each Cu addition, Cu speciation, characteristics of Cu sorption to cellular binding sites, and cell characteristics were determined. Physiological effects of Cu were studied, using indicators for metal detoxification (thiols) and lipid peroxidation (malondialdehyde). Minor amounts of Cu (<1.4%) were chelated by a minimum amount of EDTA (57 nM), required to maintain a stable long-term continuous culture. The responses of D. brightwellii to Cu were monitored. (1) From 3 to 47 nM added Cu, decreasing pools of glutathione, increasing malondialdehyde contents, an increased release of lipophilic ligands, and cell lysis indicated the enhancement of lipid peroxidation. (2) From 47 to 94 nM Cu, a 16-fold increase in high-affinity (strong) hydrophilic ligands was measured (conditional stability constants K′≈10 12) that complexed most Cu (maximum 97%); sexual reproduction was stimulated and cell volumes increased. (3) From 126 nM Cu, glutathione pools increased again, whereas cell division rates decreased slightly. (4) At 142 nM Cu, the number of lysed cells reached a maximum, as did the production of lipophilic compounds that complexed ∼2% Cu. As the binding sites of the strong ligands became Cu-saturated above 142 nM Cu, larger amounts of Cu were bound to low-affinity (weak) dissolved ligands (3–30%) and cellular binding sites (0.2–2.5%). Probably due to saturation of organic complexes at 142 nM Cu, the MINEQL-calculated Cu 2+ concentrations increased markedly; pCu values decreased from >11 to ∼10; division rates were further inhibited; γ-glutamylcysteine (phytochelatin precursor) was produced. (5) At 157 nM Cu, phytochelatin synthesis started, and Cu-sorption capacities (cell walls and internal binding sites) increased. (6) At 173 nM Cu, the phytochelatin pool sizes and the number of cellular Cu-binding sites increased further. These results suggest that ligands released by a dense bloom of D. brightwellii, either by active excretion or lysis, would have lower affinities for Cu ( K′≈10 9–10 12) and moderate the availability of Cu less effectively than ligands in natural environments (10 13–10 14). In this diatom, the concurring release of ligands, enhanced malondialdehyde production, increasing numbers of presexual cells and cell enlargement may serve as early-warning signals for Cu toxicity, rather than metal-specific phytochelatins that appeared at a stage when cell division was already clearly inhibited.

Effects of periodic, low UVA radiation on cell characteristics and oxidative stress in the marine planktonic diatom Ditylum brightwellii

A continuous culture experiment was conducted with the marine diatom Ditylum brighiwellii to assess the photo-oxidative effects of transition from dim light to high Light at three successive levels. The focus was on temporary stress by ambient WA at the water surface, via a simulation of the periodic ascent of diatoms caused by vertical mixing. Within a 14 h dim- light period, cells were exposed daily to 4 h of: (A) moderate PAR (100 mu mol photon m(-2) s(-1)), (B) high PAR (400 mu mol photon m(-2) s(-1)) and (C) high PAR (400-700 nm) plus 67 pmol photon m(-2) s(-1) (2.2 W m(-2)) UVA. Cell shape, life cycle and sinking rates were not affected by high PAR per se (mode B). The daily 4 h high-irradiance mode C, with additional UVA (WA:PAR = 0.016), caused an increase in cell diameter, a decrease in length axes, and a production of vegetative resting stages. Superoxide dismutase (SOD) activities, oxidized glutathione (GSSG) pools and malondialdehyde (MDA) contents increased as well, which indicates that this biologically effective WA dose (0.78 W m(-2)) promoted active oxygen production, oxidative stress and lipid peroxidation. Sinking rates in WA-exposed D. brightwellii were higher than in those grown without UVA. A close correlation between sinking rates and MDA contents suggests that UVA-induced lipid peroxidation has altered the membrane functions which regulate buoyancy in D. brightwellii. Although UVA-induced formation of resting cells will contribute to accelerated sinking, it is not evident that this can be considered as a stress avoidance mechanism under adverse light (WA) conditions. D. brightwellii proved to be sensitive to transitions from shade to natural surface irradiance conditions. In coastal waters, such large diatom species may experience oxidative stress after a few hours' residence near the water surface at ambient UVA levels. [KEYWORDS: cell size; Ditylum brightwellii; glutathione redox;lipid peroxidation; oxidative stress; planktonic diatom; resting stage; sinking rate; superoxide; ultraviolet-A Ultraviolet-b radiation; lipid-peroxidation; chlorella-vulgaris; photosynthetic pigment; superoxide-dismutase; thylakoid membranes; antioxidant systems; maize seedlings; sinking rates; phytoplankton]

Effects of periodic, low UVA radiation on cell characteristics and oxidative stress in the marine planktonic diatom Ditylum brightwellii

A continuous culture experiment was conducted with the marine diatom Ditylum brightwellii to assess the photo-oxidative effects of transition from dim light to high light at three successive levels. The focus was on temporary stress by ambient UVA at the water surface, via a simulation of the periodic ascent of diatoms caused by vertical mixing. Within a 14 h dim-light period, cells were exposed daily to 4 h of : (A) moderate PAR (100 μmol photon m−2 s−1), (B) high PAR (400 μmol photon m−2 s−1) and (C) high PAR (400–700 nm) plus 6·7 μmol photon m−2 s−1 (2·2 W m−2) UVA. Cell shape, life cycle and sinking rates were not affected by high PAR per se (mode B). The daily 4 h high-irradiance mode C, with additional UVA (UVA:PAR = 0·016), caused an increase in cell diameter, a decrease in length axes, and a production of vegetative resting stages. Superoxide dismutase (SOD) activities, oxidized glutathione (GSSG) pools and malondialdehyde (MDA) contents increased as well, which indicates that this biologically effective UVA dose (0·78 W m−2) promoted active oxygen production, oxidative stress and lipid peroxidation. Sinking rates in UVA-exposed D. brightwellii were higher than in those grown without UVA. A close correlation between sinking rates and MDA contents suggests that UVA-induced lipid peroxidation has altered the membrane functions which regulate buoyancy in D. brightwellii. Although UVA-induced formation of resting cells will contribute to accelerated sinking, it is not evident that this can be considered as a stress avoidance mechanism under adverse light (UVA) conditions. D. brightwellii proved to be sensitive to transitions from shade to natural surface irradiance conditions. In coastal waters, such large diatom species may experience oxidative stress after a few hours' residence near the water surface at ambient UVA levels.

Ruminal peptide concentration required to optimize microbial growth and efficiency.

Experimentation, with a single-phase continuous culture system operated at fractional dilution rates of 0.03 and 0.09 per hour and four cannulated crossbred steers (260 +/- 20 kg) used in a 4 x 4 Latin square design, was conducted to determine the level of ruminally degradable protein (RDP) that maximized microbial efficiency (MOEFF), microbial nitrogen flow, and nutrient digestibility in the rumen. Treatments consisted of increasing diet RDP levels (from 3.5 to 17.5% RDP on a DM basis). The basal diet was corn with casein or soybean meal used as the source of RDP in the continuous culture experiment and animal study, respectively. Dietary nonstructural carbohydrate (NSC) ranged from 52 to 75% on a dry matter basis. Urea was added to ensure an adequate ammonia nitrogen source for microbial growth. The RDP levels did not affect true digestibility of dry matter and organic matter, ruminal pH, particulate passage rate, or liquid passage rate in the animal study. As RDP increased, bacterial nitrogen production increased linearly (P < 0.05) only for the 0.09/h dilution rate. Microbial efficiency was not influenced by RDP level. Ruminal peptide and NH3 N concentration increased linearly (P < 0.05) as RDP increased. Based on this experimentation, 1.8 mM peptide maximized MOEFF when ammonia nitrogen was not limiting (> 2 mg/dL). We concluded from this research that the RDP requirement of NSC-fermenting bacteria was lower than that currently fed in many diets.

Use of chemostat for enhanced production of β-glucosidase by newly isolated anaerobic cellulolyticClostridium strain RT9

A new anaerobic, mesophilic, spore-forming, cellulolyticClostridium strain, RT9, was isolated from bovine rumen fluid. This strain has the ability to hydrolyze cellulose (Sigma cell-100), carboxymethyl-cellulose (CMC), and cellobiose into ethanol, acetate, butyrate, lactate, acetone, H2, and Co2 as the major fermentation products. The bacterium also exhibited endoglucanase, exoglucanase, and β-glucosidase activities in batch culture during growth on chemically defined medium with cellulose as the sole carbon source. Chemostat experiments were conducted under pH, dilution rate, and carbohydrate limitations to study the effect of various parameters on enzymes and product formation. In chemostat with 0.5% cellobiose as the limiting nutrient, maximum β-glucosidase activity and ethanol production occurred at pH values of 5.5 and 6.5, respectively. At low pH, acid accumulation was higher. β-glucosidase production increased up to 17-fold at a low dilution rate, and ethanol raised twofold. Relative activity of β-glucosidase on a percent basis was maximum at pH 6.5 and 45 °C. In a carbon-limited, continuous-culture experiment, when CMC was used as an inducer in cellobiose-growing culture, β-glucosidase activity was higher than the cellobiose-limited chemostat. On the other hand, enzyme declined twofold, and ethanol production increased in glucose limitation.

Recombinant protein synthesis and plasmid instability in continuous cultures of Escherichia coli JM103 harboring a high copy number plasmid

Escherichia coli JM103[pUC8] was employed as a model to investigate the behavior of a recombinant microbial system harboring a plasmid at high copy numbers. Experiments with batch and continuous cultures of recombinant and plasmid-free cells were conducted in a well-controlled bio-reactor. In batch experiments, plasmid copy number varied typically from an average of 500 during the exponential growth phase to as high as 1250 during the stationary phase. While the segregational plasmid instability was negligible in batch experiments, severe segregational instability occurred in continuous experiments conducted over a range of dilution rates, resulting in complete loss of plasmid-bearing cells from the continuous cultures within few residence times after transition to continuous operation. The profound differences in the specific growth rates and mass yields of the plasmid-free and plasmid-bearing cells resulting from the extra metabolic burden on the plasmid-bearing cells mainly due to excessive plasmid DNA content was the major cause for the plasmid instability. Plasmid multirnerization was detected in batch and continuous cultures and was found to have significant influence on the effective copy number and was partially responsible for the severe segregational instability in continuous cultures. A quasi-steady state representative of plasmid-bearing cells was established in the initial portion of each continuous culture experiment. Due to the profound growth rate differential between the two types of cells, transients of considerable duration were observed in each continuous culture experiment (initiated with a pure culture of plasmid bearing cells) following the slow accumulation of plasmid-free cells near the end of the quasi-steady state. Significant variations in various culture parameters (including a rapid decline in the plasmid-bearing fraction of the total cell population) occurred during this period, leading ultimately to a steady state for a culture dominated entirely by plasmid-free cells. In continuous cultures, plasmid copy number during the quasi-steady states increased with decreasing dilution rate from 50 (at 0.409 h(-1)) to 941 (at 0.233 h(-1)). Production of the plasmid-encoded protein (beta-lactamase) in these experiments was maximized at an intermediate dilution rate, corresponding to an optimum copy number of about 450. A similar optimum copy number was observed in batch cultures. Significant excretion of beta-lactamase was observed at both low and high dilution rates.