Copper enhances the activity and salt resistance of mixed methane-oxidizing communities

Effluents of anaerobic digesters are an underestimated source of greenhouse gases, as they are often saturated with methane. A post-treatment with methane-oxidizing bacterial consortia could mitigate diffuse emissions at such sites. Semi-continuously fed stirred reactors were used as model systems to characterize the influence of the key parameters on the activity of these mixed methanotrophic communities. The addition of 140 mg L⁻¹ NH₄⁺–N had no significant influence on the activity nor did a temperature increase from 28°C to 35°C. On the other hand, addition of 0.64 mg L⁻¹ of copper(II) increased the methane removal rate by a factor of 1.5 to 1.7 since the activity of particulate methane monooxygenase was enhanced. The influence of different concentrations of NaCl was also tested, as effluents of anaerobic digesters often contain salt levels up to 10 g NaCl L⁻¹. At a concentration of 11 g NaCl L⁻¹, almost no methane-oxidizing activity was observed in the reactors without copper addition. Yet, reactors with copper addition exhibited a sustained activity in the presence of NaCl. A colorimetric test based on naphthalene oxidation showed that soluble methane monooxygenase was inhibited by copper, suggesting that the particulate methane monooxygenase was the active enzyme and thus more salt resistant. The results obtained demonstrate that the treatment of methane-saturated effluents, even those with increased ammonium (up to 140 mg L⁻¹ NH₄⁺–N) and salt levels, can be mitigated by implementation of methane-oxidizing microbial consortia.     

Productivity correlated to photobiochemical performance of <Emphasis Type="Italic">Chlorella</Emphasis> mass cultures grown outdoors in thin-layer cascades

This work aims to: (1) correlate photochemical activity and productivity, (2) characterize the flow pattern of culture layers and (3) determine a range of biomass densities for high productivity of the freshwater microalga Chlorella spp., grown outdoors in thin-layer cascade units. Biomass density, irradiance inside culture, pigment content and productivity were measured in the microalgae cultures. Chlorophyll-fluorescence quenching was monitored in situ (using saturation-pulse method) to estimate photochemical activities. Photobiochemical activities and growth parameters were studied in cultures of biomass density between 1 and 47 g L⁻¹. Fluorescence measurements showed that diluted cultures (1–2 g DW L⁻¹) experienced significant photostress due to inhibition of electron transport in the PSII complex. The highest photochemical activities were achieved in cultures of 6.5–12.5 g DW L⁻¹, which gave a maximum daylight productivity of up to 55 g dry biomass m⁻² day⁻¹. A midday depression of maximum PSII photochemical yield (F _v/F _m) of 20–30% compared with morning values in these cultures proved to be compatible with well-performing cultures. Lower or higher depression of F _v/F _m indicated low-light acclimated or photoinhibited cultures, respectively. A hydrodynamic model of the culture demonstrated highly turbulent flow allowing rapid light/dark cycles (with frequency of 0.5 s⁻¹) which possibly match the turnover of the photosynthetic apparatus. These results are important from a biotechnological point of view for optimisation of growth of outdoor microalgae mass cultures under various climatic conditions.     

Transformation of indole and quinoline by Desulfobacterium indolicum (DSM 3383)

Degradation of indole and quinoline by Desulfobacterium␣indolicum was studied in batch cultures. The first step in the degradation pathway of indole and quinoline was a hydroxylation at the 2 position to oxindole and 2-hydroxyquinoline respectively. These hydroxylation reactions followed saturation kinetics. The kinetic parameters for indole were an apparent maximum specific transformation rate (V _Amax) of 263 μmol mg total protein⁻¹ day⁻¹ and an apparent half-saturation constant (K _Am) of 139 μM. The V _Amax for quinoline was 170 μmol mg total protein⁻¹ day⁻¹ and K _Am was 92 μM. Oxindole inhibited indole hydroxylation whereas 2-hydroxyquinoline stimulated quinoline hydroxylation. An adaptation period of approximately 20 days was required before transformation of 2-hydroxyquinoline in cultures previously grown on quinoline. Indole and quinoline were hydroxylated with a lag phase shorter than 4 h in a culture adapted to ethanol. Chloramphenicol inhibited the hydroxylation of indole and quinoline in ethanol-adapted cells, indicating an inducible enzyme system. Chloramphenicol had no effect on the hydroxylation of indole in quinoline-adapted cells or on the hydroxylation of quinoline in indole-adapted cells. This indicated that it was the same inducible enzyme system that hydroxylated indole and quinoline. Received: 16 July 1996 / Received revision: 23 September 1996 / Accepted: 29 September 1996     

In vitro and in vivo evaluation of the prebiotic activity of water-soluble blueberry extracts

The prebiotic effects of water extracts of two blueberry (BBE) cultivars (‘Centurion’ and ‘Maru’) were studied using pure and mixed cultures of human faecal bacteria. The results demonstrated for the first time that addition of BBE from both cultivars to broth media containing pure cultures of Lactobacillus rhamnosus and Bifidobacterium breve resulted in a significant increase (P < 0.05–0.0001) in the population size of these strains. Batch fermentation system was used to monitor the effect of BBE addition on the mixed faecal bacterial populations (obtained from healthy human donors). Addition of BBE from both cultivars to batch cultures inoculated with mixed human faecal cultures resulted in a significant increase in the number of lactobacilli (P < 0.01–0.0001) and bifidobacteria (P < 0.05–0.0001). Furthermore, a significant influence on the population size of lactobacilli and bifidobacteria was observed after administration of extracts from both cultivars to rats daily for 6 days in comparison with the control group. In rats gavaged orally with 4 ml kg⁻¹ day⁻¹ of BBE for 6 days, the population size of lactobacilli (P < 0.05) and bifidobacteria (P < 0.05–0.01) was increased significantly. We hypothesize that BBE could modify the bacterial profile by increasing the numbers of beneficial bacteria and thereby improving gut health.     

Nitrogen retention in the hyporheic zone of a glacial river in interior Alaska

We examined the hydrologic controls on nitrogen biogeochemistry in the hyporheic zone of the Tanana River, a glacially-fed river, in interior Alaska. We measured hyporheic solute concentrations, gas partial pressures, water table height, and flow rates along subsurface flowpaths on two islands for three summers. Denitrification was quantified using an in situ ¹⁵NO₃⁻ push–pull technique. Hyporheic water level responded rapidly to change in river stage, with the sites flooding periodically in mid−July to early−August. Nitrate concentration was nearly 3-fold greater in river (ca. 100 μg NO₃⁻–N l⁻¹) than hyporheic water (ca. 38 μg NO₃⁻–N l⁻¹), but approximately 60–80% of river nitrate was removed during the first 50 m of hyporheic flowpath. Denitrification during high river stage ranged from 1.9 to 29.4 mg N kg sediment⁻¹ day⁻¹. Hotspots of methane partial pressure, averaging 50,000 ppmv, occurred in densely vegetated sites in conjunction with mean oxygen concentration below 0.5 mgO₂ l⁻¹. Hyporheic flow was an important mechanism of nitrogen supply to microbes and plant roots, transporting on average 0.41 gNO₃⁻–N m⁻² day⁻¹, 0.22 g NH₄⁺–N m⁻² day⁻¹, and 3.6 g DON m⁻² day⁻¹ through surface sediment (top 2 m). Our results suggest that denitrification can be a major sink for river nitrate in boreal forest floodplain soils, particularly at the river-sediment interface. The stability of the river hydrograph and the resulting duration of soil saturation are key factors regulating the redox environment and anaerobic metabolism in the hyporheic zone.     

Studies on non-symbiotic diazotrophic bacterial populations of coastal arable saline soils of India

The effect of fluctuations of salinity in three different seasons on diazotrophic populations and N₂ fixation in six mono cropped rice field soils of the coastal region of the Gangetic delta of West Bengal, India, was studied. The average pH, ECe, organic carbon and total nitrogen of the soils ranged from 4.99–7.08, 2.02–19.58 dSm⁻¹, 4.68–12.03 g kg⁻¹ and 0.44–1.70 g kg ⁻¹, respectively. The average log colony forming units of the bacterial populations and N₂-fixation in the soils varied from 4.61 to 5.86 and 2.74 to 4.52 mg N₂ fixed 50 ml ⁻¹ culture media respectively, with the lowest value recorded in summer. Recovery of microorganisms and N₂- fixation gradually decreased with extraneous addition of NaCl in the culture media. All the eight isolates were Gram positive, spore and capsule formers. They could utilize glucose, sucrose, mannitol, starch, citrate and nitrate, and were catalase and gelatinase positive, but indole, methyl red and Vogues Proskauer reaction negative. The organisms produced alkaline reaction on TSI agar slant. The acetylene reduction assay of the isolates at 0 and 1% NaCl in the culture media were 4.51–164.52 and 1.72–100.6 nmole C₂H₄ ml⁻¹ culture media in 72 h, respectively. The isolates could fix 2.42–4.45 and 2.04–4.08 mg N₂ fixed 50 ml⁻¹ culture media at 0 and 1% NaCl in the culture media respectively. 16S rDNA sequences of the isolates were similar to the species: Bacillus sp. isolate 28A, Bacillus sp. MOLA 87, Bacillus sp. By113 (B)Ydz-dh, Bacillus sp. PN13, Bacillus licheniformis strain RH101, Bacterium Antarctica 14, Bacillus sp. PN13 and Bacillus megaterium.     

Biofiltering efficiency, uptake and assimilation rates of Ulva clathrata (Roth) J. Agardh (Clorophyceae) cultivated in shrimp aquaculture waste water

The growth, biofiltering efficiency and uptake rates of Ulva clathrata were studied in a series of outdoor tanks, receiving waste water directly from a shrimp (Litopenaeus vannamei) aquaculture pond, under constant aeration and two different water regimes: (1) continuous flow, with 1 volume exchange a day (VE day^-1) and (2) static regime, with 1 VE after 4 days. Water temperature, salinity, pH, dissolved inorganic nitrogen (DIN), phosphate (PO₄), chlorophyll-a (chl-a), total suspended solids (TSS), macroalgal biomass (fresh weight) and tissue nutrient assimilation were monitored over 12 days. Ulva clathrata was highly efficient in removing the main inorganic nutrients from effluent water, stripping 70–82% of the total ammonium nitrogen (TAN) and 50% PO₄ within 15 h. Reductions in control tanks were much lower (Tukey HSD, P < 0.05). After 3 days, the mean uptake rates by the seaweed biomass under continuous flow were 3.09 mg DIN g DW day⁻¹ (383 mg DIN m⁻² day⁻¹) and 0.13 mg PO₄ g DW day⁻¹ (99 mg PO₄ m⁻² day⁻¹), being significantly higher than in the static regime (Tukey HSD, P < 0.05). The chl-a decreased in seaweed tanks, suggesting that U. clathrata inhibited phytoplankton growth. Correlations between the cumulative values of DIN removed from the water and total nitrogen assimilated into the seaweed biomass (r = 0.7 and 0.8, P < 0.05), suggest that nutrient removal by U. clathrata dominated over other processes such as phytoplankton and bacterial assimilation, ammonia volatilization and nutrient precipitation.     

Production of Dissolved Organic Carbon in Canadian Forest Soils

To identify the controls on dissolved organic carbon (DOC) production, we incubated soils from 18 sites, a mixture of 52 forest floor and peats and 41 upper mineral soil samples, at three temperatures (3, 10, and 22°C) for over a year and measured DOC concentration in the leachate and carbon dioxide (CO₂) production from the samples. Concentrations of DOC in the leachate were in the range encountered in field soils (<2 to >50 mg l⁻¹). There was a decline in DOC production during the incubation, with initial rates averaging 0.03–0.06 mg DOC g⁻¹ soil C day⁻¹, falling to averages of 0.01 mg g⁻¹ soil C day⁻¹; the rate of decline was not strongly related to temperature. Cumulative DOC production rates over the 395 days ranged from less than 0.01 to 0.12 mg g⁻¹ soil C day⁻¹ (0.5–47.6 mg g⁻¹ soil C), with an average of 0.021 mg g⁻¹ soil C day⁻¹ (8.2 mg g⁻¹ soil C). DOC production rate was weakly related to temperature, equivalent to Q₁₀ values of 0.9 to 1.2 for mineral samples and 1.2 to 1.9 for organic samples. Rates of DOC production in the organic samples were correlated with cellulose (positively) and lignin (negatively) proportion in the organic matter, whereas in the mineral samples C and nitrogen (N) provided positive correlations. The partitioning of C released into CO₂–C and DOC showed a quotient (CO₂–C:DOC) that varied widely among the samples, from 1 to 146. The regression coefficient of CO₂–C:DOC production (log₁₀ transformed) ranged from 0.3 to 0.7, all significantly less than 1. At high rates of DOC production, a smaller proportion of CO₂ is produced. The CO₂–C:DOC quotient was dependent on incubation temperature: in the organic soil samples, the CO₂–C:DOC quotient rose from an average of 6 at 3 to 16 at 22°C and in the mineral samples the rise was from 7 to 27. The CO₂–C:DOC quotient was related to soil pH in the organic samples and C and N forms in the mineral samples.     

Utilization of <Emphasis Type="Italic">Anabaena</Emphasis> sp. in CO<Subscript>2</Subscript> removal processes

This paper focuses on modelling the growth rate and exopolysaccharides production of Anabaena sp. ATCC 33047, to be used in carbon dioxide removal and biofuels production. For this, the influence of dilution rate, irradiance and aeration rate on the biomass and exopolysaccharides productivity, as well as on the CO₂ fixation rate, have been studied. The productivity of the cultures was maximum at the highest irradiance and dilution rate assayed, resulting to 0.5 g_bio l⁻¹ day⁻¹ and 0.2 g_eps l⁻¹ day⁻¹, and the CO₂ fixation rate measured was 1.0 gCO₂ l⁻¹ day⁻¹. The results showed that although Anabaena sp. was partially photo-inhibited at irradiances higher than 1,300 μE m⁻² s⁻¹, its growth rate increases hyperbolically with the average irradiance inside the culture, and so does the specific exopolysaccharides production rate. The latter, on the other hand, decreases under high external irradiances, indicating that the exopolysaccharides metabolism hindered by photo-damage. Mathematical models that consider these phenomena have been proposed. Regarding aeration, the yield of the cultures decreased at rates over 0.5 v/v/min or when shear rates were higher than 60 s⁻¹, demonstrating the existence of thus existence of stress damage by aeration. The behaviour of the cultures has been verified outdoors in a pilot-scale airlift tubular photobioreactor. From this study it is concluded that Anabaena sp. is highly recommended to transform CO₂ into valuable products as has been proved capable of metabolizing carbon dioxide at rates of 1.2 gCO₂ l⁻¹ day⁻¹ outdoors. The adequacy of the proposed equations is demonstrated, resulting to a useful tool in the design and operation of photobioreactors using this strain.     

Oxygen consumption rate and Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase activity in early developmental stages of the sea urchin <Emphasis Type="Italic">Paracentrotus lividus</Emphasis> Lam.

Changes in oxygen consumption rate and Na⁺/K⁺-ATPase activity during early development were studied in the sea urchin Paracentrotus lividus Lam. The oxygen consumption rate increased from 0.12 μmol O₂ mg protein⁻¹ h⁻¹ in unfertilized eggs to 0.38 μmol O₂ mg protein⁻¹ h⁻¹ 25 min after fertilization. Specific activity of the Na⁺/K⁺-ATPase was significantly stimulated after fertilization, ranging up to 1.07 μmol P_i h⁻¹ mg protein⁻¹ in the late blastula stage and slightly lower values in the early and late pluteus stages.     

Influence of growth regulators and elicitors on cell growth and α-tocopherol and pigment productions in cell cultures of <Emphasis Type="Italic">Carthamus tinctorius</Emphasis> L.

The present study examined the effects of plant growth hormones, incubation period, biotic (Trametes versicolor, Mucor sp., Penicillium notatum, Rhizopus stolonifer, and Fusarium oxysporum) and abiotic (NaCl, MgSO₄, FeSO₄, ZnSO₄, and FeCl₃) elicitors on cell growth and α-tocopherol and pigment (red and yellow) productions in Carthamus tinctorius cell cultures. The cell growth and α-tocopherol and pigment contents improved significantly on Murashige and Skoog (MS) liquid medium containing 50.0 μM α-naphthalene acetic acid (NAA) and 2.5 μM 6-Benzyladenine (BA) at 28 days of incubation period. Incorporation of T. versicolor (50 mg l⁻¹) significantly enhanced the production of α-tocopherol (12.7-fold) and red pigment (4.24-fold). Similarly, supplementation of 30 mg l⁻¹ T. versicolor (7.54-fold) and 70 mg l⁻¹ Mucor sp. (7.40-fold) significantly increased the production of yellow pigment. Among abiotic elicitors, NaCl (50–70 mg l⁻¹) and MgSO₄ (10–30 mg l⁻¹) significantly improved production of α-tocopherol (1.24-fold) and red pigment (20-fold), whereas yellow pigment content increased considerably by all the abiotic elicitor treatments. Taken together, the present study reports improved productions of α-tocopherol and the pigment as a stress response of safflower cell cultures exposed to these elicitors.     

Estimation of periphytic microalgae gross primary production with DCMU-fluorescence method in Yenisei River (Siberia, Russia)

Periphyton (epilithon) gross primary production (GPP) was estimated using the DCMU-fluorescence method in the Yenisei River. In the unshaded littoral zone, chlorophyll a concentration (Chl a) and GPP value varied from 0.83 to 973.74 mg m⁻²and 2–304,425 O₂ m⁻² day⁻¹ (0.64–95 133 mg C m⁻² day⁻¹), respectively. Positive significant correlation (r = 0.8) between daily GPP and periphyton Chl a was found. Average ratio GPP:Chl a for periphyton was 36.36 mg C mg Chl a m⁻² day⁻¹. The obtained GPP values for the Yenisei River have a high significant correlation with values predicted by a conventional empirical model for stream periphyton. We concluded that the DCMU-fluorescence method can be successfully used for measuring of gross primary production of stream phytoperiphyton at least as another useful tool for such studies.     

Biomass and aboveground net primary production in a subtropical mangrove stand of Kandelia obovata (S., L.) Yong at Manko Wetland, Okinawa, Japan

Biomass and aboveground net primary production (ANPP) in a monospecific pioneer stand of a mangrove Kandelia obovata (S., L.) Yong were quantified. The estimated biomasses in leaves, branches, stems, roots, aboveground and total were 5.61 (3.68%), 28.8 (18.9%), 46.1 (30.2%), 71.8 (47.2%), 80.5 (52.8%) and 152 Mg ha⁻¹ (100%), respectively. Stem phytomass increment per tree was estimated using allometric relationships and stem analysis. Stem volume without bark of harvested trees showed a strong allometric relationship with D _0.1² H (D _0.1, diameter at a height of one-tenth of tree height H) (R ² = 0.924). Annual stem volume increment per tree showed a strong allometric relationship with D _0.1² H (R ² = 0.860). Litterfall rate ranges from 3.87 to 56.1 kg ha⁻¹ day⁻¹ for leaves and 0.177 to 46.2 kg ha⁻¹ day⁻¹ for branches. Seasonal changes of litterfall rate were observed, which showed a peak during wet season (August–September). Total annual litterfall was estimated as 10.6 Mg ha⁻¹ year⁻¹, in which 68.2% was contributed by the leaves. The ANPP in the K. obovata stand was 29.9–32.1 Mg ha⁻¹ year⁻¹, which is ca. 2.8–3.0 times of annual litterfall. The growth efficiency (aboveground biomass increment/LAI) was 5.35–5.98 Mg ha⁻¹ year⁻¹. The low leaf longevity (9.3 months) and high growth efficiency of K. obovata makes it a highly productive mangrove species.     

Simultaneous nutrients and carbon removal during pretreated swine slurry degradation in a tubular biofilm photobioreactor

The biodegradation potential of an innovative enclosed tubular biofilm photobioreactor inoculated with a Chlorella sorokiniana strain and an acclimated activated sludge consortium was evaluated under continuous illumination and increasing pretreated (centrifuged) swine slurry loading rates. This photobioreactor configuration provided simultaneous and efficient carbon, nitrogen, and phosphorous treatment in a single-stage process at sustained nitrogen and phosphorous removals efficiencies ranging from 94% to 100% and 70–90%, respectively. Maximum total organic carbon (TOC), NH₄ ⁺, and PO₄ ³⁻ removal rates of 80 ± 5 g C m_r ⁻³ day⁻¹, 89 ± 5 g N m_r ⁻³ day⁻¹, and 13 ± 3 g P m_r ⁻³ day⁻¹, respectively, were recorded at the highest swine slurry loadings (TOC of 1,247 ± 62 mg L⁻¹, N–NH₄ ⁺ of 656 ± 37 mg L⁻¹, P–PO₄ ³⁺ of 117 ± 19 mg L⁻¹, and 7 days of hydraulic retention time). The unusual substrates diffusional pathways established within the phototrophic biofilm (photosynthetic O₂ and TOC/NH₄ ⁺ diffusing from opposite sides of the biofilm) allowed both the occurrence of a simultaneous denitrification/nitrification process at the highest swine slurry loading rate and the protection of microalgae from any potential inhibitory effect mediated by the combination of high pH and high NH₃ concentrations. In addition, this biofilm-based photobioreactor supported efficient biomass retention (>92% of the biomass generated during the pretreated swine slurry biodegradation).     

Production of Spirulina sp. in sea water supplemented with anaerobic effluents in outdoor raceways under temperate climatic conditions

The use of untreated sea water supplemented with anaerobic effluents from digested pig waste and sodium bicarbonate was evaluated as a low-cost medium for semi-continuous cultivation of a mixed culture of two Spirulina strains in outdoor raceways under temperate climatic conditions (pond temperature in the range 21–26 °C and light intensity in the range 225–957␣μE m⁻² s⁻¹). The mixed culture had a predominant population (86.6 ± 3.9%) of an atypical Spirulina strain consisting of straight filaments, which appeared spontaneously after the strain with helicoidal trichomes had been subcultured. Morphological studies for the identification of the type and size of trichomes of the two strains (HF and SF) were carried out. The proportions of the two strains were observed to be stable during the monitoring period (30 days). Three different sets of semicontinuous cultures were carried out. Sets 1 and 2 were operated under regime 1 (a single addition of anaerobic effluents at time zero and no pH control) during the same season (June and July) of different years. Set 3 was operated under regime 2 (semi-continuous addition of anaerobic effluents and pH control) during the autumn. A minimum productivity of 3.6 g m⁻² day⁻¹ was obtained at one of the lowest temperatures (22.1 °C) and light intensities (245 μE m⁻² s⁻¹) and a maximum productivity of 10.9 g m⁻² day⁻¹ was observed at the highest temperature (25 °C) and highest average light intensity (618 μE m⁻² s⁻¹) registered for sets 1 and 2. The protein content in the Spirulina biomass harvested from these two sets varied from 17% to 65.6%. In set 3, a maximum productivity of 9.0 g m⁻² day⁻¹ was recorded at an average temperature of 24.4 °C and at an average light intensity of 668 μE m⁻² s⁻¹. The protein content in this set under regime 2 varied within a narrower range than in set 1 and set 2 (from 34.8% to 49.1%), apparently because of a continuous availability of ammonia nitrogen at a level of 30–50 mg l⁻¹. However, in terms of the removal of ammonia nitrogen and chemical oxygen demand, regime 1 was more efficient than regime␣2. Received: 3 September 1996 / Received revision: 19 February 1997 / Accepted: 7 March 1997     

Nitrogen and phosphorus removal through laboratory batch cultures of microalga Chlorella vulgaris and cyanobacterium Planktothrix isothrix grown as monoalgal and as co-cultures

Batch cultures of the green microalga Chlorella vulgaris and cyanobacterium Planktothrix isothrix and their corresponding co-cultures were grown in municipal wastewater in order to study their growth as well as the nitrogen (NH₄–N) and phosphorus (PO₄³⁻–P) removal. The cultures were grown under two irradiances of 20 and 60 μmol photons m⁻² s⁻¹ in shaken and unshaken conditions. The co-culture of unshaken Chlorella and Planktothrix showed the greatest growth under both irradiances. The monoalgal Planktotrix cultures showed better growth when unshaken than when shaken, whereas Chlorella cultures grew better when mixed, but only at the higher irradiance. The highest percentage of nitrogen removal (up to 80%) was attained by the unshaken co-cultures of Chlorella and Planktothrix. The amount of nitrogen recycled in the biomass reached up to 85% of that removed. Shaken monoalgal cultures of Chlorella showed phosphorus removal under both irradiances. They completely removed the initial phosphorus concentration (7.47 ± 0.17 mg L⁻¹) within 96 and 48 h under 20 and 60 μmol photons m⁻² s⁻¹, respectively.     

A process for the production of ectoine and poly(3-hydroxybutyrate) by <Emphasis Type="Italic">Halomonas boliviensis</Emphasis>

The paper reports a study involving the use of Halomonas boliviensis, a moderate halophile, for co-production of compatible solute ectoine and biopolyester poly(3-hydroxybutyrate) (PHB) in a process comprising two fed-batch cultures. Initial investigations on the growth of the organism in a medium with varying NaCl concentrations showed the highest level of intracellular accumulation of ectoine (0.74 g L⁻¹) at 10–15% (w/v) NaCl, while at 15% (w/v) NaCl, the presence of hydroxyectoine (50 mg L⁻¹) was also noted. On the other hand, the maximum cell dry weight and PHB concentration of 10 and 5.8 g L⁻¹, respectively, were obtained at 5–7.5% (w/v) NaCl. A process comprising two fed-batch cultivations was developed—the first culture aimed at obtaining high cell mass and the second for achieving high yields of ectoine and PHB. In the first fed-batch culture, H. boliviensis was grown in a medium with 4.5% (w/v) NaCl and sufficient levels of monosodium glutamate, NH₄⁺, and PO₄³⁻. In the second fed-batch culture, the NaCl concentration was increased to 7.5% (w/v) to trigger ectoine synthesis, while nitrogen and phosphorus sources were fed only during the first 3 h and then stopped to favor PHB accumulation. The process resulted in PHB yield of 68.5 wt.% of cell dry weight and volumetric productivity of about 1 g L⁻¹ h⁻¹ and ectoine concentration, content, and volumetric productivity of 4.3 g L⁻¹, 7.2 wt.%, and 2.8 g L⁻¹ day⁻¹, respectively. At salt concentration of 12.5% (w/v) during the second cultivation, the ectoine content was increased to 17 wt.% and productivity to 3.4 g L⁻¹ day⁻¹.     

Biological breakdown of denitrifying sulfide removal process in high-rate expanded granular bed reactor

This work conducted a denitrifying sulfide removal (DSR) test in an expanded granular sludge bed (EGSB) reactor at sustainable loadings of 6.09 kg m⁻³ day⁻¹ for sulfide, 3.11 kg m⁻³ day⁻¹ for nitrate–nitrogen, and 3.27 kg m⁻¹ day⁻¹ for acetate–carbon with >93% efficiency, which is significantly higher than those reported in literature. Strains Pseudomonas sp., Nitrincola sp., and Azoarcus sp. very likely yield heterotrophs. Strains Thermothrix sp. and Sulfurovum sp. are the autotrophs required for the proposed high-rate EGSB-DSR system. The EGSB-DSR reactor experienced two biological breakdowns, one at loadings of 4.87, 2.13, and 1.82 kg m⁻³ day⁻¹; reactor function was restored by increasing nitrate and acetate loadings. Another breakdown occurred at loadings of up to 8.00, 4.08, and 4.50 kg m⁻¹ day⁻¹; the heterotrophic denitrification pathway declined faster than the autotrophic pathway. The mechanism of DSR breakdown is as follows. High sulfide concentration inhibits heterotrophic denitrifiers, and the system therefore accumulates nitrite. Autotrophic denitrifiers are then inhibited by the accumulated nitrite, thereby leading to breakdown of the DSR process.     

Physiological differences in the growth of <Emphasis Type="Italic">Sargassum horneri</Emphasis> between the germling and adult stages

The effects of temperature, irradiance, and daylength on Sargassum horneri growth were examined at the germling and adult stages to discern their physiological differences. Temperature–irradiance (10, 15, 20, 25, 30°C × 20, 40, 80 μmol photons m⁻²s⁻¹) and daylength (8, 12, 16, 24 h) experiments were carried out. The germlings and blades of S. horneri grew over a wide range of temperatures (10–25°C), irradiances (20–80 μmol photons m⁻²s⁻¹), and daylengths (8–24 h). At the optimal growth conditions, the relative growth rates (RGR) of the germlings were 21% day⁻¹ (25°C, 20 μmol photons m⁻²s⁻¹) and 13% day⁻¹ (8 h daylength). In contrast, the RGRs of the blade weights were 4% day⁻¹ (15°C, 20 μmol photons m⁻²s⁻¹) and 5% day⁻¹ (12 h daylength). Negative growth rates were found at 20 μmol photons m⁻²s⁻¹ of 20°C and 25°C treatments after 12 days. This phenomenon coincides with the necrosis of S. horneri blades in field populations. In conclusion, we found physiological differences between S. horneri germlings and adults with respect to daylength and temperature optima. The growth of S. horneri germlings could be enhanced at 25°C, 20 μmol photons m⁻²s⁻¹, and 8 h daylength for construction of Sargassum beds and restoration of barren areas.     

Kinetics of CO conversion into H<Subscript>2</Subscript> by <Emphasis Type="Italic">Carboxydothermus hydrogenoformans</Emphasis>

The objective of this study was to improve the biological water–gas shift reaction for producing hydrogen (H₂) by conversion of carbon monoxide (CO) using an anaerobic thermophilic pure strain, Carboxydothermus hydrogenoformans. Specific hydrogen production rates and yields were investigated at initial biomass densities varying from 5 to 20 mg volatile suspended solid (VSS) L⁻¹. Results showed that the gas–liquid mass transfer limits the CO conversion rate at high biomass concentrations. At 100-rpm agitation and at CO partial pressure of 1 atm, the optimal substrate/biomass ratio must exceed 5 mol CO g⁻¹ biomass VSS in order to avoid gas–liquid substrate transfer limitation. An average H₂ yield of 94 ± 3% and a specific hydrogen production rate of ca. 3 mol g⁻¹ VSS day⁻¹ were obtained at initial biomass densities between 5 and 8 mg VSS⁻¹. In addition, CO bioconversion kinetics was assessed at CO partial pressure from 0.16 to 2 atm, corresponding to a dissolved CO concentration at 70°C from 0.09 to 1.1 mM. Specific bioactivity was maximal at 3.5 mol CO g⁻¹ VSS day⁻¹ for a dissolved CO concentration of 0.55 mM in the culture. This optimal concentration is higher than with most other hydrogenogenic carboxydotrophic species.