Studies on Removing Sulfachloropyridazine from Groundwater with Microbial Bioreactors

Sulfachloropyridazine (SCP), an antibiotic used in aquaculture and in animal husbandry, is a common contaminant in surface and groundwaters. Two types of microbial reactors were evaluated as methods for removing SCP from flowing water. One type of reactor evaluated was a nitrogen-limiting biobarrier; the other a slow-sand-filter. Results showed that the soybean oil-fed, nitrogen-limiting biobarrier was not very effective at removing SCP from flowing water. When supplied with flowing water containing 2.4 mg l⁻¹ SCP the nitrogen-limiting biobarrier removed ~0.6 mg l⁻¹ SCP or about 28% of that present. SCP removal by the nitrogen-limiting biobarrier may not have been biological as abiotic removal was not ruled out. More efficient biological removal was obtained with the slow-sand-filter which reduced the SCP levels from 2.35 to 0.048 mg l⁻¹, a removal efficiency of ~98%. High levels of nitrate nitrogen, 50 mg l⁻¹ N, did not interfere with the removal processes of either reactor suggesting that SCP was not being degraded as a microbial nitrogen source.     

Microbial community of acetate utilizing denitrifiers in aerobic granules

Nitrite accumulates during biological denitrification processes when carbon sources are insufficient. Acetate, methanol, and ethanol were investigated as supplementary carbon sources in the nitrite denitrification process using biogranules. Without supplementary external electron donors (control), the biogranules degraded 200 mg l⁻¹ nitrite at a rate of 0.27 mg NO₂–N g⁻¹ VSS h⁻¹. Notably, 1,500 mg l⁻¹ acetate and 700 mg l⁻¹ methanol or ethanol enhanced denitrification rates for 200 mg l⁻¹ nitrite at 2.07, 1.20, and 1.60 mg NO₂–N g⁻¹ VSS h⁻¹, respectively; these rates were significantly higher than that of the control. The sodium dodecyl sulfate polyacrylamide gel electrophoresis of the nitrite reductase (NiR) enzyme identified three prominent bands with molecular weights of 37–41 kDa. A linear correlation existed between incremental denitrification rates and incremental activity of the NiR enzyme. The NiR enzyme activity was enhanced by the supplementary carbon sources, thereby increasing the nitrite denitrification rate. The capacity of supplementary carbon source on enhancing NiR enzyme activity follows: methanol > acetate > ethanol on molar basis or acetate > ethanol > methanol on an added weight basis.     

Biological Remediation of Groundwater Containing Both Nitrate and Atrazine

Due to its high usage, mobility, and recalcitrant nature, atrazine is a common groundwater contaminant. Moreover, groundwaters that are contaminated with atrazine often contain nitrate as well. Nitrate interferes with the biological degradation of atrazine and makes it more difficult to use in situ biological methods to remediate atrazine contaminated groundwater. To solve this problem we used two reactors in sequence as models of in situ biobarriers; the first was a vegetable-oil-based denitrifying biobarrier and the second an aerobic reactor that oxygenated the denitrifying reactor’s effluent. The reactors were inoculated with an atrazine-degrading microbial consortium and supplied with water containing 5 mg l⁻¹ nitrate–N and 3 mg l⁻¹ atrazine. Our hypothesis was that the denitrifying barrier would remove nitrate from the flowing water and that the downstream reaction would remove atrazine. Our hypothesis proved correct; the two reactor system removed 99.9% of the atrazine during the final 30 weeks of the study. The denitrifying barrier removed ~98% of the nitrate and ~30% of the atrazine while the aerobic reactor removed ~70% of the initial atrazine. The system continued to work when the amount of nitrate–N in the influent water was increased to 50 mg l⁻¹. A mercury poisoning study blocked the degradation of atrazine indicating that biological processes were involved. An in situ denitrifying barrier coupled with an air injection system or other oxygenation process might be used to remove both nitrate and atrazine from contaminated groundwater or to protect groundwater from an atrazine spill.     

Nitrate removal from drinking water using a membrane-fixed biofilm reactor

Biological treatment of drinking water is a cost-effective alternative to conventional physico/chemical processes. A new concept was tested to overcome the main disadvantage of biological denitrification, the intensive post-treatment process to remove microorganisms and remnant carbon source. The biological reaction zone and carbon supply were separated from the raw water stream by a nitrate-permeable membrane. Denitrification takes place in a biofilm, which is immobilized at the membrane. In a series of bench-scale runs, different types of membranes and reactor configurations were investigated. The best denitrification rates achieved were 1230 mg NO₃ ⁻-N m⁻² day⁻¹. In one run, raw water containing 100 mg NO₃ ⁻ l⁻¹ was completely freed from nitrate. The membrane and the attached biofilm also represent a barrier against the passage of the C source and nutrients into the raw water. At concentrations of 20 mg l⁻¹ ethanol and 15 mg l⁻¹ phosphate in the bioreactor no diffusion through the membrane into the treated water was observed. Without any post-treatment, the effluent met nearly all the relevant criteria for drinking water; only the colony count was slightly increased. Received: 18 December 1996 / Received last revision: 14 April 1997 / Accepted: 19 April 1997     

A sequencing batch reactor system for high-level biological nitrogen and phosphorus removal from abattoir wastewater

A sequencing batch reactor (SBR) system is demonstrated to biologically remove nitrogen, phosphorus and chemical oxygen demand (COD) to very low levels from abattoir wastewater. Each 6 h cycle contained three anoxic/anaerobic and aerobic sub-cycles with wastewater fed at the beginning of each anoxic/anaerobic period. The step-feed strategy was applied to avoid high-level build-up of nitrate or nitrite during nitrification, and therefore to facilitate the creation of anaerobic conditions required for biological phosphorus removal. A high degree removal of total phosphorus (>98%), total nitrogen (>97%) and total COD (>95%) was consistently and reliably achieved after a 3-month start-up period. The concentrations of total phosphate and inorganic nitrogen in the effluent were consistently lower than 0.2 mg P l⁻¹ and 8 mg N l⁻¹, respectively. Fluorescence in situ hybridization revealed that the sludge was enriched in Accumulibacter spp. (20–40%), a known polyphosphate accumulating organism, whereas the known glycogen accumulating organisms were almost absent. The SBR received two streams of abattoir wastewater, namely the effluent from a full-scale anaerobic pond (75%) and the effluent from a lab-scale high-rate pre-fermentor (25%), both receiving raw abattoir wastewater as feed. The pond effluent contained approximately 250 mg N l⁻¹ total nitrogen and 40 mg P l⁻¹ of total phosphorus, but relatively low levels of soluble COD (around 500 mg l⁻¹). The high-rate lab-scale pre-fermentor, operated at 37°C and with a sludge retention time of 1 day, proved to be a cheap and effective method for providing supplementary volatile fatty acids allowing for high-degree of biological nutrient removal from abattoir wastewater.     

A study on using fireclay as a biomass carrier in an activated sludge system

By adding a biomass carrier to an activated sludge system, the biomass concentration will increase, and subsequently the organic removal efficiency will be enhanced. In this study, the possibility of using excess sludge from ceramic and tile manufacturing plants as a biomass carrier was investigated. The aim of this study was to determine the effect of using fireclay as a biomass carrier on biomass concentration, organic removal and nitrification efficiency in an activated sludge system. Experiments were conducted by using a bench scale activated sludge system operating in batch and continuous modes. Artificial simulated wastewater was made by using recirculated water in a ceramic manufactutring plant. In the continuous mode, hydraulic detention time in the aeration reactor was 8 and 22 h. In the batch mode, aeration time was 8 and 16 h. Fireclay doses were 500, 1,400 and 2,250 mg l⁻¹, and were added to the reactors in each experiment separately. The reactor with added fireclay was called a Hybrid Biological Reactor (HBR). A reactor without added fireclay was used as a control. Efficiency parameters such as COD, MLVSS and nitrate were measured in the control and HBR reactors according to standard methods. The average concentration of biomass in the HBR reactor was greater than in the control reactor. The total biomass concentration in the HBR reactor (2.25 g l⁻¹ fireclay) in the continuous mode was 3,000 mg l⁻¹ and in the batch mode was 2,400 mg l⁻¹. The attached biomass concentration in the HBR reactor (2.25 g l⁻¹ fireclay) in the continuous mode was 1,500 mg l⁻¹ and in the batch mode was 980 mg l⁻¹. Efficiency for COD removal in the HBR and control reactor was 95 and 55%, respectively. In the HBR reactor, nitrification was enhanced, and the concentration of nitrate was increased by 80%. By increasing the fireclay dose, total and attached biomass was increased. By adding fireclay as a biomass carrier, the efficiency of an activated sludge system to treat wastewater from ceramic manufacturing plants was increased.     

Ethrel treatment enhanced isoflavonoids accumulation in cell suspension cultures of Pueraria tuberosa, a woody legume

The cell cultures of Pueraria tuberosa, a perennial leguminous lianas, were maintained in modified MS medium (KNO₃ 475 mg l⁻¹, thiamine 1 mg l⁻¹, biotin 1 mg l⁻¹, calcium pantothenate 1 mg l⁻¹) containing 0.1 mg l⁻¹ 2,4,5-trichloroacetic acid and 0.1 mg l⁻¹ kinetin. Isoflavonoids (puerarin, genistin, daidzein, genistein) accumulation in cell suspension cultures was increased by 14-fold to ~12 mg l⁻¹ after 48 h of adding 100 μM ethrel. Ethrel inhibitors (silver nitrate and silver thiosulfate) completely inhibited this effect in the presence of ethrel and isoflavonoids were not detected in the spent medium. The increase was dose dependent and can be explored to trigger high yield of isoflavonoids production.     

In vitro regeneration and morphogenesis studies in common bean

An efficient protocol for high frequency in vitro regeneration of multiple shoots and somatic embryos from the embryonic axis of common bean (Phaseolus vulgaris) was developed. Ten common bean cultivars representing a wide range of diversity among current commercial market classes were used for in vitro regeneration evaluation in our study. These cultivars were tested on 63 different media formulations consisting of combinations of cytokinins, namely benzyladenine (BA) and thidiazuron (TDZ) at concentration levels of 0.0, 1.0, 2.5, 5.0 and 10.0 mg l⁻¹ and auxin, namely naphthalene acetic acid (NAA) and indole-3-acetic acid (IAA) at concentration levels of 0.0, 0.05, 0.1 and 1.0 mg l⁻¹. P. vulgaris cv. Olathe pinto bean performed the best producing over 20 multiple shoots per explant while cv. Condor black bean was the poorest with nine multiple shoots per explant. The optimum media for regeneration of multiple shoots was 4.4 mg l⁻¹ Murashige and Skoog (MS) containing 2.5 mg l⁻¹ BA and 0.1 mg l⁻¹ IAA supplemented with 30 mg l⁻¹ silver nitrate. Adventitious shoots and somatic embryos were regenerated on 4.4 mg l⁻¹ MS medium containing 1 mg l⁻¹ TDZ and 0.05 mg l⁻¹ NAA supplemented with 30 mg l⁻¹ silver nitrate or activated charcoal. Efficient and effective rooting of plantlets was achieved by dipping the cut end base of in vitro regenerated shoots in 1.0 mg l⁻¹ indole-3-butyric acid (IBA) solution and culturing on media containing 4.4 mg l⁻¹ MS supplemented by 0.1 mg l⁻¹ IAA, NAA or IBA.     

Nitrate reduction by Citrobacter diversus under aerobic environment

A new aerobic denitrifier, Citrobacter diversus, was isolated from both nitrification and denitrification sludge. To monitor the variation in the concentration of nitrogen oxides, aerobic denitrification by C. diversus was carried out in a batch reactor. When the nitrate concentration was greater than 180 mg N l⁻¹, the nitrate reduction rate became stable. The effect of the C/N ratio on the denitrification activity was also investigated. The results showed that the optimum denitrification activity was obtained when the C/N ratio was 4–5. The range of the C/N ratio was higher than that for traditional anoxic denitrification. The effect of the dissolved oxygen concentration was further studied; and it was found that the range of dissolved oxygen concentrations, both for specific growth rates and for specific denitrification rates, was 2–6 mg⁻¹. From these results, it can be concluded that both the concentration of dissolved oxygen and the C/N ratio are key factors in the aerobic denitrification by C. diversus. Received: 23 November 1999 / Received revision: 4 February 2000 / Accepted: 13 February 2000     

Treatment of phenol in an anaerobic fluidized bed reactor (AFBR): continuous and batch regime

Results of this study describe the feasibility of anaerobic treatment of highly concentrated phenol synthetic wastewater using an anaerobic fluidized bed reactor (AFBR) in both continuous and batch modes. Wastewater with a maximum load of 2,100 mg C·l⁻¹ was prepared using phenol (maximum concentration of 1,600 mg C·l⁻¹) as substrate and a mixture of acetic, propionic and butyric acids (500 mg C·l⁻¹) as co-substrate. AFBR reached total organic carbon (TOC) and phenol removal efficiency over 95% treating the highest organic loading rate (OLR) containing phenol studied for this kind of reactor (5.03 g C·l⁻¹·d⁻¹). The phenol loading rate rise caused volumetric biogas rate increase up to 4.4 l·l⁻¹·d⁻¹ (average yield of 0.28 l CH₄·g⁻¹ COD_removed) as well as variation in the biogas composition; the CO₂ percentage increased while the CH₄ percentage decreased. Morphological examination of the bioparticles at 4.10 g C·l⁻¹·d⁻¹, revealed significant differences in the biofilm structure, microbial colonization and bacterial morphological type development. The five batch assays showed that phenol degradation may be favoured by the presence of volatile fatty acids (VFAs) (co-metabolism), whereas VFAs degradation may be inhibited by phenol. AFBR reached initial phenol degradation velocity of 0.25 mg C·l⁻¹·min⁻¹.     

Catalyzing denitrification of <Emphasis Type="Italic">Paracoccus versutus</Emphasis> by immobilized 1,5-dichloroanthraquinone

The accelerating effect of non-dissolved redox mediator (1,5-dichloroanthraquinone) on the biological denitrification was investigated in this paper using 1,5-dichloroanthraquinone immobilized by calcium alginate (CA) and a heterotrophic denitrification bacterium of Paracoccus versutus (GU111570). The results suggested that the denitrification rate was enhanced 2.1 fold by 25 mmol l⁻¹ 1,5-dichloroanthraquinone of this study, and a positive correlation was found for the denitrification rate and 1,5-dichloroanthraquinone concentrations from 0 to 25 mmol l⁻¹. According to the change characteristic of NO₃ ⁻ and NO₂ ⁻ during the denitrification process, the tentative accelerating mechanism of the denitrification by redox mediators was put forward, and redox mediator might play the role of reduced cofactors like NADH, N(A)DH and SDH, or the similar ubiquinol/ubiquinone (Q/QH₂) role during the denitrification process.     

Biotreatment of phenol-contaminated wastewater in a spiral packed-bed bioreactor

A spiral packed-bed bioreactor inoculated with microorganisms obtained from activated sludge was used to conduct a feasibility study for phenol removal. The reactor was operated continuously at various phenol loadings ranging from 53 to 201.4 g m⁻³ h⁻¹, and at different hydraulic retention times (HRT) in the range of 20–180 min to estimate the performance of the device. The results indicated that phenol removal efficiency ranging from 82.9 to 100% can be reached when the reactor is operated at an HRT of 1 h and a phenol loading of less than 111.9 g m⁻³ h⁻¹. At an influent phenol concentration of 201.4 g m⁻³, the removal efficiency increased from 18.6 to 76.9% with an increase in the HRT (20–120 min). For treatment of phenol in the reactor, the maximum biodegradation rate (V _m) was 1.82 mg l⁻¹ min⁻¹; the half-saturation constant (K _s), 34.95 mg l⁻¹.     

Effects of plant growth regulators and sucrose on post harvest physiology,membrane stability and vase life of cut spikes of gladiolus

Effects of post-harvest application of two plant growth regulators viz., gibberellic acid (GA₃) and benzyl adenine (BA) with sucrose in the vase solution on cell membrane stability and vase life of gladiolus were investigated. The vase solution treatment combinations of GA₃ and BA with sucrose significantly increased the membrane stability index and enhanced the vase life as compared to the sucrose alone treatments or the controls. Vase solution treatment of GA₃ (50 mg l⁻¹), followed by BA (50 mg l⁻¹) with sucrose (50 g l⁻¹) significantly increased solution uptake, fresh weight and dry weight of cut spikes. The same treatments also enhanced the concentration of reducing and non-reducing sugars in gladioli petals 4 days after treatment (DAT). Cut spikes in vase solution enriched with 50 mg l⁻¹ GA₃ + 50 g l⁻¹ sucrose showed higher antioxidative enzyme activities of superoxide dismutase (SOD) and glutathione reductase (GR), lower lipoxygenase (LOX) activity and lipid peroxidation (measured as TBARS). Petal membrane stability index was also highest in cut spikes 6 DAT with 50 mg l⁻¹ GA₃ + 50 g l⁻¹ sucrose vase solution. Treatment of gladiolus cut spikes with 50 mg l⁻¹ GA₃ + 50 g l⁻¹ sucrose vase solution showed two fold increase in vase life and improved flower quality with a higher number of open flower per spike at any one time. These results suggest that post-harvest application of GA₃ (50 mg l⁻¹) with sucrose (50 g l⁻¹) maintains higher spike fresh and dry weight, improves anti-oxidative defence, stabilizes membrane integrity leading to a delay in petal cell death.     

Simultaneous biological removal of sulfur, nitrogen and carbon using EGSB reactor

High-rate biological conversion of sulfide and nitrate in synthetic wastewater to, respectively, elemental sulfur (S⁰) and nitrogen-containing gas (such as N₂) was achieved in an expanded granular sludge bed (EGSB) reactor. A novel strategy was adopted to first cultivate mature granules using anaerobic sludge as seed sludge in sulfate-laden medium. The cultivated granules were then incubated in sulfide-laden medium to acclimate autotrophic denitrifiers. The incubated granules converted sulfide, nitrate, and acetate simultaneously in the same EGSB reactor to S⁰, N-containing gases and CO₂ at loading rates of 3.0 kg S m⁻³ d⁻¹, 1.45 kg N m⁻³ d⁻¹, and 2.77 kg Ac m⁻¹ d⁻¹, respectively, and was not inhibited by sulfide concentrations up to 800 mg l⁻¹. Effects of the C/N ratio on granule performance were identified. The granules cultivated in the sulfide-laden medium have Pseudomonas spp. and Azoarcus sp. presenting the heterotrophs and autotrophs that co-work in the high-rate EGSB-SDD (simultaneous desulfurization and denitrification) reactor.     

Fluoranthene influences endogenous abscisic acid level and primary photosynthetic processes in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.) plants in vitro

The influence of increasing concentrations (0.1, 1.0 and 5.0 mg l⁻¹) of fluoranthene (FLT) on growth, endogenous abscisic acid (ABA) level and primary photosynthetic processes in 21-day-old pea plants (Pisum sativum L.) in vitro was investigated. Murashige and Skoog’s (MS) medium, with or without FLT, was enriched with indole-3-acetic acid (IAA; 0.1 mg l⁻¹) or a combination of IAA (0.1 mg l⁻¹) plus N⁶-benzyladenine (BA; 0.1 mg l⁻¹). The level of endogenous ABA significantly increased with increasing FLT concentrations in the presence of both IAA and IAA plus BA. An increased level of endogenous ABA was observed in plants treated with IAA alone. The growth of shoot, callus and the content of photosynthetic pigments (chlorophyll a and b, carotenoids), in both IAA- and IAA plus BA-treated plants, were significantly stimulated by FLT at its lowest concentration (0.1 mg l⁻¹) assayed in this study. However, FLT at higher concentrations (1.0 and 5.0 mg l⁻¹) significantly inhibited all these parameters. Chlorophyll fluorescence imaging showed that FLT only at the highest concentration (5.0 mg l⁻¹) in the presence of IAA (0.1 mg l⁻¹) significantly increased F₀, but decreased F_V/F_M and Φ_II.     

Photoinhibition-induced reduction in photosynthesis is alleviated by abscisic acid,cytokinin and brassinosteroid in detached tomato leaves

Detached leaves of tomato (Lycopersicon esculentum Mill.) experienced photoinhibition associated with sharp reductions in net photosynthetic rate (Pn), quantum efficiency of PSII (Φ_PSII) and photochemical quenching (qP) even though they were exposed to mild light intensity (400 μmol m⁻² s⁻¹ PPFD) at 28°C. Photoinhibition and the reduction in Pn, Φ_PSII and qP, however, were significantly alleviated by 1 mg l⁻¹ ABA, 0.1 mg l⁻¹ N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) and 0.01 mg l⁻¹ 24-epibrassinolide (EBR). Higher concentrations, however, reduced the effects or even exacerbated the occurrence of photoinhibition. Superoxide dismutase and ascorbate peroxidase activity in leaves increased with the increases in ABA concentration within 1–100 mg l⁻¹, CPPU concentration within 0.1–10 mg l⁻¹ and EBR concentration within 0.01–1.0 mg l⁻¹. Catalase and guaiacol peroxidase activity also increased with the increase in EBR concentration but CPPU and ABA treatments at higher concentrations caused a decrease. Malondialdehyde (MDA) content decreased with the increase in CPPU concentration. ABA and EBR, however, decreased MDA concentration only at 1 and 0.01 mg l⁻¹, respectively. In conclusion, detached leaves had increased sensitivity to PSII photoinhibition. Photoinhibition-induced decrease in photosynthesis, however, was significantly alleviated by EBR, CPPU and ABA at a proper concentration.     

Enhanced regeneration of haploid plantlets from microspores of <Emphasis Type="Italic">Brassica napus</Emphasis> L. using bleomycin,PCIB, and phytohormones

The effects of three periods of incubation (10, 20 and 30 min) at different levels of bleomycin (0, 0.1, 0.2, 0.3, 0.4 and 0.5 μg ml⁻¹), as well as three periods of exposure (12, 24 and 48 h) to different levels of the anti-auxin p-chlorophenoxyisobutyric acid (PCIB), including 1, 2, 3, 4 and 5 mg l⁻¹, on microspore embryogenesis of rapeseed cv. ‘Amica’ were investigated. Microspore embryogenesis was significantly enhanced following 20 min treatment with 0.2 μg ml⁻¹ bleomycin compared with untreated cultures. Highest embryo yield (163 embryos Petri dish⁻¹) was observed with 24 h treatment of 4 mg l⁻¹ PCIB. The highest percentage of secondary embryogenesis was observed on B₅ medium containing 0.15 mg l⁻¹ of gibberellic acid (GA₃) and 0.2 mg l⁻¹ 6-benzyladenine (BA) in 4–6 mm microspore-derived embryos (MDEs). Most callus formed on B₅ medium containing 0.15 mg l⁻¹ GA₃, 0.1 mg l⁻¹ BA and 0.1 mg l⁻¹ indole-3-acetic acid (IAA) when 4–6 mm embryos were used. Regeneration was highest on B₅ medium containing 0.05 mg l⁻¹ GA₃ or 0.1 mg l⁻¹ BA and 0.2 mg l⁻¹ IAA with 2–4 mm embryos. Microspore embryogenesis and plant regeneration could be improved by both bleomycin and PCIB when the appropriate MDE length and phytohormone level were selected.     

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.     

Seasonal modulation of bacterioplankton community at a temperate eutrophic shallow lake

At the present time, there is still a lack of information about environmental parameters modulating variations on bacterial diversity in temperate lakes, particularly from Portugal. Fermentelos Lake (Central Portugal) is a shallow water body that sustains an important wetland area. The strong nutrient inputs from agriculture and industrial runoffs have led to its current eutrophic status. The present work aimed to understand which factors modulate the seasonal bacterioplankton diversity at this lake using 16S rRNA PCR-denaturing gradient gel electrophoresis (DGGE) and multivariate analysis. Environmental data demonstrated eutrophic features throughout all samples with nitrate concentrations reaching 12.0 mg N (NO₃ ⁻) l⁻¹ in March 2006, while the highest conductivity (609 μS cm⁻¹), soluble reactive phosphorus (0.37 mg l⁻¹), total suspended solids (87.2 mg l⁻¹) and chlorophyll a (286.6 μg l⁻¹) levels were recorded in August 2007. Over the past two decades there was a general increase in nitrate, pH and conductivity levels at this lake, suggesting the eutrophication process is still in progress. Multivariate analysis showed that summer versus winter DGGE patterns could be established for bacterial assemblages and were mainly defined by water temperature and chlorophyll a. Actinobacteria were dominant throughout the study period although a general preference for higher temperature, pH, total suspended solids, conductivity, soluble reactive phosphorus (SRP) and chlorophyll a levels was observed. The highest concentrations of nitrogen sources were related to Bacteroidetes and phototrophic eukaryote (cryptophycean) dominance. The expansion of Betaproteobacteria, Alphaproteobacteria and Cyanobacteria phylotypes was generally associated to high temperature, pH, conductivity and SRP values.     

Fluoranthene-induced production of ethylene and formation of lysigenous intercellular spaces in pea plants cultivated in vitro

The effect of increasing concentration of polycyclic aromatic hydrocarbon (PAH) fluoranthene (FLT; 0.1, 1 and 5 mg l⁻¹) on the growth, ethylene production and anatomy of stems of 21-day-old pea plants cultivated in vitro in MS medium, with or without FLT, enriched with 0.1 mg l⁻¹ indole-3-acetic acid (IAA) or with combination of 0.1 mg l⁻¹ IAA + 0.1 mg l⁻¹ N⁶-benzyladenine (BA) were investigated. The low concentration of 0.1 mg l⁻¹ FLT, in both IAA- and IAA + BA-treated plants, significantly stimulated the growth of pea callus, while higher concentrations 1 mg l⁻¹ and especially 5 mg l⁻¹ FLT significantly inhibited it. Pea shoots were significantly influenced only after application of 5 mg l⁻¹ FLT in IAA treatment. Significantly increased production of ethylene was found in IAA + BA treatments in all concentrations of FLT, whereas in IAA treatments in 1 and 5 mg l⁻¹ FLT. The lysigenous aerenchyma formation in the cortex of pea stems significantly increased in all FLT treatments and its highest proportion was found in plants exposed to 1 mg l⁻¹ FLT.