Growth responses of the perennial legume Sesbania sesban to NH4 and NO3 nutrition and effects on root nodulation

Preference for NH₄⁺ or NO₃⁻ nutrition by the perennial legume Sesbania sesban (L.) Merr. was assessed by supplying plants with NH₄⁺ and NO₃⁻ alone or mixed at equal concentrations (0.5 mM) in hydroponic culture. In addition, growth responses of S. sesban to NH₄⁺ and NO₃⁻ nutrition and the effects on root nodulation and nutrient and mineral composition of the plant tissues were evaluated in a hydroponic setup at a range of external concentration of NH₄⁺ and NO₃⁻ (0, 0.1, 0.2, 0.5, 2 and 5 mM). Seedlings of S. sesban grew equally well when supplied with either NH₄⁺ or NO₃⁻ alone or mixed and had high relative growth rates (RGRs) ranging between 0.19 and 0.21 d⁻¹. When larger plants of S. sesban were supplied with NH₄⁺ or NO₃⁻ alone, the RGRs and shoot elongation rates were not affected by the external concentration of inorganic N. At external N concentrations up to 0.5 mM nodulation occurred and contributed to the N nutrition through fixation of gaseous N₂ from the atmosphere. For both NH₄⁺ and NO₃⁻-fed plants the N concentration in the plant tissues, particularly water-extractable NO₃⁻, increased at high supply concentrations, and concentrations of mineral cations generally decreased. It is concluded that S. sesban can grow without an external inorganic N supply by fixing atmospheric N₂ gas via root nodules. Also, S. sesban grows well on both NH₄⁺ and NO₃⁻ as the external N source and the plant can tolerate relatively high concentrations of NH₄⁺. This wide ecological amplitude concerning N nutrition makes S. sesban very useful as a N₂-fixing fallow crop in N deficient areas and also a candidate species for use in constructed wetland systems for the treatment of NH₄⁺ rich waters.     

Nitrogen utilization by the raphidophyte Heterosigma akashiwo: Growth and uptake kinetics in laboratory cultures

The nitrogen uptake and growth capabilities of the potentially harmful, raphidophycean flagellate Heterosigma akashiwo (Hada) Sournia were examined in unialgal batch cultures (strain CCMP 1912). Growth rates as a function of three nitrogen substrates (ammonium, nitrate and urea) were determined at saturating and sub-saturating photosynthetic photon flux densities (PPFDs). At saturating PPFD (110 μE m⁻² s⁻¹), the growth rate of H. akashiwo was slightly greater for cells grown on NH₄⁺ (0.89 d⁻¹) compared to cells grown on NO₃⁻ or urea, which had identical growth rates (0.82 d⁻¹). At sub-saturating PPFD (40 μE m⁻² s⁻¹), both urea- and NH₄⁺-grown cells grew faster than NO₃⁻-grown cells (0.61, 0.57 and 0.46 d⁻¹, respectively). The N uptake kinetic parameters were investigated using exponentially growing batch cultures of H. akashiwo and the ¹⁵N-tracer technique. Maximum specific uptake rates (V_max) for unialgal cultures grown at 15 °C and saturating PPFD (110 μE m⁻² s⁻¹) were 28.0, 18.0 and 2.89 × 10⁻³ h⁻¹ for NH₄⁺, NO₃⁻ and urea, respectively. The traditional measure of nutrient affinity—the half saturation constants (K_s) were similar for NH₄⁺ and NO₃⁻ (1.44 and 1.47 μg-at N L⁻¹), but substantially lower for urea (0.42 μg-at N L⁻¹). Whereas the α parameter (α = V_max/K_s), which is considered a more robust indicator for substrate affinity when substrate concentrations are low (<K_s), were 19.4, 12.2 and 6.88 × 10⁻³ h⁻¹/(μg-at N L⁻¹) for NH₄⁺, NO₃⁻ and urea, respectively. These laboratory results demonstrate that at both saturating and sub-saturating N concentrations, N uptake preference follows the order: NH₄⁺ > NO₃⁻ > urea, and suggests that natural blooms of H. akashiwo may be initiated or maintained by any of the three nitrogen substrates examined.     

Influence of bed media characteristics on ammonia and nitrate removal in shallow horizontal subsurface flow constructed wetlands

Two bed media were tested (gravel and Filtralite) in shallow horizontal subsurface flow (HSSF) constructed wetlands in order to evaluate the removal of ammonia and nitrate for different types of wastewater (acetate-based and domestic wastewater) and different COD/N ratios. The use of Filtralite allowed both higher mass removal rates (1.1 g NH₄–N m⁻² d⁻¹ and 3 g NO₃–N m⁻² d⁻¹) and removal efficiencies (>62% for ammonia, 90–100% for nitrate), in less than 2 weeks, when compared to the ones observed with gravel. The COD/N ratio seems to have no significant influence on nitrate removal and the removal of both ammonia and nitrate seems to have involved not only the conventional pathways of nitrification–denitrification. The nitrogen loading rate of both ammonia (0.8–2.4 g NH₄–N m⁻² d⁻¹) and nitrate (0.6–3.2 g NO₃–N m⁻² d⁻¹) seem to have influenced the respective removal rates.     

Nitrogen Dynamics in the Steeply Stratified,Temperate Lake Verevi,Estonia

The dynamics of different nitrogen compounds and nitrification in diverse habitats of a stratified Lake Verevi (Estonia) was investigated in 2000–2001. Also planktonic N₂-fixation (N₂fix) was measured in August of the observed years. The nitrogen that accumulated in the hypolimnion was trapped in the non-mixed layer during most of the vegetation period causing a concentration of an order of magnitude higher than in the epilimnion. The ammonium level remained low in the epilimnion (maximum 577 mgN m⁻³, average 115 mgN m⁻³) in spite of high concentrations in the hypolimnion (maximum 12223 mgN m⁻³, average 4807 mgN m⁻³). The concentrations of NO₂⁻ and NO₃⁻ remained on a low level both in the epilimnion (average 0.94 and 9.09 mgN m⁻³, respectively) and hypolimnion (average 0.47 and 5.05 mgN m⁻³, respectively). N₂fix and nitrification ranged from 0.30 to 2.80 mgN m⁻³ day⁻¹ and 6.0 to 107 mgN m⁻³ day⁻¹, respectively; the most intensive processes occurred in 07.08.00 at depths of 2 and 5 m, accordingly. The role of N₂fix in the total nitrogen budget of Lake Verevi (in August 2000 and 2001) was negligible while episodically in the nitrogen-depleted epilimnion the N₂fix could substantially contribute to the pool of mineral nitrogen. Nitrification was unable to influence nitrogen dynamics in the epilimnion while some temporary coupling with ammonium dynamics in the hypolimnion was documented.     

Modelling of parameters for optimization of maturity in composting trimming residues

In this paper, changes in physico-chemical parameters during trimmings residue composting (cation exchange capacity, germination index, self-heated, NH₄/NO₃ ratio and C_FA/C_HA ratio) in relation to environmental composting parameters (time, aeration, moisture and particle size) of the composting process were studied. A central composite experimental design was used to obtain the polynomial model for each dependent variable. Results of the modelling showed that among the studied range, moisture was the highest influenced parameter in maturity evaluation, with respect to aeration and particle size. An exception was found for CEC evolution. In this parameter, the highest influence was found for particle size. Moreover, a product with acceptable chemical properties entails operating at medium moisture content (55%) and medium-to-high particle size (3–5 cm). Moderate to low aeration (0.2 m³ air kg⁻¹ d⁻¹) would be the best compromise to composting this residue, due to the scarce statistical influence of this independent variable.     

Annual cycle of CO2 exchange over a reed (Phragmites australis) wetland in Northeast China

Net ecosystem exchange of CO₂ (NEE) was measured during 2005 using the eddy covariance (EC) technique over a reed (Phragmites australis (Cav.) Trin. ex Steud.) wetland in Northeast China (121°54′E, 41°08′N). Diurnal NEE patterns varied markedly among months. Outside the growing season, NEE lacked a diurnal pattern and it fluctuated above zero with an average value of 0.07 mg CO₂ m⁻² s⁻¹ resulting from soil microbial activity. During the growing season, NEE showed a distinct V-like diel course, and the mean daily NEE was −7.48 ± 2.74 g CO₂ m⁻² day⁻¹, ranging from −13.58 g CO₂ m⁻² day⁻¹ (July) to −0.10 g CO₂ m⁻² day⁻¹ (October). An annual cycle was also apparent, with CO₂ uptake increasing rapidly in May, peaking in July, and decreasing from August. Monthly cumulative NEE ranged from −115 ± 24 g C m⁻² month⁻¹ (the reed wetland was a CO₂ sink) in July to 75 ± 16 g C m⁻² month⁻¹ (CO₂ source) in November. The annual CO₂ balance suggests a net uptake of −65 ± 14 g C m⁻² year⁻¹, mainly due to the gains in June and July. Cumulative CO₂ emission during the non-growing season was 327 g C m⁻², much greater than the absolute value of the annual CO₂ balance, which proves the importance of the wintertime CO₂ efflux at the study site. The ratio of ecosystem respiration (R_eco) to gross primary productivity (GPP) for this reed ecosystem was 0.95, indicating that 95% of plant assimilation was consumed by the reed plant or supported the activities of heterotrophs in the soil. Daytime NEE values during the growing season were closely related to photosynthetically active radiation (PAR) (r² > 0.63, p < 0.01). Both maximum ecosystem photosynthesis rate (A_max) and apparent quantum yield (α) were season-dependent, and reached their peak values in July (1.28 ± 0.11 mg CO₂ m⁻² s⁻¹, 0.098 ± 0.027 μmol CO₂ μmol⁻¹ photon, respectively), corresponding to the observed maximum NEE in July. Ecosystem respiration (R_eco) relied on temperature and soil water content, and the mean value of Q₁₀ was about 2.4 with monthly variation ranging from 1.8 to 4.1 during 2005. Annual methane emission from this reed ecosystem was estimated to be about 3 g C m⁻² year⁻¹, and about 5% of the net carbon fixed by the reed wetland was released to the atmosphere as CH₄.     

Application of response-surface methodology to evaluate the optimum medium components for the enhanced production of lichenysin by Bacillus licheniformis R2

Biosurfactants have gained attention because they exhibit some advantages such as biodegradability, low toxicity, ecological acceptability and ability to be produced from renewable and cheaper substrates. They are widely used for environmental applications for bioremediation and also in biomedical field. However, the high cost of production is the limiting factor for widespread industrial applications. Thus, optimization of the growth medium for biosurfactant-lichenysin production by Bacillus licheniformis R2 was carried out using response-surface methodology. A preliminary screening phase based on a two-level fractional factorial design led to the identification of NH₄NO₃, glucose, Na₂HPO₄ and MnSO₄·4H₂O concentrations as the most significant variables affecting the fermentation process. The 2⁴ full-factorial central composite design was then applied to further optimize the biosurfactant production. The optimal levels of the aforementioned variables were (g/l): NH₄NO₃, 1.0; glucose, 34.0; KH₂PO₄, 6.0; Na₂HPO₄, 2.7; MgSO₄·7H₂O, 0.1; CaCl₂, 1.2 × 10⁻³; FeSO₄·7H₂O, 1.65 × 10⁻³; MnSO₄·4H₂O, 1.5 × 10⁻³ and Na–EDTA, 2.2 × 10⁻³. With the optimization procedure, the relative lichenysin yield expressed as the critical micelle dilution (CMD) was fourfold higher than that obtained in the non-optimized reference medium.     

Utilization of the cyanobacteria Anabaena sp. ATCC 33047 in CO2 removal processes

In this paper the utilization of the cyanobacteria Anabaena sp. in carbon dioxide removal processes is evaluated. For this, continuous cultures of this strain were performed at different dilution rates; alternatives for the recovery of the organic matter produced being also studied. A maximum CO₂ fixation rate of 1.45 g CO₂ L⁻¹ day⁻¹ was measured experimentally, but it can be increased up to 3.0 g CO₂ L⁻¹ day⁻¹ outdoors. The CO₂ is mainly transformed into exopolysaccharides, biomass representing one third of the total organic matter produced. Organic matter can be recovered by sedimentation with efficiencies higher than 90%, the velocity of sedimentation being 2 · 10⁻⁴ s⁻¹. The major compounds were carbohydrates and proteins with productivities of 0.70 and 0.12 g L⁻¹ day⁻¹, respectively. The behaviour of the cultures of Anabaena sp. has been modelized, also the characteristics parameters requested to design separation units being reported. Finally, to valorizate the organic matter as biofertilizers and biofuels is proposed.     

Effects of NaCl salinity on growth, morphology, photosynthesis and proline accumulation of Salvinia natans

The effects of NaCl salinity on growth, morphology and photosynthesis of Salvinia natans (L.) All. were investigated by growing plants in a growth chamber at NaCl concentrations of 0, 50, 100 and 150 mM. The relative growth rates were high (ca. 0.3 d⁻¹) at salinities up to 50 mM and decreased to less than 0.2 d⁻¹ at higher salinities, but plants produced smaller and thicker leaves and had shorter stems and roots, probably imposed by the osmotic stress and lowered turgor pressure restricting cell expansion. Na⁺ concentrations in the plant tissue only increased three-fold, but uptake of K⁺ was reduced, resulting in very high Na⁺/K⁺ ratios at high salinities, indicating that S. natans lacks mechanisms to maintain ionic homeostasis in the cells. The contents of proline in the plant tissue increased at high salinity, but concentrations were very low (<0.1 μmol g⁻¹ FW), indicating a limited capacity of S. natans to synthesize proline as a compatible compound. The potential photochemical efficiency of PSII (F_v/F_m) of S. natans remained unchanged at 50 mM NaCl but was reduced at higher salinities, and the photosynthetic capacity (ETR_max) was significantly reduced at 50 mM NaCl and higher. It is concluded that S. natans is a salt-sensitive species lacking physiological measures to cope with exposure to high NaCl salinity. At low salinities salts are taken up and accumulate in old leaves, and high growth rates are maintained because new leaves are produced at a higher rate than for plants not exposed to salt.     

Microcosm assays and Taguchi experimental design for treatment of oil sludge containing high concentration of hydrocarbons

Microcosm assays and Taguchi experimental design was used to assess the biodegradation of an oil sludge produced by a gas processing unit. The study showed that the biodegradation of the sludge sample is feasible despite the high level of pollutants and complexity involved in the sludge. The physicochemical and microbiological characterization of the sludge revealed a high concentration of hydrocarbons (334,766 ± 7001 mg kg⁻¹ dry matter, d.m.) containing a variety of compounds between 6 and 73 carbon atoms in their structure, whereas the concentration of Fe was 60,000 mg kg⁻¹ d.m. and 26,800 mg kg⁻¹ d.m. of sulfide. A Taguchi L₉ experimental design comprising 4 variables and 3 levels moisture, nitrogen source, surfactant concentration and oxidant agent was performed, proving that moisture and nitrogen source are the major variables that affect CO₂ production and total petroleum hydrocarbons (TPH) degradation. The best experimental treatment yielded a TPH removal of 56,092 mg kg⁻¹ d.m. The treatment was carried out under the following conditions: 70% moisture, no oxidant agent, 0.5% of surfactant and NH₄Cl as nitrogen source.     

Effect of renoguanylin on hydrogen/bicarbonate ion transport in rat renal tubules

Renoguanylin (REN) is a recently described member of the guanylin family, which was first isolated from eels and is expressed in intestinal and specially kidney tissues. In the present work we evaluate the effects of REN on the mechanisms of hydrogen transport in rat renal tubules by the stationary microperfusion method. We evaluated the effect of 1 μM and 10 μM of renoguanylin (REN) on the reabsorption of bicarbonate in proximal and distal segments and found that there was a significant reduction in bicarbonate reabsorption. In proximal segments, REN promoted a significant effect at both 1 and 10 μM concentrations. Comparing control and REN concentration of 1 μM, JHCO₃⁻, nmol cm^− 2 s^− 1 − 1,76 ± 0,11_control × 1,29 ± 0,08_{REN 10 μM}; P < 0.05, was obtained. In distal segments the effect of both concentrations of REN was also effective, being significant e.g. at a concentration of 1 μM (JHCO₃⁻, nmol cm^− 2 s⁻ 1 − 0.80 ± 0.07_control × 0.60 ± 0.06_{REN 1 μM}; P < 0.05), although at a lower level than in the proximal tubule. Our results suggest that the action of REN on hydrogen transport involves the inhibition of Na⁺/H⁺exchanger and H⁺-ATPase in the luminal membrane of the perfused tubules by a PKG dependent pathway.     

Electricity from landfill leachate using microbial fuel cells: Comparison with a biological aerated filter

Four experimental columns were employed in this study to investigate their performance under wastewater treatment conditions. One column was set-up as a biological aerated filter and the remaining three were set-up as microbial fuel cells (MFCs), two of which were connected to an external load whereas the third was left open circuit. The performance of the columns under several flow rates and leachate strengths was studied in terms of BOD₅ removal efficiencies and electricity generation, when a fixed resistive load was connected. Results obtained demonstrated that it is possible to generate electricity and simultaneously treat landfill leachate in MFC columns. Energy generation in MFC columns improved with increasing flow rates from 24 to 192 mL/h, while BOD₅ removal efficiency levels reached a maximum at 48 mL/h and dropped to relatively low values at higher flow rates. The maximum removal efficiencies were obtained at a loading rate of 0.81 kg BOD₅/m³ d for columns C1, C2 and C4 and 1.81 kg BOD₅/m³ d for column C3. Electrical output levels and BOD₅ concentrations at the MFC columns showed a linear relationship, which allows the system to be used as a BOD₅ sensor. Part of the BOD removal was not associated with power generation and was attributed to the presence of alternative end terminal electron acceptors and volatilisation. The MFC columns could reach the same or even higher removal efficiencies than those from the biological aerated filter with the advantage of producing energy and saving cost of aeration. To the best of the authors’ knowledge, this is the first study that compares the MFC technology with other conventional treatment systems for removing pollutants from wastewater.     

Kinetics of ammonium, nitrate and phosphorus uptake by Canna indica and Schoenoplectus validus

Canna indica L. is an upright perennial rhizomatous herb, and Schoenoplectus validus (Vahl) A. Löve and D. Löve is a tall, perennial, herbaceous sedge. The nutrient uptake kinetics of C. indica and S. validus were investigated using the modified depletion method after plants were grown for 4 weeks in simulated secondary-treated wastewater. The maximum uptake rate (I_max) and Michaelis–Menten constant (K_m) were estimated by iterative curve fitting. The I_max for NH₄N (623 μmol g⁻¹ dry root weight h⁻¹) was significantly higher than that for NO₃N (338 μmol g⁻¹ dry root weight h⁻¹) in S. validus. In contrast, no difference was observed in C. indica. The I_max values for NO₃N and NH₄N were higher in S. validus than in C. indica. A significantly lower K_m was detected for NO₃N uptake in C. indica (385 μmol L⁻¹) compared to that in S. validus (1908 μmol L⁻¹). The I_max for PO₄P did not differ between the plant species. The K_m for PO₄P was significantly higher in C. indica (157 μmol L⁻¹) than in S. validus (60 μmol L⁻¹). In conclusion, we found that S. validus preferred NH₄N over NO₃N, had greater capacity for N uptake and higher affinity for PO₄P, but C. indica had greater affinity for NO₃N. Nutrient uptake capacity is likely related to habitat preference, and is influenced by the structure of roots and rhizomes.     

Ecophysiological responses of the euhalophyte Suaeda salsa to the interactive effects of salinity and nitrate availability

Effects of salinity and nitrate nitrogen (NO₃⁻-N) on ion accumulation and chlorophyll fluorescence were monitored for two populations of Suaeda salsa grown from seeds in a greenhouse experiment. One population inhabits the intertidal zone and the other occurs on inland saline soils. Ion contents in soils and in leaves of the two populations were also investigated in field. In the greenhouse, seedlings were exposed to a NaCl concentration of 0.6 and 35.1 ppt, with 0.1 or 5 mM NO₃⁻-N treatments for 20 days. The contents of Na⁺ and Cl⁻ were higher, but NO₃⁻ was lower in soils of the intertidal zone than at the inland site. In the field, ion concentrations and the estimated contribution of these ions to osmotic potential in leaves showed no difference between the two populations, except that the estimated contribution of Na⁺ to osmotic potential in leaves of the intertidal population was lower than that in the inland population. In the greenhouse, in contrast, the concentration of Cl⁻ was lower, but NO₃⁻ concentration and the estimated contribution of NO₃⁻ to osmotic potential were higher, in the leaves of plants from the intertidal zone. Salinity had no effect on the maximal efficiency of PSII photochemistry (Fv/Fm) and the actual PSII efficiency (ΦPSII). The results indicated that S. salsa from the intertidal zone was better able to regulate Cl⁻ to a lower level, and accumulate NO₃⁻ even with low soil NO₃⁻ concentrations. Tolerance of the PSII machinery to high salinity stress may be an important characteristic for the studied species supporting growth in highly saline environments.     

Gas exchange of Ginkgo biloba leaves at different CO2 concentration levels

One and a half year-old Ginkgo saplings were grown for 2 years in 7 litre pots with medium fertile soil at ambient air CO₂ concentration and at 700 μmol mol⁻¹ CO₂ in temperature and humidity-controlled cabinets standing in the field. In the middle of the 2nd season of CO₂ enrichment, CO₂ exchange and transpiration in response to CO₂ concentration was measured with a mini-cuvette system. In addition, the same measurements were conducted in the crown of one 60-year-old tree in the field. Number of leaves/tree was enhanced by elevated CO₂ and specific leaf area decreased significantly.CO₂ compensation points were reached at 75–84 μmol mol⁻¹ CO₂. Gas exchange of Ginkgo saplings reacted more intensively upon CO₂ than those of the adult Ginkgo. On an average, stomatal conductance decreased by 30% as CO₂ concentration increased from 30 to 1000 μmol mol⁻¹ CO₂. Water use efficiency of net photosynthesis was positively correlated with CO₂ concentration levels. Saturation of net photosynthesis and lowest level of stomatal conductance was reached by the leaves of Ginkgo saplings at >1000 μmol mol⁻¹ CO₂. Acclimation of leaf net CO₂ assimilation to the elevated CO₂ concentration at growth occurred after 2 years of exposure. Maximum of net CO₂ assimilation was 56% higher at ambient air CO₂ concentration than at 700 μmol mol⁻¹ CO₂.     

Treatment of an artificial sulphide containing wastewater in subsurface horizontal flow laboratory-scale constructed wetlands

In general, treatment wetlands seem to be a potential method of tackling the sulphide problem of post-treatment of anaerobic digester effluents.Because of insufficient practical experience and lack of knowledge of sulphide removal, sulphur transformation was investigated, particularly in horizontal subsurface flow constructed wetlands (depth of 35 cm) under laboratory-scale conditions with artificial wastewater.The plants affected a clear stimulation of the sulphide and ammonia removal rates. Sulphide concentration in the range of 1.5–2.0 mg l⁻¹ was tolerated by the plants and completely removed in the planted model wetlands; sulphide concentration of >2.0 mg l⁻¹ caused instabilities in sulphide and nitrogen removal. Area-specific sulphide removal rates of up to 94 mg sulphide m⁻² d⁻¹ were achieved in the planted beds at hydraulic retention times of 2.5 d. Sulphate affected the sulphide removal. While in the unplanted control bed an almost stable removal in the range of 150–300 mg N m⁻² d⁻¹ was observed variations of hydraulic retention time, sulphide and sulphate concentrations influenced the ammonia removal rate within the planted beds in a broader range (600–1400 mg N m⁻² d⁻¹).These results showed that nitrification, sulphide oxidation, denitrification and sulphate reduction can occur simultaneously in the rhizosphere of treatment wetlands caused by dynamic redox gradients (aerobic–anaerobic) conditions.     

Functional Expression of Dipeptidyl Peptidase I (Cathepsin C) of the Oriental Blood Fluke Schistosoma japonicum in Trichoplusia ni Insect Cells

Proenzyme dipeptidyl peptidase I (DPP I) of Schistosoma japonicum was expressed in a baculovirus expression system utilizing Trichoplusia ni BTI-5B1-4 (High Five) strain host insect cells. The recombinant enzyme was purified from cell culture supernatants by affinity chromatography on nickel–nitriloacetic acid resin, exploiting a polyhistidine tag fused to the COOH-terminus of the recombinant protease. The purified recombinant enzyme resolved in reducing SDS–PAGE gels as three forms, of 55, 39, and 38 kDa, all of which were reactive with antiserum raised against bacterially expressed S. japonicum DPP I. NH₂-terminal sequence analysis of the 55-kDa polypeptide revealed that it corresponded to residues −180 to −175, NH₂-SRXKXK, of the proregion peptide of S. japonicum DPP I. The 39- and 38-kDa polypeptides shared the NH₂-terminal sequence, LDXNQLY, corresponding to residues −73 to −67 of the proregion peptide and thus were generated by removal of 126 residues from the NH₂-terminus of the proenzyme. Following activation for 24 h at pH 7.0, 37°C under reducing conditions, the recombinant enzyme exhibited exopeptidase activity against synthetic peptidyl substrates diagnostic of DPP I. Specificity constants (k_cat/K_m) for the recombinant protease for the substrates H-Gly-Arg-NHMec and H-Gly-Phe-NHMec were found to be 14.4 and 10.7 mM⁻1 s⁻¹, respectively, at pH 7.0. Approximately 1 mg of affinity-purified schistosome DPP I was obtained per liter of insect cell culture supernatant, representing 2 × 10⁹ High Five cells.     

Behaviour of water bound in bone marrow cells affected by organic solvents of different polarity

The behaviour of intracellular water affected by organic solvents of different polarity in partially dehydrated marrow cells obtained from tubular bones of broiler chickens was studied using ¹H NMR spectroscopy at 210–290 K. The ¹H NMR spectra of intracellular water include two signals which can be assigned to strongly (SAW, chemical shift of the proton resonance δ_H = 4–5 ppm) and weakly (WAW, δ_H = 1.2–1.7 ppm) associated waters which can be also divided into weakly (WBW, frozen at 250 < T < 273 K and changes in the Gibbs free energy ΔG > −0.8 kJ/mol) and strongly (SBW, unfrozen at T < 250 K, ΔG < −0.8 kJ/mol) bound intracellular waters. Solvents of different polarity such as dimethylsulfoxide-d₆ (Me₂SO-d₆), acetonitrile-d₃, and chloroform-d differently affect structure, Gibbs free energy, and molecular mobility of intracellular water. A maximal fraction of SBW in WAW and a minimal fraction of SBW in SAW are observed on absorption of acetonitrile (0.8 g/g) by cells. The opposite results are on addition of Me₂SO (0.8 g/g) which strongly changes organisation of intracellular water and enhances the freezing point depression of SBW.     

Improvement of biodegradation in compact co-current biotrickling filter by high recycle liquid flow rate: Performance and biodegradation kinetics of ammonia removal

As a microbial-environmental-control-type deodorizing system, we have developed a compact biotrickling filter system for small-scale livestock farms. The performance of the compact co-current biotrickling filter operated at high recycle liquid flow rates was systematically examined. In particular, we studied improvements in the nitrification ability of the system due to the resultant enhancement of absorption and dissolution of NH₃ and absorption of O₂ with the high flow rates of recycle liquid flowing downward co-currently with gas flow. At the empty bed residence time of 50 s, almost complete removal of NH₃ was obtained with recycle liquid flow rates of 103 and 205 L m⁻³ day⁻¹ for 20 days while the inlet NH₃ concentration was increased from 200 to 500 ppm. With a recycle liquid flow rate of 411 L m⁻³ day⁻¹ the removal efficiency remained above 95% for 57 days while the inlet NH₃ concentration was increased from 200 to 700 ppm. The biodegradation kinetics for NH₃ removal was successfully analyzed using the Haldane substrate inhibition kinetics. The present data and kinetic analyses showed that the substrate inhibition was suppressed and the biodegradation of ammonia in the compact biotrickling filter could be improved by the high recycle liquid flow rate.     

Biodegradation of indole at high concentration by persolvent fermentation with the thermophilic fungus Sporotrichum thermophile

Indole and its derivatives form a class of toxic recalcitrant environmental pollulants. Sporotrichum thermophile was grown in a persolvent fermentation system containing a large amount of indole. The medium contained up to 20% by volume soybean oil and up to 2 g L⁻¹ indole. Most of the indole was partitioned in the organic solvent layer. When the organism was grown in the medium containing indole at 1 g L⁻¹, indole was totally consumed after 6 days. Under a fed–batch fermentation process where daily batches of indole (1 g L⁻¹) supplemented the microbial culture for 4 days, the biodegradation level was 3.0 g L⁻¹. These values make this process promising and worthy of further investigation for the microbial degradation of other toxic compounds.