Hydrophobic response of the fungus <Emphasis Type="Italic">Rhinocladiella similis</Emphasis> in the biofiltration with volatile organic compounds with different polarity

Rhinocladiella similis biodegraded volatile organic compounds (VOCs) of different polarity in gas-phase biofilters. Elimination capacities, (EC) of 74 g_hexane m⁻³ h⁻¹, 230 g_ethanol m⁻³ h⁻¹, 85 g_toluene m⁻³ h⁻¹ and 30 g_phenol m⁻³ h⁻¹ were obtained. EC values correlated with the solubility of the VOCs. R. similis grown with n-hexane or ethanol in biofilters packed with Perlite showed that the surface hydrophobicity was higher with n-hexane than ethanol. The hydrophobin-like proteins extracted from the mycelium produced with n-hexane (15 kDa) were different from those in the ethanol biofilter (8.5 kDa and 7 kDa).     

Effects of Artemisinin Derivative on the Growth Metabolism of <Emphasis Type="Italic">Tetrahymena thermophila</Emphasis> BF5 Based on Expression of Thermokinetics

The toxic effects of artesunate and dihydroartemisinin on the growth metabolism of Tetrahymena thermophila BF5 were studied by microcalorimetry. The results showed that: (1) low concentrations of artesunate (≤1 mg L⁻¹) and dihydroartemisinin (≤ 2 mg L⁻¹) promoted the growth metabolism of T. thermophila BF5, whereas high concentrations of artesunate (1–60 mg L⁻¹) and dihydroartemisinin (2–60 mg L⁻¹) inhibited its growth; (2) the half inhibition concentrations IC₅₀ of artesunate and dihydroartemisinin were 17.5817 and 9.5089 mg L⁻¹, respectively. It was concluded that the inhibition of dihydroartemisinin was stronger than that of artesunate.     

BTEX catabolism interactions in a toluene-acclimatized biofilter

BTEX substrate interactions for a toluene-acclimatized biofilter consortium were investigated. Benzene, ethylbenzene, o-xylene, m-xylene and p-xylene removal efficiencies were determined at a loading rate of 18.07 g m⁻³h⁻¹ and retention times of 0.5–3.0 min. This was also repeated for toluene in a 1:1 (m/m) ratio mixture (toluene: benzene, ethylbenzene, or xylene ) with each of the other compounds individually to obtain a final total loading of 18.07 g m⁻³h⁻¹. The results obtained were modelled using Michaelis–Menten kinetics and an explicit finite difference scheme to generate v _max and K _m parameters. The v _max/K _m ratio (a measure of the catalytic efficiency, or biodegradation capacity, of the reactor) was used to quantify substrate interactions occurring within the biofilter reactor without the need for free-cell suspended and monoculture experimentation. Toluene was found to enhance the catalytic efficiency of the reactor for p-xylene, while catabolism of all the other compounds was inhibited competitively by the presence of toluene. The toluene-acclimatized biofilter was also able to degrade all of the other BTEX compounds, even in the absence of toluene. The catalytic efficiency of the reactor for compounds other than toluene was in the order: ethylbenzene>benzene>o-xylene>m-xylene>p-xylene. The catalytic efficiency for toluene was reduced by the presence of all other tested BTEX compounds, with the greatest inhibitory effect being caused by the presence of benzene, while o-xylene and p-xylene caused the least inhibitory effect. This work illustrated that substrate interactions can be determined directly from biofilter reactor results without the need for free-cell and monoculture experimentation. Received: 13 April 2000 / Received revision: 20 July 2000 / Accepted: 27 July 2000     

H2S degradation is reflected by both the activity and composition of the microbial community in a compost biofilter

In this study, 16S rRNA- and rDNA-based denaturing gradient gel electrophoresis (DGGE) were used to study the temporal and spatial evolution of the microbial communities in a compost biofilter removing H₂S and in a control biofilter without H₂S loading. During the first 81 days of the experiment, the H₂S removal efficiencies always exceeded 93% at loading rates between 4.1 and 30 g m⁻³ h⁻¹. Afterwards, the H₂S removal efficiency decreased to values between 44 and 71%. RNA-based DGGE analysis showed that H₂S loading to the biofilter increased the stability of the active microbial community but decreased the activity-based diversity and evenness. The most intense band in both the RNA- and DNA-based DGGE patterns of the H₂S-degrading biofilter represented the sulfur oxidizing bacterium Thiobacillus thioparus. This suggested that T. thioparus constituted a major part of the bacterial community and was an important primary degrader in the H₂S-degrading biofilter. The decreasing H₂S removal efficiencies near the end of the experiment were not accompanied by a substantial change of the DGGE patterns. Therefore, the decreased H₂S removal was probably not caused by a failing microbiology but rather by a decrease of the mass transfer of substrates after agglutination of the compost particles.     

Toluene degradation in a polyurethane biofilter at high loading

In the present study, toluene elimination in the polyurethane (PU) biofilter during long-term (145 day) operation was characterized, and assessed the effects of changing the inlet loading and space velocity (SV). A very high elimination capacity of 3.7 kg·m⁻³·h⁻¹ was obtained at an inlet loading of 4.0 kg·m^–3·h⁻¹ (inlet toluene concentration of 900 ppmv at a SV of 1,040 h⁻¹). Backwashing with irrigation and compressed air allowed maintenance of a pressure drop of < 80 mm H₂O·m⁻¹-filter at an SV of 830 h⁻¹ and an elimination efficiency of > 90% during the 145 day of operation. In conclusion, the PU biofilter can overcome the problems of clogging caused by excess biomass growth and of low treatment capacities of conventional biofilters.     

Production of hexanoic acid from d-galactitol by a newly isolated Clostridium sp. BS-1

In a study screening anaerobic microbes utilizing d-galactitol as a fermentable carbon source, four bacterial strains were isolated from an enrichment culture producing H₂, ethanol, butanol, acetic acid, butyric acid, and hexanoic acid. Among these isolates, strain BS-1 produced hexanoic acid as a major metabolic product of anaerobic fermentation with d-galactitol. Strain BS-1 belonged to the genus Clostridium based on phylogenetic analysis using 16S rRNA gene sequences, and the most closely related strain was Clostridium sporosphaeroides DSM 1294^T, with 94.4% 16S rRNA gene similarity. In batch cultures, Clostridium sp. BS-1 produced 550 ± 31 mL L⁻¹ of H₂, 0.36 ± 0.01 g L⁻¹ of acetic acid, 0.44 ± 0.01 g L⁻¹ of butyric acid, and 0.98 ± 0.03 g L⁻¹ of hexanoic acid in a 4-day cultivation. The production of hexanoic acid increased to 1.22 and 1.73 g L⁻¹ with the addition of 1.5 g L⁻¹ of sodium acetate and 100 mM 2-(N-morpholino)ethanesulfonic acid (MES), respectively. Especially when 1.5 g L⁻¹ of sodium acetate and 100 mM MES were added simultaneously, the production of hexanoic acid increased up to 2.99 g L⁻¹. Without adding sodium acetate, 2.75 g L⁻¹ of hexanoic acid production from d-galactitol was achieved using a coculture of Clostridium sp. BS-1 and one of the isolates, Clostridium sp. BS-7, in the presence of 100 mM MES. In addition, volatile fatty acid (VFA) production by Clostridium sp. BS-1 from d-galactitol and d-glucose was enhanced when a more reduced culture redox potential (CRP) was applied via addition of Na₂S·9H₂O.     

Influence of initial cellulose concentration on the carbon flow distribution during batch fermentation by Clostridium thermocellum ATCC 27405

The objective of this research was to understand how carbon loading influences hydrogen (H₂) synthesis and metabolic flow patterns in the thermophilic, cellulolytic bacterium, Clostridium thermocellum. C. thermocellum was cultivated in batch cultures with high (5 g L⁻¹) and low (1 g L⁻¹) initial concentrations of α-cellulose at 60°C. The growth rate of C. thermocellum was 22% lower (0.15 h⁻¹) in cultures with low-cellulose concentration compared with cultures with high-cellulose concentrations. Although substrate depletion coincided with the end of log-growth in low-cellulose cultures, the prime reason for growth arrest in high-cellulose cultures was not identified. Ethanol, acetate, and formate were the major soluble end-products with concomitant release of H₂ and CO₂ under both conditions. Lactate appeared during the late log phase in high-carbon cultures when pH dropped below 6.4 and became the major end-product in stationary phase. During the exponential phase of cell growth, significantly higher yields for H₂ and acetate (1.90 ± 0.14 and 1.11 ± 0.04 mol/mol glucose equivalent, respectively) were obtained from low-cellulose cultures compared to those from high-cellulose cultures. The maximum specific rate of H₂ production, 6.41 ± 0.13 mmol H₂/g dry cell/h, obtained during the exponential phase from low-carbon cultures was about 37% higher than that obtained from high-carbon cultures.     

Microbial Succession in a Compost-packed Biofilter Treating Benzene-contaminated Air

Air artificially contaminated with increasing concentrations of benzene was treated in a laboratory scale compost-packed biofilter for 240 days with a removal efficiency of 81–100%. The bacterial community in the packing material (PM) at different heights of the biofilter was analysed every 60 days. Bacterial plate counts and ribosomal intergenic spacer analysis (RISA) of the isolated strains showed that the number of cultivable aerobic heterotrophic bacteria and the species diversity increased with benzene availability. Identification of the isolated species and the main bands in denaturing gradient gel electrophoresis (DGGE) profiles from total compost DNA during the treatment revealed that, at a relatively low volumetric benzene load (1.2≤VBL≤6.4 g m⁻³ _PM h⁻¹), besides low G+C Gram positive bacteria, originally present in the packing compost, bacteroidetes and β- and γ-proteobacteria became detectable in the colonising population. At the VBL value (24.8 g m⁻³ _PM h⁻¹) ensuring the maximum elimination capacity of the biofilter (20.1 g m⁻³ _PM h⁻¹), strains affiliated to the genus Rhodococcus dominated the microflora, followed by β-proteobacteria comprising the genera Bordetella and Neisseria. Under these conditions, more than 35% of the isolated strains were able to grow on benzene as the sole carbon source. Comparison of DGGE and automated RISA profiles of the total community and isolated strains showed that a complex bacterial succession occurred in the reactor in response to the increasing concentrations of the pollutant and that cultivable bacteria played a major role in benzene degradation under the adopted conditions.     

Controlling the oxidoreduction potential of the culture of <Emphasis Type="Italic">Clostridium acetobutylicum</Emphasis> leads to an earlier initiation of solventogenesis,thus increasing solvent productivity

Fermentative production of solvents (acetone, butanol, and ethanol) by Clostridium acetobutylicum is generally a biphasic process consisting of acidogenesis and solventogenesis. We report that the biphasic metabolism of C. acetobutylicum could be changed by oxidoreduction potential (ORP) regulation. When using air to control the ORP of the fermentation broth at −290 mV, an earlier initiation of solventogenesis was achieved. Solvent production reached 25.6 g l⁻¹ (2.8 g acetone l⁻¹, 16.8 g butanol l⁻¹, 6.0 g ethanol l⁻¹), a 35% increase compared with the ORP uncontrolled process. Metabolic flux analysis revealed that there was a general increase of the central carbon flux in the first 24 h of fermentation when ORP was controlled at −290 mV, compared with the control. Specifically, the solvent ratio (acetone:butanol:ethanol) was changed from 25:64:11 to 11:66:23 at ORP level of −290 mV, which might have resulted from the rigidity at acetyl-CoA node and the flexibility at acetoacetyl-CoA and butyryl-CoA nodes in response to ORP regulation.     

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.     

Kinetics of lipase catalyzed isoamyl acetate synthesis at high hydrostatic pressure in hexane

Lipase-catalyzed synthesis of isoamyl acetate in hexane at 10–250 MPa at 80°C and 1–100 MPa at 40°C resulted in activation volumes of −12.9 ± 1.7 and −21.6 ± 2.9 cm³ mol⁻¹, respectively. Increasing pressure from 10 to 200 MPa resulted in approximately 10-fold increase in V _max at both 40 and 80°C. Pressure increased the K _m from 2.4 ± 0.004 to 38 ± 0.78 mM at 40°C. In contrast, at 80°C the pressure did not affect the K _m.     

Biodegradation of methyl <Emphasis Type="Italic">tert</Emphasis>-butyl ether by cold-adapted mixed and pure bacterial cultures

An aerobic mixed bacterial culture (CL-EMC-1) capable of utilizing methyl tert-butyl ether (MTBE) as the sole source of carbon and energy with a growth temperature range of 3 to 30°C and optimum of 18 to 22°C was enriched from activated sludge. Transient accumulation of tert-butanol (TBA) occurred during utilization of MTBE at temperatures from 3°C to 14°C, but TBA did not accumulate above 18°C. The culture utilized MTBE at a concentration of up to 1.5 g l⁻¹ and TBA of up to 7 g l⁻¹. The culture grew on MTBE at a pH range of 5 to 9, with an optimum pH of 6.5 to 7.1. The specific growth rate of the CL-EMC-1 culture on 0.1 g l⁻¹ of MTBE at 22°C and pH 7.1 was 0.012 h⁻¹, and the growth yield was 0.64 g (dry weight) g⁻¹. A new MTBE-utilizing bacterium, Variovorax paradoxus strain CL-8, isolated from the mixed culture utilized MTBE, TBA, 2-hydroxy isobutyrate, lactate, methacrylate, and acetate as sole sources of carbon and energy but not 2-propanol, acetone, methanol, formaldehyde, or formate. Two other isolates, Hyphomicrobium facilis strain CL-2 and Methylobacterium extorquens strain CL-4, isolated from the mixed culture were able to grow on C₁ compounds. The combined consortium could thus utilize all of the carbon of MTBE.     

Induction of multiple shoots in a monopodial orchid hybrid (Aerides vandarum Reichb.f × Vanda stangeana Reichb.f) using thidiazuron and analysis of their genetic stability

The effect of thidiazuron (TDZ) on direct multiple shoot induction in axenic seedlings of a monopodial orchid hybrid Aerides vandarum × Vanda stangeana, using a dual phase culture medium was studied. The culture system consisted of a basal agar solidified half-strength Murashige and Skoog medium overlaid by a liquid fraction of the same composition. Highest regeneration of multiple shoots (15.8 shoots per seedling) was obtained in the medium containing 2% sucrose (w/v) supplemented with 2 mgl⁻¹ TDZ. Genetic stability of the regenerated shoots was assessed using random amplified polymorphic DNA (RAPD), inter simple sequence repeat (ISSR), and restriction fragment length polymorphism of the PCR amplified (PCR-RFLP) nrITS region, as well as those of the coding (matK) and non-coding (trnL-F) regions of the cpDNA. Across the randomly selected mother plant and nine of its regenerated shoots, 2,680 bands were generated by 19 RAPD and 12 ISSR primers, exhibiting monomorphic banding profiles. Homogenous PCR-RFLP profiles were generated for nrITS using four restriction enzymes (REs), matK using five REs, and trnL-F using six REs. These molecular analyses showed no genomic alterations in all regenerated shoots obtained on medium containing 2 mgl⁻¹ TDZ.     

Biological nitrification–denitrification with alternating oxic and anoxic operations using aerobic granules

Efficient nitrification and denitrification of wastewater containing 1,700 mgl⁻¹ of ammonium-nitrogen was achieved using aerobic granular sludge cultivated at medium-to-high organic loading rates. The cultivated granules were tested in a sequencing batch reactor (SBR) fed with 6.4 or 10.2 kg NH₄⁺-N m⁻³ day⁻¹, a loading significantly higher than that reported in literature. With alternating 2 h oxic and 2 h anoxic operation (OA) modes, removal rate was 45.5 mg NH₄⁺-N g⁻¹ volatile suspended solids⁻¹ h⁻¹ at 6.4 kg NH₄⁺-N m⁻³ day⁻¹ loading and 41.3 ± 2.0 at 10.2 kg NH₄⁺-N m⁻³ day⁻¹ loading. Following the 60 days SBR test, granules were intact. The fluorescence in situ hybridization and confocal laser scanning microscopy results indicate that the SBR-OA granules have a distribution with nitrifers outside and heterotrophs outside that can effectively expose functional strains to surrounding substrates at high concentrations with minimal mass transfer limit. This microbial alignment combined with the smooth granule surface achieved nitrification–denitrification of wastewaters containing high-strength ammonium using aerobic granules. Conversely, the SBR continuous aeration mode yielded a distribution with nitrifers outside and heterotrophs inside with an unsatisfactory denitrification rate and floating granules as gas likely accumulated deep in the granules.     

Age,growth, and population structure of the smooth clam <Emphasis Type="Italic">Callista chione</Emphasis> in the eastern Adriatic Sea

The age, growth, and population structure of the smooth clam Callista chione were determined from samples collected by hydraulic dredge and SCUBA at four locations in the eastern Adriatic during 2007 and 2008. The age of 436 clam shells was determined from internal growth lines present in shell sections, and the timing of growth line formation was ascertained from monthly collections of clams to occur between August and September when sea water temperatures were maximal. In addition, age of 30 older individuals was verified with acetate peels of polished and etched shell sections. Differences were apparent in the age structure and growth rates of clams collected from the four locations studied. Von Bertalanffy growth (VBG) curves obtained for clams from these locations were L _t  = 72.4 (1−e^{−0.25(t − 2.68)}) (Rab Island), L _t  = 74.5 (1−e^{−0.15(t + 0.57)}) (Pag Bay), L _t  = 79.3 (1−e^{−0.34(t − 0.97)}) (Cetina estuary), and L _t  = 82.5 (1−e^{−0.11(t + 2.88)}) (Kaštela Bay). The age of the clams ranged between 3 and 44 years; median clam ages were similar at three of the four locations (14, 12, and 12 years, respectively), but was significantly lower in the Cetina estuary (4 years). The VBG growth constants recorded from clams were within the range of values obtained for this species by previous authors. The observed local differences in population structure indicate different levels of exploitation and illustrate the need to establish long-term strategies for a sustainable exploitation of smooth clams in the Croatian Adriatic.     

Biochemical kinetic behaviors between n-butyl acetate and composite bead in biofilter

In this study, the kinetic behaviors between n-butyl acetate and composite bead were investigated. Both microbial growth rate and biochemical reaction rate would be inhibited with increasing average inlet concentration. The order of the inhibitive effect, which resulted from increased average inlet concentration for four operation temperatures, was 30>35>40>25 °C. Both microbial growth rate and biochemical reaction rate would be enhanced and inhibited with increasing operation temperature in the operation temperature ranges of 25 to 30 and 30 to 40 °C, respectively. The enhancing and inhibitive effects resulting from increased operation temperature were the most pronounced at the average inlet concentration of 200 ppm. The values of maximum reaction rate V _m and half-saturation constant K _s ranged from 0.011 to 0.047 g C h⁻¹ kg⁻¹ packed material and from 19.30 to 62.40 ppm, respectively. The zero-order kinetic with the diffusion rate limitation could be regarded as the most adequate biochemical reaction kinetic model. The values of maximum elimination capacity ranged from 0.51 to 0.20 g C h⁻¹ kg⁻¹ packed material, and the optimal maximum elimination capacity of biofilter occurred at the operation temperature of 30 °C.     

Effects of manganese on the growth,photosystem II and SOD activity of the dinoflagellate <Emphasis Type="Italic">Amphidinium</Emphasis> sp.

Experimental ecology methods and chlorophyll fluorescence technology were used to study the effects of different concentrations of manganese (10⁻¹²– 10⁻⁴ mol L⁻¹) on the growth, photosystem II and superoxide dismutase (SOD) activity of Amphidinium sp. MACC/D31. The results showed that manganese had a significant effect on the growth rate, fluorescence parameters (maximal photochemical efficiency of PSII (F _v /F _m ), photochemical quenching (qP) and non-photochemical quenching (NPQ)) in the exponential stage (days 1–3) and SOD activity of Amphidinium sp. (P < 0.05). F _v/F _m in the exponential stage in 10⁻¹² mol L⁻¹ manganese concentration was significantly lower whilst qP and NPQ significantly higher than those in the other concentrations. F _v /F _m (days 6–9) in 10⁻⁴ mol L⁻¹ manganese was significantly higher than those in the other concentrations. F _v /F _m (days 3–6) increased with increased concentration of manganese from 10⁻¹² to 10⁻⁴ mol L⁻¹. The values of qP and NPQ decreased with decreased concentrations of manganese, except for those in days 4–6. F _v /F _m under each concentration increased earlier and decreased later with culture stage whilst NPQ decreased earlier and increased later. The SOD activity increased with increased concentration of manganese from 10⁻¹² to 10⁻⁸ mol L⁻¹. The SOD activity in 10⁻⁴ mol L⁻¹ manganese was significantly higher than those in the other concentrations and in 10⁻¹² mol L⁻¹ manganese, it was significantly lower than those in the other concentrations.     

Effects of nitrogen source and empty bed residence time on the removal of styrene gaseous emissions by biotrickling filtration

The removal of styrene-polluted air emissions by biotrickling filtration was performed to evaluate the influence of using nitrate and urea as a nitrogen source in the nutrient solution supplied to two bioreactors run in parallel under the same operational conditions for 3 months. The use of urea resulted in less biomass content along the packed bed and better performance of the process, with a maximum elimination capacity (EC) of 57.6 g C m⁻³ h⁻¹ (removal efficiency (RE) of 88.3% and empty bed residence time (EBRT) of 60 s), which was around 54% higher than when using nitrate. EBRTs of 60, 30 and 15 s were evaluated with a urea-based nutrient supply. By decreasing the EBRT from 60 to 30 s the styrene concentration that could be treated with REs above 80% was almost the half, from 1,100 to 600 mg C m⁻³, resulting in ECs of 52.8 g C m⁻³ h⁻¹. Working at 15 s was not possible to obtain REs higher than 40% with a maximum EC of 28.5 g C m⁻³ h⁻¹.     

<Emphasis Type="Italic">Clostridium beijerinckii</Emphasis> mutant with high inhibitor tolerance obtained by low-energy ion implantation

Clostridium beijerinckii mutant strain IB4, which has a high level of inhibitor tolerance, was screened by low-energy ion implantation and used for butanol fermentation from a non-detoxified hemicellulosic hydrolysate of corn fiber treated with dilute sulfuric acid (SAHHC). Evaluation of toxicity showed C. beijerinckii IB4 had a higher level of tolerance than parent strain C. beijerinckii NCIMB 8052 for five out of six phenolic compounds tested (the exception was vanillin). Using glucose as carbon source, C. beijerinckii IB4 produced 9.1 g l⁻¹ of butanol with an acetone/butanol/ethanol (ABE) yield of 0.41 g g⁻¹. When non-detoxified SAHHC was used as carbon source, C. beijerinckii NCIMB 8052 grew well but ABE production was inhibited. By contrast, C. beijerinckii IB4 produced 9.5 g l⁻¹ of ABE with a yield of 0.34 g g⁻¹, including 2.2 g l⁻¹ acetone, 6.8 g l⁻¹ butanol, and 0.5 g l⁻¹ ethanol. The remarkable fermentation and inhibitor tolerance of C. beijerinckii IB4 appears promising for ABE production from lignocellulosic materials.