Biodegradation of phenanthrene and pyrene by Ganoderma lucidum

Biodegradation of two polycyclic aromatic hydrocarbons (PAHs), phenanthrene and pyrene, by a white rot fungus, Ganoderma lucidum, in broth cultures was investigated. It was found that the biomass of the organism decreased with the increase of PAH concentration in the cultures. In the cultures with 2 to 50 mg l⁻¹ PAHs, the degradation rate constants (k₁) increased with the PAH concentration, whereas, at the level of 100 mg l⁻¹, the degradation rate constants decreased. In the presence of 20 mg l⁻¹ PAHs, the highest degradation rates of both PAHs occurred in cultures with an initial pH of 4.0 at 30 °C. The addition of CuSO₄, citric acid, gallic acid, tartaric acid, veratryl alcohol, guaiacol, 2,2′-azino-bis-(3- ethylbenzothazoline-6-sulfonate) (ABTS) enhanced the degradation of both PAHs and laccase activities; whereas the supplement of oxalate, di-n-butyl phthalate (DBP), and nonylphenol (NP) decreased the degradation of both PAHs and inhibited laccase production. In conclusion, G. lucidum is a promising white rot fungus to degrade PAHs such as phenanthrene and pyrene in the environment.     

Pentachlorophenol (PCP) dechlorination in horizontal-flow anaerobic immobilized biomass (HAIB) reactors

This study verifies the potential applicability of horizontal-flow anaerobic immobilized biomass (HAIB) reactors to pentachlorophenol (PCP) dechlorination. Two bench-scale HAIB reactors (R1 and R2) were filled with cubic polyurethane foam matrices containing immobilized anaerobic sludge. The reactors were then continuously fed with synthetic wastewater consisting of PCP, glucose, acetic acid, and formic acid as co-substrates for PCP anaerobic degradation. Before being immobilized in polyurethane foam matrices, the biomass was exposed to wastewater containing PCP in reactors fed at a semi-continuous rate of 2.0 μg PCP g⁻¹ VS. The applied PCP loading rate was increased from 0.05 to 2.59 mg PCP l⁻¹ day⁻¹ for R1, and from 0.06 to 4.15 mg PCP l⁻¹ day⁻¹ for R2. The organic loading rates (OLR) were 1.1 and 1.7 kg COD m⁻³ day⁻¹ at hydraulic retention times (HRT) of 24 h for R1 and 18 h for R2. Under such conditions, chemical oxygen demand (COD) removal efficiencies of up to 98% were achieved in the HAIB reactors. Both reactors exhibited the ability to remove 97% of the loaded PCP. Dichlorophenol (DCP) was the primary chlorophenol detected in the effluent. The adsorption of PCP and metabolites formed during PCP degradation in the packed bed was negligible for PCP removal efficiency.     

Versatility of fluorene metabolite (phenol) in fluorene biodegradation by a sulfate reducing culture

Biodegradability of fluorene and the versatility of fluorene metabolite (i.e. phenol) in fluorene biodegradation by a sulfate-reducing enrichment culture were investigated. Batch experiments (with 5 mg l⁻¹ fluorene) were designed via the central composite design to examine the effects of sulfate (5-35 mM) and biomass (5-50 mg l⁻¹) concentrations (variables) on fluorene degradation (response). The experimental results revealed that fluorene removal was more influenced by the biomass concentration than the sulfate concentration. The optimal sulfate and biomass concentrations for fluorene biodegradation (90% removal) were found to be 14.4 mM and 37.8 mg l⁻¹, respectively. Under the optimal conditions, a set of biodegradation experiments were repeated to evaluate both the biodegradability of fluorene metabolite and the potential effect of phenol accumulation on fluorene degradation. The outcomes indicated a slow phenol degradation rate, i.e. 0.02 mg l⁻¹ d⁻¹. Moreover, a small reduction in the fluorene biodegradation efficiency was observed in the presence and accumulation of phenol. However, this sulfate reducing culture is a valuable resource for the simultaneous degradation of fluorene and phenol.     

Denitrifying sulfide removal and carbon methanogenesis in a mesophilic, methanogenic culture

The inhibitory effects of 90-189 mg l⁻¹ of sulfide and 25-75 mg-N l⁻¹ of nitrate on methanogenesis were investigated in a mixed methanogenic culture using butyrate as carbon source. In the initial phase of 90 mg l⁻¹ S²⁻ test, autotrophic denitrification of nitrate occurred with sulfide as the electron donor. Then the sulfate-reducing strains converted the produced sulfur back to sulfide via heterotrophic oxidation pathway. Methanogenesis was not markedly inhibited when 90 mg l⁻¹ of sulfide was dosed alone. When 25-75 mg-N l⁻¹ of nitrate was presented, initiation of methanogenesis was seriously delayed. Nitrogen oxides (NO_x), the intermediates for nitrate reduction via denitrification pathway, inhibited methanogenesis. The 90 mg l⁻¹ of sulfide favored heterotrophic dissimilatory nitrate reduction to ammonia (DNRA) pathway for nitrate reduction. Possible ways of maximizing methane production from an organic carbon-rich wastewater with high levels of sulfide and nitrate were discussed.     

Sulfide enhances methanogenesis in nitrate-containing methanogenic cultures

The effects of sulfide on nitrate reduction and methanogenesis using butyrate as a carbon source were investigated in a mixed mesophilic, methanogenic culture. In the sulfide-free medium, 25-75 mg l⁻¹ nitrate markedly inhibited the efficiencies of acetogenesis and methanogenesis processes. Adding 25 mg-S l⁻¹ increased methane production in nitrate-amended medium. Low sulfide levels shifted the nitrate reduction pathway from denitrification to dissimilatory nitrate reduction to ammonia (DNRA), thereby reducing the amounts of toxic nitric oxide and nitrous oxide produced that inhibit methanogenesis. The dose of 25 mg l⁻¹ sulfide was oxidized completely, during which heterotrophic DNRA predominated. The oxidized forms of sulfide reformed, limiting induction of the heterotrophic denitrification pathway. The actions of heterotrophic and autotrophic DNRA bacteria, denitrifiers, sulfate-reducing bacteria and methanogens mitigate nitrate toxicity during methanogenesis in an anaerobic process.     

Biodegradation of phenol using Corynebacterium sp. DJ1 aerobic granules

The single-culture Corynebacterium sp. DJ1 aerobic granules were cultivated and were utilized to degrade high-strength phenolic wastewater. These granules can degrade phenol at sufficient high rate without severe inhibitory effects up to phenol concentration of 2000 mg l⁻¹. Furthermore, the kinetic characteristic noted for these granules yields a zero-order phenol degradation behavior with 500–1500 mg l⁻¹ phenol, which facilitates reactor design and scale up. With added acetate to promote cell growth, this single-culture aerobic granular system yields the highest phenol degradation rate reported in granular literature.     

An in vitro propagation protocol of two submerged macrophytes for lake revegetation in east China

Thirty-six programs have been set up to revegetate the degraded lake wetlands in east China since 2002. Most projects however faced deficiency of submerged macrophyte propagules. To solve the problem, alternative seedling sources must be found besides traditional field collection. This paper deals with an in vitro propagation protocol for two popularly used submerged macrophytes, Myriophyllum spicatum L. and Potamogeton crispus L. Full strength Murashige and Skoog-based liquid media (MS) plus 3% sucrose in addition to 0–2.0 mg l⁻¹ 6-benzylaminopurine (BA) and 0–1.0 mg l⁻¹ indoleacetic acid (IAA) were tried for shoot regeneration. Meanwhile, full, half or quarter strength MS in addition to 0, 0.1 or 0.2 mg l⁻¹ naphthaleneacetic acid (NAA) were tested for root induction, respectively. Results indicated that both species had the ability of regeneration from stem fragments in MS without further regulators. However, the addition of 2.0 mg l⁻¹ BA with 0.2 or 1.0 mg l⁻¹ IAA in MS drastically stimulated the regeneration efficiency of M. spicatum, while the addition of 2.0 mg l⁻¹ BA with 0.2 or 0.5 mg l⁻¹ IAA in MS significantly stimulated that of P. crispus. For root induction, full strength MS in combination with 0.1or 0.2 mg l⁻¹ NAA was preferred by M. spicatum, and the same MS without or with 0.1 mg l⁻¹ NAA was preferred by P. crispus. Seedlings of each species produced from tissue culture room had a 100% survival rate on clay, sandy loam or their mixture (1:1) in an artificial pond, and phenotypic plasticity was exhibited when the nutrient levels varied among the three types of sediments. This acclimation of seedlings helped develop the shoot and root systems, which ensured seedling quality and facilitated the transplantation. Our study has established an effective protocol to produce high quality seedlings for lake revegetation programs at a larger scale. Since the two species we tested represent different regeneration performances in nature but shared similar in vitro propagation conditions, this study has indicated a potentially wide use of the common media for preparing seedlings of other submerged macrophytes.     

Characterization of alkaline thermostable keratinolytic protease from thermoalkalophilic Bacillus halodurans JB 99 exhibiting dehairing activity

A thermostable alkaline protease produced from Bacillus sp. JB 99 exhibited significant keratinolytic and dehairing activity. The enzyme was purified by ammonium sulphate precipitation followed by CM-cellulose and Sephadex G-100 chromatography and resulted in 13.6 fold purification with 23.8% of recovery. The specific activity of purified enzyme was 2989.6 U mg^−l. Purified protease had a molecular weight of 29 kDa and appeared as a single band. Gelatin zymogram analysis also revealed a clear hydrolytic zone, which corresponded to the band obtained with SDS-PAGE. The optimum pH and temperature for the keratinolytic activity was pH 11.0 and 70 °C respectively and half life of protease was 70 °C for 4 h. N-terminal amino acid sequence of purified enzyme exhibited extensive homology with other thermostable alkaline proteases and inhibition by PMSF indicated serine type of protease. The K_m and V_max of protease for keratin substrate were 3.8 ± 0.5 mg ml⁻¹ and 15.1 ± 1.6 ??m min⁻¹ mg⁻¹ and casein were 3.3 ± 0.4 mg ml^−l and 15.6 ± 0.9 ??m min⁻¹ mg⁻¹ respectively. The enzyme efficiently dehaired buffalo and goat hide without damaging the collagen layer, which makes it a potential candidate for application in leather industry to avoid pollution problem associated with the use of chemicals in the industry. The enzyme also degraded chicken feathers in presence of reducing agent which can help poultry industry in management of keratin-rich waste and obtaining value added products.     

Laboratory scale examination of the effects of overloading on the anaerobic digestion by glycerol

The anaerobic digestion of pure glycerol, which produces a baseline acetic acid to propionic acid ratio of 0.2, was studied in laboratory scale reactors (3 l working volume) at mesophilic temperature (37 °C) with 3000 mg chemical oxygen demand (COD) l⁻¹d⁻¹. During the experiment tVFA and C2-C6 VFA analysis and daily biogas yield measurement were carried out. Following 10 days of a 15% d⁻¹ increase in the organic loading rate (OLR) of 3.0-10.5 g COD l⁻¹d⁻¹, the concentration of propionic acid increased to 6200-8000 mg l⁻¹. Then the inoculum was divided into three parts feeding with 100% glycerol, 50% glycerol + 50% acetic acid, and 50% glycerol + 50% thick stillage, (presented in % of 2.60 g COD l⁻¹d⁻¹ OLR), respectively. The application of co-substrates reduced the recovery period by 5 days compared to feeding with pure glycerol. When the reactors were loaded with glycerol again (10% OLR raise per day) the previously applied co-substrates had a positive effect on the VFA composition and the biogas yield as well.     

Lovastatin biosynthesis by Aspergillus terreus with the simultaneous use of lactose and glycerol in a discontinuous fed-batch culture

The influence of various combinations of glycerol and lactose feed on the biosynthesis of two polyketide metabolites, lovastatin and (+)-geodin, by Aspergillus terreus ATCC20542 in a discontinuous fed-batch culture was presented. In these experiments lactose and/or glycerol were also used as the initial carbon substrates in the cultivation media. The application of glycerol feed, when lactose is the initial substrate, leads to the appreciable lovastatin concentration in the broth (122.4 mg l⁻¹), nevertheless the abundant (+)-geodin level is at the same time obtained (255.5 mg l⁻¹). The cultures with glycerol as the initial substrate and fed with lactose produce less lovastatin and (+)-geodin. The application of the various combined glycerol and/or lactose feeds allows for improving lovastatin production up to 161.8 mg l⁻¹ and decreases (+)-geodin concentration to 98.7 mg l⁻¹. The analysis of product formation rates and yield coefficients indicates that lovastatin is more efficiently produced on lactose, especially in the initial stages of the cultivation. Glycerol efficiently sustains fungal activity to form these polyketides in the late idiophase but it mainly favours (+)-geodin formation, if solely used in the feed. The feeds performed both with lactose and glycerol occur to be the most desired to maximise lovastatin and minimise (+)-geodin formation.     

A comparison of irradiance and phosphorus effects on the growth of three submerged macrophytes

A fully factorial pond experiment was designed using two irradiance levels and two phosphorus concentrations to investigate irradiance and phosphorus effects on the growth of three submerged macrophytes: common waterweed (Elodea canadensis), Eurasian water milfoil (Myriophyllum spicatum), and water stargrass (Zosterella dubia). Results revealed that higher irradiance (230 μmol s⁻¹ m⁻² vs. 113 μmol s⁻¹ m⁻² at 2 m depth) had significant positive effects on submerged macrophyte growth: increasing the number of individuals (seven-fold), the number of species surviving (two-fold), aboveground biomass (11-fold), belowground biomass (10-fold), and total biomass (11-fold), whereas elevated sediment phosphorus (2.1–3.3 mg g⁻¹ vs. 0.7 mg g⁻¹ dry sediment) did not have any significant impact. However, responses to irradiance differ among macrophyte species due to their morphology and physiology. Waterweed increased in numbers of individuals and total biomass under high irradiance while biomass per individual remained the same (∼0.02 g). The other species increased both in numbers and biomass per individual. These results suggest that increased irradiance rather than decreased phosphorus loading is the main driver of changes in submerged macrophytes in North American temperate lake ecosystems.     

Effects of light regime and oxygen profile on transformation of oxolinic acid in pond sediment

This study investigated the effects of light (visible light - 5800 lux, 24 h) or dark regime and aerobic or anaerobic condition on the decay of added oxolinic acid (OA) at 5, 10 and 20 mg L⁻¹ in eel pond sediment. An asymptotic decaying exponential model C_t = C_min + C_o × exp (−k × t) was used to facilitate quantitative approach to OA transformation, where C_t is the concentration of OA after t days, C_min the estimated level-off concentration of OA residue, C_o the concentration of added OA and k the decaying coefficient. OA decayed faster under light (C_min = 4.6 mg L⁻¹) than under dark (C_min = 7.8 mg L⁻¹) and also decayed faster under aerobic (C_min = 4.0 mg L⁻¹) than under anaerobic condition (C_min = 8.5 mg L⁻¹). C_min increased with C_o. Sundrying and tilling eel pond bottom should be able to reduce OA residue significantly.     

Phosphorus release from sediments in a treatment wetland: Contrast between DET and EPC0 methodologies

Wetlands are capable of reducing nutrient loadings to receiving water bodies, and hence many artificial wetlands have been constructed for wastewater nutrient removal. In this study, diffusive equilibrium in thin films (DETs) and equilibrium phosphorus concentration (EPC₀) analysis were used to examine the role of sediment as a nutrient source or sink in a constructed treatment wetland in summer. The effect of dredging on sediment-water nutrient exchange was also studied. Soluble reactive phosphorus (SRP), ammonium (NH₄⁺) and sulphate (SO₄²⁻) concentration profiles were measured by DET across the sediment-water interface (SWI) in both a settling pond and iris reed bed within the wetland. The SRP concentrations in the sediment pore-waters of the settling pond were extremely high (up to 29,500 μg l⁻¹) near the SWI. This is over an order of magnitude higher than the levels found in the water column, which in turn are over an order of magnitude higher than environmental levels proposed to limit eutrophication in rivers. The profiles demonstrated an average net release of SRP and NH₄⁺ from the settling pond sediment to the overlying water of 58 mg m⁻² d⁻¹ (±32 mg m⁻² d⁻¹ (1 sd)) and 16 mg m⁻² d⁻¹ (±25 mg m⁻² d⁻¹ (1 sd)), respectively. The DET SO₄²⁻ concentration profiles revealed that the sediment was anoxic within 2 cm of the SWI. Dredging of the reed bed made no significant difference to the P release characteristics across the SWI. The EPC₀s were much lower than the SRP concentration of the overlying water, indicating that the sediment had the potential to act as a phosphate sink. The apparent contradiction of the DET and EPC₀ results is attributed to the fact that DET measurements are made in situ, where as EPC₀ measurements are ex situ. These results show that substantial releases of P can occur from wetland sediments, and also highlight the need for caution when interpreting ex situ EPC₀ analytical results.     

Influence of environmental factors on Vallisneria americana seed germination

Over the course of a growing season (April–October) water quality (water temperature, light, salinity, dissolved oxygen) and reproductive phenology (biomass, production of flowering shoots and seed pods, seed bank densities) were quantified in three Vallisneria americana beds in Nanjemoy Creek, MD, a tributary to the Chesapeake Bay. Clonal production of V. americana biomass increased at all sites when water temperatures rose above 25 °C. Flowering occurred during peak biomass (August–September) and resulted in the production of up to 16,000 seeds m⁻² at the end of the growing season. However, observed seed bank densities represented <1% of seed production. Laboratory experiments quantified the effects of dissolved oxygen (0.29–8.00 mg l⁻¹), light (0–160 μmol m² s⁻¹), temperature (13–29 °C), salinity (0.1–17.4 psu), sediment composition (3–86% sand; 0.9–8.3% sediment organic content), and burial depth (0.2–10 cm) on V. americana seed germination. Germination of V. americana seeds was enhanced (greater overall germination and shorter time to germination) under oxygenated conditions (8.00 mg l⁻¹), temperatures >22 °C, salinities of <1 psu, and in sediments composed of ≤3% organic content and >40% sand. Light (<160 μmol m⁻² s⁻¹) and burial depth (0.2–10 cm) had no significant effects on germination. Temperatures most favorable for seed germination (>22 °C) occurred in June, 2 months in the growing season just prior to development of peak vegetative standing stock. Seedlings were therefore at a distinct disadvantage to plants developed from over wintering buds. A lack of viable seed retention and inadequate environmental conditions at critical times in the growing season may be limiting seed germination success and subsequent seedling establishment within V. americana beds in the Chesapeake Bay. However, ungerminated seeds were found to maintain high viability, especially at salinities of 10 psu that can have significant negative effects of shoot growth survival. This suggests that seeds may serve as a source of reproductive material for bed recovery after periods of drought or other stressful conditions in estuarine systems.     

Performance of a pilot-scale sewage treatment: an up-flow anaerobic sludge blanket (UASB) and a down-flow hanging sponge (DHS) reactors combined system by sulfur-redox reaction process under low-temperature conditions

Performance of a wastewater treatment system utilizing a sulfur-redox reaction of microbes was investigated using a pilot-scale reactor that was fed with actual sewage. The system consisted of an up-flow anaerobic sludge blanket (UASB) reactor and a down-flow hanging sponge (DHS) reactor with a recirculation line. Consequently, the total COD_Cr (465 ± 147 mg L⁻¹; total BOD of 207 ± 68 mg L⁻¹) at the influent was reduced (70 ± 14 mg L⁻¹; total BOD of 9 ± 2 mg L⁻¹) at the DHS effluent under the conditions of an overall hydraulic retention time of 12 h, a recirculation ratio of 2, and a low-sewage temperature of 7.0 ± 2.8 °C. A microbial analysis revealed that sulfate-reducing bacteria contributed to the degradation of organic matter in the UASB reactor even in low temperatures. The utilized sulfur-redox reaction is applicable for low-strength wastewater treatment under low-temperature conditions.     

Molecular cloning,characterization and mRNA expression of selenium-dependent glutathione peroxidase from abalone Haliotis discus hannai Ino in response to dietary selenium,zinc and iron

A novel selenium-dependent glutathione peroxidase (Se-GPX) was cloned from abalone Haliotis discus hannai Ino (HdhGPx) by homology cloning with degenerate primers and RACE techniques. The full length of HdhGPx cDNA was 963 bp with a 669 bp open reading frame (ORF) encoding 222 amino acids and a 101 bp eukaryotic selenocysteine insertion sequence (SECIS) in 3′ untranslated region (UTR). It was showed that HdhGPx has a characteristic codon at ²³⁵TGA²³⁷ that corresponds to selenocysteine (SeC) as U₇₂. Sequence characterization revealed that HdhGPx contains a characteristic GPx signature motif 2 (₉₆LGLPCNQF₁₀₃), an active site motif (₁₇₉WNFEKF₁₈₄). In addition, two potential N-glycosylation sites (₁₁₂NGTE₁₁₅ and ₁₃₂NLTQ₁₃₅) were identified in HdhGPx. 3D modeling analysis showed that the overall structure of HdhGPx monomer had more similarity to human GPx3 than human GPx1. Relatively higher-level mRNA expression was detected in hepatopancreas, mantle and gonad by real-time PCR assays. The relative expression levels of HdhGPx mRNA in hepatopancreas and haemocytes were detected by real-time PCR in abalone fed with nine different diets containing graded levels of selenium (0.15, 1.32 and 48.7 mg kg^− 1), zinc (6.69, 33.85 and 710.63 mg kg^− 1) and iron (29.17, 65.7 and 1267.2 mg kg^− 1) for 20 weeks, respectively. The results showed that the expressions of HdhGPx mRNA were statistically higher at adequate dietary selenium (1.32 mg kg^− 1), zinc (33.85 mg kg^− 1) and iron (65.7 mg kg^− 1) than those in low dietary minerals, respectively. But HdhGPx mRNA expression levels were down-regulated by high contents of dietary selenium (48.7 mg kg^− 1), zinc (710.63 mg kg^− 1) and iron (1267.2 mg kg^− 1), respectively. These results indicated that adequate dietary minerals could increase the mRNA expression of HdhGPx, and then to increase the total antioxidant capacities in abalone.     

Ecology of aquatic Ranunculus communities under the Mediterranean climate

The Iberian Peninsula encompasses more than 80% of the species richness of European aquatic ranunculi. The floristic diversity of the phytocoenosis characterised by aquatic Ranunculus and the main physical–chemical factors of the water were studied in 43 localities of the central Iberian Peninsula. Four aquatic Ranunculus communities are found in most of the aquatic environments. These are species-poor and have an uneven distribution: three species of Batrachium are heterophyllous and their communities are distributed in different aquatic ecosystems on silicated substrates; one species is homophyllous and its community occurs in various aquatic ecosystems with carbonated waters. In the Mediterranean climate, Ranunculus species are present in different habitats, as shown by the results of all the statistical analyses. Ranunculus trichophyllus communities occur in base-rich waters with a high buffering capacity (2273.44 ± 794.57 mg CaCO₃ L⁻¹) and a high concentration of cations (Ca²⁺, 121 ± 33.12 mg L⁻¹; Mg²⁺, 71.64 ± 82.77 mg L⁻¹), nitrates (2.89 ± 4.80 mg L⁻¹), ammonium (2.19 ± 1.36 mg L⁻¹) and sulphates (216.25 ± 218.54 mg L⁻¹). Ranunculus penicillatus communities are found in flowing waters with a high concentration of phosphates (0.48 ± 0.6 mg L⁻¹) and intermediate buffering capacity (683.66 ± 446.76 mg CaCO₃ L⁻¹). Both Ranunculus pseudofluitans and Ranunculus peltatus communities grow in waters with low buffering capacity (R. pseudofluitans, 385.91 ± 209.2 mg CaCO₃ L⁻¹; R. peltatus, 263.3 ± 180.36 mg CaCO₃ L⁻¹), and a low concentration of cations (R. pseudofluitans: Ca²⁺, 12.57 ± 9.42 mg L⁻¹; Mg²⁺, 3.42 ± 1.67 mg L⁻¹; R. peltatus: Ca²⁺, 15 ± 18.26 mg L⁻¹; Mg²⁺, 6.26 ± 8.89 mg L⁻¹) and nutrients (R. pseudofluitans: nitrates, 0.23 ± 0.2 mg L⁻¹; phosphates, 0.09 ± 0.1 mg L⁻¹; R. peltatus: nitrates, 0.19 ± 0.21 mg L⁻¹; phosphates, 0.09 ± 0.12 mg L⁻¹); the first in flowing waters, the latter in still waters.     

Optimising the biogas production from leather fleshing waste by co-digestion with MSW

Waste from the leather industry, known as limed leather fleshing (LF), has a low C:N (3.2) and an alkaline pH of 11.4. This is a major disadvantage for anaerobic digestion due to ammonia toxicity for methanogenesis. This study describes co-digestion of LF with biodegradable fraction of municipal solids waste optimised over a range of C:N and pH to minimise ammonia and to maximise biogas yield. The optimum conditions were found with a blend that provided C:N of 15 and pH of 6.5 and the cumulative biogas yield increased from 560 mL using LF fraction alone, to 6518 mL with optimum blend. At higher pH of 8.5, unionised ammonia was high (2473 mg L⁻¹) coincided with poor biogas yield (47 mL d⁻¹) that confirms ammonia toxicity. By contrast at a pH of 4.5 the ammonia was minimum (510 mg L⁻¹), but high VFA (26,803 mg L⁻¹) inhibited the methanogens. Biomass activity measured using ATP correlated well with biogas yield as reported previously.     

Improved volatile fatty acids production from proteins of sewage sludge with anthraquinone-2,6-disulfonate (AQDS) under anaerobic condition

Organic matters in sewage sludge can be converted into volatile fatty acids (VFAs) as renewable carbon sources. This work for the first time applied anthraquinone-2,6-disulfonate (AQDS) for enhancing VFA production from sewage sludge. With 0.066 or 0.33 g AQDS g⁻¹ dried solids (DS), the yields for VFAs peak at 403 or 563 mg l⁻¹, 1.9- or 2.7-fold to the control. The accumulated VFAs were principally composed of acetate and propionate. The AQDS enhances degradation rates of model proteins (bovine serum albumin), but had little enhancement on that of model polysaccharides (dextrans). The acidification step is proposed the rate-limiting step for VFA production from sewage sludge, in which the AQDS molecules shuttle electrons to accelerate the redox reactions associated with amino acid degradation. Methanogenic activities are inhibited in the presence of AQDS. The AQDS-assisted VFAs are renewable organic carbon sources, although their direct use for anaerobic digestion is not advised.     

Comparison of polysialic acid production in Escherichia coli K1 during batch cultivation and fed-batch cultivation applying two different control strategies

The polysialic acid (PSA) production in Escherichia coli (E. coli) K1 was studied using three different cultivation strategies. A batch cultivation, a fed-batch cultivation at a constant specific growth rate of 0.25 h⁻¹ and a fed-batch cultivation at a constant glucose concentration of 50 mg l⁻¹ was performed. PSA formation kinetics under different cultivation strategies were analyzed based on the Monod growth model and the Luedeking-Piret equation. The results revealed that PSA formation in E. coli K1 was completely growth associated, the highest specific PSA formation rate (0.0489 g g⁻¹ h⁻¹) was obtained in the batch cultivation. However, comparing biomass and PSA yields on the glucose consumed, both fed-batch cultivations provided higher yields than that of the batch cultivation and acetate formation was prevented. Moreover, PSA yield on glucose was also correlated to the specific growth rate of the cells. The optimal specific growth rate for PSA production was 0.32 h⁻¹ obtained in the fed-batch cultivation at a constant glucose concentration of 50 mg l⁻¹, with highest conversion efficiency of 43 mg g⁻¹.