Enhancing Roxarsone Degradation and In Situ Arsenic Immobilization Using a Sulfate-Mediated Bioelectrochemical System.

Roxarsone (ROX) is widely used in animal farms, thereby producing organoarsenic-bearing manure/wastewater. ROX cannot be completely degraded and nor can its arsenical metabolites be effectively immobilized during anaerobic digestion, potentially causing arsenic contamination upon discharge to the environment. Herein, we designed and tested a sulfate-mediated bioelectrochemical system (BES) to enhance ROX degradation and in situ immobilization of the released inorganic arsenic. Using our BES (0.5 V voltage and 350 µM sulfate), ROX and its metabolite, 4-hydroxy-3-amino-phenylarsonic acid (HAPA), were completely degraded within 13-22 days. In contrast, the degradation efficiency of ROX and HAPA was <85% during 32-day anaerobic digestion. In a sulfate-mediated BES, 75.0-83.2% of the total arsenic was immobilized in the sludge, significantly more compared to the anaerobic digestion (34.1-57.3%). Our results demonstrate that the combination of sulfate amendment and voltage application exerted a synergetic effect on enhancing HAPA degradation and sulfide-driven arsenic precipitation. This finding was further verified using real swine wastewater. A double-cell BES experiment indicated that As(V) and sulfate were transported from the anode to the cathode chamber and coprecipitated as crystalline alacranite in the cathode chamber. These findings suggest that the sulfate-mediated BES is a promising technique for enhanced arsenic decontamination of organoarsenic-bearing manure/wastewater.

Electrochemical stimulation of microbial roxarsone degradation under anaerobic conditions.

Roxarsone (4-hydroxy-3-nitrophenylarsonic acid) has been commonly used in animal feed as an organoarsenic additive, most of which is excreted in manure. Roxarsone is easily biodegraded to 4-hydroxy-3-aminophenylarsonic acid (HAPA) under anaerobic conditions, but HAPA persists for long periods in the environment, increasing the risk of arsenic contamination through diffusion. We investigated the electrochemical stimulation of the microbial degradation of roxarsone under anaerobic conditions. After the carbon sources in the substrate were depleted, HAPA was slowly degraded to form arsenite under anaerobic conditions. The degradation rate of HAPA was significantly increased when 0.5 V was applied without adding a carbon source. The two-cell membrane reactor assays reveal that the HAPA was degraded in the anode chambers, confirming that the anode enhanced the electron transfer process by acting as an electron acceptor. The degradation product formed with electrochemical stimulation was arsenate, which facilitates the removal of arsenic from wastewater. Based on the high performance liquid chromatography-ultraviolet-hydride generation-atomic fluorescence spectrometry (HPLC-UV-HG-AFS) and gas chromatography-mass spectrometry (GC-MS) data, the pathway for the biodegradation of roxarsone and the mechanisms for the electrochemically stimulated degradation are proposed. This method provides a potential solution for the removal of arsenic from organoarsenic-contaminated wastewater.

Ca(ClO)2 pretreatment enhancing suspended solids removal through flocculation from digested dairy wastewater and its mechanisms.

Intensive animal farming produces large volume of digested liquid, and overdose application often causes the pollution of surface water and groundwater. Therefore, post-treatment is very necessary for the discharging of surplus digested liquid, but the removal of high concentrations of suspended solids (SS) in the digested liquid is a challenge. In this study, the effect of Ca(ClO)2 pretreatment on SS flocculation removal of digested dairy wastewater was investigated. The results showed that, without Ca(ClO)2 pretreatment, the flocculation by polyacrylamide (PAM), polyferric sulfate (PFS) or polymeric aluminum chloride (PAC) only removed 42.6 %-50.4 % SS from anaerobic digested liquid. With the combination of Ca(ClO)2 pretreatment and PAC flocculation together, the SS removal efficiency can reach 80 %. The total chemical oxygen demand (TCOD) removal had a similar trend with SS removal, but soluble chemical oxygen demand (SCOD) removal was less affected by the pretreatment and flocculation. More than 75 % of orthophosphate (SRP) and total soluble phosphorus (TSP) was removed after Ca(ClO)2 pretreatment and flocculation with PFS or PAC. Ca(ClO)2 pretreatment also effectively inactivated fecal bacteria. The mechanisms of Ca(ClO)2 pretreatment enhancing SS flocculation removal were further elucidated. The SS removal was the action of ClO- and Ca2+ together. The function of ClO- was to break down suspended particles, change the surface, and decrease the absolute Zeta potential, while the function of Ca2+ was to form precipitation. This result indicates that Ca(ClO)2 pretreatment can effectively enhance the SS flocculation removal of anaerobic digested liquid.

Modification of fluorescence staining method for small-sized microplastic quantification: Focus on the interference exclusion and exposure time optimization.

Microplastics are an emerging pollutant of global concern, and fluorescence staining as an efficient method for small-sized microplastic qualification often undergoes the serious interference from external environments. The key steps affecting the accuracy of fluorescent staining and the corresponding quality assurance measures were rarely known. Therefore, this study took the Nile Red/DAPI co-staining method as an example to explore the key factors affecting its accuracy and effective measures to avoid interference. High background microplastic contamination in typical lab waters (up to 1115 MP/L), glass fiber filter membrane and glassware were identified as dominant factors affecting microplastic quantification. The background microplastics in lab waters mainly originated from the process of water production and storage. A simple filtration process removed 99% of the background microplastic in the lab waters. After burning at 500 °C for 1 h, the microplastic contamination in the filter membrane and glassware was completely eliminated. H2O2 pretreatment and exposure time caused erroneous microplastic size assessment, and were suggested to be set at 48 h and 10 ms, respectively. During the extraction process, the residue in beakers reached ~ 20% and > 50% for 5 µm and 20 µm sized microplastics, respectively, greatly contributing to the microplastic loss. The comprehensive modified measures caused microplastic concentrations in the three typical samples detected by Nile Red/DAPI co-staining method to decrease by 65.7 - 92.2% and to approach the micro-Raman results. This study clarified the reasons for interfering with quantitative microplastics by fluorescent staining and the effective measures to avoid interference, which were conducive to improving the accuracy of quantitative methods of microplastics.

Role of suspended solids on the co-precipitation of pathogenic indicators and antibiotic resistance genes with struvite from digested swine wastewater.

Struvite recovered from wastewater contains high concentration of fecal indicator bacteria (FIB), porcine adenoviruses (PAdV) and antibiotic resistance genes (ARGs), becoming potential resources of these microbial hazards. Understanding the precipitation behavior of pathogenic indicators and ARGs with suspended solids (SS) will provide the possible strategy for the control of co-precipitation. In this study, SS was divided into high-density SS (separated by centrifugation) and low-density SS (further separated by filtration), and the role of SS on the co-precipitation of FIB, PAdV and ARGs was investigated. The distribution analysis showed that 35.5-73.0% FIB, 79.6% PAdV and 64.5-94.8% ARGs existed in high-density SS, while the corresponding values were 26.9-64.4%, 11.7% and 3.5-24.3% in low-density SS. During struvite generation, 82.7-96.9% FIB, 75.5% PAdV and 56.3-86.5% ARGs were co-precipitated into struvite. High-density SS contributed 20.7-68.5% FIB, 63.9% PAdV and 38.7-87.2% ARGs co-precipitation, and the corresponding contribution of low-density SS was 31.4-79.2%, 3.9% and 6.2-54.7%. Moreover, the precipitated SS in struvite obviously decreased inactivation efficiency of FIB and ARGs in drying process. These results provide a potential way to control the co-precipitation and inactivation of FIB, PAdV and ARGs in struvite through removing high-density SS prior to struvite recovery.

Modification of TiO2 by Er3+ and rGO enhancing visible photocatalytic degradation of arsanilic acid.

As a typical wide band gap photocatalyst, titania (TiO2) cannot use the visible light and has fast recombination rate of photogenerated electron-hole pairs. Simultaneous introduction of erbium ion (Er3+) and graphene oxide (rGO) into TiO2 might overcome these two drawbacks. In this study, Er3+ and rGO were co-doped on TiO2 to synthesize Er3+-rGO/TiO2 photocatalyst through a two-step sol-gel method. Based on the UV-visible diffuse reflectance spectra and photoluminescence spectrum, the introduction of Er3+ and rGO increased the visible light absorption efficiency and enhanced the migration of photogenerated electron. Pure TiO2 has almost no photocatalytic activity for arsanilic acid (p-ASA) degradation under visible light irradiation. However, while doping with 2.0 mol% Er3+ and 10.0 mol% rGO, the p-ASA could be completely degraded within 50 min by the Er3+-rGO/TiO2 photocatalyst under visible light irradiation, and most of produced inorganic arsenic was in situ removed by adsorption from the solution. The reactive oxygen species (ROS) reacting with p-ASA was determined and superoxide radical (O2•-) and singlet oxygen (1O2) were the dominant ROS for the oxidation of p-ASA and arsenite. This work provides an approach of introducing Er3+ and rGO to enhance the visible light photocatalytic efficiency of TiO2.

Co-precipitation of heavy metals with struvite from digested swine wastewater: Role of suspended solids.

Struvite production can recover ammonia and phosphorous from digested wastewater as fertilizer. During struvite generation, most of the heavy metals was co-precipitated with ammonia and phosphorous into struvite. Understanding the precipitation behavior of heavy metals with suspended solids (SS) might provide the possible strategy for the control of co-precipitation. In this study, the distribution of heavy metals in SS and their role on the co-precipitation during struvite recovery from digested swine wastewater were investigated. The results showed that the concentration of heavy metal (including Mn, Zn, Cu, Ni, Cr, Pb and As) ranged from 0.05 to 17.05 mg/L in the digested swine wastewater. The distribution analysis showed that SS with particles > 50 µm harbored most of individual heavy metal (41.3-55.6%), followed by particles 0.45-50 µm (20.9-43.3%), and SS-removed filtrate (5.2-32.9%). During struvite generation, 56.9-80.3% of individual heavy metal was co-precipitated into struvite. The contributions of SS with particles > 50 µm, 0.45-50 µm, and SS-removed filtrate on the individual heavy metal co-precipitation were 40.9-64.3%, 25.3-48.3% and 1.9-22.9%, respectively. These finding provides potential way for controlling the co-precipitation of heavy metals in struvite.

Photodegradation of roxarsone in the aquatic environment: influencing factors, mechanisms, and artificial neural network modeling.

Roxarsone (ROX), an organoarsenic feed additive, can be discharged into aquatic environment and photodegraded into more toxic inorganic arsenics. However, the photodegradation behavior of ROX in aquatic environment is still unclear. To better understand ROX photodegradation behavior, the influencing factors, photodegradation mechanism, and process modelling of ROX photodegradation were investigated in this study. The results showed that ROX in the aquatic environment was degraded to inorganic As(III) and As(V) under light irradiation. The degradation efficiency was enhanced by 25% with the increase of light intensity from 300 to 800 µW/cm2 via indirect photolysis. The photodegradation was temperature dependence, but was only slightly affected by pH. Nitrate ion (NO3-) had an obvious influence, but sulfate, carbonate, and chlorate ions had a negligible effect on ROX degradation. Dissolved organic matter (DOM) in the solution inhibited the photodegradation. ROX photodegradation was mainly mediated by reactive oxygen species (in the form of single oxygen 1O2) generated through ROX self-sensitization under irradiation. Based on the data of factors affecting ROX photodegradation, ROX photodegradation model was built and trained by an artificial neural network (ANN), and the predicted degradation rate was in good agreement with the real values with a root mean square error of 1.008. This study improved the understanding of ROX photodegradation behavior and provided a basis for controlling the pollution from ROX photodegradation.

Thermal crosslinking synthesis of ethylene-vinyl acetate copolymer supported floating TiO2 photocatalyst: characterization and photocatalytic performance.

Floating photocatalyst is of extensive interest due to easy recovery and efficient light harvest. Support materials largely determine the stability of floating photocatalysts and their synthesis complexity. Thus, finding proper floating supports is very important. Herein, ethylene-vinyl acetate copolymer (EVA) was investigated as a support to prepare floating TiO2/EVA using a simple thermal crosslinking procedure. Multiple characterization analyses demonstrated that TiO2 was anchored onto EVA surface evenly via hydrogen-bond-enhanced physical crosslinking and remained its virgin crystal structure. Photocatalytic experiments showed that the removal efficiency of Rhodamine B (RhB) by floating TiO2/EVA increased by 33.8% as compared to suspended particle TiO2. The h+ and ·O2- played dominant roles in TiO2/EVA-driven RhB degradation. A 30-day stability test demonstrated that TiO2/EVA had a high thermal, pH, and photo- stability. The three-run reuse test proved that TiO2/EVA exhibited satisfactory reusability. This study provides a new option for floating photocatalyst synthesis.

Zero-valent iron mediated alleviation of methanogenesis inhibition induced by organoarsenic roxarsone.

Zero-valent iron (ZVI) can enhance anaerobic digestion, and has great potential to alleviate/eliminate methanogenesis inhibition. Little is known about the feasibility of utilizing ZVI to alleviate methanogenesis inhibition that is caused by typical animal feed additive roxarsone in livestock wastewater. In this study, the role of ZVI on alleviating roxarsone-induced methanogenic inhibition and its mechanisms were investigated. With the increase of roxarsone concentration from 5 to 50 mg/L, the inhibition of methanogenesis increased from 3.0% to 65.7%. This inhibition was alleviated by 80.7% and 57.2% when 1.0 and 10.0 g/L ZVI were added, respectively. Due to ZVI addition, an efficient arsenic immobilization onto ZVI (45.4-85.8%) was achieved mainly through the formation of FeAsO4 precipitate and adsorption by ZVI. Under the function of ZVI, hydrogenotrophic methanogenic activity was obviously restored. The microbial community analysis indicates that the ZVI-regulated alleviation on the methanogenesis inhibition was attributed to the enrichment of Methanobacterium and Methanosarcina. The findings from this study demonstrate that ZVI addition is an effective way for treatment of organoarsenic-contaminated wastewater.

Response of anaerobic granular sludge to long-term loading of roxarsone: From macro- to micro-scale perspective.

Extensive use of organoarsenic feed additives such as roxarsone has caused organoarsenicals to occur in livestock wastewater and further within anaerobic wastewater treatment systems. Currently, information on the long-term impacts of roxarsone on anaerobic granular sludge (AGS) activity and the underlying mechanisms is very limited. In this study, the response of AGS to long-term loading of roxarsone was investigated using a laboratory up-flow anaerobic sludge blanket reactor spiked with 5.0 mg L-1 of roxarsone. Under the effect of roxarsone, methane production decreased by ∼40% due to the complete inhibition on acetoclastic methanogenic activity on day 260, before being restored eventually. Over 30% of the influent arsenic was accumulated in the AGS and the capability of AGS to prevent intracellular As(III) accumulation increased with time. The AGS size was reduced by ∼30% to 1.20‒1.26 mm. Based on morphology and confocal laser scanning microscopy analysis, roxarsone exposure stimulated the excretion of extracellular polymeric substances and the surface spalling of AGS. High-throughput sequencing analysis further indicated roxarsone initially altered the acidogenic pathway and severely inhibited the acetoclastic methanogen Methanothrix. Acetogenic bacteria and Methanothrix were finally enriched and became the main contributor for a full restoration of the initial methane production. These findings provide a deeper understanding on the effect of organoarsenicals on AGS, which is highly beneficial for the effective anaerobic treatment of organoarsenic-bearing wastewater.

Organoarsenic feed additives in biological wastewater treatment processes: Removal, biotransformation, and associated impacts.

Aromatic organoarsenicals are widely used in animal feeding operations and cause arsenic contamination on livestock wastewater and manure, thereby raising the risk of surface water pollution. Biological wastewater treatment processes are often used for livestock wastewater treatment. Organoarsenic removal and biotransformation under aerobic and anaerobic conditions, and the associated impacts have received extensive attention due to the potential threat to water security. The removal efficiency and biotransformation of organoarsenicals in biological treatment processes are reviewed. The underlying mechanisms are discussed in terms of functional microorganisms and genes. The impacts associated with organoarsenicals and their degradation products on microbial activity and performance of bioreactors are also documented. Based on the current research advancement, knowledge gaps and potential research in this field are discussed. Overall, this work delivers a comprehensive understanding on organoarsenic behaviors in biological wastewater treatment processes, and provides valuable information on the control of arsenic contamination from the degradation of organoarsenicals in biological wastewater treatment processes.

Inhibitory effect of oil and fat on denitrification using food waste fermentation liquid as carbon source.

Food waste fermentation liquid (FWFL) can be used as carbon source to enhance nitrogen removal in wastewater treatment. However, the influence of lipid, a common component of food waste, on denitrification remains unclear. In this study, the effect of oil and fat on denitrification process and the underlying mechanisms were investigated using synthetic oil- and fat-bearing carbon source and verified with real FWFL. In the batch experiment, oil and fat had no obvious influence on denitrification, but in the semi-continuous experiment, the denitrification rate in the oil- and fat-added assays decreased to 44% and 38% of that in the control, respectively, after 45 batches. Oil and fat caused sludge floatation, and the floating sludge thickness increased with the continuous operation. Oil/fat-sludge aggregates were observed in the floating sludge and limited gas release. Microbial community analysis indicated that oil and fat did not affect denitrifying bacteria abundance. Limitation of mass transfer might be the main reason for the inhibition of oil and fat on denitrification. In the real FWFL experiment, the denitrification rate in the original and emulsified oil-bearing FWFL decreased to 24% and 56% of that in the demulsifying FWFL, respectively, after 45 batches. These findings indicate the necessity of removing lipids when FWFL is used as denitrification carbon source.

Effect of ultrasonic and ozone pretreatment on the fate of enteric indicator bacteria and antibiotic resistance genes, and anaerobic digestion of dairy wastewater.

In this study, the effect of ultrasound (US), ozone and US combined with ozone (US/ozone) pretreatments on the fate of enteric indicator bacteria and antibiotic resistance genes (ARGs), and anaerobic digestion (AD) of dairy wastewater was investigated. The pretreatment conditions included US power 200 W, ozone concentration 4.2 mg O3/L, and pretreatment time 0-30 min. The results showed that US/ozone pretreatment was effective in the inactivation of enteric indicator bacteria. Total coliforms and enterococci were reduced by 99% and 92% after 30 min US/ozone pretreatment. Pretreatments could not decrease ARGs in absolute concentration, but could decrease ARGs in relative abundance. In the subsequent AD process, methane production increased more than 10% with 20 min ozone or 20 min US/ozone pretreatments. Pretreatment-AD together obviously inhibited the enrichment of ARGs in relative abundance. This study provided a pretreatment way to enhance methane production and to prevent the enrichment of ARGs.

Anaerobic biotransformation of roxarsone regulated by sulfate: Degradation, arsenic accumulation and volatilization.

Roxarsone, an extensively used organoarsenical feed additive, is often pooled in livestock wastewater. Sulfate exists ubiquitously in livestock wastewater and is capable for arsenic remediation. However, little is known about impacts of sulfate on roxarsone biotransformation during anaerobic digestion of livestock wastewater. In this study, the biodegradation of 5.0 mg L-1 roxarsone, and the accumulation and volatilization of the generated arsenical metabolites in a sulfate-spiked upflow anaerobic granular blanket reactor were investigated. Based on the analysis of degradation products, the nitro and arsenate groups of roxarsone were successively reduced to amino and arsenite groups before the C-As bond cleavage. Effluent arsenic concentration was ∼0.75 mg L-1, of which 82.9-98.5% were organoarsenicals. The maximum arsenic volatilization rate reached 32.6 µg-As kg-1-VS d-1. Adding 5.0 mg L-1 sulfate enabled 66.7% and 45.9% decrease in inorganic arsenic concentration and arsenic volatilization rate, respectively. Arsenic content in the anaerobic granular sludge (AGS) was accumulated to 1250 mg kg-1 within 420 days. Based on the results of FESEM-EDS and XPS, sulfate addition induced arsenic precipitation in the AGS through the formation of orpiment. Arsenic in the effluent, biogas and AGS accounted for 52.9%, 0.01% and 47.1% of the influent arsenic when the reactor operated stably. The findings from this study suggest that sulfate has effectively regulatory effects on arsenic immobilization and volatilization during anaerobic digestion of organoarsenic-contaminated livestock wastewater.

Impact of total solids content on anaerobic co-digestion of pig manure and food waste: Insights into shifting of the methanogenic pathway.

Dry anaerobic digestion (AD) has advantages over wet AD in treating high-solid organic wastes like livestock and food wastes, but an elevated total solids (TS) content would affect the AD performances. In this study, methane production of digesters co-digesting pig manure (PM) and food waste (FW) at different TS contents (R1, TS 5%; R2, TS 10%; R3, TS 15%; and R4, TS 20%) was assessed. The results showed the specific methane yield had no significant difference with the increase of TS contents from 5% to 15% (278.8-291.7 NmL/g VSadded), while it was reduced at a 20% TS content (259.8 NmL/g VSadded). Two peaks of total volatile fatty acids and daily methane production were observed in the high-solid digesters (R2-R4), while only one peak occurred in wet AD (R1). A new kinetics model was developed to describe the two-peak methane production behavior at high TS contents. The analysis on the microbial community structure clearly showed the different evolutions of methanogenic pathways in low and high solids content systems. In dry AD (R4), there was a general shifting from the acetoclastic pathway, to mixotrophic pathway and hydrogenotrophic pathway, with the dominance of mixotrophic and hydrogenotrophic methanogens.

Simultaneous roxarsone photocatalytic degradation and arsenic adsorption removal by TiO2/FeOOH hybrid.

Roxarsone (3-nitro-4-hydroxyphenylarsonic acid) is an extensively used organoarsenic feed additive. The effective removal of arsenic from roxarsone degradation before discharging is of great importance for controlling artificial arsenic pollution in aquatic environment. In this study, a bifunctional TiO2/ferrihydrite (TiO2/FeOOH) hybrid was synthesized by a hydrothermal method for the simultaneously photocatalytic degradation of roxarsone and adsorption removal of released arsenic. The analysis of the prepared TiO2/FeOOH by field-emission scanning electron microscope (FE-SEM), transmission electron microscopy (TEM), Raman spectra, X-ray diffraction (XRD), diffuse reflectance spectroscopy (DRS), and X-ray photoelectron spectroscopy (XPS) confirmed the successful formation of the hybrid of crystalline TiO2 and no crystalline FeOOH. TiO2/FeOOH hybrid had better adsorption capacity for As(V) than roxarsone. Compared to TiO2, the TiO2/FeOOH hybrid exhibited much superior UV-driven photocatalytic activities for roxarsone degradation. After 12 h irradiation, more than 96% of roxarsone was degraded by 1:1 TiO2/FeOOH hybrid, and the released As(V) was simultaneously removed from the solution. The residual As(V) concentration was lower than 0.02 mg L-1. The reusability test indicated that TiO2/FeOOH hybrid had excellent stability and reliability. The possible mechanism of roxarsone degradation and released inorganic arsenics removal by this hybrid was also proposed. These results clearly indicated that the TiO2/FeOOH hybrid could be used for the removal of roxarsone and its degradation product.

Efficiency of sequential UV/H2O2 and biofilm process for the treatment of secondary effluent.

In response to the shortage of water resources, multiple processes have been applied to turn wastewater secondary effluent (SE) into potable water. However, trace organic contaminants (TOrCs) and high concentrations of organic matter contained in SE pose a significant challenge to the reclamation. In this manuscript, combined UV-based and biofilm processes were used to treat the SE spiked with ibuprofen (IBU) and clofibric acid (CA). The efficiency of these sequential treatments was characterized in terms of changes in dissolved organic carbon (DOC), absorbance at 254 nm (A254), fluorescence excitation-emission matrix (FEEM), the concentration of IBU and CA, and molecular weight of SE. Parallel factor (PARAFAC) was applied as the analysis method for FEEM of the samples and two fluorescent components were successfully identified: humic-like substances (C1) and protein-like matter (C2). Large reductions in A254, C1, C2, IBU, and CA were observed during the UV-based processes, especially with the addition of H2O2. Nearly 50% of A254, 80% of the component C1 were decreased and almost complete removal of the component C2 and TOrCs was achieved by UV/2.0 mM H2O2 after 90-min treatment. During the oxidation processes, the formation of lower molecular weight (LMW) compounds was detected, and the biodegradability of the organic matters was greatly increased. Although no significant DOC reduction was obtained in UV-based processes, an obvious further DOC reduction (30~60%) was achieved by biofilm treatment following UV-based processes, especially after UV/H2O2 treatments. In the meantime, large amounts of LMW were removed in the biofilm treatment process. This manuscript provides an effective advanced treatment of SE for the removal of DOC and TOrCs, facilitating the wastewater reclamation.

Surfactant-enhanced anaerobic acidogenesis of Canna indica L. by rumen cultures.

Polyoxyethylene sorbitan monoolate (Tween 80) was used to enhance the anaerobic acidogenesis of Canna indica L. (canna) by rumen culture in this study. Dose of Tween 80 at 1 ml/l enhanced the volatile fatty acids (VFA) production from the acidogenesis of canna compared to the control. However, Tween 80 at higher dosages than 5 ml/l inhibited the rumen microbial activity and reduced the VFA yield. Response surface methodology was successfully used to optimize the VFA yield. A maximum of VFA yield of 0.147 g/g total solids (TS) added was obtained at canna and Tween 80 concentrations of 6.3g TS/l and 2.0 ml/l, respectively. Dosage of Tween 80 at 1-3.75 ml/l reduced the unproductive adsorption of microbes or enzymes on the lignin part in canna and increased microbial activity. A high VFA production was achieved from canna presoaked with Tween 80, suggesting that the structure of canna was disrupted by Tween 80.

Selection of seeding strategy for fast start-up of Anammox process with low concentration of Anammox sludge inoculum.

The long start-up time and large demand of Anammox seed sludge limit the practical application of Anammox process. In this study, the seeding strategy of fast start-up of Anammox process using 2.0 g VSS L-1 of anaerobic granular sludge (AGS) or activated sludge (AS) with various low concentration of Anammox sludge as inoculum was investigated. In laboratory scale, the start-up (achieving 70% TN removal) was shortened from 21 days to 5 days when Anammox sludge concentration increased from 0.02 g VSS L-1 to 0.2 g VSS L-1 with 2 g VSS L-1 AS as inoculum, and 16S rDNA analysis indicated the enrichment of Anammox bacteria, while the start-up failed with AGS. In pilot scale, the start-up was achieved in 10 days using 0.02 g VSS L-1 of Anammox sludge and 2.0 g VSS L-1 of AS, confirming the fast start-up of Anammox process with low concentration of Anammox sludge.