Spectroscopic evidence for hyperthermophilic pretreatment intensifying humification during pig manure and rice straw composting.

Hyperthermophilic pretreatment composting (HPC) is superior to traditional composting (CK) with shortened maturity period and enhanced humification degree. However, the chemical and structural evolution of humic substances (HS) at the molecular level is not known. In this study, the impact of hyperthermophilic pretreatment (90 °C, 4 h) on the content and chemical composition of HS during composting were investigated. The HS content of the final compost was 87.8 g/kg and 76.7 g/kg in HPC and CK, respectively. Significantly higher humic acid/fulvic acid ratio (1.27 in HPC v.s. 0.77 in CK) was observed in HPC. 13C NMR spectroscopic data showed a higher aromatics percentage and earlier enrichment of aromatic structures in HS extracted from HPC than CK. Intensified humification of HPC was related to the increased levels of HS precursors and degradation of lignocellulose. Redundancy analysis demonstrated that aromatic C, phenolic C and O-alkyl C can be used for evaluation of the humification degree.

Production and Characterization of Keratinolytic Proteases by a Chicken Feather-Degrading Thermophilic Strain, Thermoactinomyces sp. YT06.

Thermoactinomyces sp. strain YT06 was isolated from poultry compost and observed to degrade integral chicken feathers completely at 60°C, resulting in the formation of 3.24 mg/ml of free amino acids from 50 ml of culture containing 10 g/l chicken feathers. Strain YT06 could grow and secrete keratinase using feather as the only carbon and nitrogen sources without other supplement, but complementation of 10 g/l sucrose and 4 g/l NaNO3 increased the production of the keratinolytic enzyme. The maximum protease activity obtained was 110 U/ml and for keratinase was 42 U/ml. The keratinase maintained active status over a broad pH (pH 8-11) and temperature (60-75°C). It was inhibited by serine protease inhibitors and most metal ions; however, it could be stimulated by Mn²âº and the surfactant Tween-20. A reductive agent (ß-mercaptoethanol) was observed to cleave the disulfide bond of keratin and improve the access of the enzyme to the keratinaceous substrate. Zymogram analysis showed that strain YT06 primarily secreted keratinase with a molecular mass of approximately 35 kDa. The active band was assessed by MALDI-TOF mass spectrometry and was observed to be completely identical to an alkaline serine protease from Thermoactinomyces sp. Gus2-1. Thermoactinomyces sp. strain YT06 shows great potential as a novel candidate in enzymatic processing of hard-to-degrade proteins into high-value products, such as keratinous wastes.

Biochar decreases nitrogen oxide and enhances methane emissions via altering microbial community composition of anaerobic paddy soil.

Biochar application to agricultural soil is an appealing approach to mitigate nitrous oxide (N2O) and methane (CH4) emissions. However, the underlying microbial mechanisms are unclear. In this study, a paddy soil slurry was incubated anaerobically for 14d with biochar amendments produced from rice straw at 300, 500, or 700°C (B300, B500, and B700) to study their influences on greenhouse gas emissions. Illumina sequencing was used to characterize shift of soil bacterial and archaeal community composition. After peaking at day 1, N2O emission then sharply decreased to low levels while CH4 started to emit at day 3 then continually increased with incubation. Compared to control soil (57.9mgkg-1 soil), B300, B500, and B700 amendments decreased N2O peak emission to 17.9, 1.28, and 0.59mgkg-1, mainly due to increased soil pH. In contrast, the amendments enhanced CH4 production from 58.2 to 93.4, 62.6, and 63.4mgkg-1 at day 14 due to increased soil dissolved organic carbon. Abundance of denitrifying bacteria (e.g., Bacilli, 7.07-13.6 vs. 16.9%) was reduced with biochar amendments, especially with B500 and B700, contributing to the decreased N2O emissions. However, larger pore size of B500 and B700 (surface area of 68.1 and 161m2g-1) than B300 (4.40m2g-1) favored electron transfer between bacteria and iron minerals, leading to increased abundance of iron-reducing bacteria, (e.g., Clostridia, 48.2-50.6 vs. 33.3%), which competed with methanogens to produce CH4, thereby leading to lower increase in CH4 emission. Biochar amendments with high pH and surface area might be effective to mitigate emission of both N2O and CH4 from paddy soil.

[Degradation Characteristics of Spectral Analysis of Straw Nursery Containers in Situ Soil Culture].

This paper aimed to reveal the degradation behavior of a new type of biodegradable containers. The biodegradable containers, which was made of modified soybean adhesive and straw, was processed in situ biodegradation under natural condition. The physicochemical property and microstructure of straw nursery containers treated and untreated were characterized with Cellulose Tester, Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscope - Energy Dispersive X-ray Spectroscopy (SEM-EDS), and Thermo-gravimetry Analysis (TGA), respectively. The results indicated that the content of cellulose, hemicellulose and lignin of treated specimen decreased to 21.43%, 21.41% and 9.54% from 29.03%, 30.44% and 12.52%, respectively, comparing with those of untreated straw nursery container. FTIR analysis revealed that the ester and fat bond have been ruptured, and the aromatic characteristic peak became weakened. SEM-EDS spectrum showed the microfibril chain in the container has been fragmentation, and the soybean adhesive was also degradation. The surface of container appeared oxidization degradation. TGA analysis showed that a large number of small molecules have been produced in the process of degradation and the thermo-stability of treated samples improved.

Biochar increases arsenic release from an anaerobic paddy soil due to enhanced microbial reduction of iron and arsenic.

Previous studies have shown that biochar enhances microbial reduction of iron (Fe) oxyhydroxide under anaerobic incubation. However, there is a lack of data on its influence on arsenic (As) release from As-contaminated paddy soils. In this study, paddy soil slurries (120 mg As kg-1) were incubated under anaerobic conditions for 60 days with and without the addition of biochar (3%, w/w) prepared from rice straw at 500 °C. Arsenic release, Fe reduction, and As fractionation were determined at 1, 10, 20, 30, and 60 d, while Illumina sequencing and real-time PCR were used to characterize changes in soil microbial community structure and As transformation function genes. During the first month of incubation, As released into soil solution increased sharply from 27.9 and 55.9 to 486 and 630 µg kg-1 in unamended and biochar amended slurries, with inorganic trivalent As (AsIII) being the dominant specie (52.7-91.0% of total As). Compared to unamended slurries, biochar addition increased As and ferrous ion (Fe2+) concentrations in soil solution but decreased soil As concentration in the amorphous Fe/Al oxide fraction (F3). Difference in released As between biochar and unamended treatments (ΔAs) increased with incubation time, showing strong linear relationships (R2 = 0.23-0.33) with ΔFe2+ and ΔF3, confirming increased As release due to enhanced Fe reduction. Biochar addition increased the abundance of Fe reducing bacteria such as Clostridum (27.3% vs. 22.7%), Bacillus (3.34% vs. 2.39%), and Caloramator (4.46% vs. 3.88%). In addition, copy numbers in biochar amended slurries of respiratory As reducing (arrA) and detoxifying reducing genes (arsC) increased 19.0 and 1.70 fold, suggesting microbial reduction of pentavalent As (AsV) adsorbed on Fe oxides to AsIII, further contributing to increased As release.

Distribution of sulfonamides in liquid and solid anaerobic digestates: effects of hydraulic retention time and swine manure to rice straw ratio.

The effects of hydraulic retention time (20 and 15 days) and swine manure to rice straw ratios on distribution of sulfonamides (SAs) in liquid and solid anaerobic digestates were studied using bench-scale completely stirred tank reactors at (37 ± 1) °C. Results showed that anaerobic digestion (AD) treatment exhibited a good removal effect on sulfadiazine (SDZ), sulfadimidine (SM2) and sulfachloropyridazine (SCP), especially at HRT = 20 days and co-digestion with swine manure and rice straw. The removal rates of SDZ and SM2 were more than 90%, but only 72.8% for SCP. The residual SAs were mainly remained in solid digestates, with residual rates ranging from 28.8% to 71.3%, 40.6% to 88.0, and 82.7% to 97.0% for SDZ, SM2 and SCP, respectively. Due to lower pKa and higher log K ow of SCP, its residue in solid digestates was far more than SDZ and SM2. Higher HRT and co-digestion could improve the degradation of SAs, which can also be put down to the occurrence of cometabolism of SAs and COD.

Anaerobic digestion in mesophilic and room temperature conditions: Digestion performance and soil-borne pathogen survival.

Tomato plant waste (TPW) was used as the feedstock of a batch anaerobic reactor to evaluate the effect of anaerobic digestion on Ralstonia solanacearum and Phytophthora capsici survival. Batch experiments were carried out for TS (total solid) concentrations of 2%, 4% and 6% respectively, at mesophilic (37±1°C) and room (20-25°C) temperatures. Results showed that higher digestion performance was achieved under mesophilic digestion temperature and lower TS concentration conditions. The biogas production ranged from 71 to 416L/kg VS (volatile solids). The inactivation of anaerobic digestion tended to increase as digestion performance improved. The maximum log copies reduction of R. solanacearum and P. capsici detected by quantitative PCR (polymerase chain reaction) were 3.80 and 4.08 respectively in reactors with 4% TS concentration at mesophilic temperatures. However, both in mesophilic and room temperature conditions, the lowest reduction of R. solanacearum was found in the reactors with 6% TS concentration, which possessed the highest VFA (volatile fatty acid) concentration. These findings indicated that simple accumulation of VFAs failed to restrain R. solanacearum effectively, although the VFAs were considered poisonous. P. capsici was nearly completely dead under all conditions. Based on the digestion performance and the pathogen survival rate, a model was established to evaluate the digestate biosafety.

Ultra-small Nd(3+)-doped nanoparticles as near-infrared luminescent biolabels of hemin in bacteria.

Near-infrared (NIR) luminescent Nd(3+)-doped nanoparticles (NPs) have attracted considerable attention in bioimaging and biodetection. Here, we demonstrate sub-6 nm NaGdF4:Nd(3+),Fe(3+) NPs as luminescent biolabels of hemin molecules that act as the exogenous electron carriers in microbial communities. Contrary to the severe quenching of the visible luminescence for either upconverting or downconverting NPs, the Nd(3+)-doped NPs show superior properties in avoiding the optical absorption of hemin within the UV and visible spectral regions. A detailed examination showed that the Nd(3+)-doped NPs exhibit no obvious toxic effects on the microbial communities and show scarce influence on the characteristics of labeled hemin molecules in enhancing the reducing power of the fermentation system. More importantly, by monitoring the NIR luminescence of Nd(3+)-doped NPs, the selective accumulation of exogenous electron carriers in bacteria that are lacking reducing power has been revealed for the first time. The application of Nd(3+)-doped NPs as biolabels in bacteria would provide new opportunities for further unravelling the role of exogenous electron carriers in anaerobic digestion.

Using contaminated plants involved in phytoremediation for anaerobic digestion.

This study investigated the anaerobic digestion capability of five plants and the effects of copper (Cu) and S,S'-ethylenediaminedisuccinic acid (EDDS, a chelator widely used in chelant-assisted phytoremediation) on biogas production to determine a feasible disposal method for plants used in remediation. The results showed that in addition to Phytolacca americana L., plants such as Zea mays L., Brassica napus L., Elsholtzia splendens Nakai ex F. Maekawa, and Oenothera biennis L. performed well in biogas production. Among these, O. biennis required the shortest period to finish anaerobic digestion. Compared to normal plants with low Cu content, the plants used in remediation with increased Cu levels (100 mg kg(-1)) not only promoted anaerobic digestion and required a shorter anaerobic digestion time, but also increased the methane content in biogas. When the Cu content in plants increased to 500, 1000, and 5000 mg kg(-1), the cumulative biogas production decreased by 12.3%, 14.6%, and 41.2%, respectively. Studies also found that EDDS conspicuously restrained biogas production from anaerobic digestion. The results suggest that anaerobic digestion has great potential for the disposal of contaminated plants and may provide a solution for the resource utilization of plants used in remediation.

Methane production from wheat straw with anaerobic sludge by heme supplementation.

Wheat straw particles were directly used as substrate for batch anaerobic digestion with anaerobic sludge under 35°C to evaluate the effects of adding heme on methane production. When 1mg/l heme was added to the fermentation process with no agitated speed, a maximum cumulative methane production of 12227.8ml was obtained with cumulative methane yield of wheat straw was 257.4ml/g-TS (total solid), which was increased by 20.6% compared with 213.5ml/g-TS of no heme was added in the reactor. Meanwhile, oxido-reduction potential (ORP) level was decreased, the activity of coenzyme F420 was significantly improved and NADH/NAD(+) ratio were the highest than other experimental groups. These results suggest that heme-supplemented anaerobic sludge with no agitated speed may be providing a more reductive environment, which is a cost-effective method of anaerobic digestion from biomass waste to produce methane with less energy consuming.

Biochar pyrolytically produced from municipal solid wastes for aqueous As(V) removal: adsorption property and its improvement with KOH activation.

Biochar converted from waste products is being considered as an alternative adsorbent for removal of aqueous heavy metal(loid)s. In this work, experimental and modeling investigations were conducted to examine the effect of biochars pyrolytically produced from municipal solid wastes on removing aqueous As(V) before and after activated by 2M KOH solution. Results showed that the highest adsorption capacity of pristine biochars was 24.49 mg/g. The pseudo-second-order model and Langmuir adsorption isotherm model can preferably describe the adsorption process. The activated biochar showed enhanced As(V) adsorption ability with an adsorption capacity of 30.98 mg/g, which was more than 1.3 times of pristine biochars, and 2-10 times of modified biochars reported by other literatures. Increase of surface area and changes of porous texture, especially the functional groups on the surface of activated biochars are the major contributors to its more efficient adsorption of As(V).

Effect of biofumigation and chemical fumigation on soil microbial community structure and control of pepper Phytophthora blight.

Soil biofumigation with brassica plant residues has been shown to significantly suppress soilborne pathogen. However, little published data reported the impact of biofumigation on microbial community structure in pepper (Capsicum annuum L.) production systems under field conditions. Biofumigation with rapeseed (Brassica napus 'Dwarf Essex') meal and chemical fumigation with dazomet were tested to control the pepper disease caused by Phytophthora capsici. BF treatment showed the lowest disease incidence among these treatments. Effects on soil bacterial and fungal communities were assessed by denaturating gradient gel electrophoresis and the results showed that the biofumigation increased bacterial diversity and decreased fungal diversity. There was a negative correlation between soil bacterial diversity and disease incidence and a positive correlation between soil fungal diversity and disease incidence. Cloning of the microbial community showed that the microbial community structures were altered by biofumigation. Soil was also evaluated for their chemical properties. Biofumigation increased soil content of total N, NO3(-)-N, available P and available K. A significant correlation between soil microbial community structures and soil chemical properties was found. Overall, these results indicated that biofumigation reduced disease incidence of pepper through altering soil microbial community structures.

[Effect of pretreatment on storage and biogas production of baling wheat straw].

Long-term storage of crop straw is very important for biogas plant while pretreatment is always used to improve biogas production of crop straw. Feasibility of integrating the storage with pretreatment of baling wheat straw was studied. Changes of physicochemical properties and the biogas productivity of wheat straw obtained before and after 120 days storage were analyzed. The results showed that it was feasible to directly bale wheat straw for storage (control) and storage treatment had little effect on the physicochemical properties, structure and biogas productivity of wheat straw. After 120 day's storage, biogas production potential of the surface wheat straw of pile was decreased by 7.40%. Integrating NaOH pretreatment with straw storage was good for biogas production of wheat straw and the total solid (TS) biogas yield was increased by 7.02%-8.31% (compared to that of wheat straw without storage) and 5.68% -16.96% (compared to that of storage without alkaline pretreatment), respectively. Storage with urea treatment was adverse to biogas production of wheat straw and the contents of cellulose and hemicellulose of wheat straw were decreased by 18.25%-27.22% and 5.31%-16.15% and the TS biogas yield was decreased by 2.80%-7.71% after 120 day's storage. Exposing wheat straw to the air during the storage process was adverse to the conserving of organic matter and biogas utilization of wheat straw, but the influence was very slight and the TS biogas yield of wheat straw obtained from pile surface of control and urea treatment was decreased by 7.40% and 4.25%, respectively.

The fate of antagonistic microorganisms and antimicrobial substances during anaerobic digestion of pig and dairy manure.

The goals of the present study were to evaluate the suppressive capability of anaerobically digested slurry (ADS) against Phytophthora capsici and to determine the key factors of disease control in ADS. This was achieved by the investigations of the changes in microbial populations and the levels of antimicrobial compound during anaerobic digestion (AD). AD had no significant impact on the numbers of antagonistic fluorescent pseudomonads or Bacillus sp. The contents of total phenolics, volatile fatty acids and sugar fed with the raw slurries to the reactors were decreased by AD. However, the bioreactor effluents had higher concentrations of humic substances and ammonia than the feedstocks. Moreover, AD had a different influence on the content of amino acid in the pig manure compared to the dairy manure. The results obtained indicated that the key inhibitory factors of ADS might be attributed to ammonia and humic substances.

[Methane production by anaerobic co-digestion of chicken manure and Spartina alterniflora residue after producing methane].

The characteristics of Spartina alterniflora residue after producing methane (SAR) digested alone and co-digestion of various proportions of chicken manure and SAR were investigated by batch model at 35 degrees C +/- 1 degrees C. The initial total solid (TS) loading of all treatments was 8.0%. The results indicated that there was still some biogas produced by SAR with TS biogas yield of 107.25 mL x g(-1) and average methane content of 76.92%. The cellulose crystallinity of re-digested SAR was destructed by anaerobic microorganisms, and the destruction rate was 5.55%. Co-digestion meliorated the environment where microorganisms lived in and increased the cumulative biogas yield. When the ratio of chicken manure to SAR was 4 : 1, the cumulative biogas yield was highest while the cumulative biogas yield of T1 (the rate of chicken manure to SAR is 5 : 0 based on TS), T3-T6 (the rate of chicken manure to SAR are 3 : 2, 2 : 3, 1 :4 and 0 : 5 based on TS, respectively) were 61.31%, 62.09%, 52.15%, 39.74% and 31.67% of it. The anaerobic fermentation type of co-digestion is mix acid type fermentation. Co-digested with chicken manure promoted the destruction of cellulous crystallinity by 1.13% - 21.61% and especially when the rate of chicken manure to SAR was 4 : 1.

[Effect of alkaline post-treatment on physicochemical property of digested Spartina alterniflora].

Alkaline treatment is widely used for improving biogas production for lignocellulosic materials. This study was conducted to investigate the effect of alkaline treatment on physicochemical property of digested Spartina alterniflora (DSA). Fourier Transform Infrared Spectroscopy (FTIR), X-Ray diffraction patterns, proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR), solid-state 13C-NMR Spectroscopy and some general indicators were used to analyze changes of the lignocellulosic structure and composition of NaOH-treated digested Spartina alterniflora. The results showed that, after NaOH treatment, surface lignin and some carbohydrate were destructed into lignin fragment, organic acids and some other small molecular organic matter, but the skeleton structure of lignin and cellulose of DSA were not destructed significantly. The crystalline of cellulose of DSA was transferred into biodegradable forms and content of crystalline of DSA and cellulose were increased after NaOH treatment. The results of 13C-NMR showed that methyl (CH3) and carboxylic C(COOH) groups of DSA were decomposed significantly.

[Effect of NaOH-treatment on dry-thermophilic anaerobic digestion of Spartina alterniflora].

In order to improve the biotransformation rate of lignocellulosic materials, sodium hydroxide (NaOH) was widely used to pretreat lignocellulosic materials. Effect of NaOH-treatment on dry-thermophilic anaerobic digestion of Spartina alternflora was studied by batch model under the temperature of 55 degrees C +/- 1 degrees C, at the initial total solid loading (TSL) of 20%. The results indicated that biogas production was inhibited by NaOH-treatment and improved by NaOH-treatment with water washed. The cumulative biogas yield of control (CK), NaOH-treated and NaOH-treated with water washed (NaOH + water) were 268.35 mL/g, 205.76 mL/g and 299.97 mL/g, respectively. The methane content of CK and NaOH + water treatments kept stable while fluctuation of NaOH-treated treatment during anaerobic digestion process was observed. Compared with CK and NaOH + water treatments, methane content of NaOH-treated treatment was improved by 5.30%. The content of hemi-cellulose and cellulose of S. alternifora decreased while content of lignin of S. alterniflora increased after 51-day anaerobic digestion. The crystallinity of cellulose of S. alterniflora increased after NaOH-treatment which was consistent to the result of FTIR. The lignocellulosic structure was destroyed and the biodegradability of S. alterniflora was increased by NaOH pretreatment. However, the amount of Na+ was taken into the anaerobic system, besides the high Na+ content in the plant itself which inhibited the anaerobic microorganisms. Therefore, NaOH-treatment is considered to be unsuitable for the anaerobic digestion of S. alterniflora.

[Effect of NaOH-treatment on advanced anaerobic biogasification of Spartina alterniflora].

In order to improve the biotransformation rate of Sparnina alterniflora, effect of NaOH-treatment on anaerobic dry-mesophilic digestion of Spartina alterniflora and feasibility of NaOH-treatment as a pretreatment of biogas residues of Spartina alterniflora for advanced anaerobic biogasification were conducted under lab-scale conditions. The results indicated that there was less improvement to biogas yield with NaOH-treatment and the cumulative biogas yield of Spartina alterniflora was 358.94 mL/g TS which was 92.42% to that of control (CK). However, the average methane content was improved slightly with 1.84% improvement. After solid-state pretreatment with 5% NaOH solution for 48 h, the biogas residue of Spartina alterniflora was used for advanced biogasification. This experiment was conducted under 35 degrees C +/- 1 degrees C with initial total solid loading of 8%. The cumulative biogas yield was 209.73 mL/g TS with 70.78% of average methane content, but the biotransformation rate was only 23.29% which was much lower than that of Spartina alterniflora. The fermentation type was propionic acid type fermentation. After two-phase fermentation treatment, cellulose content was decreased significantly while lignin and hemicellulose content were increased. The crystalinity of cellulose of biogas residue decreased after two-phase anaerobic fermentation which was consistent to result of FTIR. The comprehensive analysis of experiment indicated that biogas residue of Spartina alterniflora was still a good material for biogas production and NaOH-treatment was a good pretreatment for biogas production.

Distribution of heavy metal contents and chemical fractions in anaerobically digested manure slurry.

Digested slurry samples from twenty-one large-scale anaerobic digestion plants together with intensive pig and dairy farms in Jiangsu Province of China were collected and analyzed for total and dissolved concentrations of Zn, Cu and As, as well as chemical characteristics. The results showed that total concentrations of Zn, Cu and As in digested pig slurries were concentrated to <10, <5 and 0.02-0.1 mg/l, respectively; while <2 and 10-30, <1, and 0.02-0.1 mg/l, respectively, in digested dairy slurries. Lowering the dietary supply of these elements to pig and dairy would be the most effective way to control heavy metal contents in digested manure slurries. Dissolved fractions of Zn, Cu and As accounted for 1-74%, 1-33% and 2-53% of the total concentrations, respectively, in digested pig slurries; and 18-65%, 12-58% and 3-68% in digested dairy slurries. The chemical fractions of heavy metals in digested slurries were not only dependent on the total concentrations of heavy metals in raw manures but also on conditions of digestion and storage. Oxidation pond systems could significantly cripple the total contents of heavy metals in digested slurries, and the removal effect was better in multi-oxidation-pond systems than that in primary-oxidation-pond systems. However, the chemical fractions of heavy metals in digested slurries changed in a complicated manner when stored in oxidation ponds, due to the suspended solid deposition, elements reduction, as well as variations of pH values and oxidation-reduction potential.

Removal of nutrients and veterinary antibiotics from swine wastewater by a constructed macrophyte floating bed system.

The potential of three varieties of Italian ryegrass (Lolium multiflorum Lam.), Dryan, Tachimasari and Waseyutaka, to improve the water quality of swine wastewater was evaluated using a constructed macrophyte floating bed system. With respect to reductions in levels of nutrients, chemical oxygen demand (COD), and sulfonamide antimicrobials (SAs, including sulfadiazine, sulfamethazine, and sulfamethoxazole), Dryan performed better than Tachimasari and Waseyutaka. For Dryan, total N was reduced by 84.0%, total P by 90.4%, COD by 83.4% and sulfonamide antimicrobials by 91.8-99.5%. Similar results were observed for Tachimasari and Waseyutaka. The results indicated that the treatment of swine wastewater using the constructed macrophyte floating bed system was effective in the removal of nutrients and veterinary antibiotics.