Integrated nutrient recycling: Ammonia recovery from thermophilic composting of shrimp aquaculture sludge via self-heated bench-scale reactor and mango plant growth enhancement by the compost.

Due to the rapid growth of the aquaculture industry, large amounts of organic waste are released into nature and polluted the environment. Traditional organic waste treatment such as composting is a time-consuming process that retains the ammonia (NH3) in the compost, and the compost produced has little economic value as organic fertilizer. Illegal direct discharge into the environment is therefore widespread. This study investigates the recovery of NH3 through thermophilic composting of shrimp aquaculture sludge (SAS) and its application as a soil conditioner for the growth of mango plants. A maximum composting temperature of 57.10 °C was achieved through self-heating in a 200 L bench-scale reactor, resulting in NH3 recovery of 224.04 mol/ton-ds after 14 days. The addition of calcium hydroxide and increased aeration have been shown to increase NH3 volatilization. The recovered NH3 up to 3 kg-N can be used as a source of clean nitrogen for high-value microalgae cultivation, with a theoretical yield of up to 34.85 kg-algae of microalgae biomass from 1 ton-ds of SAS composting. Despite the high salinity, SAS compost improved mango plant growth and disease resistance. These results highlight the potential of SAS compost as a sustainable source of clean nitrogen for microalgae cultivation and soil conditioner, contributing to a waste-free circular economy through nutrient recycling and sustainable agriculture.

Enhancing algal growth and nutrient recovery from anaerobic digestion piggery effluent by an integrated pretreatment strategy of ammonia stripping and flocculation.

Anaerobic digestion piggery effluent (ADPE) with a quite high ammonium (NH4 +) concentration and turbidity (dark brown color) generally requires high dilution before microalgae cultivation, owing to its NH4 + toxicity and color inhibition to algal growth. An integrated pretreatment strategy of ammonia stripping and chemical flocculation may be a more practical pretreatment procedure for enhancing algae yield and nutrient recovery from anaerobic digestion piggery effluent. In this study, we determined the optimum pretreatment strategy of anaerobic digestion piggery effluent for subsequent microalgae cultivation and nutrient recovery. The results showed that the integrated anaerobic digestion piggery effluent pretreatment strategy of high-temperature ammonia stripping and chemical flocculation at a mixed dosage of 2 g L-1 polyaluminum chloride (PAC) and 40 mg L-1 cationic polyacrylamide (C-PAM), and 50 mg L-1 ammonium nitrogen (NH4 +-N) enrichment provided maximum algal yield (optical density = 1.8) and nutrient removal (95.2%, 98.7%, 99.3%, and 78.5% for the removal efficiencies of total nitrogen, NH4 +-N, total phosphorus, and chemical oxygen demand, respectively) from anaerobic digestion piggery effluent. The integrated pretreatment strategy is expected to become a more practical pretreatment procedure for enhancing algae yield and nutrient recovery from anaerobic digestion piggery effluent.

The effect of calcium hydroxide addition on enhancing ammonia recovery during thermophilic composting in a self-heated pilot-scale reactor.

A modified outdoor large-scale nutrient recycling system was developed to compost organic sludge and aimed to recover clean nitrogen for the cultivation of high-value-added microalgae. This study investigated the effect of calcium hydroxide addition on enhancing NH3 recovery in a pilot-scale reactor self-heated by metabolic heat of microorganisms during thermophilic composting of dewatered cow dung. 350 kg-ww of compost was prepared at the ratio of 5: 14: 1 (dewatered cowdung: rice husk: compost-seed) in a 4 m3 cylindrical rotary drum composting reactor for 14 days of aerated composting. High compost temperature up to 67 °C was observed from day 1 of composting, proving that thermophilic composting was achieved through the self-heating process. The temperature of compost increases as microbial activity increases and temperature decreases as organic matter decreases. The high CO2 evolution rate on day 0-2 (0.02-0.08 mol/min) indicated that microorganisms are most active in degrading organic matter. The increasing conversion of carbon demonstrated that organic carbon was degraded by microbial activity and emitted as CO2. The nitrogen mass balance revealed that adding calcium hydroxide to the compost and increasing the aeration rate on day 3 volatilized 9.83 % of the remaining ammonium ions in the compost, thereby improving the ammonia recovery. Moreover, Geobacillus was found to be the most dominant bacteria under elevated temperature that functions in the hydrolysis of non-dissolved nitrogen for better NH3 recovery. The presented results show that by thermophilic composting 1 ton-ds of dewatered cowdung for NH3 recovery, up to 11.54 kg-ds of microalgae can be produced.

Ingestion rate estimated from food concentration and predatory role of copepod nauplii in the microbial food web of temperate embayment waters.

To quantitatively evaluate the role of copepod nauplii as predators in the microbial food web, the ingestion rate (IR) of copepod nauplii and the food requirement (FR) of microzooplankton were estimated monthly for 3 consecutive years in temperate embayment waters. The IR of dominant copepod nauplii (Acartia spp. nauplii) was estimated from water temperature, individual carbon weight and food concentration and peaked (>0.50 µgC ind-1 d-1) with relatively high food concentration (>57.5 µgC L-1). This result suggests that food concentration should be considered to estimate copepod naupliar IR in marine environments, especially where biological conditions fluctuate largely. The comparison of copepod naupliar and microprotozoan FR showed the dominance of naked ciliate FR (77.0-90.2%) during the study period except in spring when comparable values were observed between the FR of naked ciliates (41.6%) and copepod nauplii (33.6%). During spring, transfer efficiency (10.5%) from primary production (PP) to microzooplankton production was lower than in other seasons (16.2-17.1%). This study indicates that copepod nauplii are seasonally important micro-sized predators in the microbial food web of temperate embayment waters and that carbon flow through copepod nauplii is a pathway which inefficiently transfers PP to higher trophic levels.

Microalgal flocculation and sedimentation: spatiotemporal evaluation of the effects of the pH and calcium concentration.

The high cost of harvesting microalgae is a major hurdle for the microalgae industry, and an efficient pre-concentration method is required. In this study, the effects of using different pH values (between pH 3 and 11) and calcium (Ca2+) concentrations (between 0 and 5 mM) on Chlorella vulgaris sedimentation were investigated by evaluating the spacio-temporal distributions of microalgae cells. Fast and efficient sedimentation occurred (within 10 min) at a high Ca2+ concentration (5 mM) at pH 9 and 11. However, the sediment volume was lower at a Ca2+ concentration of 3 mM than at a Ca2+ concentration of 5 mM. This indicated that the Ca2+ concentration strongly affected the sediment volume. Fast sedimentation and a low sediment volume were found at pH 7 and a Ca2+ concentration of 5 mM, probably because of the neutral charge in the system (adhesion to calcium precipitates would have occurred at a high pH). The highest Ca2+ recovery (82%) was achieved when sediment produced at pH 11 and a Ca2+ concentration of 5 mM was acidified to pH 3.

Advanced light-tolerant microalgae-nitrifying bacteria consortia for stable ammonia removal under strong light irradiation using light-shielding hydrogel.

The consortium of microalgae and nitrifying bacteria has attracted attention owing to its advantages, such as energy- and cost-efficiency in terms of using only light irradiation without aeration. However, high light intensity can easily cause photoinhibition of nitrifying bacteria, resulting in process breakdown of the consortium. This challenge limits its practical application in outdoor environment. In a previous study, we developed a "light-shielding hydrogel" which entrapped nitrifying bacteria in carbon black-added alginate hydrogel beads and confirmed its effectiveness of protecting the nitrifying bacteria from intense light up to 1600 µmol photons m-2 s-1. However, the applicability of the light-shielding hydrogel to microalgae-nitrifying bacteria consortia under strong light irradiation has not yet been clarified. In this study, we aimed to establish consortia of Chlorella sorokiniana and nitrifying bacteria immobilised in light-shielding hydrogel and evaluate their nitrification performance under strong light. Three nitrifying bacteria conditions were used: light-shielding hydrogel, hydrogel containing only nitrifying bacteria without carbon black ('hydrogel'), and dispersed nitrifier without immobilisation ('dispersion') as a control. At 1600 µmol photons m-2 s-1, the dispersion afforded a significant decrease in nitrification activity and subsequent process breakdown. In contrast, light-shielding hydrogel achieved complete nitrification without nitrite accumulation and had nitrification rates of approximately nine and two times higher than those for the dispersion and hydrogel conditions, respectively. Based on the overall evaluation, a possible sequence of process breakdown under strong light was also proposed. This study demonstrated for the first time that the light-shielding hydrogel/consortia combination had potential for applications, which require mitigation of photoinhibition under strong light irradiation. Further, it is expected that the proposed method will contribute to realise the practical application of microalgae-nitrifying bacteria consortia in various countries that experience high sunlight intensity due to their location in the sunbelt areas.

Inoculation of Neurospora sp. for improving ammonia production during thermophilic composting of organic sludge.

Recent attempts have been made to develop a thermophilic composting process for organic sludge to not only produce organic fertilizers and soil conditioners, but to also utilize the generated ammonia gas to produce high value-added algae. The hydrolysis of organic nitrogen in sludge is a bottleneck in ammonia conversion, and its improvement is a major challenge. The present study aimed to elucidate the effects of inoculated Neurospora sp. on organic matter decomposition and ammonia conversion during thermophilic composting of two organic sludge types: anaerobic digestion sludge and shrimp pond sludge. A laboratory-scale sludge composting experiment was conducted with a 6-day pretreatment period at 30 °C with Neurospora sp., followed by a 10-day thermophilic composting period at 50 °C by inoculating the bacterial community. The final organic matter decomposition was significantly higher in the sludge pretreated with Neurospora sp. than in the untreated sludge. Correspondingly, the amount of non-dissolved nitrogen was also markedly reduced by pretreatment, and the ammonia conversion rate was notably improved. Five enzymes exhibiting high activity only during the pretreatment period were identified, while no or low activity was observed during the subsequent thermophilic composting period, suggesting the involvement of these enzymes in the degradation of hardly degradable fractions, such as bacterial cells. The bacterial community analysis and its function prediction suggested the contribution of Bacillaceae in the degradation of easily degradable organic matter, but the entire bacterial community was highly incapable in degrading the hardly degradable fraction. To conclude, this study is the first to demonstrate that Neurospora sp. decomposes those organic nitrogen fractions that require a long time to be decomposed by the bacterial community during thermophilic composting.

Effect of enzymatic pre-treatment on thermophilic composting of shrimp pond sludge to improve ammonia recovery.

In recent years, attempts have been made to develop a thermophilic composting process for organic sludge to produce ammonia gas for high value-added algal production. However, the hydrolysis of non-dissolved organic nitrogen in sludge is a bottleneck for ammonia conversion. The aim of this study was to identify enzymes that enhance sludge hydrolysis in a thermophilic composting system for ammonia recovery from shrimp pond sludge. This was achieved by screening useful enzymes to degrade non-dissolved nitrogen and subsequently investigating their effectiveness in lab-scale composting systems. Among the four hydrolytic enzyme classes assessed (lysozyme, protease, phospholipase, and collagenase), proteases from Streptomyces griseus were the most effective at hydrolysing non-dissolved nitrogen in the sludge. After composting sludge pre-treated with proteases, the final amount of non-dissolved nitrogen was 46.2% of the total N in the control sample and 22.3% of the total N in the protease sample, thus increasing the ammonia (gaseous and in-compost) conversion efficiency from 41.5% to 56.4% of the total N. The decrease in non-dissolved nitrogen was greater in the protease sample than in the control sample during the pre-treatment period, and no difference was observed during the subsequent composting period. These results suggest that Streptomyces proteases hydrolyse the organic nitrogen fraction, which cannot be degraded by the bacterial community in the compost. Functional potential analysis of the bacterial community using PICRUSt2 suggested that 4 (EC:3.4.21.80, EC:3.4.21.81, EC:3.4.21.82, and EC:3.4.24.77) out of 13 endopeptidase genes in S. griseus were largely absent in the compost bacterial community and that they play a key role in the hydrolysis of non-dissolved nitrogen. This is the first study to identify the enzymes that enhance the hydrolysis of shrimp pond sludge and to show that the thermophilic bacterial community involved in composting has a low ability to secrete these enzymes.

Transcriptome-wide study in the green microalga Messastrum gracile SE-MC4 identifies prominent roles of photosynthetic integral membrane protein genes during exponential growth stage.

The non-model microalga Messastrum gracile SE-MC4 is a potential species for biodiesel production. However, low biomass productivity hinders it from passing the life cycle assessment for biodiesel production. Therefore, the current study was aimed at uncovering the differences in the transcriptome profiles of the microalgae at early exponential and early stationary growth phases and dissecting the roles of specific differential expressed genes (DEGs) involved in cell division during M. gracile cultivation. The transcriptome analysis revealed that the photosynthetic integral membrane protein genes such as photosynthetic antenna protein were severely down-regulated during the stationary growth phase. In addition, the signaling pathways involving transcription, glyoxylate metabolism and carbon metabolism were also down-regulated during stationary growth phase. Current findings suggested that the coordination between photosynthetic integral membrane protein genes, signaling through transcription and carbon metabolism classified as prominent strategies during exponential growth stage. These findings can be applied in genetic improvement of M. gracile for biodiesel application.

Mechanism of cell proliferation during starvation in a continuous stirred tank anaerobic reactor treating food waste.

Anaerobic digestion is a mature technology; however, the mechanism of cell proliferation during starvation has not yet been clarified. In this study, a continuous stirred tank reactor (CSTR) treating food waste was exposed to deliberate starvation for 12 days. The cell density and the variability of digestate characteristics during starvation were monitored. Starvation increased cell density from 2.8 × 1010 to 7.9 × 1010 cells mL-1 within 2 days and reduced the residual substrate. This increase in cell density was suggested owing to a switch of the anaerobic digester microorganisms' substrate preference to the complex fractions because the easily digestible fractions were exhausted. The prolonged starvation of more than approximately 3-6 days induced an increase in the free ammonia concentration to an inhibitive level of more than 0.10 g-N L-1 for anaerobic digestion microorganisms due to the excessive ammonification of residual nitrogen, thereby resulting in a drastic decrease in cell density. Our results demonstrated that a deliberate starvation operation in an appropriate timeframe applied to a CSTR treating food waste is beneficial to proliferate cells and, at the same time, reduce residual substrate.

Tolerance of Tetraselmis tetrathele to High Ammonium Nitrogen and Its Effect on Growth Rate, Carotenoid, and Fatty Acids Productivity.

Microalgae can use either ammonium or nitrate for its growth and vitality. However, at a certain level of concentration, ammonium nitrogen exhibits toxicity which consequently can inhibit microalgae productivity. Therefore, this study is aimed to investigate the tolerance of Tetraselmis tetrathele to high ammonium nitrogen concentrations and its effects on growth rate, photosynthetic efficiency (F v /F m ), pigment contents (chlorophyll a, lutein, neoxanthin, and ß-carotene), and fatty acids production. Experiments were performed at different ammonium nitrogen concentrations (0.31-0.87 gL-1) for 6 days under a light source with an intensity of 300 µmol photons m-2 s-1 and nitrate-nitrogen source as the experimental control. The findings indicated no apparent enhancement of photosynthetic efficiency (F v/F m) at high levels of ammonium nitrogen ( NH 4 + -N) for T. tetrathele within 24 h. However, after 24 h, the photosynthetic efficiency of T. tetrathele increased significantly (p < 0.05) in high concentration of NH 4 + -N. Chlorophyll a content in T. tetrathele grown in all of the different NH 4 + -N levels increased significantly compared to nitrate-nitrogen (NO3-N) treatment (p < 0.05); which supported that this microalgal could grow even in high level of NH 4 + -N concentrations. The findings also indicated that T. tetrathele is highly resistant to high ammonium nitrogen which suggests T. tetrathele to be used in the aquaculture industry for bioremediation purpose to remove ammonium nitrogen, thus reducing the production cost while improving the water quality.

Storage capacity for phosphorus during growth and maturation in a brown alga Sargassum macrocarpum.

Improved coastal management has decreased anthropogenic nutrient input over the past few decades, leading to phosphorus depletion. It has been hypothesized that phosphorus depletion in coastal environments leads to declines in macroalgae abundance. Perennial canopy-forming temperate macroalgae can experience the effects of limited phosphorus availability during seasonal phosphorus depletion periods. When nutrients are sufficient, they are stored in algal tissues after luxury uptake and are available to support growth during phosphorus-depleted conditions. Cultivation of mature and actively growing juvenile brown alga (Sargassum macrocarpum) under different nutrient conditions provided individuals with different tissue nutrient concentrations. The maximum photosynthetic rates of these individuals were examined under nutrient-depleted conditions to evaluate "storage capacity", which we defined as the amount of stored phosphorus that can support maximum growth. Maximum photosynthetic rate was used as a proxy for maximum growth rates. The experiments revealed that growth rates of juveniles increased when stored phosphorus content was high. In contrast, the maximum growth rates tended not to increase with an increase in stored phosphorus content in mature individuals. The phosphorus storage capacities for juvenile and mature individuals were approximately 19 and more than 16 weeks, respectively, suggesting that individual alga can endure several months of phosphorus depletion.

Nitrification of anaerobic digestate using a consortium of microalgae and nitrifiers in an open photobioreactor with moving bed carriers.

A consortium of microalgae and nitrifiers has attracted attention as an alternative to the expensive traditional nitrification process. A possible obstacle to achieving this is the inhibition of nitrifiers under strong light irradiation. This study evaluated the effect of moving bed carriers on anaerobic digestate nitrification in an open photobioreactor inoculated with microalgae and nitrifiers under an incident light intensity of 1000 µmol photons m-2 s-1. The results showed higher specific nitrification activity in the carrier-added photobioreactor (103.6 mg-N g-TSS day-1) than in one in which no carrier was added (11.7 mg-N g-TSS day-1). The empirical equations for determining the light intensity at different depths in the photobioreactor showed a significant contribution by carriers in attenuating the incident light intensity. This is due to the large light attenuation caused by the carrier (1.09 cm-1). The average light intensity inside of the photobioreactor decreased considerably in the carrier-added photobioreactor (342 µmol photons m-2 s-1), whereas it did not decrease in the one with no added carrier. It was found that specific nitrification activity was significantly negatively affected by average light intensity inside of the reactor, and not by incident light intensity, by combining the results from different studies including ours. This study demonstrated, for the first time, the effectiveness of adding moving bed carriers in photobioreactors to mitigate light inhibition of nitrifiers in a consortium of microalgae and nitrifiers.

High Productivity of Eicosapentaenoic Acid and Fucoxanthin by a Marine Diatom Chaetoceros gracilis in a Semi-Continuous Culture.

Significantly high eicosapentaenoic acid (EPA) and fucoxanthin contents with high production rate were achieved in semi continuous culture of marine diatom. Effects of dilution rate on the production of biomass and high value biocompounds such as EPA and fucoxanthin were evaluated in semi-continuous cultures of Chaetoceros gracilis under high light condition. Cellular dry weight increased at lower dilution rate and higher light intensity conditions, and cell size strongly affected EPA and fucoxanthin contents. The smaller microalgae cells showed significantly higher (p < 0.05) value of 17.1 mg g-dw-1 fucoxanthin and 41.5% EPA content per total fatty acid compared to those observed in the larger cells. Chaetoceros gracilis can accumulate relatively higher EPA and fucoxanthin than those reported previously. In addition, maintenance of small cell size by supplying sufficient nutrients and light energy can be the key for the increase production of valuable biocompounds in C. gracilis.

Microalgae cultivation using undiluted anaerobic digestate by introducing aerobic nitrification-desulfurization treatment.

A novel coupling process using an aerobic bacterial reactor with nitrification and sulfur-oxidization functions followed by a microalgal reactor was proposed for simultaneous biogas desulfurization and anaerobic digestion effluent (ADE) treatment. ADE nitrified by bacteria has a potential to be directly used as a culture medium for microalgae because ammonium nitrogen, including inhibitory free ammonia (NH3), has been converted to harmless NO3 -. To demonstrate this hypothesis, Chlorella sorokiniana NIES-2173, which has ordinary NH3 tolerance; that is, 1.6 mM of EC50 compared with other species, was cultivated using untreated/treated ADE. Compared with the use of a synthetic medium, when using ADE with 1-10-fold dilutions, the specific growth rate and growth yield maximally decreased by 44% and 88%, respectively. In contrast, the algal growth using undiluted ADE treated by nitrification-desulfurization was almost the same as with using synthetic medium. It was also revealed that 50% of PO4 3- and most metal concentrations of ADE decreased following nitrification-desulfurization treatment. Moreover, upon NaOH addition for pH adjustment, the salinity increased to 0.66%. The decrease in metals mitigates the bioconcentration of toxic heavy metals from wastewater in microalgal biomass. Meanwhile, salt stress in microalgae and limiting nutrient supplementation, particularly for continuous cultivation, should be of concern.

Anaerobic digestion effluent treatment using microalgae and nitrifiers in an outdoor raceway pond with fluidized carriers.

Combining microalgae and nitrifiers in a single photobioreactor has attracted attention as an alternative approach for conventional nitrogen removal from wastewater. However, nitrifiers are known to be sensitive to light exposure. This study demonstrated the effectiveness of using fluidized carriers to mitigate light stress in nitrifiers. An outdoor raceway pond containing microalgae and nitrifiers with fluidized carriers was used to treat two-fold diluted anaerobic digestion effluent (785 mg-N L-1 as a form of dissolved total Kjeldahl nitrogen: TKN) over 50 days. The average daily sunlight intensity reached the inhibition level of nitrifiers (423 µmol photons m-2 s-1); however, stable nitrification with a specific ammonium oxidation rate of 55 mg-N g-total suspended solid-1 day-1 was observed. TKN was mostly removed via nitrifier metabolism (ammonium oxidation and uptake: 40.1%) and partially via microalgae uptake (5.7%). Different microalgae-based processes including that of this study were compared in terms of tolerances to a high dissolved TKN concentration and strong light. Our system showed a relatively higher resistance to not only light exposure but also TKN because the nitrification process decreased the free ammonia level to less than 0.25 mg L-1, which allowed microalgae to grow despite the high ammonium concentration.

Effect of Membrane Hydrophobicity and Thickness on Energy-Efficient Dissolved Oxygen Removal From Algal Culture.

Removal of dissolved oxygen from algal photobioreactors is essential for high productivity in mass cultivation. Gas-permeating photobioreactor that uses hydrophobic membranes to permeate dissolved oxygen (pervaporation) from its body itself is an energy-efficient option for oxygen removal. This study comparably evaluated the characteristics of various commercial membranes and determined the criteria for the selection of suitable ones for the gas-permeating photobioreactors. It was found that oxygen permeability is limited not by that in the membrane but in the liquid boundary layer. Membrane thickness had a negative effect on membrane oxygen permeability, but the effect was as minor as less than 3% compared with the liquid boundary layer. Due to this characteristic, the lamination of non-woven fabric with the microporous film did not significantly decrease the overall oxygen transfer coefficient. The permeability in the liquid boundary layer had a significantly positive relationship with the hydrophobicity. The highest overall oxygen transfer coefficients in the water-to-air and water-to-water oxygen removal tests were 2.1 ± 0.03 × 10-5 and 1.39 ± 0.09 × 10-5 m s-1, respectively. These values were considered effective in the dissolved oxygen removal from high-density algal culture to prevent oxygen inhibition. Furthermore, hydrophobicity was found to have a significant relationship also with water entry pressure, which needs to be high to avoid culture liquid leakage. Therefore, these results suggested that a microporous membrane with strong hydrophobicity laminated with non-woven fabric would be suitable characteristics for gas-permeating photobioreactor.

Novel wet-solid states serial anaerobic digestion process for enhancing methane recovery of aquatic plant biomass.

Aquatic plant biomass is characterised by high moisture content and a lignocellulose structure. To apply the anaerobic digestion (AD) treatment to aquatic plants, the simultaneous achievement of high methane (CH4) recovery per biomass volume and high biodegradability have been a challenge owing to these characteristics. Herein, we propose a novel two-stage serial wet- and solid-state AD (SS-AD) system that quickly digests the labile cytoplasm fraction in the first wet AD reactor in a short retention time while slowly digesting the lignocellulosic fraction in the later SS-AD with long retention time. In this study, the effect of this serial AD on CH4 recovery and chemical oxygen demand (COD) balance from aquatic plant biomass was examined in a semi-continuous operation. Elodea nuttallii, which grows excessively in the southern basin of Lake Biwa, Japan, was used as the substrate. For comparison, single-stage AD with different hydraulic retention times (HRTs) (30 d and 15 d) was performed. The CH4 conversion efficiency in single-stage AD deteriorated from 47.6 to 33.1% COD with shortened HRT, probably owing to the low degradability of slowly degrading fraction (i.e. lignocellulose) in the short retention time. In contrast, the serial AD under the same HRT (15 d) as a single-stage AD exhibited higher CH4 conversion efficiency of 65.1% COD, mainly owing to the enhanced degradation of slowly degrading fraction because of the prolonged solid retention time (52.2 d) of the entire system. The CH4 recovery from the wet AD alone in the serial AD system surpassed that from the 30 d-HRT of the single-stage AD, possibly due to the appropriate HRT for labile fraction and/or the microbial recirculation. The serial wet and SS-AD was suggested as a suitable technology for the treatment of aquatic plant biomass with recalcitrant cell walls and a labile cytoplasm.

Incorporation characteristics of exogenous 15N-labeled thymidine, deoxyadenosine, deoxyguanosine and deoxycytidine into bacterial DNA.

Bacterial production has been often estimated from DNA synthesis rates by using tritium-labeled thymidine. Some bacteria species cannot incorporate extracellular thymidine into their DNA, suggesting their biomass production might be overlooked when using the conventional method. In the present study, to evaluate appropriateness of deoxyribonucleosides for evaluating bacterial production of natural bacterial communities from the viewpoint of DNA synthesis, incorporation rates of four deoxyribonucleosides (thymidine, deoxyadenosine, deoxyguanosine and deoxycytidine) labeled by nitrogen stable isotope (15N) into bacterial DNA were examined in both ocean (Sagami Bay) and freshwater (Lake Kasumigaura) ecosystems in July 2015 and January 2016. In most stations in Sagami Bay and Lake Kasumigaura, we found that incorporation rates of deoxyguanosine were the highest among those of the four deoxyribonucleosides, and the incorporation rate of deoxyguanosine was approximately 2.5 times higher than that of thymidine. Whereas, incorporation rates of deoxyadenosine and deoxycytidine were 0.9 and 0.2 times higher than that of thymidine. These results clearly suggest that the numbers of bacterial species which can incorporate exogenous deoxyguanosine into their DNA are relatively greater as compared to the other deoxyribonucleosides, and measurement of bacterial production using deoxyguanosine more likely reflects larger numbers of bacterial species productions.