Bioponic systems with biochar: Insights into nutrient recovery, heavy metal reduction, and microbial interactions in digestate-based bioponics.

Bioponics is a nutrient-recovery technology that transforms nutrient-rich organic waste into plant biomass/bioproducts. Integrating biochar with digestate from anaerobic wastewater treatment process can improve resource recovery while mitigating heavy metal contamination. The overarching goal of this study was to investigate the application of biochar in digestate-based bioponics, focusing on its efficacy in nutrient recovery and heavy metal removal, while also exploring the microbial community dynamics. In this study, biochar was applied at 50 % w/w with 500 g dry weight of digestate during two 28-day crop cycles (uncontrolled pH and pH 5.5) using white stem pak choi (Brassica rapa var. chinensis) as a model crop. The results showed that the digestate provided sufficient phosphorus and nitrogen, supporting plant growth. Biochar amendment improved plant yield and phosphate solubilization and reduced nitrogen loss, especially at the pH 5.5. Furthermore, biochar reduced the heavy metal accumulation in plants, while concentrating these metals in the residual sludge. However, owing to potential non-carcinogenic and carcinogenic health risks, it is still not recommended to directly consume plants cultivated in digestate-based bioponic systems. Additionally, biochar amendment exhibited pronounced impact on the microbial community, promoting microbes responsible for nutrient solubilization and cycling (e.g., Tetrasphaera, Herpetosiphon, Hyphomicrobium, and Pseudorhodoplanes) and heavy metal stabilization (e.g., Leptolinea, Fonticella, Romboutsia, and Desulfurispora) in both the residual sludge and plants. Overall, the addition of biochar enhanced the microbial community and facilitated the metal stabilization and the cycling of nutrients within both residual sludge and root systems, thereby improving the overall efficiency of the bioponics.

Effects of vermicompost leachate on nitrogen, phosphorus, and microbiome in a food waste bioponic system.

Food waste is rich in nutrients, such as nitrogen and phosphorus, and can be integrated with bioponics, a closed-loop agricultural system that combines hydroponics with biological nutrient recovery. Vermicompost leachate (VCL) supplementation has been shown to improve the co-composting of organic waste (i.e., compost quality) and the biodegradation of organic compounds. Thus, VCL has high potential for enhancing nutrient availability in bioponics from food waste. However, the understanding of nitrogen and phosphorus availability in food waste-based bioponics is limited, both with and without VCL. In this study, food waste derived from cafeteria vegetable waste was used as the substrate (500 g dry wt./system) in bioponics to grow lettuce (Lactuca sativa L.) for two consecutive cycles (35 days/cycle) without substrate replacement. VCL was applied weekly (1-5% v/v) and compared to the control without VCL. The results showed that the food waste in bioponics provided nitrogen and phosphorus for plant growth (15.5-65.8 g/lettuce head). Organic-degrading and nutrient-transforming bacteria (Hydrogenispora, Clostridium_sensu_stricto_1, Ruminiclostridium_1, Cellvibrio, Thauera, Hydrogenophaga, and Bacillus) were predominantly found in plant roots and residual food waste. VCL addition significantly increased nitrate, phosphate, and chemical oxygen demand levels in bioponics, owing to the nutrients in VCL and the enhancement of keystone microorganisms responsible for organic degradation and nutrient cycling (e.g., Ellin6067, Actinomyces, and Pirellula). These findings suggest that nitrogen, phosphorus, and organic carbon concentrations in an ecosystem of nutrient-transforming and organic-degrading microbes are key in managing nutrient recovery from food waste in bioponics.

Effects of Cu and Zn contamination on chicken manure-based bioponics: Nitrogen recovery, bioaccumulation, microbial community, and health risk assessment.

In bioponics, although chicken manure is an efficient substrate for vegetable production and nitrogen recovery, it is often contaminated with high Cu and Zn levels, which could potentially cause bioaccumulation in plants and pose health risks. The objectives of this study were to assess nitrogen recovery in lettuce- and pak choi-based bioponics with Cu (50-150 mg/kg) and Zn (200-600 mg/kg) supplementation, as well as their bioaccumulation in plants, root microbial community, and health risk assessment. The supplementation of Cu and Zn did not affect nitrogen concentrations and plant growth (p > 0.05) but reduced nitrogen use efficiency. Pak choi showed higher Cu and Zn bioconcentration factors than lettuce. Bacterial genera Ruminiclostridium and WD2101_soil_group in lettuce roots and Mesorhizobium in pak choi roots from Cu and Zn supplemented conditions were significantly higher (p < 0.05) than controls, suggesting microbial biomarkers in plant roots from Cu and Zn exposure bioponics depended on plant type. Health risk assessment herein revealed that consumption of bioponic vegetables with Cu and Zn contamination does not pose long-term health risks (hazard index <1) to children or adults, according to the US EPA. This study suggested that vegetable produced from chicken manure-based bioponics has low health risk in terms of Cu and Zn bioaccumulation and could be applied in commercial-scale system for nutrient recovery from organic waste to vegetable production; however, health risk from other heavy metals and xenobiotic compounds must be addressed.

Inhibition of anaerobic ammonium oxidation (anammox) bacteria by addition of high and low concentrations of chloramphenicol and comparison of attached- and suspended-growth.

Inhibition of anammox activities was tested with two ranges of chloramphenicol (CAP) concentration (5, 10, 20, 50, and 100 mg L-1) and (100, 500, and 1000 µg L-1). In a short-term study, strong inhibition of activity was dependent of CAP concentration in both attached-growth (SBR-A) and suspended-growth (SBR-S) systems. The activities of attached-growth cultures at all CAP concentrations were reversible after 1 day, while activities for suspended-growth cultures were only gradually reversible dependent on the CAP concentrations. In long-term studies with daily additions of 6 mg L-1 CAP, the anammox activity on day 41 in SBR-A had decreased to 18% baseline (SAA reduced from 0.528 to 0.096 mg N mg-1 VSS d-1). More rapid reduction of anammox activity was observed in SBR-S, down to 17% baseline after only 27 days (SAA decreased from 0.576 to 0.096 mg N mg-1 VSS d-1). Inhibition was irreversible in both SBR-S and SBR-A after the long-term study. With lower CAP additions (100-1000 µg L-1), the activities in both reactors were stable during daily CAP addition for two weeks. Attached-growth cultures tended to be more tolerant of CAP addition than suspended-growth cultures. Both un-competitive and non-competitive models could be used to compare anammox activities with the higher CAP concentrations. The SAAmax [fx] (the maximum specific anammox activity) and hKi (the inhibition constant) of SBR-A were 0.48 mg N mg-1 VSS d-1 and 98.3 mg L-1, respectively. The SAAmax[fx] and Ki of SBR-S were 1.25 mg N mg-1 VSS d-1and 71.1 mg L-1, respectively.

Resuscitation of starved suspended- and attached-growth anaerobic ammonium oxidizing bacteria with and without acetate.

Anammox application for nutrient removal from wastewater is increasing, though questions remain about anammox resilience to fluctuating conditions. Resuscitation of anammox suspended- and attached-growth cultures after 3 months of starvation was studied with and without acetate dosing. Without acetate, the attached-growth culture recovered more quickly than the suspended-growth culture. Suspended-growth cultures recovered more quickly (within 60 days) with weekly and daily acetate dosing than without, but anammox activity and copy numbers decreased with continued acetate addition. All attached-growth cultures recovered within 60 days, but after that activity with acetate dosing was consistently at least 20% lower than that without acetate addition. Ca. Jettenia caeni, Ca. Anammoxoglobus sp., Ca. Brocadia fulgida, Ca. Brocadia anammoxidans, Ca. Brocadia fulgida and Ca. Jettenia asiatica were identified. Acetate addition can significantly accelerate short-term resuscitation of enriched anammox suspended-growth cultures after starvation but may reduce anammox activity over the longer term in suspended- and attached-growth cultures.

ANAMMOX-like performances for nitrogen removal from ammonium-sulfate-rich wastewater in an anaerobic sequencing batch reactor.

Ammonium removal by the ANaerobic AMonium OXidation (ANAMMOX) process was observed through the Sulfate-Reducing Ammonium Oxidation (SRAO) process. The same concentration of ammonium (100 mg N L(-1)) was applied to two anaerobic sequencing batch reactors (AnSBRs) that were inoculated with the same activated sludge from the Vermicelli wastewater treatment process, while nitrite was fed in ANAMMOX and sulfate in SRAO reactors. In SRAO-AnSBR, in substrates that were fed with a ratio of NH4(+)/SO4(2-) at 1:0.4 ± 0.03, a hydraulic retention time (HRT) of 48 h and without sludge draining, the Ammonium Removal Rate (ARR) was 0.02 ± 0.01 kg N m(-3).d(-1). Adding specific ANAMMOX substrates to SRAO-AnSBR sludge in batch tests results in specific ammonium and nitrite removal rates of 0.198 and 0.139 g N g(-1) VSS.d, respectively, indicating that the ANAMMOX activity contributes to the removal of ammonium in the SRAO process using the nitrite that is produced from SRAO. Nevertheless, the inability of ANAMMOX to utilize sulfate to oxidize ammonium was also investigated in batch tests by augmenting enriched ANAMMOX culture in SRAO-AnSBR sludge and without nitrite supply. The time course of sulfate in a 24-hour cycle of SRAO-AnSBR showed an increase in sulfate after 6 h. For enriched SRAO culture, the uptake molar ratio of NH4(+)/SO4(2-) at 8 hours in a batch test was 1:0.82 lower than the value of 1:0.20 ± 0.09 as obtained in an SRAO-AnSBR effluent, while the stoichiometric ratio of 1:0.5 that includes the ANAMMOX reaction was in this range. After a longer operation of more than 2 years without sludge draining, the accumulation of sulfate and the reduction of ammonium removal were observed, probably due to the gradual increase in the sulfur denitrification rate and the competitive use of nitrite with ANAMMOX. The 16S rRNA gene PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and PCR cloning analyses resulted in the detection of the ANAMMOX bacterium (Candidatus Brocadia sinica JPN1) Desulfacinum subterraneum belonging to the genus Desulfacinum and bacteria that are involved in sulfur metabolism (Pseudomonas aeruginosa strain SBTPe-001 and Paracoccus denitrificans strain IAM12479) in SRAO-AnSBR.

Identification and quantification of bacteria and archaea responsible for ammonia oxidation in different activated sludge of full-scale wastewater treatment plants.

In this study, the abundance and sequences of the amoA gene in ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA) were defined in three wastewater treatment plants using activated sludge with biological nitrogen removal in different countries: Thailand, United States of America (USA), and Japan. Quantitative real-time polymerase chain reaction (PCR) and PCR coupled with denaturing gradient gel electrophoresis were used to find the comparative abundance and identity of AOB and AOA. The conditions at the Phuket WWTP in Thailand promoted the dominance of AOA amoA genes over AOB amoA genes, while conditions at the WWTPs in Japan and USA promoted growth of AOB. Three parameters that may have contributed to the AOA dominance in Phuket were longer SRT, higher temperature, and higher pH. The Phuket WWTP is a unique system that can be used to better understand the conditions that promote AOA growth and dominance over AOB. In addition, analysis of operational data in conjunction with AOA and AOB community structure from the Phuket WWTP may elucidate advantages of AOA in meeting stricter treatment standards.

Comparison of nitrogen removal rates and nitrous oxide production from enriched anaerobic ammonium oxidizing bacteria in suspended and attached growth reactors.

Attached growth-systems for the anaerobic ammonium oxidation (anammox) process have been postulated for implementation in the field. However, information about the anammox process in attached growth-systems is limited. This study compared nitrogen removal rates and nitrous oxide (N2O) production of enriched anammox cultures in both suspended and attached growth sequencing batch reactors (SBRs). Suspended growth reactors (SBR-S) and attached growth reactors using polystyrene sponge as a medium (SBR-A) were used in these experiments. After inoculation with an enriched anammox culture, significant nitrogen removals of ammonium (NH4 (+)) and nitrite (NO2 (-)) were observed under NH4 (+):NO2 (-) ratios ranging from 1:1 to 1:2 in both types of SBRs. The specific rates of total nitrogen removal in SBR-S and SBR-A were 0.52 mg N/mg VSS-d and 0.44 mg N/mg VSS-d, respectively, at an NH4 (+):NO2 (-) ratio of 1:2. N2O production by the enriched anammox culture in both SBR-S and SBR-A was significantly higher at NH4 (+):NO2 (-) ratio of 1:2 than at NH4 (+):NO2 (-) ratios of 1:1 and 1:1.32. In addition, N2O production was higher at a pH of 6.8 than at pH 7.3, 7.8, and 8.3 in both SBR-S and SBR-A. The results of this investigation demonstrate that the anammox process may avoid N2O emission by maintaining an NH4 (+):NO2 (-) ratio of less than 1:2 and pH higher than 6.8.

Effects of oxytetracycline on anammox activity.

Batch experiments were conducted to investigate the effects of oxytetracycline on anaerobic ammonium oxidation (anammox) process. The short-and long-term effects on anammox activity were studied by measuring ammonium (NH(4)(+)), nitrite (NO(2)(-)), and nitrate (NO(3)(-)) concentrations over time. Experiments were conducted at NH(4)(+)and NO(2)(-) concentrations of 60-90 mg N/L and 60-190 mg N/L, respectively (NH(4)(+):NO(2)(-) ratio from 1:1-1:2.25), oxytetracycline concentrations of 10-100 mg/L, and biomass concentrations of 300-800 mg/L. In the short-term study, anammox activity was inhibited by all oxytetracycline concentrations studied. However, daily addition of oxytetracycline to a concentration of 5 ± 3.5 mg/L in the anammox sequencing batch reactor completely inhibited anammox activity in the fifth week. Fluorescent in situ hybridization was used to identify autotrophic ammonium oxidizing bacteria (Nitrosomonas spp., Nitrobacter spp., Nitrospira spp., Candidatus Brocadia anammoxidans, and Candidatus Kuenenia stuttgartiensis). The population of anammox culture was significantly decreased while Nitrosomonas spp. and Nitrospira spp. increased in the fifth week compared with the first week of experiment. A not-competitive model fit the anammox inhibition data at oxytetracycline concentrations of 0-100 mg/L quite well with V(max) of 0.0435 mg N/mg VSS-hr and K(i) of 54.66 mg/L.

Nitrogen removal of anammox cultures under different enrichment conditions.

Anammox bacteria in sludge from an anoxic tank of a municipal wastewater treatment plant at Nongkhaem, Bangkok, Thailand were enriched in two sequencing batch reactors (SBRs; SBR-1 and SBR-2), under different conditions. SBR-1 was open to the atmosphere, while SBR-2 was closed and flushed with a mixture of 95% argon and 5% CO(2) during the fill period in order to provide strict anaerobic conditions. The specific nitrogen removal rates of SBR-1 and SBR-2 were 0.43 g N/g VSS-d and 2.59 g N/g VSS-d, respectively. Denaturing gradient gel electrophoresis (DGGE) analyses showed differences in band patterns among the Nongkhaem sludge and the two enrichment cultures. Based on fluorescent in situ hybridization (FISH), the anammox bacteria in both systems were either "Candidatus Brocadia anammoxidans" or "Candidatus Kuenenia stuttgartiensis". The results from this study demonstrate the potential of alternative anammox systems for nitrogen removal and provide information on the microbial communities of anammox cultures under different enrichment conditions.