intI1 gene abundance from septic tanks in Thailand using validated intI1 primers.

IMPORTANCE: Antimicrobial resistance is a global crisis, and wastewater treatment, including septic tanks, remains an important source of antimicrobial resistance (AMR) genes. The role of septic tanks in disseminating class 1 integron, and by extension AMR genes, in Thailand, where antibiotic use is unregulated remains understudied. We aimed to monitor gene abundance as a proxy to infer potential AMR from septic tanks in Thailand. We evaluated published intI1 primers due to the lack of consensus on optimal Q-PCR primers and the absence of standardization. Our findings confirmed septic tanks are a source of class 1 integron to the environment. We highlighted the significance of intI1 primer choice, in the context of interpretation of risk associated with AMR spread from septic tanks. We recommend the validated set (F3-R3) for optimal intI1 quantification toward the goal of achieving standardization across studies.

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.

Comparative study on attached-growth photobioreactors under blue and red lights for treatment of septic tank effluent.

Attached-growth photobioreactors (AG-PBRs) employing low-cost attached-growth media were applied to treat septic tank effluent which contained abundant organic and nutrient matters as well as pathogenic microorganisms. This study investigated effects of blue and red LED lights on organic, nutrient and pathogenic removals, biomass productivity and compositions of microbial community in the AG-PBR system. The experimental results showed the blue AG-PBR to be more effective in removing chemical oxygen demand (COD), total nitrogen (TN) and ammonia nitrogen (NH3-N) and generating biomass productivity than those of the red AG-PBR (P < 0.05). Mass balance analysis indicated that the TN and total phosphorus (TP) were removed mainly by assimilation into the biomass. The TN removal rates via nitrification and denitrification processes in the blue AG-PBR were found to be higher than that of the red AG-PBR, corresponding to the observed results of bacterial biomass and abundances of nitrifying and denitrifying bacterial species in the treatment systems. The maximal areal algal biomass productivity of 47 gDW/(m2. d) in the blue AG-PBRs was found to be higher than those of other algal attached-growth systems. Although, the red and blue AG-PBR systems could effectively treat the septic tank effluent to meet the national and international discharge standards, based on treatment efficiencies and biomass productivity, the blue AG-PBR is recommended for treatment of septic tank effluent.

Performance evaluation of novel attached-growth high rate algal pond system with additional artificial illumination for wastewater treatment and nutrient recovery.

Domestic wastewater containing a high proportion of organic matter and nutrients is a serious pollution problem in developing countries. This study aimed to evaluate the performance of a novel attached-growth high rate algal pond (AG-HRAP) employing attached-growth media and artificial light sources for treating domestic wastewater and enhancing nutrient recovery. Light intensities in the range of 40-180 µmol/m2/s were used in the AG-HRAPs. The experimental results showed that the highest chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) removal efficiencies of 88, 62 and 69%, respectively, were found at the hydraulic retention time (HRT) of 15 days and the average light intensity of 180 µmol/m2/s. Moreover, the effluent COD concentrations could meet Thailand's national discharge standard. The highest biomass and protein productivities of 54 ± 4 and 37 ± 8 g/m2/d, respectively, were found in the AG-HRAPs, which were higher than in previous studies of HRAPs. The Stover-Kincannon kinetic values for COD, TN and TP removals of the AG-HRAPs (R2 = 0.9) were higher than those of the conventional systems. Additionally, the novel AG-HRAP system could provide a highly cost-effective operation when compared to other microalgal systems.

Removal and monitoring acetaminophen-contaminated hospital wastewater by vertical flow constructed wetland and peroxidase enzymes.

Hospital wastewater contains acetaminophen (ACT) and nutrient, which need adequate removal and monitoring to prevent impact to environment and community. This study developed a pilot scale vertical flow constructed wetland (CW) to (1) remove high-dose ACT and pollutants in hospital wastewater and (2) identify the correlation of peroxidase enzyme extruded by Scirpus validus and pollutants removal efficiency. By that correlation, a low-cost method to monitor pollutants removal was drawn. Plants, such as Scirpus validus, generated peroxidase enzymes to alleviate pollutants' stress. Results showed that the CW removed 3.5 to 6 logs of initial concentration 10 mg ACT/L to a recommended level for drinking water. The CW eliminated COD, TKN and TP efficiently, meeting the wastewater discharged standards of Thailand and Vietnam. By various multivariable regression models, concentrations of ACT in CW effluent and enzymes in S. validus exhibited a significant correlation (p < 0.01, R2 = 68.3%). These findings suggested that (i) vertical flow CW could remove high-dose ACT and nutrient and (ii) peroxidase enzymes generated in S. validus, such as soluble and covalent ones, could track ACT removal efficiency. This would help to reduce facilities and analytical cost of micro-pollutants.

Faecal sludge treatment and utilization by hydrothermal carbonization.

Hydrothermal carbonization (HTC) is a thermal conversion process that can be applied to convert faecal sludge into carbonaceous solids, called hydrochar. In this study, the technical feasibility of hydrochar production by HTC of faecal sludge was investigated. Experimental results showed energy contents of the produced hydrochar to be about 19-20 MJ/kg, comparable to natural coals and therefore usable as a solid fuel. The produced hydrochar contained a carbon content of approximately 40%wt, which could be processed further to make it suitable as an anode in batteries. The produced hydrochar also had adsorption characteristics for removing heavy metals and micropollutants in wastewater. Liquid by-products obtained from the HTC process were found to contain high concentrations of organic matter, while the amount of gas produced was 10 L-gas/kg-FS with CO2 is the main component. The bio-methane potential tests of this liquid product suggested the methane production of about 2.0 L-CH4 per kg-faecal sludge could be obtained.

Performance and kinetics of algal-bacterial photobioreactor (AB-PBR) treating septic tank effluent.

Septic tank effluent contains high organic and nutrient contents. This study aimed to evaluate treatment performance of an algal-bacterial photobioreactor (AB-PBR) treating the septic tank effluent. The experimental unit employed a transparent plastic medium made from recycled drinking water bottles for attached-growth biofilm. Red LED lamp (light intensity ∼100 µmol/m2/s) was applied as an energy source for the growth of algal-bacterial biofilm in the AB-PBR. The experimental results showed that AB-PBR operated at the hydraulic retention time (HRT) of 3 days gave the highest chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) removal efficiencies of 64, 45 and 35%, respectively, by which the effluent COD concentrations could meet the effluent standards of Thailand, but the effluent TN and TP concentrations needed to be further removed. The Stover-Kincannon model was applied to determine the kinetic values of COD and TN removals with R2 values greater than 0.8. Microbiological examinations indicated Chlorella sp. is the predominant algal species growing in the AB-PBR, while the amplicon sequencing information analytical results revealed the bacterial phylum of Proteobacteria to be the predominant bacterial group.

Rapid and Decentralized Human Waste Treatment by Microwave Radiation.

This study evaluates the technical feasibility of using microwave radiation for the rapid treatment of human feces. Human feces of 1000 g were radiated with a commercially available household microwave oven (with rotation) at different exposure time lengths (30, 50, 60, 70, and 75 mins) and powers (600, 800, and 1000 W). Volume reduction over 90% occurred after 1000 W microwave radiation for 75 mins. Pathogen eradiation performances of six log units or more at a high range of microwave powers were achieved. Treatments with the same energy input of 1000 Wh, but at lower powers with prolonged exposure times, significantly enhanced moisture removal and volume reduction. Microwave radiation caused carbonization and resulted in a more stable end product. The energy content of the samples after microwave treatment at 1000 W and 75 mins is 3517 ± 8.85 calories/g of dried sample, and the product can also be used as compost.

Enhanced sludge reduction in septic tanks by increasing temperature.

Septic tanks in most developing countries are constructed without drainage trenches or leaching fields to treat toilet wastewater and /or grey water. Due to the short hydraulic retention time, effluents of these septic tanks are still highly polluted, and there is usually high accumulation of septic tank sludge or septage containing high levels of organics and pathogens that requires frequent desludging and subsequent treatment. This study aimed to reduce sludge accumulation in septic tanks by increasing temperatures of the septic tank content. An experimental study employing two laboratory-scale septic tanks fed with diluted septage and operating at temperatures of 40 and 30°C was conducted. At steady-state conditions, there were more methanogenic activities occurring in the sludge layer of the septic tank operating at the temperature of 40°C, resulting in less total volatile solids (TVS) or sludge accumulation and more methane (CH4) production than in the unit operating at 30°C. Molecular analysis found more abundance and diversity of methanogenic microorganisms in the septic tank sludge operating at 40°C than at 30°C. The reduced TVS accumulation in the 40°C septic tank would lengthen the period of septage removal, resulting in a cost-saving in desluging and septage treatment. Cost-benefit analysis of increasing temperatures in septic tanks was discussed.

A model for methane production in sewers.

Most sewers in developing countries are combined sewers which receive stormwater and effluent from septic tanks or cesspools of households and buildings. Although the wastewater strength in these sewers is usually lower than those in developed countries, due to improper construction and maintenance, the hydraulic retention time (HRT) could be relatively long and resulting considerable greenhouse gas (GHG) production. This study proposed an empirical model to predict the quantity of methane production in gravity-flow sewers based on relevant parameters such as surface area to volume ratio (A/V) of sewer, hydraulic retention time (HRT) and wastewater temperature. The model was developed from field survey data of gravity-flow sewers located in a peri-urban area, central Thailand and validated with field data of a sewer system of the Gold Coast area, Queensland, Australia. Application of this model to improve construction and maintenance of gravity-flow sewers to minimize GHG production and reduce global warming is presented.

Performance evaluation of modified Living Wall garden for treating septic tank effluent.

The Living Wall (LW) garden system has been employed as a post-treatment system to improve the effluent quality of septic tanks. This improvement primarily involves reducing nutrient levels, as well as facilitating the removal of organic matter and solids in accordance with effluent discharge guidelines. The objective of this study was to investigate the treatment performance of the LW system connected to a septic tank, along with an examination of the microbial communities within the LW units. A laboratory-scale LW system, comprising LW1, LW2, and LW3 units, was employed. The system was fed with effluent obtained from septic tanks and varied by theoretical hydraulic retention time (HRT) of 6, 12, and 24 h. The TCOD, SCOD, TSS, TVS, TKN, and TP removal efficiencies of the LWs were achieved at 62 ± 24, 42 ± 19, 72 ± 21, 66 ± 15, 80 ± 15, and 58 ± 21%, respectively. To classify microbial communities in the soil and gravels collected from each LW unit, the Illumina MiSeq System Sequencer was employed. Nitrospirota was consistently found in all LW units, aiding in the conversion of nitrogen. Fusobacteriota were detected in specific layers of the LW units, indicating varying oxygen levels in the LW system.

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.

A concept for planning and management of on-site and centralised municipal wastewater treatment systems, a case study in Bangkok, Thailand. I: pollutant discharge indicators and pollutant removal efficiency functions.

The concept of pollution load indicators for planning and management of the mixture conditions of centralised and on-site wastewater treatment systems has not been discussed in detail so far. In this paper, pollutant discharge (load) indicators and pollutant removal efficiencies were quantitatively analysed to develop a part of a strategy for planning and management of municipal wastewater treatment systems (WWTSs) under the mixture conditions in Bangkok, Thailand, as a case study. Pollutant discharge indicators of on-site WWTSs were estimated based on the relevant literature. Three kinds of pollutant removal efficiency function at centralised wastewater treatment plants (CWWTPs) were empirically developed for biological oxygen demand, chemical oxygen demand, total nitrogen, total phosphorus, total coliforms and faecal coliforms based on the existing CWWTP management data. These results will be integrated into the scenario-based analysis in the second paper in the series. The results will be base datasets, and the concept and estimation methods can be applied for wastewater treatment planning and management in other areas.

A concept for planning and management of on-site and centralised municipal wastewater treatment systems, a case study in Bangkok, Thailand. II: scenario-based pollutant load analysis.

Scenario-based pollutant load analysis was conducted to develop a part of a concept for planning and management of wastewater treatment systems (WWTSs) under the mixture conditions of centralised and on-site WWTSs. Pollutant discharge indicators and pollutant removal efficiency functions were applied from another paper in the series, which were developed based on the existing conditions in urban and peri-urban areas of Bangkok, Thailand. Two scenarios were developed to describe development directions of the mixture conditions. Scenario 1 involves keeping the on-site wastewater treatment plants (WWTPs) within the areas of centralised WWTSs. Scenario 2 is dividing the centralised and on-site WWTS areas. Comparison of the smallest values of total pollutant discharge per capita (PDCtotal) between Scenarios 1 and 2 showed that the smallest PDCtotal in Scenario 1 was smaller than that in Scenario 2 for biological oxygen demand, chemical oxygen demand and total phosphorus whereas the smallest PDCtotal in Scenario 2 was smaller than that in Scenario 1 for total nitrogen, total coliforms and faecal coliforms. The results suggest that the mixture conditions could be a possible reason for smaller pollutant concentrations at centralised WWTPs. Quantitative scenario-based estimation of PDCtotal is useful and a prerequisite in planning and management of WWTSs.

Comparison of rapid methods for detection of Giardia spp. and Cryptosporidium spp. (oo)cysts using transportable instrumentation in a field deployment.

Reliable, sensitive, quantitative, and mobile rapid screening methods for pathogenic organisms are not yet readily available, but would provide a great benefit to humanitarian intervention units in disaster situations. We compared three different methods (immunofluorescent microscopy, IFM; flow cytometry, FCM; polymerase chain reaction, PCR) for the rapid and quantitative detection of Giardia lamblia and Cryptosporidium parvum (oo)cysts in a field campaign. For this we deployed our mobile instrumentation and sampled canal water and vegetables during a 2 week field study in Thailand. For purification and concentrations of (oo)cysts, we used filtration and immunomagnetic separation. We were able to detect considerably high oo(cysts) concentrations (ranges: 15-855 and 0-240 oo(cysts)/liter for Giardia and Cryptosporidium, respectively) in 85 to 300 min, with FCM being fastest, followed by PCR, and IFM being slowest due to the long analysis time per sample. FCM and IFM performed consistently well, whereas PCR reactions often failed. The recovery, established by FCM, was around 30% for Giardia and 13% for Cryptosporidium (oo)cysts. It was possible to track (oo)cysts from the wastewater further downstream to irrigation waters and confirm contamination of salads and water vegetables. We believe that rapid detection, in particular FCM-based methods, can substantially help in disaster management and outbreak prevention.

Fate identification and management strategies of non-recyclable plastic waste through the integration of material flow analysis and leakage hotspot modeling.

Low priority on waste management has impacted the complex environmental issue of plastic waste pollution, as evident by results of this study where it was found that 24.3% of waste generation in Jakarta and Bandung is emitted into the waterway due to the high intensity of human activity in the urban area. In this study, we investigated the viable integration between material flow analysis and leakage hotspot modeling to improve management strategies for plastic pollution in water systems and open environments. Using a multi-criteria assessment of plastic leakage from current waste management, a material flow analysis was developed on a city-wide scale defining the fate of plastic waste. Geospatial analysis was assigned to develop a calculation for identification and hydrological analysis while identifying the potential amount of plastic leakage to the river system. The results show that 2603 tons of plastic accumulated along the mainstream of the Ciliwung River on an annual basis, and a high-density population like that in Bandung discarded 1547 tons in a one-year period to the Cikapundung River. The methods and results of this study are applicable towards improving the control mechanisms of river rejuvenation from plastic leakage by addressing proper management in concentrated locations.

Chicken manure-based bioponics: Effects of acetic acid supplementation on nitrogen and phosphorus recoveries and microbial communities.

Bioponics has the potential to recover nutrients from organic waste streams, such as chicken manure and digestate with high volatile fatty acid (VFA) contents through crop production. Acetic acid, a dominant VFA, was supplemented weekly (0, 500, 1000, and 1500 mg/L) in a chicken manure-based bioponic system, and its effect on the performance of bioponics (e.g., plant yield and nitrogen and phosphorus availabilities) was examined. Microbial communities were analyzed using 16S rRNA gene sequencing, and the functional gene abundances were predicted using PICRUSt. Although acetic acid negatively affected plant yield, no significant difference (p > 0.05) was noted in the average nitrogen or phosphorus concentration. In terms of nutrient recovery, the bioponic systems still functioned well, although higher concentrations of acetic acid decreased plant yield and altered the bacterial communities in plant roots and chicken manure sediments. These data suggest that an acetic acid concentration of < 500 mg/L or a longer loading interval is recommended for the effective operation of chicken manure and digestate-based bioponics.

Performance of novel constructed wetlands for treating solar septic tank effluent.

To improve treatment performance of the solar septic tank technology, novel constructed wetland systems have been proposed as an effective post-treatment system. This study aimed to investigate the treatment performance of the multi-soil layer based constructed wetland (MSL-CW) and comparing with the modified constructed wetland (mCW) for treating solar septic tank effluent in long-term operation. Pilot-scale MSL-CW and mCW units were operated in parallel under the same conditions during the period of 2016-2019. Removal efficiencies of TCOD, SCOD and TBOD in the MSL-CW were not significantly different (p < 0.05) from those of the mCW unit, which were 70-72%, 63-68% and 78-82%, respectively. The removal efficiencies of TSS, TKN, NH4-N and TP were found in the same magnitude in both units. The total coliform and E.coli counts in the effluent of MSL-CW and mCW units were reduced from 105 MPN/100 mL to be lower than 103 MPN/100 mL. These long-term operational results demonstrated that the effluent from the MSL-CW and mCW units could meet the global standards of non-sewered sanitation systems and the WHO guidelines. The effects of seasonal variations and plant harvesting on the monthly treatment performance are discussed in this study.

Bioponic system for nitrogen and phosphorus recovery from chicken manure: Evaluation of manure loading and microbial communities.

Bioponics integrates the biological treatment of nutrient-rich waste streams with hydroponics. However, there are several challenges of bioponics, especially nutrient availability and qualities, which affect plant yield. In this study, chicken manure based-nutrient film technique bioponics was examined at manure loadings of 200, 300, and 400 g dry wt. per bioponic system (total of 18 plants). Bioponics effectively released nitrogen and phosphorus (total ammonia nitrogen of 5.8-8.0 mgN/L, nitrate of 7.0-11.2 mgN/L, and phosphate of 48.7-74.2 mgP/L) for efficient growth of lettuce (Lactuca sativa; total yield of 1208-2030 g wet wt. per 18 plants). Nitrogen and phosphorus use efficiencies were 35.1-41.8% and 6.8-8.0%, respectively, and were comparable to aquaponics. Next-generation sequencing was used to examine the microbial communities in digested chicken manure and plant roots in bioponics. Results showed that several microbial genera were associated with organic degradation (e.g., Nocardiopsis spp., Cellvibrio spp.), nitrification (Nitrospira spp.), phosphorus solubilization, and plant growth promotion (e.g., WD2101_soil_group, and Bacillus spp.). Nocardiopsis spp., Romboutsia spp. and Saccharomonospora spp. were found at high abundances and a high degree of co-occurrences among the microbiota, suggesting that the microbial organic decomposition to nitrogen and phosphorus release could be the key factors to achieve better nutrient recovery in bioponics.

Hydraulic characteristics of an anaerobic baffled reactor as onsite wastewater treatment system.

The feasibility of using anaerobic baffled reactor (ABR) as onsite wastewater treatment system was discussed. The ABR consisted of one sedimentation chamber and three up-flow chambers in series was experimented under different peak flow factors (PFF of 1 to 6), superficial gas velocities (between 0.6 and 3.1 cm/hr) and hydraulic retention times (HRT) (24, 36 and 48 hr). Residence time distribution (RTD) analyses were carried out to investigate the hydraulic characteristics of the ABR. It was found that the PFF resulted in hydraulic dead space. The dead space did not exceed 13% at PFF of 1, 2 and 4 while there was 2-fold increase (26%) at PFF of 6. Superficial gas velocities did not result in more (biological) dead space. The mixing pattern of ABR tended to be a completely-mixed reactor when PFF increased. Superficial gas velocities did not affect mixing pattern. The effects of PFF on mixing pattern could be minimized by higher HRT (48 hr). The tank-in-series (TIS) model (N = 4) was suitable to describe the hydraulic behaviour of the studied system. The HRT of 48 hr was able to maintain the mixing pattern under different flow patterns, introducing satisfactory hydraulic efficiency. Chemical oxygen demand (COD) and total suspended solids (TSS) removals under all flow patterns were achieved more than 85% and 90%, respectively. The standard deviation of effluent COD and TSS concentration did not exceed 15 mg/L.