Duckweed-based Wastewater Treatment Research Articles

Potential of Predatory Bacteria to Colonize the Duckweed Microbiome and Change Its Structure: A Model Study Using the Obligate Predatory Bacterium, Bacteriovorax sp. HI3.

Modifying the duckweed microbiome is a major challenge for enhancing the effectiveness of duckweed-based wastewater treatment and biomass production technologies. We herein examined the potential of the exogenous introduction of predatory bacteria to change the duckweed microbiome. Bacteriovorax sp. HI3, a model predatory bacterium, colonized the core of the Lemna microbiome, and its predatory behavior changed the microbiome structure, which correlated with colonization density. These results reveal that bacterial predatory interactions may be important drivers that shape the duckweed microbiome, suggesting their potential usefulness in modifying the microbiome.

Creating a Circular Nitrogen Bioeconomy in Agricultural Systems through Nutrient Recovery and Upcycling by Microalgae and Duckweed: Past Efforts and Future Trends

HighlightsAquatic vegetation-based nutrient recovery offers an alternate approach for treating agricultural wastewater.Microalgae and duckweed can upcycle waste nutrients into valuable bio-based products.Producing feed, fertilizer, and fuel from manure-grown aquatic vegetation promotes a circular N-bioeconomy.Abstract. The massive amounts of nutrients that are currently released into the environment as waste have the potential to be recovered and transformed from a liability into an asset through photosynthesis, industry insight, and ecologically informed engineering design aimed at circularity. Fast-growing aquatic plant-like vegetation such as microalgae and duckweed have the capacity to enable local communities to simultaneously treat their own polluted water and retain nutrients that underlie the productivity of modern agriculture. Not only are they highly effective at upcycling waste nutrients into protein-rich biomass, microalgae and duckweed also offer excellent opportunities to substitute or complement conventional synthetic fertilizers, feedstocks in biorefineries, and livestock feed while simultaneously reducing the energy consumption and greenhouse gas emissions that would otherwise be required for their production and transport to farms. Integrated systems growing microalgae or duckweed on manure or agricultural runoff, and subsequent reuse of the harvested biomass to produce animal feed, soil amendments, and biofuels, present a sustainable approach to advancing circularity in agricultural systems. This article provides a review of past efforts toward advancing the circular nitrogen bioeconomy using microalgae- and duckweed-based technologies to treat, recover, and upcycle nutrients from agricultural waste. The majority of the work with microalgae- and duckweed-based wastewater treatment has been concentrated on municipal and industrial effluents, with <50% of studies focusing on agricultural wastewater. In terms of scale, more than 91% of the microalgae-based studies and 58% of the duckweed-based studies were conducted at laboratory-scale. While the range of nutrient removals achieved using these technologies depends on various factors such as species, light, and media concentrations, 65% to 100% of total N, 82% to 100% of total P, 98% to 100% of NO3-, and 96% to 100% of NH3/NH4+ can be removed by treating wastewater with microalgae. For duckweed, removals of 75% to 98% total N, 81% to 93% total P, 72% to 98% NH3/NH4+, and 57% to 92% NO3- have been reported. Operating conditions such as hydraulic retention time, pH, temperature, and the presence of toxic nutrient levels and competing species in the media should be given due consideration when designing these systems to yield optimum benefits. In addition to in-depth studies and scientific advancements, policies encouraging supply chain development, market penetration, and consumer acceptance of these technologies are vitally needed to overcome challenges and to yield substantial socio-economic and environmental benefits from microalgae- and duckweed-based agricultural wastewater treatment. Keywords: Circular bioeconomy, Duckweed, Manure treatment, Microalgae, Nitrogen, Nutrient recycling, Wastewater treatment.

Simultaneous boron (B) removal and electricity generation from domestic wastewater using duckweed-based wastewater treatment reactors coupled with microbial fuel cell

Boron removal from water environment is a critical issue for scientific spotlight because its removal from wastewater is difficult and costly with conventional treatment method. Herein, an innovative, cost effective and attractive method which depends on duckweed-based wastewater treatment systems coupled with microbial fuel cell reactor (DWWT-MFC) was investigated for B-polluted domestic wastewater treatment and simultaneous electricity generation for the first time in an eco-technological study. Lemna gibba L. was selected as a model duckweed species, and different reactors were also designed to identify which mechanisms are dominant for B removal in a DWWT-MFC reactor matrix. DWWT-MFC reactor achieved 71% B removal in experiment period, and the plant effect on B removal mechanisms in the reactor matrix was recorded as 37.7 ± 4.92% (F = 2.543, p < 0.05). However, supplementary aeration and microbial effects on B removal were determined as negligible. Average maximum voltage output was found as 1.47 V, and maximum power density was 34.8 mW/m2 at a current density of 43.9 mA/m2 with supplementary aeration. Moreover, DWWT-MFC reactor achieved 84%, 81% and 76% of COD, NH4+ and PO43− removal efficiencies, respectively. Moreover, L. gibba grew well in the anode chamber of DWWT-MFC with an average biomass yield of 218 ± 43 g/m2 and a total chlorophyll (a+b) concentration of 30.2 mg g−1, which indicates that anolyte environment was not toxic for L. gibba growth. Consequently, it can be suggested that environmental experts may use DWWT-MFC as an efficient removal method to treat B from domestic wastewater and to produce bioelectricity.

Investigation of biomass production, crude protein and starch content in laboratory wastewater treatment systems planted with Lemna minor and Lemna gibba

ABSTRACTThe use of duckweed-based wastewater treatment systems for producing biomass with high crude protein and starch content was investigated in the current study. For this reason, three lab-scale systems were used; System 1 was planted with Lemna minor, System 2 with Lemna gibba and System 3 with the combination of the two duckweeds. The studied duckweeds were cultivated using secondary treated wastewater as substrate (Phase A), in the presence of excess NH4-N (Phase B) and using water with no nutrients (Phase C). All systems achieved average NH4-N removal higher that 90%. The specific duckweeds growth rates and the specific duckweeds growth rates normalized to the area ranged between 0.14 d−1 and 8.9 g m−2 d−1 (System 1) to 0.19 d−1 and 14.9 g m−2 d−1(System 3). The addition of NH4-N resulted in a significant increase of biomass protein content, reaching 44.4% in System 3, 41.9% in System 2 and 39.4% in System 1. The transfer of biomass in water containing no nutrients resulted in the gradual increment of the starch content up to the end of the experiment. The highest starch content was achieved for the combination of the two duckweeds (46.1%), followed by L. gibba (44.9%) and L. minor (43.9%).

Enhanced biomass production of duckweeds by inoculating a plant growth-promoting bacterium, Acinetobacter calcoaceticus P23, in sterile medium and non-sterile environmental waters.

Duckweed offers the promise of a co-benefit culture combining water purification with biomass production. Acinetobacter calcoaceticus P23 is a plant growth-promoting bacterium isolated from a duckweed, Lemna aequinoctialis. This study quantified its growth-promoting effect on three duckweeds (L. aoukikusa, L. minor, and Spirodela polyrhiza) in sterile Hoagland solution and evaluated its usefulness in duckweed culture under non-sterile conditions. P23 promoted growth of three duckweeds in sterile Hoagland solution at low to high nutrient concentrations (1.25-10 mg NO3-N/L and 0.25-2.0 mg PO4-P/L). It increased the biomass production of L. aequinoctialis 3.8-4.3-fold, of L. minor 2.3-3.3-fold, and of S. polyrhiza 1.4-1.5-fold after 7 days compared with noninoculated controls. P23 also increased the biomass production of L. minor 2.4-fold in pond water and 1.7-fold in secondary effluent of a sewage treatment plant under non-sterile conditions at laboratory-scale experiments. P23 rescued L. minor from growth inhibition caused by microorganisms indigenous to the pond water. The results demonstrate that the use of P23 in duckweed culture can improve the efficiency of duckweed biomass production, and a positive effect of P23 on duckweed-based wastewater treatment can be assumed.

Microbial community and removal of nitrogen via the addition of a carrier in a pilot-scale duckweed-based wastewater treatment system

Carriers were added to a pilot-scale duckweed-based (Lemna japonica 0223) wastewater treatment system to immobilize and enhance microorganisms. This system and another parallel duckweed system without carriers were operated for 1.5years. The results indicated the addition of the carrier did not significantly affect the growth and composition of duckweed, the recovery of total nitrogen (TN), total phosphorus (TP) and CO2 or the removal of TP. However, it significantly improved the removal efficiency of TN and NH4+-N (by 19.97% and 15.02%, respectively). The use of 454 pyrosequencing revealed large differences of the microbial communities between the different components within a system and similarities within the same components between the two systems. The carrier biofilm had the highest bacterial diversity and relative abundance of nitrifying bacteria (3%) and denitrifying bacteria (24% of Rhodocyclaceae), which improved nitrogen removal of the system. An efficient N-removal duckweed system with enhanced microorganisms was established.

Removal of estrone, 17α-ethinylestradiol, and 17ß-estradiol in algae and duckweed-based wastewater treatment systems

Many pollutants have received significant attention due to their potential estrogenic effect and are classified as endocrine disrupting compounds (EDCs). Because of possible ecological effects and increased attention for water reuse schemes, it is important to increase our understanding of the EDC removal capacities of various wastewater treatment systems. However, there has so far been little research on the fate and behavior of EDCs in stabilization pond systems for wastewater treatment, which represent an important class of wastewater treatment systems in developing countries because of their cost-effectiveness. The aim of this work is to study the fate and behavior of EDCs in algae and duckweed ponds. Because the synthetic hormone 17alpha-ethinylestradiol (EE2) and the natural hormones estrone (E1), as well as 17beta-estradiol (E2), have been detected in effluents of sewage treatment plants and been suggested as the major compounds responsible for endocrine disruption in domestic sewage; E1, E2, and EE2 were therefore chosen as target chemicals in this current work. Both batch tests and continuous-flow tests were carried out to investigate the sorption and biodegradation of estrogens in algae and duckweed pond systems. The applied duckweed was a Lemna species. The applied algae was a mixture of pure cultures of six different algae genera, i.e., Anabaena cylindrica, Chlorococcus, Spirulina platensis, Chlorella, Scenedesmus quadricauda, and Anaebena var. Synthetic wastewater were used in all tests. The concentrations of estrogens were measured with three different enzyme-linked immunosorbent assay kits specific for E1, E2, or EE2. When the concentrations of estrogens in water samples were below the lowest quantitative analysis range (0.05 microg/l), preconcentration of the water samples were performed by means of solid phase extraction (SPE) with C18 cartridges. The 6-day batch tests show that the presence of algae or duckweed accelerated the removal of the three estrogens from the synthetic wastewater. More estrogens were removed in the tests with duckweed than in tests with algae or with wastewater. In the sorption tests, a swift sorption of the three estrogens was observed when the estrogens were contacted with duckweed or algae, while the estrogen concentrations in tap water kept unchanged during the 3-h sorption tests. The mass balances indicated that only about 5% of the estrogens were bound to the algae sediment or duckweed at the end of the 6-day tests. Results of the continuous-flow tests revealed that the algae and duckweed ponds effectively removed E1, E2, and EE2 even at nanograms per liter level. Interconversion of E1 and E2 occurred both in batch and continuous-flow tests. E2 could be readily transformed to E1, especially in the tests with algae. Different processes like sorption, biodegradation and photolytic degradation might play an important role in the removal of estrogens from the aquatic phase. The 3-h sorption tests support the importance of sorption for estrogen removal, in which a rapid initial sorption was observed over the first 2 min for E1/E2/EE2 to both duckweed and algae. In the 6-day batch tests, estrogens were sorbed by algae or duckweed during the early stage when algae and duckweed were contacted with the synthetic wastewater and the sorbed estrogens were further biodegraded by the microorganisms developed in the wastewater. The persistent estrogen concentrations in tap water, however, implied that no sorption, biodegradation, or photolytic degradation occurred in tap water under the specific experimental conditions. Under aerobic or anoxic conditions, E2 could be first oxidized to E1, which is further oxidized to unknown metabolites and finally to CO(2) and water. Under anaerobic conditions, E1 can also be reduced to E2. However, the interconversion might be much more complex especially in the tests with algae because both aerobic and anaerobic conditions occurred in these tests due to the variation of the dissolved oxygen concentration induced by the light regime. This study shows that estrogens, E1, E2, and EE2, can be effectively removed from the continuous-flow algae and duckweed ponds even when their concentrations are at nanograms per liter level. The presence of algae and duckweed accelerate the removal of estrogens from the synthetic wastewater because estrogens can be quickly sorbed on duckweed or algae. The sorbed estrogens are subsequently degraded by microorganisms, algae, or duckweed in the wastewater treatment system. E1 and E2 are interconvertible in both duckweed and algae pond systems. E2 can be readily transformed to E1, especially in the tests with algae. Based on the tests performed so far, one can conclude that both sorption and biodegradation are important to the estrogens removal from stabilization pond systems for wastewater treatment. Further research using, e.g., radioimmunoassay is needed to investigate the biodegradation pathway of estrogens in algae and duckweed ponds.

An integrated kinetic model for organic and nutrient removal by duckweed-based wastewater treatment (DUBWAT) system

This study was conducted to investigate the efficiency of duckweed ( Lemna gibba) in treating a domestic wastewater and to develop an integrated kinetic model for organic and nutrient removal by duckweed-based wastewater treatment (DUBWAT) system. Four pilot-scale DUBWAT units, made of concrete blocks, were operated under ambient conditions (temperatures 30–36 °C), different hydraulic retention times ( t), organic loading rates (OLR) and stocking densities (SD). The maximum COD, BOD 5, NH 3-N, TN and TSS removal efficiencies of 84, 88, 68, 58 and 87%, respectively, were found at optimum operating conditions of t of 10 days, OLR of 50 kgCOD/(ha-d) and SD of 0.5 kg/m 2. The nitrogen uptake rate by duckweed was found to be 0.62 g-N/(m 2-d). An integrated kinetic model consisting of t, OLR, SD and temperature was developed for the DUBWAT system and validated satisfactorily with data obtained from the literatures.

Process performance assessment of algae-based and duckweed-based wastewater treatment systems

A pilot plant experiment was carried out to assess differences in environmental conditions and treatment performance in two systems for wastewater treatment: algae-based ponds (ABP) and duckweed-based (Lemna gibba) ponds (DBP). Each system consisted of a sequence of 4 equal ponds in series and was fed with a constant flow rate of partially treated wastewater from Birzeit University. Physico-chemical parameters and the removal of organic matter, nutrients and faecal coliforms were monitored within each treatment system over a period of 12 months. The results show clear differences in the environmental conditions. In ABP significantly (P>0.05) higher pH and DO values were observed than in DBP. DBP were more efficient in removal of organic matter (BOD and TSS) than ABP. The faecal coliform reduction was higher in ABP. However, the quality of the effluent from the third and fourth duckweed pond (total retention time of 21 and 28 days) did not exceed the WHO-criteria for unrestricted irrigation during both the summer and winter period, respectively. During the summer period, the average total nitrogen was reduced more in ABP (80%) than in DBP (55%). Lower values were measured during the winter period. Seasonal nitrogen reductions of the two systems were significantly different (P>0.05). In DBP, 33% and 15% of the total nitrogen was recovered into plant biomass and removed from the system via duckweed harvesting during the summer and winter period, respectively. This study showed that there were differences in the environmental conditions and treatment efficiencies between the two systems.

Duckweed based wastewater treatment (DWWT): design guidelines for hot climates

Conventional wastewater treatment systems are expensive in either investment or running costs. On the other hand, waste stabilisation ponds may be unable to meet effluent standards for nutrients. Wastewater treatment using duckweed therefore becomes more significant as an option capable of achieving effluent standards and generating revenue from selling the duckweed. However existing duckweed based wastewater treatment (DWWT) systems have high land requirements despite being able to reduce concentrations of organic compounds and pathogens to acceptable levels. Improved guidelines for the design of DWWT are necessary to obtain a reliable and cost-effective wastewater treatment plant using duckweed. This guideline provides a DWWT design program using spreadsheets for different configurations of wastewater treatment units using duckweed. The design program developed suggests that a combination of anaerobic ponds, DWWT systems and maturation ponds can minimise land requirements and capital costs while achieving specified effluent standards. In order to achieve effluent standards, the land required is typically from 1.5 to 1.8 m2/capita (excluding associated facilities), capital costs are in the range from 7.9 to 9.7 USD/capita, with a retention time from 15 to 18 days. Income generation is dependent mainly on the social and cultural acceptability of duckweed use within the community.

Comparison between algae-based and duckweed-based wastewater treatment: differences in environmental conditions and nitrogen transformations

In laboratory-scale batch experiments, duckweed (Limna gibba)-based and algae-based wastewater containers have been monitored over 15 days in two experiments with different initial total nitrogen concentrations of 50 (experiment 1) and 100 mg-N/L (experiment 2). Clear differences in environmental conditions were observed. High dissolved oxygen (DO) concentrations were observed in the algae-based ,compared to duckweed-based, containers. In the algae-based containers the DO range was between 2.1 to 6.6 mg/L and 1.2 to 4.3 mg/L in experiment 1 and 2, respectively, whereas in the duckweed-based containers DO ranged between 1.1 to 3 mg/l and 0.5 to 2.1 mg/L. Higher pH values were measured in algae-based due to algal photosynthetic activity compared to duckweed-based containers where the duckweed mat prevented sunlight penetration and hence algal development. In algae-based containers, the pH range was 7.9 to 8.6 and 8.1 to 8.4 in experiments 1 and 2, respectively, and 7.3 to 7.5 and 7 to 7.6 in the duckweed-based containers. Depending on initial nitrogen concentrations, duckweed-based containers removed between 42%–62% of total nitrogen and between 56%–95% of Kjeldahl nitrogen from the wastewater, while algae-based containers removed between 45%–48% and 48%–58% of total nitrogen and Kjeldahl nitrogen, respectively. Nitrogen loss, probably due to denitrification and ammonia volatilisation, represents 40% of the total nitrogen content of algae-based and duckweed-based containers. However, in duckweed-based containers only 28% of N-loss was observed in containers with higher initial N-content. This study demonstrates that there were differences in environmental conditions in algae-based and duckweed-based containers, which have caused differences in nitrogen transformation mechanisms.

Advantages and limitations of duckweed-based wastewater treatment systems

In order to find a reliable, simple and cost-effective small wastewater treatment system, an experimental research programme on duckweed-based treatment systems was carried out. In spite of the profitable characteristics of duckweed (high productivity, high protein content, wide geographic distribution, control of negative impacts from conventional wastewater treatment ponds), the results obtained confirm that their extensive use in Italy seems difficult because of the high requirement of land area and the ceasing of growth in winter months (at least in Northern Italy). In temperate climates, a reasonable use of duckweed looks to be the production of good quality secondary effluents (BOD and SS removal) from small communities, especially in seasonal (summer) wastewater treatment plants. Another use is algae removal from facultative lagoon effluents. Nitrogen removal can only be reasonably obtained in duckweed-covered ponds with supplemental aeration. Because of several constraints due to the markets and to environmental regulations, harvested duckweed has to be disposed of as sewage sludge (e.g. compost or biogas production).

Advantages and limitations of duckweed-based wastewater treatment systems

In order to find a reliable. simple and cost-effective small wastewater treatment system, an experimental research programme on duckweed-based treatment systems was carried out. In spite of the profitable characteristics of duckweed (high productivity, high protein content, wide geographic distribution, control of negative impacts from conventional wastewater treatment ponds), the results obtained confirm that their extensive use in Italy seems difficult because of the high requirement of land area and the ceasing of growth in winter months (at least in Northern Italy). In temperate climates, a reasonable use of duckweed looks to be the production of good quality secondary effluents (BOD and SS removal) from small communities, especially in seasonal (summer) wastewater treatment plants. Another use is algae removal from facultative lagoon effluents. Nitrogen removal can only be reasonably obtained in duckweed-covered ponds with supplemental aeration. Because of several constraints due to the markets and to environmental regulations, harvested duckweed has to be disposed of as sewage sludge (e.g. compost or biogas production).