Advantageous microbial community development and improved performance of pilot-scale field systems treating high-risk acid mine drainage with crab shell.

Microbial communities are crucial to the effectiveness and stability of bioremediation systems treating acid mine drainage (AMD); however, little research has addressed how they correlate to system performance under changing environmental conditions. In this study, 16S rRNA gene sequencing and quantitative PCR (qPCR) were used to characterize microbial communities within different substrate combinations of crab shell (CS) and spent mushroom compost (SMC) and their association with chemical performance in pilot-scale vertical flow ponds (VFPs) treating high risk AMD in central Pennsylvania over 643 days of operation. As compared to a control containing SMC, VFPs containing CS sustained higher alkalinity, higher sulfate-reducing rates, and more thorough metals removal (>90% for Fe and Al, >50% for Mn and Zn). Correspondingly, CS VFPs supported the growth of microorganisms in key functional groups at increasing abundance and diversity over time, especially more diverse sulfate-reducing bacteria. Through changing seasonal and operational conditions over almost two years, the relative abundance of the core phyla shifted in all reactors, but the smallest changes in functional gene copies were observed in VFPs containing CS. These results suggest that the high diversity and stability of microbial communities associated with CS are consistent with effective AMD treatment.

Crab shell amendments enhance the abundance and diversity of key microbial groups in sulfate-reducing columns treating acid mine drainage.

Substrate amendments composed of crab shell (CS) waste materials have been shown to significantly improve the longevity and performance of acid mine drainage (AMD) treatment systems containing spent mushroom compost (SMC), yet the development of key microbial populations within these systems has not been investigated. To better understand the effects of CS on microbial dynamics in these systems, clone libraries and real-time quantitative PCR (qPCR) were performed on materials from a laboratory-scale AMD treatment system containing SMC and 0 to 100% CS substrate after receiving a continuous flow of AMD for 148 days (428 pore volumes). The proportion of CS in the substrate positively correlated with the diversity of sulfate-reducing bacteria (SRB) and archaeal clones, but negatively correlated with fungal diversity. CS also impacted microbial community structure, as revealed in Unifrac significance and principal coordinate analysis tests. The column containing 100% CS substrate supported 7 different genera of SRB-the most ever observed in an AMD treatment system. Moreover, the copy numbers of functional genes representing fermenters, sulfate reducers, and chitin degraders increased with increasing proportions of CS. These observations agree well with the chemical performance data, further validating that by supporting more abundant key microbial groups, chitinous substrates may provide benefits for improving both the longevity and performance of AMD treatment systems, and may provide similar benefits for the treatment of other environmental contaminants that are amenable to anaerobic bioremediation.Key points• Crab shell improves the longevity and performance of acid mine drainage treatment.• The diversity of sulfate-reducing bacteria is enhanced with crab shell amendments.• Crab shell supports more abundant key microbial groups than spent mushroom compost.

Anaerobic bioprocessing of wastewater-derived duckweed: Maximizing product yields in a biorefinery value cascade.

This study integrated the sugar and carboxylate platforms to enhance duckweed processing in biorefineries. Two or three bioprocesses (ethanol fermentation, acidogenic digestion, and methanogenic digestion) were sequentially integrated to maximize the carbon-to-carbon conversion of wastewater-derived duckweed into bioproducts, through a series of laboratory-scale experiments. Reactors were fed either raw (dried), liquid-hot-water-pretreated, or enzymatically-saccharified duckweed. Subsequently, the target bioproduct was separated from the reactor liquor and the residues further processed. The total bioproduct carbon yield of 0.69 ±â€¯0.07 g per gram of duckweed-C was obtained by sequential acidogenic and methanogenic digestion. Three sequential bioprocesses revealed nearly as high yields (0.66 ±â€¯0.08 g of bioproduct-C per duckweed-C), but caused more gaseous carbon (dioxide) loss. For this three-stage value cascade, yields of each process in conventional units were: 0.186 ±â€¯0.001 g ethanol/g duckweed; 611 ±â€¯64 mg volatile fatty acids as acetic acid/g VS; and 434 ±â€¯0.2 ml methane/g VS.

Duckweed as an Agricultural Amendment: Nitrogen Mineralization, Leaching, and Sorghum Uptake.

Excessive N and P in surface waters can promote eutrophication (algae-dominated, low-O waters), which decreases water quality and aquatic life. Duckweed (Lemnaceae), a floating aquatic plant, rapidly absorbs N and P from water and its composition shows strong potential as a soil amendment. Therefore, it may be used to transfer N and P from eutrophic water bodies to agricultural fields. In this work, dried duckweed was incorporated into agricultural soil in microcosm, column, and field tests to evaluate biological N cycling, nutrient retention, and crop yield compared with compost, diammonium phosphate (DAP), and an amendment-free control. In microcosm tests, 25 ± 13% of duckweed N was mineralized, providing on average less mineral N than DAP (107 ± 21%), but more than compost (11 ± 12%). In columns, duckweed treatments leached only 2% of the N added, significantly less than DAP, which leached 60% of its N. Compared with the control, DAP leached significantly more phosphate (78%), whereas duckweed and compost treatments leached less (56 and 27%, respectively). Crop yield, as well as runoff N and P, were measured in field tests growing forage sorghum [ (L.) Moench.]. Although less total N was applied to duckweed plots than to DAP plots (75 vs. 130 kg ha, respectively), duckweed was found to retain 30% more total mineral N in a tilled agricultural field than DAP, while supporting a comparable yield. These tests indicate that duckweed may provide a sustainable source of N and P for agriculture.

Trace organic contaminant removal in six full-scale integrated fixed-film activated sludge (IFAS) systems treating municipal wastewater.

Trace organic contaminants (TrOCs) often pass through conventional activated sludge wastewater treatment plants (CAS-WWTPs) and are discharged into surface waters, where they can threaten aquatic ecosystems and human health, largely due to the hormone disrupting effects of certain TrOCs. The integrated fixed-film activated sludge (IFAS) process is a cost-effective means of upgrading CAS-WWTPs by adding free-floating carrier media, which promotes biofilm formation in the well-mixed suspended growth reactors, providing a potential niche for slow-growing microorganisms. Although IFAS upgrades are typically aimed at enhancing nutrient removal, limited bench- and pilot-scale data indicate that TrOC removal may also be improved. However, only limited reports which focus on a small number of compounds in individual full-scale IFAS-WWTPs have been published to date, and no data is available regarding the removal of estrogenic activity in full-scale IFAS-WWTPs. In this study, six full-scale IFAS-WWTPs were surveyed to quantify TrOC and estrogenic activity removal. Twenty-four hour composite samples of secondary influent and effluent (pre-disinfection) were analyzed for total suspended solids (TSS), chemical oxygen demand (COD), ammonia, total nitrogen (TN), total phosphorus (TP), estrogenic activity, and 98 TrOCs. The biomass distribution between the suspended growth phase (i.e. mixed liquor) and IFAS media was also assessed. All IFAS-WWTPs performed well in terms of TSS, COD, and ammonia removal. TN removal varied in accordance with nitrate removal. Total solids per liter of wetted reactor volume ranged from 2.5 to 7.6 g, with 40-60% attached to media. TrOCs with no detection (17) and those with high median removal (23, ≥90% average removal) were observed. Other TrOCs had lower and more variable removal efficiencies. Qualitative comparison with CAS literature shows potentially higher IFAS removal efficiencies for a number of compounds including several which have been previously indicated in bench- or pilot-scale studies (atenolol, diclofenac, gemfibrozil, DEET, 4-nonylphenol, and 4-tert-octylphenol), as well as the chlorinated flame retardants TCIPP and TDCIPP. Effluent estrogenic activity was found to be similar to that reported for full-scale CAS-WWTPs. These results provide the first survey of multiple full-scale IFAS-WWTPs employing mobile plastic carrier media in terms of basic chemical endpoints (removal of ammonia, TN, TP, and COD), the distribution of solids within the systems, and the removal of TrOCs and estrogenic activity.

Effect of pH and temperature on microbial community structure and carboxylic acid yield during the acidogenic digestion of duckweed.

BACKGROUND: Duckweeds (Lemnaceae) are efficient aquatic plants for wastewater treatment due to their high nutrient-uptake capabilities and resilience to severe environmental conditions. Combined with their rapid growth rates, high starch, and low lignin contents, duckweeds have also gained popularity as a biofuel feedstock for thermochemical conversion and alcohol fermentation. However, studies on the acidogenic anaerobic digestion of duckweed into carboxylic acids, another group of chemicals which are precursors of higher-value chemicals and biofuels, are lacking. In this study, a series of laboratory batch experiments were performed to determine the favorable operating conditions (i.e., temperature and pH) to maximize carboxylic acid production from wastewater-derived duckweed during acidogenic digestion. Batch reactors with 25 g/l solid loading were operated anaerobically for 21 days under mesophilic (35 °C) or thermophilic (55 °C) conditions at an acidic (5.3) or basic (9.2) pH. At the conclusion of the experiment, the dominant microbial communities under various operating conditions were assessed using high-throughput sequencing. RESULTS: The highest duckweed-carboxylic acid conversion of 388 ± 28 mg acetic acid equivalent per gram volatile solids was observed under mesophilic and basic conditions, with an average production rate of 0.59 g/l/day. This result is comparable to those reported for acidogenic digestion of other organics such as food waste. The superior performance observed under these conditions was attributed to both chemical treatment and microbial bioconversion. Hydrogen recovery was only observed under acidic thermophilic conditions, as 23.5 ± 0.5 ml/g of duckweed volatile solids added. More than temperature, pH controlled the overall structure of the microbial communities. For instance, differentially abundant enrichments of Veillonellaceae acidaminococcus were observed in acidic samples, whereas enrichments of Clostridiaceae alkaliphilus were found in the basic samples. Acidic mesophilic conditions were found to enrich acetoclastic methanogenic populations over processing times longer than 10 days. CONCLUSIONS: Operating conditions have a significant effect on the yield and composition of the end products resulting from acidogenic digestion of duckweed. Wastewater-derived duckweed is a technically feasible alternative feedstock for the production of advanced biofuel precursors; however, techno-economic analysis is needed to determine integrated full-scale system feasibility and economic viability.

In situ bioremediation of perchlorate in vadose zone soil using gaseous electron donors: microcosm treatability study.

A series of microcosm experiments were performed to determine the effectiveness of various gaseous electron donors (including hydrogen, 1-hexene, ethyl acetate, and liquefied petroleum gas [LPG]) for supporting biological perchlorate reduction under different electron donor concentrations and soil moistures. Under high soil moisture (16% w/w) conditions, complete or partial perchlorate degradation was achieved with all of the tested electron donors, except for ethyl acetate. Hydrogen was the most promising of the tested electron donors, achieving complete perchlorate degradation with first-order rate constants ranging from 0.13 to 0.20 day(-1) and reducing concentrations to non-detectable levels within 35 to 42 days. The LPG and 1-hexene each promoted partial perchlorate reduction, with average first-order rate constants of 0.05 and 0.11 day(-1), respectively. Although significant perchlorate reduction was observed with as little as 13% moisture, the moisture content for complete perchlorate degradation in this particular soil was determined to be 17%.

Efficient metal removal and neutralization of acid mine drainage by crab-shell chitin under batch and continuous-flow conditions.

Crab-shell chitin was evaluated as a multifunctional substrate for treating acid mine drainage (AMD) in both batch-microcosms and continuous-flow column tests. In microcosms, crab-shell chitin was able to treat AMD from three different sites with similar results: pH increased from 3.5 to approximately 7.5 within 2 days; alkalinity increased at a rate of 37.9+/-2.2 mg CaCO(3)/L day; and sulfate was reduced at a rate of -13.6+/-2.6 mg SO(4)(2-)/L day. In columns, a hydraulic retention time of 11.2 h was enough to raise the pH from 3.5 to approximately 7.5. Alkalinity increased at a rate of 50+/-20 mg CaCO(3)/day, and lasted throughout the duration of the test (125 days, 268 pore volumes (PV)) without showing signs of exhaustion. Metals (Al, Fe, and Mn) were completely removed for 171 PV, and geochemical modeling indicates that they likely precipitated as insoluble hydr(oxides), sulfides, and carbonates. Manganese and iron breakthroughs occurred after 174 and 234 PV, respectively, whereas aluminum breakthrough was never observed. These results demonstrate for the first time that crab-shell chitin can completely remove metals and neutralize the pH of AMD under continuous-flow conditions.

The use of crab-shell chitin for biological denitrification: batch and column tests.

Crab-shell chitin (SC-20) was evaluated for its ability to enhance biological denitrification in bench-scale tests. In the presence of SC-20, highly reducing conditions were generated, supporting both denitrification and sulfate reduction of aerated water. Rapid degradation of protein in SC-20 was observed to cause an initial high release of ammonium and carbon, while a slower, continuous release of calcium carbonate from the crab shell maintained the pH near 9 throughout the tests. In batch tests, denitrification rates of 2.4+/-0.2 mg N/L-d were obtained. Columns receiving a continuous nitrate load of 24.5 mg N/L-d sustained complete denitrification for an average of 149 d (250 pore volumes). The denitrification rates and longevity of SC-20 chitin are comparable to, or better than, those previously reported for other polymeric substrates. This, in addition to its particle size, non-swelling nature, and ease of delivery in slurry form make SC-20 an attractive electron donor source for groundwater bio-denitrification.

Substrate-enhanced microbial fuel cells for improved remote power generation from sediment-based systems.

A sediment microbial fuel cell (MFC) produces electricity through the bacterial oxidation of organic matter contained in the sediment. The power density is limited, however, due in part to the low organic matter content of most marine sediments. To increase power generation from these devices, particulate substrates were added to the anode compartment. Three materials were tested: two commercially available chitin products differing in particle size and biodegradability (Chitin 20 and Chitin 80) and cellulose powder. Maximum power densities using chitin in this substrate-enhanced sediment MFC (SEM) were 76 +/- 25 and 84 +/- 10 mW/m2 (normalized to cathode projected surface area) for Chitin 20 and Chitin 80, respectively, versus less than 2 mW/m2 for an unamended control. Power generation over a 10 day period averaged 64 +/- 27 mW/ m2 (Chitin 20) and 76 +/- 15 mW/m2 (Chitin 80). With cellulose, a similar maximum power was initially generated (83 +/- 3 mW/m2), but power rapidly decreased after only 20 h. Maximum power densities over the next 5 days varied substantially among replicate cellulose-fed reactors, ranging from 29 +/- 12 to 62 +/- 23 mW/m2. These results suggest a new approach to power generation in remote areas based on the use of particulate substrates. While the longevity of the SEM was relatively short in these studies, it is possible to increase operation times by controlling particle size, mass, and type of material needed to achieve desired power levels that could theoretically be sustained over periods of years or even decades.

Chitin and corncobs as electron donor sources for the reductive dechlorination of tetrachloroethene.

Chitin, corncobs, and a mixture of chitin and corncobs were tested as potential electron donor sources for stimulating the reductive dechlorination of tetrachloroethene (PCE). Semi-batch, sand-packed columns were used to evaluate the donors with aerobic and anaerobic groundwaters containing varying degrees of alkalinity. In all experiments, acetate and butyrate were the dominant fatty acids produced, although propionate, valerate, formate, and succinate were also detected. From a multivariable regression analysis on the data, the presence of chitin, limestone, and dechlorinating culture inoculum were determined to be the most positive predictors of dechlorination activity. Chitin fermentation products supported the degradation of PCE to trichloroethene (TCE), cis-1,2-dichloroethene (DCE), and vinyl chloride (VC), even in columns containing PCE DNAPL, whereas dechlorination activity was not observed in any of the columns containing corncobs alone. The longevity and efficiency of chitin as an electron donor source demonstrates its potential usefulness for passive, in situ field applications.

Continuous steady-state method using tenax for delivering tetrachloroethene to chloro-respiring bacteria.

Tenax-TA, a solid-phase sorbent, was used as an alternative to hexadecane for continuous delivery of tetrachloroethene (PCE) to Desulfuromonas strain BB1, a chloro-respiring microorganism. In both batch and bioreactor configurations, Tenax not only maintained low, steady-state concentrations of PCE in an active culture for several months but also adsorbed the product of dechlorination, cis-1,2-dichloroethene, before it approached toxic levels.