Biodegradation of insensitive munition (IM) formulations: IMX-101 and IMX-104 using aerobic granule technology.

A laboratory-scale aerobic granular sludge (AGS) sequencing batch bioreactor (SBR) was initiated in this study for the biodegradation of hazardous insensitive munition (IM) formulation constituents; 2,4-dinitroanisole (DNAN), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), 1-nitroguanidine (NQ), and 3-nitro-1,2,4-triazol-5-one (NTO). Efficient (bio)transformation of the influent DNAN and NTO was achieved throughout reactor operation with removal efficiencies greater than 95%. An average removal efficiency of 38.4 ± 17.5% was recorded for RDX. NQ was only slightly removed (3.96 ± 4.15%) until alkalinity was provided in the influent media, which subsequently increased the NQ removal efficiency up to an average of 65.8 ± 24.4%. Batch experiments demonstrated a competitive advantage for aerobic granular biofilms over flocculated biomass for the (bio)transformation DNAN, RDX, NTO, and NQ, as aerobic granules were capable of reductively (bio)transforming each IM compound under bulk aerobic conditions while flocculated biomass could not, thus demonstrating the contribution of inner oxygen-free zones within aerobic granules. A variety of catalytic enzymes were identified in the extracellular polymeric matrix of the AGS biomass. 16 S rDNA amplicon sequencing found Proteobacteria (27.2-81.2%) to be the most abundant phyla, with many genera associated with nutrient removal as well as genera previously described in relation to the biodegradation of explosives or related compounds.

Simultaneous reduction of perchlorate and nitrate using fast-settling anoxic sludge.

Fast-settling, anoxic sludge (FAS) was cultivated and utilized in this study to simultaneously reduce elevated levels of perchlorate and nitrate in an anaerobic sequencing batch reactor (AnSBR). Average perchlorate and nitrate removal efficiencies of 96.5 ± 8.44 % and 99.8 ± 0.32 %, respectively, were achieved from an average perchlorate and nitrate loading rate of 159 ± 101 g ClO4-/m3·d and 10.8 ± 7.25 g NO3--N/m3·d, respectively, throughout long-term operation (>500-d). Batch activity tests revealed a preferential utilization of nitrate over perchlorate, where significant perchlorate reduction inhibition occurred when nitrate was present as a competing electron acceptor under carbon-limiting conditions. Specific perchlorate and nitrate reduction rates were shown to increase as the hydraulic retention time (HRT) of the AnSBR was step-wise decreased and subsequently the perchlorate and nitrate loading rates were step-wise increased. Functional, mRNA-based expression of the nitrite reductase (nirS and nirK), nitrous oxide reductase (nosZ), perchlorate reductase subunit A (pcrA), and the chlorite dismutase (cld) genes illustrated the simultaneous activity of heterotrophic denitrification and perchlorate reduction occurring throughout a complete standard reactor operational cycle, and allowed for expression trends to be documented as the HRT of the AnSBR was reduced from 5-d to 1.25-d. Nitrous oxide (N2O) production was detected as a result of incomplete denitrification, where the largest N2O production occurred at the highest nitrate loading rates investigated in this study. Thauera species were heavily enriched at a longer HRT of 5-d, but were out-competed by Dechloromonas species as the HRT of the AnSBR was step-wise reduced to 1.25-d.

Prevalence of SARS-CoV-2 genes in water reclamation facilities: From influent to anaerobic digester.

Several treatment plants were sampled for influent, primary clarifier sludge, return activated sludge (RAS), and anaerobically digested sludge throughout nine weeks during the summer of the COVID-19 pandemic. Primary clarifier sludge had a significantly higher number of SARS-CoV-2 gene copy number per liter (GC/L) than other sludge samples, within a range from 1.0 × 105 to 1.0 × 106 GC/L. Gene copy numbers in raw influent significantly correlated with gene copy numbers in RAS in Silver Creek (p-value = 0.007, R2 = 0.681) and East Canyon (p-value = 0.009, R2 = 0.775) WRFs; both of which lack primary clarifiers or industrial pretreatment processes. This data indicates that SARS-CoV-2 gene copies tend to partition into primary clarifier sludges, at which point a significant portion of them are removed through sedimentation. Furthermore, it was found that East Canyon WRF gene copy numbers in influent were a significant predictor of daily cases (p-value = 0.0322, R2 = 0.561), and gene copy numbers in RAS were a significant predictor of weekly cases (p-value = 0.0597, R2 = 0.449). However, gene copy numbers found in primary sludge samples from other plants significantly predicted the number of COVID-19 cases for the following week (t = 2.279) and the week after that (t = 2.122) respectively. These data indicate that SARS-CoV-2 extracted from WRF biosolids may better suit epidemiological monitoring that exhibits a time lag. It also supports the observation that primary sludge removes a significant portion of SARS-CoV-2 marker genes. In its absence, RAS can also be used to predict the number of COVID-19 cases due to direct flow through from influent. This research represents the first of its kind to thoroughly examine SARS-CoV-2 gene copy numbers in biosolids throughout the wastewater treatment process and the relationship between primary, return activated, and anaerobically digested sludge and reported positive COVID-19 cases.

Per and poly-fluoroalkyl substances (PFAS) as a contaminant of emerging concern in surface water: A transboundary review of their occurrences and toxicity effects.

Per and poly-fluoroalkyl substances (PFAS) have been recognized as contaminants of emerging concerns by the United States Environmental Protection Agency (US EPA) due to their environmental impact. Several advisory guidelines were proposed worldwide aimed at limiting their occurrences in the aquatic environments, especially for perfluorooctane sulfonic acid (PFOS) and perfluorooctanoic acid (PFOA). This review paper aims to provide a holistic review in the emerging area of PFAS research by summarizing the spatiotemporal variations in PFAS concentrations in surface water systems globally, highlighting the possible trends of occurrences of PFAS, and presenting potential human health impacts as a result of PFAS exposure through surface water matrices. From the data analysis in this study, occurrences of PFOA and PFOS in many surface water matrices were observed to be several folds higher than the US EPA health advisory level of 70 ng/L for lifetime exposure from drinking water. Direct discharge and atmospheric deposition were identified as primary sources of PFAS in surface water and cryosphere, respectively. While global efforts focused on limiting usages of long-chain PFAS such as PFOS and PFOA, the practices of using short-chain PFAS such as perfluorobutanoic acid (PFBA) and perfluorobutane sulfonic acid (PFBS) and PFAS alternatives increased substantially. These compounds are also potentially associated with adverse impacts on human health, animals and biota.

An overview of fungal pretreatment processes for anaerobic digestion: Applications, bottlenecks and future needs.

Lignin modifying or extracellular enzymes secreted by the white rot fungi have the ability of degrading wide range of lignocellulosic substrates and organic pollutants. Lignocellulosic biomass, despite being a renewable source of energy, is difficult to hydrolyse (hydrolysis being rate-limiting stage in anaerobic digestion process). Various pre-treatment techniques like physical, chemical, thermo-chemical and biological to enhance the accessibility of microbes to carbohydrates have been studied. Recently, usage of white- rot fungi in a biological pre-treatment technique have received renewed interest due to its low cost and eco-friendly nature. This review deals with: a) lignocellulosic biomass recalcitrance, b) various pre-treatment techniques and its economic feasibility, c) delignification and hydrolysis mechanism using white-rot fungi, d) factors controlling white-rot fungi pre- treatment process, and e) improvement in methane production through solid-state anaerobic digestion of white-rot fungi pre-treated lignocellulosic biomass. Finally a future perspective is also included.

Cyanotoxin impact on microbial-mediated nitrogen transformations at the interface of sediment-water column in surface water bodies.

Harmful cyanobacterial blooms produce lethal toxins in many aquatic ecosystems experiencing eutrophication. This manuscript presents results on the effects of cyanotoxins on the aerobic microbial communities residing at the interface of sediments and water columns with the ammonia-oxidizing bacteria (AOB) as the model microbial community. Microcystin-LR (MC-LR), a heavily researched cyanotoxin variant, was used as the model cyanotoxin. To measure cyanotoxin influence on the activity of nitrifying microbial communities, an enriched culture of AOBs collected from an ongoing partial nitrification-nitritation reactor was examined for its exposure to 1, 5 and 10 µg/L of MC-LR. The nitritation kinetics experiment demonstrated MC-LR's ability at 1, 5, and 10 µg/L concentrations to prevent ammonium oxidation with statistically significant differences in nitritation rates between the blanks and spiked samples (One-way ANOVA, p < 0.05). Significantly decreased dissolved oxygen (DO) consumption during oxygen update batch tests demonstrated toxin's influence on AOB's oxidizing capabilities when exposed to even lower concentrations of 0.75, 0.5, and 0.25 µg/L of MC-LR in a separate set of experiments. Based on competitive kinetics, the MC-LR inhibition coefficient-the concentration needed to produce half-maximum inhibition of the mixed community AOBs was determined to be 0.083 µg/L. The stress tests proved the recovery of nitritation to some extent at lower MC-LR concentrations (1 and 5 µg/L), but significant irreversible inhibition was recorded when the AOB population was exposed to 10 µg/L MC-LR. The comparisons of amoA gene expressions corresponded well with nitrifying kinetics. All concentrations of MC-LR spiking were determined to produce a discernible impact on the AOB nitritation rate by either destroying the bacterial cell or immediately inhibiting the amoA gene expression.

Nitrogen management in landfill leachate using single-stage anammox process-illustrating key nitrogen pathways under an ecogenomics framework.

Simultaneous functional gene expressions using mRNA, rate measurements, and biochemical analysis proved the consistent contribution of ammonia oxidizers, heterotrophic denitrifiers, and anammox bacteria in a single-stage attached growth partial nitritation/anammox system for nitrogen management in landfill leachate. Average removal efficiencies of ammonia-nitrogen, total inorganic nitrogen, and COD were 94%, 88%, and 26%, respectively, in the reactor. Off-gas N2O fluxes increased at relatively higher dissolved oxygen. Batch activity tests revealed the occurrence of significant anammox activity even in the presence of high concentrations of organic carbon in the influent. mRNA based functional expressions of nitrite reductase (nirK and nirS) and hydrazine synthase (hzsA) suggested simultaneous active heterotrophic denitrification and anammox, respectively. 16S rRNA amplicon sequencing revealed Proteobacteria (36-56%), Planctomycetes (10-31%), and Bacteroidetes (6-39%) as dominant phyla in the reactor. Candidatus brocadia was observed as the most abundant genus representing anammox community.

Response of an aerobic granular and conventional flocculated reactors against changing feed composition from simple composition to more complex.

This research evaluated the effect of changing feed composition on the performances of a conventional activated sludge (CAS) and an aerobic granular sludge (AGS) reactor operated simultaneously. Both reactors were initially fed with 100% synthetic feed. In a stepwise manner, the feed composition was slowly changed to real primary effluent collected from a local wastewater treatment plant. After an initial stabilization period, both reactors could achieve more than 90% NH4+-N removal. However, PO43--P removal eventually reached to a maximum of 92% in the AGS and 88% in the CAS. COD removal in both reactors was least affected, with the lowest percent removal of 81 ± 3% achieved in AGS and 62 ± 4% in CAS respectively when fed with 100% real wastewater. Despite granule breakage the AGS reactor was able to remove the pollutants (COD, N, P). The abundance of Candidatus Accumulibacter, a polyphosphate accumulating organism, in the AGS system increased over the operational phases: II (6.2%), III (10.32%), and IV (11.9%). While in CAS, it increased from phase I to phase II (12.6%), but decreased in phase III to 9.9%. Genus-based classification revealed a successive increase in the relative abundance of Nitrospira to 11.05% during Phase III and 10.3% during Phase IV in the AGS. In contrast with its presence in the CAS, which was, 3.4% during Phase III and 9.5% during Phase IV.

Integrated leachate management approach incorporating nutrient recovery and removal.

This manuscript presents an integrated management scheme for leachate which employed struvite precipitation to recover ammonia nitrogen and phosphorus, aerobic granular sludge process for carbon oxidation (in the form of BOD and sCOD) and single stage anaerobic ammonia oxidation (ANAMMOX) for nitrogen management. The influent fed to the integrated treatment scheme was a mixture of anaerobic digester centrate and real leachate in 4:1 ratio. Almost 77% recovery of phosphorus and 25% removal of NH4+-N were accomplished through struvite precipitation at an optimum pH of 9. High pH contributed to free ammonia loss during struvite precipitation experiments. In the aerobic granular sludge reactor overall, BOD5, COD and NH4+-N removal percentages were 74%, 45% and 35% and in the PN/A reactor, overall 35% removal of total inorganic nitrogen (TIN) was observed. More than 80% BOD removal was recorded in the granular reactor with soluble COD (sCOD) removal fluctuating between 28 and 57% depending on the operational phase. High-throughput amplicon sequencing of 16S rRNA gene targeting V4 region revealed a dominance of phylum Planctomycetes, in the PN/A reactor system. Presence of Rhodobacteraceae, Xanthomonadaceae, Flavobacteriaceae in the granular biomass confirmed the defined redox zones inside mature granules indicating simultaneous removal of nitrogen (N) and organics in aerobic granular sludge technology. Exposing the synthetically cultured aerobic granules directly to the mixture of leachate and centrate unveiled an alteration in physical characteristics of granules; however, reactor operational data and microbial community analysis ascertain the effectiveness of the treatment scheme treating two urban waste-streams.