Uptake and translocation of brominated flame retardants in tomato plants (Solanum lycopersicum L.): Results from a standard soil-based biotest.

The uptake and translocation of four polybrominated diphenyl ethers (PBDEs) and four novel brominated flame retardants (NBFRs) in tomato plants (Solanum lycopersicum L.) were investigated via the RHIZOtest, a standard soil-based biotest, optimized for organic compounds. Tomato plants were exposed to soil samples spiked with 0 (i.e. control), 5.00 or 50.00 ng g-1dw of each compound. Compared of those of the control, exposure to increasing spiking concentrations resulted in average reductions of 13% and 26% (w/w) in tomato plant biomass. Higher concentrations of NBFRs were analyzed both in roots, ranging from 0.23 to 8.01 ng g-1dw for PBDEs and from 1.25 to 18.51 ng g-1dw for NBFRs, and in shoots, ranging from 0.09 to 5.58 ng g-1dw and from 0.47 to 7.78 ng g-1dw for PBDEs and NBFRs, respectively. This corresponded to an average soil uptake of 5% for PBDEs and 9% for NBFRs at the lower soil-spiking level, and 3% for PBDEs and 6% for NBFRs at the higher soil spiking level. Consequently, among both initial spiking levels, the soil-root concentration factor (RCF) values were lower on average for PBDEs (0.13 ± 0.05 g dw soil g-1dw roots) than for NBFRs (0.33 ± 0.16 g dw soil g-1dw roots). Conversely, nondifferent values of the root-shoot transfer factor (TF) were calculated for both PBDEs (0.54 ± 0.13 g dw roots g-1dw shoots) and NBFRs (0.49 ± 0.24 g dw roots g-1dw shoots). The differences and similarities reported in the RCF and TF between and within the two groups of compounds can be explained by their properties. The calculated RCF and TF values of the PBDEs exhibited a decreasing trend as the number of bromine atoms increased. Additionally, a robust negative linear correlation was observed between RCF values and the respective logKow values for the PBDEs, at both soil-spiking levels. The root uptake of NBFRs exhibited a negative correlation with their hydrophobicity; however, this was not observed in the context of root-to-shoot transfer. The presence of a second aromatic ring appears to be the key factor influencing the observed variations in NBFRs, with biphenyl NBFRs (BTBPE and DBDPE) characterized by lower uptake and reduced translocation potential than monophenyl PBEB and HBB. Understanding the transfer of these compounds to crops, especially near plastic recycling waste sites, is crucial for understanding the risks of their potential inclusion in the human food chain.

Environmental pitfalls and associated human health risks and ecological impacts from landfill leachate contaminants: Current evidence, recommended interventions and future directions.

The improper management of solid waste, particularly the dumping of untreated municipal solid waste, poses a growing global challenge in both developed and developing nations. The generation of leachate is one of the significant issues that arise from this practice, and it can have harmful impacts on both the environment and public health. This paper presents an overview of the primary waste types that generate landfill leachate and their characteristics. This includes examining the distribution of waste types in landfills globally and how they have changed over time, which can provide valuable insights into potential pollutants in a given area and their trends. With a lack of specific regulations and growing concerns regarding environmental and health impacts, the paper also focuses on emerging contaminants. Furthermore, the environmental and ecological impacts of leachate, along with associated health risks, are analyzed. The potential applications of landfill leachate, suggested interventions and future directions are also discussed in the manuscript. Finally, this work addresses future research directions in landfill leachate studies, with attention, for the first time to the potentialities that artificial intelligence can offer for landfill leachate management, studies, and applications.

Fragrance materials affect life history parameters and gene expression in Daphnia magna: An emerging issue for freshwater ecosystems.

A chronic toxicity test (21 d exposure) with the model organism Daphnia magna was performed to study the single-compound and combined effects of four fragrance materials (FMs), including musk xylene (MX), Celestolide™ (ADBI), Galaxolide™ (HHCB), and ethylene brassylate (MT). Furthermore, the transcriptional responses of ten target genes related to detoxification, molting and reproduction (DHR96, P-gp, CYP360A8, GST, CYP314, EcRb, Vtg, CAT, GPX, and GCLC) were determined by performing a quantitative real-time polymerase chain reaction (qRT‒PCR) after juvenile D. magna was exposed for 48 h. The results showed that MX, ADBI and HHCB affected development and reproduction after chronic exposure at a concentration of 10 µg L-1. Conversely, MT did not affect reproduction, growth or molting during the 21 d exposure. In juvenile D. magna, gene expression was significantly altered by ADBI (DHR96, CYP260A8, and GCLC) and MX (DHR96, CYP360A8, EcRb, Vtg, CYP314, and GCLC) but not by HHCB. These results suggest that compared to biochemical measures, conventional biological endpoints provide more informative data regarding the effects of this FM. Compared to single substances in the chronic test, the mixture of the four FMs showed effects at lower concentrations and increased gene expression for EcRb and CYP314 during juvenile exposure, indicating a possible additive or synergistic effect of the four FMs compared to single compound exposure.

SARS-CoV-2 in Atmospheric Particulate Matter: An Experimental Survey in the Province of Venice in Northern Italy.

Analysis of atmospheric particulate matter (PM) has been proposed for the environmental surveillance of SARS-CoV-2. The aim of this study was to increase the current knowledge about the occurrence of SARS-CoV-2 in atmospheric PM, introduce a dedicated sampling method, and perform a simultaneous assessment of human seasonal coronavirus 229E. Thirty-two PM samples were collected on quartz fiber filters and six on Teflon using a low- and high-volumetric rate sampler, respectively, adopting a novel procedure for optimized virus detection. Sampling was performed at different sites in the Venice area (Italy) between 21 February and 8 March 2020 (n = 16) and between 27 October and 25 November 2020 (n = 22). A total of 14 samples were positive for Coronavirus 229E, 11 of which were collected in October-November 2020 (11/22; positivity rate 50%) and 3 in February-March 2020 (3/16 samples, 19%). A total of 24 samples (63%) were positive for SARS-CoV-2. Most of the positive filters were collected in October-November 2020 (19/22; positivity rate, 86%), whereas the remaining five were collected in February-March 2020 at two distinct sites (5/16, 31%). These findings suggest that outdoor PM analysis could be a promising tool for environmental surveillance. The results report a low concentration of SARS-CoV-2 in outdoor air, supporting a scarce contribution to the spread of infection.

Maize plant (Zea mays) uptake of organophosphorus and novel brominated flame retardants from hydroponic cultures.

The root uptake and root-shoot translocation of seven organophosphorus flame retardants (OPFRs) and four novel brominated flame retardants (NBFRs) were assessed in this investigation using hydroponic grown maize plants (Zea mays). Three initial liquid concentrations for each considered compound were examined (i.e., 0.3 µg L-1, 3 µg L-1, 30 µg L-1). The results indicated that the 30 µg L-1 treatments were phytotoxic, as they resulted in a significant decrease in shoot dry weight. Plant-driven removal of the tested FRs decreased with the increasing initial spiking level and were reportedly higher for the NBFRs (range 42%-10%) than OPFRs (range 19%-7%). All the considered FRs were measured in the roots (range 0.020-6.123 µg g-1 dry weight -DW-) and shoots (range 0.012-1.364 µg g-1 DW) of the tested plants, confirming that there was uptake. Linear relationships were identified between the chemical concentrations in the plant parts and the tested hydroponic concentrations. Root concentration factors were positively correlated with the specific lipophilicity (i.e., logKow) of the tested FRs and were determined to be higher for the NBFRs than the OPFRs. The NBFRs had a higher root uptake rate than the OPFRs, and this trend was more significant with the increasing treatment concentrations. Shoot/root concentration factors were found to be lower than the unity value for 10 of the 11 tested compounds. These results can be related to the specific molecular configurations and the occurrence of different functional groups in the tested compounds. The results will help to improve risk assessment procedures and fine tune our understanding of human receptor responses to the ingestion of maize crops grown on agricultural sites irrigated with water contaminated by FRs.

Agricultural application of digestates derived from agricultural and municipal organic wastes: a health risk-assessment for heavy metals.

A Human-health Risk Assessment was performed for an agricultural site in North-East Italy undergone digestate application to (i) check the compliance of digestate land spreading with the Italian and European regulations on contaminated agricultural soils and (ii) evaluate how resulting risk estimations can be influenced by the applied modeling assumptions. The assessment estimated the risk related to adults and children intake of Heavy Metals (HM) contained in crops at concentrations estimated by a soil-plant transfer model based on the substance-specific soil-water partition coefficients. Eight different scenarios were investigated, according to different digestate type (from biowaste and agro-industrial byproducts), digestate application techniques and soil background concentrations. Non-risky situations resulted in all scenarios involving digestate application. The totality of calculated non-carcinogenic Hazard Indexes (HI) and carcinogenic total risk (RTOTC) resulted below 0.02 and 3E10-9, respectively. In contrast with the definition, non-carcinogenic risks were associated with the considered soil background concentrations, with HI s up to 1.7 for child receptors, while carcinogenic risk was calculated below the concern threshold (i.e., RTOTC < 10-5). Accordingly, this study highlighted (i) non-concerning situations related with lawful application of digestates and (ii) the need to improve the modeling of bioavailability to plant of HMs background content of soil.

A review of the presence of SARS-CoV-2 RNA in wastewater and airborne particulates and its use for virus spreading surveillance.

According to the WHO, on October 16, 2020, the spreading of the SARS-CoV-2, responsible for the COVID-19 pandemic, reached 235 countries and territories, and resulting in more than 39 million confirmed cases and 1.09 million deaths globally. Monitoring of the virus outbreak is one of the main activities pursued to limiting the number of infected people and decreasing the number of deaths that have caused high pressure on the health care, social, and economic systems of different countries. Wastewater based epidemiology (WBE), already adopted for the surveillance of life style and health conditions of communities, shows interesting features for the monitoring of the COVID-19 diffusion. Together with wastewater, the analysis of airborne particles has been recently suggested as another useful tool for detecting the presence of SARS-CoV-2 in given areas. The present review reports the status of research currently performed concerning the monitoring of SARS-CoV-2 spreading by WBE and airborne particles. The former have been more investigated, whereas the latter is still at a very early stage, with a limited number of very recent studies. Nevertheless, the main results highlights in both cases necessitate more research activity for better understating and defining the biomarkers and the related sampling and analysis procedures to be used for this important aim.

Organic waste biorefineries: Looking towards implementation.

The concept of biorefinery expands the possibilities to extract value from organic matter in form of either bespoke crops or organic waste. The viability of biorefinery schemes depends on the recovery of higher-value chemicals with potential for a wide distribution and an untapped marketability. The feasibility of biorefining organic waste is enhanced by the fact that the biorefinery will typically receive a waste management fee for accepting organic waste. The development and implementation of waste biorefinery concepts can open up a wide array of possibilities to shift waste management towards higher sustainability. However, barriers encompassing environmental, technical, economic, logistic, social and legislative aspects need to be overcome. For instance, waste biorefineries are likely to be complex systems due to the variability, heterogeneity and low purity of waste materials as opposed to dedicated biomasses. This article discusses the drivers that can make the biorefinery concept applicable to waste management and the possibilities for its development to full scale. Technological, strategic and market constraints affect the successful implementations of these systems. Fluctuations in waste characteristics, the level of contamination in the organic waste fraction, the proximity of the organic waste resource, the markets for the biorefinery products, the potential for integration with other industrial processes and disposal of final residues are all critical aspects requiring detailed analysis. Furthermore, interventions from policy makers are necessary to foster sustainable bio-based solutions for waste management.

Effects of woody biochar on dry thermophilic anaerobic digestion of organic fraction of municipal solid waste.

This study presents the results of semi-pilot scale anaerobic digestion tests conducted under dry thermophilic conditions with the addition of biochar (6% on fresh mass basis of inoculum), derived from an industrial gasification plant, for determining biogas and biomethane production from organic fraction of municipal solid waste. By using two types of inocula (from a full-scale dry anaerobic digestion plant and from lab-scale biomethanation tests), the obtained experimental results did not show significant increase in methane yield related to the presence of biochar (330.40 NL CH4 kgVS-1 using plant inoculum; 335.41 NL CH4 kgVS-1 using plant inoculum with biochar, 311.78 NL CH4 kgVS-1 using lab-inoculum and 366.43 NL CH4 kgVS-1 using lab-inoculum with biochar), but led to significant changes in the microbial community composition. These results are likely related with the specific biochar physical-chemical features and low adsorption potential. Resulting digestate quality was also investigated: biochar-enriched digestates were characterized by increased biological stability (809 ± 264 mg O2 kgVS-1 h-1 vs. 554 ± 76 mg O2 kgVS-1 h-1 for biochar-free and biochar-enriched digestates, respectively), lower heavy metals concentrations (with the exception of Cd), but higher polycyclic aromatic hydrocarbons content, with a reported maximum concentration of 8.9 mgPAH kgTS-1 for biochar-enriched digestate derived from AD test with lab-inoculum, which could trigger non-compliance with regulation limits for agricultural reuse of digestates. However, phytotoxicity assessments showed a decreased toxicity of biochar-containing digestates when compared to biochar-free digestates.

Effects of char from biomass gasification on carbon retention and nitrogen conversion in landfill simulation bioreactors.

The application of char from biomass gasification as a filling material in landfill simulation reactors was investigated to evaluate the effect of char on carbon retention and nitrogen leaching, nitrogen denitrification, and waste stabilization. Landfill simulation columns filled with fine fraction of aged refuse (AR) and solid fraction of digestate (SFD) were used, with two char application methods: embedding a char layer between AR and SFD layers and mixing char with the SFD. The experimental results show that char application increased the biodegradable organic matter content as the respiration index (RI4) of the mixture char-SFD increased up to 37.7%, which could enhance the heterotrophic denitrification. Moreover, 12.3% of ammonia leaching was avoid by applying the SFD mixed with char. These results indicate that char from biomass gasification poses a significant enhancement on nitrogen and carbon retention which might increase the denitrification capacity of the SFD in the long run. Although high nitrogen removal rates were achieved (up to 23.1 mg N/kg-TS day), the addition of char from biomass gasification has little effect on the nitrate removal.

Statistical analysis for the quality assessment of digestates from separately collected organic fraction of municipal solid waste (OFMSW) and agro-industrial feedstock. Should input feedstock to anaerobic digestion determine the legal status of digestate?

Management options for digestate produced by anaerobic digestion plants influence the environmental and economic sustainability of the biogas sector. Further, digestate can be both used or disposed of according to its legal classification: that is, waste or by-product, or product (by using End of Waste procedure). Currently, legal digestate status is decided by EU member states on a case-by-case basis, according to specific positive lists of input feedstocks and quality requirements in terms of physical properties and chemical concentrations. Biased exclusion of input feedstock can force digestate to a specific waste classification and undergo post-treatment and disposal options that can negatively affect the profitability of biogas installations. This is the case of the Italian regulation, where the positive list of input feedstock excludes a priori separately collected organic fractions of municipal solid waste (OFMSW), while including agro-industrial residues (AGRO). This study determined the differences between the two digestate typologies (OFMSW versus AGRO) through statistical analysis, implemented on a dataset, designed to gather data about digestate's physical-chemical parameters from relevant scientific literature and unpublished private databases. The datasets consisted of 190 entries, derived from more than 2,000 samples. Further, the study provided a compliance assessment between the resulting parameter means and the current regulation limits. Upper confidence limits for the means (level of significance α = 0.05) calculated for both digestate typologies were found to be compliant with the legal requirements. Therefore, no statistical ratio seems to support the difference in the legislative approach as proposed by Italian law-makers. OFMSW resulted significantly different from AGRO for VS (650.1 g/kg TS vs. 843.8 g/kg TS, respectively), N-NH4 (81.9 g/kg TS vs. 46.19 g/kg TS), N-TOT (109.7 g/kg TS vs. 65.32 g/kg TS), P-TOT (7.22 g/kg TS vs. 21.9 g/kg TS), Pb (18.6 mg/kg TS vs. 4.66 mg/kg TS), Ni (11.03 mg/kg TS vs. 8.20 mg/kg TS), Cr-TOT (12.74 mg/kg TS vs. 8.74 mg/kg TS) and Hg (0.08 mg/kg TS vs. 0.05 mg/kg TS). However, the statistical analysis must be implemented on a wider set of parameters not covered by this study (e.g. ecotoxicological features).

Assessment of the ecotoxicity of phytotreatment substrate soil as landfill cover material for in-situ leachate management.

Phytotreatment capping in closed landfills is a promising, cost-effective, in situ option for sustainable leachate treatment and might be synergistically coupled with energy crops to produce renewable energy (e.g.: biodiesel or bioethanol). This study proposes to use 0.30 m of soil as growing substrate for plants cultivated on the temporary cover of closed landfills. Once the leachate phytotreatment process is no longer required, 0.70 m of the same soil would be added to attain the final top cover configuration. This solution would entail saving the costs of excavation and backfilling. However, worsening of the initial soil quality due to potential contaminant transfer from the liquid to the solid matrix must be avoided because EU legislation (such as that in Italy) fixes concentration limits for contaminants in soil. In this research, samples of soil used as substrate in a lab-scale leachate phytotreatment test with sunflowers were analysed to provide chemical characterization before, during, and at the end of the experiment. The results showed that the phytotreatment activity did not increase initial contaminant concentrations. These results are reinforced by those from ecotoxicological bioassays in which Eisenia fetida (earthworms), Lepidium sativum (cress), Folsomia candida (collembola), and Caenorhabditis elegans and Steinernema carpocapsae (nematodes) were used. It was observed that, by the end of the experiment, the substrate soil did not affect the earthworms, collembola and nematode behaviour, or the growth of cress.

Stabilization of solid digestate and nitrogen removal from mature leachate in landfill simulation bioreactors packed with aged refuse.

Digestate from biogas plants managing municipal solid waste needs to be stabilized prior to final utilization or disposal. Based on the concept of urban mining, aged refuse from a closed landfill was used to treat landfill leachate, but nitrogen removal by biological denitrification was limited. The aim of this study was to use a digestate layer in bioreactors containing aged refuse to enhance the biological denitrification capacity of the aged refuse, stabilize digestate, and mitigate the ammonia emissions from digestate leaching with leachate recirculation. Six identical landfill columns filled with 0% (R0), 5% (R5), and 15% (R15) of solid digestate above aged refuse (ratios based on Total Solids) were setup and nitrified leachate was periodically fed and recirculated to the columns. The nitrate removal rate in R5 and R15 was 3.4 and 10 times higher relative to the control (no digestate added). A 31.5-35.9% increase of solid digestate biostability was confirmed by tests performed under both aerobic and anaerobic conditions. The results showed that instead of land use, the solid fraction of digestate could be utilized as an inexpensive functional layer embedded in an old landfill site to enhance the denitrification capacity and achieve digestate stabilization with minimal ammonia leaching from digestate.

Dark fermentation metabolic models to study strategies for hydrogen consumers inhibition.

A Flux Balance Analysis (FBA) metabolic model of dark fermentation was developed for anaerobic mixed cultures. In particular, the model was applied to evaluate the effect of a specific inoculum pre-treatment strategy, addition of waste frying oil (WFO) on H2-producing and H2-consuming metabolic pathways. Productions of volatile fatty acid (VFAs), CO2, H2 and CH4 measured through triplicate batch experiments, were used as constraints for the FBA model, to compute fluxes trough different metabolic pathways. FBA model could estimate the effect of pre-treatment with WFO on major microbial populations present in the mixed community (H2 producing bacteria, homoacetogen and methanogens). Results revealed that low concentrations of WFO did not completely inhibited hydrogenotrophic methanogens. FBA showed that acetoclastic methanogens were more sensitive to WFO, in comparison to hydrogenotrophic methanogens. The proposed model can be used to study H2 production by any other mixed microbial culture with similar substrates.

Methane oxidation and attenuation of sulphur compounds in landfill top cover systems: Lab-scale tests.

In this study, a top cover system is investigated as a control for emissions during the aftercare of new landfills and for old landfills where biogas energy production might not be profitable. Different materials were studied as landfill cover system in lab-scale columns: mechanical-biological pretreated municipal solid waste (MBP); mechanical-biological pretreated biowaste (PB); fine (PBSf) and coarse (PBSc) mechanical-biological pretreated mixtures of biowaste and sewage sludge, and natural soil (NS). The effectiveness of these materials in removing methane and sulphur compounds from a gas stream was tested, even coupled with activated carbon membranes. Concentrations of CO2, CH4, O2, N2, H2S and mercaptans were analysed at different depths along the columns. Methane degradation was assessed using mass balance and the results were expressed in terms of methane oxidation rate (MOR). The highest maximum and mean MOR were observed for MBP (17.2gCH4/m2/hr and 10.3gCH4/m2/hr, respectively). Similar values were obtained with PB and PBSc. The lowest values of MOR were obtained for NS (6.7gCH4/m2/hr) and PBSf (3.6gCH4/m2/hr), which may be due to their low organic content and void index, respectively. Activated membranes with high load capacity did not seem to have an influence on the methane oxidation process: MBP coupled with 220g/m2 and 360g/m2 membranes gave maximum MOR of 16.5gCH4/m2/hr and 17.4gCH4/m2/hr, respectively. Activated carbon membranes proved to be very effective on H2S adsorption. Furthermore, carbonyl sulphide, ethyl mercaptan and isopropyl mercaptan seemed to be easily absorbed by the filling materials.

Energy crops on landfills: functional, environmental, and costs analysis of different landfill configurations.

The cultivation of energy crops on landfills represents an important challenge for the near future, as the possibility to use devalued sites for energy production is very attractive. In this study, four scenarios have been assessed and compared with respect to a reference case defined for northern Italy. The scenarios were defined taking into consideration current energy crops issues. In particular, the first three scenarios were based on energy maximisation, phytotreatment ability, and environmental impact, respectively. The fourth scenario was a combination of these characteristics emphasised by the previous scenarios. A multi-criteria analysis, based on economic, energetic, and environmental aspects, was performed. From the analysis, the best scenario resulted to be the fourth, with its ability to pursue several objectives simultaneously and obtain the best score relatively to both environmental and energetic criteria. On the contrary, the economic criterion emerges as weak, as all the considered scenarios showed some limits from this point of view. Important indications for future designs can be derived. The decrease of leachate production due to the presence of energy crops on the top cover, which enhances evapotranspiration, represents a favourable but critical aspect in the definition of the results.

Digestate application in landfill bioreactors to remove nitrogen of old landfill leachate.

Anaerobic digestion of organics is one of the most used solution to gain renewable energy from waste and the final product, the digestate, still rich in putrescible components and nutrients, is mainly considered for reutilization (in land use) as a bio-fertilizer or a compost after its treatment. Alternative approaches are recommended in situations where conventional digestate management practices are not suitable. Aim of this study was to develop an alternative option to use digestate to enhance nitrified leachate treatment through a digestate layer in a landfill bioreactor. Two identical landfill columns (Ra and Rb) filled with the same solid digestate were set and nitrified leachate was used as influent. Ra ceased after 75 day's operation to get solid samples and calculate the C/N mass balance while Rb was operated for 132 days. Every two or three days, effluent from the columns were discarded and the columns were refilled with nitrified leachate (average N-NO3-concentration = 1,438 mg-N/L). N-NO3- removal efficiency of 94.7% and N-NO3- removal capacity of 19.2 mg N-NO3-/gTS-digestate were achieved after 75 days operation in Ra. Prolonging the operation to 132 days in Rb, N-NO3- removal efficiency and N-NO3- removal capacity were 72.5% and 33.1 mg N-NO3-/gTS-digestate, respectively. The experimental analysis of the process suggested that 85.4% of nitrate removal could be attributed to denitrification while the contribution percentage of adsorption was 14.6%. These results suggest that those solid digestates not for agricultural or land use, could be used in landfill bioreactors to remove the nitrogen from old landfill leachate.

Pre-treating anaerobic mixed microflora with waste frying oil: A novel method to inhibit hydrogen consumption.

An innovative method was introduced to inhibit methanogenic H2 consumption during dark fermentative hydrogen production by anaerobic mixed cultures. Waste frying oil was used as an inhibitor for hydrogenotrophic methanogens. Simultaneous effect of waste frying oil concentrations (0-20 g/L) and initial pH (5.5, 6.5 and 7.5) on inhibition of methanogenic H2 consumption and enhancement of H2 accumulation were investigated using glucose as substrate. Enhanced hydrogen yields with decreased methane productions were observed with increasing the waste frying oil concentrations. On average, CH4 productions from glucose in the cultures received 10 g/L WFO were reduced by 88%. Increased WFO concentration up to 20 g/L led to negligible CH4 productions and in turn enhanced H2 yields. Hydrogen yields of 209.26, 195.35 and 185.60 mL/g glucoseadded were obtained for the cultures pre-treated with 20 g/L waste frying oil with initial pH of 5.5, 6.5 and 7.5 respectively. H2 production by pre-treated cultures was also studied using a synthetic food waste. Anaerobic mixed cultures were pre-treated with 10 g/L WFO and varying durations (0, 24 and 48 h). A H2 yield of 71.46 mL/g VS was obtained for cultures pre-treated with 10 g/L WFO for 48 h that was 475% higher than untreated control. This study suggests a novel and inexpensive approach for suppressing hydrogenotrophic methanogens during dark fermentative H2 production.