Nutrient recovery and recycling from fishery waste and by-products.

The circular bio-based economy offers great untapped potential for the food industry as possible valuable products and energy can be recovered from food waste. This can promote more sustainable and resilient food systems in Europe in follow-up of the European Commission's Farm to Fork strategy and support the global transition to more sustainable agri-food systems with the common agricultural and fisheries policies. With its high nutrient content, waste and by-products originating from fish and seafood industry (including aquaculture) are one of the most promising candidates to produce alternative fertilising products which can play a crucial role to replace synthetic mineral fertilisers. Whereas several studies highlighted the opportunities to recover valuable compounds from fishery waste, study towards their potential for the production of fertilising products is still scarce. This study presents an extensive overview of the characteristics of fishery waste and by-products (i.e., fish processing waste, fish sludge, seafood waste/by-products), the state-of-the-art nutrient recovery technologies and recovered nutrients as fertilising products from these waste streams. The European Commission has already adopted a revised Fertilising Products Regulation (EU) 2019/1009 providing opportunities for fertilising products from various bio-based origins. In frame of this opportunity, we address the quality and safety aspects of the fishery waste-derived fertilising products under these criteria and highlight possible obstacles on their way to the market in the future. Considering its high nutrient content and vast abundance, fish sludge has a great potential but should be treated/refined before being applied to soil. In addition to the parameters currently regulated, it is crucial to consider the salinity levels of such fertilising products as well as the possible presence of other micropollutants especially microplastics to warrant their safe use in agriculture. The agronomic performance of fishery waste-derived fertilisers is also compiled and reported in the last section of this review paper, which in most cases perform equally to that of conventional synthetic fertilisers.

Catchment-wide validated assessment of combined sewer overflows (CSOs) in a mediterranean coastal area and possible disinfection methods to mitigate microbial contamination.

The first phase of this study aimed to evaluate the environmental impact of combined sewer overflow (CSO) events originated from 35 spillways on the Rio Vallescura catchment (Central Italy) and to understand their contribution to the deterioration of the coastal bathing water quality. A specific analytical campaign was carried out in the sewer system and a dynamic rainfall-runoff simulation model was developed and integrated with a water quality model and further validated. The simulations led to identify the most critical spills in terms of flow rate and selected pollutant loads (i.e. suspended solids, biochemical oxygen demand, chemical oxygen demand, total Kjeldahl nitrogen, Escherichia coli). Specifically, the E. coli release in the water body due to CSO events represented almost 100% of the different pollutant sources considered. In the second phase, the applicability of various disinfection methods was investigated on the CSOs introduced into the catchment. On site physical (UV) and lab-scale chemical (peracetic acid (PAA), performic acid (PFA), ozone) disinfectant agents were tested on microbial indicators including E. coli and intestinal enterococci. PFA and ozone were more effective on the removal of both bacteria (above 3.5 log units) even at low concentration and with short contact time; whereas, PAA showed a moderate removal efficiency (around 2.5 log units) only for E. coli. The highest removal efficiency was achieved in the on-site UV unit and none of the indicator bacteria was detected in the final effluent after the sand filtration and UV treatment. Finally, potential scenarios were developed in comparison to the baseline scenario for the management and treatment of CSOs where a mitigation of E. coli loads from 28% to 73% was achieved on the receiving water body, and a comparative cost assessment of the disinfection methods was provided for in situ treatment of the most critical spillway.

Brine treatment technologies towards minimum/zero liquid discharge and resource recovery: State of the art and techno-economic assessment.

In the framework of minimum liquid discharge (MLD) or zero liquid discharge (ZLD), sustainable brine management can be achieved via appropriate hybrid treatment technologies that provide water reuse, resource recovery, energy recovery and even freshwater production. This paper reviews the state of the art brine treatment technologies targeting MLD/ZLD and resource recovery and highlights their advantages and limitations. The right combination of treatment processes can add a high value to the brine management and shift the focus from removal to recovery and reuse point and help to adopt a more circular economy approach. ZLD technologies targets 100% water recovery using both membrane- and thermal-based technologies, while they are often hindered by high cost and intensive energy requirement. Meanwhile, the recovery of salts and other resources can partially compensate the operation cost of ZLD processes. MLD is a promising option that achieves up to 95% water recovery by using mainly membrane-based technologies. At this point, feasibility assessment is important to assess the environmental and economic sound of technologies. In the second part, we provide a techno-economic assessment of the most common technologies to provide possible benefits on a desalination plant. In the latter sections, innovative brine treatment schemes are discussed aiming MLD/ZLD, while resource recovery from brine and possible valorization routes of the recovered materials are highlighted to help to reduce the overall costs of the plants and to reach the targets of circular economy.

In search of the optimal inoculum to substrate ratio during anaerobic co-digestion of spent coffee grounds and cow manure.

The inoculum to substrate (I:S) ratio is a crucial operating parameter during the start-up period of anaerobic digestion (AD) processes and this ratio shows high differentiation with respect to substrate composition. While spent coffee grounds (SCG) have started to gain attraction in AD as a co-substrate due to their vast production and promising methane potential, there is still not enough information on the operative environment of SCG-based biogas reactors. This study investigated the optimal I:S ratio during anaerobic co-digestion of SCG and cow manure. Biochemical methane potential tests were conducted at mesophilic conditions and the influence of I:S ratio on methane production and digestion stability was evaluated at a wide range of I:S ratios from 0.5:1 to 4:1 (volatile solids (VS) basis). Methane yields increased gradually starting from the I:S ratio of 0.5:1 up to 3:1 and the highest methane yield (225 mlCH4 gVS-1) was achieved at the I:S ratio of 3:1. Comparatively lower methane yields were obtained at the ratios of 3.5:1 and 4:1. Instable AD conditions were established at the lowest I:S ratio examined (0.5:1), which caused volatile fatty acid (VFA) accumulation. The results highlighted that anaerobic co-digestion of SCG and cow manure is a promising approach, while the I:S ratio should be well-maintained due to the high potential risk of rapid and/or excess VFA production of these feedstocks.

Operating conditions influence microbial community structures, elimination of the antibiotic resistance genes and metabolites during anaerobic digestion of cow manure in the presence of oxytetracycline.

The way that antibiotic residues in manure follow is one of the greatest concerns due to its potential negative impacts on microbial communities, the release of metabolites and antibiotic resistant genes (ARGs) into the nature and the loss of energy recovery in anaerobic digestion (AD) systems. This study evaluated the link between different operating conditions, the biodegradation of oxytetracycline (OTC) and the formation of its metabolites and ARGs in anaerobic digesters treating cow manure. Microbial communities and ARGs were determined through the use of quantitative real-time PCR. The biodegradation of OTC and occurrence of metabolites were determined using UV-HPLC and LC/MS/MS respectively. The maximum quantity of resistance genes was also examined at the beginning of AD tests and concentration was in the order of: tetM >tetO. The numbers of ARGs were always higher at high volatile solids (VS) content and high mixing rate. The results of the investigation revealed that relationship between mixing rate and VS content plays a crucial role for elimination of ARGs, OTC and metabolites. This can be attributed to high abundance of microorganisms due to high VS content and their increased contact with elevated mixing rate. An increased interaction between microorganisms triggers the promotion of ARGs.

Application of next-generation sequencing methods for microbial monitoring of anaerobic digestion of lignocellulosic biomass.

The anaerobic digestion of lignocellulosic wastes is considered an efficient method for managing the world's energy shortages and resolving contemporary environmental problems. However, the recalcitrance of lignocellulosic biomass represents a barrier to maximizing biogas production. The purpose of this review is to examine the extent to which sequencing methods can be employed to monitor such biofuel conversion processes. From a microbial perspective, we present a detailed insight into anaerobic digesters that utilize lignocellulosic biomass and discuss some benefits and disadvantages associated with the microbial sequencing techniques that are typically applied. We further evaluate the extent to which a hybrid approach incorporating a variation of existing methods can be utilized to develop a more in-depth understanding of microbial communities. It is hoped that this deeper knowledge will enhance the reliability and extent of research findings with the end objective of improving the stability of anaerobic digesters that manage lignocellulosic biomass.

Changes in microbial community structures due to varying operational conditions in the anaerobic digestion of oxytetracycline-medicated cow manure.

Management of manure containing veterinary antibiotics is a major concern in anaerobic treatment systems because of their possible adverse effects on microbial communities. Therefore, the aim of study was to investigate how oxytetracycline (OTC) influences bacteria and acetoclastic and hydrogenotrophic methanogens under varying operational conditions in OTC-medicated and non-medicated anaerobic cow manure digesters. Concentrations of OTC and its metabolites throughout the anaerobic digestion were determined using ultraviolet-high-performance liquid chromatography (UV-HPLC) and tandem liquid chromatography-mass spectrometry (LC/MS/MS), respectively. Fluorescent in situ hybridization, denaturing gradient gel electrophoresis, cloning, and sequencing analyses were used to monitor changes in microbial community structures. According to the results of analytical and molecular approaches, operating conditions highly influence active microbial community dynamics and associate with biogas production and elimination of OTC and its metabolites during anaerobic digestion of cow manure in the presence of an average initial concentration of 2.2 mg OTC/L. The impact of operating conditions has a drastic effect on acetoclastic methanogens than hydrogenotrophic methanogens and bacteria.

Degradation of oxytetracycline and its impacts on biogas-producing microbial community structure.

The effect of veterinary antibiotics in anaerobic digesters is a concern where methane production efficiency is highly dependent on microbial community structure. In this study, both anaerobic degradation of a common veterinary antibiotic, oxytetracycline (OTC), and its effects on an anaerobic digester microbial community were investigated. Qualitative and quantitative molecular tools were used to monitor changes in microbial community structure during a 60-day batch incubation period of cow manure with the addition of different concentrations of the antibiotic. Molecular data were interpreted by a further redundancy analysis as a multivariate statistics approach. At the end of the experiment, approximately 48, 33, and 17 % of the initially added 50, 100, and 200 mg l(-1) of OTC was still present in the serum bottles which reduced the biogas production via accumulation of some of the volatile fatty acids (VFAs). Biogas production was highly correlated with Methanobacteriales and Methanosarcinales gene copy numbers, and those parameters were negatively affected with oxytetracycline and VFA concentrations.

Acidification of non-medicated and oxytetracycline-medicated cattle manures during anaerobic digestion.

Possible adverse effects of a commonly used veterinary antibiotic, oxytetracycline (OTC), on acidogenic phase of anaerobic digestion of cattle manure along with optimum operating conditions were investigated. A standard veterinary practice of 50 ml OTC solution (20 mg/kg cattle weight) was injected into the muscles of cattle and then manure samples were collected for 5 days following the injection. The 5-day samples were equally mixed and used throughout digestion experiments. Preliminary batch tests were conducted to obtain the optimum pH range and observe volatile fatty acids (VFAs) production. In this regard, different sets of batch digesters were operated at pH ranging from 5.2 +/- 0.1 to 5.8 +/- 0.1 at mesophilic conditions with total solids content of 6.0 +/- 0.2%. The pH of 5.5 +/- 0.1 was found to be the optimum value for acidification for both non-medicated and OTC-medicated conditions. Under predetermined conditions, maximum total VFA (VFAtot) of 830 +/- 3 mg (as acetic acid)/L was produced and maximum acidification rate was evaluated as 11% for OTC-medicated cattle manure, whereas they were 900 +/- 6 mg (as acetic acid)/L and 12% for non-medicated manure. Digestion studies were further continued in a semi-continuous mode at pH 5.5 +/- 0.1 and SRT/HRT of 5 days. VFAtot concentrations and maximum acidification rate increased up to 2181 +/- 19 mg (as acetic acid)/L and 29% for non-medicated cattle manure. For OTC-medicated cattle manure, lower acidification rate of 18% was observed.

Post-treatment of high-rate activated sludge effluent via zeolite adsorption and recovery of ammonium-nitrogen as alternative fertilising products.

This study investigates the potential to connect nutrient flows between wastewater treatment and agriculture through a two-stage nitrogen (N) recovery system composed of high-rate activated sludge treatment in contact stabilisation mode (HRAS/CS) and column adsorption with zeolite. The HRAS/CS process removes organic matter and suspended solids in wastewater, leaving N behind in the effluent. The N was successfully recovered with the zeolite column under different scenarios, generating N and K-rich by-products. The regeneration effluent from the zeolite column with KCl contained 60-845 mg NH4+-N/L and 1.6-14.3 g K/L, having potential for use as fertigation water. The N-saturated zeolite contained 1.5-8.4 mg N/g and 14.3-19.3 mg K/g of the product fresh weight and low contaminant content, making it potentially eligible as various fertilising products. Adsorption can thus concentrate N from HRAS/CS effluent and produce by-products with potential agricultural value while meeting chemical oxygen demand and total nitrogen discharge standards.

Potential availability of metals in anaerobic mono- and co-digestion of pig manure and maize.

This study aims to analyse the potential availability of essential metals including as Co, Fe, Ni, Zn, Mn, and Cu and non-essential metals such as Pb, Cr, and Cd within anaerobic mono- and co-digestion of pig manure and maize. The metals partitioning was determined using the Modified BCR (European Community Bureau of Reference) sequential extraction at defined intervals over a 45-days period, correlating changes in metals speciation with key digestion variables. The findings revealed that Cr, Cu, Fe, and Pb were predominantly associated with the oxidisable fraction, while Zn, Mn, and Cd were potentially available in both processes. Notably, NH4+-N and the VFAs, except propionic acid, correlated significantly with the available fractions of Co, Mn, Ni, Zn, Cr, and Pb during mono-digestion of pig manure. The wider pH range and the chemical properties of the feedstock in co-digestion resulted in varied correlations between the metals availability and the digestion variables.

From grass to gas and beyond: Anaerobic digestion as a key enabling technology for a residual grass biorefinery.

Roadside grass clippings hold potential as a sustainable source of bioenergy as they do not compete with crops for land use, and are only partially utilized for low-value applications. In this study, we proposed using roadside grass as a sole feedstock for anaerobic digestion (AD) in three different settings, and assessed the potential of producing biomaterials and fertilizers from grass-based digestate. Wet continuous digestion at pilot scale and dry batch digestion at pilot and large scales resulted in biogas yields up to 700 Nm3.t-1 DOM with a methane content of 49-55 %. Despite promising results, wet AD had operational problems such as clogging and poor mixing; once upscaled, the dry digestion initially also presented an operational problem with acidification, which was overcome by the second trial. Digested grass fibers from the pilot dry AD were processed into biomaterials and performed similarly or better than the undigested fibers, while around 20 % performance reduction was observed when compared to reference wood fibers. A mass balance indicated reduced fiber recovery when higher biogas production was obtained. The liquid fraction from the pilot dry AD was characterized for its nutrient content and used as a biofertilizer in another study. In contrast, the leachate collected from the large-scale dry AD had a low nitrogen content and high chloride content that could hinder its further use. Finally, a regional market analysis was conducted showing that the biocomposites produced with the available grass fibers could substitute at least half of the current European market based on our results.

Monitoring of cyanobacterial blooms and assessing polymer-enhanced microfiltration and ultrafiltration for microcystin removal in an Italian drinking water treatment plant.

The water intake of a drinking water treatment plant (DWTP) in Central Italy was monitored over six bloom seasons for cyanotoxin severity, which supplies drinking water from an oligo-mesotrophic lake with microcystin levels up to 10.3 µg/L. The historical data showed that the water temperature did not show extreme/large seasonal variation and it was not correlated either with cyanobacterial growth or microcystin concentration. Among all parameters, the cyanobacteria growth was negatively correlated with humidity and manganese and positively correlated with atmospheric temperature. No significant correlation was found between microcystin concentration and the climatic parameters. Polymer(chitosan)-enhanced microfiltration (PEMF) and ultrafiltration (PEUF) were further tested as an alternative microcystin removal approach from dense cyanobacteria-rich flows. The dominant cyanobacteria in the water intake, Planktothrix rubescens, was isolated and enriched to simulate cyanobacterial blooms in the lake. The PEMF and PEUF were separately applied to enriched P. rubescens culture (PC) (microcystin = 1.236 µg/L) as well as to the sand filter backwash water (SFBW) of the DWTP where microcystin concentration was higher than 12 µg/L. The overall microcystin removal rates from the final effluent of PC (always <0.15 µg/L) were between 90.1-94.7% and 89.5-95.4% using 4 and 20 mg chitosan/L, respectively. Meanwhile, after the PEMF and PEUF of SFBW, the final effluent contained only 0.099 and 0.057 µg microcystin/L with an overall removal >99%. The presented results are the first from the application of chitosan to remove P. rubescens as well as the implementation of PEMF and PEUF on SFBW to remove cyanobacterial cells and associated toxins.

Microplastics in real wastewater treatment schemes: Comparative assessment and relevant inhibition effects on anaerobic processes.

The occurrence, fate and removal of microplastics (MPs) in a wastewater treatment plant (WWTP) in Central Italy were investigated together with their potential adverse effects on anaerobic processes. In the influent of the WWTP, 3.6 MPs.L-1 were detected that mostly comprised polyester fibers and particles in the shape of films, ranging 0.1-0.5 mm and made of polyethylene and polypropylene (PP). The full-scale conventional activated sludge scheme removed 86% of MPs, with the main reduction in the primary and secondary settling. MPs particles bigger than 1 mm were not detected in the final effluent and some loss of polymers types were observed. In comparison, the pilot-scale upflow granular anaerobic sludge blanket (UASB) + anaerobic membrane bioreactor (AnMBR) configuration achieved 94% MPs removal with the abatement of 87% of fibers and 100% of particles. The results highlighted an accumulation phenomenon of MPs in the sludge and suggested the need to further investigate the effects of MPs on anaerobic processes. Accordingly, PP-MPs at concentrations from 5 PP-MPs.gTS-1 to 50 PP-MPs.gTS-1 were spiked in the pilot-scale UASB reactor that was fed with real municipal wastewater, where up to 58% decrease in methanogenic activity was observed at the exposure of 50 PP-MPs.gTS-1. To the best of our knowledge, the presented results are the first to report of PP-MPs inhibition on anaerobic processes.

Crop-based composting of lignocellulosic digestates: Focus on bacterial and fungal diversity.

In this study, organic matter degradation and microbial diversity were assessed during the composting of lignocellulose-rich digestates. Digestates were collected based on each crop type during anaerobic co-digestion of cow manure and barley, triticale, wheat and rye. Bacterial and fungal diversity in digestate composting systems were determined by 16S and 18S rRNA gene amplicon sequencing, respectively. Crop-based composting of anaerobic digestates showed similar process trends in terms of pH, temperature, moisture content (MC) and C:N ratio. The properties of final compost products were in accordance with the national legislations regarding soil applications, except MC, which were therefore air-dried before being amended to soil. Most abundant bacterial genera were represented by Luteimonas, Bacillus, Ochrobactrum and Thermobifida. Meanwhile, Thermomyces, Aspergillus, Galactomyces and Neurospora were detected as the predominant fungal genera in all compost samples.

Linking nano-ZnO contamination to microbial community profiling in sanitary landfill simulations.

Nanomaterials (NMs) commercially used for various activities mostly end up in landfills. Reduced biogas productions reported in landfill reactors create a need for more comprehensive research on these greatly-diverse microbial pools. In order to evaluate the impact of one of the most widely-used NMs, namely nano-zinc oxide (nano-ZnO), simulated bioreactor and conventional landfills were operated using real municipal solid waste (MSW) for 300 days with addition nano-ZnO. Leachate samples were taken at different phases and analyzed by 16S rRNA gene amplicon sequencing. The bacterial communities were distinctly characterized by Cloacamonaceae (phylum WWE1), Rhodocyclaceae (phylum Proteobacteria), Porphyromonadaceae (phylum Bacteroidetes), and Synergistaceae (phylum Synergistetes). The bacterial community in the bioreactors shifted at the end of the operation and was dominated by Rhodocyclaceae. There was not a major change in the bacterial community in the conventional reactors. The methanogenic archaeal diversity highly differed between the bioreactors and conventional reactors. The dominance of Methanomicrobiaceae was observed in the bioreactors during the peak methane-production period; however, their prominence shifted to WSA2 in the nano-ZnO-added bioreactor and to Methanocorpusculaceae in the control bioreactor towards the end. Methanocorpusculaceae was the most abundant family in both conventional control and nano-ZnO-containing reactors.

Fungal bioaugmentation of anaerobic digesters fed with lignocellulosic biomass: What to expect from anaerobic fungus Orpinomyces sp.

Energy-efficient biogas reactors are often designed and operated mimicking natural microbial ecosystems such as the digestive tracts of ruminants. Anaerobic fungi play a crucial role in the degradation of lignocellulose-rich fiber thanks to their high cellulolytic activity. Fungal bioaugmentation is therefore at the heart of our understanding of enhancing anaerobic digestion (AD). The efficiency of bioaugmentation with anaerobic fungus Orpinomyces sp. was evaluated in lignocellulose-based AD configurations. Fungal bioaugmentation increased the methane yield by 15-33% during anaerobic co-digestion of cow manure and selected cereal crops/straws. Harvesting stage of the crops was a decisive parameter to influence methane production together with fungal bioaugmentation. A more efficient fermentation process in the bioaugmented digesters was distinguished by relatively-higher abundance of Synergistetes, which was mainly represented by the genus Anaerobaculum. On the contrary, the composition of the methanogenic archaea did not change, and the majority of methanogens was assigned to Methanosarcina.

Pilot scale cellulose recovery from sewage sludge and reuse in building and construction material.

The recovery of cellulose in toilet paper from municipal wastewater is one of the most innovative actions in the circular economy context. In fact, fibres could address possible new uses in the building sector as reinforcing components in binder-based materials. In this paper, rotating belt filters were tested to enhance the recovery of sludge rich in cellulose fibres for possible valorisation in construction applications. Recovered cellulosic material reached value up to 26.6 gm-3 with maximum solids removal of 74%. Content of cellulose, hemicellulose and lignin was found averagely equal to 87% of the total composition. Predictive equation of cellulosic material was further obtained. The addition of recovered cellulose fibres in mortars bring benefits in terms of lightness, microstructure and moisture buffering value (0.17 g/m2%UR). Concerning mechanical properties, flexural strength was improved with the addition of 20% of recovered cellulose fibres. In addition, a simplified economic assessment was reported for two possible pre-mixed blends with 5% and 20% of recovered fibres content.

Bioaugmentation with Clostridium thermocellum to enhance the anaerobic biodegradation of lignocellulosic agricultural residues.

This study aimed to improve biomethane production from lignocellulosic biomass by assessing the impact of bioaugmentation with Clostridium thermocellum on the performance of anaerobic digesters at different inoculation ratios. The outputs of the digestion experiments revealed that bioaugmentation strategies with C. thermocellum increased the methane yield up to 39%. The sequencing analysis indicated that the indigenous microbial community was modified by the bioaugmentation. During the process of bioaugmentation, in the digester that was inoculated at the ratio of 20% (v:v), an increase in the abundance of Ruminococcaceae family led to a decrease in the Bacteroidaceae and Synergistaceae families. Furthermore, the metabolic products of the bioaugmented strains greatly influenced the diversity of the archaeal community and an increase in the abundance of Methanomicrobiales was observed.

Microbial monitoring of ammonia removal in a UASB reactor treating pre-digested chicken manure with anaerobic granular inoculum.

Performance and microbial community dynamics in an upflow anaerobic sludge bed (UASB) reactor coupled with anaerobic ammonium oxidizing (Anammox) treating diluted chicken manure digestate (Total ammonia nitrogen; TAN=123±10mg/L) were investigated for a 120-d operating period in the presence of anaerobic granular inoculum. Maximum TAN removal efficiency reached to above 80% with as low as 20mg/L TAN concentrations in the effluent. Moreover, total COD (tCOD) with 807±215mg/L in the influent was removed by 60-80%. High-throughput sequencing revealed that Proteobacteria, Actinobacteria, and Firmicutes were dominant phyla followed by Euryarchaeota and Bacteroidetes. The relative abundance of Planctomycetes significantly increased from 4% to 8-9% during the late days of the operation with decreased tCOD concentration, which indicated a more optimum condition to favor ammonia removal through anammox route. There was also significant association between the hzsA gene and ammonia removal in the UASB reactor.