Growth of microalgae and cyanobacteria consortium in a photobioreactor treating liquid anaerobic digestate from vegetable waste.

This research examines the biological treatment of undiluted vegetable waste digestate conducted in a bubble column photobioreactor. Initially, the bioreactor containing 3N-BBM medium was inoculated with Microglena sp., Tetradesmus obliquus, and Desmodesmus subspicatus mixture with a density of 1.0 × 104 cells/mL and the consortium was cultivated for 30 days. Then, the bioreactor was semi-continuously fed with liquid digestate with hydraulic retention time (HRT) of 30 days, and the treatment process was continued for the next 15 weeks. The change in the microalgal and cyanobacterial species domination was measured in regular intervals using cell counting with droplet method on a microscope slide. At the end of the experiment, Desmonostoc sp. cyanobacteria (identified with 16S ribosomal RNA genetical analysis) as well as Tetradesmus obliquus green algae along with Rhodanobacteraceae and Planococcaceae bacteria (determined with V3-V4 16sRNA metagenomic studies) dominated the microbial community in the photobioreactor. The experiment demonstrated high treatment efficiency, since nitrogen and soluble COD were removed by 89.3 ± 0.5% and 91.2 ± 1.6%, respectively, whereas for phosphates, 72.8 ± 2.1% removal rate was achieved.

Effect of solar and artificial lighting on microalgae cultivation and treatment of liquid digestate.

A comparative study was carried out to assess the effect of two light sources on microalgae cultivation and the treatment of liquid digestate. The R1 photobioreactor operated with LED lightning allowed to achieve moderate nutrient removal rates whereas soluble COD (Chemical Oxygen Demand) was reduced in 90%. After switching this reactor into sunlight, the removal rate of phosphates increased to 66%. However, the greatest removal rate of both nutrients and sCOD of up to 93% was observed in the R2 photobioreactor operated only under sunlight. Microglena sp. was the dominant algae growing in the R1 reactor, and the main bacteria families detected were Chitinophagaceae, Sphingomonadaceae and Xanthobacteraceae. In contrast, Tetradesmus obliquus dominated in the R2 reactor and Rhodanobacteraceae, Chitinophagaceae and A4b were predominant bacteria in this run. Furthermore, much greater biomass productivity as well as overall biomass density was observed in the R2 photobioreactor cultivated exclusively with solar lightning.

Anaerobic co-digestion of kitchen waste with hyperthermophilically pretreated grass for biohydrogen and biomethane production.

Anaerobic digestion of kitchen waste with grass after hyperthermophilic pretreatment was performed in semi-continuously operated reactors. The greatest methane yield of 293 NmlCH4/gVS (volatile solids) was reported for the mixture of both substrates at 55 °C with a solids retention time of 30 d and the corresponding organic lading rate of 1.72 kgVS/m3/d. In contrast, pretreated grass subjected to thermophilic digestion produced only 131 NmlCH4/gVS. However, when mesophilic conditions were applied, the digestion process turned into dark fermentation, especially visible for the mixture. Metagenomic analysis revealed the dominance Ruminococcaceae, Atopobiaceae and Lactobacillaceae at a family level in mesophilic processes, whereas Petrotogaceae, Synergistaceae, Hungateiclostridiaceae, Planococcaceae and two methanogens Methanosarcinaceae and Methanothermobacteriaceae were the most frequent microbes of thermophilic digestion. Kitchen waste can successfully be co-digested with hyperthermophilically pretreated grass at high loading rates, however the digesters must be operated at thermophilic temperatures.

Treatment of Liquid Digestate by Green Algal Isolates from Artificial Eutrophic Pond.

The ability of aquatic microalgae to treat the liquid digestate obtained from the anaerobic digestion of plant waste was investigated. Microalgae were isolated from natural environment for a laboratory-scale cultivation and were then used to remove nutrients and organic contaminants from the liquid digestate. It was shown that the microalgae consortia (Tetradesmus obliquus, Microglena sp., Desmodesmus subspicatus) could reduce nitrogen, phosphates, and total COD by up to 70%, 57%, and 95%, respectively. A new algae genus Microglena was isolated, which in a consortium with Tetradesmus obliquus and Desmodesmus subspicatus exhibited a high efficiency in the removal of both organic contaminants and nutrients from the liquid fraction of digestate.

Isolation and Use of Coprothermobacter spp. to Improve Anaerobic Thermophilic Digestion of Grass.

The isolation of microorganisms was performed from digestate from the process of the anaerobic digestion (AD) of grass after hyperthermophilic pretreatment. The bacterium that was isolated and identified was Coprothermobacter sp. Using the isolated bacteria, an AD process on fresh grass (GB) and pretreated grass (PGB) was carried out with 10% of its addition. The highest methane yield of 219 NmlCH4/gVS was recorded for PGB at 55 °C. In contrast, fresh grass subjected to thermophilic digestion produced only 63 NmlCH4/gVS. Due to the addition of bacteria in the AD process, an increase in the efficiency of hydrogen and methane production was observed in both fresh grass and grass after pretreatment.

Volatile Organic Compounds and Physiological Parameters as Markers of Potato (Solanum tuberosum L.) Infection with Phytopathogens.

The feasibility of early disease detection in potato seeds storage monitoring of volatile organic compounds (VOCs) and plant physiological markers was evaluated using 10 fungal and bacterial pathogens of potato in laboratory-scale experiments. Data analysis of HS-SPME-GC-MS revealed 130 compounds released from infected potatoes, including sesquiterpenes, dimethyl disulfide, 1,2,4-trimethylbenzene, 2,6,11-trimethyldodecane, benzothiazole, 3-octanol, and 2-butanol, which may have been associated with the activity of Fusarium sambucinum, Alternaria tenuissima and Pectobacterium carotovorum. In turn, acetic acid was detected in all infected samples. The criteria of selection for volatiles for possible use as incipient disease indicators were discussed in terms of potato physiology. The established physiological markers proved to demonstrate a negative effect of phytopathogens infecting seed potatoes not only on the kinetics of stem and root growth and the development of the entire root system, but also on gas exchange, chlorophyll content in leaves, and yield. The negative effect of phytopathogens on plant growth was dependent on the time of planting after infection. The research also showed different usefulness of VOCs and physiological markers as the indicators of the toxic effect of inoculated phytopathogens at different stages of plant development and their individual organs.

Enzymatic Pretreatment of Byproducts from Soapstock Splitting and Glycerol Processing for Improvement of Biogas Production.

This study investigated acid splitting wastewater (ASW) and interphase (IF) from soapstock splitting, as well as matter organic non glycerol (MONG) from glycerol processing, as potential substrates for biogas production. Batch and semicontinuous thermophilic anaerobic digestion experiments were conducted, and the substrates were preliminary treated using commercial enzymes kindly delivered by Novozymes A/C. The greatest enhancement in the batch digestion efficiency was achieved when three preparations; EversaTransform, NovoShape, and Lecitase were applied in the hydrolysis stage, which resulted in the maximum methane yields of 937 NL/kg VS and 915 NL/kg VS obtained from IF and MONG, respectively. The co-digestion of 68% ASW, 16% IF, and 16% MONG (wet weight basis) performed at an organic loading rate (OLR) of 1.5 kg VS/m3/day provided an average methane yield of 515 NLCH4/kg VSadded and a volatile solid reduction of nearly 95%. A relatively high concentration of sulfates in the feed did not significantly affect the digestion performance but resulted in an increased hydrogen sulfide concentration in the biogas with the peak of 4000 ppm.

Dark fermentative hydrogen production from hydrolyzed sugar beet pulp improved by nitrogen and phosphorus supplementation.

The effect of nitrogen and phosphorous addition on hydrogen production from hydrolyzed Sugar beet pulp (SBP) was investigated using (NH4)3PO4, NH4Cl and K3PO4 as the supplements. In batch tests, the maximal hydrogen production of 279 dm3/kgVS was observed for K3PO4, which was added to SBP in a dose of 1 g/dm3. In semi-continuous experiments, the greatest hydrogen production of 36 dm3/kgVS was reported for the same supplement, and this value was twice higher than that of the control run. The analysis of microbiota revealed that the majority of bacteria was affiliated to the orders Clostridiales, Lactobacillales and Coriobacteriales. Moreover, a noticeable methane production was associated with the activity of Methanosphaera sp., which could grow in a low pH environment of dark fermentation.

Effect of hyperthermophilic pretreatment on methane and hydrogen production from garden waste under mesophilic and thermophilic conditions.

Anaerobic digestion of garden waste was investigated in a two-stage process consisting of hyperthermophilic pretreatment followed by mesophilic or thermophilic fermentation. The greatest digestion performance was achieved when the substrates were first treated at 70 °C for 3 days with no inoculation, and then mixed with inoculum (anaerobic sludge) and subjected to anaerobic digestion at 55 °C. Under such conditions, the maximum methane and hydrogen yields from grass were 517 NmlCH4/kgVS and 52 NmlH2/kgVS, whereas the corresponding values for leaves were 421 NmlCH4/kgVS and 23 NmlH2/kgVS, and these figure were far greater than the yields obtained in experiments with no hyperthermophilic stage. A metagenomic analysis of hyperthermophilic environments revealed the appearance of thermophilic and hyperthermophilic bacteria showing hydrolytic activity against lignocellulosic materials, including Caldicellulosiruptor, Thermovenabulum, Thermoanaerobacter, Moorella, Tepimicrobium, Geobacillus and Thermobacillus at a genus level. A noticeable methane production in the hyperthermophilic stage could be linked to the presence of Methanothermobacter sp.

Dark fermentative hydrogen production from hydrolyzed sugar beet pulp improved by iron addition.

This study evaluated the impact of three different iron compounds (Fe2O3, FeSO4, FeCl3) on hydrogen production via mesophilic dark fermentation (DF) of hydrolyzed sugar beet pulp (SBP). In batch tests, the maximum hydrogen yield of over 200 dm3H2/kgVS was achieved with the addition of 0.1 gFe2O3/dm3, which was twice greater than the control. In semi-continuous experiments, the highest hydrogen production of 52.11 dm3H2/kgVS combined with 19.4 dm3CH4/kgVS methane yield was obtained at a dose of 1 gFe2O3/dm3. Acetic, lactic and caproic acids were the main metabolic products of DF. Microbiological studies showed some balance between hydrogen producing microorganisms from the order Clostridiales and lactic acid producers (LAB) affiliated with the orders Lactobacillales and Coriobacteriales. Moreover, the presence of methanogens affiliated to the genera Methanobrevibacter and Methanosphaera was also documented. An interesting finding was the appearance of rare bacteria from the genus Caproiciproducens, which was responsible for increased caproic acid production.

The use of sugar beet pulp stillage for co-digestion with sewage sludge and poultry manure.

The anaerobic mesophilic co-digestion of sugar beet pulp stillage with poultry manure and municipal sewage sludge was investigated in this study. The sugar beet pulp stillage (SBPS) mono-digestion failed owing to an accumulation of volatile fatty acids, leading to a pH value lower than 5.5. A 20% addition of poultry manure to stillage allowed for stable digestion performance despite high volatile fatty acid (total volatile fatty acids) concentrations of 5500-8500 g m-3 with propionic acid being the predominant one and constituting 72%-76% total volatile fatty acids. For this mixture, the maximum methane production of 418 dm3 kgVSfed-1 was achieved when the reactor was operated at a solids retention time of 20 days and an organic loading rate of 4.25 kgVS m-3 d-1. The co-digestion of stillage with 60% municipal sewage sludge gave the average methane yield of around 357 dm3 kgVSfed-1 for all operational conditions applied, however, the methane percentage of biogas (up to 70%) was far greater than the corresponding values obtained for sugar beet pulp stillage-poultry manure co-digestion. Neither ammonia nor volatile fatty acids destabilised the biogas production, and the volatile fatty acid profile showed the dominance of acetic acid (72%-82% total volatile fatty acids) followed by propionic and butyric acids.

Enzymatic Conversion of Sugar Beet Pulp: A Comparison of Simultaneous Saccharification and Fermentation and Separate Hydrolysis and Fermentation for Lactic Acid Production.

This study compares the efficiency of lactic acid production by separate hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF) of sugar beet pulp, a byproduct of industrial sugar production. In experiments, sugar beet pulp was hydrolyzed using five commercial enzymes. A series of shake flask fermentations were conducted using five selected strains of lactic acid bacteria (LAB). The differences in the activities of the enzymes for degrading the principal sugar beet pulp components were reflected in the different yields of total reducing sugars. The highest yields after hydrolysis and the lowest quantities of insoluble residues were obtained using a mixture (1:1) of Viscozyme® and Ultraflo® Max. In the SHF process, only a portion of the soluble sugars released by the enzymes from the sugar beet pulp was assimilated by the LAB strains. In SSF, low enzyme loads led to reduction in the efficiency of sugar accumulation. The risk of carbon catabolic repression was reduced. Our results suggest that SSF has advantages over SHF, including lower processing costs and higher productivity. Lactic acid yield in SSF mode (approx. 30 g/L) was 80-90% higher than that in SHF.

Food waste co-digestion with slaughterhouse waste and sewage sludge: Digestate conditioning and supernatant quality.

In this study, the anaerobic mesophilic co-digestion of food waste (FW) with municipal sewage sludge (MSS) and slaughterhouse waste (SHW) was undertaken in 3-dm3 laboratory reactors as well as in 50-dm3 reactors operated in semi-continuous conditions. The highest methane yield of around 0.63 m3 CH4/kgVSfed was achieved for the mixture of FW and SHW treated in the laboratory digester operated at solids retention time (SRT) of 30 days, whereas the co-digestion of FW with MSS under similar operating conditions produced 0.46 m3 of methane from 1 kgVSfed. No significant differences between methane yields from laboratory digesters and large-scale reactors were reported. The conditioning tests with the digestates from reactor experiments revealed the highest efficiency of inorganic coagulants among all investigated chemicals, which applied in a dose of 10 g/kg allowed to reduce capiliary suction time (CST) of the digestate below 20 s. The combined conditioning with coagulants and bentonite did not further reduce the CST value but improved the quality of the digestate supernatant. In particular, the concentrations of suspended solids, COD as well as metals in the supernatant were considerably lowered.

Odorous Compounds from Poultry Manure Induce DNA Damage, Nuclear Changes, and Decrease Cell Membrane Integrity in Chicken Liver Hepatocellular Carcinoma Cells.

Animal breeding and management of organic wastes pose a serious problem to the health of livestock and workers, as well as the nearby residents. The aim of the present study was to determine the mechanisms of toxicity of selected common odorous compounds from poultry manure, including ammonia, dimethylamine (DMA), trimethylamine (TMA), butyric acid, phenol, and indole. We measured their genotoxic and cytotoxic activity in the model chicken cell line (LMH), in vitro, by comet assay and lactate dehydrogenase assay, respectively. We also made microscopic observations of any morphological changes in these cells by DAPI staining. Four compounds, namely ammonia, DMA, TMA, and butyric acid increased DNA damage in a dose-dependent manner (p < 0.05), reaching genotoxicity as high as 73.2 ± 1.9%. Phenol and indole induced extensive DNA damage independent of the concentration used. Ammonia, DMA, and TMA caused a dose-dependent release of lactate dehydrogenase (p < 0.05). The IC50 values were 0.02%, 0.05%, and 0.1% for DMA, ammonia and TMA, respectively. These compounds also induced nuclear morphological changes, such as chromatin condensation, shrinkage, nuclear fragmentation (apoptotic bodies), and chromatin lysis. Our study exhibited the damaging effects of odorous compounds in chick LMH cell line.

Simultaneous Saccharification and Fermentation of Sugar Beet Pulp with Mixed Bacterial Cultures for Lactic Acid and Propylene Glycol Production.

Research into fermentative production of lactic acid from agricultural by-products has recently concentrated on the direct conversion of biomass, whereby pure sugars are replaced with inexpensive feedstock in the process of lactic acid production. In our studies, for the first time, the source of carbon used is sugar beet pulp, generated as a by-product of industrial sugar production. In this paper, we focus on the simultaneous saccharification of lignocellulosic biomass and fermentation of lactic acid, using mixed cultures with complementary assimilation profiles. Lactic acid is one of the primary platform chemicals, and can be used to synthesize a wide variety of useful products, including green propylene glycol. A series of controlled batch fermentations was conducted under various conditions, including pretreatment with enzymatic hydrolysis. Inoculation was performed in two sequential stages, to avoid carbon catabolite repression. Biologically-synthesized lactic acid was catalytically reduced to propylene glycol over 5% Ru/C. The highest lactic acid yield was obtained with mixed cultures. The yield of propylene glycol from the biological lactic acid was similar to that obtained with a water solution of pure lactic acid. Our results show that simultaneous saccharification and fermentation enables generation of lactic acid, suitable for further chemical transformations, from agricultural residues.

Cytotoxicity of Odorous Compounds from Poultry Manure.

Long-term exposure and inhalation of odorous compounds from poultry manure can be harmful to farm workers and the surrounding residents as well as animals. The aim of the present study was to determine the cytotoxicity and IC50 values of common odorous compounds such as ammonium, dimethylamine, trimethylamine, butyric acid, phenol, and indole in the chick liver hepatocellular carcinoma cell line LMH (Leghorn Male Hepatoma), in vitro, using MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) and PrestoBlue cytotoxicity assays. The cells were microscopically examined for any morphological changes post treatment. Dimethylamine exhibited the strongest cytotoxic effect on LMH cells with an IC50 value of 0.06% and 0.04% after an exposure of 24 h and 48 h, respectively. Both ammonium and trimethylamine had comparable cytotoxicity and their IC50 values were 0.08% and 0.04% after 24 h and 48 h, respectively. Of note, indole had the lowest cytotoxicity as the majority of cells were viable even after 72 h exposure. Thus, the IC50 for indole was not calculated. Results achieved from both MTT and PrestoBlue assays were comparable. Moreover, the morphological changes induced by the tested odours in LMH cells resulted in monolayer destruction, cytoplasm vacuolisation, chromatin condensation, and changes in nucleus and cell shape. Our study showed harmful effects of odorous compounds in chick tissues.

The use of Yucca schidigera and microbial preparation for poultry manure deodorization and hygienization.

The aim of this study was to determine the effectiveness of microbial preparation and Yucca schidigera in the removal of odorous volatile compounds from poultry manure as well as to evaluate antimicrobial properties of these amendments. It was demonstrated that the combined treatment of poultry manure (PM) with the microbial preparation and Y. schidigera extract can reduce the concentration of odorants by 58%-73%, depending on the tested compound. When Y. schidigera extract and the microbial preparation were applied at a time interval of 48 h, the deodorization efficiency was improved by 6-24%. Furthermore, Y. schidigera extract has antimicrobial properties, which affect poultry manure hygienization. It was found that when the microbial preparation was enriched with Lactobacillus plantarum, it became insensitive to the antimicrobial properties of Y. schidigera.

Impact of a microbial-mineral biopreparation on microbial community and deodorization of manures.

The aim of this study was to determine the number of bacteria in poultry, cattle and swine manure in order to perform hygienization and deodorization using a microbial-mineral biopreparation. The highest number of bacteria was recorded in laying hens manure (5.1×10(10) cfu/g). It was noted that bacteria: coliforms, E. coli, Clostridium, Enterococcus number was reduced (1-2 log) after the biopreparation application. The investigated odorous compound concentrations were reduced with 34-78% efficiency, depending on the type of manure and odorant. All odorous compounds were efficiently reduced only in the case of laying hen manure.

Co-digestion of sewage sludge and dewatered residues from enzymatic hydrolysis of sugar beet pulp.

UNLABELLED: Sugar beet pulp residues (SBPR) from hydrolysis and dewatering of beet pulp were co-digested with municipal sewage sludge (MSS). The highest biogas yields of nearly 512 dm(3)/kg VSfed (volatile solids fed) were achieved for SBPR, treated both as the monosubstrate and as a mixture with MSS (1 : 1 by weight). Simultaneously, the highest methane production of 348 dm(3) CH4/kg VSfed was determined when the sewage sludge was co-digested with 35% SBPR. The analysis of digestate showed that neither ammonia nor volatile fatty acids destabilized the biogas production. IMPLICATIONS: Processing of sugar beet pulp into bioethanol via enzymatic hydrolysis and microbial fermentation has become increasingly attractive. However, in this process, only the liquid fraction derived from hydrolysis is subjected into alcoholic fermentation, whereas the remaining solid fraction needs to be utilized. This study demonstrated that sugar beet pulp residues after bioethanol production can successfully be co-digested with sewage sludge to increase biogas productivity of anaerobic digesters located at wastewater treatment plants.

Co-digestion of pig slaughterhouse waste with sewage sludge.

Slaughterhouse wastes (SHW) are potentially very attractive substrates for biogas production. However, mono-digestion of these wastes creates great technological problems associated with the inhibitory effects of ammonia and fatty acids on methanogens as well as with the foaming in the digesters. In the following study, the co-digestion of slaughterhouse wastes with sewage sludge (SS) was undertaken. Batch and semi-continuous experiments were performed at 35°C with municipal sewage sludge and pig SHW composed of meat tissue, intestines, bristles and post-flotation sludge. In batch assays, meat tissue and intestinal wastes gave the highest methane productions of 976 and 826 dm(3)/kg VS, respectively, whereas the methane yield from the sludge was only 370 dm(3)/kg VS. The co-digestion of sewage sludge with 50% SHW (weight basis) provided the methane yield exceeding 600 dm(3)/kg VS, which was more than twice as high as the methane production from sewage sludge alone. However, when the loading rate exceeded 4 kg VS/m(3) d, a slight inhibition of methanogenesis was observed, without affecting the digester stability. The experiments showed that the co-digestion of sewage sludge with large amount of slaughterhouse wastes is feasible, and the enhanced methane production does not affect the digester stability.