Mitigation of Humic Acid Inhibition in Anaerobic Digestion of Cellulose by Addition of Various Salts

Humic compounds are inhibitory to the anaerobic hydrolysis of cellulosic biomass. In this study, the impact of salt addition to mitigate the inhibitory effects of humic compounds was investigated. The experiment was conducted using batch tests to monitor the anaerobic hydrolysis of cellulose in the presence of humic acid. Sodium, potassium, calcium, magnesium and iron salts were tested separately for their efficiency to mitigate humic acid inhibition. All experiments were done under mesophilic conditions (30 °C) and at pH 7. Methane production was monitored online, using the Automatic Methane Potential Test System. Methane production, soluble chemical oxygen demand and volatile fatty acid content of the samples were measured to calculate the hydrolysis efficiencies. Addition of magnesium, calcium and iron salts clearly mitigated the inhibitory effects of humic acid and hydrolysis efficiencies reached up to 75%, 65% and 72%, respectively, which were similar to control experiments. Conversely, potassium and sodium salts addition did not mitigate the inhibition and hydrolysis efficiencies were found to be less than 40%. Mitigation of humic acid inhibition via salt addition was also validated by inductively coupled plasma atomic emission spectroscopy analyses, which showed the binding capacity of different cations to humic aci

Methanotrophy under extreme conditions : biochemistry and physiology of Methylacidiphilum fumariolicum SolV

Methane (CH4) is an important fossil fuel for households and industry, but also a trace gas in the atmosphere. Global warming is a worldwide concern and therefore it is important to increase knowledge of sources and sinks for methane. Fundamental knowledge about methanotrophic bacteria is important since they are sinks for methane diffusing from both biogenic and abiogenic sources. Until 2007, all genera of aerobic methanotrophs could be phylogenetically placed into the Alphaproteobacteria and Gammaproteobacteria. Intrigued by the observations that volcanic regions, characterized by a pH down to 1.8 and temperatures of 50-95 °C, may act as sinks for methane, recently (2007-2008) three research groups independently isolated obligate aerobic methanotrophs from these hostile environments in pure culture. Inocula were obtained from the Solfatara at Pozzuoli near Naples (Italy), Hell's Gate, Tikitere (New Zealand) and the Uzon Caldera, Kamchatka, (Russia). Excitingly, based on 16S rRNA gene sequences all three isolates (strains SolV, V4 and Kam1) could be identified as members of the Verrucomicrobia phylum. This is the first time that the widely distributed Verrucomicrobia phylum, from which most members remain uncultivated, is coupled to a geochemical cycle. This thesis focuses on one of these novel methanotrophs, e.g. Methylacidiphilum fumariolicum SolV isolated using mud from the central mud pool (Fangaia) of the Solfatara near Naples. Initial analyses of strain SolV showed major differences with the classical proteobacterial methanotrophs, e.g. extreme acid tolerance, absence of typical membrane structures, distinct enzymes of the methane oxidation and carbon fixation pathways. This was validated by combined approaches like genomics, electron microscopy, mRNA analyses, 13C-labeling and in this way the metabolism of strain SolV was unraveled in some details. This fundamental knowledge is necessary to assess the environmental importance of these microbes as a methane sink in volcanic areas

Metabolic regulation of “ca. Methylacidiphilum fumariolicum” solv cells grown under different nitrogen and oxygen limitations

Contains fulltext : 103525.pdf (preprint version ) (Open Access

Genomic and physiological analysis of carbon storage in the verrucomicrobial methanotroph "Ca. Methylacidiphilum fumariolicum" SolV

Contains fulltext : 126468.pdf (publisher's version ) (Open Access

Unravelling the protein preference of aquatic worms during waste activated sludge degradation

Worm predation (WP) by Tubifex tubifex was investigated using waste activated sludge (WAS) as the substrate. In order to better understand the sludge degradation mechanisms during WP, the activity of five common hydrolytic enzymes was determined and compared among the initial feed activated sludge, endogenous respirated sludge and worm predated sludge. The results showed that the enzymatic activity decreased upon aerobic (worm) treatment of WAS and that this activity was predominantly associated with the removed solids fraction of the sludge. Interestingly, the protease activity showed a smaller decrease in activity when the worms were present. Flow cell cytometry revealed the release of intestinal bacteria from the worms, which are presumed to be largely responsible for the observed protease activity. Additionally, experiments in which T. tubifex were treated with antibiotics showed that the worms are responsible for a maximum of 73% of the observed proteolytic activity. The remaining 27% is attributed to the intestinal bacteria that exhibit a synergistic relationship with T. tubifex towards protein hydrolysis.Sanitary Engineerin

The biodegradability of aquatic worm predated waste activated sludge: A sequential aerobic and anaerobic treatment approach

The objective of this study was to investigate the effect of waste activated sludge (WAS) predation by the aquatic worm Tubifex tubifex (T. tubifex) on the overall biodegradability of WAS. The initial WAS biodegradability potential was determined in 80 days sequential batch-fed anaerobic and aerobic treatment combinations. These treatment combinations were used as a reference for comparison with the effect of 5-day predation and 40-day anaerobic treatment combinations. Predation and the subsequent anaerobic digestion of the predated solids shows superior solids removal and superior overall conversion rates compared to solely conventional anaerobic digestion. Strikingly, the predation and anaerobic treatment combinations reached the same chemical oxygen demand (COD) and volatile solids (VS) reduction as the reference processes, i.e. 58% and 49% for COD and VS, respectively. Our results show that predation and anaerobic treatment combinations increase solids removal rates, but do not alter the overall biodegradability potential of WAS.</p

Elucidating the microbial community associated with the protein preference of sludge-degrading worms

Sludge predation by aquatic worms results in an increased sludge reduction rate, which is mainly due to the specific removal of a protein fraction from the sludge. As microorganisms play an essential role in sludge hydrolysis a better understanding of the microbial community involved in the worm predation process will provide more insight into the relations between the aquatic worms, their associated microbiome and the efficient sludge reduction. In this study, the microbial community associated with predation by the Tubifex tubifex was investigated. The microbial diversity in the samples of the worm faeces (WF), predated activated sludge and protein-rich substrates were compared. The results indicated that predation on sludge resulted in a microbial change from Actinobacteria (44%) in the sludge, to Proteobacteria (64%) and Bacteriodites (36%) in the WF. Interestingly, the faecal microbial community was more related to the community in (predated) protein-rich substrates than to the community in predated or endogenously respirated activated sludge samples. This similar microbial community could be due to microbial utilisation of protein hydrolysis products. Alternatively, conditions in the worm gut could facilitate a protein hydrolysing community which assists in protein hydrolysis. The genera Burkholderiales, Chryseobacterium and Flavobacterium were found to be associated with predation by T. tubifex.Sanitary Engineerin

Physical and biochemical changes in sludge upon Tubifex tubifex predation

Worm predation (WP) on activated sludge leads to increased sludge degradation rates, irrespective of the type of worm used or reactor conditions employed. However, the cause of the increased sludge degradation rates remains unknown. This paper presents a comparative analysis of the physical and biochemical aspects of predated sludge, providing insight into the hydrolytic mechanisms underlying WP. To this end, the sessile worm Tubifex tubifex was used as a model oligochaete and was batch cultivated in an 18-L airlift reactor. Predation on activated sludge showed an average reduction rate of 12 ± 3.8%/d versus 2 ± 1.3%/d for endogenous respirated sludge. Sludge predation resulted in an increased release of inorganic nitrogen, phosphate and soluble chemical oxygen demand (sCOD). The sCOD consisted mainly of polysaccharides; however, fluorescence excitation emission matrix spectroscopy analysis also revealed the presence of Tryptophan-protein-like substances. Results suggest that the released polysaccharides contain a protein-like element. Additionally, soluble iron increased slightly in concentration after WP. The extent of hydrolysis seemed to reach an average plateau of about 40% volatile solids (VS) reduction after 4 days, which is substantially higher than the 29% VS reduction for endogenous decay of activated sludge after 30 days. Furthermore, T. tubifex predominantly consumed the protein fraction of the extracellular polymeric substances. Results suggest that that the worms specifically target a fraction of the sludge that is predominantly biodegradable under aerobic conditions, albeit at significantly higher degradation rates when compared to the endogenous decay of waste activated sludge.Sanitary Engineerin

Draft genome sequence of the volcano-inhabiting thermoacidophilic methanotroph methylacidiphilum fumariolicum strain solv

Contains fulltext : 93817.pdf (author's version ) (Open Access

Effect of humic acid on anaerobic digestion of cellulose and xylan in completely stirred tank reactors: inhibitory effect, mitigation of the inhibition and the dynamics of the microbial communities.

Inhibition effect of humic acid (HA) on anaerobic digestion of cellulose and xylan and the mitigation potential of the inhibition were evaluated in controlled fed batch reactors at 30 °C and a hydraulic retention time (HRT) of 20 days. Reactor performances were evaluated by biogas production and metabolite measurements for 220 days. Microbial population dynamics of the reactors were monitored with next-generation 16S rRNA gene sequencing at nine different sampling times. Our results showed that increasing levels of HA inhibited the hydrolysis efficiency of the digestion by 40% and concomitantly reduced the methane yield. Addition of hydrolytic enzymes helped to reverse the negative effects of HA, whereas calcium addition did not reverse HA inhibition. Microbiological analyses showed that the relative abundance of hydrolytic/fermentative bacterial groups such as Clostridiales, Bacteroidales and Anaerolineales was significantly lowered by the presence of HA. HA also affected the archaeal populations. Mostly hydrogenotrophic methanogens were negatively affected by HA. The relative abundance of Methanobacteriaceae, Methanomicrobiales-WCHA208 and Unassigned Thermoplasmata WCHA1-57 were negatively affected by the presence of HA, whereas Methanosaetacea was not affected.Sanitary Engineerin