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.

Mild alkaline conditions affect digester performance and community dynamics during long-term exposure.

This paper examines the adaptive responses of microbial communities to gradual shifts in pH toward the mild alkaline range in anaerobic digestion (AD) systems. The results indicate that a pH of 8.0 serves as a critical upper limit for stable AD operation, beyond which microbial efficiency declines, underscoring the importance of microbial resilience against elevated pH stress. Specifically, hydrolysis genera, e.g. Eubacterium and Anaerobacterium, and syntrophic bacteria were crucial for reactor stability. Fibrobacter had also been shown to play a key role in the accumulation of propionate, thus leading to its dominance in the volatile fatty acid profile throughout the experimental phases. Overall, this investigation revealed the potential adaptability of microbial communities in AD systems to mild alkaline pH shifts, emphasizing the hydrolysis bacteria and syntrophic bacteria as key factors for maintaining metabolic function in elevated pH conditions.

A redox-based strategy to enhance propionic and butyric acid production during anaerobic fermentation.

This study investigated the selective production of volatile fatty acids (VFAs) during anaerobic mixed-culture fermentation. The experiment used chicken manure (CM) as a potential substrate to produce high added-value propionic acid and butyric acid under an alkaline environment. The conversion of CM into selective VFAs depends highly on operational conditions such as pH and redox balance. Therefore, the current experiment is designed to employ amino acid addition and develop a redox balance control method to control the final VFA profile. This study showed that 0.2-5.0 % valine and threonine addition successfully enhanced propionic acid and butyric acid production during alkaline fermentation and hence decreased the proportion of acetic acid from 83 % to approximately 47 %. The oxidation-reduction potential (ORP) and redox cofactor ratio (NADH/NAD+) were measured to support the selective VFA production mechanism. The results obtained in this study bring extra value to the valorization of CM within the circular economy concept for selective value-added VFA production.

Novel approach to ammonia recovery from anaerobic digestion via side-stream stripping at multiple pH levels.

Ammonia recovery from anaerobic digesters via side-stream stripping is a technique to recover nitrogen from manure wastes. This study demonstrated a novel approach to determining ammonia recovery to maintain total ammonia concentrations in the digester in the range of 1.7-2.1 gN/L. Increasing the pH during stripping from 8, 8.5 to 9.5 did not affect the stability of the digester. Methane yields of 60-80 mL/(gVS.d) and volatile fatty acid concentrations of 0-500 mg/L were reported throughout its operation. The low solubilisation increase upon recirculation of the digestate explained the lack of change in methane yields due to side-stream stripping. Increasing the pH during stripping also did not affect the digester's operating pH, which was attributed to the neutralising effect of biogas as stripping gas. Therefore, total ammonia concentrations in the digester can be controlled by determining the extent of ammonia recovery, and the pH during stripping can be increased without compromising the digester's stability.

Behavior of trace elements and micronutrients in manure digestate during ozone treatment.

Digestate treatment techniques have recently been proposed as a strategy to increase the ultimate biogas yield from dairy manure and to improve the digestate quality as an organic fertilizer. These studies however rarely take the trace elements (TE) and nutrient partitioning into account. This study focusses on ozone treatment (5-40 g O3 kg-1 Total Solids (TS)) as a digestate treatment technique to control the concentration of TE and nutrients in the liquid phase of the digestate. Controlling the TE and nutrient concentrations in the liquid and solid digestate can improve the agronomic value of dairy manure digestate. The ozone concentration of the gas stream entering reactor was 48.53 g O3/Nm³ or 3.4% w/w O3 in O2-gas. The experiments were repeated using pure oxygen gas to investigate its influence. The results from ozonation and oxygenation of the dairy manure digestates revealed that O3 treatment up to 40 g O3 kg-1 TS did not have a more pronounced effect on the biochemical parameters compared to supplementation of pure O2. Ozonation of the digestate and the supernatant showed that the TE concentration in the liquid phase followed a parabolic profile. The observed initial increase in this parabolic profile was explained by the release of TE from the organic matter to the supernatant causing an increase in TE concentration, followed by a decrease due to precipitation of TE as hydroxides and sulfides, due to the increasing pH and sulphur concentrations.

Effect of digestate disintegration on anaerobic digestion of organic waste.

Recently, digestate disintegration gained interest as an alternative strategy to feedstock pretreatment for anaerobic digestion. This study evaluated the effect of three different digestate disintegration methods (hydrogen peroxidation, ozone treatment and ultrasound) on manure digestate, potato waste digestate and mixed organic waste digestate. Lab-scale anaerobic digestion experiments were carried out by adding disintegrated digestate to the related substrate and inoculum with simulated recycle ratios of 0.2 and 0.5. Ultrasound disintegration of potato waste digestate yields 22.5% increase in biogas production. An increase in biogas production was linked to the treated digestate amount and the treatment dosage. First order model was used to investigate the effect of digestate disintegration on the first order reaction rate constant (k). The decrease in k and increase in biogas production were linearly correlated. This correlation was explained by the increased bioavailability of the organic matter and possible negative effects of digestate disintegration on the microorganisms.

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 acid.

Biodegradation of tetracycline under various conditions and effects on microbial community.

Five hundred tons of antibiotics are consumed yearly in the world. In this study, the biodegradation characteristics of tetracycline (TET) under nitrate-reducing, sulfate-reducing, and methanogenic conditions were determined by batch tests. Also, effects of TET on mixed microbial cultures were revealed by microbiological analysis. In this scope, gas generation and composition, dissolved organic carbon, and electron acceptor concentrations were monitored during 120 days. Additionally, changes on quantities of specific microbial groups were determined by Q-PCR. TET showed non-biodegradable behavior under nitrate- and sulfate-reducing conditions, whereas slightly biodegradable behavior under methanogenic conditions approximately 46 % degradation. The effects of TET on the abundance of mixed culture varied according to taxonomic units. Sulfate-reducing bacteria were inhibited by TET, while archaeal, bacterial, and methanogenic populations were not affected significantly.