Methanogenic capacity and robustness of hydrogenotrophic cultures based on closed nutrient recycling via microbial catabolism: Impact of temperature and microbial attachment.

A biological methanation system based on nutrient recycling via mixed culture microbial catabolism was investigated at mesophilic (37 °C) and thermophilic (55 °C) temperatures. At mesophilic temperatures, the formation of biofilms on two different types of material was assessed. Results showed that with intense mixing the biofilm reactors presented methanogenic capacities (per working volume) 50% higher than the ones operated with suspended cultures. Gas feeding rates of 200 L/L/d were achieved at a H2/CO2 to CH4 conversion efficiency of above 90% by linking two reactors in series. Furthermore the robustness of the cultures was assessed under a series of inhibitory conditions that simulated possible process interferences at full scale operation. Full recovery after separate intense oxygenation and long starvation periods was observed within 2-5 days.

Enhancement of microbial density and methane production in advanced anaerobic digestion of secondary sewage sludge by continuous removal of ammonia.

Ammonia inhibition mitigation in anaerobic digestion of high solids content of thermally hydrolysed secondary sewage sludge by the NH4+ affinitive clinoptilolite and a strong acid type ion-exchange resin S957 was investigated. Continuous NH4+-N removal was achieved through ion-exchanging at both temperatures with average removals of 50 and 70% for the clinoptilolite and resin dosed reactors, respectively. Approximate 0.2-0.5unit of pH reduction was also observed in the dosed reactors. The synergy of NH4+-N removal and pH reduction exponentially decreased free NH3 concentration, from 600 to 90mg/L at 43°C, which mitigated ammonia inhibition and improved methane yields by approximately 54%. Microbial community profiling suggested that facilitated by ammonia removal, the improvement in methane production was mainly achieved through the doubling in bacterial density and a 6-fold increase in population of the Methanosarcinaceae family, which in turn improved the degradation of residual volatile fatty acids, proteins and carbohydrates.

Third generation poly(hydroxyacid) composite scaffolds for tissue engineering.

Bone tissue engineering based on scaffolds is quite a complex process as a whole gamut of criteria needs to be satisfied to promote cellular attachment, proliferation and differentiation: biocompatibility, right surface properties, adequate mechanical performance, controlled bioresorbability, osteoconductivity, angiogenic cues, and vascularization. Third generation scaffolds are more of composite types to maximize biological-mechanical-chemical properties. In the present review, our focus is on the performance of micro-organism-derived polyhydroxyalkanoates (PHAs)-polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-valerate (PHBV)-composite scaffolds with ceramics and natural polymers for tissue engineering applications with emphasis on bone tissue. We particularly emphasize on how material properties of the composites affect scaffold performance. PHA-based composites have demonstrated their biocompatibility with a range of tissues and their capacity to induce osteogenesis due to their piezoelectric properties. Electrospun PHB/PHBV fiber mesh in combination with human adipose tissue-derived stem cells (hASCs) were shown to improve vascularization in engineered bone tissues. For nerve and skin tissue engineering applications, natural polymers such as collagen and chitosan remain the gold standard but there is scope for development of scaffolds combining PHAs with other natural polymers which can address some of the limitations such as brittleness, lack of bioactivity and slow degradation rate presented by the latter. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1667-1684, 2017.

Closed nutrient recycling via microbial catabolism in an eco-engineered self regenerating mixed anaerobic microbiome for hydrogenotrophic methanogenesis.

A novel eco-engineered mixed anaerobic culture was successfully demonstrated for the first time to be capable of continuous regeneration in nutrient limiting conditions. Microbial catabolism has been found to support a closed system of nutrients able to enrich a culture of lithotrophic methanogens and provide microbial cell recycling. After enrichment, the hydrogenotrophic species was the dominating methanogens while a bacterial substratum was responsible for the redistribution of nutrients. q-PCR results indicated that 7% of the total population was responsible for the direct conversion of the gases. The efficiency of H2/CO2 conversion to CH4 reached 100% at a gassing rate of above 60v/v/d. The pH of the culture media was effectively sustained at optimal levels (pH 7-8) through a buffering system created by the dissolved CO2. The novel approach can reduce the process nutrient/metal requirement and enhance the environmental and financial performance of hydrogenotrophic methanogenesis for renewable energy storage.

Volatile fatty acids platform from thermally hydrolysed secondary sewage sludge enhanced through recovered micronutrients from digested sludge.

The extracellular polymeric substances and microbial cytoplasmic contents seem to hold inorganic ions and organic products, such as proteins and carbohydrates that are of critical importance for the metabolism of hydrolytic and acidogenic anaerobic microorganisms. The addition of soluble microbially recovered nutrients from thermally treated digestate sludge, for the fermentation of thermally hydrolysed waste activated sludge, resulted in higher volatile fatty acids yields (VFAs). The yield of VFAs obtained from the recovered microbial nutrients was 27% higher than the no micronutrients control, and comparable to the yield obtained using a micronutrients commercial recipe. In addition, the use of a low pH resulting from a high sucrose dose to select spore forming acidogenic bacteria was effective for VFA production, and yielded 20% higher VFAs than without the pH shock and this associated with the addition of recovered microbial nutrients would overcome the need to thermally pre-treat the inoculum.

The use of NaCl addition for the improvement of polyhydroxyalkanoate production by Cupriavidus necator.

External stress factors in the form of ionic species or temperature increases have been shown to produce a stress response leading to enhanced PHA production. The effect of five different NaCl concentrations, namely 3.5, 6.5, 9, 12 and 15 g/l NaCl on PHA productivity using Cupriavidus necator has been investigated alongside a control (no added NaCl). A dielectric spectroscopy probe was used to measure PHA accumulation online in conjunction with the chemical offline analysis of PHA. The highest PHA production was obtained with the addition of 9 g/l NaCl, which yielded 30% higher PHA than the control. Increasing the addition of NaCl to 15 g/l was found to inhibit the production of PHA. NaCl addition can therefore be used as a simple, low cost, sustainable, non toxic and non reactive external stress strategy for increasing PHA productivity.

Increasing polyhydroxyalkanoate (PHA) yields from Cupriavidus necator by using filtered digestate liquors.

The production of polyhydroxyalkanoates (PHAs) using digestate liquor as culture media is a novel application to extend the existing uses of digestates. In this study, two micro-filtered digestates (0.22 µm) were evaluated as a source of complex culture media for the production of PHA by Cupriavidus necator as compared to a conventional media. Culture media using a mixture of micro-filtered liquors from food waste and from wheat feed digesters showed a maximum PHA accumulation of 12.29 g/l PHA, with 90% cell dry weight and a yield of 0.48 g PHA/g VFA consumed, the highest reported to date for C. necator studies. From the analysis of the starting and residual media, it was concluded that ammonia, potassium, magnesium, sulfate and phosphate provided in the digestate liquors were vital for the initial growth of C. necator whereas copper, iron and nickel may have played a significant role in PHA accumulation.

Integration of NIRS and PCA techniques for the process monitoring of a sewage sludge anaerobic digester.

This study investigates the use of Hotelling's T(2) control charts as the basis of a process monitor for sewage sludge anaerobic digestion. Fourier transform near infrared spectroscopy was used to produce partial least squares regression models of volatile fatty acids, bicarbonate alkalinity and volatile solids. These were utilised in a series of principle component analysis models along with spectral data from digestate and feedstock samples to produce a pseudo steady state model, which was then used with an independent test set to evaluate the system. The system was able to identify disturbances to the digester due to a temporary alteration of the type of feedstock to the digester and separately, halving of the hydraulic retention time of the digester. It could also provide advance warning of disturbances to the digester. This technique could be used to improve the performance of sewage sludge anaerobic digesters by enabling optimisation of the process.

Addressing the challenge of optimum polyhydroxyalkanoate harvesting: monitoring real time process kinetics and biopolymer accumulation using dielectric spectroscopy.

In this study, dielectric spectroscopy was utilised to evaluate and define the optimum harvesting time for polyhydroxyalkanoates (PHA) production. It is essential to harvest PHA at the optimum time during fermentation for maximum yield, otherwise cells start degrading. Two carbon sources (acetic and butyric acids) were used in laboratory based experiments and a number of samples were measured ex situ for PHA production. The real-time measured capacitance in addition of identifying the cells growth phase, it correlated very well with ex situ measured PHA produced within the cells. The probe has proven to be a useful tool to assess process kinetics, to monitor real-time cell growth, PHA produced and defining the optimum harvesting time.