Biology subjects

Thi, S. S.

Publications and source records attributed to Thi, S. S..

5 recordsLinked to original sources

bioRxiv2023

Microbial community-based bacterial protein from soybean-processing wastewater as a value-added alternative fish feed ingredient for Asian sea bass (Lates calcarifer)

As the global demand for food increases, aquaculture plays a key role as the fastest growing animal protein sector. However, existing aquafeeds contain protein ingredients that are not sustainable under current production systems. We evaluated the use of microbial community-based single cell protein (SCP), produced from soybean processing wastewater, as a partial fishmeal protein substitute in juvenile Asian seabass (Lates calcarifer). A 24-day feeding trial was conducted with a control fishmeal diet and a 50% fishmeal replacement with microbial community-based SCP as an experimental group, in triplicate tanks containing 20 fish each. Both diets met the protein, essential amino acids (except for lysine), and fat requirements for juvenile Asian sea bass. The microbial composition of the SCP was dominated by the genera Acidipropionibacterium and Propioniciclava, which have potential as probiotics and producers of valuable metabolites. The growth performance in terms of percent weight gain, feed conversion ratio (FCR), specific growth rate (SGR), and survival were not significantly different between groups after 24 days. The experimental group had less variability in terms of weight gain and FCR than the control group. Overall, microbial community-based protein produced from soybean processing wastewater has potential as a value-added feed ingredient for sustainable aquaculture feeds.

microbiology↗

bioRxiv2022

Microbial community-based production of single cell protein from soybean-processing wastewater of variable chemical composition

The use of food-processing wastewaters to produce microbial biomass-derived single cell protein (SCP) is a sustainable way to meet the global food demand. Yet, despite the potential benefits of lower costs and greater resource recovery compared to pure cultures, bioconversion processes relying on microbial community-based approaches to SCP production have received scarce attention. Here, we evaluated SCP production from soybean-processing wastewaters under controlled reactor conditions using the existent microbial communities in these wastewaters. Six sequencing batch reactors of 4.5-L working volume were operated at 30 {square} for 34 d in cycles consisting of 3-h anaerobic and 9-h aerobic phases. Four reactors received no microbial inoculum and the remaining two were amended with a 1.5 L of mixed culture from a prior microbial community-based SCP production. Microbial characterization was done via 16S rRNA gene metabarcoding. Influent wastewater batches had variable chemical characteristics but a similar microbial composition. Reactors produced more SCP when fed with wastewaters of higher soluble total Kjeldahl nitrogen (sTKN) content and a lower carbon-to-nitrogen ratio (sCOD:sTKN). The biomass protein yield ranged from 0.24 to 3.13 g protein/g sTKN, with a maximum protein content of 50%. An average of 92% of sCOD and 73% of sTN removal was achieved. Distinct microbial communities were enriched in all six bioreactors after 34 d, where the prevailing genera included Azospirillum, Rhodobacter, Lactococcus, Novosphingobium, and Acidipropionibacterium. In contrast, the microbial community of influent wastewaters was dominated by Lactococcus and Weissella. We showed that constituents in soybean wastewater can be converted to SCP through microbial community-based growth processes and demonstrated the effect of variable influent wastewater composition on SCP production.

microbiology↗

bioRxiv2021

Global warming readiness: Feasibility of enhanced biological phosphorus removal from wastewater at 35oC

Recent research has shown enhanced biological phosphorus removal (EBPR) from municipal wastewater at warmer temperatures around 30{degrees}C to be stable in both laboratory-scale reactors and full-scale treatment plants. In the context of a changing climate, the feasibility of EBPR at even higher temperatures is of interest. We operated two lab-scale EBPR sequencing batch reactors with alternating anaerobic and aerobic phases for over 300 days at 30{degrees}C and 35{degrees}C, respectively, and followed the dynamics of the communities of phosphorus accumulating organisms (PAOs) and competing glycogen accumulating organisms (GAOs) using a combination of 16S rRNA gene metabarcoding, quantitative PCR and fluorescent in-situ hybridization analyses. Stable and nearly complete P removal was achieved at 30{degrees}C; similarly, long term P removal was stable at 35{degrees}C with effluent PO43--P concentrations < 0.5 mg/L on half of all monitored days. Diverse and abundant Ca. Accumulibacter amplicon sequence variants were closely related to those found in temperate environments, suggesting that EBPR at this temperature does not require a highly specialized PAO community. The slow-feeding strategy used effectively limited the carbon uptake rates of GAOs, allowing PAOs to outcompete GAOs at both temperatures. Candidatus Competibacter was the main GAO, along with cluster III Defluviicoccus members. These organisms withstood the slow-feeding regime, suggesting that their bioenergetic characteristics of carbon uptake differ from those of their tetrad-forming relatives. This specific lineage of GAOs warrants further study to establish how complete P removal can be maintained. Comparative cycle studies at two temperatures for each reactor revealed higher activity of Ca. Accumulibacter when the temperature was increased from 30{degrees}C to 35{degrees}C, suggesting that the stress was a result of the higher carbon (and/or P) metabolic rates of PAOs and GAOs, the resultant carbon deficiency, and additional community competition. An increase in the TOC to PO43--P ratio (from 25:1 to 40:1) effectively eased the carbon deficiency and benefited the proliferation of PAOs. In general, the slow-feeding strategy and sufficiently high carbon input benefited a high and stable EBPR at elevated temperature and represent basic conditions for full-scale applications.

microbiology↗

bioRxiv2020

Species level enrichment of AnAOB and associated growth morphology under the effect of key metabolites

Nineteen anaerobic ammonium oxidizing bacteria (AnAOB) species have been identified, yet the environmental factors that select for each species within a specialized ecological niche have not been described. We enriched AnAOB from a single inoculum under standard enrichment conditions (reactor R1) with stepwise increase in nitrite and ammonia concentration, nitric oxide (NO) supplementation (reactor R2), or with complex organic carbon using wastewater collected from mainstream wastewater treatment facility (reactor R3). AnAOB were enriched up to 80%, 90% and 50% relative abundance in R1, R2 and R3 respectively. Candidatus Brocadia caroliniensis predominated in all reactors, but a shift towards Ca. Brocadia sinica was consistently observed with increasing ammonium and nitrite concentrations beyond 270 mg NH4-N L-1 and 340 mg NO2-N L-1, respectively. In the presence of NO, growth of heterotrophs were inhibited, and Ca. Jettenia could coexist with Ca. B. caroliniensis before diminishing when nitrite increased to 160 mg NO2-N L-1. In contrast, supplementation of organic carbon led to the emergence of heterotrophic communities that coevolved with Ca. B. caroliniensis. Ca. B. caroliniensis and Ca. Jettenia preferentially form biofilms on reactor surfaces, whereas Ca. Brocadia sinica forms granules in suspension. Our results thus indicate that multiple AnAOB species co-exist and occupy sub-niches in anaerobic ammonium oxidation reactors, that the dominant population can be reversibly shifted by, for example, changing the nitrogen load (i.e. high nitrite concentration favors Ca. Brocadia caroliniensis), and that speciation has implications for wastewater process design, with the optimum cell immobilization strategy (i.e. carriers vs granules) dependent on which species dominates. IMPORTANCEThis study demonstrates how to reversibly and predictably shift dominant anammox population using operating parameters (e.g. high nitrite concentration favours Ca. Brocadia sinica), and that species selection has implications for wastewater process design, illustrated here in terms of dependence of optimum cell immobilization strategy (i.e. carriers vs granules) on which species dominates. The research informs the characterization of AnAOBs at species level as well process design and control strategies targeting Anammox species population dynamics in full scale waste water treatment systems.

ecology↗

bioRxiv2019

Isolation of a putative S-layer protein from anammox biofilm extracellular matrix using ionic liquid extraction

Anaerobic ammonium oxidation (anammox) performing bacteria self-assemble into compact biofilms by expressing extracellular polymeric substances (EPS). Anammox EPS are poorly characterized, largely due to their low solubility in typical aqueous solvents. Pronase digestion achieved 19.5 {+/-} 0.9 and 41.4 {+/-} 1.4% (w/w) more solubilization of Candidatus Brocadia sinica-enriched anammox granules than DNase and amylase respectively. Nuclear magnetic resonance profiling of the granules confirmed that proteins were dominant. We applied ionic liquid (IL) 1-ethyl-3-methylimidazolium acetate and N,N- dimethylacetamide (EMIM-Ac/DMAc) mixture to extract the major structural proteins. Further treatment by anion exchange chromatography isolated homologous S/T-rich proteins BROSI_A1236 and UZ01_01563, which were major components of the extracted proteins and sequentially highly similar to putative anammox surface-layer (S-layer) protein KUSTD1514. EMIM-Ac/DMAc extraction enriched for these putative S-layer proteins against all other major proteins, along with six monosaccharides (i.e. arabinose, xylose, rhamnose, fucose, galactose and mannose). The sugars, however, contributed <0.5% (w/w) of total granular biomass, and were likely co-enriched as glycoprotein appendages. This study demonstrates that S-layer proteins are major constituents of anammox biofilms and can be isolated from the matrix using an ionic liquid-based solvent.

bioengineering↗