Biology subjects

De Vrieze, J.

Publications and source records attributed to De Vrieze, J..

11 recordsLinked to original sources

bioRxiv2026

Cultivation-dependent effects of quorum sensing signals on a lactic acid and chain-elongating bacterium

Microbial chain elongation enables the conversion of organic waste into higher-value products and is therefore a promising process for circular biomanufacturing. However, the microbial interactions governing chain elongation communities remain poorly understood. While quorum sensing has been extensively studied in the context of pathogens and model organisms, research on the perception of quorum-sensing molecules by non-model organisms and their effects within microbial consortia has remained limited. Here, Lactiplantibacillus plantarum and Megasphaera elsdenii were selected as representatives of two key functional guilds in chain elongation communities, namely lactic acid bacteria and chain-elongating bacteria. The effects of different exogenous quorum sensing molecules were evaluated in pure cultures and co-cultures using microtiter plates and serum bottles. Both organisms exhibited distinct molecule-dependent responses for both growth and biofilm formation. Moreover, the response of M. elsdenii was highly dependent on the supplied substrate. Despite changes in growth and/or biofilm formation, product yield and product spectra remained largely unaffected. Importantly, responses observed in pure cultures did not predict co-culture behavior, and no clear response to the tested molecules was detected in the co-culture grown in serum bottles. These findings demonstrate that responses to quorum sensing molecules are strongly dependent on the signal, substrate, microbial context, and cultivation conditions. These results highlight the limited predictive power of pure-culture assays for microbial communication in interacting communities and emphasize the importance of studying signal perception under process-relevant cultivation conditions.

microbiology↗

bioRxiv2025

BIO-SPEC: an open-source bench-top parallel bioreactor system.

The BIO-SPEC is an open-source, cost-effective, and modular bench-top bioreactor system designed for batch, sequencing batch, and chemostat cultivation. Featuring thermoelectric condensers to eliminate the need for a chiller, it ensures stable long-term operation. Controlled by a Raspberry Pi, the BIO-SPEC offers flexibility in headplate design, gas supply, and feeding strategies, making it a versatile alternative to high-cost commercial systems. This paper details the design, construction, and validation of the BIO-SPEC system, demonstrating its potential to advance microbiology and bioprocessing research through accessible and reliable hardware at a fraction of the cost.

microbiology↗

bioRxiv2025

Anaerobic co-digestion of sewage sludge and food waste: staging and carriers enhance system performance and process stability

Sewage sludge generated during biological wastewater treatment requires adequate treatment. Anaerobic digestion of sewage sludge is preferred, yet, it is often characterized by low biogas production rates. Co-digestion of sewage sludge with food waste could improve the hydrolysis rate and nutrients balance, resulting in higher biogas production rates. In this study, single- and two-stage anaerobic reactors were operated, with and without carriers in semi-continuous mode for 205 days for assessing the expected improved biogas production rates of sewage sludge and food waste co-digestion. Mono-digestion of sewage sludge resulted in average biogas production rates of 230{+/-}31 and 538{+/-}40 mLbiogas L-1 d-1 for single- and two-stage systems with carriers, respectively. Co-digestion of sewage sludge with food waste resulted in an improved biogas production rates in both systems. Average biogas production rates of 868{+/-}114 and 1507{+/-}174 mLbiogas L-1 d-1 were measured in single- and two-stage systems, respectively, with carriers during co-digestion of food waste (30%) and sewage sludge (70%). In all cases, the addition of carriers improved biogas production, and reduced volatile fatty acid accumulation. In conclusion, two-stage co-digestion of sewage sludge and food waste with carriers achieved the highest biogas production and long-term stability.

bioengineering↗

bioRxiv2024

Molybdate application in the early stages of shrimp growth suppresses sulphide formation in a shrimp pond bottom model

Oxygen depletion and sulphide formation, resulting from the accumulation of organic waste, are common challenges in shrimp ponds that could result in complete harvest failure. The stage at which these circumstances occur during the shrimp growth period remains elusive, yet, knowledge of the timing of oxygen depletion and sulphide formation is essential to enable remediating actions. Here, we used an experimental shrimp pond model at different stages in the shrimp growth period to determine when oxygen depletion and sulphide production occur. Microscale depth measurements of oxygen and H2S were determined using microelectrodes to visualize their profiles at different depths of the water-sediment interface and the sediment. We evaluated the potential of different molybdate concentrations at different stages to determine the optimal conditions to suppress H2S formation. Oxygen depletion and sulphide production took place in the middle of the shrimp growth cycle in the simulated model of waste accumulation. The addition of molybdate was only effective in the early stages of the onset of oxygen depletion and H2S formation, and residual molybdate was required to ensure a continuous suppression of sulphide production. However, oxygen depletion could not be prevented and reintroduction of oxygen did not occur when molybdate was added. In conclusion, molybdate appeared to be an effective strategy to suppress H2S formation at the onset of its production in a shrimp pond bottom model.

microbiology↗

bioRxiv2024

Ectoine production through a marine methanotroph-microalgae culture allows complete biogas valorization

Methanotrophs have recently emerged as a promising platform for producing bio-based chemicals, like ectoine, from biogas, offering an economical alternative to glucose. However, most studies have focused solely on CH4 consumption, often overlooking the CO2, which is both produced by methanotrophs and present in biogas, despite its potential as a carbon source for microorganisms, such as microalgae. In this study, marine methanotrophic-microalgal cultures were enriched from environmental samples collected at the North Sea coast to explore ectoine production from both CH4 and CO2 in biogas. The sediment-derived culture exhibited the highest CH4 removal efficiency and CO2 uptake, and was selected for further experiments. The culture was primarily composed of Methylobacter marinus, Methylophaga marina, and the microalga Picochlorum oklahomensis. Gas consumption, growth, and ectoine production were evaluated under varying salinity levels and osmotic stress. The NaCl concentrations above 6% negatively impacted CH4 oxidation and inhibited ectoine synthesis, while osmotic shocks enhanced ectoine accumulation, with a maximum ectoine content of 51.3 mgectoine gVSS-1 at 4.5% NaCl. This study is the first to report ectoine production from methanotroph-microalgal cultures, showing its potential for biogas valorization into high-value bio-based chemicals, like ectoine, marking a significant step toward sustainable biogas utilization. HighlightsO_LIMethanotroph-microalgae cultures can valorize CH4 and CO2 from biogas into ectoine C_LIO_LINaCl concentrations above 6% reduced CH4 uptake and inhibited ectoine synthesis C_LIO_LIOsmotic shock at 4.5% NaCl enhanced ectoine accumulation C_LIO_LIFirst report of ectoine production using methanotroph-microalgal cultures C_LI

bioengineering↗

bioRxiv2024

Establishing a co-culture aggregate of N-cycle bacteria to elucidate flocculation in biological wastewater treatment

Biological flocculation is a complex phenomenon that is often treated as a black box. As a result, flocculation problems are usually remediated without knowledge of the exact causes. We show that it is feasible to exploit a model (N-cycle) consortium with reduced complexity to fundamentally study bioflocculation. Strong nitrifier microcolonies were formed during oxic/anoxic cycles in sequencing batch reactors, using alginate entrapment as a cell retention system. After release of these aggregates into suspension, macroclusters with flocs of the denitrifier were observed. These results suggest that a living model of a full-scale activated sludge floc can be built through the use of this bottom-up approach. By eliminating shifts in the microbial community, the applied experimental conditions have a more direct effect on the observations.

microbiology↗

bioRxiv2023

Long-term sulphide mitigation through molybdate at shrimp pond bottoms

Shrimp are commonly cultured in earthen aquaculture ponds where organic-rich uneaten feed and faeces accumulate on and in the sediment to form anaerobic zones. Since the pond water is rich in sulphate, these anaerobic conditions eventually lead to the production of sulphide. Sulphides are toxic and even lethal to the shrimp that live on the pond sediment, but physicochemical and microbial reactions that occur during the accumulation of organic waste and the subsequent formation of sulphide in shrimp pond sediments remain unclear. Molybdate treatment is a promising strategy to inhibit sulphate reduction, thus, preventing sulphide accumulation. We used an experimental shrimp pond model to simulate the organic waste accumulation and sulphide formation during the final 61 days of a full shrimp growth cycle. Sodium molybdate (5 and 25 mg/L Na2MoO4.2H2O) was applied as a preventive strategy to control sulphide production before oxygen depletion. Molybdate addition partially mitigated H2S production in the sediment, and delayed its transfer to the bulk liquid by pushing the higher sulphide concentration zone towards deeper sediment layers. Molybdate treatment at 25 mg/L significantly impacted the overall microbial community composition and treated samples (5 and 25 mg/L molybdate) had about 50% higher relative abundance of sulphate reducing bacteria than the control (no molybdate) treatment. In conclusion, molybdate has the potential to work as mitigation strategy against sulphide accumulation in the sediment during shrimp growth by directly steering the microbial community in a shrimp pond system.

bioengineering↗

bioRxiv2021

Triangulation of microbial fingerprinting in anaerobic digestion reveals consistent fingerprinting profiles

The anaerobic digestion microbiome has been puzzling us since the dawn of molecular methods for mixed microbial community analysis. Monitoring of the anaerobic digestion microbiome can either take place via a non-targeted holistic evaluation of the microbial community through fingerprinting or by targeted monitoring of selected taxa. Here, we compared four different microbial community fingerprinting methods, i.e., amplicon sequencing, metaproteomics, metabolomics and cytomics, in their ability to characterise the full-scale anaerobic digestion microbiome. Cytometric fingerprinting through cytomics reflects a, for anaerobic digestion, novel, single cell-based approach of direct microbial community fingerprinting by flow cytometry. Three different digester types, i.e., sludge digesters, digesters treating agro-industrial waste and dry anaerobic digesters, each reflected different operational parameters. The -diversity analysis yielded inconsistent results, especially for richness, across the different methods. In contrast, {beta}-diversity analysis resulted in comparable profiles, even when translated into phyla or functions, with clear separation of the three digester types. In-depth analysis of each methods features i.e., operational taxonomic units, metaproteins, metabolites, and cytometric traits, yielded certain similar features, yet, also some clear differences between the different methods, which was related to the complexity of the anaerobic digestion process. In conclusion, cytometric fingerprinting through flow cytometry is a reliable, fast method for holistic monitoring of the anaerobic digestion microbiome, and the complementary identification of key features through other methods could give rise to a direct interpretation of anaerobic digestion process performance.

bioengineering↗

bioRxiv2020

Sterilizing Immunity against SARS-CoV-2 Infection in Mice by a Single-Shot and Modified Imidazoquinoline TLR7/8 Agonist-Adjuvanted Recombinant Spike Protein Vaccine

The search for vaccines that protect from severe morbidity and mortality as a result of infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes coronavirus disease 2019 (COVID-19) is a race against the clock and the virus. Several vaccine candidates are currently being tested in the clinic. Inactivated virus and recombinant protein vaccines can be safe options but may require adjuvants to induce robust immune responses efficiently. In this work we describe the use of a novel amphiphilic imidazoquinoline (IMDQ-PEG-CHOL) TLR7/8 adjuvant, consisting of an imidazoquinoline conjugated to the chain end of a cholesterol-poly(ethylene glycol) macromolecular amphiphile). This amphiphile is water soluble and exhibits massive translocation to lymph nodes upon local administration, likely through binding to albumin. IMDQ-PEG-CHOL is used to induce a protective immune response against SARS-CoV-2 after single vaccination with trimeric recombinant SARS-CoV-2 spike protein in the BALB/c mouse model. Inclusion of amphiphilic IMDQ-PEG-CHOL in the SARS-CoV-2 spike vaccine formulation resulted in enhanced immune cell recruitment and activation in the draining lymph node. IMDQ-PEG-CHOL has a better safety profile compared to native soluble IMDQ as the former induces a more localized immune response upon local injection, preventing systemic inflammation. Moreover, IMDQ-PEG-CHOL adjuvanted vaccine induced enhanced ELISA and in vitro microneutralization titers, and a more balanced IgG2a/IgG1 response. To correlate vaccine responses with control of virus replication in vivo, vaccinated mice were challenged with SARS-CoV-2 virus after being sensitized by intranasal adenovirus-mediated expression of the human angiotensin converting enzyme 2 (ACE2) gene. Animals vaccinated with trimeric recombinant spike protein vaccine without adjuvant had lung virus titers comparable to non-vaccinated control mice, whereas animals vaccinated with IMDQ-PEG-CHOL-adjuvanted vaccine controlled viral replication and infectious viruses could not be recovered from their lungs at day 4 post infection. In order to test whether IMDQ-PEG-CHOL could also be used to adjuvant vaccines currently licensed for use in humans, proof of concept was also provided by using the same IMDQ-PEG-CHOL to adjuvant human quadrivalent inactivated influenza virus split vaccine, which resulted in enhanced hemagglutination inhibition titers and a more balanced IgG2a/IgG1 antibody response. Enhanced influenza vaccine responses correlated with better virus control when mice were given a lethal influenza virus challenge. Our results underscore the potential use of IMDQ-PEG-CHOL as an adjuvant to achieve protection after single immunization with recombinant protein and inactivated vaccines against respiratory viruses, such as SARS-CoV-2 and influenza viruses.

microbiology↗

bioRxiv2019

The feedstock microbiome selectively steers process stability during the anaerobic digestion of waste activated sludge

Strategies to enhance process performance of anaerobic digestion remain of key importance to achieve further spreading of this technology for integrated resource recovery from organic waste streams. Continuous inoculation of the microbial community in the digester via the feedstock could be such a cost-effective strategy. Here, anaerobic digestion of fresh waste activated sludge (WAS) was compared with sterilized WAS in response to two common process disturbances, i.e., organic overloading and increasing levels of salts, to determine the importance of feedstock inoculation. A pulse in the organic loading rate severely impacted process performance of the digesters fed sterile WAS, with a 92 {+/-} 45 % decrease in methane production, compared to a 42 {+/-} 31 % increase in the digesters fed fresh WAS, relative to methane production before the pulse. Increasing salt pulses did not show a clear difference in process performance between the digesters fed fresh and sterile WAS, and process recovery was obtained even at the highest salt pulse of 25 g Na+ L-1. Feedstock sterilisation strongly impacted the microbial community in the digesters. In conclusion, feedstock inoculation can be considered a cheap, yet, disturbance-specific strategy to enhance process stability in full-scale anaerobic digestion processes.

bioengineering↗

bioRxiv2019

Exploiting the unwanted: sulphate reduction enables phosphate recovery from energy-rich sludge during anaerobic digestion.

Anaerobic digestion is shifting from a single-purpose technology for renewable energy recovery from organic waste streams to a process for integrated resource recovery. The valorisation of high-rate energy- and phosphorus-rich sludge creates the opportunity for their combined recovery. This phosphate is present in a precipitated form in the sludge, and its release into the liquid phase is an important issue before recovery can be achieved. The objective of this research was to exploit the \"unwanted\" sulphate reduction process for the release of phosphate into the liquid phase during anaerobic digestion, thus, making it available for recovery. Two different treatments were considered, i.e., a control digester and a digester to which sulphate was added, each operated in triplicate for a period of 119 days. The control digester showed stable methane production at 628 {+/-} 103 mL CH4 L-1 d-1, with a feedstock COD (chemical oxygen demand) conversion efficiency of 89.5 {+/-} 14.6 %. In contrast, the digester with sulphate addition showed a 29.9 {+/-} 15.3 % decrease in methane production, reaching an \"inhibited steady state\", but phosphate release into the liquid phase increased with a factor 4.5, compared to the control digester. This inhibited steady state coincided with a clear shift from a Methanosaetaceae to a Methanosarcinaceae dominated methanogenic community. Overall, the sulphate reduction process allows phosphate release during the anaerobic digestion process, yet, at the cost of a reduced methane production rate.

bioengineering↗