Application of Computational Tools to Support Cooperative Learning in Bioreactor Design Course

Conventional method of teaching Bioreactor Design course are mostly conducted in a teacher-centred manner. This method is inefficient solution for education as compared to more active learning styles which is proven to be more effective in ensuring students to fully comprehend a particular subject. The work presents the use of various computational tools to support the implementation of coopera-tive learning (CL) methods in Analysis and Design of Bioreactor course. This subject is offered to 3rd year students of Chemical-Bioprocess Engineering pro-gram in Universiti Teknologi Malaysia. The CL method was implemented to im-prove student cognitive skills attainment in each of the course learning outcomes. Achievements of student cognitive skills were assessed quantitatively where else effectiveness of the CL method applied were evaluated qualitatively. Results showed that the student performance and attainment of their cognitive skills at thinking level of ‘application’ has improved at least by 30-40%. Reflection analy-sis from students indicated that the proposed student-centred teaching method managed to not only increase students understanding on the subject but also nurtured students creativity and enhances their computational skills.

Direct evidence for deterministic assembly of bacterial communities in full-scale municipal wastewater treatment facilities

In this study we investigated whether bacterial community composition in full-scale wastewater treatment bioreactors can be better explained by niche- or neutral- based theory (deterministic or stochastic) and whether bioreactor design (continuous-flow vs. fill-and-draw) affected community assembly. Four wastewater treatment facilities (one with quadruplicated continuous-flow bioreactors, two with one continuous-flow bioreactor each, one with triplicated fill-and-draw bioreactors) were investigated. Bioreactor community composition was characterized by sequencing of PCR-amplified 16S rRNA gene fragments. Replicate bioreactors at the same wastewater treatment facility had largely reproducible (i.e., deterministic) bacterial community composition, although bacterial community composition in continuous-flow bioreactors was significantly more reproducible ( P < 0.001) than in fill-and-draw bioreactors (Bray-Curtis dissimilarity: μ = 0.48 ± 0.06 vs. 0.58 ± 0.08). Next, we compared our results to previously-used indirect methods for distinguishing between deterministic and stochastic community assembly mechanisms. Synchronicity was observed in the bacterial community composition among bioreactors within the same metropolitan region, consistent with deterministic community assembly. Similarly, a null model-based analysis also indicated that all wastewater bioreactor communities were controlled by deterministic factors and that continuous-flow bioreactors were significantly more deterministic ( P < 0.001) than fill-and-draw bioreactors (nearest-taxon index: μ = 3.3 ± 0.6 vs. 2.7 ± 0.8). Our results indicate that bacterial community composition in wastewater treatment bioreactors is better explained by deterministic community assembly theory; simultaneously, our results validate previously-used but indirect methods to quantify whether microbial communities were assembled via deterministic or stochastic mechanisms. IMPORTANCE Understanding the mechanisms of bacterial community assembly is one of the grand challenges of microbial ecology. In environmental systems, this challenge is exacerbated because “replicate” experiments are typically impossible; that is, microbial ecologists cannot fabricate multiple field-scale experiments of identical, natural ecosystems. Our results directly demonstrate that deterministic mechanisms are more prominent than stochastic mechanisms in the assembly of wastewater treatment bioreactor communities. Our results also suggest that wastewater treatment bioreactor design is pertinent, such that the imposition of feast-famine conditions (i.e., fill-and-draw bioreactors) nudge bacterial community assembly more towards stochastic mechanisms compared to the imposition of stringent nutrient limitation (i.e., continuous-flow bioreactors). Our research also validates the previously-used indirect methods (synchronous community dynamics and an application of a null-model) for characterizing the relative importance of deterministic versus stochastic mechanisms of community assembly.

Kinetic Analysis of Lidocaine Elimination by Pig Liver Cells Cultured in 3D Multi-Compartment Hollow Fiber Membrane Network Perfusion Bioreactors

Liver cells cultured in 3D bioreactors is an interesting option for temporary extracorporeal liver support in the treatment of acute liver failure and for animal models for preclinical drug screening. Bioreactor capacity to eliminate drugs is generally used for assessing cell metabolic competence in different bioreactors or to scale-up bioreactor design and performance for clinical or preclinical applications. However, drug adsorption and physical transport often disguise the intrinsic drug biotransformation kinetics and cell metabolic state. In this study, we characterized the intrinsic kinetics of lidocaine elimination and adsorption by porcine liver cells cultured in 3D four-compartment hollow fiber membrane network perfusion bioreactors. Models of lidocaine transport and biotransformation were used to extract intrinsic kinetic information from response to lidocaine bolus of bioreactor versus adhesion cultures. Different from 2D adhesion cultures, cells in the bioreactors are organized in liver-like aggregates. Adsorption on bioreactor constituents significantly affected lidocaine elimination and was effectively accounted for in kinetic analysis. Lidocaine elimination and cellular monoethylglicinexylidide biotransformation featured first-order kinetics with near-to-in vivo cell-specific capacity that was retained for times suitable for clinical assist and drug screening. Different from 2D cultures, cells in the 3D bioreactors challenged with lidocaine were exposed to close-to-physiological lidocaine and monoethylglicinexylidide concentration profiles. Kinetic analysis suggests bioreactor technology feasibility for preclinical drug screening and patient assist and that drug adsorption should be accounted for to assess cell state in different cultures and when laboratory bioreactor design and performance is scaled-up to clinical use or toxicological drug screening.