The human gut microbiome is a complex community of microorganisms. These microbes play an important role in host health by contributing essential compounds and acting as a barrier against pathogens. However, these communities and associated functions can be impacted by factors like disease and diet. In particular, microbial fermentation of dietary components like polysaccharides, proteins, and fats that reach the gut are being examined to better understand how these biopolymers are utilized and affect community structure. Thus, evaluating the accuracy of methods used to quantify specific macromolecules is crucial to gaining a precise understanding of how gut microbes hydrolyze those substrates. This study presents findings on the accuracy of the Megazyme RS kit (Rapid) modified for high performance liquid chromatography (HPLC) readings and the DC Protein Assay when performed on samples from complex gut media with potato starch treatments and bovine serum albumin (BSA) treatments. Overall, our data indicates that the megazyme RS kit needs further modification to detect expected starch content with the HPLC and that the DC Protein Assay is not suitable for specific protein analysis.

The biological carbon pump acts as part of the global carbon cycle through the photosynthetic fixation of inorganic carbon into dissolved and particulate organic carbon by phytoplankton. Previously, the biological carbon pump was attributed to large aggregates and zooplankton fecal pellets since their size and density results in faster sinking rates, efficiently exporting organic carbon to deeper depths in the ocean. However, recent studies have indicated that small cells, known as picoplankton, contribute significantly to the formation of sinking particles. The presence of exopolymeric substances (EPS), among them sticky transparent exopolymeric particles (TEP) and proteinaceous coomassie stainable particles (CSP), serve as influential factors of export flux and aggregation. The presence of heterotrophic bacteria can also affect aggregation and sinking velocity, as seen in previous studies, and is likely attributed to their EPS and TEP production. The staining and visualization of TEP and CSP allow for the qualitative determination of these types of EPS from bacteria isolated from sinking particles collected with particle interceptor traps at various depths in the Sargasso Sea. I study the presence of TEP and CSP in particle-associated bacteria. Cultures of picocyanobacteria, consisting of xenic Synechococcus and axenic Prochlorococcus, were used to establish positive and negative controls for stained isolate analysis. Marinobacter adhaerens served as a tertiary control for an axenic culture that stains positive for TEP. I chose six isolates of bacteria isolated from sinking particles to be stained and visualized to test for the secretion of TEP and CSP. Four of the isolates stained positive for both TEP and CSP, including Pseudoalteromonas sp., Erythrobacter sp., and Marinobacter sp., while one isolate, Micrococcus sp., stained positive only for TEP, and the last isolate, another Marinobacter sp., stained positive for only CSP. These results are important in understanding the role of plankton organisms in the formation of sinking particles.

The Combined Activated Sludge-Anaerobic Digestion Model (CASADM) quantifies the effects of recycling anaerobic-digester (AD) sludge on the performance of a hybrid activated sludge (AS)-AD system. The model includes nitrification, denitrification, hydrolysis, fermentation, methanogenesis, and production/utilization of soluble microbial products and extracellular polymeric substances (EPS). A CASADM example shows that, while effluent COD and N are not changed much by hybrid operation, the hybrid system gives increased methane production in the AD and decreased sludge wasting, both caused mainly by a negative actual solids retention time in the hybrid AD. Increased retention of biomass and EPS allows for more hydrolysis and conversion to methane in the hybrid AD. However, fermenters and methanogens survive in the AS, allowing significant methane production in the settler and thickener of both systems, and AD sludge recycle makes methane formation greater in the hybrid system.