Novel Eco-Friendly, Recycled Composites for Improved CA Road Surfaces

The continued use of structural plastics in consumer products, industry, and transportation represents a potential source for durable, long lasting, and recyclable roadways. Costs to dispose of reinforced plastics can be similar to procuring new asphalt with mechanical performance exceeding that of the traditional road surface. This project examines improved material development times by leveraging advanced computational material models based on validated experimental data. By testing traditional asphalt and select carbon and glass reinforced composites, both new and recycled, it is possible to develop a finite element simulation that can predict the material characteristics under a number of loads virtually, and with less lead time compared to experimental testing. From the tested specimens, composites show minimal strength degradation when recycled and used within the asphalt design envelopes considered, with an average of 49% less wear, two orders of magnitude higher compressive strength, and three orders for tensile strength. Predictive computational analysis using the validated material models developed for this investigation confirms the long-term durability

Function and Regulation of Vibrio campbellii Proteorhodopsin: Acquired Phototrophy in a Classical Organoheterotroph

Proteorhodopsins (PRs) are retinal-binding photoproteins that mediate light-driven proton translocation across prokaryotic cell membranes. Despite their abundance, wide distribution and contribution to the bioenergy budget of the marine photic zone, an understanding of PR function and physiological significance in situ has been hampered as the vast majority of PRs studied to date are from unculturable bacteria or culturable species that lack the tools for genetic manipulation. In this study, we describe the presence and function of a horizontally acquired PR and retinal biosynthesis gene cluster in the culturable and genetically tractable bioluminescent marine bacterium Vibrio campbellii. Pigmentation analysis, absorption spectroscopy and photoinduction assays using a heterologous over-expression system established the V. campbellii PR as a functional green light absorbing proton pump. In situ analyses comparing PR expression and function in wild type (WT) V. campbellii with an isogenic ΔpR deletion mutant revealed a marked absence of PR membrane localization, pigmentation and light-induced proton pumping in the ΔpR mutant. Comparative photoinduction assays demonstrated the distinct upregulation of pR expression in the presence of light and PR-mediated photophosphorylation in WT cells that resulted in the enhancement of cellular survival during respiratory stress. In addition, we demonstrate that the master regulator of adaptive stress response and stationary phase, RpoS1, positively regulates pR expression and PR holoprotein pigmentation. Taken together, the results demonstrate facultative phototrophy in a classical marine organoheterotrophic Vibrio species and provide a salient example of how this organism has exploited lateral gene transfer to further its adaptation to the photic zone

Marine genomics: at the interface of marine microbial ecology and biodiscovery

The composition and activities of microbes from diverse habitats have been the focus of intense research during the past decade with this research being spurred on largely by advances in molecular biology and genomic technologies. In recent years environmental microbiology has entered very firmly into the age of the ‘omics’– (meta)genomics, proteomics, metabolomics, transcriptomics – with probably others on the rise. Microbes are essential participants in all biogeochemical processes on our planet, and the practical applications of what we are learning from the use of molecular approaches has altered how we view biological systems. In addition, there is considerable potential to use information about uncultured microbes in biodiscovery research as microbes provide a rich source of discovery for novel genes, enzymes and metabolic pathways. This review explores the brief history of genomic and metagenomic approaches to study environmental microbial assemblages and describes some of the future challenges involved in broadening our approaches – leading to new insights for understanding environmental problems and enabling biodiscovery research

Assembling the Marine Metagenome, One Cell at a Time

The difficulty associated with the cultivation of most microorganisms and the complexity of natural microbial assemblages, such as marine plankton or human microbiome, hinder genome reconstruction of representative taxa using cultivation or metagenomic approaches. Here we used an alternative, single cell sequencing approach to obtain high-quality genome assemblies of two uncultured, numerically significant marine microorganisms. We employed fluorescence-activated cell sorting and multiple displacement amplification to obtain hundreds of micrograms of genomic DNA from individual, uncultured cells of two marine flavobacteria from the Gulf of Maine that were phylogenetically distant from existing cultured strains. Shotgun sequencing and genome finishing yielded 1.9 Mbp in 17 contigs and 1.5 Mbp in 21 contigs for the two flavobacteria, with estimated genome recoveries of about 91% and 78%, respectively. Only 0.24% of the assembling sequences were contaminants and were removed from further analysis using rigorous quality control. In contrast to all cultured strains of marine flavobacteria, the two single cell genomes were excellent Global Ocean Sampling (GOS) metagenome fragment recruiters, demonstrating their numerical significance in the ocean. The geographic distribution of GOS recruits along the Northwest Atlantic coast coincided with ocean surface currents. Metabolic reconstruction indicated diverse potential energy sources, including biopolymer degradation, proteorhodopsin photometabolism, and hydrogen oxidation. Compared to cultured relatives, the two uncultured flavobacteria have small genome sizes, few non-coding nucleotides, and few paralogous genes, suggesting adaptations to narrow ecological niches. These features may have contributed to the abundance of the two taxa in specific regions of the ocean, and may have hindered their cultivation. We demonstrate the power of single cell DNA sequencing to generate reference genomes of uncultured taxa from a complex microbial community of marine bacterioplankton. A combination of single cell genomics and metagenomics enabled us to analyze the genome content, metabolic adaptations, and biogeography of these taxa

A Simplified Methodology for Validating the Hyper-Viscoelastic (HVE) Dynamic Response

This thesis presents a mathematical modeling process for characterizing a hyperelastic material with viscous response under dynamic loading conditions. The model is designed with the advantage of performing only one compressive dynamic test in order to provide the requisite parameters to fully determine the hyper-viscoelastic response. This is achieved in both deformations and contact forces, using digital image correlation and force sensors. Experiments performed at strain rates ranging from 10–3–10 2 s–1 correlate with computational simulations at the same loading rates up to 80% compression. The validity of the fit and prediction is assessed using MATLAB along with ABAQUS finite element software. The results provided by this novel methodology, i.e. the mathematical model using non-homogeneous deformations and the subsequent dynamic experimental techniques, proves that this approach is a more effective alternative to the current standards used to characterize the mechanical response of hyperelastic, viscoelastic, and hyper-viscoelastic materials

Beyond the Iron Age: The ecological relevance of bioactive trace metals other than Fe and organic growth factors in aquatic systems

In the last three decades, research has extensively focused on the role of Fe and other mineral nutrients in regulating biological processes, ranging from the surface to the deep ocean. This has produced major breakthroughs in our understanding of the fundamental role of those bioactive elements on the carbon, nitrogen and sulfur cycles and ecosystem function. However, biological processes cannot be entirely sustained by that small set of chemical elements, and new scientific evidence suggests that trace metals other than Fe (e.g., Co, Mo and Ni) as well as essential organic growth factors (e.g., vitamins) may also be crucial in most aquatic systems

Response of Alteromonadaceae and Rhodobacteriaceae to glucose and phosphorus manipulation in marine mesocosms

Microbial successions were studied in experimental mesocosms of marine water in the presence of additional organic carbon (glucose), phosphorus (P) or both. P addition lead to pronounced blooms of phytoplankton and to significantly enhanced bacterial production. Characteristic succession patterns were observed for two phylogenetic groups of bacteria that both transiently formed > 50% of total cells. An initial bloom of bacteria affiliated to the Alteromonadaceae could not be assigned to any specific treatment and was interpreted as a response to the manipulations during mesocosm set-up. These bacteria rapidly declined with the appearance of heterotrophic nanoflagellates, suggesting a negative effect of selective grazing. The persistence of Alteromonadaceae in the microbial assemblages was significantly favored by the presence of additional glucose. During the second half of the experiment, bacteria affiliated to Rhodobacteriaceae formed a dominant component of the experimental assemblages in treatments with addition of P. The community contribution of Rhodobacteriaceae was significantly correlated with chlorophyll a concentrations only in the P-amended mesocosms (r(2) = 0.58). This was more pronounced in the absence of glucose (r(2) = 0.85). The phylogenetic and morphological diversity among Rhodobacteriaceae was high, and treatment-specific temporal successions of genotypes related to Rhodobacteriaceae were observed. We suggest that the observed succession patterns reflect different niche preferences: Alteromonadaceae rapidly responded to disturbance and profited from allochthonous glucose input, whereas Rhodobacteriaceae benefited from the phytoplankton bloom