RESUMO
The use of online mass spectrometry for detecting volatile organic compounds (VOCs) has proven to be a powerful technique, allowing for real-time analysis of many chemical and biochemical processes. Unfortunately, online mass spectrometry has had limited application due to high instrument costs and limited availability. Here, we detail the design, construction, and performance characteristics of a custom ion-molecule reactor retrofitted to a commonly used single quadrupole mass spectrometer to operate as an online chemical ionization mass spectrometer (CIMS). This low-cost modified CIMS is capable of limits of detection below 10 parts per trillion for select VOCs including dimethyl sulfide, dimethylamine, and trimethylamine.
RESUMO
The accumulation of microplastics in various ecosystems has now been well documented and recent evidence suggests detrimental effects on various biological processes due to this pollution. Accumulation of microplastics in the natural environment is ultimately due to the chemical nature of widely used petroleum-based plastic polymers, which typically are inaccessible to biological processing. One way to mitigate this crisis is adoption of plastics that biodegrade if released into natural environments. In this work, we generated microplastic particles from a bio-based, biodegradable thermoplastic polyurethane (TPU-FC1) and demonstrated their rapid biodegradation via direct visualization and respirometry. Furthermore, we isolated multiple bacterial strains capable of using TPU-FC1 as a sole carbon source and characterized their depolymerization products. To visualize biodegradation of TPU materials as real-world products, we generated TPU-coated cotton fabric and an injection molded phone case and documented biodegradation by direct visualization and scanning electron microscopy (SEM), both of which indicated clear structural degradation of these materials and significant biofilm formation.
Assuntos
Plásticos , Poliuretanos , Plásticos/química , Poliuretanos/química , Microplásticos , Ecossistema , Biodegradação AmbientalRESUMO
Cyanobacteria are attracting increasing attention as a photosynthetic chassis organism for diverse biochemical production, however, photoautotrophic production remains inefficient. Photomixotrophy, a method where sugar is used to supplement baseline autotrophic metabolism in photosynthetic hosts, is becoming increasingly popular for enhancing sustainable bioproduction with multiple input energy streams. In this study, the commercially relevant diacid, succinate, was produced photomixotrophically. Succinate is an important industrial chemical that can be used for the production of a wide array of products, from pharmaceuticals to biopolymers. In this system, the substrate, glucose, is transported by a proton symporter and the product, succinate, is hypothesized to be transported by another proton symporter, but in the opposite direction. Thus, low pH is required for the import of glucose and high pH is required for the export of succinate. Succinate production was initiated in a pH 7 medium containing bicarbonate. Glucose was efficiently imported at around neutral pH. Utilization of bicarbonate by CO2 fixation raised the pH of the medium. As succinate, a diacid, was produced, the pH of the medium dropped. By repeating this cycle with additional pH adjustment, those contradictory requirements for transport were overcome. pH affects a variety of biological factors and by cycling from high pH to neutral pH processes such as CO2 fixation rates and CO2 solubility can vary. In this study the engineered strains produced succinate during fluctuating pH conditions, achieving a titer of 5.0 g L-1 after 10 days under shake flask conditions. These results demonstrate the potential for photomixotrophic production as a viable option for the large-scale production of succinate.
Assuntos
Ácido Succínico , Simportadores , Ácido Succínico/metabolismo , Dióxido de Carbono/metabolismo , Prótons , Bicarbonatos/metabolismo , Engenharia Metabólica/métodos , Succinatos/metabolismo , Glucose/metabolismo , Concentração de Íons de HidrogênioRESUMO
Accumulation of plastics in the Earth's oceans is causing widespread disruption to marine ecosystems. To help mitigate the environmental burden caused by non-degradable plastics, we have previously developed a commercially relevant polyurethane (PU) foam derived from renewable biological materials that can be depolymerized into its constituent monomers and consumed by microorganisms in soil or compost. Here we demonstrate that these same PU foams can be biodegraded by marine microorganisms in the ocean and by isolated marine microorganisms in an ex situ seawater environment. Using Fourier-transform infrared (FTIR) spectroscopy, we tracked molecular changes imparted by microbial breakdown of the PU polymers; and utilized scanning electron microscopy (SEM) to demonstrate the loss of physical structure associated with colonization of microorganisms on the PU foams. We subsequently enriched, isolated, and identified individual microorganisms, from six marine sites around San Diego, CA, that are capable of depolymerizing, metabolizing, and accumulating biomass using these PU foams as a sole carbon source. Analysis using SEM, FTIR, and gas chromatography-mass spectrometry (GCMS) confirmed that these microorganisms depolymerized the PU into its constitutive diols, diacids, and other PU fragments. SEM and FTIR results from isolated organismal biodegradation experiments exactly matched those from ex situ and ocean biodegradation samples, suggesting that these PU foam would undergo biodegradation in a natural ocean environment by enzymatic depolymerization of the PU foams and eventual uptake of the degradation products into biomass by marine microorganisms, should these foams unintentionally end up in the marine environment, as many plastics do.
Assuntos
Ecossistema , Poliuretanos , Biodegradação Ambiental , Carbono , Plásticos , Poliuretanos/química , SoloRESUMO
Algae cultivation in open raceway ponds is considered the most economical method for photosynthetically producing biomass for biofuels, chemical feedstocks, and other high-value products. One of the primary challenges for open ponds is diminished biomass yields due to attack by grazers, competitors, and infectious organisms. Higher-frequency observations are needed for detection of grazer infections, which can rapidly reduce biomass levels. In this study, real-time measurements were performed using chemical ionization mass spectrometry (CIMS) to monitor the impact of grazer infections on cyanobacterial cultures. Numerous volatile gases were produced during healthy growth periods from freshwater Synechococcus elongatus Pasteur Culture Collection (PCC) 7942, with 6-methyl-5-hepten-2-one serving as a unique metabolic indicator of exponential growth. Following the introduction of a Tetrahymena ciliate grazer, the concentrations of multiple volatile species were observed to change after a latent period as short as 18 h. Nitrogenous gases, including ammonia and pyrroline, were found to be reliable indicators of grazing. Detection of grazing by CIMS showed indicators of infections much sooner than traditional methods, microscopy, and continuous fluorescence, which did not detect changes until 37 to 76 h after CIMS detection. CIMS analysis of gases produced by PCC 7942 further shows a complex temporal array of biomass-dependent volatile gas production, which demonstrates the potential for using volatile gas analysis as a diagnostic for grazer infections. Overall, these results show promise for the use of continuous volatile metabolite monitoring for the detection of grazing in algal monocultures, potentially reducing current grazing-induced biomass losses, which could save hundreds of millions of dollars.
Assuntos
Biocombustíveis/análise , Cianobactérias/química , Gases/química , Biomassa , LagoasRESUMO
Environmental conservation initiatives, including marine protected areas (MPAs), have proliferated in recent decades. Designed to conserve marine biodiversity, many MPAs also seek to foster sustainable development. As is the case for many other environmental policies and programs, the impacts of MPAs are poorly understood. Social-ecological systems, impact evaluation, and common-pool resource governance are three complementary scientific frameworks for documenting and explaining the ecological and social impacts of conservation interventions. We review key components of these three frameworks and their implications for the study of conservation policy, program, and project outcomes. Using MPAs as an illustrative example, we then draw upon these three frameworks to describe an integrated approach for rigorous empirical documentation and causal explanation of conservation impacts. This integrated three-framework approach for impact evaluation of governance in social-ecological systems (3FIGS) accounts for alternative explanations, builds upon and advances social theory, and provides novel policy insights in ways that no single approach affords. Despite the inherent complexity of social-ecological systems and the difficulty of causal inference, the 3FIGS approach can dramatically advance our understanding of, and the evidentiary basis for, effective MPAs and other conservation initiatives.
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Biodiversidade , Conservação dos Recursos Naturais/métodos , Ecossistema , Biologia Marinha/métodos , Animais , Organismos Aquáticos/classificação , Organismos Aquáticos/fisiologia , Conservação dos Recursos Naturais/economia , Conservação dos Recursos Naturais/legislação & jurisprudência , Política Ambiental/economia , Política Ambiental/legislação & jurisprudência , Humanos , Biologia Marinha/economia , Biologia Marinha/legislação & jurisprudência , Modelos Teóricos , Fatores SocioeconômicosRESUMO
High-resolution melt (HRM) analysis of the VNTR region of the human D1S80 locus, a 16-bp repeat minisatellite from approximately 400 to over 700 bp in length, was investigated. A Qiagen Rotor-Gene Q using the Type-it PCR HRM kit was used to acquire HRM curves for 14 single, and 16 biallelic, dsDNA samples. The HRM analysis was applicable over a range of DNA concentrations; however the characteristics of the melt curve did depend on the forward and reverse primer ratio. Despite the large amplicon size and the similarities of the repeat sequences, it was possible to discriminate different genotypes. Heterozygotes were clearly different from the homozygous variants and even small differences in the repeat sequence could be differentiated. However, the melt analysis requires a high-resolution system with temperature resolution of 0.02°C or better in order to sort out differences in these large amplicons of near identical GC content (in this case 56%). HRM analysis of amplicons with large repeat sequences can be used as a means of comparing DNA fragments. Examination of multiple sequences can be used to differentiate DNA samples and demonstrate the potential of HRM analysis as a rapid and inexpensive prescreening technique in forensic applications.
Assuntos
DNA/análise , Genética Forense/métodos , Repetições Minissatélites/genética , Desnaturação de Ácido Nucleico/genética , Análise de Sequência de DNA/métodos , Composição de Bases , DNA/genética , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo ÚnicoRESUMO
Over 1.3 billion people live on tropical coasts, primarily in developing countries. Many depend on adjacent coastal seas for food, and livelihoods. We show how trends in demography and in several local and global anthropogenic stressors are progressively degrading capacity of coastal waters to sustain these people. Far more effective approaches to environmental management are needed if the loss in provision of ecosystem goods and services is to be stemmed. We propose expanded use of marine spatial planning as a framework for more effective, pragmatic management based on ocean zones to accommodate conflicting uses. This would force the holistic, regional-scale reconciliation of food security, livelihoods, and conservation that is needed. Transforming how countries manage coastal resources will require major change in policy and politics, implemented with sufficient flexibility to accommodate societal variations. Achieving this change is a major challenge - one that affects the lives of one fifth of humanity.
Assuntos
Mudança Climática , Conservação dos Recursos Naturais , Ecossistema , Animais , Antozoários , Simulação por Computador , Demografia , Ecologia , Monitoramento Ambiental/métodos , Pesqueiros , Geografia , Humanos , Oceanos e MaresRESUMO
In the ocean, breaking waves generate air bubbles which burst at the surface and eject sea spray aerosol (SSA), consisting of sea salt, biogenic organic species, and primary biological aerosol particles (PBAP). Our overall understanding of atmospheric biological particles of marine origin remains poor. Here, we perform a control experiment, using an aerosol time-of-flight mass spectrometer to measure the mass spectral signatures of individual particles generated by bubbling a salt solution before and after addition of heterotrophic marine bacteria. Upon addition of bacteria, an immediate increase occurs in the fraction of individual particle mass spectra containing magnesium, organic nitrogen, and phosphate marker ions. These biological signatures are consistent with 21% of the supermicrometer SSA particles generated in a previous study using breaking waves in an ocean-atmosphere wave channel. Interestingly, the wave flume mass spectral signatures also contain metal ions including silver, iron, and chromium. The nascent SSA bioparticles produced in the wave channel are hypothesized to be as follows: (1) whole or fragmented bacterial cells which bioaccumulated metals and/or (2) bacteria-derived colloids or biofilms which adhered to the metals. This study highlights the potential for transition metals, in combination with specific biomarkers, to serve as unique indicators for the presence of marine PBAP, especially in metal-impacted coastal regions.
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Aerossóis/análise , Organismos Aquáticos/química , Oceanos e Mares , Material Particulado/análise , Elementos de Transição/análise , Movimentos da Água , Espectrometria de Massas , Microscopia Eletrônica de Transmissão , Tamanho da PartículaRESUMO
Early efforts to apply the concept of fisheries co-management in Southeast Asia focused primarily on building the effectiveness of local management institutions and advocating the merits of the approach so that it would be applied in new sites, while gradually learning and adapting to a range of obstacles in practice. Today, with co-management widely embraced by the research community and adopted as policy by an increasing number of governments, a second-generation perspective has emerged. This perspective is distinguished by efforts to navigate and influence change in the broader institutional and governance context: (a) a more sophisticated appreciation of politics, power relations, and the role of the state, (b) efforts to manage resource competition beyond the fisheries sector, (c) building institutions for adaptation and learning, and (d) recognizing divergent values and goals influencing fisheries management. This paper traces the evolution of this second-generation perspective, noting how it has built on learning from early practice and how it has been cross-fertilized by theoretical innovations in related fields, notably resilience thinking and political ecology. We illustrate this evolution through analysis of experience in the Philippines, with a relatively long experience of learning and adaptation in fisheries co-management practice, and Vietnam, where fisheries co-management policies have been embraced more recently. Characterizing the second-generation perspective helps identify points of convergence in the research and policy community about what needs attention, providing a basis for more systematic cross-country and cross-regional learning.