Ecology and population structure of Vibrionaceae in the coastal ocean

Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the Massachusetts Institute of Technology and the Woods Hole oceanographic Institution February 2010Extensive genetic diversity has been discovered in the microbial world, yet mechanisms that shape and maintain this diversity remain poorly understood. This thesis investigates to what extent populations of the gamma-proteobacterial family, Vibrionaceae, are ecologically specialized by investigating the distribution across a wide range of environmental categories, such as marine invertebrates or particles in the water column. Additionally, it seeks to determine whether in situ population distributions directly result from a competitive advantage over other Vibrio populations. This was investigated by in vitro competition assays on mixtures of native, sterilized particles. Generalist populations were found to dominate the associations with marine invertebrates, consistent with a model of high migration dominated population assembly. A majority of populations occurred broadly within and among the different types of invertebrates sampled, with one population being a near perfect generalist with regard to seasons, host taxa and body regions. High variability across host individuals, consistent with a scenario of stochastic clonal expansion, was especially pronounced in crab and zooplankton samples. Specialization, demonstrated by specific and reproducible association with different particle types in the water column, is more common than specialization within invertebrate hosts. Co-existing Vibrio species show strong preferences for different types of particulate matter in the water column suggesting that competition for limited resources influences their evolution. While populations show different growth profiles on particle derived substrates, relative growth advantages of specialist populations in competition with other Vibrio populations on native particles may not be sufficient to explain observed environmental distributions. Instead, populations may gain an advantage on these particles by colonizing the living plant or zooplankton prior to death and degradation into particulate matter. In summary, although vibrios are known commensals of marine invertebrates, evidence suggests that population structure within animals is fairly weak compared to suspended particles in the water column. This highlights the importance of comparing multiple environmental categories and migration among them to investigate population structure and adaptation.I would like to thank the Linden Fellowship for their support during the Fall, 2003- Spring, 2004 and the Martin Family Society of Fellows for Sustainability for their support during the Fall 2005. Work in this thesis was also made possible by support from the following agencies: Woods Hole Center for Oceans and Human Health, which is funded jointly by the National Science Foundation (OCE-0430724) and the National Institute for Environmental Health Sciences (P50 ES012742); DOE-Chicago (DE-FG02- 08ER64516); The Gordon and Betty Moore Foundation (Letter Agreement 1108)

Bridge Girder Webs Subjected to Horizontal Loads

Common bridge construction practice in South Dakota involves suspending the deck finishing machine and the freshly poured deck overhang from temporary metal brackets. These brackets are attached to the webs of the exterior plate girders. When loaded during construction, the brackets transmit to the web a vertical shearing force plus a couple. Since the webs are not designed for carrying horizontal loads, this type of loading could overstress the webs and appreciably lower their ultimate resistance to buckling. With specifications now permitting large depth to thickness ratios, deflections as well as stresses may be excessive. Rotation of the brackets, caused by deflection of the webs, lowers the paving machine and could result in undesirable thinning of the slab. To reduce stresses and deflections, contractors are now required to place the brackets within six inches of a lateral stiffener. However, because stiffener spacing varies from bridge to bridge, standardization of formwork becomes impossible and the resulting bracket spacing may not always be the most economical. By developing a bracket which could be used without regard to stiffener spacing, construction time and cost could be reduced

Polyphyly of non-bioluminescent Vibrio fischeri sharing a lux-locus deletion

available in PMC 2013 May 16This study reports the first description and molecular characterization of naturally occurring, non-bioluminescent strains of Vibrio fischeri. These ‘dark’V. fischeri strains remained non-bioluminescent even after treatment with both autoinducer and aldehyde, substrate additions that typically maximize light production in dim strains of luminous bacteria. Surprisingly, the entire lux locus (eight genes) was absent in over 97% of these dark V. fischeri strains. Although these strains were all collected from a Massachusetts (USA) estuary in 2007, phylogenetic reconstructions allowed us to reject the hypothesis that these newly described non-bioluminescent strains exhibit monophyly within the V. fischeri clade. These dark strains exhibited a competitive disadvantage against native bioluminescent strains when colonizing the light organ of the model V. fischeri host, the Hawaiian bobtail squid Euprymna scolopes. Significantly, we believe that the data collected in this study may suggest the first observation of a functional, parallel locus-deletion event among independent lineages of a non-pathogenic bacterial species.National Institutes of Health (U.S.) (NIH Molecular Biosciences (5T32GM007215-35))National Institutes of Health (U.S.) (NIH Microbes in Health and Disease, training grant (2T32AI055397-07))Gordon and Betty Moore FoundationBroad Institute of MIT and Harvard (SPARC programme)National Science Foundation (U.S.) (NSF IOS 0841507)National Institutes of Health (U.S.) (NIH R01 RR12294)National Science Foundation (U.S.) (NSF Microbial Systems in the Biosphere programme)Woods Hole Center for Oceans & Human Healt

Cirrhosis-induced defects in innate pulmonary defenses against Streptococcus pneumoniae

<p>Abstract</p> <p>Background</p> <p>The risk of mortality from pneumonia caused by <it>Streptococcus pneumoniae </it>is increased in patients with cirrhosis. However, the specific pneumococcal virulence factors and host immune defects responsible for this finding have not been clearly established. This study used a cirrhotic rat model of pneumococcal pneumonia to identify defect(s) in innate pulmonary defenses in the cirrhotic host and to determine the impact of the pneumococcal toxin pneumolysin on these defenses in the setting of severe cirrhosis.</p> <p>Results</p> <p>No cirrhosis-associated defects in mucociliary clearance of pneumococci were found in these studies, but early intrapulmonary killing of the organisms before the arrival of neutrophils was significantly impaired. This defect was exacerbated by pneumolysin production in cirrhotic but not in control rats. Neutrophil-mediated killing of a particularly virulent type 3 pneumococcal strain also was significantly diminished within the lungs of cirrhotic rats with ascites. Levels of lysozyme and complement component C3 were both significantly reduced in bronchoalveolar lavage fluid from cirrhotic rats. Finally, complement deposition was reduced on the surface of pneumococci recovered from the lungs of cirrhotic rats in comparison to organisms recovered from the lungs of control animals.</p> <p>Conclusion</p> <p>Increased mortality from pneumococcal pneumonia in this cirrhotic host is related to defects in both early pre-neutrophil- and later neutrophil-mediated pulmonary killing of the organisms. The fact that pneumolysin production impaired pre-neutrophil-mediated pneumococcal killing in cirrhotic but not control rats suggests that pneumolysin may be particularly detrimental to this defense mechanism in the severely cirrhotic host. The decrease in neutrophil-mediated killing of pneumococci within the lungs of the cirrhotic host is related to insufficient deposition of host proteins such as complement C3 on their surfaces. Pneumolysin likely plays a role in complement consumption within the lungs. Our studies, however, were unable to determine whether pneumolysin more negatively impacted this defense mechanism in cirrhotic than in control rats. These findings contribute to our understanding of the defects in innate pulmonary defenses that lead to increased mortality from pneumococcal pneumonia in the severely cirrhotic host. They also suggest that pneumolysin may be a particularly potent pneumococcal virulence factor in the setting of cirrhosis.</p

High resolution time series reveals cohesive but short-lived communities in coastal plankton

© The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Nature Communications 9 (2018): 266, doi:10.1038/s41467-017-02571-4.Because microbial plankton in the ocean comprise diverse bacteria, algae, and protists that are subject to environmental forcing on multiple spatial and temporal scales, a fundamental open question is to what extent these organisms form ecologically cohesive communities. Here we show that although all taxa undergo large, near daily fluctuations in abundance, microbial plankton are organized into clearly defined communities whose turnover is rapid and sharp. We analyze a time series of 93 consecutive days of coastal plankton using a technique that allows inference of communities as modular units of interacting taxa by determining positive and negative correlations at different temporal frequencies. This approach shows both coordinated population expansions that demarcate community boundaries and high frequency of positive and negative associations among populations within communities. Our analysis thus highlights that the environmental variability of the coastal ocean is mirrored in sharp transitions of defined but ephemeral communities of organisms.This work was supported by grants from the U.S. National Science Foundation (OCE-1441943) to M.F.P. and the U.S. Department of Energy (DE-SC0008743) to M.F.P. and E.J.A. A.M.M.-P. was partially supported by the Ramon Areces foundation through a postdoctoral fellowship. D.J.M. was supported by the U.S. National Science Foundation (OCE-1314642) and National Institute of Environmental Health Sciences (1P01ES021923-01) through the Woods Hole Center for Oceans and Human Health

Natural Bacterial Communities Serve as Quantitative Geochemical Biosensors

Biological sensors can be engineered to measure a wide range of environmental conditions. Here we show that statistical analysis of DNA from natural microbial communities can be used to accurately identify environmental contaminants, including uranium and nitrate at a nuclear waste site. In addition to contamination, sequence data from the 16S rRNA gene alone can quantitatively predict a rich catalogue of 26 geochemical features collected from 93 wells with highly differing geochemistry characteristics. We extend this approach to identify sites contaminated with hydrocarbons from the Deepwater Horizon oil spill, finding that altered bacterial communities encode a memory of prior contamination, even after the contaminants themselves have been fully degraded. We show that the bacterial strains that are most useful for detecting oil and uranium are known to interact with these substrates, indicating that this statistical approach uncovers ecologically meaningful interactions consistent with previous experimental observations. Future efforts should focus on evaluating the geographical generalizability of these associations. Taken as a whole, these results indicate that ubiquitous, natural bacterial communities can be used as in situ environmental sensors that respond to and capture perturbations caused by human impacts. These in situ biosensors rely on environmental selection rather than directed engineering, and so this approach could be rapidly deployed and scaled as sequencing technology continues to become faster, simpler, and less expensive

Population Genomics of Early Events in the Ecological Differentiation of Bacteria

Genetic exchange is common among bacteria, but its effect on population diversity during ecological differentiation remains controversial. A fundamental question is whether advantageous mutations lead to selection of clonal genomes or, as in sexual eukaryotes, sweep through populations on their own. Here, we show that in two recently diverged populations of ocean bacteria, ecological differentiation has occurred akin to a sexual mechanism: A few genome regions have swept through subpopulations in a habitat-specific manner, accompanied by gradual separation of gene pools as evidenced by increased habitat specificity of the most recent recombinations. These findings reconcile previous, seemingly contradictory empirical observations of the genetic structure of bacterial populations and point to a more unified process of differentiation in bacteria and sexual eukaryotes than previously thought.National Science Foundation (U.S.) (Grant DEB-0918333)Woods Hole Center for Oceans & Human HealthGordon and Betty Moore FoundationUnited States. Dept. of Energy. Genomes To Lif