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DOE OSTI2020

In Vivo Molecular Insights into Syntrophic Geobacter Aggregates

Direct interspecies electron transfer (DIET) has been considered as a novel and highly efficient strategy in both natural anaerobic environments and artificial microbial fuel cells. A syntrophic model consisting of Geobacter metallireducens and Geobacter sulfurreducens was studied in this work. We conducted in vivo molecular mapping of the outer surface of the syntrophic community as the interface of nutrients and energy exchange. System for Analysis at the Liquid Vacuum Interface combined with time-of-flight secondary ion mass spectrometry was employed to capture the molecular distribution of syntrophic Geobacter communities in the living and hydrated state. Principal component analysis with selected peaks revealed that syntrophic Geobacter aggregates were well differentiated from other control samples, including syntrophic planktonic cells, pure cultured planktonic cells, and single population biofilms. Our in vivo imaging indicated that a unique molecular surface was formed. Specifically, aromatic amino acids, phosphatidylethanolamine components, and large water clusters were identified as key components that favored the DIET of syntrophic Geobacter aggregates. Moreover, the molecular changes in depths of the Geobacter aggregates were captured using dynamic depth profiling. Our findings shed new light on the interface components supporting electron transfer in syntrophic communities based on in vivo molecular imaging.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2023

Cooperative microbial interactions drive spatial segregation in porous environments

The role of microbial interactions and the underlying mechanisms that shape complex biofilm communities are poorly understood. Here we employ a microfluidic chip to represent porous subsurface environments and show that cooperative microbial interactions between free-living and biofilm-forming bacteria trigger active spatial segregation to promote their respective dominance in segregated microhabitats. During initial colonization, free-living and biofilm-forming microbes are segregated from the mixed planktonic inoculum to occupy the ambient fluid and grain surface. Contrary to spatial exclusion through competition, the active spatial segregation is induced by cooperative interactions which improves the fitness of both biofilm and planktonic populations. We further show that free-living Arthrobacter induces the surface colonization by scavenging the biofilm inhibitor, D-amino acids and receives benefits from the public goods secreted by the biofilm-forming strains. Collectively, our results reveal how cooperative microbial interactions may contribute to microbial coexistence in segregated microhabitats and drive subsurface biofilm community succession.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Origin of biogeographically distinct ecotypes during laboratory evolution

Resource partitioning is central to the incredible productivity of microbial communities, including gigatons in annual methane emissions through syntrophic interactions. Previous work revealed how a sulfate reducer (Desulfovibrio vulgaris, Dv) and a methanogen (Methanococcus maripaludis, Mm) underwent evolutionary diversification in a planktonic context, improving stability, cooperativity, and productivity within 300-1000 generations. Here, we show that mutations in just 15 Dv and 7 Mm genes within a minimal assemblage of this evolved community gave rise to co-existing ecotypes that were spatially enriched within a few days of culturing in a fluidized bed reactor. The spatially segregated communities partitioned resources in the simulated subsurface environment, with greater lactate utilization by attached Dv but partial utilization of resulting H 2 by low affinity hydrogenases of Mm in the same phase. The unutilized H 2 was scavenged by high affinity hydrogenases of planktonic Mm, producing copious amounts of methane. Our findings show how a few mutations can drive resource partitioning amongst niche-differentiated ecotypes, whose interplay synergistically improves productivity of the entire mutualistic community.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Changes in environmental and engineered conditions alter the plasma membrane lipidome of fractured shale bacteria

ABSTRACT Microorganisms that persist in fractured shale reservoirs cause several problems including secreting foul gases and forming biofilms. Current biocontrol measures often fail due to limited knowledge of their in situ activities. The plasma membrane protects the cell, mediates many of its critical functions, and responds to intracellular cues and ecological perturbations through physicochemical modifications. As such, it provides valuable insight into the physiological adaptation of microorganisms in disturbed environmental systems. Here, we (i) demonstrate how changes in salinity and hydraulic retention time (HRT) influence the plasma membrane intact polar lipid (IPL) chemistry of model bacterium, Halanaerobium congolense WG10, and mixed microbial consortia enriched from shale-produced fluids and (ii) elucidate adjustments in membrane IPL chemistry during biofilm growth relative to planktonic cells. We incubated H. congolense WG10 in chemostats under three salinities (7%, 13%, and 20% NaCl), operated under three HRTs (19.2, 24, and 48 h), and in drip flow biofilm reactors under the same salinity gradients. Also, mixed microbial consortia in produced fluids were enriched in triplicate chemostat vessels under three HRTs (19.2, 24, and 72 h) and biofilm reactors. Lipids were analyzed by ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Our results show that phosphatidylglycerols, cardiolipins, and phosphatidylethanolamines were predominantly enriched in planktonic H. congolense WG10 cells grown at hypersalinity (20%) compared to optimum (13%). In addition, several zwitterionic phosphatidylcholines and phosphatidylethanolamines were higher in abundance during biofilm growth. These observations suggest that microbial adaptation and biofilm formation in fractured shale are enabled by strategic plasma membrane IPL chemistry adjustments. IMPORTANCE Microorganisms inadvertently introduced into the shale reservoir during fracturing face multiple stressors including brine-level salinities and starvation. However, some anaerobic halotolerant bacteria adapt and persist for long periods of time. They produce hydrogen sulfide, which sours the reservoir and corrodes engineering infrastructure. In addition, they form biofilms on rock matrices, which decrease shale permeability and clog fracture networks. These reduce well productivity and increase extraction costs. Under stress, microbes remodel their plasma membrane to optimize its roles in protection and mediating cellular processes such as signaling, transport, and energy metabolism. Hence, by observing changes in the membrane lipidome of model shale bacteria, Halanaerobium congolense WG10, and mixed consortia enriched from produced fluids under varying subsurface conditions and growth modes, we provide insight that advances our knowledge of the fractured shale biosystem. We also offer data-driven recommendations for improving biocontrol efficacy and the efficiency of energy recovery from unconventional formations.

03 NATURAL GAS↗

DOE OSTI2026

Characterization of Pseudomonas aeruginosa and Acinetobacter calcoaceticus-baumannii complex traumatic wound isolates

Healing of traumatic wounds is complicated by infecting pathogens, with Pseudomonas aeruginosa and members of the Acinetobacter calcoaceticus-baumannii complex among the most common infectious agents. However, a full understanding of genotypic and phenotypic differences between circulating wound isolates is lacking. To address this, traumatic wounds were sampled at Emory University Hospital, and 24 isolates were characterized; we focused on P. aeruginosa and Acinetobacter due to their prevalence and tendency for antibiotic resistance. Even though these species are renowned for antibiotic resistance, only two of the tested isolates could be classified as multidrug resistant. Whole-genome sequencing and analysis revealed that isolates from different patients were genetically distinct; however, longitudinal isolates from the same patient were closely related and appeared to represent chronic colonization by the same strain. Phylogenetic analysis revealed that laboratory strains (PAO1 and AB5075) that were isolated decades previously and from different locations grouped closely to subsets of the recent wound isolates. Given the importance of biofilm formation in infection, the ability of the isolates to form biofilms was assessed; all isolates formed biofilms but showed temporal and magnitude differences. Analysis of a subset of isolates revealed that planktonic P. aeruginosa was resistant to human serum-mediated killing. While the same was true for the majority of Acinetobacter isolates, one showed planktonic sensitivity that was abrogated when grown in a biofilm. Taken together, these data reveal genotypic and phenotypic differences in circulating isolates of P. aeruginosa and A. baumannii.

Acinetobacter↗

NASA NTRS1973

Biomass in the upwelling areas along the northwest coast of Africa as viewed with ERTS-1

Light penetration in water is affected by plankton, algae, and dissolved and suspended matter. As a consequence, the composition of backscattered light from below the air-sea interface is determined by the nature of the constituents in the water column. In contrast to the absorption spectrum of chemically pure chlorophyll in solution, algae suspensions absorb and scatter light more uniformly throughout the visible part of the electromagnetic spectrum. Because of their spectral absorption and scattering properties plankton concentration can be estimated by measuring the spectral backscattered radiance over water. Experiments using this approach were performed in upwelling regions along the northwest coast of Africa.

Szekielda, K.↗

NASA NTRS1973

Water quality parameters of harbors of Charlotte Amalie, St. Thomas, Virgin Islands: Acquisition of in situ water data, intercorrelation of selected water parameters, and initial correlation of these in situ biological, chemical and physical data with ERTS-1 bulk CCT MSS band 5 data

The author has identified the following significant results. Remote sensing by the ERTS-1 satellite was compared with selected water quality parameters including pH, salinity, conductivity, dissolved oxygen, water depth, water temperature, turbidity, plankton concentration, current variables, chlorophylla, total carotenoids, and species diversity of the benthic community. Strong correlation between turbidity and MSS-sensed radiance was recorded and less strong correlations between the two plankton pigments and radiance. Turbidity and benthic species diversity were highly correlated furnishing an inferential tie between an easily sensed water quality variable and a sensitive indicator of average water quality conditions.

Coulbourn, W. C.↗

NASA NTRS1975

Reconnaissance of marine resources

A test area along the NW Coast of Africa was used during the Skylab mission to study the distribution of temperature and plankton. The S190B Earth Terrain Camera with a spectral film response of 0.4-0.7 micrometers allowed qualitative estimates of the distribution patterns of suspended material. Differentiation between inorganic particles and phytoplankton could be made by comparing the green band and the red band of the S190A Camera System. The pictorial display of data obtained from the S191 scanning radiometer in the 10-11 micrometer atmospheric window allowed a detailed interpretation of the temperature distribution in the area where cold upwelled water reaches the euphotic zone. The comparison between infrared data and the imageries taken simultaneously indicated the origin of the cold water as well as the pathway within the Canary current. A fish survey carried out almost simultaneously in the area, by echosounding, showed high correlation between the position of good fishing grounds and the distribution of plankton as detected by remote sensing detectors on Skylab.

Szekielda, K.-H.↗

NASA NTRS1988

Cretaceous-Tertiary boundary in the Antarctic: Climatic cooling precedes biotic crisis

Stable isotopic investigations were conducted on calcareous microfossils across two deep sea Cretaceous-Tertiary boundary sequences on Maud Rise, Weddell Sea, Antarctica. The boundary is taken at the level of massive extinctions in calcareous planktonic microfossils, and coincides with a sharp lithologic change from pure calcareous ooze to calcareous ooze with a large volcanic clay component. The uppermost Maestrichtian is marked by a long-term decrease in delta value of 0 to 18 which spans most of the lower and middle A. mayaroensis Zone and represents a warming trend which culminated in surface water temperatures of about 16 C. At approximately 3 meters below the K-T boundary this warming trend terminates abruptly and benthic and planktonic isotopic records exhibit a rapid increase in delta value of 0 to 18 that continues up to the K-T boundary. The trend towards cooler surface water temperatures stops abruptly at the K-T boundary and delta value of 0 to 18 values remain relatively stable through the Paleocene. Comparison of the Antarctic sequence with the previously documented deep sea records in the South Atlantic reveal shifts of similar magnitude in the latest Maestrichtian. It is indicated that the Southern Ocean underwent the most significant, and apparently permanent, climatic change. The latest Cretaceous oxygen isotopic shift recorded at Maud Rise and other deep sea sites is similar in magnitude to large positive delta valve of 0 to 18 shifts in the middle Eocene, at the Eocene/Oligocene boundary and in the middle Miocene that marked large scale climatic transitions which ultimately lead to cryospheric development of the Antarctic. The climatic shift at the end of the Cretaceous represents one of the most significant climatic transitions recorded in the latest Phanerozoic and had a profound effect on global climate as well as oceanic circulation.

Stott, Lowell D.↗

NASA NTRS1988

The Cretaceous-Tertiary boundary marine extinction and global primary productivity collapse

The extinction of marine phyto-and zoo-plankton across the K-T boundary has been well documented. Such an event may have resulted in decreased photosynthetic fixation of carbon in surface waters and a collapse of the food chain in the marine biosphere. Because the vertical and horizontal distribution of the carbon isotopic composition of total dissolved carton (TDC) in the modern ocean is controlled by the transfer of organic carbon from the surface to deep reservoirs, it follows that a major disruption of the marine biosphere would have had a major effect on the distribution of carbon isotopes in the ocean. Negative carbon isotope excursions have been identified at many marine K-T boundary sequences worldwide and are interpreted as a signal of decreased oceanic primary productivity. However, the magnitude, duration and consequences of this productivity crisis have been poorly constrained. On the basis of planktonic and benthic calcareous microfossil carbon isotope and other geochemical data from DSDP Site 577 located on the Shatsky Rise in the north-central Pacific, as well as other sites, researchers have been able to provide a reasonable estimate of the duration and magnitude of this event.

Zachos, J. C.↗

NASA NTRS1996

Volume 35, AMT-1 Cruise Report and Preliminary Results

This report documents the scientific activities on board the Royal Research Ship (RRS) 'James Clark Ross' during the irst Atlantic Meridional Transect (AMT-1), 21 September to 24 October 1995. The ship sailed from Grimsby (England) for Montevideo (Uruguay) and then continued on to Stanley (Falkland Islands). The primary objective of the AMT program is to investigate basic biological processes in the open Atlantic Ocean over very broad spatial scales. For AMT-1, the meridional range covered was approximately 50 deg N to 50 deg S or nearly 8,000 nmi. The measurements to be taken during the AMT cruises are fundamental for the calibration, validation, and continuing understanding of remotely sensed observations of biological oceanography. They are also important for understanding plankton community structure over latitudinal scales and the role of the world ocean in global carbon cycles. During AMT-1 a variety of instruments were used to map the physical, chemical, and biological structure of the upper 200 m of the water column. Ocean color measurements were made using state-of-the-art sensors, whose calibration was traceable to the highest international standards. New advances in fluorometry were used to measure photosynthetic activity, which was then used to further interpret primary productivity. A unique set of samples and data were collected for the planktonic assemblages that vary throughout the range of the transect. These data will yield new interpretations on community composition and their role in carbon cycling. While the various provinces of the Atlantic Ocean were being crossed, the partial pressure of CO2 was related to biological productivity. This comparison revealed the areas of drawdown of atmospheric CO2 and how these areas relate to the surrounding biological productivity. These data, plus the measurements of light attenuation and phytoplankton optical properties, will be used as a primary input for basin-scale biological productivity models to help develop ecosystem dynamics models which will be important for improving the forecasting abilities of modelers. The AMT program is also attempting to meet the needs of international agencies in their implementation of Sensor Intercomparison and Merger for Biological and Interdisciplinary Ocean Studies (SIMBIOS), a program to develop a methodology and operational capability to combine data products from the various ocean color satellite missions.

Hooker, Stanford B.↗

NASA NTRS2001

Models of Small-Scale Patchiness

Patchiness is perhaps the most salient characteristic of plankton populations in the ocean. The scale of this heterogeneity spans many orders of magnitude in its spatial extent, ranging from planetary down to microscale. It has been argued that patchiness plays a fundamental role in the functioning of marine ecosystems, insofar as the mean conditions may not reflect the environment to which organisms are adapted. Understanding the nature of this patchiness is thus one of the major challenges of oceanographic ecology. The patchiness problem is fundamentally one of physical-biological-chemical interactions. This interconnection arises from three basic sources: (1) ocean currents continually redistribute dissolved and suspended constituents by advection; (2) space-time fluctuations in the flows themselves impact biological and chemical processes, and (3) organisms are capable of directed motion through the water. This tripartite linkage poses a difficult challenge to understanding oceanic ecosystems: differentiation between the three sources of variability requires accurate assessment of property distributions in space and time, in addition to detailed knowledge of organismal repertoires and the processes by which ambient conditions control the rates of biological and chemical reactions. Various methods of observing the ocean tend to lie parallel to the axes of the space/time domain in which these physical-biological-chemical interactions take place. Given that a purely observational approach to the patchiness problem is not tractable with finite resources, the coupling of models with observations offers an alternative which provides a context for synthesis of sparse data with articulations of fundamental principles assumed to govern functionality of the system. In a sense, models can be used to fill the gaps in the space/time domain, yielding a framework for exploring the controls on spatially and temporally intermittent processes. The following discussion highlights only a few of the multitude of models which have yielded insight into the dynamics of plankton patchiness. In addition, this particular collection of examples is intended to furnish some exposure to the diversity of modeling approaches which can be brought to bear on the problem. These approaches range from abstract theoretical models intended to elucidate specific processes, to complex numerical formulations which can be used to actually simulate observed distributions in detail.

McGillicuddy, D. J.↗

NASA NTRS2002

Evaluation of Biofilms and the Effects of Biocides Thereon

Biofilm formation is monitored by real-time continuous measurement. Images are formed of sessile cells on a surface and planktonic cells adjacent the surface. The attachment of cells to the surface is measured and quantitated, and sessile and planktonic cells are distinguished using image processing techniques. Single cells as well as colonies are monitored on or adjacent a variety of substrates. Flowing streams may be monitored. The effects of biocides on biofilms commonly isolated from recyclable water systems are measured.

Pierson, Duane L.↗

NASA NTRS2003

Variability in Ocean Color Associated with Phytoplankton and Terrigenous Matter: Time Series Measurements and Algorithm Development at the FRONT Site on the New England Continental Shelf

Fronts in the coastal ocean describe areas of strong horizontal gradients in both physical and biological properties associated with tidal mixing and freshwater estuarine output (e.g. Simpson, 1981 and O Donnell, 1993). Related gradients in optically important constituents mean that fronts can be observed from space as changes in ocean color as well as sea surface temperature (e.g., Dupouy et al., 1986). This research program is designed to determine which processes and optically important constituents must be considered to explain ocean color variations associated with coastal fronts on the New England continental shelf, in particular the National Ocean Partnership Program (NOPP) Front Resolving Observational Network with Telemetry (FRONT) site. This site is located at the mouth of Long Island sound and was selected after the analysis of 12 years of AVHRR data showed the region to be an area of strong frontal activity (Ullman and Cornillon, 1999). FRONT consists of a network of modem nodes that link bottom mounted Acoustic Doppler Current Profilers (ADCPs) and profiling arrays. At the center of the network is the Autonomous Vertically Profiling Plankton Observatory (AVPPO) (Thwaites et al. 1998). The AVPPO consists of buoyant sampling vehicle and a trawl-resistant bottom-mounted enclosure, which holds a winch, the vehicle (when not sampling), batteries, and controller. Three sampling systems are present on the vehicle, a video plankton recorder, a CTD with accessory sensors, and a suite of bio-optical sensors including Satlantic OCI-200 and OCR-200 spectral radiometers and a WetLabs ac-9 dual path absorption and attenuation meter. At preprogrammed times the vehicle is released, floats to the surface, and is then winched back into the enclosure with power and data connection maintained through the winch cable. Communication to shore is possible through a bottom cable and nearby surface telemetry buoy, equipped with a mobile modem, giving the capability for near-real time data transmission and interactive sampling control.

Morrison, John R.↗

NASA NTRS1990

Activity and adaptation of nitrilotriacetate (NTA)-degrading bacteria: field and laboratory studies

Adaptation of bacterial activity for the degradation of nitrilotriacetate (NTA) was studied using natural sediment samples and an NTA-degrading bacterium (strain ATCC 29600). Sediment samples from a river with persistent levels of NTA had much higher NTA-degradative activity than comparable samples from a less contaminated control site. When sediment from the control site was exposed to high levels of NTA a 5 day lag preceded an abrupt increase in NTA degradation while strain 29600 colonized on sand and grown in the absence of NTA became induced within eight hours. The induction of strain 29600 was compared between bacteria in suspension and cells attached to sand. The sand-associated bacteria became induced 4 to 5 h before the planktonic suspension and displayed over threefold greater specific activity. Suspensions of strain 29600 became adapted within 8 h when placed in membrane diffusion chambers that were immersed within a municipal wastewater reactor containing NTA. These findings support the concept that induction is a part of the process of bacterial adaptation to degrade NTA and sand-associated bacteria can adapt more quickly to and have a greater degradative activity for NTA than planktonic cells.

NASA Discipline Number 04-10↗

NASA NTRS2000

Climate and infectious disease: use of remote sensing for detection of Vibrio cholerae by indirect measurement

It has long been known that cholera outbreaks can be initiated when Vibrio cholerae, the bacterium that causes cholera, is present in drinking water in sufficient numbers to constitute an infective dose, if ingested by humans. Outbreaks associated with drinking or bathing in unpurified river or brackish water may directly or indirectly depend on such conditions as water temperature, nutrient concentration, and plankton production that may be favorable for growth and reproduction of the bacterium. Although these environmental parameters have routinely been measured by using water samples collected aboard research ships, the available data sets are sparse and infrequent. Furthermore, shipboard data acquisition is both expensive and time-consuming. Interpolation to regional scales can also be problematic. Although the bacterium, V. cholerae, cannot be sensed directly, remotely sensed data can be used to infer its presence. In the study reported here, satellite data were used to monitor the timing and spread of cholera. Public domain remote sensing data for the Bay of Bengal were compared directly with cholera case data collected in Bangladesh from 1992-1995. The remote sensing data included sea surface temperature and sea surface height. It was discovered that sea surface temperature shows an annual cycle similar to the cholera case data. Sea surface height may be an indicator of incursion of plankton-laden water inland, e.g., tidal rivers, because it was also found to be correlated with cholera outbreaks. The extensive studies accomplished during the past 25 years, confirming the hypothesis that V. cholerae is autochthonous to the aquatic environment and is a commensal of zooplankton, i.e., copepods, when combined with the findings of the satellite data analyses, provide strong evidence that cholera epidemics are climate-linked.

Cholera/epidemiology↗

NASA NTRS2006

Microbial Characterization of Internal Active Thermal Control System (IATCS) Hardware Surfaces after Five Years of Operation in the International Space Station

A flex hose assembly containing aqueous coolant from the International Space Station (ISS) Internal Active Thermal Control System (IATCS) consisting of a 2 foot section of Teflon hose and quick disconnects (QDs) and a Special Performance Checkout Unit (SPCU) heat exchanger containing separate channels of IATCS coolant and iodinated water used to cool spacesuits and Extravehicular Mobility Units (EMUS) were returned for destructive analyses on Shuttle return to flight mission STS-114. The original aqueous IATCS coolant used in Node 1, the Laboratory Module, and the Airlock consisted of water, borate (pH buffer), phosphate (corrosion control), and silver sulfate (microbiological control) at a pH of 9.5 +/- 0.5. Chemical changes occurred after on-orbit implementation including a decrease to pH 8.4 due to the diffusion of carbon dioxide through the Teflon hoses, an increase in nickel ions due to general corrosion of heat exchanger braze coatings, a decrease in phosphate concentration due to precipitation of nickel phosphate, and the rapid disappearance of silver ions due to deposition on hardware surfaces. Also associated with the coolant chemistry changes was an increase in planktonic microorganisms from less than 100 colony forming units (CFU) per 100 ml to approximately 1 million CFU per 100 ml. Attachment and growth of microorganisms to the system surfaces (biofilm) was suspected due to the levels of planktonic microorganisms in the coolant. Biofilms can reduce coolant flow, reduce heat transfer, amplify degradation of system materials initiated by chemical corrosion, and enhance mineral scale formation.

Roman, Monsi C.↗

NASA NTRS2017

Spaceborne Lidar in the Study of Marine Systems

Satellite passive ocean color instruments have provided an unbroken ~20-year record of global ocean plankton properties, but this measurement approach has inherent limitations in terms of spatial-temporal sampling and ability to resolve vertical structure within the water column. These limitations can be addressed by coupling ocean color data with measurements from a spaceborne lidar. Airborne lidars have been used for decades to study ocean subsurface properties, but recent breakthroughs have now demonstrated that plankton properties can be measured with a satellite lidar. The satellite lidar era in oceanography has arrived. Here we present a review of the lidar technique, its applications in marine systems, a prospective on what can be accomplished in the near future with an ocean- and atmosphere-optimized satellite lidar, and a vision for a multi-platform ‘virtual constellation’ of observational assets enabling a 3-dimensional reconstruction of global ocean ecosystems.

Chris A Hostetler↗