Browsing by Author "Grothe, Hinrich"

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    Birch leaves and branches as a source of ice-nucleating macromolecules
    Felgitsch, Laura; Baloh, Philipp; Burkart, Julia; Mayr, Maximilian; Momken, Mohammad E.; Seifried, Teresa M.; Winkler, Philipp L.; Schmale, David G. III; Grothe, Hinrich (European Geosciences Union, 2018-11-08)
    Birch pollen are known to release ice-nucleating macromolecules (INM), but little is known about the production and release of INM from other parts of the tree. We examined the ice nucleation activity of samples from 10 different birch trees (Betula spp.). Samples were taken from nine birch trees in Tyrol, Austria, and from one tree in a small urban park in Vienna, Austria. Filtered aqueous extracts of 30 samples of leaves, primary wood (new branch wood, green in colour, photosynthetically active), and secondary wood (older branch wood, brown in colour, with no photosynthetic activity) were analysed in terms of ice nucleation activity using VODCA (Vienna Optical Droplet Crystallization Analyser), a cryo microscope for emulsion samples. All samples contained ice-nucleating particles in the submicron size range. Concentrations of ice nuclei ranged from 6:7 x 10⁴ to 6:1 x 10⁹ mg⁻¹ sample. Mean freezing temperatures varied between -15:6 and -31:3 °C; the range of temperatures where washes of birch pollen and dilutions thereof typically freeze. The freezing behaviour of three concentrations of birch pollen washing water (initial wash, 1 : 100, and 1 : 10 000) were significantly associated with more than a quarter of our samples, including some of the samples with highest and lowest activity. This indicates a relationship between the INM of wood, leaves, and pollen. Extracts derived from secondary wood showed the highest concentrations of INM and the highest freezing temperatures. Extracts from the leaves exhibited the highest variation in INM and freezing temperatures. Infrared spectra of the extracts and tested birch samples show qualitative similarity, suggesting the chemical components may be broadly similar.
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    Coordinated Sampling of Microorganisms Over Freshwater and Saltwater Environments Using an Unmanned Surface Vehicle (USV) and a Small Unmanned Aircraft System (sUAS)
    Powers, Craig W.; Hanlon, Regina; Grothe, Hinrich; Prussin, Aaron J. II; Marr, Linsey C.; Schmale, David G. III (Frontiers, 2018-08-15)
    Biological aerosols (bioaerosols) are ubiquitous in terrestrial and aquatic environments and may influence cloud formation and precipitation processes. Little is known about the aerosolization and transport of bioaerosols from aquatic environments. We designed and deployed a bioaerosol-sampling system onboard an unmanned surface vehicle (USV; a remotely operated boat) to collect microbes and monitor particle sizes in the atmosphere above a salt pond in Falmouth, MA, United States and a freshwater lake in Dublin, VA, United States. The bioaerosol-sampling system included a series of 3D-printed impingers, two different optical particle counters, and a weather station. A small unmanned aircraft system (sUAS; a remotely operated airplane) was used in a coordinated effort with the USV to collect microorganisms on agar media 50 m above the surface of the water. Samples from the USV and sUAS were cultured on selective media to estimate concentrations of culturable microorganisms (bacteria and fungi). Concentrations of microbes from the sUAS ranged from 6 to 9 CFU/m3 over saltwater, and 12 to 16 CFU/m3 over freshwater (over 10-min sampling intervals) at 50 m above ground level (AGL). Concentrations from the USV ranged from 0 (LOD) to 42,411 CFU/m3 over saltwater, and 0 (LOD) to 56,809 CFU/m3 over freshwater (over 30-min sampling intervals) in air near the water surface. Particle concentrations recorded onboard the USV ranged from 0 (LOD) to 288 μg/m3 for PM1, 1 to 290 μg/m3 for PM2.5, and 1 to 290 μg/m3 for PM10. A general trend of increasing concentration with an increase in particle size was recorded by each sensor. Through laboratory testing, the collection efficiency of the 3D-printed impingers was determined to be 75% for 1 μm beads and 99% for 3 μm beads. Additional laboratory tests were conducted to determine the accuracy of the miniaturized optical particle counters used onboard the USV. Future work aims to understand the distribution of bioaerosols above aquatic environments and their potential association with cloud formation and precipitation processes.
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    Diversity and ice nucleation activity of Pseudomonas syringae in drone water samples from eight lakes in Austria
    Hanlon, Regina; Jimenez-Sanchez, Celia; Benson, James; Aho, Ken; Morris, Cindy; Seifried, Teresa; Bolah, Philipp; Grothe, Hinrich; Schmale, David G. III (PeerJ, 2023-11-28)
    Bacteria from the Pseudomonas syringae complex (comprised of at least 15 recognized species and more than 60 different pathovars of P. syringae sensu stricto) have been cultured from clouds, rain, snow, streams, rivers, and lakes. Some strains of P. syringae express an ice nucleation protein (hereafter referred to as ice+) that catalyzes the heterogeneous freezing of water. Though P. syringae has been sampled intensively from freshwater sources in the U.S. and France, little is known about the genetic diversity and ice nucleation activity of P. syringae in other parts of the world. We investigated the haplotype diversity and ice nucleation activity at −8 ◦C (ice+) of strains of P. syringae from water samples collected with drones in eight freshwater lakes in Austria. A phylogenetic analysis of citrate synthase (cts) sequences from 271 strains of bacteria isolated from a semi-selective medium for Pseudomonas revealed that 69% (188/271) belonged to the P. syringae complex and represented 32 haplotypes in phylogroups 1, 2, 7, 9, 10, 13, 14 and 15. Strains within the P. syringae complex were identified in all eight lakes, and seven lakes contained ice+ strains. Partial 16S rDNA sequences were analyzed from a total of 492 pure cultures of bacteria isolated from non-selective medium. Nearly half (43.5%; 214/492) were associated with the genus Pseudomonas. Five of the lakes (ALT, GRU, GOS, GOL, and WOR) were all distinguished by high levels of Pseudomanas (p ≤ 0.001). HIN, the highest elevation lake, had the highest percentage of ice+ strains. Our work highlights the potential for uncovering new haplotypes of P. syringae in aquatic habitats, and the use of robotic technologies to sample and characterize microbial life in remote settings.
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    A Drone-Based Bioaerosol Sampling System to Monitor Ice Nucleation Particles in the Lower Atmosphere
    Bieber, Paul; Seifried, Teresa M.; Burkart, Julia; Gratzl, Jürgen; Kasper-Giebl, Anne; Schmale, David G. III; Grothe, Hinrich (MDPI, 2020-02-07)
    Terrestrial ecosystems can influence atmospheric processes by contributing a huge variety of biological aerosols (bioaerosols) to the environment. Several types of biological particles, such as pollen grains, fungal spores, and bacteria cells, trigger freezing processes in super-cooled cloud droplets, and as such can contribute to the hydrological cycle. Even though biogenic particles are known as the most active form of ice nucleation particles (INPs), the transport to high tropospheric altitudes, as well as the occurrence in clouds, remains understudied. Thus, transport processes from the land surface into the atmosphere need to be investigated to estimate weather phenomena and climate trends. To help fill this knowledge gap, we developed a drone-based aerosol particles sampling impinger/impactor (DAPSI) system for field studies to investigate sources and near surface transport of biological INPs. DAPSI was designed to attach to commercial rotary-wing drones to collect biological particles within about 100 m of the Earth’s surface. DAPSI provides information on particulate matter concentrations (PM10 & PM2.5), temperature, relative humidity, and air pressure at about 0.5 Hz, by controlling electrical sensors with an onboard computer (Raspberry Pi 3). Two remote-operated sampling systems (impinging and impacting) were integrated into DAPSI. Laboratory tests of the impinging system showed a 96% sampling efficiency for standardized aerosol particles (2 µm polystyrene latex spheres) and 84% for an aerosol containing biological INPs (Betula pendula). A series of sampling missions (12 flights) were performed using two Phantom 4 quadcopters with DAPSI onboard at a remote sampling site near Gosau, Austria. Fluorescence microscopy of impactor foils showed a significant number of auto-fluorescent particles < 0.5 µm at an excitation of 465–495 nm and an emission of 515–555 nm. A slight increase in ice nucleation activity (onset temperature between −27 °C and −31 °C) of sampled aerosol was measured by applying freezing experiments with a microscopic cooling technique. There are a number of unique opportunities for DAPSI to be used to study the transport of bioaerosols, particularly for investigations of biological INP emissions from natural sources such as birch or pine forests.
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    Ice Nucleation Activity of Alpine Bioaerosol Emitted in Vicinity of a Birch Forest
    Seifried, Teresa M.; Bieber, Paul; Kunert, Anna T.; Schmale, David G. III; Whitmore, Karin; Fröhlich-Nowoisky, Janine; Grothe, Hinrich (MDPI, 2021-06-17)
    In alpine environments, many plants, bacteria, and fungi contain ice nuclei (IN) that control freezing events, providing survival benefits. Once airborne, IN could trigger ice nucleation in cloud droplets, influencing the radiation budget and the hydrological cycle. To estimate the atmospheric relevance of alpine IN, investigations near emission sources are inevitable. In this study, we collected 14 aerosol samples over three days in August 2019 at a single site in the Austrian Alps, close to a forest of silver birches, which are known to release IN from their surface. Samples were taken during and after rainfall, as possible trigger of aerosol emission by an impactor and impinger at the ground level. In addition, we collected aerosol samples above the canopy using a rotary wing drone. Samples were analyzed for ice nucleation activity, and bioaerosols were characterized based on morphology and auto-fluorescence using microscopic techniques. We found high concentrations of IN below the canopy, with a freezing behavior similar to birch extracts. Sampled particles showed auto-fluorescent characteristics and the morphology strongly suggested the presence of cellular material. Moreover, some particles appeared to be coated with an organic film. To our knowledge, this is the first investigation of aerosol emission sources in alpine vegetation with a focus on birches.
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    Microorganisms Collected from the Surface of Freshwater Lakes Using a Drone Water Sampling System (DOWSE)
    Benson, Jamie; Hanlon, Regina; Seifried, Teresa M.; Baloh, Philipp; Powers, Craig W.; Grothe, Hinrich; Schmale, David G. III (MDPI, 2019-01-16)
    New tools and technology are needed to study microorganisms in freshwater environments. Little is known about spatial distribution and ice nucleation activity (INA) of microorganisms in freshwater lakes. We developed a system to collect water samples from the surface of lakes using a 3D-printed sampling device tethered to a drone (DOWSE, DrOne Water Sampling SystEm). The DOWSE was used to collect surface water samples at different distances from the shore (1, 25, and 50 m) at eight different freshwater lakes in Austria in June 2018. Water samples were filtered, and microorganisms were cultured on two different media types, TSA (a general growth medium) and KBC (a medium semi-selective for bacteria in the genus Pseudomonas). Mean concentrations (colony forming units per mL, or CFU/mL) of bacteria cultured on TSA ranged from 19,800 (Wörthersee) to 210,500 (Gosaulacke) CFU/mL, and mean concentrations of bacteria cultured on KBC ranged from 2590 (Ossiachersee) to 11,000 (Vorderer Gosausee) CFU/mL. There was no significant difference in sampling distance from the shore for concentrations of microbes cultured on TSA (p = 0.28). A wireless bathymetry sensor was tethered to the drone to map temperature and depth across the sampling domain of each of the lakes. At the 50 m distance from the shore, temperature ranged from 17 (Hinterer Gosausee, and Gosaulacke) to 26 °C (Wörthersee), and depth ranged from 2.8 (Gosaulacke) to 11.1 m (Grundlsee). Contour maps of concentrations of culturable bacteria across the drone sampling domain revealed areas of high concentrations (hot spots) in some of the lakes. The percentage of ice-nucleation active (ice+) bacteria cultured on KBC ranged from 0% (0/64) (Wörthersee) to 58% (42/72) (Vorderer Gosausee), with a mean of 28% (153/544) for the entire sample set. Future work aims to elucidate the structure and function of entire microbial assemblages within and among the Austrian lakes.
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    Perspectives on the Future of Ice Nucleation Research: Research Needs and Unanswered Questions Identified from Two International Workshops
    Coluzza, Ivan; Creamean, Jessie; Rossi, Michel J.; Wex, Heike; Alpert, Peter Aaron; Bianco, Valentino; Boose, Yvonne; Dellago, Christoph; Felgitsch, Laura; Fröhlich-Nowoisky, Janine; Herrmann, Hartmut; Jungblut, Swetlana; Kanji, Zamin A.; Menzl, Georg; Moffett, Bruce; Moritz, Clemens; Mutzel, Anke; Pöschl, Ulrich; Schauperl, Michael; Scheel, Jan; Stopelli, Emiliano; Stratmann, Frank; Grothe, Hinrich; Schmale, David G. III (MDPI, 2017-07-29)
    There has been increasing interest in ice nucleation research in the last decade. To identify important gaps in our knowledge of ice nucleation processes and their impacts, two international workshops on ice nucleation were held in Vienna, Austria in 2015 and 2016. Experts from these workshops identified the following research needs: (1) uncovering the molecular identity of active sites for ice nucleation; (2) the importance of modeling for the understanding of heterogeneous ice nucleation; (3) identifying and quantifying contributions of biological ice nuclei from natural and managed environments; (4) examining the role of aging in ice nuclei; (5) conducting targeted sampling campaigns in clouds; and (6) designing lab and field experiments to increase our understanding of the role of ice-nucleating particles in the atmosphere. Interdisciplinary teams of scientists should work together to establish and maintain a common, unified language for ice nucleation research. A number of commercial applications benefit from ice nucleation research, including the production of artificial snow, the freezing and preservation of water-containing food products, and the potential modulation of weather. Additional work is needed to increase our understanding of ice nucleation processes and potential impacts on precipitation, water availability, climate change, crop health, and feedback cycles.
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    Seasonal ice nucleation activity of water samples from alpine rivers and lakes in Obergurgl, Austria
    Baloh, Philipp; Hanlon, Regina; Anderson, Christopher; Dolan, Eoin; Pacholik, Gernot; Stinglmayr, David; Burkart, Julia; Felgitsch, Laura; Schmale, David G. III; Grothe, Hinrich (Elsevier, 2021-07-30)
    Heterogeneous ice nucleation plays an important role in many environmental processes such as ice cloud formation, freezing of water bodies or biological freeze protection in the cryosphere. New information is needed about the seasonal availability, nature, and activity of ice nucleating particles (INPs) in alpine environments. These INPs trigger the phase transition from liquid water to solid ice at elevated subzero temperatures. We collected water samples from a series of alpine rivers and lakes (two valleys and their rivers, an artificial pond, and a natural lake system) in Obergurgl, Austria in June 2016, July 2016, November 2016, and May 2017. Each alpine river and lake was sampled multiple times across different seasons, depending on site access during different times of the year. Water samples were filtered through a 0.22 μm membrane filter to separate microbial INPs from the water, and both fractions were analyzed for ice nucleation activity (INA) by an emulsion freezing method. Microorganisms were cultured from the filters, and the cultures then analyzed for INA. Portions of the filtered samples were concentrated by lyophilization to observe potential enhancement of INA. Two sediment samples were taken as reference points for inorganic INPs. Sub-micron INPs were observed in all of the alpine water sources studied, and a seasonal shift to a higher fraction of microbial ice nucleators cultured on selective media was observed during the winter collections. Particles larger than 0.22 μm showed INA, and microbes were cultured from this fraction. Results from 60 samples gave evidence of a seasonal change in INA, presence of submicrometer INPs, and show the abundance of culturable microorganisms, with late spring and early summer showing the most active biological INPs. With additional future research on this topic ski resorts could make use of such knowledge of geographical and seasonal trends of microbial INPs in freshwater habitats in order to improve the production of artificial snow.
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    Surfaces of silver birch (Betula pendula) are sources of biological ice nuclei: in vivo and in situ investigations
    Seifried, Teresa M.; Bieber, Paul; Felgitsch, Laura; Vlasich, Julian; Reyzek, Florian; Schmale, David G. III; Grothe, Hinrich (2020-11-20)
    Silver birch (Betula pendula) is known to contain ice-nucleating macromolecules (INMs) to survive in harsh environments. However, little is known about the release and transport of INMs from birch trees into the atmosphere. In this study, we conducted in situ and in vivo investigations on INMs from nine birches growing in an alpine valley (Otztal, Austria). A detailed analysis of drill cores showed that INM concentration increases towards outer layers, reaching its maximum near the surface. Aqueous extracts from the surfaces of leaves, bark, primary wood and secondary wood contained INMs (34/36) with concentrations ranging from 9.9 x 10(5) to 1.8 x 10(9) INMs cm(-2). In a field study, we analysed the effect of precipitation on the release of these INMs attached to the surface of the trees. These experiments showed that INMs are splashed and aerosolized into the environment during rainfall events, at concentrations and freezing temperatures similar to in vivo samples. Our work sheds new light on the release and transport of INMs from birch surfaces into the troposphere. Birches growing in boreal and alpine forests should be considered an important terrestrial source of INMs.
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    Wind-driven spume droplet production and the transport of Pseudomonas syringae from aquatic environments
    Pietsch, Renee B.; Grothe, Hinrich; Hanlon, Regina; Powers, Craig W.; Jung, Sunghwan; Ross, Shane D.; Schmale, David G. III (PeerJ, 2018-09-26)
    Natural aquatic environments such as oceans, lakes, and rivers are home to a tremendous diversity of microorganisms. Some may cross the air-water interface within droplets and become airborne, with the potential to impact the Earth’s radiation budget, precipitation processes, and spread of disease. Larger droplets are likely to return to the water or adjacent land, but smaller droplets may be suspended in the atmosphere for transport over long distances. Here, we report on a series of controlled laboratory experiments to quantify wind-driven droplet production from a freshwater source for low wind speeds. The rate of droplet production increased quadratically with wind speed above a critical value (10-m equivalent 5.7 m/s) where droplet production initiated. Droplet diameter and ejection speeds were fit by a gamma distribution. The droplet mass flux and momentum flux increased with wind speed. Two mechanisms of droplet production, bubble bursting and fragmentation, yielded different distributions for diameter, speed, and angle. At a wind speed of about 3.5 m/s, aqueous suspensions of the ice-nucleating bacterium Pseudomonas syringae were collected at rates of 283 cells m−2 s−1 at 5 cm above the water surface, and at 14 cells m−2 s−1 at 10 cm above the water surface. At a wind speed of about 4.0 m/s, aqueous suspensions of P. syringae were collected at rates of 509 cells m−2 s−1 at 5 cm above the water surface, and at 81 cells m−2 s−1 at 10 cm above the water surface. The potential for microbial flux into the atmosphere from aquatic environments was calculated using known concentrations of bacteria in natural freshwater systems. Up to 3.1 × 104 cells m−2 s−1 of water surface were estimated to leave the water in potentially suspended droplets (diameters <100 µm). Understanding the sources and mechanisms for bacteria to aerosolize from freshwater aquatic sources may aid in designing management strategies for pathogenic bacteria, and could shed light on how bacteria are involved in mesoscale atmospheric processes.