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Recent research has shown that microplastics are widespread in the atmosphere. However, we know little about their ability to nucleate ice and their impact on ice formation in clouds. Ice nucleation by microplastics could also limit their long-range transport and global distribution. The present study explores the heterogeneous ice-nucleating ability of seven microplastic samples in immersion freezing mode. Two polypropylene samples and one polyethylene terephthalate sample froze heterogeneously with median freezing temperatures of -20.9, -23.2, and -21.9 °C, respectively. The number of ice nucleation sites per surface area, n s ( T ), ranged from 10 -1 to 10 4 in a temperature interval of -15 to -25 °C, which is comparable to that of volcanic ash and fungal spores. After exposure to ozone or a combination of UV light and ozone, simulating atmospheric aging, the ice nucleation activity decreased in some cases and remained unchanged in others. Our freezing data suggest that microplastics may promote ice formation in cloud droplets. In addition, based on a comparison of our freezing results and previous simulations using a global transport model, ice nucleation by microplastics will impact their long-range transport to faraway locations and global distribution.-2 in a temperature interval of -15 to -25 °C, which is comparable to that of volcanic ash and fungal spores. After exposure to ozone or a combination of UV light and ozone, simulating atmospheric aging, the ice nucleation activity decreased in some cases and remained unchanged in others. Our freezing data suggest that microplastics may promote ice formation in cloud droplets. In addition, based on a comparison of our freezing results and previous simulations using a global transport model, ice nucleation by microplastics will impact their long-range transport to faraway locations and global distribution.

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., Competing Interests: The authors declare that they have no competing interests., (© 2023 Hanlon et al.)

Within the last years pollen grains have gained increasing attention due to their cloud-forming potential. Especially the discovery that ice nucleating macromolecules (INMs) or subpollen particles (SPPs) obtained from pollen grains are able to initiate freezing has stirred up interest in pollen. INMs and SPPs are much smaller and potentially more numerous than pollen grains and could significantly affect cloud formation in the atmosphere. However, INMs and SPPs are not clearly distinguished. This has motivated the present study, which focuses on birch pollen and investigates the relationship between pollen grains, INMs, and SPPs. According to the usage of the term SPP in the medical fields, we define SPPs as the starch granules contained in pollen grains. We show that these insoluble SPPs are only obtained when fresh pollen grains are used to generate aqueous extracts from pollen. Due to the limited seasonal availability of fresh pollen grains, almost all studies have been conducted with commercial pollen grains. To enable the investigation of the SPPs we develop an alternative extraction method to generate large quantities of SPPs from commercial pollen grains. We show that INMs are not bonded to SPPs (i.e. can be washed off with water). Further, we find that purified SPPs are not ice nucleation active: after several times of washing SPPs with ultrapure water the ice nucleation activity completely disappears. To our knowledge, this is the first study to investigate the ice nucleation activity of isolated SPPs. To study the chemical nature of the INMs, we use fluorescence spectroscopy. Fluorescence excitation–emission maps indicate a strong signal in the protein range (maximum around λex=280 nm and λem=330 nm) with all ice nucleation active samples. In contrast, with purified SPPs the protein signal is lost. We also quantify the protein concentration with the Bradford assay. The protein concentration ranges from 77.4 µ g mL -1 (highly concentrated INMs) to below 2.5 µ g mL -1 (purified SPPs). Moreover, we investigate the connection between proteins and ice nucleation activity by treating the ice nucleation active samples with subtilisin A and urea to unfold and digest the proteins. After this treatment the ice nucleation activity clearly diminished. The results indicate a linkage between ice nucleation activity and protein concentration. The missing piece of the puzzle could be a glycoprotein which exhibits carboxylate functionalities, can bind water in tertiary structures, and displays degeneration and unfolding of its secondary structure due to heat treatment or reaction with enzymes. Even though purified SPPs are not ice nucleation active they could act as carriers of INMs and distribute those in the atmosphere. [ABSTRACT FROM AUTHOR]

Within the last years pollen grains have gained increasing attention due to their cloud forming potential. Especially the discovery that ice nucleating macromolecules (INM) or subpollen particles (SPP) obtained from pollen grains are able to initiate freezing has stirred up interest in pollen. INM or SPP are much smaller and potentially more numerous than pollen grains and could significantly affect cloud formation in the atmosphere. However, INM and SPP are not clearly distinguished and explanations on how these materials could distribute in the atmosphere are missing. In this study we focus on birch pollen and investigate the relationship between pollen grains, INM and SPP. According to the usage of the term SPP in the medical fields we define SPP as the starch granules contained in pollen grains. We develop an extraction method to generate large quantities of SPP and show that INM are loosley attached to SPP. Further, we find that purified SPP are not ice nucleation active: after several times of washing SPP with ultrapure water the ice nucleation activity completely disappears. To our knowledge this is the first study to investigate the ice nucleation activity of isolated SPP. To study the chemical nature of the INM we use fluorescence spectroscopy. Fluorescence excitation-emission maps indicate a strong signal in the protein range (maximum around λex = 280 nm and λem = 330 nm) that correlates with the ice nucleation activity. In contrast, with purified SPP this signal is lost. We also quantify the protein concentration with the Bradford assay. The protein concentration ranges from 77.4 µg mL-1 (Highly concentrated INM) to below 2.5 µg mL-1 (purified SPP). The results indicate a linkage between ice nucleation activity and protein concentration. Even though purified SPP are not ice nucleation active they could act as carriers of INM and distribute those in the atmosphere. [ABSTRACT FROM AUTHOR]