Data Files Accompanying: Cultivable, halotolerant ice nucleating bacteria and fungi in coastal precipitation
Authors:  Charlotte M. Beall, Jennifer M. Michaud, Meredith A. Fish, Julie Dinasquet, Gavin C. Cornwell, M. Dale Stokes, Michael D. Burkart, Thomas C. Hill, Paul J. DeMott, Kimberly A. Prather
Journal: Atmospheric Chemistry and Physics

Contact: Kimberly A. Prather, kprather@ucsd.edu, Scripps Institution of Oceanography, Department of Chemistry & Biochemistry, University of California San Diego

Cite as: Beall, Charlotte M.; Michaud, Jennifer M.; Fish, Meredith A.; Stokes, M. Dale; Burkart, Michael D.; Prather, Kimberly A. (2020). Data from: Cultivable, halotolerant ice nucleating bacteria and fungi in coastal precipitation. In Center for Aerosol Impacts on Chemistry of the Environment (CAICE). UC San Diego Library Digital Collections. https://doi.org/10.6075/J0GQ6W2Z  

Contents of this directory include the following:

Subdirectory “IsoZoBell” – CSVs featuring INP data for all isolates suspended in ZoBell growth media.  Filenames correspond to the isolate ID.  
Subdirectory “ZoBell” – CSVs featuring INP data for each ZoBell sample. Filenames correspond to the ZoBell sample ID.
Subdirectory "IsoZoBell_Biomass" - CSVs featuring estimates of INP per g biomass for isolates that were determined to have significant freezing activity above the respective ZoBell sample. 
Subdirectory “IsoFASW” – CSVs featuring INP data for the subset of isolates that were washed and resuspended in Filtered Autoclaved SeaWater (FASW).  Filenames correspond to the isolate ID.
Subdirectory “FASW” – CSVs featuring INP data for each FASW sample.  Filenames correspond to the FASW sample ID.
CloudConditions.csv – contains estimates of the precipitating cloud altitude for each sampling period, derived from the High-Resolution Rapid Refresh model (HRRR).  Minimum, maximum, and average altitudes were used as release points for FLEXPART back-trajectory simulations.
inIsolates_dataDictionary.csv – contains descriptions and formatting of each feature in this dataset
PrecipSamplingTimes.csv – contains dates and times that correspond to each precipitation sampling period
AerosolSamplingTimes.csv – contains dates and times that correspond to each aerosol sampling period
TableS3Taxa.csv – contains summary of ice nucleation test results, 16S BLAST sequences, % identity with BLAST assignment and isolate taxonomy

Details on data collection:
Within one to two hours of collection, precipitation and aerosol samples were inoculated in 5 mL ZoBell growth media (5 g peptone, 1 g yeast extract per 1 L) in filtered (0.22 µm) autoclaved seawater and grown under ambient conditions (21 - 24 ˚C).Isolation was performed by successive plating on ZoBell agar. Liquid cultures were inoculated from single colonies and grown to late exponential phase. DNA was extracted from liquid cultures of isolates after an overnight lysis with proteinaseK and lysozyme (MilliporeSigma, Burlington, Massachusetts, USA) using a QIAamp® kit (QIAGEN, Hilden, Germany). 16S V4 ribosomal DNA fragments were amplified using the primers 515F (GTGYCAGCMGCCGCGGTAA) and 926R (CCGYCAATTCMTTTRAGT). PCR products were purified using GenEluteTM PCR Clean-up kit (MilliporeSigma). 16S fragment DNA sequences were resolved by sanger sequencing (Retrogen, San Diego, CA). OTUs were determined from 16S sequences using SINA and individual sequences were inspected using BLAST (https://www.ncbi.nlm.nih.gov/) for further characterization. To investigate the possible influence from ZoBell growth media on INP measurements, a subset of late exponential cultures were washed three times with filtered (0.22 µm) autoclaved seawater (FASW) by successive centrifugation and resuspension. 

INP measurements were made using the SIO-Automated Ice Spectrometer, an automated offline freezing assay technique for measurement of immersion mode INPs. 

Cloud altitudes at the time of precipitation sample collection were estimated using the High-Resolution Rapid Refresh model (HRRR). The altitudes and pressure levels of clouds were assumed to be located where RH > 95-100 % in the model. Three altitudes of the estimated cloud top, middle and bottom were used as release points of FLEXPART 10-day LaGrangian backward trajectories. Back-trajectories were used to identify potential sources of INPs in the precipitation samples, and to indicate potential sources of land-based contamination in aerosol and precipitation samples due to local wind patterns or land-sea breezes.