Prevalence and characterization of Ice Nucleation Active (INA) bacteria from rainwater in Indonesia.

BACKGROUND: Ice nucleation active (INA) bacteria are a group of microorganisms that can act as biological nucleator due to their ice nucleation protein property. Unfortunately, little is known about their prevalence and characteristics in tropical areas including Indonesia. Here, we monitor the presence of INA bacteria in rainwater and air samples collected from Jakarta, Tangerang and several areas in Western Java, Indonesia for one year. We further identify and characterize selected Class A of INA bacteria isolated from these areas. RESULTS: Most of the INA bacteria were isolated from rainwater samples collected during March-August 2010, particularly from Jakarta, Bandung, and Tangerang. A total of 1,902 bacterial isolates were recovered from these area. We found a limited number of bacterial isolates from air sampling. From ice nucleation activity assays, 101 INA isolates were found active as ice nucleator at a temperature above -10 °C. A large majority (73 isolates) of them are classified as Class C (active below -8 °C), followed by Class A (26 isolates; active at -2 to -5 °C) and Class B (two isolates; active at -5 to -8 °C). We sequenced the 16S rRNA gene of 18 Class A INA isolates and identified 15 isolates as Enterobacteriaceae, while the remaining three as Pseudomonadaceae. The vast majority of our Class A INA isolates were likely Pantoea spp. with several isolates were deduced as either Pseudomonas, Cronobacter, and Klebsiella. We found that these 18 Class A INA isolates had acquired resistance to antibiotics erythromycin and ampicillin, which are considered two critically important antibiotics. CONCLUSIONS: Our results showed that the prevalence of INA bacterial population varies across locations and seasons. Furthermore, our isolates were dominated by Class A and C INA bacteria. This study also cautions regarding the spread of antibiotic resistance among INA bacteria.

[Physical Basis of Functioning of Antifreeze Protein].

Antifreeze proteins, expressed in cold-blooded organisms, prevent ice formation in their bodies, and thus help them to survive in extremely cold winter temperatures. However, the mechanism of action of these proteins is still not clear. In any case, it is not simply a decrease in the temperature of normal ice formation. In this work, investigating the ice-binding protein (a mutant form of the antifreeze protein cfAFP from the spruce budworm Choristoneura fumiferana, which overwinters in needles), we showed that this antifreeze protein does not at all lower the freezing point of water and, paradoxically, increases the melting point of ice. On the other hand, calculations based on the theory of crystallization show that at temperatures of 0° to -30°C ice can only appear on surfaces that contact water, but not in the body of water. These facts suggest a new perspective on the role of antifreeze proteins: their task is not (as it is commonly believed) to bind with nascent ice crystals already formed in the organism and stop their growth, but to bind to those surfaces, on which ice nuclei can appear, and thus completely inhibit the ice formation in supercooled water or biological fluid.

Biological Ice-Nucleating Particles Deposited Year-Round in Subtropical Precipitation.

Airborne bacteria that nucleate ice at relatively warm temperatures (>-10°C) can interact with cloud water droplets, affecting the formation of ice in clouds and the residency time of the cells in the atmosphere. We sampled 65 precipitation events in southeastern Louisiana over 2 years to examine the effect of season, meteorological conditions, storm type, and ecoregion source on the concentration and type of ice-nucleating particles (INPs) deposited. INPs sensitive to heat treatment were inferred to be biological in origin, and the highest concentrations of biological INPs (∼16,000 INPs liter-1 active at ≥-10°C) were observed in snow and sleet samples from wintertime nimbostratus clouds with cloud top temperatures as warm as -7°C. Statistical analysis revealed three temperature classes of biological INPs (INPs active from -5 to -10°C, -11 to -12°C, and -13 to -14°C) and one temperature class of INPs that were sensitive to lysozyme (i.e., bacterial INPs, active from -5 to -10°C). Significant correlations between the INP data and abundances of taxa in the Bacteroidetes, Firmicutes, and unclassified bacterial divisions implied that certain members of these phyla may possess the ice nucleation phenotype. The interrelation between the INP classes and fluorescent dissolved organic matter, major ion concentrations (Na+, Cl-, SO42-, and NO3-), and backward air mass trajectories indicated that the highest concentrations of INPs were sourced from high-latitude North American and Asian continental environments, whereas the lowest values were observed when air was sourced from marine ecoregions. The intra- and extracontinental regions identified as sources of biological INPs in precipitation deposited in the southeastern United States suggests that these bioaerosols can disperse and affect meteorological conditions thousands of kilometers from their terrestrial points of origin.IMPORTANCE The particles most effective at inducing the freezing of water in the atmosphere are microbiological in origin; however, information on the species harboring this phenotype, their environmental distribution, and ecological sources are very limited. Analysis of precipitation collected over 2 years in Louisiana showed that INPs active at the warmest temperatures were sourced from terrestrial ecosystems and displayed behaviors that implicated specific bacterial taxa as the source of the ice nucleation activity. The abundance of biological INPs was highest in precipitation from winter storms and implied that their in-cloud concentrations were sufficient to affect the formation of ice and precipitation in nimbostratus clouds.

Forest tree pollen dispersal via the water cycle.

PREMISE OF THE STUDY: Pine pollen (Pinus spp.), along with other atmospheric particles, is dispersed by the water cycle, but this mode of dispersal requires cloud-pollen interactions that depend on taxon-specific biological properties. In the simplest form of this dispersal, pine pollen ascends vertically to altitudes of 2 to 6 km, where a fraction is captured by mixed-phase cloud formation. Captured pollen accretes into frozen droplets, which ultimately descend as rain, snow, or hail. Whether Pinus pollen can still germinate after its exposure to high-altitude freezing is pertinent to (1) how forests adapt to climate change and (2) potential gene flow between genetically modified plantation species and their conspecific relatives. • METHODS: To address this question, pollen from four Old World and two New World Pinus species were subjected to immersion freezing, a common cloud formation mode, under laboratory conditions. • KEY RESULTS: Some pollen grains immersed at -20°C for 15, 60, or 120 min in either a dehydrated or a water-saturated state were still capable of germination. After exposure, dehydrated pine pollen had higher germination (43.3%) than water-saturated pollen (7.6%). • CONCLUSIONS: Pine pollen exposed to freezing during cloud formation can still germinate, raising the question of whether rain-delivered live pollen might be linked to rain-facilitated pollination. Dispersal of live pine pollen via cloud formation and the water cycle itself deserves closer study.

On-line measurement of fluorescent aerosols near an industrial zone in the Yangtze River Delta region using a wideband integrated bioaerosol spectrometer.

In this work, we present on-line fluorescent aerosol measurements by the wideband integrated bioaerosol spectrometer (WIBS-4A) near an industrial zone in Nanjing, a megacity in the Yangtze-River-Delta (YRD) region. The fieldwork was conducted from April 1 to May 8, 2014. A TSI. 3321 aerosol-particle-sizer (APS) was simultaneously deployed to measure the total number size distribution of aerosol with diameter from 0.8-20 µm. Both WIBS-4A and APS reported similar number concentration and temporal profiles (R2 = 0.72). However, the daily average number of potential bioaerosols was only 0.5 ±â€¯0.2% of the total particles detected by the WIBS-4A and displayed a completely different diurnal profile from that of APS. In addition, WIBS-4A can only provide integrated fluorescent signals, which strongly limited the potential to specifically identify the bioaerosols. Accordingly, hierarchical-agglomerative-cluster-analysis (HACA) was utilized to identify and speciate the potential bioaerosols from the WIBS-4A dataset. By maximizing the total distances among all potential cluster centers, a 12-cluster solution was accepted as the optimum result. These clusters were further identified according to their fluorescent signatures, size, and morphology, i.e., non-bioaerosols, bacteria, and fungal spores and/or pollen fragments. Bacteria were the dominant bioaerosol species detected in this work. The diurnal profiles of bioaerosols correlated very well with relatively humidity (RH), reaching daily maxima around 3 AM~6 AM, indicating the presence of humidity controlled bioaerosol emission mechanism, i.e., bacteria may flourish under moderate ambient temperature, RH, and the absence of UV radiation. The size- and AF-distributions of bioaerosols indicated that bioaerosols normally varied substantially in size and assumed a rather irregular shape. Although the number concentration of bioaerosols was relatively small, most bioaerosols can efficiently serve as ice nuclei by providing rough and irregular surfaces, verified by the observation results. Therefore, WIBS-4A measurements can still be informative for investigations of bioaerosols in the atmosphere, especially when HACA method was incorporated into the data processing.

Anthropogenic Aerosol Indirect Effects in Cirrus Clouds.

We have implemented a parameterization for forming ice in large-scale cirrus clouds that accounts for the changes in updrafts associated with a spectrum of waves acting within each time step in the model. This allows us to account for the frequency of homogeneous and heterogeneous freezing events that occur within each time step of the model and helps to determine more realistic ice number concentrations as well as changes to ice number concentrations. The model is able to fit observations of ice number at the lowest temperatures in the tropical tropopause but is still somewhat high in tropical latitudes with temperatures between 195°K and 215°K. The climate forcings associated with different representations of heterogeneous ice nuclei (IN or INPs) are primarily negative unless large additions of IN are made, such as when we assumed that all aircraft soot acts as an IN. However, they can be close to zero if it is assumed that all background dust can act as an INP irrespective of how much sulfate is deposited on these particles. Our best estimate for the forcing of anthropogenic aircraft soot in this model is -0.2 ± 0.06 W/m2, while that from anthropogenic fossil/biofuel soot is -0.093 ± 0.033 W/m2. Natural and anthropogenic open biomass burning leads to a net forcing of -0.057 ± 0.05 W/m2.

Biophysical characterization of soluble Pseudomonas syringae ice nucleation protein InaZ fragments.

Ice nucleation protein (INP) with its functional domain consisting of multiple 48-residue repeat units effectively induces super-cooled water into ice. Circular dichroism and infrared deconvolution analyses on a soluble 240-residue fragment of Pseudomonas syringae InaZ (InaZ240) containing five 48-residue repeat units indicated that it is mostly composed of ß-sheet and random coil. Analytical ultracentrifugation suggested that InaZ240 behaves as a monomer of an elongated ellipsoid. However, InaZ240 showed only minimum ice binding compared to anti-freeze proteins. Other P. syringae InaZ proteins with more 48-residue repeat units were made, in which the largest soluble fragment obtainable was an InaZ with twelve 48-residue repeat units. Size-exclusion chromatography analyses further suggested that the overall shape of the expressed InaZ fragments is pH-dependent, which becomes compact as the numbers of 48-residue repeat unit increase.

Novel aerosol analysis approach for characterization of nanoparticulate matter in snow.

Tropospheric aerosols are involved in several key atmospheric processes: from ice nucleation, cloud formation, and precipitation to weather and climate. The impact of aerosols on these atmospheric processes depends on the chemical and physical characteristics of aerosol particles, and these characteristics are still largely uncertain. In this study, we developed a system for processing and aerosolization of melted snow in particle-free air, coupled with a real-time measurement of aerosol size distributions. The newly developed technique involves bringing snow-borne particles into an airborne state, which enables application of high-resolution aerosol analysis and sampling techniques. This novel analytical approach was compared to a variety of complementary existing analytical methods as applied for characterization of snow samples from remote sites in Alert (Canada) and Barrow (USA), as well as urban Montreal (Canada). The dry aerosol measurements indicated a higher abundance of particles of all sizes, and the 30 nm size dominated in aerosol size distributions for the Montreal samples, closely followed by Barrow, with about 30% fewer 30 nm particles, and about four times lower 30 nm particle abundance in Alert samples, where 15 nm particles were most abundant instead. The aerosolization technique, used together with nanoparticle tracking analysis and electron microscopy, allowed measurement of a wide size range of snow-borne particles in various environmental snow samples. Here, we discuss the application of the new technique to achieve better physicochemical understanding of atmospheric and snow processes. The results showed high sensitivity and reduction of particle aggregation, as well as the ability to measure a high-resolution snow-borne particle size distribution, including nanoparticulate matter in the range of 10 to 100 nm.