The bacterium Pseudomonas syringae is found in a variety of terrestrial and aquatic environments. Some strains of P. syringae express an ice nucleation protein (hereafter referred to as Ice+) allowing them to catalyze the heterogeneous freezing of water. Though P. syringae has been sampled intensively from freshwater sources in France, little is known about the genetic diversity of P. syringae in natural aquatic habitats in North America. We collected samples of freshwater from three different depths in Claytor Lake, Virginia, USA between November 2015 and June 2016. Samples were plated on non-selective medium (TSA) and on medium selective for Pseudomonas (KBC) and closely related species to estimate the total number of culturable bacteria and of Pseudomonas, respectively. A droplet freezing assay was used to screen colonies for the Ice+ phenotype. Ice+ colonies were then molecularly identified based on the cts (citrate synthase) gene and the 16S rDNA gene. Phylogenetic analysis of cts sequences showed a surprising diversity of phylogenetic subgroups of P. syringae. Frequencies of Ice+ isolates on P. syringae selective medium ranged from 0 to 15% per sample with the highest frequency being found in spring. Our work shows that freshwater lakes can be a significant reservoir of Ice+ P. syringae. Future work is needed to determine the contribution of P. syringae from freshwater lakes to the P. syringae populations present in the atmosphere and on plants and, in particular, if freshwater lakes could be an inoculum source of P. syringae-caused plant disease outbreaks.

The body has approximately 434 muscles, which makes up 40-50% of the body by weight. Muscle is hierarchical in nature and organized in progressively larger units encased in connective tissue. Like many soft tissues, muscle has nonlinear visco-elastic behavior, but muscle also has unique characteristics of excitability and contractibility. Mechanical testing of muscle has been done for crash models, pressure sore models, back pain, and other disease models. The majority of previous biomechanical studies on muscle have been associated with tensile properties in the longitudinal direction as this is muscle's primary mode of operation under normal physiological conditions. Injury conditions, particularly high rate injuries, can expose muscle to multiple stress states. Compressive stresses can lead to tissue damage, which may not be reversible. In this study, we evaluate the structure-property relationships of porcine muscle tissue under compression, in both the transverse and longitudinal orientations at 0.1 s-1, 0.01 s-1, or 0.001 s-1. Our results show an initial toe region followed by an increase in stress for muscle in both the longitudinal and transverse directions tested to 50% strain. Strain rate dependency was also observed with the higher strain rates showing significantly more stress at 50% strain. Muscle in the transverse orientation was significantly stiffer than in the longitudinal orientation indicating anisotropy. The mean area of fibers in the longitudinal orientation shows an increasing mean fiber area and a decreasing mean fiber area in the transverse orientation. Data obtained in this study can help provide insight on how muscle injuries are caused, ranging from low energy strains to high rate blast events, and can also be used in developing computational injury models.

The aerosolization of microorganisms from aquatic environments is understudied. This article describes a study in which an ice nucleation active (iceC) strain and a nonice nucleation active (ice¡) strain of the bacterium Pseudomonas syringae were aerosolized from aqueous suspensions under artificial laboratory conditions using a collison nebulizer. The aerosolization of P. syringae was not influenced by water temperatures between 5 and 30 C. In general, the culturability (viability) of P. syringae in aerosols increased with temperature between 5 and 30 C. The iceC strain was aerosolized in greater numbers than the ice-strain at all temperatures studied, suggesting a possible connection between the ice nucleation phenotype and aerosol production. Together, our results suggest that P. syringae has the potential to be aerosolized from natural aquatic environments, such as streams, rivers, ponds, and lakes; known reservoirs of P. syringae. Future work is needed to elucidate the mechanisms of aerosolization of P. syringae from natural aquatic systems.

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.

Understanding the behavior of skeletal muscle is critical to implementing computational methods to study how the body responds to compressive loading. This work presents a novel approach to studying the fully nonlinear response of skeletal muscle in compression. Porcine muscle was compressed in both the longitudinal and transverse directions under five stress relaxation steps. Each step consisted of 5% engineering strain over 1 s followed by a relaxation period until equilibrium was reached at an observed change of 1 g/min. The resulting data were analyzed to identify the peak and equilibrium stresses as well as relaxation time for all samples. Additionally, a fully nonlinear strain energy density-based Prony series constitutive model was implemented and validated with independent constant rate compressive data. A nonlinear least squares optimization approach utilizing the Levenberg-Marquardt algorithm was implemented to fit model behavior to experimental data. The results suggested the time-dependent material response plays a key role in the anisotropy of skeletal muscle as increasing strain showed differences in peak stress and relaxation time (p < 0.05), but changes in equilibrium stress disappeared (p > 0.05). The optimizing procedure produced a single set of hyper-viscoelastic parameters which characterized compressive muscle behavior under stress relaxation conditions. The utilized constitutive model was the first orthotropic, fully nonlinear hyper-viscoelastic model of skeletal muscle in compression while maintaining agreement with constitutive physical boundaries. The model provided an excellent fit to experimental data and agreed well with the independent validation in the transverse direction.