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Fungi such as Candida albicans exist in biofilm phenotypes, which present as viscoelastic materials; however, a method to measure linear viscoelastic moduli, yield stress, and yield strain is lacking. Characterization methods for fungal materials have been limited to techniques specific to particular industries. Here, we present a method to measure the shear stress, strain amplitude, and creep of C. albicans BWP17 biofilms. Our method includes features tailored to the analysis of fungi including an in vitro growth protocol attuned to the slow growth rates of C. albicans biofilms and a resultant cultured biofilm that has sufficient integrity to be transferred to the rheometer tooling without disrupting its structure. The method's performance is demonstrated by showing that results are insensitive to gap, evaporative sealant, length of experiment, and specimen radius. Multiscale imaging of the fungal biofilm showed complex entanglement networks at the hundred-micrometer scale. For a wild-type strain cultivated for 14 days, using small-amplitude oscillatory rheology, we found that the elastic (G′) and viscous (G″) moduli were nearly independent of frequency over the range 0.1–10 s −1 , with magnitudes of [Formula: see text] and [Formula: see text], respectively. The yield stress was approximately [Formula: see text]. We modeled the linear creep response of the fungal biofilm and found that C. albicans has a characteristic relaxation time of [Formula: see text] and a viscosity of [Formula: see text]. We applied this method to probe the effects of altered chitin deposition in the C. albicans cell wall. Differences between the biofilm's phenotypic cell shape and rheological properties in mutants with altered chitin synthase activity were resolved. Discovering how genotypic, phenotypic, and environmental factors impact the material properties of these microbial communities can have implications for understanding fungal biofilm growth and aid in the development of remediation strategies. 

Colloidal gels represent an important class of soft matter, in which networks formed due to strong, short-range interactions display solid-like mechanical properties, such as a finite low-frequency elastic modulus. Here we examine the effect of embedded active colloids on the linear viscoelastic moduli of fractal cluster colloidal gels. We find that the autonomous, out-of-equilibrium dynamics of active colloids incorporated into the colloidal network decreases gel elasticity, in contrast to observed stiffening effects of myosin motors in actin networks. Fractal cluster gels are formed by the well-known mechanism of aggregating polystyrene colloids through addition of divalent electrolyte. Active Janus particles with a platinum hemisphere are created from the same polystyrene colloids and homogeneously embedded in the gels at dilute concentration at the time of aggregation. Upon addition of hydrogen peroxide – a fuel that drives the diffusiophoretic motion of the embedded Janus particles – the microdynamics and mechanical rheology change in proportion to the concentration of hydrogen peroxide and the number of active colloids. We propose a theoretical explanation of this effect in which the decrease in modulus is mediated by active motion-induced softening of the inter-particle attraction. Furthermore, we characterize the failure of the fluctuation–dissipation theorem in the active gels by identifying a discrepancy between the frequency-dependent macroscopic viscoelastic moduli and the values predicted by microrheology from measurement of the gel microdynamics. These findings support efforts to engineer gels for autonomous function by tuning the microscopic dynamics of embedded active particles. Such reconfigurable gels, with multi-state mechanical properties, could find application in materials such as paints and coatings, pharmaceuticals, self-healing materials, and soft robotics. 

Abstract Evolution in similar environments often leads to convergence of behavioral and anatomical traits. A classic example of convergent trait evolution is the reduced traits that characterize many cave animals: reduction or loss of pigmentation and eyes. While these traits have evolved many times, relatively little is known about whether these traits repeatedly evolve through the same or different molecular and developmental mechanisms. The small freshwater fish,Astyanax mexicanus, provides an opportunity to investigate the repeated evolution of cave traits.A. mexicanusexists as two forms, a sighted, surface‐dwelling form and at least 29 populations of a blind, cave‐dwelling form that initially develops eyes that subsequently degenerate. We compared eye morphology and the expression of eye regulatory genes in developing surface fish and two independently evolved cavefish populations, Pachón and Molino. We found that many of the previously described molecular and morphological alterations that occur during eye development in Pachón cavefish are also found in Molino cavefish. However, for many of these traits, the Molino cavefish have a less severe phenotype than Pachón cavefish. Further, cave–cave hybrid fish have larger eyes and lenses during early development compared with fish from either parental population, suggesting that some different changes underlie eye loss in these two populations. Together, these data support the hypothesis that these two cavefish populations evolved eye loss independently, yet through some of the same developmental and molecular mechanisms.