Bivalves maintain repair when faced with chronically repeated mechanical stress.

Even though mollusks' capacity to repair shell damage is usually studied in response to a single event, their shells have to defend them against predatory and environmental threats throughout their potentially multi-decadal life. We measured whether and how mollusks respond to chronic mechanical stress. Once a week for 7 months, we compressed whole live California mussels (Mytilus californianus) for 15 cycles at ∼55% of their predicted one-time breaking force, a treatment known to cause fatigue damage in shells. We found mussels repaired their shells. Shells of experimentally stressed mussels were just as strong at the end of the experiment as those of control mussels that had not been experimentally loaded, and they were more heavily patched internally. Additionally, stressed shells differed in morphology; they were heavier and thicker at the end of the experiment than control shells but they had increased less in width, resulting in a flatter, less domed shape. Finally, the chronic mechanical stress and repair came at a cost, with stressed mussels having higher mortality and less soft tissue than the control group. Although associated with significant cost, mussels' ability to maintain repair in response to ongoing mechanical stress may be vital to their survival in harsh and predator-filled environments.

Bivalves rapidly repair shells damaged by fatigue and bolster strength.

Hard external armors have to defend against a lifetime of threats yet are traditionally understood by their ability to withstand a single attack. Survival of bivalve mollusks thus can depend on the ability to repair shell damage between encounters. We studied the capacity for repair in the intertidal mussel Mytilus californianus by compressing live mussels for 15 cycles at ∼79% of their predicted strength (critically fracturing 46% of shells), then allowing the survivors 0, 1, 2 or 4 weeks to repair. Immediately after fatigue loading, mussel shells were 20% weaker than control shells that had not experienced repetitive loading. However, mussels restored full shell strength within 1 week, and after 4 weeks shells that had experienced greater fatiguing forces were stronger than those repetitively loaded at lower forces. Microscopy supported the hypothesis that crack propagation is a mechanism of fatigue-caused weakening. However, the mechanism of repair was only partially explained, as epifluorescence microscopy of calcein staining for shell deposition showed that only half of the mussels that experienced repetitive loading had initiated direct repair via shell growth around fractures. Our findings document repair weeks to months faster than demonstrated in other mollusks. This rapid repair may be important for the mussels' success contending with predatory and environmental threats in the harsh environment of wave-swept rocky coasts, allowing them to address non-critical but weakening damage and to initiate plastic changes to shell strength. We highlight the significant insight gained by studying biological armors not as static structures but, instead, as dynamic systems that accumulate, repair and respond to damage.

Mechanical fatigue fractures bivalve shells.

Mollusk shells protect against diverse environmental and predatory physical threats, from one-time impacts to chronic, low-magnitude stresses. The effectiveness of shells as armor is often quantified with a test of shell strength: increasing force is applied until catastrophic fracture. This test does not capture the potential role of fatigue, a process by which chronic or repeated, low-magnitude forces weaken and break a structure. We quantified the strength and fatigue resistance of California mussel (Mytilus californianus) shells. Shells were fatigue tested until catastrophic failure by either loading a valve repeatedly to a set force (cyclic) or loading a valve under constant force (static). Valves fatigued under both cyclic and static loading, i.e. subcritical forces broke valves when applied repeatedly or for long durations. Stronger and more fatigue-resistant valves tended to be more massive, relatively wider and the right-hand valve. Furthermore, after accounting for the valves' predicted strength, fatigue resistance curves for cyclic and static loading did not differ, suggesting that fatigue fracture of mussels is more dependent on force duration than number of cycles. Contextualizing fatigue resistance with the forces mussels typically experience clarifies the range of threats for which fatigue becomes relevant. Some predators could rely on fatigue, and episodic events like large wave impacts or failed predation attempts could weaken shells across long time scales. Quantifying shell fatigue resistance when considering the ecology of shelled organisms or the evolution of shell form offers a perspective that accounts for the accumulating damage of a lifetime of threats, large and small.

Smashing mantis shrimp strategically impact shells.

Many predators fracture strong mollusk shells, requiring specialized weaponry and behaviors. The current shell fracture paradigm is based on jaw- and claw-based predators that slowly apply forces (high impulse, low peak force). However, predators also strike shells with transient intense impacts (low impulse, high peak force). Toward the goal of incorporating impact fracture strategies into the prevailing paradigm, we measured how mantis shrimp (Neogonodactylus bredini) impact snail shells, tested whether they strike shells in different locations depending on prey shape (Nerita spp., Cenchritis muricatus, Cerithium spp.) and deployed a physical model (Ninjabot) to test the effectiveness of strike locations. We found that, contrary to their formidable reputation, mantis shrimp struck shells tens to hundreds of times while targeting distinct shell locations. They consistently struck the aperture of globular shells and changed from the aperture to the apex of high-spired shells. Ninjabot tests revealed that mantis shrimp avoid strike locations that cause little damage and that reaching the threshold for eating soft tissue is increasingly difficult as fracture progresses. Their ballistic strategy requires feed-forward control, relying on extensive pre-strike set-up, unlike jaw- and claw-based strategies that can use real-time neural feedback when crushing. However, alongside this pre-processing cost to impact fracture comes the ability to circumvent gape limits and thus process larger prey. In sum, mantis shrimp target specific shell regions, alter their strategy depending on shell shape, and present a model system for studying the physics and materials of impact fracture in the context of the rich evolutionary history of predator-prey interactions.

Mechanical properties of sediment determine burrowing success and influence distribution of two lugworm species.

We apply new perspectives on how organisms burrow by examining the association of in situ variation in sediment mechanical properties with burrowing ability and species distribution of two sympatric lugworms, Abarenicola pacifica and Abarenicola claparedi We quantified the sediment's resistance to penetration and its grain size distribution at sites inhabited by each species. Abarenicola pacifica individuals were found in significantly harder to penetrate, more heterogeneous sediments. We compared worm burrowing ability using reciprocal transplant experiments. Worms from firmer sediments, A. pacifica, were able to make successful steep burrows in sediments characteristic of either species. In contrast, A. claparedi individuals often failed to complete successful burrows in the firmer A. pacifica sediment. To examine how morphological differences could explain these patterns, we compared body wall musculature and measured how well individuals support their own bodies when draped over a cantilever. Lugworms from the firmer sediment had thicker body wall musculature and held their bodies more rigidly than did worms from softer sediments. Additionally, we observed subtle differences in the papillae on the proboscises' surfaces, which could affect worm-sediment interactions, but we found no differences in the chaetae of the two species. Abarenicola claparedi produced more mucus, which could be important in shoring up burrow walls in their shifting, sandy habitat. This study presents the first example of using field-based experiments to determine how sediment mechanical properties and worm burrowing ability could act to determine organismal distribution. Our findings have broader ecological implications because of the role of lugworms as ecosystem engineers.

Kinetic study of the helix to coil dark reaction of poly(spiropyran-L-glutamate).

An investigation of kinetics of the helix to coil dark reaction of light adapted poly(spiropyran-L-glutamic acid) (PSLG) dissolved in hexafluoroisopropanol was performed. The reaction was associated with the spiropyran (SP) to merocyanine (MC) ring opening. The ring opening reaction monitored with UV/VIS spectroscopy showed first order kinetics. Chromophore and polypeptide backbone circular dichroism data fit to an expression consistent with a single intermediate series mechanism. By FTIR, we monitored the polypeptide alpha-helix amide I, the MC chromophore--C = C--stretch and the protonated unmodified carboxylate C = O stretch bands. During the first step of the series mechanism, changes in the hydrogen bonding of the unmodified carboxylate groups occurred, suggesting breakup of polypeptide aggregates. The second step of the proposed series mechanism was dominated by the helix to coil transition and the ring opening of SP to MC. The CD spectrum of MC in the dark adapted PSLG was red shifted and had a narrower bandwidth than the UV/VIS spectrum. The kinetic and spectroscopic data suggested that a fraction (population I) of the MC chromophores experienced optical activity induced by the chiral polypeptide environment, while the remainder of the MC chromophores (population II) were solvated and enantiomeric.

Investigation of light-induced conformation changes in spiropyran-modified succinylated poly(L-lysine).

To determine the maximum range of coupling between side-chain photochromism and polypeptide conformation change, we modified the carboxylate side chains of succinylated poly(L-lysine) with a spiropyran to form polypeptide I. The extent of modification was determined to be 35.5%. The spacer group length between the polypeptide alpha-carbon and the dye was 12 atoms, providing minimum polypeptide-dye interaction. Conformation changes were monitored by circular dichroism as a function of light adaptation and solvent composition (hexafluoroisopropanol [HFIP] vs trifluoroethanol [TFE]). Under all solvent compositions, the dark-adapted dye was in the merocyanine form. Light adaptation by visible light converted the dye to the spiropyran form. When dissolved in TFE, I adopted a helical conformation insensitive to light adaptation. With increasing percentage HFIP, a solvent-induced helix-to-coil transition was observed around 80% (vol/vol) HFIP. At 100% HFIP, both light- and dark-adapted forms of I were in the coil state. Near the midpoint of the solvent-induced helix-to-coil transition, light adaptation caused conformation changes. Applying helix-to-coil transition theory, we measured a statistically significant difference in coil segment-HFIP binding constant for light- vs dark-adapted solutions (6.38 +/- 0.03 M-1 vs 6.56 +/- 0.03 M-1), but not for the nucleation parameter sigma (1.2 +/- 0.4 10(-3) vs 1.3 +/- 0.3 x 10(-3). The small binding constant difference translated to a light-induced binding energy difference of 17 cal/mol/monomer. Near the midpoint of the helix-to-coil transition, collective interactions between monomer units made possible the translation of a small energy difference (less than RT) into large macromolecular conformation changes.(ABSTRACT TRUNCATED AT 250 WORDS)

Optical limiting in solutions of diphenyl polyenes.

The optical-limiting behavior of a series of trans-alpha, omega-diphenyl polyene compounds was observed in solutions of chloroform. The influence of planarity, the substitution of donor and acceptor groups, and the extent of pi-electron delocalization on the nonlinear thresholds of the diphenyl polyenes in an optical-limiting geometry were examined. A saturation effect of optical-limiting nonlinear thresholds, consistent with the theoretical work, was observed at 10-11 pi-electron bonds. The temporal profiles of the transmitted laser pulses and the power dependence of the nonlinear thresholds as a function of spot size and wavelength were examined. These examinations led to the conclusion that the predominant nonlinear mechanism was quasi-steady-state self-focusing. Nonlinear thresholds an order of magnitude lower, and thus effective n(2) and X(3) values an order of magnitude higher, than the well-known self-focusing medium CS(2) were observed. Our studies demonstrate that this series of polyenes consists of efficient broadband nonresonant optical-limiting materials.

Holographic grating formation in poly(spiropyran I-glutamate).

An argon-ion laser interferometer (0.3638 or 0.488 microm) wrote photoreversible holographic grating (fringe spacing 0.39-100 microm) on spin-cast thin films (5 microm thick) of poly(spiropyran l-glutamate).