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Barnacles adhere by producing a mixture of cement proteins (CPs) that organize into a permanently bonded layer displayed as nanoscale fibers. These cement proteins share no homology with any other marine adhesives, and a common sequence-basis that defines how nanostructures function as adhesives remains undiscovered. Here we demonstrate that a significant unidentified portion of acorn barnacle cement is comprised of low complexity proteins; they are organized into repetitive sequence blocks and found to maintain homology to silk motifs. Proteomic analysis of aggregate bands from PAGE gels reveal an abundance of Gly/Ala/Ser/Thr repeats exemplified by a prominent, previously unidentified, 43 kDa protein in the solubilized adhesive. Low complexity regions found throughout the cement proteome, as well as multiple lysyl oxidases and peroxidases, establish homology with silk-associated materials such as fibroin, silk gum sericin, and pyriform spidroins from spider silk. Distinct primary structures defined by homologous domains shed light on how barnacles use low complexity in nanofibers to enable adhesion, and serves as a starting point for unraveling the molecular architecture of a robust and unique class of adhesive nanostructures.

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The development of non-toxic or non-polluting antifouling additives that can be formulated in practical coatings requires assays involving target organisms. Assays that test both for the effective and toxic concentrations of active compounds are useful. It is also desirable if the assay can provide information regarding the performance that can be expected if the compounds are incorporated into different matrices. Described here are the simple laboratory assays that have been developed using the barnacle, Balanus amphitrite, a common fouling organism found throughout temperate and tropical seas. One of the assays depends on synchronous year-round mass culture, the procedure for which is described, of cypris larvae. The laboratory assays provide quantitative estimates of toxicity and settlement inhibition of barnacles. The methods described provide an excellent system for the use of barnacles to study the interaction of the test compounds and surfaces.

Biologically active extracts of the Caribbean sponge Agelas conifera have yielded, in exhaustive studies, the diacetate salts of seven new bromopyrroles (1,(3)(4)(5)(6)(7)(8), as well as that of the known debromooroidin dimer sceptrin (2). These compounds were found to be antiviral and antibacterial and were active in barnacle settlement and biochemical prophage induction assays. The structures assigned were based on spectroscopic comparisons to sceptrin and two-dimensional NMR data. Synthetic bromopyrroles were used to verify bromine substitution patterns. The oxysceptrins (4,5) are characterized by their aminoimidazolinone group, the ageliferins (6-8) by a unique cyclohexene-based skeleton.

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Increased levels of atmospheric CO 2 are anticipated to cause decreased seawater pH. Despite the fact that calcified marine invertebrates are particularly susceptible to acidification, barnacles have received little attention. We examined larval condition, cyprid size, cyprid attachment and metamorphosis, juvenile to adult growth, shell calcium carbonate content, and shell resistance to dislodgement and penetration in the barnacle Amphibalanus amphitrite reared from nauplii in either ambient pH 8.2 seawater or under CO 2 -driven acidification of seawater down to a pH of 7.4. There were no effects of reduced pH on larval condition, cyprid size, cyprid attachment and metamorphosis, juvenile to adult growth, or egg production. Nonetheless, barnacles exposed to pH 7.4 seawater displayed a trend of larger basal shell diameters during growth, suggestive of compensatory calcification. Furthermore, greater force was required to cause shell breakage of adults raised at pH 7.4, indicating that the lower, active growth regions of the wall shells had become more heavily calcified. Ash contents (predominately calcium carbonate) of basal shell plates confirmed that increased calcification had occurred in shells of individuals reared at pH 7.4. Despite enhanced calcification, penetrometry revealed that the central shell wall plates required significantly less force to penetrate than those of individuals raised at pH 8.2. Thus, dissolution rapidly weakens wall shells as they grow. The ramifications of our observations at the population level are important, as barnacles with weakened wall shells are more vulnerable to predators.

No abstract