Chemosensory Exploitation and Predator-Prey Arms Races

Thousands of armed predatory species, distributed widely across the metazoan tree-of-life, consume only hard-shell or exoskeleton-bearing organisms (called “durophagy”). Prey armor clearly has evolved in response to selection by predators, but there is little evidence of the contrary, counter-adaptation by predators. Evolved consumer responses to prey, in general, might be more readily expressed in ways other than morphological traits, including via sensory cues. Here, we explored the chemosensory basis for durophagy in a model predator-prey system, and identified intimate associations between durophagous predators and their shelled prey. Barnacles (Balanus glandula and Semibalanus cariosus) bear hard shells and secrete, respectively, a 199 or 201 kDa glycoprotein ortholog (named “MULTIFUNCin”), with expression limited to the body armor (epidermis, cuticle, and live shell). To test for effects of MULTIFUNCin on predators, we constructed faux prey to mimic meaningful physical and chemical characteristics of live barnacles. In separate experiments, each consumer species was presented MULTIFUNCin, purified from either B. glandula or S. cariosus, at a typical armor concentration. All six predatory species (sea star, Pisaster ochraceus; whelks, Acanthinucella spirata, Nucella emarginata, N. ostrina, N. canaliculata, and N. lamellosa) attacked and ate MULTIFUNCin-infused faux prey significantly more than controls. Akin to barnacles, secretion of glycoprotein-rich extracellular matrices is common among armored prey species—from marine sponges to terrestrial vertebrates. Our results, therefore, suggest that chemosensory exploitation of glycoproteins could be widespread, with notable consequences for life on land and in the sea.

Effects of intertidal position on metabolism and behavior in the acorn barnacle, Balanus glandula

The intertidal zone is characterized by persistent, tidally-driven fluctuations in both abiotic (e.g., temperature, [O2], salinity) and biotic (e.g., food availability, predation) factors, which make this a physiologically challenging habitat for resident organisms. The relative magnitude and degree of variability of environmental stress differs between intertidal zones, with the most extreme physiological stress often being experienced by organisms in the high intertidal. Given that so many of the constantly shifting parameters in this habitat are primary drivers of metabolic rate (e.g., temperature, [O2], food availability), we hypothesized that sessile conspecifics residing in different tidal zones would exhibit distinct ‘metabolic phenotypes,’ a term we use to collectively describe the organisms’ baseline metabolic performance and capacity. To investigate this hypothesis, we collected acorn barnacles (Balanus glandula) from low, mid, and high intertidal positions in San Luis Obispo Bay, CA and measured a suite of biochemical (whole-animal citrate synthase (CS) and lactate dehydrogenase (LDH) activity, aerial [D-lactate]), physiological (O2 consumption rates), morphological (body size) and behavioral (e.g., cirri beat frequency, % time operculum open) indices of metabolism. We found tidal zone-dependent differences in B. glandula metabolism that primarily related to anaerobic capacity, cirral activity patterns and body size. Barnacles from the low intertidal tended to have a greater capacity for anaerobic metabolism (i.e., increased LDH activity, increased baseline [D-lactate]), have reduced cirral beating activity—and presumably reduced feeding—when submerged, and be smaller in size compared to conspecifics in the high intertidal. We did not, however, see any D-lactate accumulation in barnacles from any tidal height throughout the 96 h of air exposure. This trend indicates that the enhanced capacity of low intertidal barnacles for anaerobic metabolism may have evolved to support metabolism during more prolonged episodes of emersion or during events other than emersion (e.g., coastal hypoxia, predation). There were also no significant differences in CS activity or baseline oxygen consumption rates (in air or seawater at 14˚C) across tidal heights, which implies that aerobic metabolic capacity may not be as sensitive to tidal position as anaerobic processes. Understanding how individuals occupying different shore heights differ in their metabolic capacity becomes increasingly interesting in the context of global climate change, given that the intertidal zone is predicted to experience even greater extremes in abiotic stress.

Swimming kinematics of cyprids of the barnacle Balanus glandula

Larvae of barnacles typically pass through naupliar and cyprid planktonic stages before settlement and metamorphosis. As the final larval stage, cyprids swim much faster than nauplii and in turbulent fluid environments with high shears as they seek habitat. Cyprids swim with six pairs of reciprocating thoracic appendages and use two anterior antennules during settlement. Our understanding of how thoracic appendages generate movement is limited due to short stroke intervals (∼5 ms) that impede observations of the shape and trajectory of appendages. Here, we used high speed videography to observe both free-swimming and tethered cyprids of the intertidal acorn barnacle Balanus glandula to produce a comprehensive description of thoracic appendage swimming kinematics. Cyprids used a drag-based method of swimming: their six pairs of thoracic appendages moved through metachronal power strokes and synchronous recovery strokes similar to the thoracopod motions in calanoid copepods during escape swimming. During the power stroke, plumose setae on each appendage pair spread laterally into a high surface area and high drag paddle composed of a meshwork of fused setules. This interconnected setal array collapsed into a low surface area and low drag shape during the recovery stroke. These effective swimming appendages allowed cyprids to move upwards at an average speed of 1.4 cm s−1 (about 25 body lengths s−1) with an average beat frequency of 16 beats s−1, and reach an instantaneous velocity of up to 6 cm s−1. Beat frequency of the thoracic appendages was significantly associated with speed, with higher beat frequencies indicating faster swimming speed. At their average speed, cyprids moved at the intermediate Reynolds number of ∼10, in which both viscous and inertial forces affected movement. Cyprids could alter swimming direction by sweeping the posterior-most appendage pair to one side and beating the remaining thoracic appendages synchronously through the power stroke with greater motion on the outside of their turn. These results greatly enhance our understanding both of cyprid motility and how small planktonic organisms can use swimming appendages with fused setule arrays to reach high swimming speeds and affect directional changes.

Transition in Population Dynamics of the Intertidal Barnacle Balanus glandula after Invasion: Causes and Consequences of Change in Larval Supply

To elucidate how the population dynamics of the acorn barnacle Balanus glandula transitioned after its invasion in 2000 along the Pacific coast of Japan, a population census was conducted from 2004 to 2014 at five shores along 49 km of coastline 144–193 km east outside of the invasion front. Survey areas at each shore consisted of five paired plots (cleared recruitment plots and control plots). Larval recruitment was first detected in 2004 but benthic individuals were not detected until 2 years later. The abundance and occurrence of B. glandula increased until around 2010; abundance then decreased but occurrence remained high (70%) until 2014, suggesting that the metapopulation of this barnacle approached a maximum around 2011. From 2011, the population dynamics of B. glandula changed considerably at two contrasting spatial scales: at a regional scale, the dependency of the number of larvae on stock size decreased, whereas at a local scale, the relative contribution of larval supply as a determinant of local population dynamics decreased. These findings suggest that the major driving force of population dynamics of the introduced barnacle changed in just a few years after invasion; therefore, population census data from just after an invasion, including larval recruitment monitoring just outside the invasion front, is essential to understanding invasion dynamics by sessile marine organisms.

D-lactate production in the acorn barnacle Balanus glandula (Darwin, 1854) (Cirripedia: Balanidae) under emersion stress

Anaerobic metabolism is an important response to stress in many organisms. Intertidal species often face heat stress during low tide. Balanus glandula (Darwin, 1854) is a high-shore intertidal barnacle common to the Pacific that experiences prolonged low-tide air exposure. It is not known whether B. glandula uses anaerobic metabolism during emersion, or if its use varies by latitude. We measured low tide D-lactate production in two US west coast populations of B. glandula separated by 14 degrees of latitude. We exposed barnacles to seven low-tide air temperatures (10, 15, 20, 25, 30, 35, and 38 °C) for which aerobic respiration has been previously measured. Our northern population of B. glandula increased D-lactate production at high air temperatures where aerobic metabolic depression is known to occur, indicating sublethal stress. In contrast, our southern population showed little increase in D-lactate over the same temperature range, coincident with high aerobic respiration across those temperatures. In a second experiment, we quantified D-lactate at 1, 2, and 6 hours post-emersion for northern B. glandula exposed to either a 10 or 38 °C low tide, to measure their potential lactate usage. While D-lactate was elevated at 38 °C compared to the 10 °C control immediately following low tide exposure, it dropped to control levels, and was likely excreted, within 1 hour of re-immersion. Our results suggest that the low latitude population of B. glandula may be more resilient to climate change than its high latitude counterpart in the absence of adaptation, which has strong implications for species distribution.

Limpet disturbance effects on barnacle recruitment are related to recruitment intensity but not recruit size

Intertidal limpets are important grazers along rocky coastlines worldwide that not only control algae but also influence invertebrates such as common barnacles. For instance, grazing limpets ingest settling barnacle cyprid larvae (hereafter cyprids) and push cyprids and barnacle recruits off the substrate. Such limpet disturbance effects (LDEs) can limit barnacle recruitment, a key demographic variable affecting barnacle population establishment and persistence. In this study, we examined limpet (Lottia cassis) disturbance to barnacle (Chthamalus dalli, Balanus glandula) recruitment on the Pacific coast of Hokkaido, Japan, as information on limpet-barnacle interactions from this region is missing. We investigated, for the first time, whether barnacle size and recruitment intensity influence LDEs on barnacle recruitment. Small barnacles may be less susceptible to LDEs than larger barnacles, because small size may reduce the propbability of limpet disturbance. Moreover, recruitment intensity can influence LDEs, as high recruitment can compensate for LDEs on barnacle recruitment density. In Hokkaido, C. dalli cyprids are smaller than B. glandula cyprids, and C. dalli recruitment is higher than B. glandula recruitment. Thus, we hypothesized that LDEs on C. dalli recruitment would be weaker than those on B. glandula recruitment. To test our hypothesis, we conducted a field experiment during which we manipulated limpet presence/absence on the interior surfaces of ring-shaped cages. After four weeks, we measured barnacle recruitment and recruit size on the interior surfaces of the cages and found negative LDEs on C. dalli and B. glandula recruitment and recruit size. As hypothesized, the LDEs on C. dalli recruitment were weaker than the LDEs on B. glandula recruitment. Additionally, C. dalli recruits were smaller than B. glandula recruits. However, the LDEs on C. dalli recruit size were as strong as the LDEs on B. glandula recruit size, indicating that the smaller C. dalli recruits are not less susceptible to LDEs than B. glandula recruits. Since C. dalli recruitment was higher than B. glandula recruitment, we propose that the higher C. dalli recruitment compensated for the LDEs on C. dalli recruitment. Our findings indicate that the detected differences in LDEs on barnacle recruitment are related to barnacle recruitment intensity but not recruit size.

Diversity and distribution of nearshore barnacle cyprids in southern California through the 2015–16 El Niño

Abundance, species diversity, and horizontal distributions of barnacle cyprids offshore of La Jolla, southern California were described from May 2014 to August 2016 to determine how the nearshore barnacle larval assemblage changed before, during, and after the 2015–16 El Niño. The entire water column was sampled at five stations located within one km of shore with water depths of 4, 6, 8, 10, and 12 m during 33 cruises that encompassed the time when El Niño conditions impacted the area. Nearshore temperature and thermal stratification was concurrently measured using a CTD. Six identified cyprid species, including Chthamalus fissus, Pollicipes polymerus, Megabalanus rosa, Tetraclita rubescens, Balanus glandula, and B. trigonus, along with four unknown species, were collected in our samples. DNA barcoding was used to confirm identifications in a subset of the larvae. C. fissus was more than eight times more abundant than any other species, and while abundance varied by species, cyprid density was highest for all species except for M. rosa before and after the El Niño event, and lower during the environmental disturbance. There were significant differences in cross-shore distributions among cyprid species, with some located farther offshore than others, along with variability in cross-shore distributions by season. C. fissus cyprids were closest to shore during spring-summer cruises when waters were the most thermally stratified, which supports previous findings that C. fissus cyprids are constrained nearshore when thermal stratification is high. Relative species proportions varied throughout the study, but there was no obvious change in species assemblage or richness associated with El Niño. We speculate that barnacle cyprid species diversity did not increase at our study site during the 2015–16 El Niño, as it has in other areas during previous El Niño Southern Oscillation events, due to the lack of anomalous northward flow throughout the 2015–16 event.