This is a study of the foraging behaviour of Carcinus maenas on Mytilus edulis and of Thalamita danae on Perna viridis. Particular attention is given to differences arising in foraging behaviour as a result of intraspecific prey heterogeneity and experimental protocol. Intersite and temporal differences in the population density, shell morphology, biomass and byssal attachment strength of Mytilus edulis were found. Byssal attachment strength and shell strength were highly variable amongst individuals of a similar size. Carcinus maenas is strongly heterochelous. Intraspecific differences in the chelal mechanics, but not in the chelal geometry, were recorded; major chelae of large male crabs were significantly stronger than the major chelae of females and small males. Stomach content analyses showed that Carcinus maenas has a broad diet in which Mytilus edulis forms an important component. Intersite differences inMytilus edulis shell morphology altered the foraging behaviour of Carcinus maenas, and intersite and temporal variations in mussel flesh weight altered the prey value curves. Both C. maenas and Thalamita danae were highly prey size-selective when foraging on groups of different sized mussels, the size of prey most vulnerable to predation altering with the size composition of the group. The handling times of mussels for both species of crab were reduced when mussels were presented as part of a group as compared to when mussels were presented singly. For Carcinus maenas the reduced handling times resulted from the less extensive gleaning of mussel shells whilst for Thalamita danae reduced handling times appeared to result from the greater use of a more time efficient opening technique. When Carcinus maenas were presented with mussels of differing attachment strengths, crabs selected more weakly attached mussels over those with a more firm and rigid attachment. This selection did not appear to be based on prey value or prey length but rather on the resulting slight movement of weakly attached mussels whenever these were touched by a foraging crab.

The following research deals with the response of the small shore crabs, Hemigrapsus oreqonesis and Hemigrapsus nudus to paralytic shellfish toxins (PST). These shore crabs were shown to develop a remarkable seasonal resistance to administered saxitoxin (STX). No similar change in sensitivity was found after administration of tetrodotoxin (TTX), another marine neurotoxin with similar actions to the PST. Resistance to STX in the small shore crabs was linked to the presence of PST in the viscera, and this in turn was related to the presence of toxic dinoflagellate blooms in the area. Furthermore, this research provides, for the first time, evidence of a protein component (MW 145,000 daltons) which appears to be associated with acquired resistance to PST in the shore crab. In addition, this protein component was shown to appear in sensitive crab extracts after the administration of low doses of saxitoxin and tetrodotoxin in vivo. / Land and Food Systems, Faculty of / Graduate

Hydrostatic pressure sensing is used by aquatic animals to estimate depth and to synchronize behaviour with the tides. Pressure sensors examined to date depend on the compression of a fluid, making them susceptible to temperature change. Despite this, the effects of temperature on pressure sensing have not so far been researched. This thesis examined the effects of temperature on the afferent firing rate of thread hair pressure receptors in the crab Carcinus maenas. It tested the hypothesis that for sensors responding positively to rising pressure, an increase in temperature should counteract any contraction in volume, thereby leading to a weaker neural response; vice versa, a stronger response was predicted for a temperature decrease. A novel system was developed which allowed the simultaneous application of sinusoidal pressure change from 150 to 350 mbar and cycles of temperature, typically spanning between 16 and 22 °C, while extracellular recordings were made en passant from afferent nerves innervating statocyst thread hairs. Motor programmes were designed to stimulate thread hairs sensitive to clockwise and anticlockwise rotation through continuous or interrupted bouts of oscillation within the horizontal plane. During constant pressure, raising temperature led to increases in spike frequency and amplitude, with Q10s between 3 and 5 for spike frequency but less than 2 for amplitude. Cooling caused the elimination or decrease of pressure responses while increasing them on heating, thereby contradicting the aforementioned hypothesis of temperature mimicking pressure change. Changes in the timing of pressure responses were also observed. In preparations which were non-responsive to pressure change, temperature change led to bursts in firing at the peaks or troughs of pressure cycles in 28 of 42 preparations, with evidence for recruitment of formerly silent units. The results could not be fully explained under the existing model for pressure sensing by thread hairs, highlighting the necessity for further anatomical studies.

Thesis (M.S.)--Oregon State University, 2002. / Typescript (photocopy). Includes bibliographical references (leaves 63-105). Also available via the World Wide Web.

Theses (Ph.D.)--University of New Hampshire (Dept. of Zoology), 2007. / Includes bibliographical references. Also available online.

Invasion physiology is an emerging field that endeavors to understand the influence of physiological traits on the establishment of non-native species in novel environments. The invasive European green crab,Carcinus maenas, is one of the world's most successful aquatic invaders, and is currently distributed across temperate marine ecosystems globally. The work presented here explored the thermal physiology of this species, and has highlighted several physiological traits that have likely influenced establishment success. Intraspecific comparisons of crabs sampled from the northern and southern edges of their recipient, or invaded range on the west coast of North America have identified both organismal and cellular physiological difference with respect to upper and lower thermal tolerances. Crabs sampled from British Columbia, Canada (BC) had a significantly lower mean upper thermal tolerance threshold and heat shock protein synthesis, Hsp70, compared to their warm acclimated conspecifics sampled from California (CA). These differential physiologic responses may be rooted in the disparate natural thermal habitats that each population occupies within their respective environments. The ability of this species to extend its current range limits was also investigated. Range expansion to the south has been limited, and is likely restricted by this species lack of adaptation to warmer temperatures. Because range expansion has been chiefly northward, characterizing this species' response to cold stress can identify whether colder temperatures poleward may limit further range expansion. Cold tolerance capacity was determined in the laboratory, and crabs sampled from Vancouver Island, British Columbia were able to withstand the over-wintering thermal regime that occurs in Sitka, Alaska, a site that is currently beyond the range limits of this species. Furthermore, intraspecific assessments found that the cold acclimated BC population exposed to cold shock significantly down regulated protein levels of cyclin D1, cell cycle modulator. Distinct differences in carapace width (CW) were detected along the thermal gradient present in the green crabs' range. This variation in body size was utilized to the test the temperature size rule hypothesis for ectotherms. Simply stated, the temperature size rule is the tendency for ectotherms to develop slower but mature to a larger body sizes at cooler temperatures. The results supported this hypothesis as crabs sampled from the warm portion of the range were found to be smaller than crabs sampled from the colder portion of the range. This pattern was detected along the native range as well. Differences in body size have the potential to influence the scope of invasion; larger individuals are generally more fecund and longer lived, which can increase both the intensity and frequency of larval dispersal that could further propel range expansion. The physiologic properties that the green crab possesses which may influence invasion success were examined using peer-reviewed literature with the aim of determining if these physiological traits confer invasion success across taxa. This analysis tested four hypotheses: 1) Broad geographic temperature tolerances (thermal width) confer a higher upper thermal tolerance threshold when comparing invasive and native species. 2) The upper thermal extreme experienced in nature is correlated with upper thermal tolerance threshold. 3) Protein chaperone expression, a cellular mechanism underlying thermal tolerance threshold, is greater in invasive organisms than in native ones. 4) Acclimation to higher temperatures can promote a greater range of thermal tolerance for invasives compared to natives. These preliminary results generally support the four stated hypotheses, and provide a solid foundation for further studies to explore and identify physiologic traits that facilitate invasion success. Overall, these studies investigated the thermal physiology ofCarcinus maenasfrom an invasive metapopulation and have brought about significant advances in our understanding of what physiologic traits correlate to invasion success in this species. In addition, the data presented here can aid resource managers in identifying habitats, based on thermal tolerance measurements that fit the criteria for invasion. Understanding how invasive organisms vary with respect to thermal tolerance can aid our understanding the patterns and processes of species invasions.

Over the last decades the crustacean aquaculture sector has been steadily growing, in order to meet global demands for its products. A major hurdle for further growth of the industry is the prevalence of viral disease epidemics that are facilitated by the intense culture conditions. A devastating virus impacting on the sector is the White Spot Syndrome Virus (WSSV), responsible for over US $10 billion in losses in shrimp production and trade. The Pathogenicity of WSSV is high, reaching 100 % mortality within 3-10 days in penaeid shrimps. In contrast, the European shore crab Carcinus maenas has been shown to be relatively resistant to WSSV. Uncovering the basis of this resistance could help inform on the development of strategies to mitigate the WSSV threat. C. maenas has been used widely in studies on ecotoxicology and host-pathogen interactions. However, like most aquatic crustaceans, the genomic resources available for this species are limited, impairing experimentation. Therefore, to facilitate interpretations of the exposure studies, we first produced a C. maenas transcriptome and genome scaffold assembly. We also produced a transcriptome for the European lobster (Homarus gammarus), an ecologically and commercially important crustacean species in United Kingdom waters, for use in comparing WSSV responses in this, a susceptible species, and C. maenas. For the C. maenas transcriptome assembly we isolated and pooled RNA from twelve different tissues and sequenced RNA on an Illumina HiSeq 2500 platform. After de novo assembly a transcriptome encompassing 212,427 transcripts was produced. Similar, the H. gammarus transcriptome was based on RNA from nine tissues and contained 106,498 transcripts. The transcripts were filtered and annotated using a variety of tools (including BLAST, MEGAN and RSEM) and databases (including GenBank, Gene Ontology and KEGG). The annotation rate for transcripts in both transcriptomes was around 20-25 % which appears to be common for aquatic crustacean species, as a result of the lack of well annotated gene sequences for this clade. Since it is likely that the host immune system would play an important role in WSSV infection we characterized the IMD, JAK/STAT, Toll-like receptor and other innate immune system pathways. We found a strong overlap between the immune system pathways in C. maenas and H. gammarus. In addition we investigated the sequence diversity of known WSSV interacting proteins amongst susceptible penaeid shrimp/lobster and the more resistant C. maenas. There were differences in viral receptor sequences, like Rab7, that correlate with a less efficient infection by WSSV. To produce the genome scaffold assembly for C. maenas we isolated DNA from muscle tissue and produced both paired-end and mate pair libraries for processing on the Illumina HiSeq 2500 platform. A de novo draft genome assembly consisting of 338,980 scaffolds and covering 362 Mb (36 % of estimated genome size) was produced, using SOAP-denovo2 coupled with the BESST scaffolding system. The generated assembly was highly fragmented due to the presence of repetitive areas in the C. maenas genome. Using a combination of ab initio predictors, RNA-sequencing data from the transcriptome datasets and curated C. maenas sequences we produced a model encompassing 10,355 genes. The gene model for C. maenas Dscam, a gene potentially involved in (pan)crustacean immune memory, was investigated in greater detail as manual curation can improve on the results of ab initio predictors. The scaffold containing C. maenas Dscam was fragmented, thus only contained the latter exons of the gene. The assembled draft genome and transcriptomes for C. maenas and H. gammarus are valuable molecular resources for studies involving these and other aquatic crustacean species. To uncover the basis of their resistance to WSSV, we infected C. maenas with WSSV and measured mRNA and miRNA expression for 7 time points spread over a period of 28 days, using RNA-Seq and miRNA-Seq. The resistance of C. maenas to WSSV infection was confirmed by the fact that no mortalities occurred. In these animals replicating WSSV was latent and detected only after 7 days, and this occurred in five of out 28 infected crabs only. Differential expression of transcripts and miRNAs were identified for each time point. In the first 12 hours post exposure we observed decreased expression of important regulators in endocytosis. Since it is established that WSSV enters the host cells through endocytosis and that interactions between the viral protein VP28 and Rab7 are important in successful infection, it is likely that changes in this process could impact WSSV infection success. Additionally we observed an increased expression of transcripts involved in RNA interference pathways across many time points, indicating a longer term response to initial viral exposure. miRNA sequencing showed several miRNAs that were differentially expressed. The most striking finding was a novel C. maenas miRNA that we found to be significantly downregulated in every WSSV infected individual, suggesting that it may play an important role in mediating the response of the host to the virus. In silico target prediction pointed to the involvement of this miRNA in endocytosis regulation. Taken together we hypothesize that C. maenas resistance to WSSV involves obstruction of viral entry by endocytosis, a process probably regulated through miRNAs, resulting in inefficient uptake of virions.

Molting in arthropods is a complex process governed by regulatory mechanisms that have evolved and adapted over millennia to allow these animals to grow, despite being confined by a hardened exoskeleton. We isolated the molt-regulating Y-organs (YO) from the common shore crab Carcinus maenas at molt stages B, C1-3, C4, and D0 to assess how changes in protein abundances might underline the unique physiology of each of these stages. We found that changes in protein abundance were most notable in the postmolt stages (B and C1-3), where an increase in energy metabolism and the reactive oxygen species stress (ROS) response proteins was observed. An increase in triosephosphate isomerase and transketolase suggest that the postmolt YO is participating in triglycerides storage and is also actively recycling excess ribose sugars manufactured during the YO’s previously activated state. We also propose as mechanism through which ROS-induced release of cyclophilin A may contribute to YO atrophy during postmolt through the remodeling of structural proteins such as collagen. We support the standing observation of YO atrophy during postmolt by drawing attention to hemolymph protein abundances, especially those of cryptocyanin isoforms, which dropped precipitously in intermolt (C4) and remained at low abundances into early premolt (D0). Finally, though our evidence is preliminary, we propose that future investigations into the YO proteome address the significance of the protein glutamate dehydrogenase. Glutamate dehydrogenase, a key enzyme involved in the formation of glutamate, represents a potential nutrient-sensing checkpoint that might be involved in YO activation. Historically, most attention has gone to the acute molt stages, where signaling mechanisms involved in the activation of the YO have been the focus. Here, we present data suggesting that other regulatory mechanism may be governing the atrophy the postmolt YO. A better understanding of crustacean physiology has the potential to benefit ecosystems and economies worldwide.

Lethal biotic interactions strongly influence the potential for aquatic non-native species to establish and endure in habitats to which they are introduced. Predators in the recipient area, including native and previously established non-native predators, can prevent establishment, limit habitat use, and reduce abundance of non-native species. Management efforts by humans using methods designed to cause mass mortality (e.g., trapping, biocide applications) can reduce or eradicate non-native populations. However, the impacts of predator and human induced mortality may be mitigated by the behavior or population-level responses of a given non-native species. My dissertation examined the responses of non-native aquatic species to the risk of predation by novel (i.e., no previous exposure) predators in the recipient community and indicators of potential compensatory responses by non-native populations to increased mortality resulting from removal efforts. My dissertation addresses four primary questions. 1) Can first generation, naïve invaders recognize and defend against predators found within the region of invasion through the expression of inducible defenses? 2) Can the overcompensatory potential of a population be predicted through examinations of intraspecific interactions of individuals from the population? 3) What is the relationship between removal effort outcome (i.e., successful or unsuccessful reduction of the target population) and compensatory population responses? 4) Is there a relationship between characteristics of removal efforts that are typically available to managers (e.g., target area size, target area connectivity, removal methodology) and compensatory population responses that could indicate the relative likelihood of compensation resulting from removal efforts? An invading species should be more likely to establish if it can successfully identify and defend against predators in the recipient range, such as through the expression of inducible defenses. Inducible defenses are behavioral or physiological changes that reduce an organism's susceptibility to predation. Through a series of laboratory experiments, I tested whether inducible defenses, in the form of increased burrowing depth, may have benefited the early stage of invasion of Nuttallia obscurata (purple varnish clam), an established Northeast Pacific invader. Specimens of N. obscurata were collected from introduced populations in the Northeast Pacific and from a native population in Japan. The clams were exposed to chemical and physical cues from Northeast Pacific crab predators, including the native Metacarcinus magister (Dungeness crab), an abundant and frequent predator of N. obscurata. While introduced N. obscurata increased their burrowing depth in the physical presence of M. magister, clams collected from their native range showed no such response. This lack of increased burrowing depth by naïve clams in response to a predator native to the newly invaded range, but a significant increase in depth for clams from populations established in the range suggests that while inducible defenses likely did not contribute to the initial establishment of N. obscurata in the Northeast Pacific, they may contribute to their continued persistence and expansion in their introduced range. Some efforts to reduce invasive populations have paradoxically led to population increases. This phenomenon, referred to as overcompensation, occurs when strong negative density-dependent interactions are reduced through increased mortality within a population, resulting in an increase in the population's recruitment rate sufficient to increase the population's overall abundance. Increases in a population's recruitment rate can result from reduced cannibalism of juveniles resulting in lower mortality of new recruits, from increased adult reproductive output, which increases the number of potential recruits, or from reductions in size and/or age at maturity of the unharvested population, which increases the number of reproductive individuals. I predicted the overcompensatory potential of a population of Carcinus maenas (European green crab) in Bodega Harbor, California, using a series of laboratory and field experiments examining intraspecific pressures of adults on juveniles in the population. This measure of intraspecific pressure was used to predict the overcompensatory potential of the population in response to increased mortality from ongoing removal efforts. This prediction was then assessed using pre- and post-removal surveys of juvenile recruitment in Bodega Harbor compared to nearby populations, testing for evidence of overcompensation. While adult C. maenas in Bodega Harbor had limited negative impacts on juveniles, I concluded it was unlikely to result in overcompensation. Relative juvenile abundance did not statistically increase in removal compared to reference populations, consistent with my conclusion from the experiments. Increases in recruitment rates can occur as a result of efforts to remove non-native species. This increase in recruitment can result in overcompensation, but more commonly results in compensation, where recruitment rates increase relative to pre-removal recruitment but does not result in in the population's abundance exceeding pre-removal levels. However, a detailed and accurate prediction of the response of a population to harvest is time consuming and data intensive. This is not feasible for most efforts to eradicate non-native species, which have the greatest chance of success when enacted rapidly after detection. For my final chapter, I performed a literature review and accompanying statistical analysis to determine if typically available information related to the removal effort (site size, site connectivity, and removal technique) could be used to determine increased risk of compensation for a given effort to remove aquatic invasive species. Compensation was closely linked to unsuccessful removal efforts and was observed only among efforts utilizing physical removal methods. However, the frequency with which compensation occurred varied with the exact technique employed, occurring most frequently in removal utilizing electrofishing. Additionally, evidence of compensation was more frequent among larger removal areas with variable connectivity. While other predictors (temperature, effort, etc) might add to the predicative power, the findings of the review provide criteria for managers to determine the relative risk of compensation prior to the start of removal. Further understanding of how invasive species respond to lethal biotic interactions, including anthropogenically mediated control measures, can aid in assessing the risk of invasion for a given species and inform managers of the risk of complications resulting from removal efforts. While inducible defenses may contribute to the long-term success of an introduced species in their recipient range, my findings did not support the idea that inducible defenses triggered by predator cues contributed to their initial introduction in this case. However, research on other non-native species and offspring of previously naïve prey would allow for a clearer picture of the role of inducible defenses in the invasion process. Compensation resulting from removal efforts does not guarantee failure, and certain characteristics of removal efforts seem to indicate increased risk of compensation. Together these components help identify how biotic interactions surrounding mortality risk of an invading species help shape the trajectory of invasion.

When multiple predators foraging together have different individual consumption rates than predators foraging in isolation, they exhibit non-independent multiple predator effects on prey. I examined multiple predator effects in a system consisting of invasive green crabs (Carcinus maenas L.), native rock crabs (Cancer irroratus Say) and benthic macrofauna prey. First, I examined multiple predator effects when green crabs and rock crabs forage on soft-shell clams (Mya arenaria L.) in different habitat types (sand, sand with artificial seagrass) and assessed the behavioural mechanisms responsible for the observed predation effects. Independent multiple predator effects on prey were detected for most conspecific and heterospecific pairs in both habitat types. In general, crab foraging behaviours were not affected by the presence of another predator. Interactions between predators did not influence foraging behaviours because encounters were infrequent, short in duration and predominantly non-aggressive. A non-independent multiple predator effect on prey (marginally significant) was observed when green crabs foraged with rock crabs in artificial seagrass. This effect, however, could not be explained by the observed crab behaviours. Second, I investigated multiple predator effects when green crabs and rock crabs forage on a soft-sediment macrofauna community. Because crabs did not have significant predation effects on the community throughout the experiment, I did not evaluate multiple predator effects on prey. It is possible that crab predation was not important in regulating the macrofauna community, in which case multiple predator effects were non-existent. Predation may have been suppressed due to a combination of factors, including interactions between predators, harsh environmental conditions or a sub-optimal prey field. Alternatively, my ability to detect significant predation effects may have been hindered because of prey movement in and out of cages or low statistical power. Overall, results from this thesis demonstrate that multiple predator effects on prey may differ with habitat and highlights the importance of conducting behavioural observations to better understand interactions between predators and the resulting consequences for prey. Multiple predator effects on a soft-sediment community should be re-evaluated to assess the importance of these crab species in regulating benthic macrofauna under natural conditions.