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Tethering assays, or the physical restraint of test organisms, has been used in the past to measure selected organisms' response to stressors while removing the observer from the experimental setting. Although informative for monitoring and hypothesis testing, these assays often used microfilaments that have been found to be too invasive or prone to biases given their effects on test organisms' behavior. Here, we describe a new variation of tethering using American oysters (Crassostrea virginica) and illustrate its use in the study of their mortality rates as a result of two stressors: siltation and predation by a non-indigenous species. Our protocol identified a resistant (non-toxic) glue that could be used to attach oysters to stone slabs, thus partially mimicking the natural cementation of the shell to natural substrates. This variation of tethering was harmless and maintained oysters' body position and natural ability to filter feed. Using tethered oysters in separate two-week field cage experiments, we also show how siltation and predation by a non-indigenous species (the European green crab, Carcinus maenas), caused a gradual, easily measurable increase in oyster mortality rates. We argue that this variation of tethering is a cost-effective and advantageous way to monitor or test the effects of these and other stressors on oysters and similar species.

Background Late in the 1990s, the non-indigenous European green crab (Carcinus maenas) colonized the shorelines of eastern Prince Edward Island, in Atlantic Canada. Due to concerns of further spread into productive shellfish habitats, an annual survey was conducted between 2000 and 2013 to detect a potential range expansion of this species. We compiled and analyzed that data and document green crab expansion using records of annual occurrence and relative density. Methods Surveys were conducted during the fall season of each year by deploying baited traps at 29 sites along the island’s two main shorelines (north and south shores). These sites were selected based on areas deemed more likely to be invaded by the green crab. Raw data per site and date was transformed to catch per unit effort (CPUE) to estimate relative abundances. Results Populations of this species showed an uneven westward expansion along the north and south shores. Expansion rates changed among years but, overall, crab abundance was higher and changes in abundance were faster along the south shore than the north shore of the island. The westward expansion continues until this day. Conclusions Based on the information compiled we hypothesize that the dissimilarity in range expansion rate was related to the availability of suitable habitat to sustain large green crab populations along the south shore. We also discuss implications of this expansion for commercial shellfish and native coastal communities.

8 pages, 2 figures, 2 tables, Ocean acidification is expected to affect marine organisms in the near future. Furthermore, abrupt short-term fluctuations in seawater pCO2 characteristic of near-shore coastal regions and high-density aquaculture sites currently have the potential to influence organismal and community functioning by altering animal behaviour. While anti-predator responses in fishes exposed to elevated pCO2 are well documented, such responses in benthic marine invertebrates are poorly studied. We used high frequency, non-invasive biosensors to test whether or not short term (3-week) exposure to elevated pCO2 could impact behavioural responses to the threat of predation in adult Mediterranean mussels from Galicia on the northwestern coast of Spain. Predator alarm cues (crushed conspecifics) resulted in a prolonged (1 h) reduction in the degree of valve opening (−20%) but had no clear effect on overall valve movement activity, while elevated pCO2 did not affect either response. Our results add to the increasing body of evidence suggesting that the effects of end-of-century pCO2 levels on marine animal behaviour are likely weak. Nonetheless, longer-term exposures spanning multiple generations are needed to better understand how ocean acidification might impact behavioural responses to predation in marine bivalves, This project was funded by the Spanish government through the Ministerio de Economía y Competitividad that included European FEDER funds (projects Ref. AGL-2013-45945-R, CTM2016-76146-C3-2-R/CTM2016-76146-C3-1-R)

The growth and spread of non-indigenous green crabs (Carcinus maenas) in Atlantic Canada is of concern to the sustainability of shellfish resources, particularly in areas recently invaded. Commercial green crab fishing has been initiated on Prince Edward Island to help control this species and provide a new resource for inshore fishermen. Developing a soft-shell crab product modelled after the Venetian ‘Moleche’ would provide an economic incentive beyond the existing hard-shell crab bait market. However, answers to questions such as the timing and characteristics of green crab moulting are required. A pilot study conducted in 2014–2015 collected seven groups of crabs and held them in individual compartments for 2–4 weeks to record moulting rates and physical characteristics. We found that a synchronized ‘moulting window’ occurs during July for male crabs. Field experiments in 2015 had an average moulting rate of 34%, with group-specific rates as high as 60%. The same cohort of crabs held in the la...

Atlantic Canada’s American oyster (Crassostrea virginica) beds, while economically and ecologically important, have been in decline over the past few decades. Predation by crabs, in particular by the European green crab (Carcinus maenas), has been proposed as one of the potential causes of such decline. Hence, this study examined oyster mortality levels in multiple beds across Prince Edward Island (PEI) and then experimentally assessed the contribution of green crab predation to oyster mortality. Results from surveys conducted in 10 estuaries across PEI in 2014 indicate that the probability of mortality for small oysters was significantly higher when green crabs were present then in areas without green crabs. This probability of mortality was significantly less when there was the presence of alternative prey like natural mussel beds (Mytilus edulis). The odds of oyster mortality were also higher when beds had rock crabs (Cancer irroratus) compared to beds with no rock crabs. Given the potential importance of green crab predation, its influence was assessed in 2015 using two field experiments with tethered oysters. Our results indicate that odds of small oyster mortality occurring were much higher in green crab inclusion cages than in the open environment and the exclusion cages. These results reaffirm that oysters up to ~40 mm SL are vulnerable to predation, and at least some of the mortality affecting these oysters can be causally attributed to green crab predation. Green crab predation rates upon small oysters are relevant given the economic benefits and ecosystem services provided by these bivalves. They highlight the need for the industry to consider mitigation measures and potentially adapt their oyster growing strategies.