Gastric processing in the Dungeness crab, Cancer magister, during hypoxia

The gastric physiology of the Dungeness crab, Cancer magister, was investigated over a range of oxygen tensions. Postprandial crabs reacted differently to hypoxia compared with unfed animals. The bradycardic response in postprandial animals was reduced, suggesting a summation of responses with feeding. A similar pattern was observed for ventilation rate. In unfed animals ventilation rate increased slightly as oxygen levels declined, but dropped significantly in oxygen tensions below 3.2 kPa, whereas in postprandial crabs it increased significantly in the lower oxygen regimes. Gastric processing of the meal was followed using a fluoroscope. Pyloric contraction rates were maintained during mild hypoxia, but decreased in 5.3 kPa oxygen tension and below. This led to an increase in clearance times of digesta from the foregut, midgut and hindgut regions. The slowing of gastric processing in the lower oxygen tensions suggested that the animals were unable to maintain their internal oxygen concentration. A significant reduction in efficiency of assimilation only occurred in the lowest oxygen regime tested (1.6 kPa). The range of hypoxia where gastric processing was affected corresponded closely to the levels of oxygen that modulate the foraging behaviour of C. magister. By using both physiological and behavioural mechanisms C. magister can maintain digestive processes, even in severely oxygen depleted environments.     

Gastrin/cholecystokinin-like immunoreactive peptides in the Dungeness crab, Cancer magister (Dana): immunochemical and biological characterization

The purpose of this investigation was to characterize a gastrin/cholecystokinin-like immunoreactant (G/CCK-LI) extractable from the crab, Cancer magister. G/CCK-LI was extracted best in boiling water and was found mainly in the stomach, hemolymph and carapace. A relatively large immunoreactive peptide in the stomach and apparently smaller forms in the hemolymph and carapace were separated by Sephadex G-50 fractionation. Anion-exchange chromatography further fractionated the stomach form into three major peaks. The crab material cross-reacted with three antisera specific for the common C-terminus of gastrin/CCK, but cross-reacted much less with three antisera directed against other portions of the gastrin molecule. Partially purified crab stomach G/CCK-LI inhibited the binding of labeled CCK to mouse brain G/CCK receptors but not to rat pancreatic CCK receptors. The crab peptide did not stimulate rat gastric acid or rat pancreatic amylase secretion. These results indicate that the crab peptides are structurally similar to, but distinguishable from, the bioactive C-terminal amino acid sequence common to gastrins and CCKs.     

Influence of hypoxia on bacteremia in the Dungeness crab, Cancer magister

We examined the possibility that decreased environmental oxygen can elevate the levels of indigenous bacteria in the hemolymph of Cancer magister. Crabs were exposed to air-saturated and hypoxic (50% air-saturation) water for 3 days and levels of culturable bacteria in hemolymph were measured every 24 h as the total number of colony-forming units (CFU) per milliliter of hemolymph. Bacteremia increased after 24 h of exposure to hypoxia and persisted for 72 h, whereas crabs exposed to normoxia had no measurable change in number of culturable bacteria. The predominant persistent bacteria in the hemolymph was isolated and identified by DNA sequence-based methods as Psychrobacter cibarus. Crabs were injected with P. cibarus or with buffered saline as a control after 3 h of hypoxia. Levels of culturable bacteria were significantly higher in hypoxic crabs than in normoxic ones (about 2500 versus 1000 CFU ml(-1) 80 min post-injection, respectively), and circulating levels of oxygen were significantly reduced in infected animals compared to uninfected ones after 48 h in hypoxia and after 72 h in air-saturated water post-injection. These data demonstrate that P. cibarius is present in Dungeness crabs, that environmental hypoxia can dramatically elevate levels of persistent bacteria, and that hypoxia in the presence of hemolymph bacteria may ultimately reduce immune and respiratory ability.     

Peptidergic modulation of cardiovascular dynamics in the Dungeness crab,Cancer magister

Decapod crustacean pericardial organs contain extensive neurohormonal reserves which can be released directly into the haemolymph to act as physiological modulators. The present paper concerns the in vivo effects of two pericardial peptides, proctolin and crustacean cardioactive peptide, on cardiovascular dynamics in the crab Cancer magister. Infusion of proctolin into the pericardial sinus caused a slight decrease in heart rate concurrent with a large increase in cardiac stroke volume. It decreased haemolymph flow anteriorly through the paired anterolateral arteries and increased flow posteriorly and ventrally through the posterior aorta and sternal artery, respectively. The threshold for responses occurred at circulating concentrations of 10^-9 mol·l^-1, and haemolymph flows remained elevated for up to 30 min after peptide infusion. The effects of crustacean cardioactive peptide were less dramatic. Heart rate was not affected but a significant increase in stroke volume was observed. Crustacean cardioactive peptide increased haemolymph flow through the anterolateral arteries and increased scaphognathite rate. The threshold for crustacean cardioactive peptide activity was higher than for proctolin (10^-7 mol·l^-1 and 10^-6 mol·l^-1) but the responses to crustacean cardioactive peptide were of longer duration. The effects of proctolin on regional haemeolymph distribution in Cancer magister closely resemble the cardiovascular responses of this species when exposed to hypoxic conditions. These peptides may be implicated as cardiovascular regulators during environmental perturbations.     

Responses of bender apodeme tension receptors in the Dungeness crab,Cancer magister

Receptors monitoring tension are located on the apodemes of the bender (productor propoditis) muscle of walking legs and chelipeds of the Dungeness crab, Cancer magister. The bipolar neurons form a nerve, the bender apodeme sensory nerve (BASN), which joins the CP1 chordotonal organ nerve proximally, BASN units exhibit spontaneous activity at rest, and fire bursts of action potentials to rapid passive reduction of the carpopodite-propodite joint. Isometric contraction of the bender muscle results in BASN output that is directly related to force. Afferent activity ceases upon quick release of isometric tension, when tension drops to zero, and the unloaded muscle contracts isotonically.     

A ciliate infection (Paranophrys sp.) in laboratory-held Dungeness crabs,Cancer magister

A ciliate infection ascribed to the genus Paranophrys was found in early adult Dungeness crabs, Cancer magister, held in laboratory bioassay facilities, and in outdoor flow-through tanks in Newport, Oregon. Infections proved lethal as ciliates multiplied to high densities in the blood and tissues causing degeneration and necrosis of musculature. All infected crabs had recent wounds through the exoskeleton which probably provided a portal of entry, and the elapsed time between infliction of a wound and gross symptoms of stress was estimated at 9 days although animals died as late as 26 days. The nature of Paranophrys infections in C. magister, i.e., as an opportunistic pathogen of cultured animals and/or a disease in wild populations of this crab is discussed.     

The competitive and predatory impacts of the nonindigenous crab Carcinus maenas (L.) on early benthic phase Dungeness crab Cancer magister Dana

We evaluate the potential competitive and predatory impacts of nonindigenous European green crab Carcinus maenas on native Dungeness crab Cancer magister in the northeast Pacific. The coastal estuaries of Washington State, USA, provide appropriate habitat for recently introduced green crab, yet these areas are important nursery grounds for Dungeness crab and contribute greatly to the coastal crab fishery. Juvenile Dungeness crabs are dependent on limited intertidal epibenthic shell for refuge habitat during early benthic life and experience increased mortality on open sand and mud as a result of predation by fish and birds. Early juveniles throughout the subtidal are similarly at risk due to predation by fish and especially adult conspecifics. Laboratory experiments and infrared video observations revealed that juvenile green crab displace Dungeness crab of equal size from shelters during one-on-one competition. Green crab also consistently win nocturnal foraging trials in which the species compete for fresh, damaged clams. Field and laboratory enclosure experiments show that juvenile Dungeness crab emigrate from oyster shell habitat as a result of competition and predation by adult green crab. Depending on the extent to which the two species overlap, interactions with the dominant nonindigenous species could have a negative influence on juvenile Dungeness crab survival and could conceivably impact recruitment to the fishery. However, current evidence indicates that the distribution of green crab in Washington State is far removed from nursery areas of Dungeness crab.     

A systemic microbial disease in the Dungeness crab,Cancer magister,caused by a Chlamydia-like organism

A Chlamydia-like agent was diagnosed in Dungeness crabs, Cancer magister, collected from Willapa Bay and northern Puget Sound, Washington. Over a 5-year period, infected crabs were found during the months of December through March. Infection by the organism is systemic, resulting in greatly hypertrophied cells with appressed nuclei. Unlike Chlamydia or other Chlamydia-like organisms, the present microbe has a great affinity for connective tissue or connective tissue cells rather than epithelial cells. Transmission electron microscopy revealed the presence of several developmental stages that are similar to chlamydial stages, in addition to what appear to be slightly altered or aberrant stages.     

Isolation and characterization of polymorphic microsatellite loci from the Dungeness crab Cancer magister

The Dungeness crab, Cancer magister, is the focus of one of the most intensely harvested fisheries in North America. Given its economic importance, there is considerable interest in assessing the degree and spatial pattern of genetic structure in C. magister. To that end, we developed a series of 17 hypervariable microsatellite loci. Six of these 17 loci could be amplified in a single multiplex PCR reaction. Using dye‐labelled primers all six loci can be coamplified and scored simultaneously on an automated sequencer. The ability to multiplex multiple loci greatly increases the ease and speed of genotyping for this species.     

A mycosis caused by Lagenidium sp. in laboratory-reared larvae of the Dungeness crab,Cancer magister,and possible chemical treatments

A mycosis in larvae of the Dungeness crab, Cancer magister, was observed about 48 hr after they had molted to second-stage zoeae in the laboratory during a chronic bioassay. The fungus was of the genus Lagenidium and caused about 40% mortality of larvae before the test was terminated. Hyphal development and subsequent sporulation of the fungus was observed over a range of 0–32‰ salinity, with an optimum around 16‰. Bioassays of eight fungicides and the herbicide trifluralin, using the larvae and fungus in separate tests, were performed to explore means of chemical treatment. Trifluralin was the most satisfactory of the chemicals screened, with captan, Cu²⁺, and benomyl also giving good results for certain treatment intervals of up to 96 hr. Malachite green was toxic to larvae at concentrations inhibiting the fungus.     

Evidence for interannual variation in genetic structure of Dungeness crab (Cancer magister) along the California Current System

Using a combination of population‐ and individual‐based analytical approaches, we provide a comprehensive examination of genetic connectivity of Dungeness crab (Cancer magister) along ~1,200 km of the California Current System (CCS). We sampled individuals at 33 sites in 2012 to establish a baseline of genetic diversity and hierarchal population genetic structure and then assessed interannual variability in our estimates by sampling again in 2014. Genetic diversity showed little variation among sites or across years. In 2012, we observed weak genetic differentiation among sites (F_ST range = ?0.005–0.014) following a pattern of isolation by distance (IBD) and significantly high relatedness among individuals within nine sampling sites. In 2014, pairwise F_ST estimates were lower (F_ST range = ?0.014–0.007), there was no spatial autocorrelation, and fewer sites had significant evidence of relatedness. Based on these findings, we propose that interannual variation in the physical oceanographic conditions of the CCS influences larval recruitment and thus gene flow, contributing to interannual variation in population genetic structure. Estimates of effective population size (N_e) were large in both 2012 and 2014. Together, our results suggest that Dungeness crab in the CCS may constitute a single evolutionary population, although geographically limited dispersal results in an ephemeral signal of IBD. Furthermore, our findings demonstrate that populations of marine organisms may be susceptible to temporal changes in population genetic structure over short time periods; thus, interannual variability in population genetic measures should be considered.     

Respiratory and digestive responses of postprandial Dungeness crabs, Cancer magister, and blue crabs, Callinectes sapidus, during hyposaline exposure

Respiratory responses and gastric processing were examined during hyposaline exposure in two crab species of differing osmoregulatory ability. The efficient osmoregulator, Callinectes sapidus, displayed an immediate increase in oxygen uptake when exposed to low salinity in isolation. In contrast, the weak osmoregulator, Cancer magister, showed no change in oxygen uptake upon acute exposure (<6 h), but slight increases in oxygen uptake tended to occur over longer time scales (12–24 h). These changes were likely attributable to an increase in avoidance activity after 6 h hyposaline exposure. Following feeding in 100% SW, oxygen uptake doubled for both species and remained elevated for 15 h. When postprandial crabs were exposed to low salinities, C. sapidus were able to sum the demands of osmoregulation and digestion. Thus, gastric processes continued unabated in low salinity. Conversely, postprandial C. magister prioritized responses to low salinity over those of digestion, resulting in a decrease in oxygen uptake when exposed to low salinity. This decrease in oxygen uptake corresponded to a reduction in the rate of contraction of the pyloric stomach and a subsequent doubling of gastric evacuation time. The current study is one of the few to illustrate how summation or prioritization of competing physiological systems is manifested in digestive processes.     

Larval settlement of the nemertean egg predator Carcinonemertes errans on the Dungeness crab,Metacarcinus magister

Due to their habitat specificity, marine parasites present excellent systems for studying the processes and patterns of larval settlement. Settlement of Carcinonermertes errans, an egg predator of the Dungeness crab, is described here for the first time. Upon contact with a host individual, competent larvae of C. errans settled on the crab's exoskeleton and migrated under the abdominal flap within 24 h. When removed from the host, recently settled worms retained their larval characteristics. After 48 h on the host, however, metamorphosis proceeded and larvae became juvenile worms. Additional field studies showed that competent larvae were present in the waters of the Coos Bay Estuary during the months of August through early November, could infect crab hosts directly from the water column, and exhibited density‐dependent gregarious settlement.     

Larval development of Euphausia superba Dana, 1852 and a phylogenetic analysis of the Euphausiacea

Literature data and new investigations by SEM of selected ontogenetic stages of the Antarctic Krill, Euphausia superba Dana, 1852 revealed morphological characters that are either missing in, or significantly changed towards, the adult. Besides adult features, such ontogenetic characters enabled us to propose a hypothesis of the phylogenetic relationships of and within the Euphausiacea on the basis of a computer aided cladistic analysis. These are of the form (Bentheuphausia amblyops + Euphausiidae = ('Thysanopoda' + Nematobrachion + Euphausiinae = (Meganyctiphanes norvegica + Euphausiini, new name + Nematoscelini, new name = (Nyctiphanes + Nematoscelina, new name)))). From this analysis the taxon names 'Euphausiina', 'Nematoscelini', and 'Nematoscelina' are introduced for in-groups of the taxon Euphausiacea as representing monophyletic units. The position of a set of ontogenetic characters remains relatively uncertain due to the still unknown larval development of Bentheuphausia amblyops (G. O. Sars, 1883).