Epidemiology of bitter crab disease (Hematodinium sp.) in snow crabs Chionoecetes opilio from Newfoundland, Canada

The parasitic dinoflagellate Hematodinium sp. causes a condition known as bitter crab disease (BCD) in snow crabs Chionoecetes opilio and Tanner crabs C. bairdi. As the name of the condition implies, crabs infected with BCD are unmarketable due to their bitter flavor. We surveyed the distribution of BCD in 3 regions within the snow crab fishery of Newfoundland from 1997 to 2003. Over time, the disease has become firmly established in Conception and Bonavista Bays and persists at low levels on the Avalon fishing grounds. An epizootic occurred within Bonavista and Conception Bays in 1999 and persisted in Conception Bay in 2000, reaching prevalences of over 2% to 9% in trapped and trawled male crabs and from 19 to 26% in trawled and trapped female crabs, respectively. Infections were highest in females and small males, i.e. the unfished and pre-recruit portions of the fishery. In a mortality study, all of the naturally infected crabs died and 50% of the experimentally inoculated crabs died. Patterns in the molting cycle and prevalence of infection indicate that transmission occurs during the post-molt condition, and that overt infections probably develop 2 to 4 mo later with mortalities occurring at least 3 to 4 mo thereafter. The hydrography of this bay may have contributed to the epizootic as infections were centered within the deeper confines of the bay. Analysis of various abiotic factors uncovered a significant positive association between prevalence, depth and mud/sand substrates; the nature of this relationship was not apparent but may be related to diet or alternate hosts. Lastly, given the increase in BCD in snow crabs in Newfoundland, we recommend that fishery management programs for Chionoecetes fisheries employ non-selective gear to monitor for Hematodinium infections in female and juvenile crabs because these under-sampled members of the population may forewarn of impending recruitment declines that might otherwise remain unexplained.     

Monitoring the prevalence of the parasitic dinoflagellate Hematodinium sp. in snow crabs Chionoecetes opilio from Conception Bay,Newfoundland

Bitter crab disease (BCD) of snow crabs Chionoecetes opilio is caused by a parasitic dinoflagellate, Hematodinium sp. In Newfoundland's commercial fishery, infected snow crabs are identified using visual, macroscopic signs of disease for separation prior to processing. We estimated the sensitivity and specificity of gross, macroscopic diagnosis of Hematodinium sp. by comparing these results with microscopic examination of prepared hemolymph smears. The sensitivity of a diagnostic test is the probability that the test will yield a positive result given that the animal has the disease. The specificity is the probability of a negative result given the animal is not diseased. In October 1998, we conducted a design-based survey using cluster sampling in 2 strata. Over 10 000 snow crabs from pot and trawl surveys were examined macroscopically for BCD. In addition, over 350 crabs were randomly examined microscopically for disease. The double sampling resulted in an estimated sensitivity of 52.7% and an estimated specificity of 100%. That is, a positive result from macroscopic examination is definitive, if the observer is well trained, but macroscopic examination will fail to detect infections in crabs with borderline clinical signs of disease. The prevalence estimated from macroscopic observations (p(st) = 2.24%) was corrected for misclassification by dividing p(st) by the estimated sensitivity (0.527), giving a corrected estimate of 4.25%. The use of double sampling provides for efficient estimation of prevalence in that large numbers of crabs can be quickly examined for gross signs of infection and the results corrected for misclassification based on a limited number of observations with a better, but time-consuming test. In addition, the prevalence of macroscopically infected male crabs was lower in a trap survey (0.57%) compared to a trawl survey (1.59%). In the trawl survey, female crabs had a significantly higher prevalence of macroscopically diagnosed infections than males (6.34%). The prevalence of BCD has shown an alarming increase since it was first detected in Newfoundland during the early 1990s. Transmission and mortality studies are warranted to better understand the effect of the disease on its commercially important host.     

New microsatellite markers for the snow crab Chionoecetes opilio (Brachyura: Majidae)

Nine novel microsatellite loci were isolated from Chionoecetes opilio by screening an enriched genomic library using nonradioactive polymerase chain reaction techniques, and the polymorphisms were examined to estimate genetic variability. The genetic variabilities varied depending on the locus. All loci were found to be polymorphic with an average of 9.7 alleles per locus (range 3–25). The observed and expected heterozygosities ranged from 0.80 to 0.98 and from 0.56 to 0.95, respectively. Five loci showed significant Hardy–Weinberg disequilibrium at the P < 0.05 level. The high variabilities revealed in this study suggest that these microsatellite loci should provide useful markers for genetic variation monitoring of C. opilio.     

Prevalence of bacteria in the spermathecae of female snow crab, Chionoecetes opilio (Brachyura: Majidae)

The spermathecal contents of primiparous, multiparous and barren female snow crabs were observed by a scanning electron microscope. Bacteria colonies were observed with a significantly higher frequency in the spermathecae of primiparous and old barren females. Bacteria infect the spermathecae and destroy the spermatophores and spermatozoa inside. These observations suggest that bacteria in the spermathecae do not exclude opportunistic microbes by modifying pH of the medium as suggested in the literature. The prevalence of bacteria in primiparous and old barren females suggests that they infect individuals with a weak anti-microbial protection. The absence of bacteria in the highly acidic seminal fluid derived from males upon copulation suggests that it may provide anti-microbial protection. Bacteria do not seem to be able to survive in an anaerobic environment for a long period.     

A review of factors contributing to the decline of Newfoundland and Labrador snow crab (Chionoecetes opilio)

The Newfoundland and Labrador (NL) snow crab resource, presently the basis of the most important commercial fishery in the region, is in decline. Short-, mid-, and long-term recruitment prospects are deemed poor in most areas. Fishery declines have been most apparent in the north, beginning in the mid- to late 2000s, but are expected to begin in the more productive southern areas in the near future. A multitude of emergent theories to explain the resource decline have been hypothesized as contributing factors, including fishing, trawling impacts, seismic activities, disease, predation, and increasing temperature. This study comprehensively reviews and qualitatively relates the results of recent research and literature on each of these factors. We find that several factors may be contributing to a lack of recruitment in the stock, but diminishing productivity resulting from a warming oceanographic regime is the primary cause of the resource decline. Further, we postulate that trends occurring in the snow crab stock are indicative of a broader-scale ecological regime shift occurring along the NL shelf.     

Evidence for a permanent establishment of the snow crab (Chionoecetes opilio) in the Barents Sea

In the Atlantic the snow crab (Chionoecetes opilio) is naturally distributed on the northwestern side, i.e. eastern Canada and west Greenland. Until recently, there have been no observations of snow crab in eastern Atlantic. However, in 1990s single and occasional reports were made of crabs captured in the eastern part of the Barents Sea, presumably introduced through ballast water. Special attention during the annual bottom-trawl surveys in the Barents Sea during February 2004–2006 were given to include recordings of snow crab to evaluate if the introduced species has succeeded to establish a self-sustaining population in this region. Recordings of snow crabs were systematically noted and biological measurements carried out. The results confirm previous Russian observations of snow crabs in the northern region of Gåsebanken. In addition, a significant number of crabs were also found in the central region of the Barents Sea, mainly in deeper waters from 180 to 350 m depth. The sizes ranged from 14 to 136 mm carapace width. All females above 70 mm were berried with fertilised eggs. A major fraction (31% in 2005; 76% in 2006) of the crabs consisted of juveniles below 50 mm CW, providing evidence for successful recruitment. The small-sized crabs were exclusively found in Gåsebanken, identifying the main recruiting area at present for snow crab in the Barents Sea. The results obtained show that the snow crab is now adapted to the northeast Atlantic.     

Moulting and growth in earlier and later moulters of adolescent male snow crabs (Chionoecetes opilio) (Brachyura: Majoidea) under laboratory conditions

Growth and moulting of male adolescent snow crabs were investigated in a laboratory culture experiment. The moulting season of adolescents was separated into two groups of earlier and later, and they were regarded as normal and skip moulters, respectively. The existence of male snow crab skip moulters was first demonstrated in the Sea of Japan. Although the precise moulting season of the earlier moulter could not be revealed owing to the captive method, the later moulter is suspected to moult approximately 1 year after the earlier moulter. Earlier and later moulters were not different in growth of carapace width or chela height during the pubertal and terminal moult. Our moult and growth results of earlier and later moulters will be useful for predicting recruitment to legally fishable (>90 mm carapace width) or valuable (hard-shelled adult) populations of male snow crab in the Sea of Japan.     

Biochemical indicators of muscle growth in the snow crab Chionoecetes opilio (O. Fabricius)

This study examined the relationships between muscle growth rate, the activity of metabolic enzymes and the RNA:DNA ratio, in adult snow crabs Chionoecetes opilio. After moulting, crabs were assigned to three feeding rations to attain a range of tissue growth rates. Muscle growth rate, estimated by the variation in dry tissue content per ml of merus of the first walking leg, was positively correlated with changes in muscle cell number, as evaluated by the DNA content per ml of merus. However, no significant correlation was detected between growth rate and the variation in muscle cell size, the latter being estimated by the change in the protein:DNA ratio. This is due to the fact that, in starved crabs, a reduction in the number of cells is partly compensated by a size increment of the remaining ones. This phenomenon also weakened the overall relationship between muscle growth rate and the phosphofructokinase (PFK) capacity per ml of merus. The simple correlation between those two variables was significantly positive for animals which increased their mass of muscle but insignificant for those which were loosing muscle mass. The lactate dehydrogenase (LDH), citrate synthase (CS) and cytochrome c oxidase (CCO) capacity per ml of merus did not match growth rate. The significant simple correlations that were detected between growth rate and the various enzyme activity expressed per g of protein, per μg of DNA and per g of dry mass did not hold when partial correlations were computed. Variations in muscle cell size were related to adjustments in the quantity of RNA per cell, as depicted by the RNA:DNA ratio. Since muscle growth was not correlated with the variation in muscle cell size, it was not correlated with the RNA:DNA ratio either.     

Isolation and characterization of polymorphic microsatellite markers in the red snow crab (Chionoecetes japonicus)

A total of 12 polymorphic microsatellite DNA loci were isolated from the red snow crab, Chionoecetes japonicus (Brachyura: Majidae), one of important fisheries resources in the Far East. The number of alleles observed at each locus ranged from two to 19, with the observed and expected heterozygosities of 0.125-0.875 and 0.156-0.949, respectively, suggesting these loci to be a useful molecular marker for population analysis in this species. Of the 12 loci, seven also were available for genotyping of the snow crab, Chionoecetes opilio, implying these loci as a useful molecular marker in the genus Chionoecetes.     

First record of the decapod-egg predator Ovicides paralithodis (Nemertea,Carcinonemertidae) from the snow crab Chionoecetes opilio (Decapoda,Brachyura)

The carcinonemertid monostiliferan Ovicides paralithodis Kajihara and Kuris, 2013 was originally described as an egg predator of the red-king crab Paralithodes camtschaticus (Tilesius, 1815) in the Sea of Okhotsk, the Bering Sea, and the Gulf of Alaska. In the present study, several carcinonemertid specimens were obtained from the egg mass of the snow crab Chionoecetes opilio (O. Fabricius, 1788) in the Sea of Japan. Partial sequences of the mitochondrial cytochrome c oxidase subunit I gene (COI) determined from two specimens of the carcinonemertid were identical with a barcode sequence from the holotype of O. paralithodis, indicating that the host range of the species covers at least the two decapod species, P. camtschaticus and C. opilio.     

Variable mate-guarding time and sperm allocation by male snow crabs (Chionoecetes opilio) in response to sexual competition, and their impact on the mating success of females.

Two laboratory experiments investigated mate guarding and sperm allocation patterns of adult males with virgin females of the snow crab, Chionoecetes opilio, in relation to sex ratio. Although females outnumbered males in treatments, operational sex ratios were male-biased because females mature asynchronously and have a limited period of sexual attractiveness after their maturity molt. Males guarded females significantly longer as the sex ratio increased: the mean time per female was 2.9 d in a 2 males:20 females treatment compared to 5.6 d in a 6 males:20 females treatment. Female injury and mortality scaled positively to sex ratio. Males that guarded for the greatest number of days were significantly larger, and at experiment's end had significantly smaller vasa deferentia, suggesting greater sperm expense, than males that guarded for fewer days. In both experiments, the spermathecal load (SL)--that is, the quantity of ejaculate stored in a female's spermatheca--was independent of molt date, except in the most female-biased treatment, where it was negatively related. The SL increased as the sex ratio increased, mainly because females accumulated more ejaculates. However, similarly sized males had smaller vasa deferentia and passed smaller ejaculates, such that, at a given sex ratio, the mean SL was 55% less in one experiment than in the other. Some females extruded clutches with few or no fertilized eggs, and their median SL (3-4 mg) was one order of magnitude smaller than that of females with well-fertilized clutches (31-50 mg), indicating sperm limitation. Males economized sperm: all females irrespective of sex ratio were inseminated, but to a varying extent submaximally; each ejaculate represented less than 2.5% of male sperm reserves; and no male was fully exhausted of sperm. Sperm economy is predicted by sperm competition theory for species like snow crab in which polyandry exists, mechanisms of last-male sperm precedence are effective, and the probability that one male fertilizes a female's lifetime production of eggs is small.     

Bitter crab syndrome in two species of king crabs from the Sea of Okhotsk

The disease caused by parasitic dinoflagellates of the genus Hematodinium was found in the red Paralithodes camtschaticus and blue P. platypus king crabs from the Sea of Okhotsk. The hemolymph of diseased crabs was cream colored, opaque, and dense. Numerous dinokaryotic trophonts and multinucleate plasmodia of the parasite were revealed in the hemolymph and internals of diseased animals. The parasitic infection was recorded in females and juvenile males from August to mid-October.     

Evidence of Antibacterial Activities in Peptide Fractions Originating from Snow Crab (Chionoecetes opilio) By-Products

Antibacterial peptide fractions generated via proteolytic processing of snow crab by-products exhibited activity against Gram-negative and Gram-positive bacteria. Among the bacterial strains tested, peptide fractions demonstrated inhibitory activity against the Gram-negative bacteria such as Aeromonas caviae, Aeromonas hydrophila, Campylobacter jejuni, Listonella anguillarum, Morganella morganii, Shewanella putrefasciens, Vibrio parahaemolyticus and Vibrio vulnificus and against a few Gram-positive bacteria such as Listeria monocytogenes, Staphylococcus epidermidis and Streptococcus agalactiae. The principal bioactive peptide fraction was comprised mainly of proteins and minerals (74.3 and 15.5%, respectively). Lipids were not detected. The amino acid content revealed that arginine (4.6%), glutamic acid (5.3%) and tyrosine (4.8%) residues were represented in the highest composition in the antibacterial peptide fraction. The optimal inhibitory activity was observed at alkaline pH. The V. vulnificus strain, most sensitive to the peptide fraction, was used to develop purification methods. The most promising chromatography resins selected for purification, in order to isolate peptides of interest and to carry out their detailed biochemical characterization, were the SP-Sepharose? Fast Flow cation exchanger and the Phenyl Sepharose? High Performance hydrophobic interaction media. The partially purified antibacterial peptide fraction was analyzed for minimum inhibitory concentration (MIC) determination, and the value obtained was 25 μg ml^?1. Following mass spectrometry analysis, the active peptide fraction seems to be a complex of molecules comprised of several amino acids and other organic compounds. In addition, copper was the main metal found in the active peptide fraction. Results indicate the production of antibacterial molecules from crustacean by-products that support further applications for high-value bioproducts in several areas such as food and health.     

Metamorphosis season from megalopa to the first crab stage in snow crab Chionoecetes opilio and red snow crab C. japonicus (Crustacea,Decapoda, Majoidea) in the Sea of Japan,estimated from captive culture

ABSTRACT

Metamorphosis season of megalopae to the first crab stage in snow crab Chionoecetes opilio and red snow crab C. japonicus was inferred by culturing wild-born megalopae collected from the Sea of Japan. Metamorphosis occurred from late June to late July (mainly in July) in snow crab, and from early July to early October (mainly from August to September) in red snow crab. The number of days required from the time of collection to metamorphosis was less than the intermoult period previously reported for snow crab megalopae. However, the developmental period of the megalopae was estimated as substantially longer in red snow crab than in snow crab. Previous studies have shown that the hatching season and the period of the zoeal stage in both two species are similar. These results suggest that a different metamorphosis season between the two species would be due to a difference in their megalopal intermoult period.     

Pattern and causes of a temperature-dependent gradient of size at terminal moult in snow crab (Chionoecetes opilio) along West Greenland

Geographic variation in size of male and female snow crabs (Chionoecetes opilio) was investigated along the west coast of Greenland to test the hypothesis that size at terminal molt (=adulthood) is temperature dependent. A total of 81,490 snow crabs were collected in small-mesh traps in Disko Bay (68–69°N) and six sites near Sisimiut (66–67°N) in May and June from 2000 to 2005. Average bottom temperature over the study period ranged from −0.8 to 3.2°C across the sampling sites. Mean carapace width was positively correlated with temperature in both sexes, a pattern often described as a converse Jame’s cline. We infer that temperature per se is the causative factor and discount season length, food availability or density as ultimate causes of the cline. Temperature effects on body size of crabs apparently result from a change in the number of instars before terminal molt. This interpretation is supported by size frequency analysis showing that in general crabs were larger at instar in a colder than in a warmer site. We briefly discuss the implications of our findings for population reproductive potential and the effectiveness of a fixed legal size limit in protecting some adult males from exploitation at different temperature regimes.     

The moulting cycle and changes in body density in larvae of the snow crab Chionoecetes opilio (Brachyura: Majoidea) under laboratory conditions

The moulting cycle and the time course of changes in body density from hatching to the end of the megalopal stage in snow crab (Chionoecetes opilio) larvae were investigated in laboratory-reared specimens. Morphological changes in the epidermis and cuticle were photographically documented to characterize the moult-cycle stages: A–B (postmoult), C (intermoult), D (premoult) and E (ecdysis). Moult-stage characteristics were based on a microscopical examination of integumental modifications, particularly of the telson. During stages A–C, the larval cuticle changed from a spongy structure to become conspicuously thicker and more solid in appearance. In stage D, the epidermis retracted from the cuticle and new setae and appendages were formed. The body densities of larval snow crabs were lowest just after moulting; they increased greatly during stage C, and then gradually increased to reach a plateau at 1.0897–1.0931 g cm^?3 during stage D. Over the whole larval period, they have a density greater than that of seawater. These observations will assist in understanding of larval distribution and transport in snow crabs in their natural habitat, and provide a useful tool to determine the developmental stages of larvae sampled from the plankton and from larval cultures.     

First observations of temperature tolerances of adult male snow crab (Chionoecetes opilio) from the Barents Sea population and the effects on the fisheries strategy

This study investigates the effects of temperature on the survival, food intake, oxygen consumption and growth during long-term live holding of captive male snow crab (Chionoecetes opilio) (average?=?0.7?kg). The crabs were held at three different temperatures, 3, 6 and 9°C. The trials were done using groups of snow crabs held in nine land-based holding tanks (three replicates per temperature treatment). The results showed that temperature had a significant effect on survival. The survival rate at 3°C (61%) was significantly higher than at 6°C (33%) and at 9°C (28%). Specific oxygen consumption rates of unfed crab at 6°C were significantly higher than at 9°C and 3°C. In summary, the current study shows that the Barents Sea snow crab have a narrow temperature range in which they thrive compared with the Barents Sea red king crab (Paralithodes camtschaticus). Barents Sea snow crab has similar metabolic and physiological attributes to other major snow crab populations. In conditions when ambient temperatures are approximately 6°C, it may prove beneficial for animal welfare and also be financially advantageous to reduce ambient water temperatures in live snow crab holding facilities on boat or on land.     

Estimating the maximum sustainable yield of snow crab (Chionoecetes opilio) off Tohoku,Japan via a state-space stock assessment model with time-varying natural mortality

A maximum sustainable yield (MSY) obtained by maintaining or restoring fish stock levels is a tangible benefit of ecosystem services. Snow crab (Chionoecetes opilio) off Tohoku, Japan has been managed by a total allowable catch since 1996, although the abundance has not increased. Surprisingly, there was no increase after 2011, when fishing pressure was greatly reduced because of the Great East Japan earthquake. This implies that some of the crab's biological characteristics, such as recruits, natural mortality coefficient (M) and terminal molting probabilities (p), might have changed. We developed “just another state-space stock assessment model” to estimate the MSY of the snow crab off Tohoku considering interannual variations in M and p. The multimodel inference revealed that M increased from 0.2 in 1997 to 0.59 in 2018, although it did not vary according to instars, sex or terminal molt. The parameter p also increased by 1.34–2.46 times depending on the instar growth stages from 1997 to 2018. We estimated the MSYs in three scenarios, which changed drastically if M and p were set as they were in the past or at the current values estimated from this study. This result indicated that the MSY of snow crab would also vary with time based on their time-varying biological characteristics.     

Predicting the distribution and ecological niche of unexploited snow crab (Chionoecetes opilio) populations in Alaskan waters: a first open-access ensemble model

Populations of the snow crab (Chionoecetes opilio) are widely distributed on high-latitude continental shelves of the North Pacific and North Atlantic, and represent a valuable resource in both the United States and Canada. In US waters, snow crabs are found throughout the Arctic and sub-Arctic seas surrounding Alaska, north of the Aleutian Islands, yet commercial harvest currently focuses on the more southerly population in the Bering Sea. Population dynamics are well-monitored in exploited areas, but few data exist for populations further north where climate trends in the Arctic appear to be affecting species' distributions and community structure on multiple trophic levels. Moreover, increased shipping traffic, as well as fisheries and petroleum resource development, may add additional pressures in northern portions of the range as seasonal ice cover continues to decline. In the face of these pressures, we examined the ecological niche and population distribution of snow crabs in Alaskan waters using a GIS-based spatial modeling approach. We present the first quantitative open-access model predictions of snow-crab distribution, abundance, and biomass in the Chukchi and Beaufort Seas. Multi-variate analysis of environmental drivers of species' distribution and community structure commonly rely on multiple linear regression methods. The spatial modeling approach employed here improves upon linear regression methods in allowing for exploration of nonlinear relationships and interactions between variables. Three machine-learning algorithms were used to evaluate relationships between snow-crab distribution and environmental parameters, including TreeNet, Random Forests, and MARS. An ensemble model was then generated by combining output from these three models to generate consensus predictions for presence-absence, abundance, and biomass of snow crabs. Each algorithm identified a suite of variables most important in predicting snow-crab distribution, including nutrient and chlorophyll-a concentrations in overlying waters, temperature, salinity, and annual sea-ice cover; this information may be used to develop and test hypotheses regarding the ecology of this species. This is the first such quantitative model for snow crabs, and all GIS-data layers compiled for this project are freely available from the authors, upon request, for public use and improvement.     

Biomolecular characterization of allergenic proteins in snow crab (Chionoecetes opilio) and de novo sequencing of the second allergen arginine kinase using tandem mass spectrometry

Snow crab (Chionoecetes opilio) proteins have been recognized as an important source of both food and occupational allergens. While snow crab causes a significant occupational allergy, only one novel allergen has recently been fully characterized. The muscle proteins from snow crab legs were profiled by SDS-PAGE. Several of these proteins were characterized using tandem mass spectrometry. Five proteins were identified; sarcoplasmic Ca-binding (20kDa), arginine kinase (40), troponin (23kDa) and α-actine (42kDa) and smooth endoplasmic reticulum Ca(2+)ATPase (113kDa). Immunoblotting using serum of sixteen allergic patients resulted in strong reactivity with the 40-kDa protein in seven patients (43%). This protein was purified by chromatography and subsequently de novo sequenced using matrix assisted laser desorption ionization and electrospray tandem mass spectrometry. We identified a second important allergen, arginine kinase, in snow crab, designated Chi o 3. Based on identity and homology analysis, using bioinformatics tools, a signature peptide was identified as a chemical surrogate for arginine kinase. The suitability of this signature peptide was tested for analytically representing the arginine kinase, by performing a multi-reaction monitoring tandem mass spectrometry approach on actual air filter samples collected from a simulated crab processing plant.