A scanning electron microscope study of a microsporidian with a pansporoblast: Thelohania maenadis

Air drying is not adequate for the preservation of the pansporoblastic membrane of Thelohania maenadis (Protozoa, Microsporidia), a parasite of the crabs Carcinus mediterraneus and Carcinus maenas. Freeze-drying and critical point drying preserve the pansporoblast membrane and reveal that isolated spores of the microsporidian are covered with a thick hairy coat. This coat originates as secretory product within the pansporoblast cavity.     

Shell shape affects movement patterns and microhabitat distribution in the hermit crabs Calcinus elegans, C. laevimanus and C. latens

This study examined the influence of shell shape on the distribution and movement patterns of three species of Hawaiian hermit crabs: Calcinus elegans, C. laevimanus, and C. latens. Field surveys showed strong differences in shell use depending on habitat. Individuals of C.elegans and C. latens were more frequently in unusual shapes of shells (the cowrie Cypraea caputserpentis and the variable worm shell Serpulorbis variabilis) when in tide pools and in more standard gastropod shells, such as the dog whelk Nassarius papillosus, when found in the subtidal. In addition, for both C.elegans and C. latens in tide pools, most crabs in unusual shaped shells were out on top of rocks, whereas most crabs in shells that were standard shapes were under rocks.In the laboratory, individuals of C.elegans and C. laevimanus in unusual shells initiated more shell exchanges and when given empty shells crabs readily occupied the standard shaped shells, but crabs did not move into the unusual shaped shells. Mark-recapture experiments in the field showed that C. elegans in standard shaped shells moved out of tide pools and stayed longer when placed on subtidal coral heads, whereas crabs in unusual shaped shells stayed in tide pools and did not stay on subtidal coral heads (in part due to predation). Laboratory tests showed that C. elegans in unusual shaped shells were more readily dislodged by surge than crabs in standard shaped shells. Thus, the difference in movement patterns in preferred vs. unpreferred shell shapes is an important factor influencing the microhabitat distribution of these hermit crabs.     

Position of horseshoe crabs in estuarine food webs: N and C stable isotopic study of foraging ranges and diet composition

To discern the position of horseshoe crabs as a potentially important predator in estuarine food webs, we determined where they foraged and what they ate. We used N and C stable isotopes to link adult horseshoe crabs to their oraging locations and potential food sources in Pleasant Bay, Cape Cod. The δ¹⁵N in tissues of horseshoe crabs and their potential foods suggest crabs were loyal to local foraging sites and did not forage substantially in subestuaries receiving >110 kg N ha⁻¹ year⁻¹. Among locations where crabs foraged, δ¹³C values in potential foods showed that food webs in subestuaries subject to higher N loads were supported by algal producers, while food webs in subestuaries with lower N loads were also supported by Spartina. δ¹³C values in horseshoe crab tissue did not change with load, suggesting they ate a mixed diet, regardless of N load. N and C isotopes in horseshoe crab feces were similar to signatures of estimated diet, suggesting low assimilation efficiency, perhaps due to ingestion of low quality organic matter. Although horseshoe crabs were relatively opportunistic in foraging habits, conservation or culture of horseshoe crabs may require habitats with higher water quality, ample particulate organic matter, and supporting a variety of prey.     

Hematodinium sp. infection of red Paralithodes camtschaticus and blue Paralithodes platypus king crabs from the Sea of Okhotsk, Russia

A disease caused by a parasitic dinoflagellate of the genus Hematodinium was identified in red, Paralithodes camtschaticus, and blue, Paralithodes platypus, king crabs from the north-east region of the Sea of Okhotsk, Russia, during annual stock surveys. No carapace color change was observed even in heavily infected crabs, but diseased crabs possessed creamy-yellow hemolymph, which was visible through the arthrodial membranes of the abdomen and appendages. Several stages of the parasite’s life history, including trophonts, plasmodia, sporonts and macrodinospores, were observed in tissues of infected king crabs. Numerous parasite cells were observed in the lumina of the myocardium, the gills, the connective tissue of antennal glands and the sinuses of nerve ganglia, eyestalks and gastrointestinal tract of king crabs with gross signs of infection. Based on sequencing of the 18S rDNA, it appears that the Hematodinium sp. found in red and blue king crabs is identical or closely related to Hematodinium sp. isolated from crabs of the genera Chionoecetes and Lithodes. Observed prevalences were 0.33% in sublegal male red king crabs, 0.18% in female red king crabs, 0.34% in sublegal male blue king crabs and 0.31% in female blue king crabs.     

Mitochondrial biogenesis during fungal spore germination. Development of cytochrome c oxidase activity.

Mitochondria from dormant spores of the fungus Botryodiplodia theobromae did not contain extractable cyctochrome c oxidase (EC 1.9.3.1) activity; however, this enzyme activity was elaborated rapidly after 150 min of the 240-min germination sequence. The absence of cytochrome c oxidase activity in the dormant spores apparently is not an artifact caused by spore disruption and fractionation procedures, transient enzyme instability, or insensitivity of the enzyme assay. Mitochondria from dormant spores of three other phylogenetically diverse genera of fungi were observed to contain readily detectable quantities of cytochrome c oxidase, suggesting that the absence of the enzyme in B. theobromae may be relatively novel. The elaboration of cytochrome c oxidase activity in germinating spores was abolished by cycloheximide if the drug was added at or before 95 min of germination, but development of enzyme activity was initially insensitive to inhibitors of the mitochondrial genetic system, chloramphenicol or ethidium bromide. Incubation of spores in both ethionine and S-2-aminoethyl-l-cysteine reduced the amount of extracted cytochrome c oxidase activity. Elaboration of enzyme activity was severely retarded by cerulenin, an inhibitor of fatty acid biosynthesis and of spore germination. This enzyme activity developed in water-incubated or 1% Tween 80-incubated spores in which only the cytoplasmic ribosomes are functional in translation of a stored nuclear messenger RNA. The results of this study show that cytoplasmic (but not mitochondrial) ribosome function is required for development of this enzyme activity during spore germination, and they suggest that a portion of the cytochrome c oxidase enzyme or some other protein required for its activity is synthesized de novo upon germination.     

Preference Alters Consumptive Effects of Predators: Top-Down Effects of a Native Crab on a System of Native and Introduced Prey

Top-down effects of predators in systems depend on the rate at which predators consume prey, and on predator preferences among available prey. In invaded communities, these parameters might be difficult to predict because ecological relationships are typically evolutionarily novel. We examined feeding rates and preferences of a crab native to the Pacific Northwest, Cancer productus, among four prey items: two invasive species of oyster drill (the marine whelks Urosalpinx cinerea and Ocenebra inornata) and two species of oyster (Crassostrea gigas and Ostrea lurida) that are also consumed by U. cinerea and O. inornata. This system is also characterized by intraguild predation because crabs are predators of drills and compete with them for prey (oysters). When only the oysters were offered, crabs did not express a preference and consumed approximately 9 juvenile oysters crab⁻¹ day⁻¹. We then tested whether crabs preferred adult drills of either U. cinerea or O. inornata, or juvenile oysters (C. gigas). While crabs consumed drills and oysters at approximately the same rate when only one type of prey was offered, they expressed a strong preference for juvenile oysters over drills when they were allowed to choose among the three prey items. This preference for oysters might negate the positive indirect effects that crabs have on oysters by crabs consuming drills (trophic cascade) because crabs have a large negative direct effect on oysters when crabs, oysters, and drills co-occur.     

Uptake of Bacteriophage and Their Subsequent Survival in Edible West Coast Crabs After Processing

Edible West Coast crabs (Cancer magister and Cancer antennarius), when in seawater contaminated with coliphage T4, were found to accumulate high titers of this virus. To study the extent of the hazard presented by crabs contaminated with virus, samples of edible crabs were contaminated with coliphage T4 and then processed by boiling. Results indicated that virus in crabs withstood this method of processing. The survival rate varied from 2.5 to 20%, depending upon the processing time. Heat penetration studies showed that, although internal temperature in the crabs was sufficient to inactivate virus, the processing times normally used to cook crabs were not. These results suggest that processed crabs could serve as vectors for the dissemination of virus diseases if the crabs are harvested from a polluted area.     

Chemically induced predator avoidance behaviour in the burrowing bivalve Macoma balthica

The responses of the burrowing bivalves Macoma balthica and Cerastoderma edule to chemical cues emitted by feeding shore crabs Carcinus maenas were investigated. M. balthica held in the laboratory and exposed to chemical signals in effluent water discharging from tanks containing C. maenas fed 20 M. balthica day^− 1 reacted by increasing their burial depths from approximately 30 mm to depths of > 60 mm, over a period of several days. When the signal was removed the bivalves gradually returned to their original depth over 5 days. C. edule similarly exposed to effluent from crabs feeding on conspecifics showed no response. In an attempt to identify the signal inducing this burrowing response, M. balthica were exposed to a variety of chemical signals. Crabs fed M. balthica elicited the strongest response, followed by crabs fed C. edule. There were also small responses to effluent from crabs fed on fish, crabs previously fed on M. balthica and to crab faeces, but no responses to starved crabs, crushed M. balthica, or controls. We conclude that increased burrowing depth of M. balthica is induced by some as yet unidentified chemical cue produced by feeding crabs and is strongest when the crabs were fed on M. balthica. Unexpectedly, neither the presence of crabs themselves, nor of damaged conspecifics, was effective in eliciting a burrowing response. The mortality rates of M. balthica and C. edule selected by crabs when burrowed at normal depths and after exposure to effluent from feeding crabs were different. Crabs selected 1.5 times more C. edule than M. balthica when both species were burrowed at their normal depths, but 15 times more after the tanks had been exposed to effluent from feeding crabs for 5 days. The burrowing response of M. balthica thus appears to reduce mortality significantly by displacing predation pressure on to the more accessible C. edule.     

Molluscan predation by Ovalipes punctatus (De Haan) (Crustacea: Brachyura: Portunidae)

Molluscan predation by the three-spot swimming crab was investigated. The dentition of the heteromorphic chelae allowed crushing, shearing, cutting and holding of prey. Laboratory investigations indicated that small mussels and gastropods were crushed, the larger mussels were prized open, and the foot of the larger gastropods shredded and bits removed. Stomach contents of freshly captured crabs indicated that the crabs are selective carnivores and preferred prey species which are not most abundant in situ (crabs from Kings Beach, Donax serra Röding; crabs from Maitlands River Beach, Bullia rhodostoma Reeve). Ovalipes punctatus (De Haan) foraged on a variety of prey and had no upper prey size limit, but the crabs did show preferences for certain prey sizes. Data indicate that the swimming crabs can effectively utilize the entire mollusc populations on the beaches as prey items.     

Cymo melanodactylus crabs slow progression of white syndrome lesions on corals

Predation on coral tissue by the crab Cymo melanodactylus has been hypothesized to contribute to tissue loss caused by white syndromes (WS) in acroporid corals. Here, we demonstrate that transplanting C. melanodactylus crabs from WS-infected Acropora colonies onto healthy coral fragments in controlled aquarium experiments does not result in WS transmission over a 21-day experimental period. Furthermore, progression of WS lesions was three times more rapid on corals with all C. melanodactylus crabs removed than on those with crabs (2.28 ± 0.21 vs. 0.74 ± 0.22 cm/day, respectively); thus, crabs slow WS disease progression under experimental conditions. In choice experiments, C. melanodactylus crabs were strongly attracted to corals with WS lesions, with 87 % of crabs migrating to WS fragments versus 3 % to healthy fragments. The strong attraction of C. melanodactylus to WS-infected corals and their ability to significantly reduce lesion progression rates suggest a mechanism whereby these coral-dwelling crabs could mitigate the effects of WS diseases on reefs.     

Hemolymph osmo-ionic concentrations in terrestrial crabs following desiccation

• 1.1. The hemolymph osmotic, Na⁺ K⁺, Ca⁺⁺ and Mg⁺⁺ concentrations were determined for both sexes in crabs from mangrove Ucides cordatus and Goniopsis cruentata and supralittoral Ocypode quadrala after aerial desiccation, for 10 hr.
• 2.2. There was no difference between sexes in the two mangrove crabs, but in O. quadrata the females were the most significantly affected (P < 0.01).
• 3.3. Hemolymph osmotic Na⁺, Ca⁺⁺ and Mg⁺⁺ concentrations increased significantly in desiccated ghost crabs, while K ⁺ concentration was not significantly changed. In the two mangrove crabs, only hemolymph Ca⁺⁺ concentration increased significantly, after desiccation.
• 4.4. The short time desiccation is suggested to be a device to study the steps of osmo-ionic regulation in terrestrial crabs.

     

Effect of Extracellular Products of Pseudoalteromonas atlantica on the Edible Crab Cancer pagurus

Previous studies have shown that injection of extracellular products (ECP) of Pseudoalteromononas atlantica isolated from shell disease-infected edible crabs (Cancer pagurus) into healthy crabs causes rapid death. In this study we examined the nature of the active lethal factor(s) in ECP. Injection of ECP into crabs caused a rapid decline in the total number of circulating hemocytes (blood cells), and the crabs died within 60 to 90 min. The individuals that died showed eyestalk retraction, limb paralysis, and lack of antennal sensitivity, suggesting that the active factor(s) targeted the nervous system. Histopathological investigations showed that affected crabs had large aggregates of hemocytes in the gills, and there was destruction of the tubules in the hepatopancreas. The active factor in ECP was not sensitive to heat treatment (100°C for 30 min) and proteinase K digestion. As lipopolysaccharide (LPS) was a potential candidate for the lethal factor, it was purified from whole P. atlantica bacteria or ECP and subsequently injected into crabs. These crabs had all of the external symptoms observed previously with ECP, such as limb paralysis and eyestalk retraction, and they died within 90 min after challenge, although no significant decline in the number of circulating hemocytes was observed. Similarly, in vitro incubation of hemocytes with purified LPS (1 to 20 μg) from P. atlantica did not result in the clumping reaction observed with ECP but did result in a degranulation reaction and eventual cell lysis. Injection of crabs with Escherichia coli or Pseudomonas aeruginosa LPS (1 μg g of body weight⁻¹) did not cause any of the characteristic symptoms observed following exposure to P. atlantica LPS. No mortality of crabs followed the injection of E. coli LPS, but P. aeruginosa LPS caused ca. 80% mortality at 2 h after injection. Overall, these results show that the main virulence factor of P. atlantica for edible crabs is LPS either alone or in combination with other heat-stable factors.     

Identification and Characterization of Two Subpopulations of Encephalitozoon intestinalis

Microsporidia are obligate intracellular protozoa that have been shown to be pathogenic to most living creatures. The development of in vitro cell culture propagation methods has provided researchers with large numbers of spores and facilitated the study of these organisms. Here, we describe heterogeneity within cell culture-propagated Encephalitozoon intestinalis suspensions. Flow cytometer histograms depicting the log side scatter and forward-angle light scatter of spores from nine suspensions produced over 12 months consistently showed two populations differing in size. The suspensions were composed primarily of the smaller-spore subpopulation (76.4% ± 5.1%). The presence of two subpopulations was confirmed by microscopic examination and image analysis (P < 0.001). Small subpopulation spores were noninfectious in rabbit kidney (RK13) cell culture infectivity assays, while the large spores were infectious when inocula included ≥25 spores. The small spores stained brilliantly with fluorescein isothiocyanate-conjugated monoclonal antibody against Encephalitozoon genus spore wall antigen, while the large spores stained poorly. There was no difference in staining intensities using commercial (MicroSporFA) and experimental polyclonal antibodies. Vital-dye (DAPI [4′,6′-diamidino-2-phenylindole], propidium iodide, or SYTOX Green) staining showed the spores of the small subpopulation to be permeable to all vital dyes tested, while spores of the large subpopulation were not permeable in the absence of ethanol pretreatment. PCR using primers directed to the 16S rRNA or β-tubulin genes and subsequent sequence analysis confirmed both subpopulations as E. intestinalis. Our data suggest that existing cell culture propagation methods produce two types of spores differing in infectivity, and the presence of these noninfective spores in purified spore suspensions should be considered when designing disinfection and drug treatment studies.     

Brittle-star bioluminescence functions as an aposematic signal to deter crustacean predators

The behaviour of three species of nocturnally active, visually orienting crabs (Portunus sebae, P. spinnimanus and P. ordwayi) was observed to determine whether the luminescent signals produced by Ophiopsila riisei (Echinodermata: Ophiuroidea) function as an aposematic deterrent against crustacean predators. In repeated experimental trials, crabs showed more rapid rejection of luminescent unpalatable ophiuroids than of non-luminescent controls. After five trials, unpalatable luminescent prey were rejected three times as quickly as either unpalatable or palatable non-luminescent controls. Over the course of the trials, crabs damaged fewer luminescent ophiuroids than non-luminescent controls. Furthermore, blind crabs (that could not perceive the luminescent flashes) caused significantly more damage to luminescent brittle-stars than did crabs with intact eyes (that could see the light signals). All three species of crab ophiuroids versus non-luminescent controls. Ophiopsila riisei survived the majority of crab attacks, and crabs can therefore learn to reject unpalatable luminescent prey without killing them. This suggests that individual selection may be an important mechanism for the evolution of aposematic luminescent signals in this, and probably other, luminescent species.     

Predator-prey dynamics between recently established stone crabs (Menippe spp.) and oyster prey (Crassostrea virginica)

Range expansion and population establishment of individual species can have significant impacts on previously established food webs and predator-prey dynamics. The stone crab (Menippe spp.) is found throughout southwestern North Atlantic waters, from North Carolina through the Gulf of Mexico and the Central American Caribbean, including the Greater Antilles. Recent observations suggest that stone crabs have become better established on certain oyster reefs in North Carolina than in the early 1900s when they we first observed in NC. To assess the predatory impact of stone crabs on oysters, we (1) quantified stone crab densities on subtidal oyster reefs in Pamlico Sound, NC using scuba surveys, and (2) conducted laboratory predation experiments to assess the functional response of stone crabs to varying densities of oysters. We then (3) analyzed previously unpublished functional response data on another important oyster predator, the mud crab Panopeus herbstii. Finally, we (4) compared and contrasted potential predatory impacts of stone, mud and blue crabs (Callinectes sapidus). The functional response data and analyses for both stone crabs and mud crabs were consistent with a type II functional response. Mud crabs, on a m² basis, inflicted the highest proportional mortality on oysters over a 24 hour period, followed by stone and then blue crabs. Proportional mortality did not vary significantly with oyster size; however, relatively small and large oysters were consumed disproportionately less than medium-sized oysters, likely due to the mechanical inability of stone crabs to handle small oysters, and the inability to crush large oysters. Although stone crabs appear to be established in Pamlico Sound at densities equivalent to densities in other systems such as the U.S. Florida Panhandle, their predatory activities on oysters are not expected to have as significant a negative impact on oyster populations compared to other resident predators such as mud crabs.     

Predatory behavior and preference of a successful invader, the mud crab Dyspanopeus sayi (Panopeidae), on its bivalve prey

Predator-prey relationships between the panopeid crab, Dyspanopeus sayi, and the mytilid, Musculista senhousia, were investigated. Through laboratory experiments, prey-handling behavior, prey size selection, predator foraging behavior and preferences for two types of prey (M. senhousia and the Manila clam Ruditapes philippinarum) were assessed. Handling time differed significantly with respect to the three prey sizes offered (small: 15.0-20.0 mm shell length, SL; medium: 20.1-25.0 mm SL; and large: 25.1-30.0 mm SL); mud crabs were more efficient in predating medium-small than large prey. Although differences in prey profitability were not evident, D. sayi exhibited a marked reluctance to feed on larger-sized prey whilst smaller, more easily predated mussels were available. Size selection may be the result of a mechanical process in which encountered prey are attacked but rejected if they remain unbroken after a certain number of opening attempts. D. sayi exhibited inverse density-dependent foraging. A significant higher mortality of prey was evident at low prey density. Thus, at low predator density, the D. sayi-M. senhousia interaction was a destabilizing type II functional response. Interference responses affected the magnitude of predation intensity by D. sayi on M. senhousia, since as the density of foraging crabs increased, their foraging success fell. At high density (4 crabs tank⁻¹), crabs engaged in a high amount of agonistic activity when encountering a conspecific specimen, greatly diminished prey mortality. Finally, presenting two types of prey, Manila clam juveniles were poorly predated by mud crabs, which focused their predation mostly on M. senhousia. It is hypothesized that, when more accessible prey is available, mud crabs will have a minimal predatory impact on commercial R. philippinarum juvenile stocks.     

Effect of gastropod shell characteristics and hermit crabs on shell Epifauna

The epifauna on gastropod shells occupied by the hermit crabs Pagurus pollicaris (Say) and P. longicarpus (Say) was examined, as was the utilization of shells by these two hermit crabs. In the study area in Tampa Bay, Florida, shells were not a limiting factor to the hermit crab population, and there apparently was little competition for shells. Interspecific competition for shells was limited because the two hermit crab species differed in size and hence occupied shells of different sizes. The total number and density of most epifaunal species were higher on shells occupied by hermit crabs than on unoccupied shells, possibly because hermit crabs prevent their shells from being buried and hence lengthen the time the epifaunal community can grow and develop. The hermit crab species also appeared to affect the epifaunal community, for the total number and density of most epifaunal species were larger on shells occupied by P. pollicaris than P. longicarpus. With increasing shell size, the populations of most epifaunal species, also were larger but not their density. Least influential in affecting the epifaunal community was the species of shells.     

A forced association between the slippersnail Crepidula convexa and the hermit crab Pagurus longicarpus?—possible influence from a third party

The suspension-feeding slippersnail Crepidula convexa is commonly associated with hermit crabs (Pagurus longicarpus) living in periwinkle shells (Littorina littorea) at our study site in Nahant, MA, USA. In 15 field surveys conducted at Nahant in 2000, 2001 and 2003, we found that (1) more than 61.8% of individuals of C. convexa resided on shells occupied by hermit crabs, as opposed to the shells of live periwinkles, empty periwinkle shells or other solid substrates; (2) an average of 8.3% of hermit crabs carried at least one individual of C. convexa; and (3) 39.1-75.0% of hermit crabs carrying C. convexa were carrying “large” individuals (snails with wet weight >10% of the weight of the periwinkle shells they occupied). However, it is unlikely that individuals of C. convexa seek out shells occupied by hermit crabs to colonize, and they showed no preference for empty periwinkle shells over other solid substrates in the laboratory. Moreover, in the laboratory the hermit crabs preferentially occupied intact shells bearing individuals of C. convexa only when the alternatives were shells that had been drilled by naticid snails. Thus, neither party preferentially associates with the other: rather, extensive predation by naticid snails on periwinkles at Nahant appears to limit the availability of suitable shells for the hermit crabs, forcing them to inhabit shells bearing “large” individuals of C. convexa. Individuals of C. convexa may benefit from this inadvertent association with hermit crabs: by facilitating snail dispersal, transport by hermit crabs should reduce the potential for inbreeding, an important consideration for a species that lacks free-living larvae in its life history.     

Risk of Handling as a Route of Exposure to Infectious Waterborne Cryptosporidium parvum Oocysts via Atlantic Blue Crabs (Callinectes sapidus)

Commercial Atlantic blue crabs (Callinectes sapidus) were exposed to 2.0 × 10⁴ infectious waterborne oocysts of Cryptosporidium parvum. The study demonstrated that blue crabs can transfer C. parvum oocysts to persons involved in handling or preparing crabs and that they may contaminate other surfaces or products during storage.     

Killing of Bacillus subtilis Spores by a Modified Fenton Reagent Containing CuCl2 and Ascorbic Acid

Bacillus subtilis spores were killed by CuCl₂-ascorbic acid, chloride ions were essential for killing of spores, and spores with defective coats were killed more rapidly. CuCl₂-ascorbic acid did not damage spore DNA, and spores killed by this reagent initiated germination. However, spores killed by CuCl₂-ascorbic acid may have damage to their inner membrane.