Tissue changes in the shore crab Carcinus maenas as a result of infection by the parasitic barnacle Sacculina carcini

We studied the effect of parasitic invasion by the barnacle Sacculina carcini on shore crabs Carcinus maenas collected from the Menai Straits in North Wales, UK. A significant reduction in serum protein and circulating granular, semi-granular and hyaline blood cells was observed in parasitised individuals, while serum ammonium and glucose concentrations were significantly increased. No difference in hepatopancreatic glycogen concentration was found between healthy and parasitised crabs. Histological analysis showed the apparent removal of fibrillar protein from infected muscle by the parasite. Hepatopancreas tubule necrosis was also routinely observed in infected individuals. Parasitisation by S. carcini dramatically affects the haemocyte population and serum chemistry of infected crabs.     

A pheromone bouquet controls the reproductive behaviour of the male shore crab,Carcinus maenas

Aquatic Ecology - The reproduction of many brachyuran crustaceans involves the formation of mating pairs often around the time of the female moult with attraction of a sexual partner and mating...     

Sex-specific mediation of foraging in the shore crab, Carcinus maenas

Experiments were conducted to investigate the sex-specific differences to feeding responses of the shore crab Carcinus maenas throughout the year. Results demonstrate that female shore crabs exhibit stronger feeding responses than males throughout the year with a significantly reduced feeding response in males during the summer months' reproductive season. We also studied the possible function(s) of the moulting hormone, 20-hydroxyecdysone (Crustecdysone) that has been described as a potential female-produced sex pheromone to initiate male reproductive behaviour in a number of crustaceans. We recently presented evidence that for shore crabs this is not the case and now show that the steroid is instead functioning as a sex-specific feeding deterrent protecting the moulting 'soft' female crabs. Whilst male shore crabs were deterred from prey (Mytilus edulis) and synthetic feeding stimulants glycine and taurine when these feeding stimulants were spiked with crustecdysone, intermoult female crabs were significantly less affected and rarely deterred from feeding. This sex specificity of the moulting hormone, in combination with the female sex pheromone, which has no anti-feeding properties, ensures that male crabs mate with soft-shelled, moulted females rather than engage in cannibalism, such as found frequently in cases when soft-shelled females are exposed to intermoult females.     

The effect of dietary chitin supplementation on the survival and immune reactivity of the shore crab, Carcinus maenas

Adult male shore crabs (Carcinus maenas) were maintained on a fish-based diet supplemented with 0, 5 or 10% chitin for 11 weeks. Significantly greater mortality was found during this period in the control group (0% chitin) than those fed 10% chitin. Crabs fed 5 or 10% chitin had lower numbers of cultivatable bacteria in the hepatopancreas than those on the basal diet alone. The addition of chitin had no significant effect on the serum concentrations of protein and glucose, and the levels of glycogen in the hepatopancreas. The total number of circulating hemocytes in the blood was unaffected by the addition of chitin to the diet, however, at week 6 there were significantly more hyaline hemocytes in those crabs fed 10% chitin than the control group. The in vitro phagocytic activity of hemocytes was unaffected by chitin supplementation and crabs challenged with Vibrio alginolyticus showed a similar pattern of susceptibility in the three dietary groups (0, 5 or 10% chitin). Overall although crabs on a chitin-supplemented diet showed greater survival, this was not explained in terms of elevation in the cellular defences of these animals. The enhanced survival of crabs-fed chitin is probably as a result of the removal of potentially pathogenic bacteria from the hepatopancreas. Because chitin appears to 'purge' bacteria from the gut, this may prove to be a useful addition to diets on animals undergoing oral probiotic treatment.     

Developing the options for managing marine pests: specificity trials on the parasitic castrator, Sacculina carcini, against the European crab, Carcinus maenas, and related species

The impacts of introduced marine pests are becoming increasingly apparent, prompting interest in the possibility of their biological control. We undertook laboratory and field experiments on host selection of one potential control agent (the endoparasitic barnacle, Sacculina carcini) against its natural host (the widely invasive European shore crab, Carcinus maenas) and several confamilial and more distantly related crustaceans. For comparison, we also tested host specificity in a related parasitic barnacle, Heterosaccus lunatus. The results confirm indistinct behavioral host selection in S. carcini, indicate very different mechanisms for host selection by S. carcini and H. lunatus (which could be related to differences between the two species in attachment points), and suggest host specificity in S. carcini depends on interactions between the parasite and the host's physiology. Development of convincing safety trials for marine parasites like S. carcini, in which the infective stage is a planktonic larva, will be more difficult than for many terrestrial parasites and will require detailed knowledge of the parasite's behavior and physiological interaction with its hosts.     

A membrane-bound hemoglobin from gills of the green shore crab Carcinus maenas

Most hemoglobins serve for the transport or storage of O(2). Although hemoglobins are widespread in "entomostracan" Crustacea, malacostracans harbor the copper-containing hemocyanin in their hemolymph. Usually, only one type of respiratory protein occurs within a single species. Here, we report the identification of a hemoglobin of the shore crab Carcinus maenas (Malacostraca, Brachyura). In contrast to the dodecameric hemocyanin of this species, C. maenas hemoglobin does not reside in the hemolymph but is restricted to the gills. Immunofluorescence studies and cell fractioning showed that C. maenas hemoglobin resides in the membrane of the chief cells of the gill. To the best of our knowledge, this is the first time that a membrane-bound hemoglobin has been identified in eukaryotes. Bioinformatic evaluation suggests that C. maenas hemoglobin is anchored in the membrane by N-myristoylation. Recombinant C. maenas hemoglobin has a hexacoordinate binding scheme at the Fe(2+) and an oxygen affinity of P(50) = 0.5 Torr. A rapid autoxidation rate precludes a function as oxygen carrier. We rather speculate that, analogous to prokaryotic membrane-globins, C. maenas hemoglobin carries out enzymatic functions to protect the lipids in cell membrane from reactive oxygen species. Sequence comparisons and phylogenetic studies suggested that the ancestral arthropod hemoglobin was most likely an N-myristoylated protein that did not have an O(2) supply function. True respiratory hemoglobins of arthropods, however, evolved independently in chironomid midges and branchiopod crustaceans.     

No evidence of morphine analgesia to noxious shock in the shore crab, Carcinus maenas

A number of criteria have been suggested for testing if pain occurs in animals, and these include an analgesic effect of opiates (Bateson, 1991). Morphine reduces responses to noxious stimuli in crustaceans but also reduces responsiveness in a non-pain context. Here we use a paradigm in which shore crabs receive a shock in a preferred dark shelter but not if they remain in an unpreferred light area. Analgesia should thus enhance movement to the preferred dark area because they should not experience ‘pain’. However, morphine inhibits rather than enhances this movement even when no shock is given. Morphine produces a general effect of non-responsiveness rather than a specific analgesic effect and this could also explain previous studies claiming analgesia. However, we question the utility of this criterion of pain and suggest instead that behavioural criteria be employed.     

Haemolymph parasite of the shore crab Carcinus maenas: pathology, ultrastructure and observations on crustacean haplosporidians

A protozoan parasite with some features of haplosporidians is described from the European shore crab Carcinus maenas. The parasite establishes a systemic infection through the haemal sinuses and connective tissues. Intracellular stages of the parasite were found within reserve inclusion, connective tissue, and muscle cells, while free forms were present in all haemal spaces. A uninucleate stage appeared to develop to a multinucleate plasmodial stage following multiple mitotic divisions of the nucleus. Histopathology also indicated that nuclear division may occur to form multinucleate plasmodia, in connective tissue, reserve inclusion and muscle cells, the multinucleate plasmodium being enclosed in the host-cell plasma membrane. It appears that the multinucleate plasmodium may then undergo internal cleavages which result in plasmodial fragmentation to form many uninucleate stages. Both stages, but particularly the uninucleate stage, contained cytoplasmic, large, ovoid, dense vesicles (DVs), some of which contained an internal membrane separating the medulla from the cortex, as in haplosporosomes. Golgi-like cisternae, closely associated with the nuclear membrane, formed DVs and haplosporosome-like bodies (HLBs), superficially resembling viruses. Infrequently, HLBs may condense to form haplosporosomes. The DVs, as in spores of some Haplosporidium spp. and paramyxeans, may give rise to, and are homologous with, haplosporosomes. Other features, such as the presence of an intranuclear mitotic spindle, lipid droplets, and attachment of DVs and haplosporosomes to the nuclear membrane, indicate that the C. maenas parasite is a haplosporidian. A similar organism reported from the haemolymph of spot prawns Pandalus spp., and haplosporidians reported from prawns Penaeus vannamei and crabs Callinectes sapidus may belong to this group. It is concluded that the well-characterised haplosporidians of molluscs and some other invertebrates may not be characteristic of the whole phylum, and that morphologically and developmentally similar organisms may also be haplosporidians, whether they have haplosporosomes or not.     

Sex pheromone in the shore crab Carcinus maenas,and the site of its release from females

Sex pheromones are demonstrated in premoult and recently‐moulted female Carcinus maenas (L.) and Macropipus holsatus (Fabr.) (Crustacea Decapoda, Portunidae). Male conspecifics respond with searching activity. Dilute solutions of urine aspirated from the antennal glands of females of each species release characteristic responses in conspecific males, but males of other species respond differently, if at all. Aspects of responses of test animals in bioassays are discussed.     

Fine structure and morphogenesis of the sclerite epicuticle in the Atlantic shore crab Carcinus maenas

Three basic sublayers are identified in the epicuticle of the mineralised sclerites of the Atlantic shore crab Carcinus maenas (Crustacea, Decapoda): the surface coat, the cuticulin layer, and the inner epicuticle. Their morphogenesis and subsequent changes are described throughout the moulting cycle in the normal cuticle and the cuticular structures, namely the sensory bristles and epicuticular spines. At first, the cuticulin layer begins to form just after apolysis. This layer is built directly over the plasma membrane and immediately appears as a membrane-like structure 40 nm thick, composed of five symmetrically arranged laminae: two inner electron-lucent leaflets sandwiched between two thick electron-dense leaflets and separated by a thin dense median stratum. Elaboration of the inner epicuticle below the cuticulin layer is thought to occur via an intussusceptive process involving the pore canal cell extensions as transport routes. The inner epicuticle is made of vertically oriented microfibres embedded in an electron-dense matrix material. During the second half of the premoult period, the surface coat is deposited on the upper side of the cuticulin layer.     

Effects of mercury on ion and osmoregulation in the shore crab Carcinus maenas (L.)

The effects of lethal mercury concentrations on regulation of hemolymph electrolytes in the shore crab Carcinus maenas were investigated. In most experiments mercury exposure reduced hemolymph osmolality and Na+, Cl- and K+ levels to 55-90% of controls in 48 hr. Mercury exposure augmented calcium levels to 120-300% of controls. The effect of mercury on all of the electrolytes varied during the year, especially on magnesium levels which showed significant increases or decreases, or remained unaffected in different experiments.     

Neural control of ventilation in the shore crab, Carcinus maenas. II. Frequency-modulating interneurons

1. We have identified a class of nonspiking interneurons which can control the frequency of ventilation in a graded manner. These frequency modulating interneurons (FMis) also receive synaptic inputs in-phase with the ventilatory motor output providing a functional positive feedback loop in the ventilatory system. The class of FMis is composed of three morphologically and physiologically distinct interneurons, FMi1, FMi2 and FMi3. 2. Depolarization of FMi1 increases the rate of ventilation, while hyperpolarization decreases the rate (Fig. 1). This control is restricted to a single ventilatory central pattern generator (CPG), (Fig. 2), although FMi1 sends processes into the neuropils of both hemiganglionic CPGs (Fig. 3). 3. Hyperpolarization of FMi2 increases the rate of both ventilatory CPGs while depolarization of this cell slows and eventually arrests the rhythm (Figs. 5 and 6). FMi2 receives a synaptic input correlated with the motor output of each of the ventilatory CPGs (Fig. 4). During periods of reversed ventilation, this cell is abruptly hyperpolarized and continues to be driven in-phase with the ventilatory motor output (Fig. 7). 4. Hyperpolarization of FMi3 increases the rate of ventilation and depolarization decreases the rate of ventilation produced by both CPGs (Fig. 10). This control of the ventilatory rate by FMi3 is graded (Fig. 11). There is no apparent change in the membrane potential of FMi3 during reversed ventilation and it is morphologically distinct from FMi2. 5. FMi2 and FMi3 may be involved in the switch in ventilatory motor pattern from forward to reversed ventilation. Hyperpolarization of FMi2 and depolarization of FMi3 can elicit bouts of reversed ventilation from both CPGs (Fig. 13). 6. These results suggest that the FM interneurons act in parallel to control the frequency of ventilation and may act as integrating elements between spiking 'command' fibers in the circumesophageal connectives and the nonspiking interneurons of the ventilatory CPG.     

Organization of a phyllobranchiate gill from the green shore crab Carcinus maenas (Crustacea,Decapoda)

Summary The phyllobranchiate gills of the green shore crab Carcinus maenas have been examined histologically and ultrastructurally. Each gill lamella is bounded by a chitinous cuticle. The apical surface of the branchial epithelium contacts this cuticle, and a basal lamina segregates the epithelium from an intralamellar hemocoel. In animals acclimated to normal sea water, five epithelial cell types can be identified in the lamellae of the posterior gills: chief cells, striated cells, pillar cells, nephrocytes, and glycocytes. Chief cells are the predominant cells in the branchial epithelium. They are squamous or low cuboidal and likely play a role in respiration. Striated cells, which are probably involved in ionoregulation, are also squamous or low cuboidal. Basal folds of the striated cells contain mitochondria and interdigitate with the bodies and processes of adjacent cells. Pillar cells span the hemocoel to link the proximal and distal sides of a lamella. Nephrocytes are large, spherical cells with voluminous vacuoles. They are rimmed by foot processes or pedicels and frequently associate with the pillar cells. Glycocytes are pleomorphic cells packed with glycogen granules and multigranular rosettes. The glycocytes often mingle with the nephrocytes. Inclusion of the nephrocytes and glycocytes as members of the branchial epithelium is justified by their participation in intercellular junctions and their position internal to the epithelial basal lamina.     

Corrective note about R* cells of the digestive gland of the shore crab Carcinus maenas

In a previous paper, we described and discussed the possible functions of calcospherite-rich cells (R^* cells) in the digestive gland of the shore crab, Carcinus maenas. We recently realised that electron micrographs in this publication presented neither typical R^* cells nor their calcium phosphate granules. Indeed, our pictures showed spermatophores (filled with typical spermatozoa) that had contamined hepatopancreatic cell suspensions. As the present study indicates, this contamination is difficult to detect by optical microscopy because unstained R^* cells closely resemble spermatophores. However, morphological differences between these cell types appear clearly when observed by electron microscopy. The present paper describes a comparative study of cell populations isolated from female digestive glands; it validates our previous results obtained with male hepatopancreas and suggests a low containation of those male cell fractions by spermatophores.     

On the fine structure of the regressing ecdysial glands of Carcinus maenas L. (Crustacea decapoda) parasitized by Sacculina carcini thompson

Summary The ecdysial glands (Y organs) of the crab Carcinus maenas regress in the presence of an external parasite, Sacculina carcini. This regression is more or less severe and may lead to complete autolysis. Three gradual stages in this involutionary process are described. In stage I, the gland cells are nearly normal. Nuclei and cytoplasmic organelles remain unchanged, but large vacuoles begin to appear. Stage II corresponds to more or less drastic nuclear pyknosis and cytoplasmic alterations. Myelin figures are large and numerous. Lysosomes and autophagic vacuoles with phosphatase activity are abundant. However, the general cellular architecture remains preserved. Stage III corresponds to irreversible cytolysis; nuclear envelopes and plasma membranes have disappeared. What remains is an accumulation of cellular debris becoming engulfed by circulating hemocytes. Not all of the gland cells of any given Y organ show the same degree of regression; degeneration is asynchronous.Structures seemingly corresponding to absorptive roots of the parasite are seen. Their lumen is coated with microvilli. The putative direct and indirect influences of the rhizocephalan parasite on its host are discussed. Our results on regressing Y organs of parasitized crabs are compared with those on regressing ecdysial glands of insects.Dedicated to the memory of Sir Francis Knowles, the first investigator to examine the ultrastructure of the Y organ of Carcinus maenas We wish to express our thanks to Professor Berta Scharrer for her critical advice