Parasites in the thoracic ganglion of Pachygrapsus marmoratus (Brachyura: Grapsidae) from the coast of Portugal

We examined 149 marbled shore crabs, Pachygrapsus marmoratus, from the coast of Portugal for parasites. In particular, we focused our effort on the crab thoracic ganglion. The thoracic ganglion is the largest concentration of nervous tissue in a crab and thus, parasites associated with this organ are well situated to influence host behavior. We found metacercariae of two microphallid trematode species in the thoracic ganglion. We also found a microsporan and an apicomplexan associated with the thoracic ganglion. Other parasites not associated with the thoracic ganglion included gregarine trophozoites which were present in the digestive diverticulae in some of the crabs and the entoniscid isopod, Grapsion cavolini. Metacercariae of one of the trematodes (probably Microphallus pachygrapsi (Deblock and Prevot)), may influence the mortality of its host.     

Black mat syndrome,an invasive mycotic disease of the tanner crab, Chionoecetes bairdi

Black mat syndrome, caused by an encrusting fungus on the exterior of the carapace of tanner crabs, has been known for many years. Although it is a nuisance when processing crabs for the commercial market, it has been thought to be restricted to the external surfaces of the crab and, therefore, nonpathogenic. In the present study, 11 tanner crabs from the Kodiak area of Alaska with and 9 without grossly recognizable masses of the fungus on the carapace were necropsied and examined histologically utilizing special stains selective for fungi. In all individuals with the syndrome, hyphae of the fungus, previously identified as Phoma fimeti, penetrated the carapace and virtually replaced the underlying epidermis. In the more advanced cases, the eyestalk was invaded and the epidermis destroyed, and hyphae extended into the eyestalk musculature and nervous tissue. To date, infections of the connective tissue sheaths surrounding the esophagus, stomach, heart, hemopoetic tissue, thoracic ganglion, antennal gland, and ovary have also been observed. None of the crabs without the syndrome contained internal hyphae. Although data on the lethality of the disease are not yet available, the ease with which the hyphae penetrate the chitinous exoskeleton, their extensive proliferation in the epidermis, and their ability to invade deep tissues causing obvious pathological effects, are highly suggestive that it is a virulent, probably fatal, disease that may have a significant impact on tanner crab population dynamics.     

Clonal diversity of the marine trematode Maritrema novaezealandensis within intermediate hosts: the molecular ecology of parasite life cycles

We quantified the clonal diversity of the New Zealand marine trematode Maritrema novaezealandensis (n = 1250) within Zeacumantus subcarinatus snail (n = 25) and Macrophthalmus hirtipes crab (n = 25) intermediate hosts using four to six microsatellite loci, and investigated the potential biological and physical factors responsible for the observed genetic patterns. Individual snails harboured one to five trematode genotypes and 48% of snails were infected by multiple parasite genotypes. Overall, the number of parasite genotypes did not increase with snail size, but was highest in intermediate-sized snails. Significantly larger numbers of parasite genotypes were detected in crabs (relative to snails; P < 0.001), with 16-25 genotypes recovered from individual crabs. Although crabs are typically infected by small numbers of cercariae sourced from many snails, they are occasionally infected by large numbers of cercariae sourced from single snails. The latter cases explain the significant genetic differentiation of trematode populations detected among their crab hosts (F(ST) = 0.009, P < 0.001). Our results suggest that the timing of infection and/or intraspecific competition among parasite clones within snails determine(s) the diversity of parasite clones that snails harbour. The presence of a large number of infected snails and tidal mixing of cercariae prior to infection results in crabs potentially harbouring hundreds of parasite genotypes despite the crabs' territorial behaviour.     

Tyrosine hydroxylase protein expression in ventral nerve cord of Neotropical freshwater crab

Given the importance of catecholamines in coordinating physiological and behavioral responses in brachyurans, the present study was designed to investigate the distribution of tyrosine hydroxylase (TH)-positive cells and fibers in the ventral nerve cord of Dilocarcinus pagei the Neotropical freshwater crab. TH immunoreactivity was visualized in adult crabs of both sexes, during the intermolt period. We found TH-positive cells that have not been previously described in brachyurans. Specifically, we found a pair of TH-positive cells in the ventral region of the thoracic ganglion, and in ventral and dorsal regions of the abdominal (pleonic) ganglion, suggesting catecholaminergic modulation of claws’ function and abdominal structures. In addition, great population of TH-positive cells was observed in the subesophageal ganglion, indicating conservation during evolution of catecholamines in this ganglion of decapods. Dopamine is present in cells and fiber tracts of brachyuran ventral nerve cord, projecting to endocrine, cardiac and digestive structures, suggesting widespread modulation and control of physiological functions and behavior. Dopamine plays a central role in movement and psychiatric disorders in humans. Information on dopaminergic function in the nervous system of invertebrates should improve the understanding of its function in more complex systems, such as human beings.     

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.     

Selective crab predation on native and introduced bivalves in British Columbia

Experiments were conducted to determine whether locally abundant crab species prefer co-occurring littleneck clams, Protothaca staminea (Conrad, 1837) and Tapes philippinarum (A. Adams and Reeve, 1850), relative to a recently introduced species, the varnish clam, Nuttallia obscurata, (Reeve, 1857). Prey preference, handling time, pick-up success, profitability and consumption rates were investigated for two crab species, Dungeness crab, Cancer magister (Dana, 1852) and red rock crab, Cancer productus (Randall, 1839) crabs. Both crab species preferred varnish clams over the native species. This may be attributable to the lower handling time, higher pick-up success and increased profitability of consuming varnish clams. Handling time appeared to be a factor not only in species preference, but also in the degree of preference, with shorter handling times corresponding to stronger preference values. Both native and introduced bivalves burrow into the substratum, with the varnish clam burrowing deepest. When feeding on clams in limited substratum both crab species preferred the varnish clam. In the unlimited substratum trials Dungeness crabs preferred varnish clams (although to a lesser degree) while red rock crabs preferred littleneck clams. This was likely due to the significantly deeper burial of the varnish clam, making it less accessible. Although the morphology (i.e. thin shell, compressed shape) of the invader increases its vulnerability to predation, burial depth provides a predation refuge. These results demonstrate how interactions between native predators and the physical characteristics and behaviour of the invader can be instrumental in influencing the success of an invasive species.     

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.     

Molecular Cloning and Gene Expression Analysis of the Leptin Receptor in the Chinese Mitten Crab Eriocheir sinensis

Background

Leptin is an adipocyte-derived hormone with multiple functions that regulates energy homeostasis and reproductive functions. Increased knowledge of leptin receptor function will enhance our understanding of the physiological roles of leptin in animals.

Methodology/Principal Findings

In the present study, a full-length leptin receptor (lepr) cDNA, consisting of 1,353 nucleotides, was cloned from Chinese mitten crab (Eriocheir sinensis) using rapid amplification of cDNA ends (RACE) following the identification of a single expressed sequence tag (EST) clone in a cDNA library. The lepr cDNA consisted of a 22-nucleotide 5′-untranslated region (5′ UTR), a 402-nucleotide open reading frame (ORF) and a 929-nucleotide 3′ UTR. Multiple sequence alignments revealed that Chinese mitten crab lepr shared a conserved vacuolar protein sorting 55 (Vps55) domain with other species. Chinese mitten crab lepr expression was determined in various tissues and at three different reproductive stages using quantitative real-time RT-PCR. Lepr expression was highest in the intestine, thoracic ganglia, gonad, and accessory gonad, moderate in hepatopancreas and cranial ganglia, and low in muscle, gill, heart, haemocytes, and stomach. Furthermore, lepr expression was significantly higher in the intestine, gonad and thoracic ganglia in immature crabs relative to precocious and mature crabs. In contrast, lepr expression was significantly lower in the hepatopancreas of immature crabs relative to mature crabs.

Conclusions/Significance

We are the first to identify the lepr gene and to determine its gene expression patterns in various tissues and at three different reproductive stages in Chinese mitten crab. Taken together, our results suggest that lepr may be involved in the nutritional regulation of metabolism and reproduction in Chinese mitten crabs.     

An experimental evaluation of host specificity: the role of encounter and compatibility filters for a rhizocephalan parasite of crabs

The encounter/compatibility paradigm of host specificity provides three qualitative pathways to the success or failure of a potential host-parasite interaction. It is usually impossible to distinguish between two of these (encounter and compatibility filters closed versus encounter filter open and compatibility filter closed) because unsuccessful infection attempts are difficult to observe in nature. We were able to open the encounter filter under experimental laboratory conditions. Our analytical system used the rhizocephalan barnacle, Sacculina carcini, a parasitic castrator of the European green crab, Carcinus maenas, and Pachygrapsus marmoratus, a native European crab that occurs with C. maenas but is not parasitized by S. carcini in nature. Penetration followed by unsuccessful infection of P. marmoratus crabs by parasitic barnacle larvae leaves a uniquely permanent record in the thoracic ganglion of the crabs. This provided us with a novel tool to quantify the encounter filter in a host-parasite system in nature. We demonstrated, in the laboratory, that the compatibility filter was closed and that, in nature, even where barnacle larvae were present, the encounter filter was also effectively closed. The closure of both filters in nature explains the failure of this potential host-parasite interaction, an outcome favored by selection in both host and parasite.     

Distribution and characterization of Corazonin in the central nervous system of Triatoma infestans (Insecta: Heteroptera)

The distribution of corazonin in the central nervous system of the heteropteran insect Triatoma infestans was studied by immunohistochemistry. The presence of corazonin isoforms was investigated using MALDI-TOF mass spectrometry in samples containing the brain, the subesophageal ganglion, the corpora cardiaca-corpus allatum complex and the anterior part of the aorta. Several groups of immunopositive perikarya were detected in the brain, the subesophageal ganglion and the thoracic ganglia. Regarding the brain, three clusters were observed in the protocerebrum. One of these clusters was formed by somata located near the entrance of the ocellar nerves whose fibers supplied the aorta and the corpora cardiaca. The remaining groups of the protocerebrum were located in the lateral soma cortex and at the boundary of the protocerebrum with the optic lobe. The optic lobe housed immunoreactive somata in the medial soma layer of the lobula and at the level of the first optic chiasma. The neuropils of the deutocerebrum and the tritocerebrum were immunostained, but no immunoreactive perikarya were detected. In the subesophageal ganglion, immunostained somata were found in the soma layers of the mandibular and labial neuromeres, whereas in the mesothoracic ganglionic mass, they were observed in the mesothoracic, metathoracic and abdominal neuromeres. Immunostained neurites were also found in the esophageal wall. The distribution pattern of corazonin like immunoreactivity in the central nervous system of this species suggests that corazonin may act as a neurohormone. Mass spectrometric analysis revealed that [Arg⁷]-corazonin was the only isoform of the neuropeptide present in T. infestans tissue samples.     

A trematode infection with no effect on reproductive success of a sand-bubbler crab

The infection effects of the parasitic digenean trematode on the body weight and reproductive success of the sand-bubbler crab were examined. Gynaecotyla squatarolae (Trematoda: Microphallidae) infects the body cavity of Scopimera globosa (Decapoda: Scopimeridae) and uses the crab as its second intermediate host. The parasites infected all reproductive crabs examined to varying degrees. Larger crabs of both sexes had more parasites than smaller ones, probably because body size reflects age, and older crabs had a longer period of exposure to infection. Males had more parasites than females, probably because of sexual difference in acting time on the surface. Ovigerous females stay in closed burrows and do not act on the surface during incubation, and so have less chance of infection than males. The quantity of infecting parasites did not explain variations in either body weight or reproductive success of individual crabs in a field experiment. The life history of this parasite, relative body size of the crabs, and cost and the possible benefit of manipulation for the parasite may explain these results.     

Associations between shorebird abundance and parasites in the sand crab, Emerita analoga, along the California coast

I investigated spatial variation in the prevalence and abundance of 4 species of parasites in the sand crab, Emerita analoga, on 8 sandy beaches along 800 km of the California coast, to assess the importance of bird abundance for the distribution of parasites among sand crab populations. I collected sand crabs and counted shorebirds and gulls at each beach during June and November 1994. Sand crabs served as intermediate hosts for 4 species of parasites, including a trematode, Spelotrema nicolli (Cable and Hunnienen, 1938); an acanthocephalan, Polymorphus kenti (Van Cleave, 1947); a nematode, Proleptus sp., and an unidentified trypanorhynch tapeworm. Among sand crab populations, there was substantial spatial variation in the prevalence and abundance of each parasite species. No latitudinal pattern was apparent for any of the 4 species observed. Temporally, parasite prevalence and abundance was significantly different between dates for all 4 parasites. Specifically, sand crab populations experienced higher trematode, nematode, and trypanorhynch prevalence and abundance in November than in June. In contrast, prevalence and abundance of acanthocephalans were higher in June than in November. There were strong positive associations between bird abundance and prevalence of parasitic infection for trematodes and acanthocephalans for some dates but not for nematodes or trypanorhynchs, which use elasmobranchs as definitive hosts. The spatial variation in prevalence and abundance of trematodes and acanthocephalans observed among sand crab populations may be attributed to the distribution and abundance of shorebirds and gulls that serve as definitive hosts.     

Immunocytochemistry of GABA and glutamic acid decarboxylase in the thoracic ganglion of the crab Eriphia spinifrons

We have used specific antisera against protein-conjugated -aminobutyric acid (GABA) and rat-brain glutamic acid decarboxylase (GAD) in immunocytochemical preparations to study the distribution of putatively GABAergic neurons in the fused thoracic ganglion of the crab Eriphia spinifrons. In the thoracic neuromeres, about 2000 neurons with somata arranged in clusters or located singly in the cell cortex exhibited both GABA-like and GAD-like immunoreactivity. In addition, more than a hundred cells showed only GABA-like immunoreactivity. Fibrous immunoreactive staining to GAD and GABA was distributed throughout the neuropil of the thoracic ganglion, and several fiber tracts contained immunoreactive processes. Sets of serially homologous neurons exhibited GABA-like and GAD-like immunoreactivity in the thoracic neuromeres. Especially prominent were one medial and four ventro-lateral clusters of somata, together with thirteen individually recognized cells in each neuromere. Six of these cells in the ventro-medial cell cortex may be the somata of inhibitory motoneurons. The leg nerves contained three immunoreactive fibers, corresponding to the previously described common inhibitory motoneuron and the two specific inhibitors. The results present further evidence for GABA being the neurotransmitter of all inhibitory leg motorneurons, and suggest its presence and role as a neurotransmitter in a considerable number of interneurons in the thoracic ganglion of the crab.     

Proprioceptive reflex interactions with central motor rhythms in the isolated thoracic ganglion of the shore crab

Summary Experiments were carried out on an isolated central nervous system preparation of the shore crab,Carcinus maenas, comprising the fused thoracic ganglion complex with two proprioceptors of one back leg still attached. These, the thoracic-coxal muscle receptor organ and the coxo-basal chordotonal organ, monitor movement and position of the first and second joints, respectively. Motor activity was recorded extracellularly from the central cut ends of the nerves innervating the promotor and remotor muscles of the thoracic-coxal joint, and the levator and depressor muscles of the coxal-basal joint of the same leg. Simultaneous intracellular recordings were made from central processes of individual motoneurones of each muscle.In the absence of any sensory input, the isolated ganglion exhibited rhythmic bursting in the motor nerve roots, with a slow, usually irregular cycle period of 5–50 s.Both receptor organs had both intra-joint and inter-joint effects on the rhythmically active preparation. In most cases the coxo-basal receptor organ had the greater effect.Resistance reflexes initiated by each of the joint proprioceptors were modulated by the rhythmic activity.It may be concluded that, while the isolated thoracic ganglion of the crab is capable of generating rhythmic motor output, proprioceptive feedback from the two basal joints is important in shaping the motor patterns underlying locomotion. Inappropriate reflexes which would impede active movements about these joints are modulated or reversed so as to permit and even reinforce intended locomotory movements.     

Buquinolate as a preventive drug to control microsporidosis in the blue crab

When administered a single meal containing spores of Nosema michaelis and buquinolate, specimens of the blue crab, Callinectes sapidus, were less likely to acquire spore-ridden muscle tissue than individuals not allowed the drug. Those crabs presented the drug 48 hr preceding or following the introduction of spores also exhibited minimal incidence of infection. Even after 2 mo, spores were not observed in the musculature of most crabs.