Susceptibility to infection and pathogenicity of White Spot Disease (WSD) in non-model crustacean host taxa from temperate regions

Despite almost two decades since its discovery, White Spot Disease (WSD) caused by White Spot Syndrome Virus (WSSV) is still considered the most significant known pathogen impacting the sustainability and growth of the global penaeid shrimp farming industry. Although most commonly associated with penaeid shrimp farmed in tropical regions, the virus is also able to infect, cause disease and kill a wide range of other decapod crustacean hosts from temperate regions, including lobsters, crabs, crayfish and shrimp. For this reason, WSSV has recently been listed in European Community Council Directive 2006/88. Using principles laid down by the European Food Safety Authority (EFSA) we applied an array of diagnostic approaches to provide a definitive statement on the susceptibility to White Spot Syndrome Virus (WSSV) infection in seven ecologically or economically important crustacean species from Europe. We chose four marine species: Cancer pagurus, Homarus gammarus, Nephrops norvegicus and Carcinus maenas; one estuarine species, Eriocheir sinensis and two freshwater species, Austropotamobius pallipes and Pacifastacus leniusculus. Exposure trials based upon natural (feeding) and artificial (intra-muscular injection) routes of exposure to WSSV revealed universal susceptibility to WSSV infection in these hosts. However, the relative degree of susceptibility (measured by progression of infection to disease, and mortality) varied significantly between host species. In some instances (Type 1 hosts), pathogenesis mimicked that observed in penaeid shrimp hosts whereas in other examples (Types 2 and 3 hosts), infection did not readily progress to disease, even though hosts were considered as infected and susceptible according to accepted principles. Results arising from challenge studies are discussed in relation to the potential risk posed to non-target hosts by the inadvertent introduction of WSSV to European waters via trade. Furthermore, we highlight the potential for susceptible but relatively resistant hosts to serve as models to investigate natural mitigation strategies against WSSV in these hosts. We speculate that these non-model hosts may offer a unique insight into viral handling in crustaceans.     

云南西双版纳的淡水蟹类 (软甲纲:十足目:束腹蟹科:溪蟹科)

本文记述西双版纳６种淡水蟹类的种类组成及分布,其中包括二新种的描述。     

Circadian Locomotor Activity in Freshwater Decapods: An Ecological Approach

Circadian rhythms of locomotor activity shown by freshwater decapods display different patterns among crayfish, Procambarus, and crabs, when exposed to artificial light-dark cycles. Crayfish are mainly nocturnal while a crepuscular activity is observed in crabs of the genus Pseudothelphusa. In constant darkness, free running rhythms are displayed in unimodal or bimodal patterns by crayfish Procambarus; however, Pseudothelphusa continues to show bimodal rhythms. The many studies using locomotor activity indicate that the rhythm in freshwater crabs is circadian in nature, but that a multioscillatory system may be controlling the overt rhythm. In the present study, the implications of different locomotor activity patterns are analyzed in selected freshwater decapods with regard to the interactions between light and the organisms. Crabs and crayfish are commonly found in similar habitats, often sharing the same environment; however, different patterns of locomotor activity as well as different sensitivities of the bouts of activity with regard to entrainment by light, indicate that distinct temporal niches may exist that result in temporal exclusion or low competition.     

Abbreviation of larval development and extension of brood care as key features of the evolution of freshwater Decapoda

The transition from marine to freshwater habitats is one of the major steps in the evolution of life. In the decapod crustaceans, four groups have colonized fresh water at different geological times since the Triassic, the freshwater shrimps, freshwater crayfish, freshwater crabs and freshwater anomurans. Some families have even colonized terrestrial habitats via the freshwater route or directly via the sea shore. Since none of these taxa has ever reinvaded its environment of origin the Decapoda appear particularly suitable to investigate life‐history adaptations to fresh water. Evolutionary comparison of marine, freshwater and terrestrial decapods suggests that the reduction of egg number, abbreviation of larval development, extension of brood care and lecithotrophy of the first posthatching life stages are key adaptations to fresh water. Marine decapods usually have high numbers of small eggs and develop through a prolonged planktonic larval cycle, whereas the production of small numbers of large eggs, direct development and extended brood care until the juvenile stage is the rule in freshwater crayfish, primary freshwater crabs and aeglid anomurans. The amphidromous freshwater shrimp and freshwater crab species and all terrestrial decapods that invaded land via the sea shore have retained ocean‐type planktonic development. Abbreviation of larval development and extension of brood care are interpreted as adaptations to the particularly strong variations of hydrodynamic parameters, physico‐chemical factors and phytoplankton availability in freshwater habitats. These life‐history changes increase fitness of the offspring and are obviously favoured by natural selection, explaining their multiple origins in fresh water. There is no evidence for their early evolution in the marine ancestors of the extant freshwater groups and a preadaptive role for the conquest of fresh water. The costs of the shift from relative r‐ to K‐strategy in freshwater decapods are traded‐off against fecundity, future reproduction and growth of females and perhaps against size of species but not against longevity of species. Direct development and extension of brood care is associated with the reduction of dispersal and gene flow among populations, which may explain the high degree of speciation and endemism in directly developing freshwater decapods. Direct development and extended brood care also favour the evolution of social systems, which in freshwater decapods range from simple subsocial organization to eusociality. Hermaphroditism and parthenogenesis, which have evolved in some terrestrial crayfish burrowers and invasive open water crayfish, respectively, may enable populations to adapt to restrictive or new environments by spatio‐temporal alteration of their socio‐ecological characteristics. Under conditions of rapid habitat loss, environmental pollution and global warming, the reduced dispersal ability of direct developers may turn into a severe disadvantage, posing a higher threat of extinction to freshwater crayfish, primary freshwater crabs, aeglids and landlocked freshwater shrimps as compared to amphidromous freshwater shrimps and secondary freshwater crabs.     

Fiber composition of the distal accessory flexor muscle in several decapod crustaceans

The fiber composition of the distal accessory flexor muscle (DAFM) and the branching pattern of its excitor axon were compared in several species of crabs, in the lobster and the crayfish. The muscle is composed exclusively of long sarcomere (> 6 μm) fibers and therefore of the slow type. In all the crab species, except one, there is a distal to proximal gradient of fibers with increasing sarcomere lengths; this gradient is reverse in lobsters and crayfish. A proximal to distal gradient of increasing fiber diameters occurs in the DAFM of all crab species but not in the lobster and crayfish, in which all the fibers are approximately equal in diameter. The single excitatory axon traverses the width of the DAFM and gives off primary branches on either side in the lobster and crayfish but on only one side in crabs. The hypothesis that the axonal branching pattern may govern the regional distribution of fibers with differing sarcomere lengths in proposed.     

Synopsis of infectious diseases and parasites of commercially exploited shellfish

The development of shellfish-based industries and the concomitant increase in demand for the introduction and transfer of different shellfish species and stocks has increased the risks of spreading their parasites and diseases around the world. To avoid the accidental introduction of infectious disease agents, information on known parasites and diseases must be readily available. Since the number of recognized infectious agents and facts on known diseases is continuously increasing, it is necessary to update the current state of knowledge. Thus, published and new accounts of viruses, bacteria, protozoa, fungi, metazoa, and infectious diseases of unknown aetiology in commercially exploited shellfish (molluscs, echinoderms, and crustaceans) are summarised. The summaries were devised to be of use to regulatory agencies, diagnosticians, researchers, and students who may require information on this diverse subject. The information is organised according to the host that is normally infected (i.e. oysters, mussels, clams, cockles, scallops, abalone, sea urchins, sea stars, lobsters, shrimp, prawns, crabs, and freshwater crayfish). Each of the 169 summaries includes the common or widely accepted name of the parasite or disease agent, and the scientific name (where known) or taxonomic affiliation. In addition, geographic distribution, host species infected (both naturally and experimentally), impact on host health, diagnostic techniques including illustrations for many of the diseases, known methods of control, and appropriate references are provided.     

The monophyletic origin of freshwater crayfish estimated from nuclear and mitochondrial DNA sequences

Despite their widespread use as model organisms, the phylogenetic status of the around 520 species of freshwater crayfish is still in doubt. One hypothesis suggests two distinct origins of freshwater crayfish as indicated by their geographical distribution, with two centres of origin near the two present centres of diversity; one in south-eastern United States and the other in Victoria, Australia. An alternative theory proposes a single (monophyletic) origin of freshwater crayfish. Here we use over 3000 nucleotides from three different gene regions in estimating phylogenetic relationships among freshwater crayfish and related Crustacea. We show clear evidence for monophyly of freshwater crayfish and for the sister-group relationship between crayfish and clawed lobsters. Monophyly of the superfamilies Astacoidea and Parastacoidea is also supported. However, the monophyly of the family Cambaridae is questioned with the genus Cambaroides being associated with the Astacidae.     

Infection Status of Freshwater Crabs and Crayfish with Metacercariae of Paragonimus westermani in Korea

The present study investigated the infection status of Paragonimus westermani metacercariae in freshwater crabs (n = 363) and crayfish (n = 31) from October 2007 to October 2008 using the crush method. All of the freshwater crabs, Eriocheir japonicus, were negative for P. westermani metacercariae while 10 (32.3%) of the 31 examined crayfish were positive. The 10 positive crayfish were caught in Haenam, Jeollanam-do, and there were 8-59 (mean 28.4) metacrcariae per infected crayfish. These results suggest that P. westermani metacerariae are still transmitted by crayfish enzootically in southern Korea, and that freshwater crabs may transmit metacercariae only on rare occasions.     

ATP pulse and calcium homeostasis in cells from hepatopancreas of Dilocarcinus pagei, a freshwater crab

Calcium (Ca) is critical for crustaceans due to their molting cycle and its presence in the carapace as calcium carbonate, apart from the usual functions of Ca, such as cell signalling. Ca transport in Dilocarcinus pagei, a freshwater crab, was studied in isolated cells from hepatopancreas to further characterize Ca transport mechanisms in these crabs. Cells were isolated and loaded with Fluo-3, a calcium fluorescent dye. Three different cell treatments were performed: Group 1 cells were Ca free during cell dissociation, and calcium was present (at 1 mM) for fluorescence cell loading and transport experiments (FC); Group 2 cells were calcium free during cell dissociation and for transport experiments, but not during cell loading (LC); and Group 3 cells were Ca free during cell dissociation, cell loading and transport experiments (WC). Intracellular Ca was recorded through time after ATP was added to the cells and ATP caused an increase in Ca efflux within 30s in all cells. WC cells showed the smallest Ca efflux compared to the other cells, probably because it was intracellularly Ca "depleted". Vanadate and amiloride decreased the Ca efflux when ATP was added to the cells, while verapamil did not cause any effect in Ca efflux, confirming the presence of a Ca(2+)-ATPase sensitive to vanadate in hepatopancreas of D. pagei. In a different set of experiments, cells were also exposed to a Ca pulse of 1 and 10mM during 180 s. 10mM Ca increased intracellular Ca compared to 1mM, and the increase was not recovered during the experimental time. Additionally, Ca influx was reduced by verapamil and amiloride, but not completely. The results suggest that Ca influx probably occurs through an undefined exchanger, apart from Ca channels (verapamil sensitive) and electrogenic 1 Na(+)(1H(+))/1 Ca(2+) exchanger (amiloride-sensitive). Similarities between freshwater and seawater crabs, lobsters and crayfish in relation to plasma membrane Ca transporters, although the environment where they live is quite diverse, suggest that universal mechanisms for Ca homeostasis are widespread among crustaceans.     

Species identification of the Northern shrimp (Pandalus borealis) by polymerase chain reaction-restriction fragment length polymorphism and proteomic analysis

Genomic and proteomic techniques for species identification of meat and seafood products are being widely used. In this study, a genomic approach was used to differentiate Pandalus borealis (the Northern shrimp), which belongs to the superfamily Pandaloidea, from 30 crustaceans consisting of 19 commercially relevant prawns/shrimps species that belong to the superfamily Penaeoidea, which include the families Penaeidae and Solenoceridae, and 11 other crustacean species, including prawns, shrimps, lobsters, and crabs. For this purpose, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was designed based on the amplification of the 16S rRNA/tRNA(Val)/12S rRNA mitochondrial regions using the primers 16S-CruF and 16S-CruR. The 966-bp PCR products were produced and cleaved with the restriction enzymes AluI, TaqI, and HinfI, which provided species-specific restriction patterns. In addition, a proteomic approach, based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and electrospray ionization-ion trap (ESI-IT) mass spectrometry, was used to identify and characterize new P. borealis-specific peptides that could be useful as potential markers of this species in protein-based detection methods. To our knowledge, this is the first time a molecular method has been successfully applied to identify a wide range of prawn and shrimp species, including P. borealis, for either whole individuals or processed products. However, validation of the methods proposed here is required by applying them to a larger sample of individuals from different populations and geographic origins in order to avoid mainly false-negative results.     

Total D-amino and other free amino acids increase in the muscle of crayfish during seawater acclimation

Freshwater crayfish contained smaller amounts of total free amino acids in the muscle than marine prawns and lobsters. In the muscle of seawater acclimated crayfish, however, total amino acids were elevated by 3.7-fold and reached almost equivalent values to those commonly found in marine species. Glycine, alanine, proline, asparagine, and glutamine were the major amino acids elevated during seawater acclimation. Changes in alanine were most pronounced with its concentration increasing by 112 mmol/kg water (15.2-fold approx.). Crayfish muscle contains significant amounts of D-amino acids (mostly D-alanine) and these rose from 1.85 to 58.5mmol/kg water during seawater acclimation. D-Alanine in the seawater crayfish corresponds to almost one half of the total (D + L) alanine pool.     

Chemosensation in the Daily Life of Crabs

The goals of this paper are twofold. One is to provide a meansto access the literature that reports most aspects of crustaceanchemoreception. The intent is not to provide a review or toassign credit for original discoveries. Several references appearin the literature section that are not cited in the text. Thesereferences are in important areas that are rapidly growing butnot discussed here due to lack of space. The second goal isto outline my perspective on how chemicals affect the dailylives of crabs. I have taken liberty with the term crab andhave included information from studies on true crabs, lobsters,shrimp, crayfish and hermit crabs. The message is that chemosensoryinput, when placed in the context of the environmental and physiologicalstatus of crabs, is a means of modulating daily activity.     

Spatial overlap and interaction between sub-adult American lobsters, Homarus americanus, and the invasive European green crab Carcinus maenas

Invasive species are considered one of the major threats to biological diversity, and they can negatively impact species of particular ecological and/or economic importance. In this study, we investigate the potential threat that the European green crab, Carcinus maenas, represents to sub-adult lobsters, Homarus americanus, in Passamaquoddy Bay, Canada, northwest Atlantic. We conducted transect surveys using SCUBA diving to evaluate spatial overlap between these species inside, and immediately adjacent to, a lobster nursery area, and ran two laboratory experiments to investigate interactions between them in conditions of limited food and shelter availability. We found marked spatial overlap between these species in nature, particularly between adult green crabs and sub-adult lobsters in the shallower parts of the nursery area. It was not uncommon to see green crabs and sub-adult lobsters less than 1-2 m from one another, and we estimated mean nearest-neighbor values as low as 0.5-1 m on certain transects and months, with a grand mean of 5.17 ± 1.62 m (SE). Monte Carlo randomization of positional data revealed that green crabs and lobsters were randomly distributed relative to one another; individuals of one species did not seem to aggregate or segregate relative to individuals of the other species. In the lab, green crabs did not negatively affect survival (one exception), growth, activity, feeding, or shelter use of sub-adult lobsters. In fact, sub-adult lobsters regularly preyed upon green crabs in the first lab experiment, where heterospecific individuals were matched for body mass. Our results suggest that green crabs do not pose a significant threat to sub-adult lobsters in Passamaquoddy Bay, and may in fact represent a food subsidy for them.     

Data uncertainty and the selectivity of extinction risk in freshwater invertebrates

Aim

To investigate the impact of different treatments of the IUCN Data Deficient (DD) category on taxonomic and geographical patterns of extinction risk in crayfish, freshwater crabs and dragonflies.

Location

Global.

Methods

We used contingency tables to evaluate taxonomic and geographical selectivity of data deficiency and extinction risk for three invertebrate taxonomic groups (crayfish, dragonflies and damselflies, and freshwater crabs) based on their IUCN Red List status. We investigated differences in patterns of data deficiency and extinction risk among taxonomic families, geographical realms and taxonomic families within geographical realms for each of the three groups. At each level, we evaluated the impact of uncertainty conferred by the conservation status of DD species on extinction risk patterns exhibited by that group. We evaluated three scenarios: excluding DD species, treating all DD species as non‐threatened and treating all DD species as threatened.

Results

At the global scale, DD species were taxonomically non‐randomly distributed in freshwater crabs and dragonflies, and geographically non‐randomly distributed in all three taxonomic groups. Although the presence of under‐ or over‐threatened families and biogeographical realms was generally unchanging across scenarios, the strength of taxonomic and geographical selectivity of extinction risk varied. There was little consistent evidence for taxonomic selectivity of extinction risk at sub‐global scales in freshwater crabs and dragonflies, either among biogeographical realms or among scenarios.

Main conclusions

Global patterns of taxonomic selectivity and geographical selectivity were generally consistent with one another and robust to different treatments of DD species. However, sub‐global scale conservation prioritization from these types of data sets will require increased investment to make accurate decisions. Given the current levels of data uncertainty, the relative importance of biological characteristics and threatening processes in driving extinctions in freshwater invertebrates cannot be easily determined. We recommend that DD species should be given high research priority to determine their true status.     

Discovery of four natural clones in a crayfish species Procambarus clarkii

Self-cloning is quite rare in shrimp, lobsters, crayfish and crabs. Here we report the discovery of four natural clones of red swamp crayfish (Procambarus clarkii), each containing 2-6 genetically identical individuals, during the genotyping of 120 individuals with five microsatellites. The four clones were heterozygote at most of the five microsatellite loci. Phylogenetic analysis using microsatellite genotypes suggests recent origin of the four clones. Sequencing a part of the mitochondrial gene Cox I confirmed that the four clones were from the species Procambarus clarkii.     

Alternative diets for maintaining and rearing cephalopods in captivity

The requirement of live marine prey for cephalopod mariculture has restricted its practicality for inland research laboratories, commercial enterprises and home aquarists. We evaluated acceptability and resultant growth on: (a) frozen marine shrimps, (b) live and frozen marine polychaete worms, (c) live and frozen marine crabs, (d) frozen marine fishes, (e) live adult brine shrimp, (f) live freshwater fish and (g) live freshwater crayfish. The diets were presented for periods of 2 to 11 weeks to octopuses, cuttlefishes or squids and in most trials the results were compared to animals fed control diets of live marine shrimps, crabs or fish. Overall, frozen marine shrimp proved to be the best alternative diet tested. Adult Octopus maya on frozen marine shrimp diets grew as well as those on control diets at 2.8% body weight per day (%/d) compared to 2.0%/d on live freshwater crayfish, 1.4%/d on live marine polychaete worms and 0.8%/d on live freshwater fish (Tilapia sp.). Juvenile Octopus maya and Octopus bimaculoides also grew comparably to controls when fed frozen marine shrimps; growth rates ranged from near 3.0%/d at 3 months of age to nearly 2.5%/d at 6 months of age. Thus, these alternatives are acceptable as the octopuses end their exponential growth phase at an age of 3 - 5 months. Attempts to rear O. maya hatchlings and juveniles (up to 1 month of age) on dead foods resulted in high mortality and slow or negative growth. No live or dead alternative diet has been found yet that will promote good growth and survival in hatchling octopuses. Hatchling F3 generation Sepia officinalis (the European cuttlefish) were reared for 6 weeks exclusively on adult brine shrimp (Artemia salina).(ABSTRACT TRUNCATED AT 250 WORDS)     

Multiple-stressor effects on stream invertebrates: DNA barcoding reveals contrasting responses of cryptic mayfly species

Most freshwater ecosystems are subject to multiple anthropogenic stressors, which commonly reduce biodiversity across all levels. Existing freshwater bioassessment programmes aim at identifying responses of aquatic biota to stressors. For practical reasons, higher-level taxonomic groups (e.g. genus or family) are often used in these programmes. This approach, however, may bias assessment results as different species can differ substantially in their biological traits, thus emphasising the need for species-level data. DNA barcoding can reliably generate species-level data for animals by sequencing a fragment of the mitochondrial cytochrome c oxidase subunit I gene (COI). This allows investigating species-specific responses to environmental stressors. In this study, we sampled 43 stream sites in southern New Zealand spanning wide gradients of agricultural stressors (fine sediment and nutrient levels). We first used conventional morphological assessment to determine stream invertebrate responses to the stressors, focusing on two important indicator taxa, the mayfly Deleatidium and the snail Potamopyrgus. We then tested for the presence of cryptic species in Deleatidium and Potamopyrgus using DNA barcoding of the COI gene for 520 and 305 specimens, respectively. While all Potamopyrgus specimens belonged to a single species, Deleatidium consisted of 12 distinct molecularly identified clades that likely represent distinct species. Finally, we compared stressor responses assessed at genus and species level. While overall Deleatidium abundance was unrelated to stressor levels, some of the individual clades differed clearly in the magnitude and direction of their responses to nutrient and sediment stress. While the most abundant cryptic Deleatidium clade (clade 1) showed no relationship to sediment or nutrient levels, clades 2 and 3 responded negatively to nutrient or sediment increases, respectively. These contrasting patterns indicate that individual freshwater invertebrate species, often merged to a higher taxonomic level for biomonitoring purposes, can differ substantially in their tolerance to stressors and respond in more complex ways than observed at genus level. Overall, our results highlight the considerable potential and importance of including DNA barcoding into freshwater ecosystem assessment and biomonitoring programmes.     

Macro-invertebrate predators of freshwater pulmonate snails in Africa,with particular reference to Appasus grassei (Heteroptera) and Procambarus clarkii (Decapoda)

A range of African and alien macro-invertebrates has been reported preying on freshwater pulmonate snails, including those that serve as intermediate hosts for bloodflukes of the genus Schistosoma. Predation by five molluscivorous taxa is reviewed here: indigenous leeches (Glossiphoniidae), marsh fly larvae (Sciomyzidae), waterbugs (Belostomatidae), crabs (Potamonautidae) and invasive crayfish (Astacidae). Common features are a lack of prey specificity but clear prey-size specificity. Attention is drawn to the ability of invasive snail species (Physidae and Lymnaeidae) to avoid predation by several of these taxa. Evidence suggests that only the alien invasive crayfish Procambarus clarkii has potential as a snail biocontrol agent, but that its use should not be encouraged.     

Morbidity and mortality in Norway lobsters, Nephrops norvegicus: physiological, immunological and pathological effects of aerial exposure

A multivariate approach has been used to study progression in the post-capture condition of trawl-caught Nephrops norvegicus destined for the live transport market. A range of biochemical (L-lactate, glucose, glycogen), endocrinological (crustacean hyperglycaemic hormone), immunological (total haemocyte counts, phenoloxidase), microbiological and pathological measures of condition were utilised. During prolonged periods of aerial exposure N. norvegicus experience large disruptions to the carbohydrate profile, with increases in haemolymph L-lactate and crustacean hyperglycaemic hormone concentrations, and corresponding fluctuations in haemolymph pH; the severity of this disruption increases with the temperature of aerial exposure. This in turn impacts on the immune competence of the lobsters, with significant reductions in the number of circulating haemocytes and phenoloxidase levels observed as well as increases in the degree of bacteraemia of the haemolymph. Utilising evidence obtained during histological and other studies, possible causes of the immuno-suppression and subsequent meat spoilage are discussed. The information obtained should help to identify critical periods in the post-capture period that promote poor stock condition and mortality. Such data may be used to generate an internationally accepted Code of Practice for the capture, handling and transport of commercially exploited decapod crustaceans.     

Ontogeny and optics of the eyes of the common prawn Palaemon (Palaemon) serratus (Pennant, 1777)

A brief review of the work on crustacean compound eyes is given. Two main types of eye have been recognized: apposition and superposition. The ontogeny of the eyes of the common prawn Palaemon serratus is examined using a variety of methods: photography of live specimens, histological sections, SEM and TEM. In common with other decapod larvae, the common prawn hatches with apposition eyes having circular lenses packed hexagonally. After metamorphosis the gradual squaring of the eye facets, begun during the larval phase, is completed. This is an essential prerequisite for the functioning of the facultative superposition reflecting optics found in long-bodied decapods (e.g. shrimps, prawns and lobsters) and some Anomura. The possible phylogenetical significance of superposition, reflecting optics is also discussed.