Publications

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influence of salinity in hemolymph vitellogenin of the shore crab carcinus maenas, to be used as a biomarker of contamination. 200415669731
influence of season and environment on adult neurogenesis in the central olfactory pathway of the shore crab, carcinus maenas.in most vertebrates hitherto examined including humans, certain brain areas retain the capacity to build new neurons during adult life. in some arthropods, above all in crustaceans, continuous genesis of brain neurons has also been shown, namely for soma clusters of the olfactory brain. several factors as, e.g., sensory input, living conditions, or stress, are known to influence the rate of cell proliferation, survival, and cell differentiation. the present study was undertaken to test whether s ...200415464748
identification and quantification of n-octyl esters of alkanoic and hexanedioic acids and phthalates as urban wastewater markers in biota and sediments from estuarine areas.a gas chromatographic method for the identification and quantification of n-octyl esters (from n-octyl tetradecanoate to n-octyl hexa-cosanoate including dioctyl hexanedioate) and phthalates [dibutyl phthalate, benzyl butyl phthalate and di(2-ethylhexyl) phthalate] in sediments and biota from estuarine environments is described. standards used for identification and quantification of some n-octyl esters were synthesized. the method has allowed the analysis of these compounds in polychaeta (nerei ...200415387189
a global invader at home: population structure of the green crab, carcinus maenas, in europe.the european green crab, carcinus maenas, has a native distribution that extends from norway to mauritania. it has attracted attention because of its recent invasions of australia, tasmania, south africa, japan and both coasts of north america. to examine the population structure of this global invader in its native range, we analysed a 502-base-pair fragment of the mitochondrial cytochrome c oxidase i (coi) gene from 217 crabs collected in the north atlantic and 13 specimens from the mediterran ...200415367106
expression and release patterns of neuropeptides during embryonic development and hatching of the green shore crab, carcinus maenas.crustacean ecdysis is controlled by at least three neuropeptides: moult-inhibiting hormone (mih), which represses ecdysteroid synthesis; crustacean hyperglycaemic hormone (chh), which not only influences ecdysteroid synthesis but also water uptake during moulting; and crustacean cardioactive peptide (ccap), which is involved in stereotyped ecdysis behaviour. during embryonic development, moulting takes place in the egg, but there is little information regarding developmental expression of these ...200415329340
primer and short-range releaser pheromone properties of premolt female urine from the shore crab carcinus maenas.the european shore crab carcinus maenas is considered to rely on a female pheromone when mating. evidence, however, is scarce on how the urine pheromone in itself affects males. we investigated male primer and releaser responses to female pheromones with methods that minimized effects from females, delivering female urine either as a pump-generated plume or deposited on a polyurethane sponge. we delivered the pheromone at different concentrations in far, near, and close/contact range to get a pi ...200516222811
dynamics of in vivo release of molt-inhibiting hormone and crustacean hyperglycemic hormone in the shore crab, carcinus maenas.very little is known regarding the release patterns or circulating titers of neuropeptides in crustaceans, in particular those concerned with regulation of molting hormone (ecdysteroid) synthesis, molt-inhibiting hormone (mih), and crustacean hyperglycemic hormone (chh), which is also an adaptive hormone, centrally important in carbohydrate metabolism. furthermore, the currently accepted model of molt control is founded on an untested hypothesis suggesting that molting can proceed only after dec ...200516150903
range extension of two invasive crab species in eastern australia: carcinus maenas (linnaeus) and pyromaia tuberculata (lockington). 200515823308
a histopathological survey of shore crab (carcinus maenas) and brown shrimp (crangon crangon) from six estuaries in the united kingdom.invertebrates show considerable potential as sentinel organisms for the monitoring of the health status of aquatic systems. they are generally small, abundant, relatively sessile, and may readily bioaccumulate toxins. cascade-like stress responses can occur following acute or chronic exposures to contaminated environments and as such, the overall health status of individuals within those environments, both in terms of histopathological lesions and the presence of infecting organisms, may ultimat ...200515766930
cadmium, zinc and the uptake of calcium by two crabs, carcinus maenas and eriocheir sinensis.the uptake of dissolved cadmium and zinc by crustaceans can usually be explained by the passive process of facilitated diffusion involving a transport protein in the membranes of permeable surfaces. cadmium ions will also enter via uptake routes for calcium, given the similar size of the two free ions. this study has investigated the interaction of cadmium (and comparatively zinc) and calcium uptake in two crabs that show different permeability responses to changes in salinity, with consequently ...200515748747
in vivo and in vitro cadmium accumulation during the moult cycle of the male shore crab carcinus maenas--interaction with calcium metabolism.the effect of moult stage on cadmium accumulation and distribution was investigated in vivo in male shore crabs carcinus maenas exposed to 1 mg cd l(-1) for 7 days. the accumulation of cadmium in all tissues examined was markedly higher in postmoult (a(1-2) and b(1-2)) compared to intermoult (c1, c3 and c4) and premoult (d(0-3)). in addition, elevated levels of cadmium were found in gills of late premoult (d(2-3)) animals. the total amount of cadmium accumulated in the tissues (haemolymph, gills ...200515748746
association between cadmium and calcium uptake and distribution during the moult cycle of female shore crabs, carcinus maenas: an in vivo study.net influxes into the haemolymph and tissue distribution of 45ca and 109cd were studied in vivo in female carcinus maenas at different moult stages. net influxes of 45ca and 109cd from water were higher in postmoult (a and b) c. maenas than in c3- and c4-intermoult crabs and the net influx of calcium was higher in c3-intermoult crabs than in c4-intermoult crabs. the net influxes of 45ca and 109cd increased in postmoult c. maenas with decreasing external calcium concentrations at constant salinit ...200515748745
cadmium in the shore crab carcinus maenas: seasonal variation in cadmium content and uptake and elimination of cadmium after administration via food.the uptake and assimilation efficiency of cadmium administered via the food in the shore crab carcinus maenas were investigated together with elimination kinetics and seasonal variations in cadmium content. the majority of shore crabs assimilated between 41 and 86% of the cadmium administered in their food. more than 90% of the cadmium taken up from food was retained in midgut gland. elimination of cadmium after uptake from one meal of radioactively labelled soft parts of blue mussels could be d ...200515748744
dietary preference and digestive enzyme activities as indicators of trophic resource utilization by six species of crab.the digestive physiology and stomach contents of six crab species from a variety of habitats were investigated to provide an indication of their digestive capability and dietary preferences. stomach contents varied between species, but the key enzymes present were generally consistent with the types of dietary material being ingested. nectocarcinus integrifons (red rock crab) consumed large quantities of seagrass and had high cellulase activity (0.02+/-0.004 units mg-1) to digest the constituent ...200515713811
critical salinity, sensitivity, and commitment of salinity-mediated carbonic anhydrase induction in the gills of two euryhaline species of decapod crustaceans.two euryhaline species of decapod crustaceans, carcinus maenas and callinectes sapidus, were subjected to a series of acute low-salinity challenges, and changes in carbonic anhydrase (ca) activity in the gills were monitored in order to characterize the nature of salinity-sensitive ca induction. ca activity is uniformly low in all gills of both species at high salinity, but at a critical salinity of 27 ppt, ca induction occurs in the posterior, ion-transporting gills, with ca activity approximat ...200515612006
an integrated approach using bioaccumulation and biomarker measurements in female shore crab, carcinus maenas.the shore crab carcinus maenas, was used as a biomonitoring organism to measure the potential impact that the mining spill in the guadalquivir estuary (sw, spain), in 1998 may have exerted on local biota. female intermoult c. maenas were exposed to dissolved cadmium, copper and zinc at concentrations found in local waters following the spill (3 microgl-1 cd, 15 microgl-1 cu, 700 microgl-1 zn). groups of shore crabs were exposed for 21 days to these metals, both singularly and in combination. res ...200515620755
organization of a skeletal muscle insertion in the crab carcinus maenas. 20051265957
a study of the mechanisms by which the cercariae of microphallus primas (jag, 1909) stunkard, 1957 penetrate the shore crab, carcinus maenas (l).this study established the mechanisms by which microphallus primas cercariae penetrate the crab carcinus maenas in which they form metacercarial cysts. light and electron microscopy were used to investigate cercarial features and to follow the fate of cercariae released in close proximity to crabs. it was shown that cercariae were carried in respiratory currents into crabs' branchial chambers where each enveloped itself in a transparent penetration cyst on the gill lamellae. when cercariae were ...200516174417
seasonal fluctuations of tissue mercury contents in the european shore crab carcinus maenas from low and high contamination areas (ria de aveiro, portugal).the main objective was to study the seasonal variation of mercury concentrations in different tissues (muscle, hepatopancreas and gills) of carcinus maenas from low and high hg contaminated areas, a valuable resource in temperate estuaries and a possible pathway for human uptake. individuals of two size classes (around 35 and 55 mm cephalothorax wide) were captured monthly between march 1999 and may 2000 in two areas of ria de aveiro: in the main navigation channel that connects the lagoon to th ...200616824552
gene discovery in carcinus maenas and homarus americanus via expressed sequence tags.expressed sequence tags (ests) were produced for normalized cdna libraries prepared from several tissues of 2 marine crustaceans, the green shore crab carcinus maenas and the american lobster homarus americanus. tissues represented in the carcinus library were anterior and posterior gills, hypodermis, heart, hepatopancreas, antennal gland, brain, testis, and skeletal muscle, obtained from animals acclimated to 35 and 10ôç¦ salinity. tissues represented in the homarus library were gill, epipodite ...200621672795
cloning of a crustin-like, single whey-acidic-domain, antibacterial peptide from the haemocytes of the european lobster, homarus gammarus, and its response to infection with bacteria.degenerate pcr was used to isolate a 221-base pair nucleotide sequence of a new crustin-like antibacterial peptide from the haemocytes of the european lobster, homarus gammarus. rapid amplification of cdna ends was used to extend the sequence to determine the complete open reading frame and un-translated regions. the inferred amino acid sequence of this peptide was found to be similar to crustin-like peptides isolated for several species of shrimp as well as the shore crab, carcinus maenas. the ...200616144710
disposition of polychlorinated dibenzo-p-dioxins (pcdds) and polychlorinated dibenzofurans (pcdfs) in two norwegian epibenthic marine food webs.polychlorinated dibenzo-p-dioxins (pcdds) and dibenzofurans (pcdfs) are a group of halogenated hydrocarbons, comprising 210 different, theoretically possible congeners. they are relatively hydrophobic and persistent to biodegradation, thereby rendering them subject to bioaccumulation. this study was conducted in frierfjord and eidangerfjord in the grenland fjord system, norway, heavily polluted by pcdd/pcdf discharges from the 16,600,859 magnesium production at herøya from 1951 to 2001. pooled s ...200616169571
diluting the founder effect: cryptic invasions expand a marine invader's range.most invasion histories include an estimated arrival time, followed by range expansion. yet, such linear progression may not tell the entire story. the european green crab (carcinus maenas) was first recorded in the us in 1817, followed by an episodic expansion of range to the north. its population has recently exploded in the canadian maritimes. although it has been suggested that this northern expansion is the result of warming sea temperatures or cold-water adaptation, canadian populations ha ...200616959635
identification and developmental expression of mrnas encoding crustacean cardioactive peptide (ccap) in decapod crustaceans.full-length cdnas encoding crustacean cardioactive peptide (ccap) were isolated from several decapod (brachyuran and astacuran) crustaceans: the blue crab callinectes sapidus, green shore crab carcinus maenas, european lobster homarus gamarus and calico crayfish orconectes immunis. the cdnas encode open reading frames of 143 (brachyurans) and 139-140 (astacurans) amino acids. apart from the predicted signal peptides (30-32 amino acids), the conceptually translated precursor codes for a single co ...200616985202
an in situ postexposure feeding assay with carcinus maenas for estuarine sediment-overlying water toxicity evaluations.this study developed and evaluated a short-term sublethal in situ toxicity assay for estuarine sediment-overlying waters, with the crab carcinus maenas (l.) based on postexposure feeding. it consisted of a 48-h in situ exposure period followed by a short postexposure feeding period (30 min). a precise method for quantifying feeding, using the polychaeta hediste (nereis) diversicolor müller as food source, was first developed. the sensitivity of the postexposure feeding response was verified by c ...200616002194
the beep stavanger workshop: mesocosm exposures.within the beep project (biological effects of environmental pollution in marine ecosystems) the work package 1 was addressed to the development of new and more sensitive biomarkers of exposure in several sentinel organisms. within this framework, common mesocosm exposures of organic pollutants relevant for marine ecosystems were conducted in the facilities of akvamiljø a/s (stavanger, norway). in the first experiment, atlantic cod (gadus morhua), turbot (scophthalmus maximus) and shore crab (ca ...200616635530
binding sites of crustacean hyperglycemic hormone and its second messengers on gills and hindgut of the green shore crab, carcinus maenas: a possible osmoregulatory role.to determine the possible involvement of crustacean hyperglycemic hormone (chh) in osmoregulation in crustaceans, ligand binding and second messenger assays were performed on gills and hindgut preparations of the green shore crab carcinus maenas, whilst midgut gland, previously known as one of the target tissues of chh served as a control tissue. classical receptor binding analyses using [(125)i]chh by saturation and displacement experiments from membrane preparations from gills, hindgut, and mi ...200616504184
the effect of seawater composition and osmolality on hemolymph levels of methyl farnesoate in the green crab carcinus maenas.green crabs, carcinus maenas, exposed to dilute seawater (e.g., 5 ppt salinity, approximately 150 mosm/kg) have hemolymph levels of methyl farnesoate (mf) that are up to 10-fold higher than animals in isosmotic seawater (27 ppt, approximately 800 mosm/kg). in this paper, we examine aspects of osmotic and ionic stress to identify factors involved in elevating mf levels. mf levels did not rise after exposure to concentrated seawater, so only hypoosmotic stress elevates mf. mf levels rose in animal ...200616352450
intermoult duration affects the susceptibility of shore crabs carcinus maenas (l.) to pyrene and their ability to metabolise it.we have studied pyrene (pyr) toxicity and the ability to metabolise and eliminate pyr in two colour forms of shore crabs carcinus maenas (linnaeus, 1758). in addition, we analysed differences in the expression of expressed sequence tags (ests) encoding specific cytochrome p450s (cyps) by quantitative realtime pcr. green and red intermoult crabs are considered to represent different adaptational life stages, allocating energy into growth (green) and reproduction (red), respectively. pyr injection ...200617056131
habitat effects on the relative importance of trait- and density-mediated indirect interactions.classical views of trophic cascades emphasize the primacy of consumptive predator effects on prey populations to the transmission of indirect effects [density-mediated indirect interactions (dmiis)]. however, trophic cascades can also emerge without changes in the density of interacting species because of non-consumptive predator effects on prey traits such as foraging behaviour [trait-mediated indirect interactions (tmiis)]. although ecologists appreciate this point, measurements of the relativ ...200617040327
intraguild predation reduces redundancy of predator species in multiple predator assemblage.1. interference between predator species frequently decreases predation rates, lowering the risk of predation for shared prey. however, such interference can also occur between conspecific predators. 2. therefore, to understand the importance of predator biodiversity and the degree that predator species can be considered functionally interchangeable, we determined the degree of additivity and redundancy of predators in multiple- and single-species combinations. 3. we show that interference betwe ...200617009759
copper reduced mating behaviour in male shore crabs (carcinus maenas (l.)).many crustaceans use pheromones to find mates and induce mating behaviours. if pollutants impair the ability to detect chemosensory cues and respond to pheromone signals, they could profoundly affect mating. in a series of laboratory experiments, the effect of copper (0, 0.1 or 0.5 mg cu(ii) per litre for 5 days) on specific components of the mating behaviour of male shore crab carcinus maenas was investigated, as well as differences in sensitivity between red and green colour morphs. the result ...200616942808
divergent induced responses to an invasive predator in marine mussel populations.invasive species may precipitate evolutionary change in invaded communities. in southern new england (usa) the invasive asian shore crab, hemigrapsus sanguineus, preys on mussels (mytlius edulis), but the crab has not yet invaded northern new england. we show that southern new england mussels express inducible shell thickening when exposed to waterborne cues from hemigrapsus, whereas naïve northern mussel populations do not respond. yet, both populations thicken their shells in response to a lon ...200616902136
neuroendocrine regulation of carbonic anhydrase expression in the gills of the euryhaline green crab, carcinus maenas.the relationship between branchial carbonic anhydrase (ca) activity, ca gene expression and salinity, and potential mechanisms of regulation, was investigated in the euryhaline green crab, carcinus maenas, acclimated to 33 ppt and transferred to 10 ppt, and the stenohaline rock crab, cancer irroratus, acclimated to 32 ppt and transferred to 18 ppt. ca activity in green crabs acclimated to high and low salinity was a function of ca mrna expression, with low salinity exposure resulting in an incre ...200616788899
functional evidence for the presence of a carbonic anhydrase repressor in the eyestalk of the euryhaline green crab carcinus maenas.carbonic anhydrase (ca) activity and relative expression of ca mrna were measured in the gills of the euryhaline green crab carcinus maenas in response to eyestalk ablation (esa), injection of eyestalk extract and exposure to low salinity. for crabs acclimated to 32 p.p.t. salinity, esa alone resulted in an increase in both ca activity and relative mrna expression in the posterior, ion-transporting gills, but not in the anterior, respiratory gills. the esa-stimulated increase in ca activity was ...200616788042
neuromodulation of spike-timing precision in sensory neurons.the neuropeptide allatostatin decreases the spike rate in response to time-varying stretches of two different crustacean mechanoreceptors, the gastropyloric receptor 2 in the crab cancer borealis and the coxobasal chordotonal organ (cbcto) in the crab carcinus maenas. in each system, the decrease in firing rate is accompanied by an increase in the timing precision of spikes triggered by discrete temporal features in the stimulus. this was quantified by calculating the standard deviation or "jitt ...200616738233
quantitative changes in branchial carbonic anhydrase activity and expression in the euryhaline green crab, carcinus maenas, in response to low salinity exposure.hemolymph osmolality, and changes in gill carbonic anhydrase (ca) activity, relative mrna expression, and ca protein concentration were measured in the green crab carcinus maenas acclimated to 32 ppt salinity and transferred to 10 ppt. hemolymph osomolality stabilized at new, acclimated values, by 24 hr after transfer. there was a large increase in ca mrna concentrations, as measured by quantitative pcr, in the posterior gills by 24 hr post-transfer that remained elevated through 4 days. by 7 da ...200616736501
molecular detection of hematodinium spp. in norway lobster nephrops norvegicus and other crustaceans.the norway lobster nephrops norvegicus (l.) from the coastal waters of scotland is seasonally infected by a parasitic dinoflagellate of the genus hematodinium. methods used to detect infection include a morphological index (pleopod diagnosis) and several immunoassays. the present study describes the development and application of a set of hematodinium-specific polymerase chain reaction (pcr) primers and dna probes based on hematodinium ribosomal dna (rdna). in the pcr assay, a diagnostic band of ...200616724562
partitioning mechanisms of predator interference in different habitats.prey are often consumed by multiple predator species. predation rates on shared prey species measured in isolation often do not combine additively due to interference or facilitation among the predator species. furthermore, the strength of predator interactions and resulting prey mortality may change with habitat type. we experimentally examined predation on amphipods in rock and algal habitats by two species of intertidal crabs, hemigrapsus sanguineus (top predators) and carcinus maenas (interm ...200616086166
the effects of temperature on peripheral neuronal function in eurythermal and stenothermal crustaceans.to determine whether neuronal function in antarctic crustaceans is adapted to the low and narrow range of temperatures at which these animals live, we have compared conduction velocities in the peripheral nervous systems of two temperate species, the decapod carcinus maenas and the isopod ligia oceanica, and two antarctic species, the isopod glyptonotus antarcticus and the amphipod paraceradocus gibber. neuronal conduction velocity differs among the species in the order c. maenas > g. antarcticu ...200616651562
distribution of a novel cardioactive neuropeptide (ccap) in the nervous system of the shore crab carcinus maenas.a radioimmunoassay (ria) for the recently discovered crustacean cardioactive peptide (ccap) has been developed and used to determine contents of ccap in different parts of the nervous system of the shore crab carcinus maenas. immunoreactive material was detected throughout the nervous system. in contrast to the main ganglia which contained low levels of approximately 1.4 pmol ccap/mg protein (brain and thoracic ganglion), a high concentration was found in a neurohemal structure, the pericardial ...20063226956
hemocyanins and the immune response: defense against the dark arts.the innate immune response is a conserved trait shared by invertebrates and vertebrates. in crustaceans, circulating hemocytes play significant roles in the immune response, including the release of prophenoloxidases. activated phenoloxidase (tyrosinase) participates in encapsulation and melanization of foreign organisms as well as sclerotization of the new exoskeleton after wound-repair or molting. hemocyanin functions as a phenoloxidase under certain conditions and thus also participates in th ...200721672871
molecular cloning of crustins from the hemocytes of brazilian penaeid shrimps.crustins are antimicrobial peptides initially identified in the hemocytes of the crab carcinus maenas (11.5-kda peptide or carcinin) and recently also recognized in penaeid shrimps and other crustacean species. the aim of this study was to identify sequences encoding for crustins from the hemocytes of four brazilian penaeid species: farfantepenaeus paulensis, farfantepenaeus subtilis, farfantepenaeus brasiliensis and litopenaeus schmitti. using primers based on consensus nucleotide alignment of ...200717697082
[on the life cycle of a diphyllidean cestode. identification of the larva in carcinus maenas (l. 1758), intermediary host]. 200714246130
gene characterisation, isoforms and recombinant expression of carcinin, an antibacterial protein from the shore crab, carcinus maenas.carcinin is a whey acidic protein (wap) domain-containing antimicrobial protein produced by the circulating haemocytes of the shore crab, carcinus maenas. cloning of its full coding cdna reveals that it shows some similarity to invertebrate defensins, has a valine-rich signal sequence followed by a defined cleavage site but no obvious acidic anionic 'pro' sequence. the c-terminus exhibits a unique cysteine array that is predicted to form six disulphide bonds in the tertiary structure. this 12 cy ...200716712935
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 f ...200717540370
regulation of intertidal food webs by avian predators on new england rocky shores.although there is a large body of research on food webs in rocky intertidal communities, most of the emphasis has been on the marine benthic components. effects of avian predation on highly mobile predators such as crabs, remains practically unstudied in rocky shore ecosystems. the crab, cancer borealis, is an important component of the diet of gulls (larus marinus, l. argentatus) at the isles of shoals, maine, usa. c. borealis prey include the predatory gastropod nucella lapillus l., the herbiv ...200717536702
mechanism of a plastic phenotypic response: predator-induced shell thickening in the intertidal gastropod littorina obtusata.phenotypic plasticity has been the object of considerable interest over the past several decades, but in few cases are mechanisms underlying plastic responses well understood. for example, it is unclear whether predator-induced changes in gastropod shell morphology represent an active physiological response or a by-product of reduced feeding. we address this question by manipulating feeding and growth of intertidal snails, littorina obtusata, using two approaches: (i) exposure to predation cues ...200717465912
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 analytic ...200717275825
the use of a metallothionein-like-proteins (mtlp) kinetic approach for metal bioavailability monitoring in dredged material.ecotoxicological effects associated with contaminants present in dredged material from spain were determined using a marine biotest based on the determination of metallothionein-like-protein concentrations (mtlps) in the shore crab carcinus maenas. intermoult female c. maenas were exposed in the laboratory to sediments from three spanish ports, ría de huelva (sw, spain), la coruña (nw, spain) and bahía de cádiz (sw, spain) per replicate during 21 days. hepatopancreas samples from crabs were take ...200717218013
effect of temperature on the spoilage rate of whole, unprocessed crabs: carcinus maenas, necora puber and cancer pagurus.shelf life of whole, initially live, crabs depended primarily on the storage conditions and the time at which death occurred. large differences in the time that individual crab species survived particular storage conditions resulted in wide variations in shelf-life. bacterial spoilage of carcinus maenas, necora puber and cancer pagurus, measured using aerobic plate counts, indicated that on ice at 4 degrees c whole unprocessed crabs had a shelf life approximately 9-11 days, at 4 degrees c approx ...200717189768
herbivore-induced infochemicals influence foraging behaviour in two intertidal predators.herbivore-induced defences appear ubiquitous across most biomes and habitats. yet the direct correlation between induced changes in host plant chemistry and the population dynamics of the herbivore remain untested in many systems. in plant-herbivore interactions in the terrestrial environment, indirect or tritrophic interactions appear a successful way in which changes in the host plant chemistry induced by prior herbivory can impact on herbivore populations via increased success of natural enem ...200717106720
positive darwinian selection on crustacean hyperglycemic hormone (chh) of the green shore crab, carcinus maenas.the tissue-specific expression and differential function of the crustacean hyperglycemic hormone (chh) in carcinus maenas indicate an interesting evolutionary history. previous studies have shown that chh from the sinus gland x-organ (xo-type) has hyperglycemic activity, whereas the chh from the pericardial organ (po-type) neither shows hyperglycemic activity nor it inhibits y-organ ecdysteroid synthesis. here we examined the types of selective pressures operating on the variants of chh in carci ...200718391229
species invasion shifts the importance of predator dependence.the strength of interference between foraging individuals can influence per capita consumption rates, with important consequences for predator and prey populations and system stability. here we demonstrate how the replacement of a previously established invader, the predatory crab carcinus maenas, by the recently invading predatory crab hemigrapsus sanguineus shifts predation from a species that experiences strong predator interference (strong predator dependence) to one that experiences weak pr ...200718229836
molt-inhibiting hormone from the tropical land crab, gecarcinus lateralis: cloning, tissue expression, and expression of biologically active recombinant peptide in yeast.molt-inhibiting hormone (mih), a member of the crustacean hyperglycemic neuropeptide hormone family, inhibits ecdysteroidogenesis in the molting gland or y-organ (yo). a cdna encoding mih of the land crab (gel-mih) was cloned from eyestalk ganglia (eg) by a combination of reverse transcriptase polymerase chain reaction (rt-pcr) and 3'- and 5'-rapid amplification of cdna ends (race). the cdna (1.4 kb) encoded mih prohormone containing a 35 amino acid signal peptide and a 78 amino acid mature pept ...200717094991
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 glyc ...200717289410
antilipopolysaccharide factor (alf) of mud crab scylla paramamosain: molecular cloning, genomic organization and the antimicrobial activity of its synthetic lps binding domain.antilipopolysaccharide factors (alfs) are small basic proteins that can bind and neutralize lipopolysaccharide (lps) and have broad spectrum antimicrobial activities. in this study, we describe the isolation of the full-length cdna encoding for alf peptide (alfsp) of mud crab, scylla paramamosain by sequencing a hemocyte cdna library and using the rapid amplification cdna end (race) method. a full-length alfsp cdna of 614 bp contains an open reading frame (orf) of 372 bp, encoding 123 amino acid ...200717368541
molecular characterization and cell-specific expression of an ion transport peptide in the tobacco hornworm, manduca sexta.the crustacean hyperglycemic hormone (chh) peptides regulate diverse physiological processes from reproduction to metabolism and molting in arthropods. in insects, the ion transport peptides (itp), also members of the chh family, have only been implicated in ion transport. in this study, we sequenced a nucleotide fragment spanning the conserved a1/a2 region of the putative chh/itp gene. this fragment was amplified from larval cdna of the tobacco hornworm, manduca sexta and showed a high degree o ...200717450383
biomarkers study for sediment quality assessment in spanish ports using the crab carcinus maenas and the clam ruditapes philippinarum.intermolt crab carcinus maenas and clam ruditapes philippinarum were used to determine the toxicity of sediments collected in four spanish ports (cádiz, huelva, pasajes, and bilbao) under laboratory conditions during 28 days. sediment samples were analyzed to determine chemical concentration of metals (as, cd, cr, cu, fe, hg, mn, ni, pb, and zn), polyaromatic hydrocarbons, polychlorinated biphenyls, grain size distribution, and organic matter content. different biomarkers of exposure of early bi ...200717502980
amino acid sequence of crustacean hyperglycemic hormone (chh) from the crayfish, orconectes limosus: emergence of a novel neuropeptide family.the primary structure of the major form of chh from sinus glands of the crayfish, orconectes limosus, was determined by manual edman microsequencing. it is a 72-residue peptide with a calculated mr of 8400 da. in the number of residues, it is identical to the chh of carcinus maenas and very similar to mih (moult inhibiting hormone) of homarus americanus. all three peptides have pglu as n-terminus in common, and val-nh2 is the c-terminal residue in orconectes and carcinus chh. six cys residues oc ...20071800954
enzymatic biomarkers in the crab carcinus maenas from the minho river estuary (nw portugal) exposed to zinc and mercury.zinc and mercury are common contaminants of estuaries and coastal ecosystems where they may induce adverse effects on the biota. carcinus maenas is a key-species in several european estuaries, living in close association with the sediment where a considerable number of environmental contaminants, including zinc and mercury may accumulate. in the present study, the acute effects of zinc and mercury on c. maenas from the minho river estuary (nw portugal) were investigated by using the activity of ...200716949639
clarification of cercaria sevillana (digenea: microphallidae) life cycle using morphological and molecular data.cercaria sevillana is the cercaria larval stage that infects the gonads and the digestive gland of its first intermediate host, nassarius reticulatus. in this study the decapodous crustacean carcinus maenas was used to determine if it would serve as second intermediate host in the parasite's life cycle. the latter hypothesis was based on the knowledge that c. maenas is the second intermediate host of several other digenean species. after dissection, it was possible to observe encysted metacercar ...200717539414
a novel bacterial disease of the european shore crab, carcinus maenas molecular pathology and epidemiology.several rickettsia-like diseases have been reported in arthropods (insects and crustaceans), some of which result in significant losses of economically important species such as shrimp and crabs. this study reports on the molecular pathology of a recently emerged disease of the european shore crab, carcinus maenas, termed milky disease - named as a result of the unusual milky appearance of the haemolymph (blood). this disease was more prevalent (>26 %) during summer months when the water tempera ...200717768229
structural confirmation and occurrence of azaspiracids in scandinavian brown crabs (cancer pagurus).in 2005 and 2006, azaspiracids were for the first time detected in brown crabs (cancer pagurus) from the west coast of sweden and the north and north-west coast of norway. azaspiracids are marine toxins that have been detected in blue mussels in europe in recent years. on some occasions, they have been responsible for human intoxications with symptoms similar to those occurring by consumption of shellfish contaminated with okadaic acid group toxins. while the latter toxin group has been reported ...200817936866
the role of biomarkers to assess oil-contaminated sediment quality using toxicity tests with clams and crabs.a 28-d bioassay was conducted with two invertebrate species with different feeding habits, the clam ruditapes philippinarum and the shore crab carcinus maenas. the purpose of the present study was to assess the quality of sediments affected by oil spills in different areas of the spanish coast. the organisms were exposed to environmental samples of oil-contaminated sediments during four weeks and, after the experiment, a suite of biomarkers of exposure was measured: the phase one detoxification ...200818198936
field validation of a battery of biomarkers to assess sediment quality in spanish ports.two marine invertebrates, the crab carcinus maenas and the clam ruditapes philippinarum, were used as bioindicator species to assess contamination when exposed in situ to sediment from different sites from four spanish ports cadiz (sw spain), huelva (sw spain), bilbao (ne spain) and pasajes (ne spain). in an attempt to determine sediments toxicity, a combination of exposure biomarkers was analyzed in both species: metallothionein-like-proteins (mtlps), ethoxyresorufin o-deethylase (erod), glutat ...200817582664
accumulation and histopathological damage in the clam ruditapes philippinarum and the crab carcinus maenas to assess sediment toxicity in spanish ports.the degree of contamination and toxicity in sediment from four spanish ports (cádiz, huelva, pasajes and bilbao) was assessed in the present study. two marine invertebrate species, the shore crab carcinus maenas and the clam ruditapes philippinarum, were exposed to the different sediments under laboratory conditions for 28 days. relationships were developed among metal contamination (as, cd, cr, cu, fe, hg, mn, ni, pb and zn) in sediments, metal accumulation in gill tissues and histopathological ...200818313100
identification of putative crustacean neuropeptides using in silico analyses of publicly accessible expressed sequence tags.the development of expressed sequence tags (ests) for crustacean cdna libraries and their deposition in publicly accessible databases has generated a rich resource for peptide discovery in this commercially and ecologically important arthropod subphylum. here, we have conducted in silico searches of these databases for unannotated ests encoding putative neuropeptide precursors using the blast program tblastn, and have predicted the mature forms of the peptides encoded by them. the primary strate ...200818321503
crustin expression following bacterial injection and temperature change in the shore crab, carcinus maenas.the expression of carcinin, a crustin-type antimicrobial protein, in the crab, carcinus maenas, was studied following in vivo challenge with planococcus citreus, a gram-positive bacterium known to be killed by the encoded protein. real-time pcr analyses reveal that injection of p. citreus failed to elicit any significant changes in expression at 0-24h post-injection although there was a small, but significant, down-regulation at 84h in crabs held at 15 degrees c but not those at 5 or 20 degrees ...200818343497
toxicokinetic approach for the assessment of endocrine disruption effects of contaminated dredged material using female carcinus maenas.ecotoxicological effects associated with contaminants present in dredged material from three spanish ports, ría of huelva (sw, spain), la coruña (nw, spain) and bay of cádiz (sw, spain) were determined using a marine biotest based on endocrine disruption effects (vitellogenesis process alteration). intermoult female carcinus maenas were exposed in the laboratory to sediments from the spanish ports per replicate during 21 days. crab haemolymph samples were taken for vitellogenin/vitellin (vtg) an ...200818398677
isolation and characterisation of two antimicrobial peptides from haemocytes of the american lobster homarus americanus.two antimicrobial peptides from haemocytes of the american lobster, homarus americanus h. milne edwards 1837, were isolated and partially characterised - the first such description for this species. cap-2, an approximately 12kda peptide, contained amino acid sequences corresponding to the predicted sequence for hoa-crustin. crustins are whey acidic protein (wap) domain - containing peptides isolated from crustacean haemocytes. cap-2 did not have any activity towards the gram positive coccus aero ...200818499476
experimental evidence for latent developmental plasticity: intertidal whelks respond to a native but not an introduced predator.animals with highly inducible traits may show no inducible response when exposed to a related but wholly novel cue. this appears to be true for the intertidal whelk nucella lamellosa faced with a voracious introduced predator. in the laboratory, we exposed whelks to effluent from two species of predatory crab, the native red rock crab cancer productus and the invasive european green crab carcinus maenas. nucella and cancer have a long shared history in the northeast pacific, whereas potential in ...200818522925
[biosynthesis and metabolism of histamine in the central nervous system of carcinus maenas].the central nervous system of carcinus maenas synthesizes radioactive histamine when incubated in the presence of [14c] histidine and pyridoxal-5' phosphate. this biosynthesis increases linearly as a function of the amount of enzyme and the incubation time. it is not effected by heart, muscle or hepatopancreas extracts nor by haemolymph. thus histamine appears to be synthesized mainly in the nervous system. the latter is also the seat of carcinine (beta-alanylhistamine) biosynthesis. since carci ...20082441674
pattern and pathways for mercury lifespan bioaccumulation in carcinus maenas.carcinus maenas is an important and exploited natural resource in temperate estuaries, being consumed by local populations and also used as fish bait. through bioaccumulation and bioamplification processes occurring in polluted estuarine ecosystems, this species may directly or indirectly represent a major pathway for human contamination. no information is available on the overall lifespan bioaccumulation pattern of mercury for this species, which would be an invaluable tool for assessing the ri ...200818455199
identification and developmental expression of mrnas encoding putative insect cuticle hardening hormone, bursicon in the green shore crab carcinus maenas.bursicon is the ultimate hormone in insect ecdysis, which is involved in cuticle hardening. here we show that mrnas encoding the heterodimeric cystine knot protein bursicon (burs alpha, beta), are present in crustaceans, suggesting ubiquity of this hormone in arthropods. we firstly report the cloning, sequencing of mrnas encoding subunits from the water flea, daphnia arenata and the cns of the crab, carcinus maenas, in comparison with insect bursicon subunits. expression patterns of alpha and be ...200818221939
effects of 4-nonylphenol on the endocrine system of the shore crab, carcinus maenas.there is a considerable body of evidence to suggest that many anthropogenic chemicals, most notably xeno-estrogens, are able to disrupt the endocrine system of vertebrates. there have been few comparable studies on the effects of exposure to these chemicals that may serve as biomarkers of endocrine disruption in aquatic invertebrate species. in addition, the evidence available is complex, conflicting, and far from conclusive. the present study aimed to investigate the impact of the xeno-estrogen ...200818214899
inhibition between invasives: a newly introduced predator moderates the impacts of a previously established invasive predator.1. with continued globalization, species are being transported and introduced into novel habitats at an accelerating rate. interactions between invasive species may provide important mechanisms that moderate their impacts on native species. 2. the european green crab carcinus maenas is an aggressive predator that was introduced to the east coast of north america in the mid-1800 s and is capable of rapid consumption of bivalve prey. a newer invasive predator, the asian shore crab hemigrapsus sang ...200818177327
skeletal adaptations for forwards and sideways walking in three species of decapod crustaceans.crustaceans have been successfully employed to study legged locomotion for decades. most studies have focused on either forwards-walking macrurans, or sideways-walking brachyurans. libinia emarginata is a majoid crab (brachyura) and as such belongs to the earliest group to have evolved the crab form from homoloid ancestors. unlike most brachyurans, libinia walks forwards 80% of the time. we employed standard anatomical techniques and motion analysis to compare the skeleton, stance, and the range ...200818089130
accumulation and depuration of okadaic acid esters in the european green crab (carcinus maenas) during a feeding study.soft shell crab is a seafood delicacy in many parts of the world. in denmark, it has been investigated whether a commercial production of soft shell european green crabs (carcinus maenas) would be feasible. in relation to this, a feeding study was performed to examine if occurrence of dsp toxins in the product could be a food safety problem. the crabs were fed with mussels containing dsp toxins (2500 microg total okadaic acid equivalents/kg) for 17 days and then fasted for 19 days. the content o ...200817983637
influence of predator density on nonindependent effects of multiple predator species.interactions between multiple predator species are frequent in natural communities and can have important implications for shared prey survival. predator density may be an important component of these interactions between predator species, as the frequency of interactions between species is largely determined by species density. here we experimentally examine the importance of predator density for interactions between predator species and subsequent impacts on prey. we show that aggressive inter ...200817968591
thermodynamics of oxygenation-linked proton and lactate binding govern the temperature sensitivity of o2 binding in crustacean (carcinus maenas) hemocyanin.with the aim of understanding the molecular underpinnings of the enormous variation in the temperature sensitivity of hemocyanin-o2 affinity encountered in crustaceans, we measured o2 binding to carcinus maenas hemocyanin at two temperatures, varying ph values and in the absence and presence of lactate ions in order to assess the contributions of oxygenation-linked binding of protons (the bohr effect) and lactate ions to the overall enthalpies of oxygenation (deltah'). the hemocyanin binds maxim ...200818344479
permanent genetic resources: isolation and characterization of microsatellite loci in the mediterranean shore crab carcinus aestuarii (decapoda, portunidae).we characterized nine polymorphic microsatellites in the mediterranean shore crab carcinus aestuarii (decapoda: portunidae). microsatellites were isolated from a partial genomic library enriched for multiple motifs. all loci were polymorphic, with number of alleles ranging from two to 16 and a mean observed heterozygosity of 0.75. seven loci were in hardy-weinberg equilibrium, and two showed weak heterozygote deficiency. no linkage disequilibrium was found between loci. in addition, we tested ha ...200821585795
differential snail predation by an exotic crab and the geography of shell-claw covariance in the northwest atlantic.here we investigate if predation by the european green crab (carcinus maenas) differs between two congeneric snails in the northwest atlantic (littorina littorea and l. obtusata), and ask if differential predation can help explain the geography of claw and shell forms among geographically separated populations. first, correlations between crusher-claw size and shell mass -- tested across a wide size range of animals -- were highly significant among populations of c. maenas and l. obtusata, where ...200818298647
nutritional status of carcinus maenas (crustacea: decapoda) influences susceptibility to contaminant exposure.using the shore crab carcinus maenas as a model, this study tested the hypothesis that nutritional status influences susceptibility of adult crabs (>60mm carapace width (cw)) to environmental contamination. in the laboratory, crabs were either starved, given a restricted diet (fed on alternate days) or fully fed (fed each day). in addition, crabs under each feeding regime were exposed to a sublethal concentration (200microgl(-1)) of pyrene (pyr) as a model organic (pah (polyaromatic hydrocarbon) ...200818606465
ambient fauna impairs parasite transmission in a marine parasite-host system.to understand possible factors controlling transmission of trematode larvae between first and second intermediate hosts we examined the impact of ambient fauna on parasite transmission in a marine intertidal parasite-host association. cockle hosts (cerastoderma edule) kept together with selected co-occurring macrozoobenthic species in mesocosms acquired a lower parasite load compared to cockles kept alone, when targeted by cercariae of the trematode himasthla elongata. the reduction of parasite ...200818561867
differential expression and induction of two carbonic anhydrase isoforms in the gills of the euryhaline green crab, carcinus maenas, in response to low salinity.two isoforms of the enzyme carbonic anhydrase (ca) from the gills of the euryhaline green crab were sequenced and identified; these were found to match the cytoplasmic (cac) and membrane-associated (cag) isoforms known from other species. the mrna of the membrane-associated isoform is present in significantly higher levels of abundance in gills of crabs acclimated to 32 ppt, at which the crab is an osmotic and ionic conformer. upon transfer to low salinity (15 ppt), in which the crab is an osmor ...200820483218
angiotensin-converting enzyme-like activity in crab gills and its putative role in degradation of crustacean hyperglycemic hormone.angiotensin-converting enzyme-like enzyme activity (acela) was found in carcinus maenas using reverse phase high performance liquid chromatography (rp-hplc) analysis of degradation kinetics of a synthetic substrate (hippuryl-histidyl-leucine) and a specific inhibitor (captopril). gills contained the highest acela, then brain, muscle, and testis, respectively, while no activity was detected in the following tissues: hepatopancreas, hindgut, hypodermis, heart, and hemolymph. acela present in gill ...200818481304
a robotic molecular method for in situ detection of marine invertebrate larvae.knowledge of the temporal and spatial abundance of invertebrate larvae is critical to understanding the dispersal capabilities and recruitment potential of marine and aquatic organisms. traditional microscopic analyses are time-consuming and difficult given the diversity of larval species and a frequent lack of discriminating morphological characteristics. here, we describe a sensitive rrna targeted sandwich hybridization assay (sha) that uses oligonucleotide probes to detect and enumerate the l ...200821585831
sublethal responses in caged organisms exposed to sediments affected by oil spills.this study was performed to determine sublethal responses of two invertebrate species by using field deployments in areas affected by oil spills, which are acute in the galician coast (nnw, spain) and chronic in the bay of algeciras (ssw, spain). the organisms employed were the crab carcinus maenas and the clam ruditapes philippinarum, and during 28 days the animals were exposed to contaminated sediments in cages under field conditions. different biomarkers of exposure were determined after a 28 ...200818420252
genetic patterns across multiple introductions of the globally invasive crab genus carcinus.the european green crab carcinus maenas is one of the world's most successful aquatic invaders, having established populations on every continent with temperate shores. here we describe patterns of genetic diversity across both the native and introduced ranges of c. maenas and its sister species, c. aestuarii, including all known non-native populations. the global data set includes sequences from the mitochondrial cytochrome c oxidase subunit i gene, as well as multilocus genotype data from nine ...200819120987
quantification of metallothionein by differential pulse polarography overestimates concentrations in crustaceans.if metallothionein concentrations in invertebrates are to be used as biomarkers for metal contamination in the aquatic environment, it is imperative thatthe methods used for quantitative analysis are reliable. a review of the literature concerned with quantification of crustacean metallothionein shows that utilization of differential pulse polarography generally results in higher concentrations than any other method. the obvious discrepancies were investigated by experimental comparison of three ...200819068828
cytochrome p450 expression is moult stage specific and regulated by ecdysteroids and xenobiotics in the crab carcinus maenas.the shore crab carcinus maenas has a high capacity for metabolizing polycyclic aromatic hydrocarbons (pahs). cytochrome p450 (cyp) enzymes are involved in this metabolism and also have a role in development and reproduction. this investigation is a systematic gene expression analysis of six cyps in c. maenas. expression of cyp2 and cyp3-like genes was predominant in hepatopancreas, while expression of cyp4-like genes was predominant in gills and epidermis. expression of all six cyp genes fluctua ...200818983823
resource identity modifies the influence of predation risk on ecosystem function.it is well established that predators can scare as well as consume their prey. in many systems, the fear of being eaten causes trait-mediated cascades whose strength can rival or exceed that of more widely recognized density-mediated cascades transmitted by predators that consume their prey. despite this progress it is only beginning to be understood how the influence of predation risk is shaped by environmental context and whether it can exert an important influence on ecosystem-level processes ...200818959317
non-destructive assessment of polycyclic aromatic hydrocarbon (pah) exposure by fluorimetric analysis of crab urine.the detection of urinary polycyclic aromatic hydrocarbon (pah) metabolites by fluorescence spectrophotometry is particularly effective as a practical means to assess pah exposure in decapod crabs. however, the practical application of this technique has thus far only been tested for the european shore crab (carcinus maenas) and only a few field studies have been conducted in heavily polluted areas. the present study evaluated the adaptability of this method as a rapid, cost-effective and non-des ...200818845307
degree of neuromuscular facilitation is correlated with contribution to walking in leg muscles of two species of crab.despite decades of work on the neuromuscular physiology of crustacean leg muscles, little is known about how physiological differences between these muscles relate to their behavioral usage. we studied a sideways walking shore crab, carcinus maenas, and a forward walking spider crab, libinia emarginata, as part of our work to understand the neural control of locomotion. the two species differed significantly in facilitation at neuromuscular junctions for every muscle studied. further, these diff ...200818830606
physiological responses of juvenile and adult shore crabs carcinus maenas (crustacea: decapoda) to pyrene exposure.this study measured aspects of the physiology of juvenile [(<35mm carapace width (cw)] and adult (>60mm cw) carcinus maenas to test the hypothesis that these different life-history stages exhibit different sensitivities to environmental contamination. newly-collected juveniles had significantly (p<0.05) lower immunocompetence (phagocytosis and cellular integrity), lower metabolic energy (haemolymph glucose) and increased scope for growth compared with adults. seven day exposure to a sub-lethal c ...200818774603
methyl farnesoate couples environmental changes to testicular development in the crab carcinus maenas.carcinus maenas males have two major color phases. green-phase males molt frequently and tend to live in brackish estuaries during the summer. after becoming red-phase males, they molt infrequently, have higher mating success, and live in cooler, deeper water. we found profound differences between these two phases in the way salinity and temperature affect hemolymph levels of methyl farnesoate (mf), a hormone that affects crustacean reproduction. few green-phase males (<10%) had detectable mf in ...200818723534
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. histolog ...200818714687
[conditioned food reflexes from the visual, tactile and static receptors of the black sea crab carcinus maenas]. 200813962447
acute effects of chlorpyryphos-ethyl and secondary treated effluents on acetylcholinesterase and butyrylcholinesterase activities in carcinus maenas.the acute effects of commercial formulation of chlorpyrifos-ethyl (dursban) and the secondary treated industrial/urban effluent (stiue) exposure on acetylcholinesterase (ache) and butyrylcholinesterase (buche) activities in hepatopancreas and gills of mediterranean crab carcinus maenas were investigated. after 2 d of exposure to chlorpyriphos-ethyl, the ache activity was inhibited in both organs at concentrations of 3.12 and 7.82 microg/l, whereas the buche was inhibited only at higher concentra ...200920000004
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