Seasonal variations in the biochemical composition of the crayfish Parastacus defossus (Crustacea, Decapoda) in its natural environment.

The crayfish Parastacus defossus occurs in Uruguay and the state of Rio Grande do Sul in Brazil. It lives in swamps and lakes, where it digs sloped subterranean tunnels that are used as burrows. Because there is little information about the biology, physiology and ecology of this species, the aim of this study was to identify the seasonal variations of its carbohydrate and lipid metabolism. Crayfish were collected monthly (from November 2002 to February 2004) in the Lami region, Porto Alegre municipality (30 degrees 11'41'S-51 degrees 06'00'W). Haemolymph samples, used for determination of glucose, total proteins, triglycerides, total cholesterol and total lipids, were collected in the field using potassium oxalate as an anti-clotting agent. The animals and haemolymph samples were immediately frozen in the field. In the laboratory, the hepatopancreas, gills and abdominal muscles were removed for determination of glycogen, triglycerides, total cholesterol and total lipids. The findings suggest that in P. defossus, lipids are an important reserve of energy used during reproduction in both males and females; whereas glycogen may be used during periods of intense activity or environmental stress.     

Seasonal variations in the intermediate metabolism of Parastacus varicosus (Crustacea, Decapoda, Parastacidae)

The aim of this study was to analyze the seasonal variations in the metabolism of Parastacus varicosus and examine the possible relationships to its reproduction. Animals were sampled (9 h to 10 h) in each month in the Gravataí River, RS, Brazil. Haemolymph samples were collected from each crayfish in the field for determination of glucose, total proteins, total lipids, and total cholesterol. Hepatopancreas, abdominal muscle, and gonads were removed for determination of glycogen, total proteins, total lipids, and total cholesterol. ANOVA revealed significant seasonal differences in the biochemical composition of all tissues studied; when the sexes were compared these parameters did not show any significant difference in the hepatopancreas and muscle. However, in haemolymph we observed significant variation only in cholesterol and lipid levels. The results suggest that the metabolic variability is related to the stage of maturation of the gonads, in females, where the hepatopancreas and other tissue studies can store and transfer reserves to support maturation to complement the food intake. Variations in the gonadosomatic and hepatosomatic indices suggest that reproduction occurs principally in summer. As in other decapods, abiotic factors such as water temperature, oxygen content, etc. influence the intermediate metabolism.     

Metabolic responses in two species of crayfish (Parastacus defossus and Parastacus brasiliensis) to post-hypoxia recovery

The period of post-hypoxia recovery is essential for the rapid replenishment of energy reserves and for the removal of metabolic end products formed during hypoxia. Periods of post-hypoxia recovery were analyzed in two crayfish species, where Parastacus defossus is a fossorial species, and Parastacus brasiliensis lives in lotic environments with higher oxygen levels. After 4 h of hypoxia (2 mg O(2)/L), groups of animals were placed in tanks with oxygenated water and were then removed at intervals of 1, 3, 6, and 9 h. Hemolymph and tissues (hepatopancreas, muscle, and anterior and posterior gills) were extracted for the determination of glucose, lactate, free glucose, glycogen, total proteins, total lipids, arginine phosphate, and arginine. As expected, lactate levels were restored more rapidly in P. defossus than in P. brasiliensis. P. defossus restored its glycogen reserves of the hepatopancreas and muscle tissue. Free glucose was quickly restored in all tissues of both species. In relation to arginine phosphate reserves, P. defossus showed a greater ability to restore this metabolite in the hepatopancreas. Both species recovered their arginine phosphate reserves, but they also used this metabolite in longer periods of recovery. Mainly in P. brasiliensis the reserves of total lipids seem to be an important source of energy during the recovery period. The animals developed various metabolic strategies to post-hypoxia recovery, mainly P. defossus which restored its reserves more completely and more rapidly than did P. brasiliensis.     

Seasonal variations in the biochemical composition and lipoperoxidation of Hyalella curvispina (Crustacea, Amphipoda)

We assessed the effect of seasonal variations on the biochemical composition and lipoperoxidation of an amphipod crustacean, Hyalella curvispina, which inhabits the coastal plain of the state of Rio Grande do Sul, and correlated this with information on the biology of the species. The animals were collected monthly from September 2003 to August 2005, in Gentil Lagoon, Tramandaí, Rio Grande do Sul, Brazil. Glycogen levels, total proteins, total lipids, triglycerides, and lipoperoxidation were measured for each sex separately. The results suggest that these animals have seasonal storage and degradation of the energy substrates investigated. During summer, glycogen, total lipids, and triglycerides were stored, and were utilized especially during autumn and winter. Proteins were utilized differently in males and females. Females stored proteins especially in spring and used them in summer; whereas males stored proteins especially in spring and used them in winter. The levels of lipoperoxidation during the year differed between females and males. These variations may be related to biotic factors (e.g., reproduction period and others) and to abiotic factors (e.g., water temperature, salinity and others).     

Characterization of the sexual pattern of Parastacus varicosus (Crustacea: Decapoda: Parastacidae)

Abstract. Freshwater crayfishes of the genus Parastacus are intersex, i.e., show characteristics of both sexes in the same individual; also, intersexuality has been documented in hermaphroditic species. The aim of this study was to analyze the gonads of Parastacus varicosus , characterizing its sexual pattern. The animals were collected at Cova do Touro, Gravataí, Rio Grande do Sul, Brazil. Three sexual forms were identified: intersex females, intersex males, and transitional specimens showing an ovotestes gonad that could only be identified by means of histological analysis. All specimens had two pairs of gonopores and gonoducts. The oviduct had a wider diameter in females, whereas in males the vasa deferentia were more developed. Gonads were composed of two parallel structures located in the cephalothorax. Female gonads were classified into three stages. The presence of transitional specimens may indicate that sex change occurs, with protandric hermaphroditism, as observed in other species of Parastacus . Because previous studies had demonstrated that P. varicosus is a gonochoristic species in a population from Uruguay, the present study addressed the possibility that the sexual pattern depends on differences in environmental factors among populations. To improve understanding of evolution of hermaphroditism in these crustaceans, their reproductive dynamics should be studied, including identification of the factors stimulating male and female functions.     

Constant morphological patterns in the hemolymph vascular system of crayfish (Crustacea,Decapoda)

We present a study of the hemolymph vascular system of the marbled crayfish, Procambarus fallax f. virginalis, the only crayfish species known to be parthenogenetic. To identify potential evolutionary patterns, we compared data from a total of 48 specimens of P. fallax with 22 specimens of Orconectes limosus. Visualizations (2D and 3D) were carried out using a combination of classical and modern morphological techniques. Our data were compared to the existing literature.Like all Decapoda, both P. fallax and O. limosus have a hemolymph vascular system, consisting of a globular heart with seven off-branching arteries. We were able to visualize in detail the heart of crayfish for the first time, i.e., the myocard with its clusters of muscles running through the lumen of the heart, the valves and flaps of ostia and arteries. Furthermore, the branching patterns of the seven artery systems were analyzed. Anatomical structures identified to be consistent in all specimens of both species were combined as ground pattern of hemolymph vascular system features for Astacida.     

Seasonal variations in larval biomass and biochemical composition of brown shrimp, Crangon crangon (Decapoda, Caridea), at hatching

The “brown shrimp”, Crangon crangon (Linnaeus 1758), is a benthic key species in the North Sea ecosystem, supporting an intense commercial fishery. Its reproductive pattern is characterized by a continuous spawning season from mid-winter to early autumn. During this extended period, C. crangon shows significant seasonal variations in egg size and embryonic biomass, which may influence larval quality at hatching. In the present study, we quantified seasonal changes in dry weight (W) and chemical composition (CHN, protein and lipid) of newly hatched larvae of C. crangon. Our data revealed significant variations, with maximum biomass values at the beginning of the hatching season (February–March), a decrease throughout spring (April–May) and a minimum in summer (June–September). While all absolute values of biomass and biochemical constituents per larva showed highly significant differences between months (P < 0.001), CHN, protein and lipid concentrations (expressed as percentage values of dry weight) showed only marginally significant differences (P < 0.05). According to generalized additive models (GAM), key variables of embryonic development exerted significant effects on larval condition at hatching: The larval carbon content (C) was positively correlated with embryonic carbon content shortly after egg-laying (r ² = 0.60; P < 0.001) and negatively with the average incubation temperature during the period of embryonic development (r ² = 0.35; P < 0.001). Additionally, water temperature (r ² = 0.57; P < 0.001) and food availability (phytoplankton C; r ² = 0.39; P < 0.001) at the time of hatching were negatively correlated with larval C content at hatching. In conclusion, “winter larvae” hatching from larger “winter eggs” showed higher initial values of biomass compared to “summer larvae” originating from smaller “summer eggs”. This indicates carry-over effects persisting from the embryonic to the larval phase. Since “winter larvae” are more likely exposed to poor nutritional conditions, intraspecific variability in larval biomass at hatching is interpreted as part of an adaptive reproductive strategy compensating for strong seasonality in plankton production and transitory periods of larval food limitation.     

Tyrosine hydroxylase activity in the central nervous system of the crayfish,Pacifastacus leniusculus (Crustacea,Decapoda)

Summary Tyrosine hydroxylase, responsible for the formation ofl-dopa froml-tyrosine, has been identified in the central nervous system of the crayfish,Pacifastacus leniusculus (Crustacea, Decapoda). It requires pterine as cofactor and is inhibited by a number of known tyrosine hydroxylase inhibitors; iron-chelators, tyrosine analogues and also by the catecholamines, dopamine and noradrenaline. Iron enhances the activity of the enzyme. It differs from the vertebrate tyrosine hydroxylase in having a more alkaline pH optimum and a higher affinity for the pterine cofactor. Kinetic studies were performed andK _m andV _max values are presented. Dopa formed was identified and quantitatively measured by high pressure liquid chromatography (HPLC) and electrochemical detection.     

Embryonic expression patterns of the Hox genes of the crayfish Procambarus clarkii (Crustacea, Decapoda)

SUMMARY Higher crustaceans (class Malacostraca) represent the most species-rich and morphologically diverse group of non-insect arthropods. The superorders Eucarida and Peracarida, two large groups that separated over 350 million years ago, encompass most malacostracan diversity. Recently, the Hox genes of the peracarid woodlouse Porcellio scaber (Isopoda) were shown to be expressed in domains that coincide with morphological boundaries of body tagmata, which differ from those in insects ( Abzhanov and Kaufman 1999a,b ). Moreover, observed changes in Hox expression domains during ontogeny correlate with morphological remodeling, such as a transformation of the first thoracic leg into mouthpart maxillipeds, which occurs in the trunk of the embryo. Decapods have a different modification of the malacostracan bodyplan, with up to three pairs of maxillipeds and extensive fusion and cephalization of the thorax. Here we describe expression patterns of the trunk Hox genes Scr, Antp, Ubx, abd-A and cad in the eucarid crayfish Procambarus clarkii (Decapoda). We find that the crayfish expression patterns, for the most part, resemble those of the woodlouse Porcellio scaber (Isopoda), but are more modulated and complex . Nevertheless, as in Porcellio the boundaries of the Hox expression domains do correlate with morphological features and their modulations to transformations in the embryo. Thus we propose that the trunk Hox genes were likely important in the evolution of and currently play an essential role in the development of the complex decapod bodyplan.     

Seasonal variations in the total lipid content of Chorismus antarcticus (Pfeffer) (Crustacea: Decapoda) at South Georgia

The Antarctic benthic prawn Chorismus antarcticus (Pfeffer) was sampled regularly for a year and its total lipid content measured. Immature and transitional prawns had very variable lipid contents but similar mean values (2.92 and 2.91 % fresh wt, respectively); the variation was, in part, due to fluctuations in lipid reserves associated with ecdysis. Maturation of the testes involved the synthesis of ≈ 1.8 mg lipid, and female prawns synthesized an average of 50 % of their own lipid content for the gonad. Fecundity did not appear to be limited by the demand for egg lipid and synchrony in the breeding cycle was probably related to the planktonic feeding of the newly-hatched larvae. Eggs showed an unusual increase in total lipid content during the early stages of embryonic development, followed by the more normal decrease; larvae probably hatch with some reserve lipid.Available data on lipid contents of Antarctic crustaceans are briefly reviewed. The high lipid contents observed in herbivorous Antarctic planktonic crustaceans are due to the patterns of feeding imposed by the strongly seasonal nature of primary production in Antarctic waters, rather than low temperature per se. There is no evidence from the benthos of any systematic elevation of lipid content in polar species, although there is an indication of an increased lipid content in the muscle of polar crustaceans.     

Seasonal variations in the intermediate metabolism of Aegla platensis (Crustacea, Aeglidae)

This study investigated the effects of circadian and seasonal fluctuations on the intermediate metabolism of the freshwater crustacean Aegla platensis. Individuals were collected each month, at 6:00 h, 12:00 h and 18:00 h, between August 1999 and June 2002 in the Municipality of Taquara, Rio Grande do Sul, Brazil (29 degrees 30'0.2'S and 50 degrees 46'50'W). Anomuran crabs were separated by sex, and samples of haemolymph, hepatopancreas, abdominal muscle, and gills were collected for quantification of glucose, proteins, lipids, triglycerides, and glycogen. No variations were observed in the analyzed metabolic parameters for the different tissues during the different times of day, either for males or females. However, the storage and use of the energetic substrates analyzed did fluctuate seasonally. These results suggest an increase in energy demand, possibly for the production of gametes during summer, incubation and egg laying during autumn and winter, and parental care during spring and summer, respectively. A similar pattern has been observed for other crustaceans.     

Directional selectivities of near-field filiform hair mechanoreceptors on the crayfish tailfan (Crustacea: Decapoda)

Directional selectivities of mechanoreceptors that innervate filiform hairs on the crayfish tailfan were investigated with unidirectional, sinusoidal, water-motion stimuli. These recordings provide the first representative sample from filiform hair sensilla on the entire tailfan. The filiform hair receptors exhibit unimodal directional selectivity patterns that were well fitted by a cardioid function with a half-width of 122°. The preferred directions correspond to the major axis of hair motion, and are perpendicular to the orientation of lateral branches on the main hair shaft. Pooled plots of preferred directions demonstrate quadrimodal patterns on the telson and endopods which are associated with hair location, and a bimodal pattern on the exopods. For each appendage, the combination of the overall pattern of preferred directions with “coarse coding” of direction by individual receptors provides sensitivity to a full 0–360° range of water motion and the potential to discriminate the direction of water motion throughout this range. The results demonstrate several similarities to the wind-sensitive cercal receptor system in orthopteroid insects, and suggest that crustacean filiform hair receptors provide a sufficient sensory basis for behavioral orientation to water currents and shorelines. Accepted: 5 January 1998     

Ontogeny of the antennal glands in the crayfish Astacus leptodactylus (Crustacea, Decapoda): anatomical and cell differentiation

The ontogeny of the antennal glands was studied during the embryonic and post-embryonic development of Astacus leptodactylus. The future glands arising from undifferentiated columnar cells were detectable at the metanauplius stage EI 150 m (EI: eye index; approximately 440 m at hatching). The tubule and labyrinth differentiated in embryos at EI 190 m, and the bladder and coelomosac at EI 250 m. At EI 350 m, the tubule lengthened and divided into proximal and distal sub-regions. In later stages, the gland retained the same morpho-anatomy but the differentiation and size of each part increased. The cells of the coelomosac displayed the cytological features of podocytes in late embryonic development at EI 440 m. Only small apical microvilli and a few mitochondria were observable in the labyrinth cells at EI 250 m; by EI 440 m, these cells presented well-shaped apical microvilli, formed bodies, basal infoldings and mitochondria. In the cells of the tubules and bladder, mitochondria and basal infoldings occurred at EI 440 m and EI 250 m, respectively. The differentiation of the tubules and bladder cells suggested that they were involved in active transport at EI 440 m. Following hatching, the differentiation of the cells and the size of the glands increased. The ontogeny of the antennal glands thus starts in early embryos, the specific cellular functional features being differentiated in the various parts of the glands by EI 440 m. The antennal glands are probably functional just before hatching, i.e., before the juveniles are confronted with the low osmolality of freshwater.Thanks are due to the University of Tarbiat Modarres and Ministry of Science, Research and Technology, Islamic Republic of Iran, for financial aid and support. Special thanks are also extended to the Société Française dExportation des Ressources Educatives (SFERE) for a scholarship to S.K.     

The effects of nitrite on chloride regulation in the crayfish Pacifastacus leniusculus Dana (Crustacea: Decapoda)

Summary Nitrite in the external freshwater medium was found to be toxic to Pacifastacus leniusculus Dana (48 h LC₅₀0.7 mM NO ₂ ^– ). It produced significant changes in haemolymph ionic concentration and acid-base status. Exposure to 1.0 mM NO ₂ ^– resulted in a rapid, active accumulation of nitrite in the haemolymph (to 25 mM NO ₂ ^– after 24 h) and caused the partial inhibition of Cl^– uptake. Some reduction in Cl^– efflux rate was seen. In 1.0 mM NO ₂ ^– a rapid depletion of haemolymph [Cl^–] was observed (50 mM decrease in 27 h). Nitrite competitively inhibited active Cl^– uptake (K_m increased from 0.42 to 1.22 mM; K_i=0.45 mM). To achieve Cl^– balance in this medium, depleted crayfish would require a two-fold increase in external [Cl^–]. A lesser decrease in haemolymph [Na⁺] was found while osmotic pressure was relatively unaffected. Haemolymph [HCO ₃ ^– ] showed a significant increase and was accompanied, unexpectedly, by an acidosis. Possible sources of the excess HCO ₃ ^– , perhaps by inhibition of normal Cl^–/HCO ₃ ^– branchial exchange or release from CaCO₃ stores, are discussed. Haemolymph clearance of NO ₂ ^– was slower than uptake as was the restoration of [Cl^–] on recovery in nitrite-free medium.Abbreviations AFWM artificial freshwater medium - BOD biochemical oxygen demand - J _out ^Cl chloride efflux - J _in ^Cl chloride influx - J _in ^Cl chloride influx - J _net ^base net base flux - J _net ^base net base flux - J _in(p) ^Cl passive chloride influx - J _out efflux - LC ₅₀ median lethal concentration - NEDE N-1-Naphthylethylenediamine - SEM standard error of mean - TEP transepithelial potential difference - V _in ^Cl active chloride uptake     

Phylogenetic systematics of the reptantian Decapoda (Crustacea, Malacostraca)

Although the biology of the reptantian Decapoda has been much studied, the last comprehensive review of reptantian systematics was published more than 80 years ago. We have used cladistic methods to reconstruct the phylogenetic system of the reptantian Decapoda. We can show that the Reptantia represent a monophyletic taxon. The classical groups, the 'Palinura', 'Astacura' and 'Anomura' are paraphyletic assemblages. The Polychelida is the sister-group of all other reptantians. The Astacida is not closely related to the Homarida, but is part of a large monophyletic taxon which also includes the Thalassinida, Anomala and Brachyura. The Anomala and Brachyura are sister-groups and the Thalassinida is the sister-group of both of them. Based on our reconstruction of the sister-group relationships within the Reptantia, we discuss alternative hypotheses of reptantian interrelationships, the systematic position of the Reptantia within the decapods, and draw some conclusions concerning the habits and appearance of the reptantian stem species.     

Evolution of the thaumastocheliform lobsters (Crustacea,Decapoda, Nephropidae)

Deep‐sea lobsters previously assigned to the family Thaumastochelidae Bate, 1888, the thaumastocheliforms, have very distinctive, greatly unequal first chelipeds, with the right side extremely elongate and pectinate, and in having short, quadrate pleonal pleura. Despite interesting morphology and a long taxonomic history, the phylogeny of the group has received little detailed analysis. Here, we conduct a species‐level phylogenetic analysis of the thaumastocheliforms based on morphological and molecular data (three mitochondrial genes: COI, 16S rDNA and 12S rDNA; two nuclear protein‐coding genes: H3 and NaK) to robustly reconstruct their evolutionary history and estimate divergence times. Separate and combined analyses of all data sources support thaumastocheliform monophyly, but as a clade deeply nested within the Nephropidae supporting recent synonymy of Thaumastochelidae with Nephropidae. Combined and molecular‐only analyses support generic monophyly of all three thaumastocheliform genera and Dinochelus as sister to Thaumastochelopsis, fully corroborating the current, morphology‐based taxonomy. In contrast, Thaumastocheles is recovered as paraphyletic in morphology‐only analyses owing to minimal character support. The Cretaceous–Paleogene Oncopareia was recovered as a stem‐lineage thaumastocheliform. The fossil record indicates that the thaumastocheliforms once lived in shallow, continental shelf depths, but moved into deeper water in the Cenozoic where they occur today. The thaumastocheliforms originated in northern Europe during the Mid‐Late Cretaceous and later dispersed westwards to the south‐eastern Pacific through the western Atlantic and eastwards to the western Pacific through the Indian Ocean. Thaumastochelopsis can be considered the most derived thaumastocheliform genus based on the degree of structural reduction relative to other thaumastocheliforms, its remote geographical occurrence (Australia) from the hypothesised place of origin (northern Europe) and its more recent estimated divergence than other genera (28 Mya for the MRCA of extant species of the genus).     

Cell lineage studies in the crayfish Cherax destructor (Crustacea,Decapoda) : germ band formation,segmentation, and early neurogenesis

Summary The cell division pattern of the germ band of Cherax destructor is described from gastrulation to segmentation, limb bud formation, and early neurogenesis. The naupliar segments are formed almost simultaneously from scattered ectoderm cells arranged in a V-shaped germ disc, anterior to the blastopore. No specific cell division pattern is recognisable. The post-naupliar segments are formed successively from front to rear. Most post-naupliar material is budded by a ring of about 39 to 46 ectoteloblasts, which are differentiated successively and in situ in front of the telson ectoderm. The ectoteloblasts give rise to 15 descendant cell rows by unequal divisions in an anterior direction, following a mediolateral mitotic wave. Scattered blastoderm cells of non-ectoteloblastic origin in front of the ectoteloblast descendants and behind the mandibular region are also arranged in rows. Despite their different origins, teloblastic and non-teloblastic rows cleave twice by mediolateral mitotic waves to form 4 regular descendant rows each. Thereafter, the resulting grid-like pattern is dissolved by stereotyped differential cleavages. Neuroblasts are formed during these differential cleavages and segmentation becomes visible. Each ectoderm row represents a parasegmental unit. Therefore, the segmental boundary lies within the area covered by the descendants of 1 row. Segmental structures (limbs, ganglia) are composed of derivatives of 2 ectoderm rows. The results are compared with the early development of other crustaceans and insects in relation to mechanisms of germ band formation, segmentation, neurogenesis, and evolution.