Diurnal variations in physiology and behaviour of the noble crayfish Astacus astacus and the signal crayfish Pacifastacus leniusculus

Heart rate, locomotor activity, and oxygen consumption were recorded simultaneously and continuously in seven individuals of the noble crayfish Astacus astacus (Linneus 1758) and seven individuals of the signal crayfish Pacifastacus leniusculus (Dana 1852). The recordings were made in the laboratory over 7 days at 15°C under a 12 : 12 h dark : light regime. Circadian rhythms in heart rate, locomotor activity and oxygen consumption were found both in A. astacus and P. leniusculus. Increased heart rate, locomotor activity, and oxygen consumption levels during night time in both A. astacus and P. leniusculus illustrated expression of nocturnal behaviour. No differences in oxygen consumption levels were observed between A. astacus and P. leniusculus. Also, no significant difference between heart rate levels or heart rate variances was found in A. astacus and P. leniusculus at night. During day, however, heart rate levels, heart rate variances and locomotor activity were higher in P. leniusculus than in A. astacus. The higher activity level in P. leniusculus than in A. astacus during daytime indicates that P. leniusculus is less strictly nocturnal than is A. astacus.     

Size scaling of whole‐body metabolic activity in the noble crayfish (Astacus astacus) estimated from measurements on a single leg

1. Use of electron transport system (ETS) activity in a single leg for estimating whole‐body ETS activity was explored in the noble crayfish Astacus astacus. Oxygen consumption and ETS activity of the whole body and of a walking leg were measured in different‐sized animals at 10 °C to compare the size scaling of oxygen consumption, whole‐body ETS activity and the ratio of whole‐body ETS activity to oxygen consumption (ETS/R). 2. Electron transport system activity of a leg and the ratio of ETS activity of a whole crayfish to that of a leg were correlated with wet mass of animals. Therefore, metabolic potential in whole noble crayfish can be estimated on the basis of the measured ETS activity in a single leg and crayfish mass. This approach provides a valuable tool for determining metabolic characteristics of crayfish without killing them. 3. Mass‐specific oxygen consumption decreased with increasing wet mass, while ETS activity of whole crayfish showed no significant correlation with wet mass. Both oxygen consumption and ETS activity correlated significantly with protein mass. 4. The increase in ETS/R with increasing wet mass of the noble crayfish indicates that small organisms exploit a greater proportion of their metabolic potential for standard metabolism than larger ones. This is the first report on ETS/R in crayfish.     

Size and structure of crayfish (Astacus astacus) populations on different habitats in Finland

The many inland waters in Finland make crayfish production an important potential resource. The rational utilization and management of this resource requires knowledge of the size and structure of the crayfish populations. The difficulties often encountered in catching crayfish complicate population studies. Mark-recapture and electric fishing have been used in the studies. The number of adult crayfish measuring more than 70 mm in a 4-ha lake was estimated at 620, and the number in a 13-ha lake at 3 480. In the lakes, the density of adult crayfish was around 0.6–1.4 m^–2 and in one stream studied about 2.5 m^–2 rising to several individuals per m² in the best biotopes.     

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.     

Structural and functional properties of the mechanoreceptors and chemoreceptors in the anterior oesophageal sensilla of the crayfish,Astacus astacus

Summary The anterior oesophageal sensilla (AOS) of Astacus astacus are short cuticular cylinders with a terminal pore. Three sensory cells belong to each AOS, two of them having a dendritic ciliary segment of about 2 m length (A-cells), the third having this segment about 12 m long (B-cell). The ciliary A-tubules in the B-cells only possess arms and an electron-dense core. The dendritic inner segments of the A-cells terminate 3–6 m distal of the B-cells. The dendritic tips of most A-cells are connected by desmosomes. All dendritic inner segments contain a ciliary rootlet and are accompanied by a scolopale within the innermost enveloping cell. There are four enveloping cells, three of which form thin subepithelial columns together with the enclosed sensory cells. Recordings from the posterior branch of the anterior oesophageal nerve containing the axons of the AOS revealed the presence of three types of sensory cells, two being chemosensitive and one mechanosensitive. One chemoreceptor is specifically sensitive to nicotinamide, but responded also to -NAD, 6-aminonicotinamide, nicotinamide methyl esther and nicotin. It was blocked by p(4)-acetylpyridine. The second chemoreceptor responded only to crayfish gastric fluid. The mechanoreceptors reacted to stretch of the oesophageal wall adapting only slowly to maintained stimuli. It is assumed that the A-cells are the chemosensitive cells and the B-cells the mechanosensitive ones. The latter show only a small number of modality-specific characteristics. Several structural features appear to be correlated with the location of the AOS within a flexible surface, which undergoes considerable dilation.Supported by the Deutsche Forschungsgemeinschaft (H.A.:SFB 4/G1; H.H.: HA 1201/1-4)     

The cytotoxic reaction of hemocytes from the freshwater crayfish, Astacus astacus

Crayfish hemocytes displayed cytotoxic capacity towards all tested mammalian tumor and nontumor cell lines. The ratio required for the cytotoxic action of effector cells to target cells was at least 1:1. The lysis of the target cells required a minimum of 1 hr to become detected. After separation and isolation of the hemocyte populations of crayfish, the semigranular and granular cells retained their cytotoxic capacity. These cells contain the prophenoloxidase activating (proPO) system, a complement-like pathway, which in an activated form lyses semigranular cells in vitro, but failed to kill the tested target cells.     

Comparative ultrastructure of the spermatozoa of three crayfish species: Austropotamobius torrentium,Pacifastacus leniusculus,and Astacus astacus (Decapoda: Astacidae)

This study reports about the spermatozoal ultrastructure of three species of astacid crayfish, i.e., the stone crayfish Austropotamobius torrentium, signal crayfish Pacifastacus leniusculus, and noble crayfish Astacus astacus. The acrosome is a cup shaped and electron‐dense structure at the anterior of the spermatozoon and comprises three layers of differing electron densities filled with parallel filaments that extend from the base to the apical zone. The acrosome was significantly longer in A. astacus than in P. leniusculus and the shortest acrosome belongs to A. torrentium. The width of the acrosome was significantly narrower in A. torrentium than in P. leniusculus and the widest acrosome belongs to A. astacus. The L:W ratio was significantly greater in A. torrentium than in P. leniusculus and the lowest ratio belongs to A. astacus. Radial arms are visible on each side of the acrosome or nucleus in sagittal view and wrap around the spermatozoon. Each radial arm comprises a parallel bundle of microtubules arranged along the long axis within a sheath. The nucleus, with decondensed material, is located in the posterior of the cell. All parts of the spermatozoon are tightly enclosed within an extracellular capsule. Despite a well‐conserved general structure and similarity of pattern among these spermatozoa, differences in the dimensions of the acrosome within the studied species may be useful to help distinguish the different crayfish species. J. Morphol., 2013. © 2013 Wiley Periodicals, Inc.     

Ontogeny of the antennal glands in the crayfish Astacus leptodactylus (Crustacea, Decapoda): immunolocalization of Na+,K+-ATPase

The involvement of the antennal urinary glands in the ontogeny of osmoregulatory functions was investigated during the development of Astacus leptodactylus by measurements of hemolymph and urine osmolality in juvenile and adult crayfish and by the immunodetection of the enzyme Na⁺,K⁺-ATPase. In stage II juveniles, 1-year-old juveniles, and adults, all of which were maintained in freshwater, urine was significantly hypotonic to hemolymph. In adults, chloride and sodium concentrations were much lower in urine than in hemolymph. During embryonic development, Na⁺,K⁺-ATPase was detected by immunocytochemistry in ionocytes lining the tubule and the bladder, at an eye index (EI) of 220–250 m, and in the labyrinth, at EI 350 m. In all regions, immunofluorescence was mainly located at the basolateral side of the cells. No immunofluorescence was detected at any stage in the coelomosac. In late embryonic stages (EI 410–440 m), in stage I juveniles, and in adults, strong positive immunofluorescence was found from the labyrinth up to and including the bladder. These results show that, as early as hatching, juvenile crayfish are able to produce dilute urine hypotonic to hemolymph. This ability originates from the presence of Na⁺,K⁺-ATPase in ion-transporting cells located in the labyrinth, the tubule, and the bladder of the antennal glands and constitutes one of the main adaptations of crayfish to freshwater.We thank 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 due to the Société Française dExportation des Ressources Educatives (SFERE) for the scholarship to S.K.     

Life-cycle and functional cytology of the hepatopancreatic cells of Astacus astacus (Crustacea,Decapoda)

Summary The hepatopancreas of the freshwater crayfish Astacus astacus was reinvestigated by means of light and electron microscopy using refined techniques of tissue preservation. The results contribute significantly to the solution of controversial problems of the decapod hepatopancreas such as cell genealogy, cellular interdependences, elimination of senescent cells and functional interpretation of the cell types. The three mature cell types of the organ, R-, F- and B-cells, are shown to originate independently from embryonic E-cells which are located at the blind-ending tips of the hepatopancreatic tubules. The less abundant M-cells are supposedly of non-hepatopancreatic origin since they are also found in other epithelia of the digestive tract. Differentiating cells can be assigned at an early stage to one of the three hepatopancreatic cell lines if the ultrastructural appearance and distribution pattern of their organelles are used as distinguishing features. The most sensitive markers are the Golgi bodies which have a cell-specific architecture and secretion product not only in mature cells but also in early differentiating stages. Later conversion of one cell type into another, as has often been proposed in literature, does not occur. Senescent cells are preferably expelled from the epithelium at the junction of neighbouring hepatopancreatic tubules and at the antechamber which links the hepatopancreas to the main digestive tract. Cellular discharge in the antechamber occurs by sliding of the oldest parts of the hepatopancreatic epithelium across a particular antechamber epithelium that was thus far unknown. New ultrastructural findings are described with respect to the absorptive apparatus of nutrient absorbing R-cells, the formation of Golgi vesicles and retrieval of membranes in digestive enzyme synthesizing F-cells, and the involvement of Golgi body and endoplasmic reticulum in the formation of heterophagic vacuoles in B-cells. The discovery of these ultrastructural features enables a more sophisticated functional interpretation of the hepatopancreatic cells of Decapoda.     

Patterns and projections of crustacean cardioactive-peptide-immunoreactice neurones of the terminal ganglion of crayfish

Three distinct clusters of crustacean cardioactive-peptide-immunoreactive neurones occur in the terminal abdominal ganglion of the crayfish species Orconectes limosus, Astacus leptodactylus, Astacus astacus and Procambarus clarkii, as revealed by immunocytochemistry of whole-mount preparations and sections. They exhibit similar topology and projection patterns in all four studied species. An anterior ventral lateral and a posterior lateral cluster contain one small, strongly stained perikaryon and two large, less intensely stained perikarya, each showing contralateral projections. A posterior medial lateral cluster of up to six cells also contains these two types of perikarya. Whereas the small type perikarya belong to putative interneurones, the large type perikarya give rise to extensive neurohaemal plexuses in perineural sheaths of the third roots of the fifth abdominal ganglia, the connectives, the dorsal telson nerves, the ganglion itself, its roots and arteriolar supply. Thin fibres from these plexuses reach newly discovered putative neurohaemal areas around the hindgut and anus via the intestinal and the anal nerves, and directly innervate the phasic telson musculature. A comparison with earlier investigations of motoneurones and segmentation indicates that these three cell groups containing putative neurosecretory neurones may be members of at least three neuromeres in this ganglion. Crustacean cardioactive peptide released from these neurones may participate in the neurohumoral and modulatory control of different neuronal and muscle targets, thereby exceeding its previously established hindgut and heart excitatory effects.Abbreviations AG abdominal ganglion - adpl arteria dorsalis pleica - Ala arreria lateralis abdominalis - Asub arteria subneuralis - CCAP crustacean cardioactive peptide - CNS central nervous system - IR immunoreactive - LG lateral giant axon - LTr lateral tract - MDT medial dorsal tract - MG medial giant axon - M Tr medial tract - mcan musculus compressor ani - mfltp museulus flexor telsonos posterior - nan nervus ani (AG6 N5) - nant nervus anterior (AG6 N1, N2) - nia nervus intestinal anterior - nin nervus intestinalis (AG6 N7) - nip nervus intestinalis posterior - nteld nervus telsonos dorsalis (AG6 N6) - nielv nervus telsonos ventralis (AG6 N4) - nur nervus uropedalis (AG6 N3) - nven nervus ventralis (AG5 N3) - PIR peri-intestinal ring - PTF posterior telson flexor - VLT ventral lateral tract - VMT ventral medial tract - VNC ventral nerve cord - VIF ventral telson flexor - AVLC, PLC, PMLC anterior ventral lateral, posterior lateral, posterior medial lateral CCAP-immunoreactive cell cluster - A6AVC, A7AVC anterior ventral commissures - A7DCI dorsal commissure I - A7PVC posterior ventral commissure - A7SCII sensory commissure II - A7VCII, A7VCIII ventral commissures II and III of the sixth (A6) and seventh (A7) abdominal neuromer     

The spermatogenesis of crustaceans. VI. The external morphology of the mature spermatozoa of mytilicola intestinalis,steuer (copepoda)

The external structure of the male gamete of Mytilicola intestinalis is studied under a scanning electron microscope and compared with transmission electron micrographs of thin sections corresponding to the different parts of same. The cell in question is long and filiform, showing, along a significant part of its length, two ridges or expansions helicoidally arranged and diametrically opposed. Four different parts or segments can be identified in this spermatozoon: segment A, characterized by its screw-like aspect; segment B, the longest, provided with well-developed helicoidal expansions; segment C, showing an uneven surface and lacking expansions; and segment D, with a smooth surface and decreasing diameter. The significance of this original structure in a spermatozoon, considered immobile until now, is discussed, stating different hypotheses with regard to the possible mobility of the cell just before fertilization takes place.     

The acute toxicity of fenitrothion on narrow-clawed crayfish (Astacus leptodactylus Eschscholtz, 1823) in association with biomarkers of lipid peroxidation

The aim of this research was to evaluate the acute toxicity of fenitrothion to the crayfish (Astacus leptodactylus Eschscholtz, 1823), which is chosen as an alternative aquatic organism to fish by using the static test system and evaluate the basic lipid peroxidation parameters for the first 24 h. Crayfish of 27.3 ± 0.56 g mean weight and 10.0 ± 0.72 cm mean length were selected for the bioassay experiments. The experiments were repeated three times in 20 liters of tap water. The temperature of water was 21 ± 1°C. The data obtained were statistically evaluated by using a computer program developed by the United States Environmental Protection Agency, based on Finney's probit analysis method and the 96-h LC(50) value for crayfish was calculated to be 15.75 μg/L. The 95% lower and upper confidence limits for the LC(50) were 9.45 to 25.01 μg/L. In addition to the acute toxicity bioassay experiments, 24-h oxidative stress parameters such as malondialdehyde (MDA) levels and ferrous oxidation assay (FOX HP [hydrogen peroxide] equivalents) were also determined. Only MDA levels of hepatopancreas decreased at 5, 10, and 20 μg/L of fenitrothion doses. We can conclude that fenitrothion is highly toxic to crayfish, a nontarget organism in the ecosystem, and the lipid peroxidation indicators can be easily used for monitoring environmental effects.     

Species-specific differences in dynamics of agonistic interactions may contribute to the competitive advantage of the invasive signal crayfish (Pacifastacus leniusculus) over the native narrow-clawed crayfish (Astacus leptodactylus)

Species-specific differences in dynamics of agonistic interactions may influence the outcome of interspecific competition and potentially contribute to competitive advantage of one species over another. In this study, we compared the dynamics of agonistic interactions of one of the most successful crayfish invaders of European freshwater ecosystems, the signal crayfish (Pacifastacus leniusculus) and the widespread native European species currently undergoing range expansion in Croatia and Europe, the narrow-clawed crayfish (Astacus leptodactylus). Comparisons between P. leniusculus pairs and the A. leptodactylus pairs demonstrated significant differences in frequency and duration of agonistic encounters: P. leniusculus dyads engaged in fewer fights, but their duration was significantly longer. In staged interactions between size-matched interspecific pairs, agonistic behaviour of P. leniusculus individuals translated into dominance over their A. leptodactylus counterparts. This indicates that the success of P. leniusculus in agonistic encounters with the native competitor might stem from its readiness to continue fighting for a longer time period, and could lead to ecological advantages during niche competition even when facing a successful native crayfish species.     

The morphology of the male reproductive system,spermatogenesis and the spermatozoon of Daphnia magna (Crustacea: Branchiopoda)

This study analyses the histological and cellular morphology of the testis and sperm development in the male Daphnia magna Straus 1820. Due to the rarity of males and predominately parthenogenetic lifecycle of Daphnia , there has been limited detailed information on males in contrast to the well‐studied female. Using light and electron microscopy approaches, we describe the morphology of the testis during the progression from an immature to mature testis. The testis has an encasing muscular mesh sheath outside the basal lamina, beneath which is a thin somatic epithelial cell layer. Internal to the epithelium are the spermatogonial stem cells and subsequent syncytial clusters of the germ cells as they progress through spermatogenesis; spermatozoa occupy the entire testis in sexually mature D. magna . We describe the structure of developing and mature spermatozoa; mature spermatozoa are non‐flagellated, ovoid in shape with plasmalemma filapodia and are encased in an extracellular capsule.