Morphofunctional study of the statocyst of the Cuban crayfish Procambarus cubensis

After replacing sand, a natural component of the endogenous otolithic apparatus of the Cuban crayfishProcamabrus cubensis by particles of reducible iron and subsequent exposure of the animals to a constant magnetic field, the following changes were noted in their behavior: during the first 24 h the motor activity of the animals increased and was accompanied by frequent strong beats of antennulae on water; animals tried to go away from the magnet attached to the aquarium wall and to hide in a shelter; by the day 10—12, there appeared signs of adaptation to the action of magnet; the animals spent most time near the wall with the magnet and were clinging close to it. If the magnet was moved the animals not only slipped to their side but also rotated around their longitudinal axis. The eyestalks also started moving. After molt and repeated introduction of reduced iron particles into the statocysts, a negative reaction to magnet was practically absent, and the crayfish preferred staying near the magnet. Study of structural organization of the statocysts under a scanning electron microscope showed that the iron particles in the otolithic apparatus of the crayfish are covered with a thinner layer of glycocalyx as compared to control animals whose otolithic apparatus is composed of sand grains. Besides, the otolithic apparatus of the experimental animals is a loose formation, unlike the dense glycocalyx rich in conglomerate in control animals.     

Evolution of Otolithic Membrane. Structure of Otolithic Membrane in Amphibians and Reptilians

Otolithic membrane of utricles, saccules, and lagena of amphibians (Bufo bufo, Xenopus laevis, Rana temporaria) and reptiles (Teratoscincus scincus, Agama sanguinolenta, Ophisaurus apodus, Caiman crocodilus) were studied using light and scanning electron microscopy. Otolithic membrane in various otolithic organs in all studied animals was found to differ by shape, size, structure, and composition of otoconia. Otolithic membrane of utricle of amphibians and reptiles represents a thin plate of non-uniform structure. Otolithic apparatus in saccule represents a large cobble-stone-like conglomerate of otoconia. Otolithic membrane of lagena looks like a bent plate and is poorly differentiated in amphibians, but well differentiated in reptiles. Thus, transition of vertebrates to the earth surface was accompanied by a fundamental reorganization of otolithic membrane structure. Otolithic membrane containing constantly growing large otolith (in fish) was replaced by a thin structurally differentiated otolithic membrane that ceases its growth at early stages of ontogenesis. However, this replacement did not occur simultaneously in all otolithic organs. The changes initially involved otolithic membrane of utricle. Saccule of amphibians and reptiles has a typical compositional otolith. In the course of further phylogenetic development of tetrapods the process of structural differentiation of otolithic membrane was enhanced and otoliths were completely lost. In parallel, there proceeded a process of replacement of prismatic and spindle-shaped aragonitic otoconia by calcitic barrel-shaped otoconia. The data obtained confirm our hypothesis put forward earlier about two directions of evolution of otolithic membrane.     

Crayfish as geomorphic agents and ecosystem engineers: effect of a biomass gradient on baseflow and flood-induced transport of gravel and sand in experimental streams

SUMMARY 1. Using experimental streams, we studied the impact of the crayfish Orconectes limosus on (i) the transport of gravel and sand at baseflow; (ii) the sediment surface (bedform, particle consolidation, proportion of sand, algal and gravel cover); and (iii) the critical shear stress (τ_C) causing incipient gravel and sand motion during simulated floods. We examined (i) and (ii) in experimental outdoor flumes that replicated riffle‐pool sequences and (iii) in a larger laboratory flume, in which we exposed sediments retrieved from the outdoor flumes to a progressively increasing discharge. 2. Habitat changes induced by crayfish, such as bedform alterations in riffles (downstream displacement of riffle heads) and the increase of gravel on sand dunes in pools, had major impacts on the spatial and temporal patterns of the baseflow transport of gravel and sand. 3. In addition to their impact on bedform in riffles and on gravel cover in pools, crayfish prevented the physical consolidation of particles in riffles and reduced the algal cover and the proportion of sand in the surface layer in both riffles and pools. These crayfish impacts on sediment surface variables had complex, interacting effects on the mobilisation of gravel and sand during subsequent flood simulations. For sand, crayfish progressively decreased the τ_C (i.e. the sum of bedform drag and skin friction) by about 50% along the entire biomass gradient in pools, whereas the presence of crayfish abruptly decreased the τ_C by about 75% in riffles. For gravel, the discharge causing motion in riffles produced a shear stress (in terms of skin friction) on an even bedform that was about 75% lower in all flumes with crayfish compared with the flumes without crayfish. Crayfish had no impact on τ_C for gravel in pools. 4. Scaling‐up these experimental results to real streams suggests that crayfish could affect the patch dynamics of major sediment transport events and habitat suitability for other organisms that, at larger spatial scales, could increase the overall spatio‐temporal habitat diversity and thus the overall structural and functional biodiversity of lotic communities.     

A Criterion for the Complete Deposition of Magnetic Beads on the Walls of Microchannels

This paper analyzes numerical simulations of the trajectories of magnetic beads in a microchannel, with a nearby permanent cubical magnet, under different flow and magnetic conditions. Analytically derived local fluid velocities and local magnetic forces have been used to track the particles. A centered position and a lateral position of the magnet above the microchannel are considered. The computed fractions of deposited particles on the walls are compared successfully with a new theoretically derived criterion that imposes a relation between the sizes of the magnet and the microchannel and the particle Stokes and Alfvén numbers to obtain the complete deposition of the flowing particles on the wall. In the cases in which all the particles, initially distributed uniformly across the section of the microchannel, are deposited on the walls, the simulations predict the accumulation of the major part of particles on the wall closest to the magnet and near the first half of the streamwise length of the magnet.     

The structure and growth of the statocyst in the Australian crayfish Cherax destructor

The morphology of the statocyst of the Australian crayfish Cherax destructor was examined using scanning electron microscopy. It resembles in general structure, size, and position the statocysts of crayfish described previously, and the size and distribution of the fields of setae on the floor of the capsule are similar but not the same. Over the size range examined, the relationship between the carapace length, the length of the basal antennular segment, the diameter of the statocyst capsule, and the total number of setae are all linear. The number and position of setae on the floor of the statocyst capsule were mapped for animals in two size classes (small, ca. 20 mm; large, ca. 50 mm) to test for changes in their arrangement during growth. The change in the ratio of setal number to statocyst size between the two size classes was about three times greater for the anterior setal field than for the other fields. We propose that differential development of the setal fields may be related to changes in the force-monitoring requirements of the animals as they increase in size, but this remains to be experimentally tested.     

Interspecific differences in crustacean homologous behavior: Neural mechanisms underlying the reversal of uropod steering movement

Summary Rolling of the body in the same direction induces an asymmetrical steering movement of uropods in the opposite directions in two closely related species of crayfish. InProcambarus clarkii the uropod on the upper side is spread out and closed on the lower side, whereas inCambaroides japonicus the uropod moves in the opposite direction. The stimulus detector, the statocysts, the effector, the uropod musculature, are neither structurally nor functionally significantly different in the two species. The results indicate that the opposite responses could be ascribed to differences in the interneuronal connections within the central nervous system of these two species.     

Modification of statocyst input to local interneurons by behavioral condition in the crayfish brain

Posture control by statocysts is affected by leg condition in decapod crustaceans. We investigated how, in the crayfish brain, the synaptic response of local interneurons to statocyst stimulation was affected by leg movements on and off a substratum. The magnetic field stimulation method permitted sustained stimulation of statocyst receptors by mimicking body rolling. The statocyst-driven local interneurons were classified into four morphological groups (Type-I–IV). All interneurons except Type-IV projected their dendritic branches to the parolfactory lobe of the deutocerebrum where statocyst afferents project directly. Type-I interneurons having somata in the ventral-paired lateral cluster responded invariably to statocyst stimulation regardless of the leg condition, whereas others having somata in the ventral-unpaired posterior cluster showed response enhancement or suppression, depending on the cell, during leg movements on a substratum, but no response change during free leg movements off the substratum. The synaptic responses of Type-II and IV interneurons were also affected differently by leg movements depending on the substratum condition, whereas those of Type-III remained unaffected. These findings suggest that the statocyst pathway in the crayfish brain is organized in parallel with local circuits that are affected by leg condition and those not affected.     

The diet of the spiny-cheek crayfish Orconectes limosus in the Czech Republic

The composition of the diet of the invasive spiny-cheek crayfish Orconectes limosus was studied using qualitative and quantitative analyses of stomach contents. A total of 368 specimens collected in 2003–2005 and 2008 in Czech localities were examined, predominantly from the Labe (Elbe) and Vltava River basins. Food components were compared for three size classes of crayfish and both sexes. The following conclusions were reached: (1) the spiny-cheek crayfish is an omnivorous species consuming plants, animals and detritus; (2) quantitatively, the main food component of O. limosus is detritus, while the plant component was second; (3) O. limosus may swallow whole food particles up to 4 mm in size, and the bodies of small animals may sometimes be found undamaged in their stomachs.     

Heteromorphic antennules protect the olfactory midbrain from atrophy following chronic antennular ablation in freshwater crayfish

Posthatch larval crayfish (Procambarus clarkii) were unilaterally antennulectomized and maintained in the laboratory for a 6-month period, during which time all regenerating antennular stumps were periodically excised. In another group of animals at a similar developmental stage a heteromorphic antennule was induced on the side of the head ipsilateral to the chronically sectioned normal antennule. After 6 months, all experimental animals were sacrificed and their brains were fixed and sectioned. Computer-aided quantitative measurements were obtained for the volumes of the olfactory lobes on both the experimental and control sides of the brains of both groups of crayfish. In the brains of the crayfish group in which only chronic antennualectomy had been performed, the olfactory lobe ipsilateral to the lesion was reduced in volume by about 80% compared to the olfactory lobe on the control side. In animals in which the normal antennule had been chronically ablated, but which possessed a heteromorphic antennule on the same side, the olfactory lobe on the lesioned side differed in volume from the control side by a mean value of only 28%. We conclude that afferent fibers from a heteromorphic antennule ipsilateral to a chronically lesioned normal antennule can assume some of the central trophic functions of the afferents from the normal antennule; thus, the presence of a heteromorphic antennule offers some measure of protection from the dystrophic effects of chronic ablation of the normal antennule during development and growth in crayfish.     

Fine Structure of a Silica-Biomineralizing Testate Amoeba,Netzelia tuberculata1

The fine structure of the cytoplasm and the intracytoplasmic origin of siliceous granules and surrounding cement plaques used in constructing the shell wall of Netzelia tuberculata are described. These organisms construct their test from biogenic siliceous particles and sand grains or other foreign particles (including starch grains apparently from algal prey) coated with biogenic silica. The smooth surface texture of the grains, compared to those of other particle-gathering testate amoebae, can be expalained by the deposition of a thin surface layer of silica on the foreign particles incorporated into the wall.     

Absorption of experimentally administered materials by the hepatopancreas cells of the crayfish, Procambarus clarki

After long term starvation, the crayfish, Procambarus clarki was administered protein silver, iron lactate and olive oil, and its hepatopancreas was subsequently examined by electron microscopy. The reserve cells showed changes suggesting the absorption of these materials from the acinar lumen had taken place. In contrast, the hindgut of crayfish seemed to have no absorptive ability. In crustaceans the hepatopancreas is the largest gland in the body. The chief functions of this gland are the secretion of digestive juice into the stomach and absorption of digested food. It is also where materials which are necessary for hardening of animals that have undergone ecdysis are stored. Although these roles are commonly accepted, the absorptive ability of the gland has been rarely studied. Yonge (1924) and van Weel (1955) attempted to obtain evidence for the absorptive function of hepatopancreas cells of Nephrops norvegicus and Atya spinides using iron lactate and iron saccharate, and obtained some positive results. They used the histochemical Prussian blue test to demonstrate absorbed iron. Vonk (1960) referred to the results of a few authors who had tried to show fat deposits in reserve cells of the hepatopancreas after the administration of olive oil to the animals. But because starvation did not affect the quantity of stored fat in the hepatopancreas cells, the attempt failed to reveal the absorption of fat by the hepatopancreas. In the present paper, the authors describe the results of studies on the absorption of experimentally administered materials by hepatopancreas cells of the crayfish, Procambarus clarki, using electron microscopy.     

Electromyographic analysis of the role of the otolith apparatus in regulation of postural disorders in humans with extrapyramidal pathology

In 30 patients with cervical dystonia in the posture of orthostasis, an obvious change of the electromyographic pattern was shown both at rest and in different functional tests. The analysis has shown an enhanced excitability of motoneurons and a disorder of the cortical control, as well as enhancement of the irradiation process between paired structures controlling activity of opposite muscular groups. After treatment with intratympanal administration of an ototoxical preparation on the opposite side in respect to head turn direction, normalising of the cortical control and diminishing of the excitation irradiation. The data obtained suggest a significant role of the vestibular otolithic apparatus in regulation of postural disorders in extrapyramidal pathology.     

A Test of Temporal Variation in Risk and Food Stimuli on Behavioral Tradeoffs in the Rusty Crayfish, Orconectes rusticus: Risk Allocation and Stimulus Degradation

The effects of temporal variation in exposure to predation risk on behavioral tradeoffs were tested in the rusty crayfish, Orconectes rusticus. Based on the risk allocation hypothesis, we predicted that increasing the frequency of encounter with predation risk would yield increasing responses to a food stimulus in the presence of both a risk stimulus and a food stimulus. Crayfish were exposed to risk every 12 h, every 6 h, or left undisturbed for 24 h prior to testing. The risk stimuli used were a plain water control, snapping turtle (Chelydra serpentina) cue, and conspecific alarm cue. After 24 h of conditioning, the crayfish were exposed to a combination of risk cue and food cue. The behavioral responses of the crayfish were recorded for 5 min immediately following the introduction of the cues and again for 5 min, 1 h after stimulus exposure. The crayfish were observed at the two times to determine how their responses to the combination of risk and food cues changed over time. The responses of the crayfish were significantly influenced by stimulus treatment, time, and the interaction of time and stimulus treatment. Further analysis indicated that responses to the stimulus treatments changed differently over time. Immediately after exposure, the crayfish were more active in the control and snapping turtle treatments than in the conspecific alarm treatment. The high levels of activity initially observed in the control and snapping turtle treatments waned over time, such that the behaviors recorded 1 h after exposure were not significantly affected by stimulus treatment. Neither frequency nor the interactions of frequency with stimulus and/or time significantly affected crayfish behavior. The results of this study did not support the risk allocation model and contrast with results from similar work on the virile crayfish, Orconectes virilis.     

Effects of fluoxetine on growth and behavior in the crayfish Orconectes rusticus

This laboratory study examined the effects of the specific serotonin reuptake inhibitor fluoxetine on growth following molting and on a range of behaviors in the crayfish Orconectes rusticus. For growth experiments, male Form I and Form II crayfish were weighed and measured and placed individually in water containing 0–500 μg/L of fluoxetine. They were held in fluoxetine or control water until they molted and were reweighed two weeks post-molt. In behavior experiments, juvenile and adult animals were held individually in 0, 2, 200, or 500 μg/L of fluoxetine for 10 days and tested in an open field arena to assess locomotion, thigmotaxis, sheltering, and habituation to a novel environment. Under our laboratory conditions, crayfish exposed to fluoxetine at 500 μg/L showed significantly enhanced growth: post-molt Form I animals had greater body weight and post-molt Form II animals had greater carapace length, relative to controls. In open field tests, juvenile crayfish exposed to 2 and 500 μg/L fluoxetine displayed significantly reduced locomotion compared to controls. The results indicate that crayfish growth and locomotion can be manipulated by short-term exposure to ambient fluoxetine, suggesting that this means of exposure may offer a useful and noninvasive way to examine drug effects in freely moving animals. However, effects were only observed at concentrations well above fluoxetine levels currently reported in the environment. This suggests that O. rusticus may be relatively resistant to this form of pharmaceutical pollution but whether effects would occur following long-term exposure to lower concentrations is unknown.     

“Spinner” cephalopods: defects of statocyst suprastructures in an invertebrate analogue of the vestibular apparatus

Summary Individuals of seven species of coleoid cephalopods (three species of octopus, three of squid, and one of cuttlefish), that were cultured and reared in laboratory aquarium systems, had a behavioral defect at hatching which was characterized by an inability to control orientation while swimming. These defective animals were designated as spinners.An examination of statocysts from individuals of five of the affected species revealed abnormalities of the neuroepithelial suprastructures: absence or malformation of the statolith of the gravity receptor system and absence of the cupulae of the angular acceleration receptor systems. The sensory epithelia did not differ from those of normal animals, nor did the synaptic structures and relationships, when examined both with scanning and transmission electron microscopy. The abnormalities were compared with congenital defects of the neuropeithelial suprastructures of the vestibular apparatus (especially in mammals). The defects observed in statocysts of spinner animals are thought to be the result of environmental causes, such as the temperature or chemistry of the seawater in the transportation vessels or rearing systems, rather than genetic causes.     

Niche differentiation among invasive crayfish and their impacts on ecosystem structure and functioning

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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.     

Transport of rosettes from the golgi apparatus to the plasma membrane in isolated mesophyll cells ofZinnia elegans during differentiation to tracheary elements in suspension culture

Summary Rosettes of six particles have been visualized by freeze-fracture in the protoplasmic fracture (PF) faces of: a) the plasma membrane, b) Golgi cisternae, and c) Golgi-derived vesicles in mesophyll cells ofZinnia elegans that had been induced to differentiate synchronously into tracheary elements in suspension culture. These rosettes have been observed previously in the PF face of the plasma membranes of a variety of cellulose-synthesizing cells and are thought to be important in cellulose synthesis. InZinnia tracheary elements, the rosettes are localized in the membrane over regions of secondary wall thickening and are absent between thickenings. The observation of rosettes in the Golgi cisternae and vesicles suggests that the Golgi apparatus is responsible for the selective transport and exocytosis of rosettes in higher plants, as has been previously indicated in the algaMicrasterias (Giddings et al. 1980). The data presented indicate that the Golgi apparatus has a critical role in the control of cell wall deposition because it is involved not only in the synthesis and export of matrix components but also in the export of an important component of the cellulose synthesizing apparatus. The rosettes are present in the plasma membrane and Golgi vesicles throughout the enlargement of the secondary thickening, suggesting that new rosettes must be continually inserted into the membrane to achieve complete cell wall thickening.Abbreviations EF Golgi vesicles, exoplasmic fracture; the plasma membrane, extracellular fracture - PF protoplasmic fracture     

Sand accumulation in the digestive tract of rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus): The role of the appendix

We determined location and amount of accumulated sand in the gastrointestinal tract (GIT) of rabbits (Oryctolagus cuniculus) and guinea pigs (Cavia porcellus) fed diets containing external (silicate) abrasives. Computed tomographic abdominal images of rabbits (n = 44) and guinea pigs (n = 16) that each received varying numbers (4–7) of different diets for 14 days each (total n = 311 computed tomographs), and radiographs of dissected GIT and presence of silica in GIT content (n = 46 animals) were evaluated. In rabbits, the majority of accumulated sand was located in the caecal appendix, an elongated, intestinal structure in the left side of the abdomen. The ‘wash-back’ colonic separation mechanism in rabbits may be partly responsible for a retrograde transport of sand back to the caecum, where dense, small particles accumulate in the appendix. The appendix likely acted as a reservoir of these particles, leading to significant effects not only of the momentary but also of the previous diet on recorded sand volumes in the rabbits. Guinea pigs have no caecal appendix and a colonic separation mechanism not based on a ‘wash-back’. Less sand accumulation was found in their GIT without a specific location pattern, and there were less previous diet effects in this species. None of the rabbits or guinea pigs developed clinical signs of obstruction during the study, and the recorded sand volumes represented 1.0 ± 1.2% of the 14-d sand intake in rabbits and 0.2 ± 0.2% in guinea pigs. Accumulation of sand in volumes up to 10 cm³ in the GIT of rabbits does not seem to cause clinical health impairment. Large inter-individual differences in rabbits indicate inter-individual variation in proneness to sand accumulation. The reason for the presence of a sand-trapping caecal appendix in animals that are, due to their burrowing lifestyle and feeding close to the ground, predestined for accidental sand ingestion, remains to be unveiled.     

Morphological convergence in a Mexican garter snake associated with the ingestion of a novel prey

Morphological convergence is expected when organisms which differ in phenotype experience similar functional demands, which lead to similar associations between resource utilization and performance. To consume prey with hard exoskeletons, snakes require either specialized head morphology, or to deal with them when they are vulnerable, for example, during molting. Such attributes may in turn reduce the efficiency with which they prey on soft‐bodied, slippery animals such as fish. Snakes which consume a range of prey may present intermediate morphology, such as that of Thamnophiine (Natricinae), which may be classified morphometrically across the soft–hard prey dietary boundary. In this study, we compared the dentition and head structure of populations of Thamnophis melanogaster that have entered the arthropod–crustacean (crayfish)‐eating niche and those that have not, and tested for convergence between the former and two distantly related crayfish specialists of the genus Regina (R. septemvittata and R. grahamii). As a control, we included the congener T. eques. Multivariate analysis of jaw length, head length, head width, and number of maxillary teeth yielded three significant canonical variables that together explained 98.8% of the variance in the size‐corrected morphological data. The first canonical variable significantly discriminated between the three species. The results show that head dimensions and number of teeth of the two Regina species are more similar to those of crayfish‐eating T. melanogaster than to non‐crayfish‐eating snakes or of T. eques. It is unclear how particular head proportions or teeth number facilitates capture of crayfish, but our results and the rarity of soft crayfish ingestion by T. melanogaster may reflect the novelty of this niche expansion, and are consistent with the hypothesis that some populations of T. melanogaster have converged in their head morphology with the two soft crayfish‐eating Regina species, although we cannot rule out the possibility of a morphological pre‐adaptation to ingest crayfish.