Photoperiod affects gamete production, and protein and lipid metabolism in male narrow-clawed Crayfish Pontastacus leptodactylus (Eschscholtz, 1823).

Understanding effects of photoperiod on the reproductive efficiency and physiological responses of Pontastacus leptodactylus is important for successful management of the crayfish hatcheries. Spermatozoal number, gonado-somatic index (GSI), hepatosomatic index (HSI), testicular index (TI), vas deferens index (VDI), and total protein, amino and fatty acid profiles of the hepatopancreas were evaluated in male narrow-clawed crayfish P. leptodactylus. Animals (30-70 g) were maintained utilizing five different photoperiodic regimens (3 L:21D, 6L:18D, 24 L:0D, 0L:24D, and natural photoperiod (9 L:15D)) for 5 weeks. Different photoperiods affected spermatozoal number, GSI, and total protein, amino acid and fatty acid profiles of the hepatopancreas (P < 0.05). The greatest spermatozoal number and GSI were recorded in darkness (P < 0.05). There was the least spermatozoal number (3.5 × 106 ± 0.5) and GSI (2 ± 0.1%) when imposing the 9 L and 24 L photoperiodic regimens, respectively. In addition, maintenance in constant light as compared with other lighting regimens resulted in a greater total protein (43.68 ± 4.83 mg/g), phenylalanine (7.23 ± 0.35%), and total single unsaturated fatty acids (ΣMUFA) (48.07 ± 2.30%), but lesser total polyunsaturated fatty acids (ΣPUFA) (23.36 ± 1.07%), serine (1.53 ± 0.02%), Σn-6 (11.24 ± 1.18%), 20:4 (n-6) (2.7 ± 0.14%), and 20:2 (0.67 ± 0.17%) contents in the hepatopancreas (P < 0.05). The results of the present study indicate total darkness increases the spermatozoal production and imposing longer dark periods improved the reproductive efficiency and physical conditions in male P. leptodactylus.

Ultrasound treatment of frozen crayfish with chitosan Nano-composite water-retaining agent: Influence on cryopreservation and storage qualities.

The synergistic impact of ultrasound and Nano-water retaining agent on the cryoprotective effect on crayfish during frozen storage was investigated. The samples soaked in water-retaining agent (WRA), ultrasonic 60 W treatment combined with water-retaining agent (WRA-US60W) and ultrasonic 80 W treatment combined with water-retaining agent (WRA-US80W) were frozen storage at -18 °C. The indices of frozen storage in 0, 7, 14, 21, 28, 35 and 42 days were measured. The results showed that there was a significant difference in the soaking weight gain, thawing loss, water content and water activity between control group and WRA groups (P < .05). The ultrasound combined with WRA treatment showed better water retention effect. The water holding capacity of heat induced gel decreased continuously during the frozen storage period, and the WRA-US60W group exhibited significantly higher values than that of other treatment groups (P < .05). The texture characteristics of hardness, elasticity and chewiness had a significant change (P < .05). The development of total volatile base nitrogen content, myofibrillar protein content and Ca2+-ATPase activity of muscle protein were significantly delayed by WRA-US60W treatment, maintaining the integrity of tissue structure. Therefore, WRA-US60W treatment was found effectively improving the quality of crayfish during frozen storage.

Do arthropods feel anxious during molts?

The molting process of arthropods, chiefly controlled by ecdysteroids, is generally considered very stressful. Our previous investigations have shown that crayfish, after having experienced stressful situations, display anxiety-like behavior (ALB), characterized by aversion to light in a dark/light plus-maze (DLPM). In the present experiments, the spontaneous exploratory behavior of isolated crayfish was analyzed in a DLPM at different stages of their molt cycle. All tested animals displayed transitory aversion to light similar to ALB, before and, mostly, after molting, but not during inter-molt. Injection of ecdysteroids into inter-molt animals elicited ALB after a delay of 4 days, suggesting a long-term, possibly indirect, hormonal effect. Importantly, ecdysteroid-induced ALB was suppressed by the injection of an anxiolytic benzodiazepine. Thus, molts and their hormonal control impose internal stress on crayfish, leading to aversion behavior that has the main characteristics of anxiety. These observations are possibly generalizable to many other arthropods.

Left-right asymmetry in firing rate of extra-retinal photosensitive neurons in the crayfish.

The purpose of this paper is to explore the firing rate of the caudal photoreceptors (CPRs) from the sixth abdominal ganglion of the crayfish Cherax quadricarinatus. We use simultaneous extracellular recordings on left and right CPR in the isolated ganglion (n = 10). The CPRs showed an asymmetry in the spontaneous activity and light-induced response. In darkness, we observed one subgroup (70%) in which the left CPR (CPR-L) and right CPR (CPR-R) had spontaneous firing rates with a median of 18 impulses/s and 6 impulses/s, respectively. In another subgroup (20%), the CPR-R had a median of 15 impulses/s and the CPR-L had 8 impulses/s. In both groups, the differences were significant. Furthermore, the CPRs showed an asymmetrical photoresponse induced by a pulse of white light (700 Lux, 4 s). In one subgroup (30%), the CPR-L showed light-induced activity with a median of 73%, (interquartile range, IQR = 51), while the CPR-R had a median of 41%, (IQR = 47). In another subgroup (70%), the CPR-R showed a median of 56%, (IQR = 51) and the CPR-L had a median of 42%, (IQR = 46). In both groups, the differences were significant. Moreover, we observed a differential effect of temperature on CPR activity. These results suggest a functional asymmetry in both activities from left and right CPRs. These CPR activity fluctuations may modulate the processing of information by the nervous system.

Managing of Procambarus clarkii by X-ray sterilisation of males: Cytological damage to gonads.

Procambarus clarkii is an invasive alien species spreading worldwide. It is therefore mandatory to find new methods to manage this species since traditional techniques are not sufficient for this purpose. The present study investigates gonad damage induced by different doses of ionising irradiation: 20, 40 and 60 Gy. Testis were analysed after 10 and 30 days by means of light, scanning and transmission electron microscopy. Control unirradiated testes present an acinar structure with a well-defined germinative cells maturation from the distal proliferative zone to the proximal stalk of the lobes whilst, in irradiated testes, induced apoptosis of germinative and accessory cells and a high level of vacuolisation inside the acini were identified, progressively increasing in accordance to Gy dosage and time after exposure. We determined the dose of 40 Gy as the best compromise: it causes an extensive damage to germinative tissues without affecting crayfish vitality, differing from 60 Gy. From an applicative point of view, this dose reduces the efforts, in terms of cost and time, for the application of SMRT.

Effects of uranium on crayfish Procambarus clarkii mitochondria and antioxidants responses after chronic exposure: what have we learned?

We examined the impacts of Uranium (U) on mitochondria and on the response of antioxidants in the gills and the hepatopancreas of crayfish Procambarus clarkii after long-term exposure (30 and 60 days) to an environmentally relevant concentration (30 µg U/L). The expression of mitochondrial genes (12s, atp6, and cox1), as well as the genes involved in oxidative stress responses (sod(Mn) and mt) were evaluated. The activities of antioxidant enzymes (SOD, CAT, GPX and GST) were also studied. U accumulation in organs induced changes in genes' expression. The evolution of these transcriptional responses and differences between gene expression levels at high and low doses of exposure were also discussed. This study demonstrated that, after long-term exposure, U caused a decrease in antioxidant activities and induced oxidative stress. A possible ROS-mediated U cytotoxic mechanism is proposed. Expression levels of the investigated genes can possibly be used as a tool to evaluate U toxicity and seem to be more sensitive than the enzymatic activities. However a multiple biomarker approach is recommended as the perturbed pathways and the mode of action of this pollutant are not completely understood.

Effects of short time UV-A exposures on compound eyes and haematological parameters in Procambarus clarkii (Girad, 1852).

The amount of ultraviolet radiation (UVR) reaching the Earth's surface has been increasing as a result of an increasingly thinner ozone layer. The UV-A component of the UVR is able to generate oxidative stress in the compound eye and haemolymph of Procambarus clarkii when the latter was exposed for as little as 15 min daily for one week to UV-A. Changes in the eye involved corneal material, crystalline cones, pigments in cone stalks and retinula cells, rhabdom integrity, haemocyte infiltration, and haemal spaces. UV-A had significant impacts on haemolymph iron and glucose, whereas Ca ions were unaffected. Total protein and Cu-ions showed only insignificant changes following UV-A radiation. Involvement of lipid peroxidation and DNA fragmentation was significant with regard to the tissue damage cause by the UV-A. UV-A furthermore induced biological effects on serum electrophoretic patterns: some fractions either increased in size or others decreased. The described changes can be used as reference guidelines in evaluations of UV-A induced stress effects in P. clarkii.

[Ecological and radio-ecological effects from long-term use of the lake Kyzyl-Tash as a cooling reservoir by the nuclear fuel cycle facility].

This review introduces long-term study findings on ecological and radiation induced regime of the water reservoir - lake Kyzyl-Tash (R-2) - used as a heat sink of nuclear-power reactors in the Southern Urals from 1948 through 2008. It was exhibited that water reservoir exploitation by the nuclear fuel cycle facility "Mayak" PA resulted in hydrological, thermal, hydrochemical and radiological ecosystem regimes changes. The central radioactive substances depot in the water reservoir was determined to be the upper 20-30 cm bed silt layers, contamination density of which in 1980-1990s amounted on average approximately 0.2 PBq/km2 (about 5 kKu/km2). Some regularities of radionuclide distribution in bed sediments and biota were ascertained. Dose estimates from ionizing exposure to fish inhabited the water reservoir were experimentally made. Dose contribution was mainly due to incorporated beta-emitters amounted up to 2-3 Gy/y in 1980s. The leading role in the reservoir life belonged to phytoplankton with its algal nuisance periodicity constituting 5-6 years for blue-green and diatomic algae, and 2-3 years for green algae. During periods of the highest development pressure phytoplankton productive capacity in the reservoir was by an order of magnitude greater compared to control water reservoirs of the region. Combined long-term impact of radiological and chemical factors did not cause irreversible changes either in fish populations or ecological system in general. It can be proved by the fact that during 1970-1980s the water reservoir R-2 was inhabited by such cleanness indicators as crawfish (Astacus leptodactylus) and shellfish (Anodonta cygnea L.). On reducing of thermal and chemical pressure in the end of 1980s some processes observed gave evidence of ecosystem restoration in the lake Kyzyl-Tash. At the present moment the situation of the water reservoir exploiting as a heat sink is stabilized with preserved self-cleaning capacity.

Effect of pigment dispersing hormone on the electrical activity of crayfish visual photoreceptors during the 24-h cycle.

Visual photoreceptors are structures involved in the expression and synchronization of crayfish circadian rhythm of sensitivity to light (electroretinogram, ERG). Considering the relevant role of Pigment dispersing hormone (PDH) in the invertebrate circadian system organization, we study the effect of this substance on the electrical activity of crayfish visual photoreceptors during the 24-h cycle. The study demonstrates that: (1) PDH affects the electrical response to light of crayfish visual photoreceptor cells in a circadian time-dependent manner. (2) The kinetics of the light-elicited current of crayfish visual photoreceptor cells, as well as the ionic permeability underlying the electrical response to light vary over the 24-h cycle. (3) PDH modifies the kinetics and ionic permeability underlying the light-elicited current of crayfish visual photoreceptor cells in a circadian time-dependent manner.

Melatonin modulates the ERG circadian rhythm in crayfish.

One of the most important functions modulated by melatonin is the synchronization of circadian rhythms. In crayfish (Procambarus clarkii), we have obtained evidence that the amplitude of the electrical response to light of the retinal photoreceptors the receptor potential, is modified by the action of melatonin and that the magnitude of this action depends on the circadian time of melatonin application. In contrast, the electroretinogram (ERG) circadian rhythm can be synchronized by either single or periodic melatonin application. In this work we hypothesized that, in crayfish, melatonin acts on effectors and on pacemaker of ERG circadian rhythm as a non-photic synchronizer. Melatonin could be a hormone that sends a signal of darkness to the ERG circadian system.

Pigment dispersing hormone generates a circadian response to light in the crayfish, Procambarus clarkii.

Photoreceptor cells have been identified as important structures in the organization of the circadian system responsible for the generation and expression of the electroretinogram (ERG) circadian rhythm. They are the structures where the circadian periodicity is expressed (effectors) and which transform information from external light signals to be conducted to the pacemaker in order to induce adjustments of the rhythm (synchronizers). After isolation, eyestalks perfused in a pigment dispersing hormone (PDH) solution, show significant changes in receptor potential (RP) amplitude and duration. Exogenous PDH injected into intact crayfish induces a migration of retinal shielding pigments to a light-adapted state. A single dose of PDH produces advances or delays in the circadian rhythm of response to light of visual photoreceptors. All these effects depend on the circadian phase of PDH application. Consequently, the determination of the action of exogenous PDH on photoreceptor cells proved to be very helpful in understanding some mechanisms underlying the circadian organization of crayfish.

Spontaneous and light-evoked discharge of the isolated abdominal nerve cord of crayfish in vitro reveals circadian oscillations.

We examined the well-known spontaneous discharge (SD) and light-evoked (PD) discharge of the crayfish caudal photoreceptor for the possible existence of a daily rhythm in spike frequency. To do this, we isolated the abdominal nerve cord in vitro and studied its discharge frequency in constant darkness. Single cosinor analysis revealed significant SD and PD circadian rhythms (P < .05) with periods tau = 24.4h and 24.2h, respectively. These oscillations correspond to an endogenous circadian discharge of the caudal photoreceptor that is enhanced by light. The importance of this rhythm in the adaptive behavior of crayfish is discussed.

Millimeter-wave effects on electric activity of crayfish stretch receptors.

The effects of super high frequency (SHF) microwaves (34-78 GHz) on rates of spontaneous firing of the slowly adapting, stretch-receptor neurons of crayfish were studied. Initially, irradiation of continuously perfused, fluid-cooled preparations at power densities to 250 mW/cm2 caused a transient decrease in the rate of spontaneous firing (the dynamic response). Subsequently, with extinction of the SHF field, the rate of firing increased, finally stabilizing at pre-exposure levels (stationary phase). Rates of firing also increased when the receptor muscle was stretched, and they were inversely correlated with small, field-induced increases of temperature (approximately 1.5 degrees C). The response to SHF radiation did not depend on frequency if temperature of the medium was constant. No resonant peaks were found when the millimeter range of frequencies was scanned.

[Effects of artificial light, distributed through long photophases, on the annual growth cycle of the crayfish Orconectes limosus (Raffinesque)]. / Effets de la lumière artificielle, dispensée en longues photophases, sur le cycle annuel de croissance de l'ecrevisse Orconectes limosus (Rafinesque)

A photophase of 18 hrs interrupts the annual anecdysis of Orconectes limosus, which does not constitute an irrefutable evidence of a photoperiodic control in the cessation of anecdysis, since molts are also induced during the annual rest by an abnormally high temperature after a stay at low temperature. On the contrary, long photophases (18 and 24 hrs), dispensed from the beginning of the molting period, do not inhibit the start of the anecdysis of adults on the express condition that the covers are reconstituted.