Novelty stress and reproductive state alters responsiveness to sensory stimuli and 5-HT neuromodulation in crayfish

Sensory stimuli can produce varied responses depending on the physiological state of an animal. Stressors and reproductive stage can result in altered biochemical status that changes the responsiveness of an animal to hormones and neuromodulators, which affects whole animal behavior in relation to sensory stimuli. Crayfish serve as a model for examining the effects of neuromodulators at the neuromuscular junctions (NMJs) and for alterations in stereotypic behaviors for particular stimuli. Thus, we used crayfish to examine the effect of novelty stressors in males and the effect of being gravid in female crayfish to exogenous application of serotonin (5-HT). The responsiveness of neuromuscular junctions to 5-HT revealed that stressed as well as gravid crayfish have a reduced response to 5-HT at NMJs. The stressed crayfish were not fatigued since the basal synaptic responses are large and still showed a pronounced response to 5-HT. Using intact animals to examine a tail flip behavior, we showed that the rate of habituation in tail flipping to a strong repetitive stimulus on the telson is reduced in stressed males. Gravid females show no tail flipping behavior upon telson stimulation.     

Effects of an introduced crayfish, Procambarus clarkii, on the distribution of south-western Iberian amphibians in their breeding habitats

The introduction of exotic species in aquatic habitats is one of the causes for the amphibian declines observed worldwide. In the 1970s, the red swamp crayfish Procambarus clarkii was introduced in the southwest Iberian Peninsula, where no native crayfish occur. In this study we assess the effect of P. clarkii presence in the breeding site distribution of each of the 13 southwest Iberian amphibians, while simultaneously accounting for the effects of potentially confounding habitat variables, as well as for the effects of the other large aquatic predators in the study area – predatory fish. Amphibian species richness was lower in places where P. clarkii was present than in places without P. clarkii , regardless of fish presence. After accounting for habitat variables and fish, crayfish presence was a negative predictor of the breeding probability for all urodeles ( Pleurodeles waltl , Salamandra salamandra , Triturus boscai and T. marmoratus ) and for two anurans ( Pelobates cultripes and Bufo bufo ). The majority of the species affected usually breed in temporary ponds without fish, but that may be colonized by the crayfish. The ongoing expansion of P. clarkii may eventually lead to a growing isolation of amphibian populations and ultimately to local extinctions and a permanent alteration of the amphibian communities in southwest Iberian Peninsula.     

Effect of variation in photoperiod and light intensity on oxygen consumption, lactate concentration and behavior in crayfish Procambarus clarkii and Procambarus digueti

The effects of light intensity and duration on metabolic and behavioral parameters of two species of crayfish, Procambarus clarkii and Procambarus digueti, were studied. Sixty animals of each species were submitted to high irradiance conditions of two different photoperiod lengths, one normal light/dark (LD) 12:12 and one extreme LD 20:4 for 2 weeks. Hemolymph, lactate and oxygen consumption were determined throughout the experimental period. Simultaneously in 18 additional animals of each species, motor activity was individually recorded under the same control and experimental conditions. Both species showed a decrease in oxygen uptake and an increase in hemolymph lactate concentration. The statistical significance of this finding was higher for LD 20:4. This extreme condition evoked a significant decrease of motor activity in P. clarkii and a high mortality rate in P. digueti. P. digueti did not survive after the experiment, whereas P. clarkii survived and adapted to the laboratory conditions. Changes in metabolic and behavioral parameters could indicate different adaptation abilities in these species.     

Mutual mate choice in crayfish: large body size is selected by both sexes, virginity by males only

Mate choice is often assumed to be a prerogative of females because of their putatively larger reproductive investment than males. However, recent evidence suggests that spermatogenesis is far from being limitless and that males show a high selectivity towards their mates, thus maximizing their reproductive success. We investigated mutual mate choice in the crayfish Procambarus clarkii through two experiments. The first experiment explored the effects of body size, chelar size and chelar symmetry and social status of a potential partner. In the second experiment, we asked whether this species can discriminate between partners of the same body size but with different mating status. We used a binary choice test paradigm, in which two 'targets' with opposing characteristics were simultaneously presented to a test animal, the 'chooser'. The results showed that P. clarkii males are more selective than expected. Similar to the other sex, they were significantly attracted by targets with large body sizes, but not by individuals with larger and symmetric chelae or with a dominant status. An inter-sexual difference was found in the second experiment, in which only males seemed to select virgin potential mates. The several adaptive explanations for these preferences, still under debate, are finally discussed.     

Physiological biomarkers of hypoxic stress in red swamp crayfish Procambarus clarkii from field and laboratory experiments

The crayfish industry in Louisiana is the largest in the United States, with crayfish frequently harvested from waters that experience episodic or chronic hypoxia (dissolved oxygen [DO]≤ 2 mg/l). We examined physiological biomarkers (hemolymph lactate, glucose, and protein concentrations) of hypoxic stress in the red swamp crayfish Procambarus clarkii from chronically hypoxic natural habitats and laboratory hypoxia experiments. P. clarkii from normoxic and hypoxic areas in the Atchafalaya River Basin were sampled monthly from April to July 2010. Laboratory experiments subjected P. clarkii to severe hypoxia (1 mg/l DO), moderate hypoxia (2 mg/l DO), or normoxic conditions (control: DO>7.5 mg/l) for 12, 24, and 48 h. P. clarkii from normoxic and hypoxic natural habitats did not display significantly different hemolymph lactate or glucose concentrations; however, mean hemolymph protein concentration was significantly lower in crayfish from hypoxic areas. P. clarkii exposed to severe hypoxia in laboratory experiments had significantly higher hemolymph lactate and glucose concentrations for all three exposure times, whereas large differences in protein concentrations were not observed. These results suggest that elevated hemolymph lactate and glucose concentrations are responses to acute hypoxia in P. clarkii, while differences in protein concentrations are the result of chronic hypoxic exposure.     

Behavioral effects of serotonin and serotonin agonists in two crayfish species, Procambarus clarkii and Orconectes rusticus

Exogenous serotonin elicits several behaviors in Procambarus clarkii, including a flexed, elevated posture, reduced locomotion, and changes in aggressive behavior. We conducted experiments to determine if several serotonin agonists mimicked the behavioral effects of serotonin in two crayfish species, P. clarkii and Orconectes rusticus. Drugs tested were 1-(3-Chlorophenyl)-piperazine dihydrochloride (mCPP), Oxymetazoline, 2,5-dimethoxy-4-iodoamphetamine (DOI), CGS-12066A, and (+/-)-8-hydroxy-2-(di-n-dipropylamino) tetralin (8-OH-DPAT). In P. clarkii, mCPP most closely mimicked the effects of serotonin, significantly increasing the performance of the flexed, elevated posture and reducing locomotion; 8-OH-DPAT significantly reduced locomotion as well. Both of these drugs produced significant increases in elevated posture and decreases in locomotion in O. rusticus, and in this species, the drugs at test concentrations were more effective in eliciting these effects than serotonin. The effects of the drugs on behaviors performed during fighting bouts were variable. In both species, only 8-OH-DPAT significantly reduced several agonistic behaviors, and no agonist or 5-HT itself produced significant increases in agonistic behavior.     

克氏原螯虾幼体及亚成体对中华鳖的趋避行为

【背景】在我国,克氏原螯虾是一种外来入侵水生生物,已经对入侵地的生态环境造成严重的破坏。【方法】在Y形水迷宫中,研究有经验和无经验的幼体及亚成体克氏原螯虾对中华鳖的趋避行为。【结果】幼虾无论是否有经验都会显著回避中华鳖,降低外倾性;而亚成体虾对中华鳖的反应不及幼虾灵敏。【结论与意义】2种不同发育阶段的外来入侵物种克氏原螯虾均会回避土著物种中华鳖,为外来入侵生物防治和动物行为学及相关研究提供了参考。     

Grazing by a native and an exotic crayfish on aquatic macrophytes

1. We compared grazing by native noble crayfish ( Astacus astacus ) and the exotic signal crayfish ( Pacifastacus leniusculus ) on seedling or well-established macrophytes.
2. In a pool experiment, seedlings of emergent Scirpus lacustris and floating-leaved Potamogeton natans were heavily grazed by adult signal crayfish, whereas established plants of the same species sustained only minor damage.
3. In a preference experiment two submerged macrophytes ( Chara vulgaris and Elodea canadensis ), and both seedlings and established plants of S. lacustris and P. natans , were presented pairwise to signal and noble crayfish. There was no significant difference in preference by the two crayfish species. Chara vulgaris was preferred to all other plants presented, established plants of S. lacustris and P. natans were never preferred, and seedlings of P. natans were preferred to established P. natans .
4. An aquarium experiment was conducted in which the consumption of Chara by signal and noble crayfish was measured in relation to water temperature. Signal crayfish consumed significantly more Chara than noble crayfish, especially at higher temperatures.
5. Our results indicate that the signal crayfish is the more voracious grazer especially at higher temperatures. There may be negative effects on vegetation (emergents and floating-leaved, as well as submerged species) when the signal crayfish is introduced. Chara species are particularly susceptible, since they are preferred by crayfish and the genus includes a large number of rare species. Stocking of crayfish therefore could lead to the decline or removal of submerged species in order of crayfish feeding preferences and could prevent the spread or cause a decline of emergent and floating-leaved vegetation.     

A Comparison of Trace Metal Accumulation in Indigenous and Alien Freshwater Macro-Decapods

Concentrations of Cd, Cu, Fe, Mg, Mn, Ni, Pb and Zn were determined in the exoskeleton and hepatopancreas of three freshwater macro-decapods widely distributed in Tuscany, Italy. The species were the indigenous Austropotamobius pallipes (the white-clawed crayfish) and Potamon fluviatile (the river crab) and the alien Procambarus clarkii (the red swamp crayfish, in Italy aquacultured since 1977 and invasive since 1989). Water and sediments from the three environments in which they occur were analysed for trace metals to evaluate the degree of contamination of the habitats. Procambarus clarkii showed a significantly higher accumulation of most of the metals analysed, including the non-essential cadmium, nickel and lead. Interspecific differences were not related to exposure to higher concentrations of metals, but indigenous and alien macro-decapods possibly differ in their physiological requirements for essential trace metals and/or in their rates of essential and non-essential metal uptake, accumulation, detoxification and excretion.     

A Comparison of Trace Metal Accumulation in Indigenous and Alien Freshwater Macro-Decapods

Concentrations of Cd, Cu, Fe, Mg, Mn, Ni, Pb and Zn were determined in the exoskeleton and hepatopancreas of three freshwater macro-decapods widely distributed in Tuscany, Italy. The species were the indigenous Austropotamobius pallipes (the white-clawed crayfish) and Potamon fluviatile (the river crab) and the alien Procambarus clarkii (the red swamp crayfish, in Italy aquacultured since 1977 and invasive since 1989). Water and sediments from the three environments in which they occur were analysed for trace metals to evaluate the degree of contamination of the habitats. Procambarus clarkii showed a significantly higher accumulation of most of the metals analysed, including the non-essential cadmium, nickel and lead. Interspecific differences were not related to exposure to higher concentrations of metals, but indigenous and alien macro-decapods possibly differ in their physiological requirements for essential trace metals and/or in their rates of essential and non-essential metal uptake, accumulation, detoxification and excretion.     

Grazing by a native and an exotic crayfish on aquatic macrophytes

1. We compared grazing by native noble crayfish ( Astacus astacus ) and the exotic signal crayfish ( Pacifastacus leniusculus ) on seedling or well-established macrophytes.
2. In a pool experiment, seedlings of emergent Scirpus lacustris and floating-leaved Potamogeton natans were heavily grazed by adult signal crayfish, whereas established plants of the same species sustained only minor damage.
3. In a preference experiment two submerged macrophytes ( Chara vulgaris and Elodea canadensis ), and both seedlings and established plants of S. lacustris and P. natans , were presented pairwise to signal and noble crayfish. There was no significant difference in preference by the two crayfish species. Chara vulgaris was preferred to all other plants presented, established plants of S. lacustris and P. natans were never preferred, and seedlings of P. natans were preferred to established P. natans .
4. An aquarium experiment was conducted in which the consumption of Chara by signal and noble crayfish was measured in relation to water temperature. Signal crayfish consumed significantly more Chara than noble crayfish, especially at higher temperatures.
5. Our results indicate that the signal crayfish is the more voracious grazer especially at higher temperatures. There may be negative effects on vegetation (emergents and floating-leaved, as well as submerged species) when the signal crayfish is introduced. Chara species are particularly susceptible, since they are preferred by crayfish and the genus includes a large number of rare species. Stocking of crayfish therefore could lead to the decline or removal of submerged species in order of crayfish feeding preferences and could prevent the spread or cause a decline of emergent and floating-leaved vegetation.     

不同抗应激反应能力克氏原螯虾肝胰腺的代谢组分析

为探讨不同抗应激反应能力克氏原螯虾体内代谢物的差异, 研究通过运输应激和温度应激处理后, 选取抗应激反应能力强(SSR)和抗应激反应能力弱(WSR)的克氏原螯虾, 取肝胰腺, 通过液相色谱-质谱/质谱(LC-MS/MS)进行代谢组学分析。质谱共检测到10292个离子, 从中筛选、鉴定出了464个显著差异的代谢物(差异倍数>1.20 或 <0.83,P<0.05, 且 VIP>1.0), 其中与WSR相比, 在SSR中下调的代谢物227个, 上调代谢物237个。KEGG分析显示, 这些差异代谢物主要富集在氨基酸代谢通路, 包括组氨酸代谢、牛磺酸和亚牛磺酸代谢、赖氨酸降解、缬氨酸和亮氨酸及异亮氨酸生物合成、谷胱甘肽代谢等, 同时也富集到抗坏血酸和醛酸盐代谢途径、碳水化合物代谢途径(戊糖和葡萄糖醛酸相互转化)和脂肪酸代谢途径(不饱和脂肪酸生物合成)等。这些结果表明, 克氏原螯虾在应对运输和温度应激时存在广泛的代谢应答, 其中一些与抗氧化应激和增强免疫力相关的代谢物, 如γ-L-谷氨酰-L-半胱氨酸、牛磺酸和油酸等可能在抗应激反应过程中发挥重要作用。研究不仅可为动物抗逆境机制的研究提供新的思路, 而且在克氏原螯虾优良品种的培育及寻找应对克氏原螯虾应激反应的策略等方面也具有重要价值。     

Asynchronous swimmeret beating during defense turns in the crayfish, Procambarus clarkii

Swimmeret beating was monitored in freely moving specimens of the crayfish Procambarus clarkii as they exhibited defense turn responses to tactile stimuli. Analysis of videotape records revealed alterations in swimmeret beating during turning responses compared to straight, forward walking. During turns, swimmerets beat with shorter periods and smaller amplitude power strokes than during straight walking. Coordination between swimmerets also changed. Swimmerets on the side toward which the animal turned tended to lag behind their contralateral partners, rather than beat in synchrony as in straight walking, and ipsilateral coordination was loosened relative to straight walking. Asynchronous swimmeret beating accompanied asymmetric motions of the uropods in a manner similar to that observed during statocyst-dependent equilibrium reactions in P. clarkii, but removal of the statoliths did not eliminate turn-associated responses of the swimmerets. The coordinated action of the swimmerets and uropods may contribute to the torque that rotates the animal in the yaw plane. Implications of the observed changes in swimmeret coordination for understanding the underlying neuronal control system are discussed.     

Pike and red swamp crayfish: a new case on predator-prey relationship between aliens in central Spain

Pike in the Ruidera Lakes (central Spain) chiefly ate the recently introduced crayfish Procambarus clarkii . It was the dominant prey in occurrence, number and biomass for every size class and season. Likewise, number (up to 27) and total weight of ingested crayfish were directly related with pike size. The principal prey fish were all introduced species (except the endangered Blennius fluviatilis ), most native species having disappeared since pike were introduced in 1953. (c) 1996 The Fisheries Society of the British Isles     

Bioenergetics and phenology of reproduction of the introduced red swamp crayfish, Procambarus clarkii, in Do·ana National Park, Spain, and implications for species management

1. The red swamp crayfish, Procambarus clarkii (Girard), is a North American species which was introduced into the Lower Guadalquivir, Spain, in 1974. Procambarus clarkii has become a key species since it has greatly altered the functioning and structure of the ecosystems it has invaded.
2. Since P. clarkii spawn spontaneously throughout the year, it is to be expected that their ecological impact on the ecosystem may vary depending of the timing of habitat flooding (e.g. temporary freshwater marsh, and temporary or permanent pond). The objective of the present study was to understand the reproductive phenology and bioenergetics of P. clarkii in Do·ana National Park, Spain, freshwater ecosystems, and consequently, propose some management plans for their populations.
3. Five stages of ovarian development were detected. Final ovarian maturation occurred a few days before spawning and spawning was synchronized among mature females of all populations. Egg numbers hatched in pleopods ranged from 450 to 550. Timing of juvenile release from pleopods varied greatly (10–35 days).
4. Gravid females support most of their metabolic needs by catabolizing carbohydrates (overall mean of O:N = 30.39), and to a lesser extent, lipids. The aerobic and excretory metabolisms of gravid females were lower than those of immature animals of similar weight.
5. Analysis suggested that several factors influence the spawning of crayfish at the monitored sites. The major factors are water regime and temperature. Red swamp crayfish appear to be a spontaneous spawning species which may adjust their reproductive cycle to the timing of flooding, enhancing the utilization of available resources.
6. Populations of P. clarkii in Do·ana National Park could be controlled by the management of the water regime and by devising a fishing scheme which would maintain an appropriate population structure.     

Evidence of female cryptic choice in crayfish

To test whether male body size affects female reproductive investment in the polygamous crayfish Procambarus clarkii, we described mating behaviour of virgin females paired with either small or large males, and analysed the number, size and weight of both eggs and juveniles sired by either types of male. Along with confirming the overt selection by females of larger mates, we found that the size and weight of both the eggs and the juveniles were higher when sired by larger fathers. This suggests that P. clarkii females exert a form of cryptic choice for large males, seemingly adjusting the quantity of egg deutoplasm in function of the mate body size. The question of why females spend time and energy to brood low-fitness offspring is finally raised.     

Managing invasive crayfish: use of X-ray sterilisation of males

1. In the last decade, many attempts have been made to contain the spread of the North American crayfish Procambarus clarkii , an invader of many European water bodies, but none has been successful.
2. This study investigates the effects that ionising irradiation has on the reproduction of P. clarkii males by analysing the behaviour of the treated individuals, their reproductive output, the vitality and survival of their offspring and the damages induced to their gonads.
3. We found that a dose of 20 Gy X-rays did not compromise either the survival or the mating ability of males, but reduced the size of their testes and significantly altered spermatogenesis. The number of aborted eggs was larger in clutches sired by treated males, with a consequent reduction (by 43%) in the number of offspring.
4. These results foster hopes in our efforts to control invasive crayfish, showing the potential effectiveness of ionising irradiation to reduce male fertility in the perspective of adopting the Sterile Male Release Technique to control P. clarkii populations.     

Peculiarities of Cardiac Activity in the Crayfish Astacus astacus in Different Functional States

Analyzed are changes of temporal and amplitude characteristics of the cycles in animals in different functional states, using a non-invasive method of recording of cardiac activity in the adult noble crayfish Astacus astacus L. under laboratory conditions. It has been shown that the state of excitation in the crayfish after action of an intense stimulus lasts for a long time (more than 1 h) and can have no manifestations in some observed behavioral responses. Based only on the cardiac response (the heart rate), the real physiological state of the animal could be evaluated. Characteristic of the excited animal is a short-term response to the stimulus. In a high excitement of the animal (a high heart rate), significant transformations of the heart activity in response to other current stimuli may be barely noticeable. For the quiet animal, the response to a stimulus is longer than for the excited animal at the same stimulus intensity; for a comparative search for significant readjustments of the animal state, based on the heart activity parameters, the cardiograms typical of the quiet animal should be used. In the quiet animals, the cardiogram parameters change to a greater extent than in the excited ones. The quiet animals have longer diastole duration (DD) than the systole duration (SD). The diastole amplitude (DA) is higher than the systole amplitude (SA), and DA changes more markedly than SA. The healthy animals are characterized by equal values of DD and SD, sometimes by longer SD than DD, but, on the background of a continuing action, their values and the values of DA and SA become equal. There were noted a negative correlation between the systolic and diastolic phases by temporal parameters for healthy animals and an increase of the correlation significance in transition of the animal into the excited state. Based on the cardiovascular system response, the chief features of a shift of the functional crayfish state to pathological direction seem to include a stable heart rate reduction, a decrease of the heart contraction amplitude, a more frequent predominance of SD over DD, an increase of the means and the standard deviations of the parameters, especially of the cycle amplitudes. Changes of the sign and significance of correlation relations between inotropic and chronotropic parameters of cardiac activity in sick animals appear to indicate a disturbed mechanism of central coordination of the cardiovascular system work with its autonomic mechanisms. The results are discussed on the basis of known mechanisms of the heart activity control in Decapoda. The records of cardiac activity, such as laser light cardiograms (LLC), in testaceous invertebrates may be useful and interesting in studies on mechanisms and processes of internal control and management of the cardiovascular system activity of the animals.     

Eleven polymorphic microsatellites isolated from red swamp crayfish, Procambarus clarkii

Eleven microsatellites were isolated from a genomic DNA library enriched for CA- and GA-repeats, and were characterized in 48 individuals of red swamp crayfish Procambarus clarkii. All 11 microsatellites were polymorphic with an average of allele number of 6.6 per locus. Genotypic distributions of these 11 markers were found to be in conformance with the expectations of Hardy-Weinberg equilibrium. All 11 markers were unlinked. These markers are being used to study the invasion routine, genetic diversity and population structure of the species P. clarkii.     

Heart and ventilatory measures in crayfish during environmental disturbances and social interactions

Most animals assess the environment in which they live and alter their behavior according to various stimuli. When the animal does not make significant behavioral changes, as measured by bodily movements, the animal may be characterized as unresponsive to a given stimulus. This study demonstrates that when behavioral movements of crayfish cannot be observed, physiological measures of heart rate (HR) and ventilatory rate (VR) show dramatic changes in response to defined sensory stimuli. In the majority of cases, upon anticipation of a social interaction with another crayfish both HR and VR will increase. During an agonistic encounter between two crayfish, the level of HR and VR correlate with the intensity of the interaction. Such rapid responses in cardiac and respiratory systems to environmental disturbances and anticipation of a social interaction suggest an autonomic-like regulation associated with fear, flight or fight. Since behavioral observations do not allow an internal status to be readily assessed, we suggest that HR and VR may serve as a useful bioindex in crustaceans to their internal drive or possibly an awareness level to environmental cues.