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.     

Effects of crayfish on leaf litter breakdown and shredder prey: are native and introduced species functionally redundant?

Recent increases in biological invasions frequency may have important consequences on native communities. However, functional redundancy between invasive and native species could reduce non-native species effects on native ecosystems. Despite this, even small differences in functional traits between these species may still have unpredictable effects on colonized ecosystems. Invasive crayfish, as ecosystem engineers, potentially have wide and complex effects on recipient ecosystems, even when replacing a native counterpart. We used laboratory microcosms to test whether native (Astacus astacus) and invasive crayfish species (Orconectes limosus, Pacifastacus leniusculus and Procambarus clarkii) are actually functionally redundant in their effects on prey/shredder density and leaf litter breakdown. Results show that crayfish strongly influenced macroinvertebrate numbers and leaf litter breakdown and indicate that differences in direct (prey and leaf litter consumption) and indirect (prey habitat use and leaf litter breakdown) effects between crayfish species do exist. While the replacement of A. astacus by O. limosus may have induced only minor changes in freshwater ecosystems, invasions by the larger and more aggressive P. clarkii and P. leniusculus will likely have strong effects on invaded ecosystem. Overall, there seems to be no functional redundancy between these four species and outcomes of crayfish invasion will likely be species specific.     

Plasticity in fecundity highlights the females’ importance in the spiny-cheek crayfish invasion mechanism

Invasion is one of the most consequential phenomena affecting the distribution of native species. Few in number of species, European crayfish are losing the competition with introduced North American crayfish. The spiny-cheek crayfish, Orconectes limosus, is an outstanding example, successfully competing against the native narrow-clawed crayfish, Astacus leptodactylus. For four years, we collected data regarding crayfish occurrences, their relative abundance, and the structure of populations in the ongoing colonisation process of O. limosus in the lower Danube. The mature females of both invasive and indigenous crayfish species were analysed with respect to biometry and production of oocytes in relation to the dynamics of invasion. The interspecific comparisons showed no significant differences regarding body size, with an average of approximately 102 mm total length and 31 g wet weight for both species. However, the fecundity of the indigenous species was found to be constant throughout the investigated area, whereas the number of eggs produced by the invasive females was significantly increased at the active front of the invasion. The maximum number of ovarian eggs found was 887 and 1156 in the indigenous species and the invasive species, respectively. We propose the scenario that the invasive species, which carries the deadly crayfish plague, creates an ecological advantage by reducing the populations of indigenous crayfish. Subsequently, the invasive females opportunistically use the available resources to enhance their fecundity, resulting in the acute growth of populations. However, the long-term competitiveness and colonisation success of O. limosus still remain in question.     

The role of the Y-organ and cephalic gland in ecdysteroid production and the control of molting in the crayfish,Orconectes limosus

Summary The production of ecdysteroids (monitored by RIA) by Y-organs and cephalic glands in vitro was measured and hemolymph ecdysteroid levels were determined in the crayfish,Orconectes limosus, both after eyestalk ablation and as a function of time during natural premolt. Y-organ synthesis of ecdysteroid increased in parallel with a rise in hemolymph ecdysteroid concentrations under both conditions, peaking in substage D₂ of premolt. Y-organ ecdysteroid output after eyestalk ablation was 3–4 times higher. Thus, removal of the inhibiting system of the eyestalk effectively removes not only the principal control but also any modulation of ecdysteroid secretion by the Y-organs. Ecdysteroid levels remained low in Y-organ-ectomized crayfish, although premolt was initiated in some animals. The cephalic gland does not appear to contribute to the regulation of molting inOrconectes limosus. The Y-organs, on the other hand, are a principal source of ecdysteroids which regulate the major synthetic activities of premolt.     

Species-specific pattern of crayfish distribution within a river network relates to habitat degradation: implications for conservation

The degradation of aquatic habitats has increasingly become one of the most important factors influencing the distribution of freshwater species worldwide. We analysed the occurrence of three crayfish species, Astacus astacus, Austropotamobius torrentium and Orconectes limosus, in relation to habitat degradation status (based on Directive 2000/60/ES), stream morphology, geographical characteristics and land cover. In total, we analysed 6,768 sites within the Czech Republic (Central Europe), of which 6,187 sites lacked crayfish; among the remainder, A. astacus was present in 507 sites, O. limosus occurred in 44 sites and A. torrentium was present in only 30 sites. The analysis revealed that A. astacus preferred streams of better water quality that were not surrounded by agricultural land or settlements. This species also preferentially occurred in smaller streams with stony bottom substrata that were located at higher altitudes. Austropotamobius torrentium occurrence was associated with the natural character of the water body and the presence of protected areas at higher altitudes. Conversely, the non-native crayfish species O. limosus was typically recorded at lower altitudes in downstream reaches surrounded by agricultural land and with deteriorated water quality. The degree of environmental (human) pressure besides differences in habitat characteristics at sites with ICS and NICS may be of general importance for conservation strategies aimed at central European native crayfish species, primarily because the habitat-driven co-occurrence pattern (and its possible changes in the future) may strongly influence interspecific relationships, such as direct competition and the spread of infectious diseases between crayfish species.     

Reproductive Plasticity in Freshwater Invader: From Long-Term Sperm Storage to Parthenogenesis

Orconectes limosus, a North American crayfish species, is one of the most important aquatic invaders in European inland waters. Despite more than 120 years occurrence in Europe and intense research, there are still gaps in knowledge of its life history and ecology. Investigation into O. limosus invasive success requires identifying the mechanisms that enabled them to establish dense and widespread populations from small initial numbers without observable limitation by an introduction bottleneck. In part, O. limosus success may lie in its ability to reproduce by facultative parthenogenesis. Moreover, there are possible other mating scenarios, because of two mating seasons (autumn and spring) in O. limosus. This work investigated the effect of four reproductive scenarios (autumn mating only, spring mating only, autumn and spring mating, and without mating) on the reproductive success of O. limosus. Females successfully reproduced in all tested mating regimes using parthenogenesis as well as log term sperm storage. This reproductive plasticity likely facilitates the overwhelming success of O. limosus spread and establishment in new localities. It can explain the spread of O. limosus from the initial introduction of 90 specimens to most of continental Europe and Great Britain. These conclusions imply a serious threat, not only for autochthonous European astacofauna, but for other aquatic organisms as well as entire ecosystems.     

Consumptive effects of fish reduce wetland crayfish recruitment and drive species turnover

Predators and dry-disturbances have pronounced effects on invertebrate communities and can deterministically affect compositional turnover between discrete aquatic habitats. We examined the effect of sunfish (Lepomis spp.) predators on two native crayfish, Procambarus alleni and P. fallax, that regionally coexist in an expansive connected wetland in order to test the hypotheses that sunfish predation limits crayfish recruitment (both species) and that it disproportionately affects P. alleni, the species inhabiting temporary wetlands. In replicate vegetated wetlands (18.6 m²), we quantified summertime crayfish recruitment and species composition across an experimental gradient of sunfish density. Separately, we quantified effects of sunfish on crayfish growth, conducted a complimentary predation assay in mesocosms, and compared behavior of the two crayfish. Sunfish reduced P. alleni summertime recruitment by >99% over the full sunfish gradient, and most of the effect was caused by low densities of sunfish (0.22–0.43 m⁻²). P. alleni dominated wetlands with few or no sunfish, but the composition shifted towards P. fallax dominance in wetlands with abundant sunfish. P. fallax survived better than P. alleni over 40 h in smaller mesocosms stocked with warmouth. Sunfish reduced P. fallax recruitment 62% in a second wetland experiment, but growth rates of caged crayfish (both species) were unaffected by sunfish presence, suggesting predatory effects were primarily consumptive. Consistent with life histories of relatively fish-sensitive invertebrates, P. alleni engaged in more risky behaviors in the laboratory. Our results indicate that sunfish predators limit recruitment of both species, but disproportionately remove the more active and competitively dominant P. alleni. Spatially and temporally variable dry-disturbances negatively co-varying with sunfish populations allow for regional coexistence of these two crayfish and may release populations of either species from control by predatory fishes.     

Crayfish assemblage shifts in a large drought-prone wetland: the roles of hydrology and competition

1. Faster growing, larger and/or more aggressive crayfish species are predicted to dominate permanent waterbodies. We tested this prediction using a 9 year dataset for two species of crayfish (Procambarus alleni and Procambarus fallax) co‐existing in a sub‐tropical flowing slough in southern Florida. Using a series of laboratory and mesocosm experiments we also compared life history traits and performance of the respective species to test mechanisms that could explain dominance shifts in the local crayfish assemblages. 2. Over the 9‐year period, P. alleni densities were the greatest in shallower, shorter‐hydroperiod areas bordering the slough, while P. fallax densities were higher in deeper, longer‐hydroperiod central areas. These areas were separated by 0.8–2 km of continuous wetland with no apparent barriers to movement between them. 3. Density of P. fallax was not strongly affected by any measures of hydrological variation, while P. alleni density increased with more severe drought conditions. Following the strongest droughts, P. alleni colonized areas in the centre of the slough where they had been absent or scarce in wetter years. 4. We conducted experiments to compare growth rates, drought tolerance, and competitive dominance of these species. P. alleni survived drought conditions better, had higher growth rates, and was the dominant competitor for space and food. While drought probably limits P. fallax in the drier slough habitats, neither drought sensitivity nor interspecific competition with P. fallax can explain decreases of P. alleni with wetter conditions. 5. Our results indicate that a competition‐colonization tradeoff cannot explain the crayfish compositional dynamics in this wetland because P. alleni is both the best competitor and the best at surviving in and colonizing areas with the strongest droughts. Future attention should focus on the potential for selective effects of predators that co‐vary with hydrology. 6. The traits (large size, fast growth, competitive dominance) exhibited by P. alleni, which is absent in long‐hydroperiod wetlands, are those exhibited by dominant crayfish in permanent lakes and streams containing fish. Although these traits make crayfish less vulnerable to fish in some lakes and streams, life‐history models of community structure across permanence gradients suggest the opposite traits should be favoured for co‐existence with fish.     

Prevalence of the Crayfish Plague Pathogen Aphanomyces astaci in Populations of the Signal Crayfish Pacifastacus leniusculus in France: Evaluating the Threat to Native Crayfish

Aphanomyces astaci, the crayfish plague pathogen, first appeared in Europe in the mid-19^th century and is still responsible for mass mortalities of native European crayfish. The spread of this parasite across the continent is especially facilitated by invasive North American crayfish species that serve as its reservoir. In France, multiple cases of native crayfish mortalities have been suggested to be connected with the presence of the signal crayfish Pacifastacus leniusculus, which is highly abundant in the country. It shares similar habitats as the native white-clawed crayfish Austropotamobius pallipes and, when infected, the signal crayfish might therefore easily transmit the pathogen to the native species. We investigated the prevalence of A. astaci in French signal crayfish populations to evaluate the danger they represent to local populations of native crayfish. Over 500 individuals of Pacifastacus leniusculus from 45 French populations were analysed, plus several additional individuals of other non-indigenous crayfish species Orconectes limosus, O. immunis and Procambarus clarkii. Altogether, 20% of analysed signal crayfish tested positive for Aphanomyces astaci, and the pathogen was detected in more than half of the studied populations. Local prevalence varied significantly, ranging from 0% up to 80%, but wide confidence intervals suggest that the number of populations infected by A. astaci may be even higher than our results show. Analysis of several individuals of other introduced species revealed infections among two of these, O. immunis and P. clarkii. Our results confirm that the widespread signal crayfish serves as a key reservoir of Aphanomyces astaci in France and therefore represents a serious danger to native crayfish species, especially the white-clawed crayfish. The prevalence in other non-indigenous crayfish should also be investigated as they likely contribute to pathogen transmission in the country.     

Enhanced ecological indication based on combined planktic and benthic functional approaches in large river phytoplankton ecology

Environmental DNA (eDNA) is a powerful method for assessing the presence and distribution of invasive aquatic species. We used this tool to detect and monitor several invasive crayfishes Procambarus clarkii, Orconectes limosus and Pacifastacus leniusculus present in, or likely to invade, the ponds of the Brenne Regional Natural Park. A previous study showed that the eDNA method was not very efficient in detecting P. clarkii. In the present study, we explored new improvements in the detection of invasive crayfish. We designed specific primers for each crayfish species, and set up an experimental mesocosm approach to confirm the specificity of the primers and the sampling protocol. We analysed samples taken from ponds in 2014 and 2015. We compared two qPCR protocols involving either SybrGreen or TaqMan assays. Using these same primers, we were able to detect crayfish eDNA with both assays during the mesocosm experiment. However, crayfish from field samples could only be detected by performing qPCR with a SybrGreen assay. We successfully monitored the presence of three invasive species of crayfish using eDNA. This method is a powerful tool for establishing the presence or absence of invasive species in various freshwater environments.     

A local gammarid uses kairomones to avoid predation by the invasive crustaceans Dikerogammarus villosus and Orconectes limosus

Since the 1980s the Ponto-Caspian gammarid Dikerogammarus villosus has spread throughout Europe while displacing native species and is predicted to invade further continents. After it was introduced into Europe in the 1890s the North American crayfish Orconectes limosus spread throughout Europe and served as a vector to displace native crayfish as well. In Lake Constance (Germany) the previously dominant gammarid Gammarus roeselii is subjected to both of these invasive crustaceans. In our experiments both species placed predation pressure on G. roeselii. Kairomone perception tests in a Y-maze revealed the capability of the gammarids G. roeselii and D. villosus to perceive and avoid the scent of the predator crayfish O. limosus. Both species also avoided the kairomone of the other gammarid, but did not avoid the scent of their own species. This taxa specific behavior suggests that taxa specific signals are used. This behavior can help the gammarids avoid shelters previously occupied by predators.     

Trophic niches of three sympatric invasive crayfish of EU concern

The spread of non-native species results in novel and often unexpected assemblages. Using stable isotopes, we disentangled the trophic relationships between three invasive crayfish species at two sites of a small thermal tributary of the Barát stream, Hungary. We studied Procambarus virginalis and Faxonius limosus living in sympatry in the upper section of this thermal tributary, and then an assemblage in a lower section also containing P. clarkii. The two species in the upper section largely shared trophic niches, although P. virginalis was more carnivorous than F. limosus, which fed more on detritus and aquatic plants. In the lower section, P. clarkii had a distinctive trophic niche, being more carnivorous than the other species and also preying on other crayfish and fish. The trophic niches of the other two species shifted slightly, being narrower and more overlapping in the presence of P. clarkii. It seems that the presence of P. clarkii affects the feeding habits and trophic niches of the other two crayfish. Our results also indicate that the species have somewhat distinctive feeding niches, which suggests that the ecosystem effects of these species are likely to be at least partially additive in the shared localities.

     

Invasive species of crayfish use a broader range of predation-risk cues than native species

The responses of invasive and native species of crayfish to conspecific and heterospecific alarm odors were recorded in the laboratory. Individuals of the North American invasive Procambarus clarkii responded just as strongly to odors from crushed Austropotomobius pallipes as they did to crushed conspecifics. The North American invasive Orconectes limosus also responded as strongly to P. clarkii odor as to conspecific odor. The native Italian species A. pallipes responded more strongly to conspecific alarm than to heterospecific alarm from P. clarkii. The pattern of invasive species of crayfish using a broader range of danger signals than displaced native species appears to be robust.     

Preventing overexploitation in a mutualism: partner regulation in the crayfish–branchiobdellid symbiosis

For a symbiosis to be a mutualism, benefits received must exceed costs incurred for both partners. Partners can prevent costly overexploitation through behaviors that moderate interactions with the other symbiont. In a symbiosis between crayfish and branchiobdellidan annelids, the worms can increase crayfish survival and growth by removing fouling material from the gills. However, overexploitation by the worms is possible and results in damage to host gills. We used behavioral observations to assess the degree to which two species of crayfish (Cambarus chasmodactylus and Orconectes cristavarius) use grooming to moderate their interaction with branchiobdellids. We found that grooming could effectively reduce worm numbers, and the proportion of total grooming directed at worms differed between crayfish species and as a function of worm number. O. cristavarius increased grooming in response to the addition of a single worm, while C. chasmodactylus only increased grooming in response to ten worms. These differences in the number of worms that trigger grooming behavior reflect differences between crayfish species in field settings. We also assessed whether antibacterial compounds in circulating crayfish hemolymph could limit bacterial gill fouling. O. cristavarius hemolymph inhibited some test bacteria more effectively than C. chasmodactylus did. Differences in the antibacterial properties of crayfish hemolymph may therefore help explain differences in both worm-directed grooming and worm loads in the field. We conclude that crayfish can use grooming to reduce worm numbers, which could lower the potential for gill damage, and that the level of grooming varies between crayfish species.     

The effect of inter- and intra-specific competition on survival and growth rate of native juvenile noble crayfish <Emphasis Type="Italic">Astacus astacus</Emphasis> and alien spiny-cheek crayfish <Emphasis Type="Italic">Orconectes limosus</Emphasis>

Juvenile noble crayfish, Astacus astacus and spiny-cheek crayfish, Orconectes limosus were reared from the 2nd and the 3rd developmental stage (i.e. since the stage of independence), respectively, until the end of the growing season. The first period revealed very low survival rate of O. limosus in the communal stock (7 ± 6.2%), probably caused by predation of A. astacus. Separately reared O. limosus proved significantly higher survival rate (49 ± 9.2%) than those in the communal stock with A. astacus. The latter species also showed the highest survival rate in this case (90 ± 7.1%). This was not found to differ from the survival rate of separately reared noble crayfish (86 ± 3.1%). In general, both the significant effect of bigger size of A. astacus than that of O. limosus and higher intra-specific cannibalism of O. limosus were reflected during the first phase of rearing. Size and weight of both species were equivalent 11 weeks after hatching (8 weeks of rearing). The higher cumulative survival rate was reached in the intraspecific culture of A. astacus (90%) while low cumulative survival of both species was found in communal stock (less than 50%) in the second phase of rearing. At the end of the growing season, 19-week-old O. limosus attained a higher mean size and weight but were more variable in the group (29.3 ± 5.17 mm and 616.4 ± 412.53 mg) than A. astacus (21.2 ± 1.82 mm and 229.9 ± 59.45 mg). Single and communal stock did not differ. Growth of O. limosus was much faster than that of A. astacus.     

Risk assessment of the crayfish pet trade based on data from the Czech Republic

The pet trade in freshwater crustaceans, including crayfish, has grown rapidly in recent decades and become an important pathway for introducing new non-indigenous species into Europe. This paper provides the first overview of non-indigenous crayfish species (NICS) traded as ornamental and their potential impact in the Czech Republic, which is the second leading importer into Europe. The paper presents a full list of traded crayfish species, their market availability, and trade names or misnomers used in the country. In total, 27 crayfish species from all three families are advertised and marketed, of which Astacus astacus is the only indigenous species. Only three NICS were recognized as very common on the market. The invasiveness and risk associated with ornamental crayfish trade were assessed using the Freshwater Invertebrate Invasiveness Scoring Kit. Five NICS were classified into the high-risk category, the highest score being for Procambarus fallax f. virginalis. The invasiveness of crayfish indigenous to North America is significantly greater than that of crayfish from the rest of the world, and therefore regulation in this regard is recommended.     

Cross-species amplification of microsatellite markers in the invasive spiny-cheek crayfish (Orconectes limosus): Assessment and application

The North American spiny-cheek crayfish,Orconectes limosus (Cambaridae), endangered in its native range, is a widespread invasive species in European waters and conservationally important carrier of crayfish plague. However, its population structure is poorly known, and no informative genetic markers for the species are available. We tested cross-species transfer of microsatellite loci to spiny-cheek crayfish from 5 other crayfish species. Variability of 10 successfully amplifying loci derived from 4 species was then tested in 60 individuals ofO. limosus originating from 3 natural populations: the river Danube at Bogyiszló in Hungary, a pond in Stary Klíčov, and the brook Černovicky, both in the Czech Republic. The allele number within the populations ranged from 4 to 10 alleles per locus, while heterozygosity levels varied from 0.650 to 0.900 forHo and from 0.660 to 0.890 forHe. No linkage disequilibrium and no null alleles were detected. The selected markers are useful for assessing population structure, intraspecific variation, and paternity studies in spiny-cheek crayfish.     

Positive selection of beautiful invaders: long-term persistence and bio-invasion risk of freshwater crayfish in the pet trade

After interest in keeping crayfish (Crustacea, Decapoda, Astacida) in home aquaria peaked in the mid-2000s, the aquarium trade has become a novel introduction pathway for non-native crayfish species in Germany. Here, we provide an update on the ornamental crayfish trade approximately one decade after the ‘crayfish hype’ to explore the long-term implications in terms of bio-invasion risk. Specifically, species’ availability in e-commerce and potential invasiveness were assessed and compared to previous studies. Morphological and ecological traits of the offered species were compiled and related to their long-term availability (covering 2005–2015). In July 2015, a total of 31 online shops offered 28 crayfish species, which represents a decline of 24 % in species diversity compared to the late 2000s. The estimated rate of import of new species has considerably flattened and approaches pre-hype values (<1 species year^?1). However, the risk associated with the offered species, as assessed by a risk screening tool, has not decreased compared to the late 2000s. Long-term availability in the trade was primarily determined by bright coloration, the ability to reproduce under warm aquarium conditions, and a preference for lentic habitats. Species featuring such traits are likely to persist in the aquarium trade and include four high-risk species, most notably invasive and crayfish plague-carrying red swamp crayfish (Procambarus clarkii) and Marmorkrebs (Procambarus fallax f. virginalis). Persistent propagule pressure from aquaria has substantially contributed to the establishment of both species in Germany, stressing the need for more effective pathway management.     

Spiny-cheek crayfish Orconectes limosus carry a novel genotype of the crayfish plague pathogen Aphanomyces astaci

The oomycete Aphanomyces astaci causes mass mortalities of European crayfish. Different species of North American crayfish, original hosts of this parasite, seem to carry different strains of A. astaci. So far, four distinct genotype groups have been recognised using Random Amplification of Polymorphic DNA (RAPD-PCR). We succeeded in isolating A. astaci from the spiny-cheek crayfish Orconectes limosus, a widespread invader in Europe, and confirmed that this species carries a novel A. astaci genotype. Improving knowledge on the diversity of this parasite may facilitate identification of genotypes in mass mortalities of European crayfish, thus tracing the sources of infection.     

Bimodal annual reproductive pattern in laboratory-reared marbled crayfish

The marbled crayfish, Procambarus fallax f. virginalis, is a new research model and potent invader of aquatic ecosystems that reproduces by obligatory parthenogenesis. My data show that in captivity, it can reproduce throughout the year. However, when kept at constant 20 °C and natural photoperiod there were two prominent spawning maxima, one before the spring equinox and another one before the fall equinox. If temperature fluctuated between ~15 °C in winter and ~25 °C in summer, the first maximum was shifted beyond the spring equinox and the second maximum was shifted closer towards the fall equinox. These results indicate a bimodal annual rhythm for the reproduction in marbled crayfish that can be modulated by temperature. Comparison with P. fallax suggests that this rhythmicity was inherited from its sexually reproducing mother species. Potential consequences of my laboratory findings for wild populations of marbled crayfish in temperate and tropical regions are discussed. Since marbled crayfish can be kept in captivity for up to four years under a broad range of conditions, they offer the possibility to investigate endogenous circannual rhythms and their entrainment by environmental zeitgebers in decapod crustaceans.