Niche breadth and trophic diversity: feeding behaviour of the red swamp crayfish (Procambarus clarkii) towards environmental availability of aquatic macroinvertebrates in a rice field (Portugal)

The red swamp crayfish, Procambarus clarkii (Girard 1852), an alien species in Portugal, may have dramatic effects on aquatic communities by depleting all food resources available, just after its introduction. The purpose of this study was to evaluate, through the concept of niche breadth, whether the diet of this species reflected the temporal changes of aquatic macroinvertebrate availability once it is acclimated. Petraitis’ index of niche breadth and Herrera’s trophic diversity index were used to evaluate the trophic behaviour of P. clarkii towards available resources over time and intraspecifically (size classes and sex) in a rice field in Portugal. Results from this study showed that the consumption of aquatic macroinvertebrates by P. clarkii reflected their seasonal availability. The high values of niche breadth and trophic diversity indicated resource use according to trophic availability, diversified diets and different individual exploitation of resources regardless of size or sex. These findings suggest that in habitats where P. clarkii is already acclimated, it adjusted its trophic behaviour to the seasonal availability of aquatic macroinvertebrates. The large niche breadth and high trophic diversity presented by P. clarkii enables it to successfully expand its range to new areas, when other environmental conditions are favourable, as has been observed in Portugal and worldwide.     

Predator–prey interactions of Procambarus clarkii with aquatic macroinvertebrates in single and multiple prey systems

Understanding the interspecific interactions of Procambarus clarkii with other aquatic macroinvertebrates will help to unveil the mechanisms and processes underlying biological invasiveness. The purpose of this study was to investigate predator–prey interactions of two ontogenic phases of P. clarkii with native and exotic species of aquatic macroinvertebrates at a single and multiple prey level. We performed laboratory experiments to determine the consumption and the behavioral responses of Chironomus riparius, Physa acuta and Corbicula fluminea to P. clarkii. The presence of P. clarkii significantly affected the abundance of C. riparius and P. acuta, but not of C. fluminea whether prey species were provided singly or simultaneously. The consumption of C. riparius by P. clarkii was higher than P. acuta for both crayfish sizes and situations (single/multiple prey systems) and C. fluminea was never consumed. Physa acuta was the only species that exhibited an anti-predator behavior to P. clarkii. Our results show that P. clarkii can have strong consumptive and trait effects on aquatic macroinvertebrate prey at a single and multiple prey level, resulting in differential impacts on different prey species. This study clarifies some aspects of the predator–prey interactions between P. clarkii and native as well as other exotic macroinvertebrate species that have invaded freshwater biocenosis worldwide.     

Impact of an introduced Crustacean on the trophic webs of Mediterranean wetlands

Based on a review and our own data, we present an overview of the ecological impacts on the trophic web of Mediterranean wetlands by an introduced Decapod Crustacean, the red swamp crayfish (Procambarus clarkii). P. clarkii lacks efficient dispersal mechanisms but is very well adapted to the ecological conditions of Mediterranean wetlands (fluctuating hydroperiods with regular intervals of drought). As an opportunistic, omnivorous species, which adapts its ecology and life history characteristics, such as timing and size at reproduction to changing environmental conditions, it became readily established in most of the Mediterranean wetland environments. High reproductive output, short development time and a flexible feeding strategy are responsible for its success as an invader. Like most crayfish, it occupies a keystone position in the trophic web of the invaded system and interacts strongly with various trophic levels. It efficiently grazes on macrophytes and is one of the main factors, besides the impact of flamingos, cattle and introduced fish, of the change of many water bodies from a macrophyte dominated, clear water equilibrium to a phytoplankton driven turbid water balance. Juveniles feed on protein rich animal food with the corresponding impact on the macroinvertebrate community in competition with other crayfish or fish species. At the same time, it serves as a prey for mammals, birds and fish. Due to its predatory and grazing activity, it efficiently canalises energy pathways reducing food web complexity and structure. Feeding also on detritus it opens, especially in marshlands, the detritic food chain to higher trophic levels which results in an increase of crayfish predators. As a vector of diseases, it has a severe impact on the preservation and reintroduction of native crayfish. P. clarkii accumulates heavy metals and other pollutants in its organs and body tissues and transmits them to higher trophic levels. Due to the long history of its presence, the complex interactions it established within the invaded ecosystems and the socio-economic benefits it provides to humans, prevention and control seem the most promising management measures to reduce the negative impact of this crayfish species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Home-field advantage: native signal crayfish (Pacifastacus leniusculus) out consume newly introduced crayfishes for invasive Chinese mystery snail (Bellamya chinensis)

The introduction of non-indigenous plants, animals and pathogens is one of today’s most pressing environmental challenges. Freshwater ecologists are challenged to predict the potential consequences of species invasions because many ecosystems increasingly support novel assemblages of native and non-native species that are likely to interact in complex ways. In this study we evaluated how native signal crayfish (Pacifastacus leniusculus) and non-native red swamp crayfish (Procambarus clarkii) and northern crayfish (Orconectes virilis) utilize a novel prey resource: the non-native Chinese mystery snail (Bellamya chinensis). All species are widespread in the United States, as well as globally, and recent surveys have discovered them co-occurring in lakes of Washington State. A series of mesocosm experiments revealed that crayfish are able to consume B. chinensis, despite the snail’s large size, thick outer shell and trapdoor defense behaviour. Crayfish exhibited size-selective predation whereby consumption levels decreased with increasing snail size; a common pattern among decapod predators. Comparison of prey profitability curves—defined as the yield of food (weight of snail tissue) per second of feeding time (the time taken to crack the shell and consume the contents)—suggests that small and very large snails may represent the most profitable prey choice. By contrast, previous studies have reported the opposite pattern for crayfish consumption on thin-shelled snails. For all snail size classes, we found that native P. leniusculus and invasive O. virilis consumed greater numbers of snails than invasive P. clarkii. Moreover, P. leniusculus consistently handled and consumed snails at a faster pace compared to both invasive crayfishes across the range of snail sizes examined in our study. These results suggest not only that B. chinensis is a suitable food source for crayfish, but also that native P. leniusculus may ultimately out-consume invasive crayfishes for this new prey resource.     

Crayfish invading Europe: the case study of Procambarus clarkii

The red swamp crayfish, Procambarus clarkii, native to northeastern Mexico and southcentral USA, is today the dominant macroinvertebrate in several European countries. While the first introduction of this species into Spain is well-documented, little is known about its pathways of invasion and the reason for its rapid spread in several European countries. Study of the biology of the species has revealed a number of properties that makes this crayfish a successful invader. Procambarus clarkii exhibits properties characteristic of an r-selected species, including early maturity at small body size, rapid growth rates, large numbers of offspring at a given parental size, and relatively short life spans. It is also plastic in its life cycle, able to disperse widely in the habitat and to tolerate environmental extremes. It displays generalist and opportunistic feeding habits, consuming macrophytes and preying on amphibians. Procambarus clarkii can also replace indigenous crayfish by a combination of mechanisms, including competitive exclusion and transmission of the fungus-like Aphanomyces astaci, responsible for the crayfish plague. Finally, this species shows a wide behavioral flexibility when coping with new types of predators. The results of these studies, combined with the increasing information available in the scientific literature on this and other crayfish species, will help us understand invasions in this taxon and make predictions about the identity of future crayfish invaders.     

Early assessment of the impact of alien species: differential consequences of an invasive crayfish on adult and larval amphibians

Aim Early assessment of the impact of invasive alien species is crucial to set up timely management, but often the impact is evident when it is too late for action. We evaluated relationships between the alien crayfish, Procambarus clarkii, the distribution of native amphibians, and the abundance of their larvae. We assessed whether considering measures of reproductive success provide a more prompt measure of impact than considering just species distribution. Location One hundred and twenty‐five wetlands in Northern Italy, in an area recently invaded by P. clarkii. Methods We surveyed wetlands to assess the presence of breeding activity of amphibians and the distribution of P. clarkii. We measured the abundance of amphibian larvae before metamorphosis through pipe sampling. We built models analysing the relationships between amphibian and crayfish distribution, while taking into account spatial autocorrelation and environmental features. Analyses were performed at both the species level (generalized linear models and spatial eigenvector mapping) and community level (constrained redundancy analysis). Results In terms of breeding site distribution, only two amphibians (Lissotriton vulgaris and Hyla intermedia) were negatively associated with P. clarkii, while the relationships between other the species and P. clarkii were positive or not significant. However, larval abundance for all amphibian species was negatively associated with the alien crayfish. Analyses performed at community and single species levels yielded consistent results. Main conclusions Procambarus clarkii impacts amphibians through different processes. Newts probably avoid invaded wetlands for breeding. Other species attempt breeding in wetlands with crayfish, but suffer very low success. Considering distribution data alone would not provide a correct picture of the impact of this alien species; measures of reproductive success may allow a more accurate assessment of the impact.     

Invasive crayfish in Europe: the impact of Procambarus clarkii on the littoral community of a Mediterranean lake

1. Despite the growing number of non‐indigenous crayfish species introduced worldwide, little scientific attention has been given to their effects on the recipient communities. Here, we quantified the impact of adult red swamp crayfish (Procambarus clarkii) on key components of the littoral community of a mesotrophic lake in central Italy. We used two densities of crayfish plus a no‐crayfish control, and two complementary methods –in situ enclosures and a laboratory study. 2. Both experiments showed that P. clarkii strongly affects the community even at a low density (4 m⁻²), although a high crayfish density (8 m⁻²) showed a more pronounced impact in a shorter time. 3. In enclosures, P. clarkii quickly consumed the pond snail Haitia acuta, notwithstanding its anti‐predator behaviour, but not the mosquitofish Gambusia affinis. The biomass of the hydrophytes Nymphoides peltata and Potamogeton spp. was strongly reduced by crayfish grazing, coupled with their non‐consumptive plant clipping and uprooting, which were particularly intense in the case of Potamogeton crispus. In contrast, Utricularia australis was avoided by P. clarkii, even when other resources were exhausted. 4. The introduction of P. clarkii could reduce biodiversity and increase biotic homogenisation in a short time.     

Colonization of freshwater habitats by an introduced crayfish, <Emphasis Type="Italic">Procambarus clarkii</Emphasis>, in Southwest Iberian Peninsula

The introduction of some crustacean species has produced alterations of freshwater environments and declines of native species worldwide. The red swamp crayfish, Procambarus clarkii Girard, was introduced in the Southwest Iberian Peninsula in the 1970’s, producing severe impacts on rice agriculture and on native biota such as macrophytes, gastropods, native crayfish and amphibians. We studied the distribution of P. clarkii in two areas of SW Iberian Peninsula: the Sado River basin (SW Portugal), an area colonized by this species around 1990, and the Parque Natural del Entorno de Doñana (SW Spain), colonized soon after its introduction in the Iberian Peninsula, in the 1970’s. Our main goal was to determine which factors limit crayfish distribution, which could help to identify the most effective management practices to contain its spread. Procambarus clarkii was found in most types of water bodies, including small and shallow ones. Distance to a crayfish source was the single predictor variable explaining crayfish occurrence in most types of habitats and in both areas. The only exception was for the Sado permanent stream points, where crayfish presence was negatively affected by an interaction between elevation and flow velocity. Other habitat characteristics have apparently little or no importance for its successful colonization. Moreover, this study indicated that overland dispersal is apparently a frequent phenomenon in this species. Our findings can be used to determine which habitats are most likely to be colonized by the crayfish and to develop practical measures which may limit its spread and minimize its impacts.     

Invasion of the shelter snatchers: behavioural plasticity in invasive red swamp crayfish,Procambarus clarkii

1. In many freshwater systems, competition for shelter plays an important role in determining the persistence of both native and alien species. The red swamp crayfish, Procambarus clarkii, is currently invading the native habitat of the signal crayfish, Pacifastacus leniusculus, in southern Oregon, and interspecific competition for shelter may be driving the species replacement in this region. 2. We designed a 2 × 3 factorial mesocosm experiment, with shelter density and species combination as factors, to investigate shelter occupancy and resource competition. Contrary to our predictions, the two crayfish species are equal competitors for shelter. Further, the invasive P. clarkii modified its shelter occupancy behaviour in the presence of the native P. leniusculus and has broader microhabitat preferences. 3. Specifically, we found that P. clarkii alters shelter occupancy and space use patterns when the two species occurred together, such that shelter use was identical between P. clarkii and P. leniusculus in mixed‐species treatments. In such treatments, both species increased their use of shelters when shelter density increased. When P. clarkii was alone, however, individuals did not alter shelter use as a function of shelter density, whereas P. leniusculus exhibited similar density‐dependent behaviour in both mixed‐ and single‐species treatments. 4. In a complementary field survey, we employed an ‘epicentre‐based’ design to sample two field sites. We observed patterns of microhabitat use and breadth for each species similar to those in our mesocosm experiment: the invasive P. clarkii was more abundant across different habitats and used a broader range of microhabitats than the native P. leniusculus. As such, we found that P. clarkii was more abundant across both field sites than the P. leniusculus, occupying microhabitats within and beyond the preferred range of P. leniusculus. Both field sites were affected by urban development and agriculture. 5. The use of microhabitats by both species was similar in the laboratory and the field. This study confirms that P. clarkii individuals can, and do, successfully occupy microhabitats preferred by P. leniusculus in the Willamette Valley. The results from our study may be relevant to other freshwater systems inhabited by P. clarkii and contribute to the understanding of ‘niche opportunity’, a concept which defines the environmental conditions that promote biological invasions.     

Can the parthenogenetic marbled crayfish Marmorkrebs compete with other crayfish species in fights?

The parthenogenetic marbled crayfish, Marmorkrebs, has no known wild population, but has been introduced into natural ecosystems in two continents. Interactions with native crayfish, particularly through fighting, could affect the ecological impact of Marmorkrebs introductions. Marmorkrebs have been characterized anecdotally as having low levels of aggression, which could mitigate their potential to compete with native species. We isolated Marmorkrebs and Louisiana red swamp crayfish (Procambarus clarkii), then conducted size-matched intra and interspecific pairings. Marmorkrebs were as likely to win a fight as P. clarkii, although contests between P. clarkii and Marmorkrebs began significantly faster than contests between two Marmorkrebs. These results suggest that Marmorkrebs have the potential to compete with other species on the same level as P. clarkii, which is itself a highly successful introduced species around the world.     

Is the exotic red swamp crayfish (Procambarus clarkii) a current threat for the Mediterranean tree frog (Hyla meridionalis) in the Camargue (Southern France)?

The non-indigenous red swamp crayfish (Procambarus clarkii) has been shown to be a threat for amphibian conservation. Many amphibian species breed in temporary ponds to diminish predation risk as such ecosystems are free of large predators. However P. clarkii, occurring as an invasive species in the Camargue delta, can readily disperse on the ground and thus colonize isolated ponds. We studied the current impact of the exotic crayfish on the reproductive success of the Mediterranean tree frog (Hyla meridionalis). In a mesocosm experiment, we tested the effect of two crayfish densities (1 and 3 crayfish/m²) on tadpole abundance. We also tested in a field experiment, within a temporary pond, the crayfish’s predation on the tree frog’s eggs. Finally, we developed site occupancy models using data from 20 ponds to assess the effect of crayfish abundance on tadpole abundance. Neither the experiments, nor the site occupancy models showed a negative impact of the current crayfish abundance on the tree frog populations breeding in ponds. We found that recorded crayfish densities were lower than in other areas where crayfish has impacted amphibian populations, but we hypothesize that current crayfish abundance in the area may increase in the future, thus impacting tree frog populations.     

Biological control of invasive populations of crayfish: the European eel (<Emphasis Type="Italic">Anguilla anguilla</Emphasis>) as a predator of <Emphasis Type="Italic">Procambarus clarkii</Emphasis>

The red swamp crayfish, Procambarus clarkii, is a paradigmatic invader of freshwater systems. Several attempts have been made to mitigate its multiple impacts but none was successful. Among the different methods proposed, the use of the European eel (Anguilla anguilla) as an indigenous predator is promising but the available information about its predatory ability on crayfish is to date scanty. To fill this gap in knowledge, we ran three experiments in wetlands and irrigation ditches in Italy. The first experiment, in the laboratory, was aimed at quantifying the extent of predation by eels on crayfish, the second, in enclosures, the size classes of crayfish mainly preyed and the possible effect of the eels on P. clarkii behaviour, and the third, in the field, its ability to effectively reduce crayfish populations. Results showed that eels prey on small-sized or soft crayfish, attacking them from the back; an indirect effect was to reduce crayfish trophic activity, which in turn might increases crayfish mortality due to starvation and decreases impact on the community. However, as shown in the field, the use of eels should be appropriately calibrated to the context of application. Taken together, our results show that eels might be used as a complement to the traditional trapping method. However, additional studies are necessary to understand the adequate number of eels to be introduced and to develop appropriate methods for quantifying such effects.     

Selective foraging by the crayfish Orconectes virilis and its impact on macroinvertebrates

SUMMARY. 1. The impact of crayfish predation on the abundance of macroinvertebrates was examined under semi-natural conditions. Female (Experiment 1) or male (Experiment 2) crayfish (Orconectes virilis) were held for 5 weeks in twelve small pools (4.67 m² surface area) at biomasses of 0. 5, 10 or 18 g m^?2 (live weight). The pools were stocked with known densities of macroinvertebrates. 2. Crayfish significantly affected the abundance of macroinvertebrates in the pools. Differences in the effects of crayfish on macroinvertebrates were related to crayfish sex, the presence of age-0 crayfish, and the species of macroinvertebrate. 3. The abundance of snails (Stagnicola elodes and Physa gyrina) was greatly reduced, in comparison with controls, by biomass of female crayfish ≥10 g m^?2 and by biomasses of male crayfish ≥5 g m^?2. The total density of non-molluscan invertebrates was inversely correlated with the biomass of female crayfish but the total biomass of non-molluscan invertebrates did not differ between treatments. This is consistent with our observation that small invertebrates (<2 mg wet weight) were less numerous, and large amphipods (32–64 mg) were more numerous, in pools stocked with female crayfish. In contrast, male crayfish had little apparent effect on the abundance of non-molluscan invertebrates. 4. Age-0 crayfish hatched at the end of Experiment 1 and were present in each pool at the start of Experiment 2. Surprisingly, male crayfish preyed little on age-0 crayfish. At the end of Experiment 2, the densities of age-0 crayfish varied between six and 116 individuals m^?2 and there was a strong inverse correlation between the mean biomass and density of age-0 crayfish recovered from the pools. This suggests age-0 crayfish were food limited in the pools and may explain the dominance of oligochaetes (which largely escape predation by burrowing) in the invertebrate community at the end of Experiment 2. 5. These results indicate that even relatively low densities of crayfish could greatly affect the abundance of macroinvertebrates in lakes. The introduction of crayfish into lakes (most lakes in Alberta currently have no crayfish) could substantially affect abundance and species composition of the macroinvertebrate community and, ultimately, the fish populations.     

Modeling mosquitofish (Gambusia holbrooki) responses to Genapol OXD-080, a non-ionic surfactant, in rice fields

An ecotechnological approach to control crayfish (Procambarus clarkii) infestation in rice fields of the Lower Mondego River Valley (Central Portugal) has recently been investigated. The application of the biodegradable non-ionic surfactant Genapol OXD-080, a fatty alcohol polyglycol ether, in rice paddies at a given concentration (50 mg/l) has been considered as a non-harmful chemical method to mitigate damage caused by crayfish digging activities to rice crops. Therefore, an important requirement regarding the ecological viability of this approach is that populations of non-target species are not significantly affected. A simple mosquitofish (Gambusia holbrooki) population model, in which the relationships with its main food prey were considered, was developed to assess the potential risk to a non-target key species of contaminating the irrigation channels following surfactant application. The model is based on data concerning mosquitofish life cycle and population dynamics, as well as mosquitofish diet and interactions with its main prey species. Quantitative information regarding the acute and sublethal effects of Genapol OXD-080 on mosquitofish and other non-target organisms was obtained from laboratory experiments. Three concentrations of Genapol OXD-080 (0.75, 1 and 2.5 mg/l) were used to simulate a small amount of contamination in irrigation channels. If contamination occurred, the mosquitofish population would tend to decline dramatically, even when submitted to a very small concentration of Genapol OXD-080 (e.g. 0.75 mg/l, 66.7 times lower than the concentration planned to be used in rice paddies). Thus, Genapol OXD-080 could potentially cause vast damage to local mosquitofish populations, and therefore should not be used without taking all precautions to avoid contaminating important biological reservoirs, such as the rice field irrigation channels.     

Loss of diversity and degradation of wetlands as a result of introducing exotic crayfish

The introduction of the alocthonous Louisiana red swamp crayfish (Procambarus clarkii) in Chozas (a small shallow lake situated in León (North-West Spain)) in 1996 switched the clear water conditions that harboured an abundant and a quite high richness of plants, invertebrates, amphibians and birds to a turbid one followed by strong losses in abundance and richness in the aforementioned groups. Crayfish exclusion experiments done in Chozas previous to this work confirmed the role of crayfish herbivorism on macrophyte destruction that had a trophic cascade effect on the wetland ecosystem. Direct and indirect effects of crayfish introduction on Chozas lake communities have been evaluated and compared with previous conditions before 1996 or with other related lakes in which crayfish were no present. Crayfish had a main role in submerged plant destruction and a potential effect on amphibia and macroinvertebrate population decrease. Plant destruction (99 % plant coverage reduction) was directly related to invertebrates (71 % losses in macroinvertebrate genera), amphibia (83 % reductions in species), and waterfowls (52 % reduction). Plant-eating birds were negatively affected (75 % losses in ducks species); nevertheless, fish and crayfish eating birds increased their presence since the introduction. Introduction of crayfish in shallow plant-dominated lakes in Spain is a main risk for richness maintenance in these endangered ecosystems. This revised version was published online in July 2006 with corrections to the Cover Date.     

Snail responses to cues produced by an invasive decapod predator

Abstract. Cues released by predators and injured prey often induce shifts in prey behavior that allow prey to evade predators, but also affect prey resource use. I investigated the effects of chemical and mechanical signals produced by injured snails (Physella gyrina) and predatory crayfish (Procambarus clarkii) on microdistributions of P. gyrina. In an initial experiment, I observed snail responses to the presence of a caged crayfish predator, to injured conspecifics, or to both. There were significant effects of time and the treatment × time interaction on the proportion of snails moving above the water line, with greater proportions of snails above the water line at night than during the day and with weak snail crawl‐out behavior being elicited by caged crayfish at night, but not during the day. In a second experiment, I examined snail microdistributions when exposed to crayfish confined to a small cage within each aquarium, crayfish confined to half of each aquarium, and crayfish ranging freely throughout each aquarium. Snails responded most strongly to free‐ranging crayfish by moving above the water line, but also demonstrated significant, but reduced, crawl‐out responses to crayfish confined to half of each aquarium; however, snails did not respond behaviorally to crayfish confined to small cages. In both experiments, there were marginally significant effects of unfed caged crayfish on the proportions of snail populations hiding under benthic shelters, with this response being the strongest at the start of the experiments but weak overall (with only 4–5% of P. gyrina responding in each experiment). These results indicate that cues (e.g., chemical or mechanical) produced by predators altered prey microdistributions, but that the exact prey responses (e.g., moving above the water line or into horizontal or benthic refugia) depended on the intensity and nature of cues.     

Evaluation of the suitability of the parthenogenetic marbled crayfish for aquaculture: potential benefits versus conservation concerns

The parthenogenetic marbled crayfish, Procambarus virginalis, is currently being discussed as a promising new candidate for aquaculture that could supply people in developing countries with high-quality protein and income. The main advantage of marbled crayfish is parthenogenetic reproduction. Comparison of growth between marbled crayfish and red swamp crayfish, Procambarus clarkii, the leading species in crayfish aquaculture revealed inferior body size and considerably slower growth in marbled crayfish. Only a very small proportion of the production would meet the size requirement of the international market and could serve as a cash crop. Aquaculture for local markets in extensive outdoor systems is probably economically feasible in developing countries, but the highly invasive and competitive marbled crayfish could easily escape from such sites, invade natural ecosystems and impair the autochthonous fauna and flora. Culture in closed indoor systems would be a safe alternative but this cost-intensive approach is economically not profitable. Because of small body size and slow growth, conservation concerns, and economic reasons, the marbled crayfish is considered unsuitable for aquaculture. It should not be spread around the globe for aquaculture and sustainable fisheries as was earlier done with the congeneric Procambarus clarkii, resulting in devastating ecological effects in numerous countries.

     

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

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Native European eels as a potential biological control for invasive crayfish

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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.