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All issues No 426 (2025) Knowl. Manag. Aquat. Ecosyst., 426 (2025) 25 Full HTML
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Knowl. Manag. Aquat. Ecosyst.
Number 426, 2025
Riparian ecology and management
Article Number 25
Number of page(s) 5
DOI https://doi.org/10.1051/kmae/2025020
Published online 07 October 2025

© J. Gao et al., Published by EDP Sciences 2025

Licence Creative CommonsThis is an Open Access article distributed under the terms of the Creative Commons Attribution License CC-BY-ND (https://creativecommons.org/licenses/by-nd/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. If you remix, transform, or build upon the material, you may not distribute the modified material.

Procambarus clarkii, originally native to northeastern Mexico and the south-central United States, is among the most globally invasive species (Oficialdegui et al., 2020). This species demonstrates a remarkable capacity for adaptation to low temperatures, facilitating its overwintering in temperate regions (Veselý et al., 2015). Its resilience in adverse conditions, including cold and drought, is largely attributed to its behavioral plasticity, particularly its burrowing behavior. Under extreme environmental stressors (e.g., drought, freezing temperatures), P. clarkii constructs burrows or migrates to new habitats, enabling the completion of critical life-history stages such as mating, egg incubation, and juvenile development (Gong et al., 2008; Souty-Grosset et al., 2014; Kouba et al., 2016). P. clarkii exhibits a broad thermal tolerance range of 5 °C to 38 °C (Cai et al., 2010). Its physiological adaptations, such as reduced heart rate and energy conservation, further enhance its survival under sub-freezing conditions (Kuklina et al., 2022).

Following its invasion, P. clarkii has become both abundant and frequently dominant across a wide range of aquatic habitats, including ditches, ponds, and streams. It is especially predominant in benthic species in river-connected lakes and floodplain wetlands, which are subject to seasonal fluctuations in water levels (Geiger et al., 2005; Foster and Harper, 2006). Its ecological success is largely attributed to a combination of factors: omnivorous and flexible feeding habits, broad ecological tolerance, high reproductive output, and, notably, its mobility and burrowing behavior, which allow it to adapt efficiently to varying hydrological conditions (Alcorlo et al., 2004; Gherardi, 2006; Alcorlo et al., 2008; Souty-Grosset et al., 2014). During seasonal low-water periods, P. clarkii burrows into exposed banks, marshes, or wetland substrates to avoid desiccation (Dong et al., 2008). These burrows offer essential shelter and help maintain humidity, providing a buffer against extreme environmental conditions (Herrmann and Martens, 2024). Anthropogenic activities, such as water extraction and dam regulation, along with climate change, have disrupted natural hydrological patterns, leading to altered timing and intensity of water-level fluctuations and drought events in floodplain ecosystems (Kingsford, 2000; Zhang et al., 2018). These changes may compel P. clarkii to modify its burrowing behavior and shelter-seeking strategies in response to new hydrological stressors. Despite its acknowledged adaptability, significant knowledge gaps persist concerning how P. clarkii adjusts to seasonal habitat changes to ensure successful overwintering. In particular, during periods of low temperatures combined with water-level declines, the availability of dynamic shelter (i.e., shelters whose accessibility changes with water-level fluctuations) may limit burrowing efficiency (Ilhéu et al., 2003; Barbaresi et al., 2004; Souty-Grosset et al., 2014). The degree to which these interacting stressors affect survival outcomes, including mortality risks due to unsuccessful burrowing, remains inadequately understood.

The present study seeks to examine the changes in the burrowing behavior of P. clarkii under varying temperature conditions (Autumn and Winter). It is well established that low winter temperatures reduce the overall mobility of P. clarkii (Gherardi et al., 2002a). We hypothesise that, compared to autumn conditions, the combined influence of water-level fluctuations and limited shelter availability under low winter temperatures will result in a more pronounced decline in burrowing capacity.

The study was conducted using 12 tapered plastic tanks (upper diameter: 85 cm, bottom diameter: 70 cm, height: 85 cm), each containing a 50 cm deep sediment layer. The tanks were positioned outdoors, and the sediment used in the experiment was collected from Lake Jinyin (30°63′−30°67′N, 114°17−114°23′E) in Wuhan City, China. It had a total nitrogen (TN) content of 0.79 mg g⁻1 and a total phosphorus (TP) content of 0.61 mg g⁻1. Before being introduced into the tanks, the sediment was air-dried, homogenized to ensure uniformity, and then added. According to the USDA soil texture classification (Giovani et al., 2019), the sediment comprised 27.56% clay, 72.44% silt, and 0% sand, which measured using a Malvern Mastersizer 2000 (0.02–2000 μm range) (Zha et al., 2022).

Four experimental microhabitat treatments were established in triplicate: (1) Constant Water Level without Shelter (CWL-NS): No shelters were provided, and a constant water depth of 25 cm above the sediment was maintained throughout the experiment; (2) Rapid Water Level Reduction without Shelter (RWLR-NS): No shelters were provided. Starting from an initial water depth of 25 cm, the water level was gradually reduced to 2 cm within 60 min following the introduction of the specimens; (3) Constant Water Level with Shelter (CWL-S): Shelters were provided, and a constant water depth of 25 cm above the sediment was maintained throughout the experiment; (4) Rapid Water Level Reduction with Shelter (RWLR-S): Shelters were provided. Beginning with an initial water depth of 25 cm, the water level was gradually reduced to 2 cm within 60 min following the introduction of the specimens. The rapid water level reduction (RWLR) treatment was designed to simulate conditions such as abrupt drought events or intentional dewatering of floodplain wetlands during the dry season.

Shelters were constructed using gray PVC 90° elbow joints (75 mm in diameter) to simulate natural refugia. In each tank with shelters, four shelters were evenly distributed across the sediment surface. Each tank (n = 12) was treated as an independent experimental unit, with two P. clarkii individuals (one male and one female) introduced per tank. The crayfish density used in this study was approximately 0.25 m2 per individual, which is consistent with the range adopted in previous studies on burrowing behavior, corresponding to approximately 0.24–0.38 m2 per individual (Haubrock et al., 2019a; Peeters et al., 2024). To differentiate between sexes, males were marked on the cephalothorax using non-toxic, acrylic-based, odourless white paint. The P. clarkii specimens were collected from natural ponds near Wuhan, China, and only individuals of uniform size were selected for the experiment. The mean body length and weight were 9.61 ± 0.64 cm and 29.18 ± 7.06 g for males and 9.85 ± 0.60 cm and 27.10 ± 5.47 g for females. Fifteen days before the experiment, the crayfish were acclimated in laboratory tanks that replicated the experimental conditions, with males and females housed separately to minimize pre-trial interactions. Crayfish were not fed during the experimental period to prevent potential interference of feeding activity with their behavioral responses.

The behavior of P. clarkii was monitored using high-definition infrared cameras equipped with night vision and automated periodic scanning (Video. S1). Behaviors were recorded in real-time and classified into five types based on the literature (Davis and Huber, 2007; Haubrock et al., 2019a): (1) Movement: Crawling with abdominal appendages; (2) Fighting: Engaging in combat behavior between two crayfish using claws; (3) Resting: Minimal activity, with only antennae and mouth movements; (4) Burrowing: Moving sediment with claws or tail; (5) Sideways Breathing: Lying on the side to breathe at the water surface. No activity was recorded when individuals remained inside burrows.

High-definition cameras were strategically positioned at fixed locations to monitor all 12 tanks. Each tank was filmed in sequence for 20-sec intervals, following a continuous cycle, with the predominant behavior during each sampling period being documented. The experiments were carried out under two distinct temperature regimes: mild-autumn conditions (18–23 °C, November 2023) and winter conditions (5–13 °C, January 2024). Each experiment lasted 72 h. The only difference between the two experiments was the variation in temperature.

The relative proportion of each behavior was calculated by dividing its duration by the total observation time and expressing the result as a percentage (Video. S1). To quantify overall activity levels under each treatment, activity was defined as the combined percentage of movement, fighting, and burrowing behaviors, providing a measure of P. clarkii's behavioral response to specific treatment conditions (Lozán, 2000). These metrics were calculated as follows equations (1) and (2):

Percentage of behaviour (%)=Duration of behaviourTotal duration of behaviour observations×100%(1)

Percentage of activity(%)=Duration of Movement+Duration of Fighting+Duration of BurrowingTotal duration of behaviour observations×100%(2)

The normality of behavioral data was assessed using the Shapiro–Wilk test, and the homogeneity of variance was evaluated with Levene's test. As the data did not meet the assumptions of normality and homogeneity of variance, the aligned rank transform (ART) method was used for nonparametric factorial ANOVA. This analysis included water level (i.e., RWLR and CWL), shelter (i.e., with or without artificial shelters), and season (i.e., autumn and winter) as fixed factors. When significant main effects or interactions were observed, pairwise comparisons were conducted using estimated marginal means. All statistical analyses were performed in R (version 4.4.2) using the ‘ARTool’ and ‘emmeans’ packages, with statistical significance set at p < 0.05 (Wobbrock et al., 2011).

The experimental results indicated that season had no significant effect on the burrowing behavior of P. clarkii (F = 1.51, p = 0.237), suggesting no difference in burrowing frequency between January and November. In contrast, activity levels were significantly higher in January than in November (F = 9.92, p < 0.01; post hoc: January > November, p < 0.01) (Tab. S1). In the RWLR-NS group, the number of burrows remained constant at 5 in both months. In the RWLR-S group, 4 burrows were recorded in November and 3 in January. For the CWL-S group, one burrow was observed in January, with none in November. Similarly, in the CWL-NS group, no burrows were observed in November, and only one was recorded in January (Tab. 1). As a poikilotherm species, P. clarkii generally exhibited reduced mobility in colder temperatures, which may limit their ability to seek or find suitable shelter (Haubrock et al., 2019b). However, P. clarkii exhibited a significantly higher survival rate in water temperatures (below 4°C) (Chucholl, 2011; Haubrock et al., 2019b). Despite this, P. clarkii demonstrates remarkable cold tolerance. Studies have shown that while acute low temperatures, such as a sudden drop from 21 °C to 5 °C, can significantly reduce heart rate, they do not lead to mortality (Chung et al., 2012). Moreover, P. clarkii, initially kept at 4 °C, began feeding after the water temperature was raised to 5°C, while individuals maintained at a starting temperature of 6 °C immediately consumed the offered prey (Haubrock et al., 2019b). Current research suggests that low temperatures do not limit the burrowing behavior of P. clarkii. The species' overwintering strategy integrates physiological resilience with behavioral adaptations. By constructing burrows, P. clarkii creates microhabitats that protect against freezing mortality in desiccating winter conditions.

This dual capacity for hypometabolic survival and sediment engineering allows for population persistence in hydrologically dynamic systems. Given the burrowing behavior of P. clarkii, which facilitates overwintering across all colonized habitats, its potential threat to river-connected lakes and floodplain wetland ecosystems may become more pronounced, particularly when the soil particle size composition is suitable for burrowing (Barnes, 2024). Experimental results revealed that burrowing activity was significantly reduced under the CWL treatment compared to the RWLR treatment (F = 21.04, p < 0.001; post hoc: CWL < RWLR, p < 0.001). Furthermore, burrowing behavior was significantly higher in environments without shelter compared to those with artificial shelter (F = 12.34, p < 0.01; post hoc: No shelter > Shelter, p < 0.01). A significant interaction between water level and shelter was also observed (F = 12.40, p < 0.01), with burrowing behavior under the CWL-S treatment significantly lower than that under both the CWL-NS and RWLR-S treatments (p < 0.05). Similarly, the activity level of P. clarkii was significantly influenced by both water level (F = 22.49, p < 0.001) and shelter (F = 10.13, p < 0.01), with post hoc tests indicating reduced activity under the CWL treatment (p < 0.001) and in the presence of shelter (p < 0.01). A significant interaction between water level and shelter on activity level was also detected (F = 6.40, p = 0.022, Tab. S1, Fig. 1). Previous field studies have indicated that decreasing water levels elevate the activity of P. clarkii. In contrast, rising water levels result in a decrease in activity (Gherardi et al., 2002b). When free water rapidly depletes in the environment, P. clarkii responds to stress by seeking moisture-rich substrates for refuge and initiating burrowing earlier to reduce environmental risks (Correia and Ferreira, 1995; Souty-Grosset et al., 2014). In addition to burrows, P. clarkii utilizes natural refuges for shelter, and the presence of these natural shelters appears to reduce burrowing activity during the warmer season (Ilhéu et al., 2003). However, when faced with extreme cold or substantial water level changes, shelters alone may not be sufficient to meet the survival needs of P. clarkii. As a result, crayfish continue to burrow in search of more secure habitats. In particular, burrows appear to serve as more effective shelters (Ilhéu et al., 2003).

Overall, our results demonstrated that P. clarkii is capable of rapidly burrowing to mitigate the stress of low temperature environments when free water decreases rapidly, regardless of whether it is autumn or winter. This behavior allows P. clarkii to quickly establish microhabitats even during sudden water-level drops in cold winters, thereby preventing freezing mortality under desiccating conditions. Current research indicates that low temperatures do not inhibit the burrowing behavior of P. clarkii. Given its ability to survive the cold season in habitats with seasonal water-level fluctuations, the potential threat posed by P. clarkii to river-connected lakes and floodplain wetland ecosystems is likely to be more substantial.

Acknowledgments

The authors would like to express their gratitude to EditSprings (https://www.editsprings.cn) for the expert linguistic services provided. This study was supported by the National Natural Science Foundation of China (Grant No. 32471648; 32170383) and the open project funding of Key Laboratory of Intelligent Health Perception and Ecological Restoration of Rivers and Lakes, Ministry of Education, Hubei University of Technology (HGKFZ08, HGKFZ04, HGKFZP009).

Supplementary Material

Table. S1. Results of aligned rank transform (ART) factorial ANOVA for behavioral variables of P. clarkii under different water levels, shelter conditions, and seasonal factors.

Video. S1. Five behaviors of Procambarus clarkii.

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References

  • Alcorlo P, Geiger W, Otero M. 2004. Feeding preferences and food selection of the red swamp crayfish, Procambarus clarkii, in habitats differing in food item diversity. Crustaceana 77: 435–453. [Google Scholar]
  • Alcorlo P, Geiger W, Otero M. 2008. Reproductive biology and life cycle of the invasive crayfish Procambarus clarkii (Crustacea: Decapoda) in diverse aquatic habitats of South-Western Spain: Implications for population control. Fundam Appl Limnol 173: 197–212. [Google Scholar]
  • Barbaresi S, Tricarico E, Gherardi F. 2004. Factors inducing the intense burrowing activity of the red-swamp crayfish, Procambarus clarkii, an invasive species. Naturwissenschaften 91: 342–345. [Google Scholar]
  • Barnes N. 2024. Thermal tolerance and burrowing behaviors of red swamp crayfish (Procambarus clarkii). Auburn University. [Google Scholar]
  • Cai FJ, Wu ZJ, He N, Ning L, Huang CM. 2010. Research progress in invasion ecology of Procambarus clarkia. Chin J Ecol 29: 124–132. [Google Scholar]
  • Chucholl C. 2011. Population ecology of an alien “warm water” crayfish (Procambarus clarkii) in a new cold habitat. Knowl Managt Aquatic Ecosyst: 29. [Google Scholar]
  • Chung YS, Cooper RM, Graff J, Cooper RL. 2012. The acute and chronic effect of low temperature on survival, heart rate and neural function in crayfish (Procambarus clarkii) and prawn (Macrobrachium rosenbergii) species. Open J Mol Integr Physiol 2: 75–86. [Google Scholar]
  • Correia AM, Ferreira O. 1995. Burrowing behavior of the introduced red swamp crayfish Procambarus clarkii (Decapoda: Cambaridae) in Portugal. J Crustacean Biol 15: 248–257. [Google Scholar]
  • Davis K, Huber R. 2007. Activity patterns, behavioural repertoires, and agonistic interactions of crayfish: a non-manipulative field study. Behaviour 144: 229–247. [Google Scholar]
  • Dong FY, Xie WX, Xie S, Liang YG, Huang DM, Chen WX, Hu CL. 2008. The preliminary studies on ecological characters of the cave of Procambarus clarkii (Girard) and its impacts on the security of hydro-constructions. Acta Hydrobiologica Sinica 32: 952–954. [Google Scholar]
  • Foster J, Harper D. 2006. Status of the alien Louisianan red swamp crayfish Procambarus clarkii Girard and the native African freshwater crab Potamonautes loveni in rivers of the Lake Naivasha catchment, Kenya. Freshw Crayfish 15: 189–194. [Google Scholar]
  • Geiger W, Alcorlo P, Baltanas A, Montes C. 2005. Impact of an introduced Crustacean on the trophic webs of Mediterranean wetlands. Biol Invasions 7: 49–73. [Google Scholar]
  • Gherardi F, Acquistapace P, Tricarico E, Barbaresi S. 2002a. Ranging behaviour of the red swamp crayfish in an invaded habitat: the onset of hibernation. Freshw Crayfish 14: 330–337. [Google Scholar]
  • Gherardi F, Tricarico E, Ilhéu M. 2002b. Movement patterns of an invasive crayfish, Procambarus clarkii, in a temporary stream of southern Portugal. Ethol Ecol Evol 14: 183–197. [Google Scholar]
  • Gherardi F. 2006. Crayfish invading Europe: the case study of Procambarus clarkii. Mar Freshw Behav Physiol 39: 175–191. [Google Scholar]
  • Giovani SF, Felipe M, Ekaterina B, Rodrigo MA, Armen RK. 2019. Making soil particle size analysis by laser diffraction compatible with standard soil texture determination methods. Soil Sci Soc Am J 83: 1244–1252. [Google Scholar]
  • Gong SY, Lv JL, Sun RP, Li LP, He XG. 2008. The study on reproductive biology of Procambarus clarkii. Freshw Fisheries 6: 23–25. [Google Scholar]
  • Haubrock PJ, Inghilesi AF, Mazza G, Bendoni M, Solari L, Tricarico E. 2019a. Burrowing activity of Procambarus clarkii on levees: analysing behaviour and burrow structure. Wetl Ecol Manag 27: 497–511. [Google Scholar]
  • Haubrock PJ, Kubec J, Veselý L, Buřič M, Tricarico E, Kouba A. 2019b. Water temperature as a hindrance, but not limiting factor for the survival of warm water invasive crayfish introduced in cold periods. J Great Lakes Res 45: 788–794. [Google Scholar]
  • Herrmann A, Martens A. 2024. Burrowing and soil dependence in the invasive crayfish Faxonius immunis under simulated drought conditions. Knowl Managt Aquatic Ecosyst: 22. [Google Scholar]
  • Ilhéu M, Acquistapace P, Benvenuto C, Gherardi F. 2003. Shelter use of the Red Swamp Crayfish (Procambarus clarkii) in dry-season stream pools. Archiv für Hydrobiol 57: 535–546. [Google Scholar]
  • Kingsford RT. 2000. Ecological impacts of dams, water diversions and river management on floodplain wetlands in Australia. Austral Ecol 25: 109–127. [Google Scholar]
  • Kouba A, Tíkal J, Císař P, Veselý L, Fořt M, Příborský J, Patoka J, Buřič M. 2016. The significance of droughts for hyporheic dwellers: evidence from freshwater crayfish. Sci Rep 6: 26569. [CrossRef] [PubMed] [Google Scholar]
  • Kuklina I, Altintas BY, Císař P, Kozák P, Buřič M. 2022. Cardiac activity with acute exposure to sub-zero temperatures illustrates the survival mechanism of invasive crayfish. Limnologica 93: 125962. [Google Scholar]
  • Lozán JL. 2000. On the threat to the European crayfish: a contribution with the study of the activity behaviour of four crayfish species (Decapoda: Astacidae). Limnologica 30: 156–161. [Google Scholar]
  • Oficialdegui FJ, Sánchez MI, Clavero M. 2020. One century away from home: how the red swamp crayfish took over the world. Rev Fish Biol Fisheries 30: 121–135. [Google Scholar]
  • Peeters ET, de Vries R, Elzinga J, Ludányi M, van Himbeeck R, Roessink I. 2024. Triggers affecting crayfish burrowing behaviour. Aquat Ecol 58: 191–206. [CrossRef] [Google Scholar]
  • Souty-Grosset C, Reynolds J, Gherardi F, Aquiloni L, Coignet A, Pinet F, Mancha Cisneros MDM. 2014. Burrowing activity of the invasive red swamp crayfish, Procambarus clarkii, in fishponds of La Brenne (France). Ethol Ecol Evol 26: 263–276. [CrossRef] [Google Scholar]
  • Veselý L, Buřič M, Kouba A. 2015. Hardy exotics species in temperate zone: can “warm water” crayfish invaders establish regardless of low temperatures? Sci Rep 5: 16340. [CrossRef] [PubMed] [Google Scholar]
  • Wobbrock JO, Findlater L, Gergle D, Higgins JJ. 2011. The aligned rank transform for nonparametric factorial analyses using only anova procedures, in: Proceedings of the SIGCHI Conference on Human Factors in Computing Systems. pp. 143–146. [Google Scholar]
  • Zha LL, Xu ZH, Zhang Y. 2022. Effects of different pretreatment methods on grain size characteristics of sediments from a landslide-dammed lake based on mastersizer 2000. Quat Sci 42: 1643–1654. [Google Scholar]
  • Zhang GX, Wu Y, Wu YF, Liu XM. 2018. A review of research on wetland ecohydrology. Adv Water Sci 29: 737–749. [Google Scholar]

Cite this article as: Gao J, Guo Z, Chi Y, Zhang H, Liao M, Wang H. 2025. Cold-season challenges: Behavioral adaptations of invasive Procambarus clarkii to rapid water level decline and shelter availability. Knowl. Manag. Aquat. Ecosyst., 426. 25. https://doi.org/10.1051/kmae/2025020

All Tables

Table 1

Burrowing and activity performance (Mean ± SD) of P. clarkii under different water level, shelter, and season treatments. RWLR-NS indicates Rapid Water Level Reduction without Shelter; CWL-NS, Constant Water Level without Shelter; RWLR-S, Rapid Water Level Reduction with Shelter; CWL-S, Constant Water Level with Shelter, n = 3 tanks per treatment.

In the text

All Figures

thumbnail Fig. 1

Mean (± SE) percentages of behavioral of P. clarkii under different combinations of water level, shelter availability, and season. The six panels represent: (1) movement, (2) fighting, (3) resting, (4) burrowing, (5) sideways breathing, and (6) overall activity level. RWLR-NS indicates Rapid Water Level Reduction without Shelter; CWL-NS, Constant Water Level without Shelter; RWLR-S, Rapid Water Level Reduction with Shelter; CWL-S, Constant Water Level with Shelter, n = 3 tanks per treatment.
In the text

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