Living on the edge: Crayfish as drivers to anoxification of their own shelter microenvironment.

Burrowing is a common trait among crayfish thought to help species deal with adverse environmental challenges. However, little is known about the microhabitat ecology of crayfish taxa in relation to their burrows. To fill this knowledge gap, we assessed the availability of oxygen inside the crayfish shelter by series of in-vivo and in-silico modelling experiments. Under modeled condition, we found that, except for the entrance region of the 200 mm, a flooded burrow microenvironment became anoxic within 8 h, on average. Multiple 12-hour day-night cycles, with burrows occupied by crayfish for 12 h and empty for 12 h, were not sufficient for refreshing the burrow microenvironment. We then examined the degree to which crayfish species with different propensities for burrowing are tolerant of self-created anoxia. From these experiments, primary and secondary burrowers showed best and most consistent tolerance-exhibiting ≥ 64% survival to anoxia and 25-91% survival of ≥ 9 h at anoxia, respectively. Tertiary burrowers exhibited little to no tolerance of anoxia with 0-50% survival to anoxia and only one species exhibiting survival (2%) of ≥ 9 h at anoxia. Results suggest that moderate to strongly burrowing crayfish can quickly draw down the dissolved oxygen in burrow water but appear to have conserved a legacy of strong tolerance of anoxia from their monophyletic ancestors-the lobsters-whereas tertiary burrowers have lost (or never evolved) this ability.

Effect of psychoactive substances on cardiac and locomotory activity of juvenile marbled crayfish Procambarus virginalis.

Pharmaceutically active compounds are common and increasing in the aquatic environment. Evidence suggests they have adverse effects on non-target organisms, and they are classified as emerging pollutants for a variety of aquatic organisms. To determine the effects of environmentally relevant levels of psychoactive compounds on non-target organisms, we analyzed cardiac and locomotory activity in early developmental stages of marbled crayfish Procambarus virginalis. Responses to sertraline, methamphetamine, and a mixture of citalopram, oxazepam, sertraline, tramadol, venlafaxine, and methamphetamine at a concentration of 1 µg L-1 of each compound were assessed. On day four of exposure, cardiac activity was recorded for 5 min, and on day eight, locomotory activity was recorded for 15 min. There was a significant increase (p < 0.01) in heart rate in methamphetamine-exposed and Mix-exposed juveniles compared to the unexposed control and there was significant difference (p < 0.01) in proportion of time (activity %) was observed with sertraline-exposed, whereas velocity, and distance moved did not significantly differ (p > 0.05) in exposed and control animals. These findings revealed that low concentrations of chemicals and their mixtures can modify the physiological state of aquatic animals without outward manifestations (activity, distance moved, and velocity). Aquatic animals can be impacted earlier than is visible, but effects can potentially lead to substantial changes in populations and in ecosystem processes. Additional research to investigate chemical combinations, exposure systems, and organism physiological and molecular responses may provide evidence of broad impact of environmental pharmaceuticals.

Short-term effects of an environmentally relevant concentration of organic UV filters on signal crayfish Pacifastacus leniusculus.

Personal care products, including organic UV filters, are considered emerging contaminants, with their toxic effects being a concern in recent decades. UV filters continually enter surface waters via wastewater and human activity. Despite the presence of organic UV filters in the freshwater environment, little is known of their impact on aquatic biota. In this study, we evaluated the cardiac and locomotor responses of signal crayfish Pacifastacus leniusculus exposed to environmentally relevant concentrations of either 2-Phenylbenzimidazole-5-sulfonic acid (PBSA, 3 µg/L) or 5-Benzoyl-4-hydroxy-2-methoxybenzenesulfonic acid (BP4, 2.5 µg/L). Specimens exposed to the tested compounds for 30 min exhibited significantly greater changes in distance moved and time active than did unexposed controls. Significant differences of mean heart rate change compared to control were detected in both PBSA and BP4 experimental groups. Such behavior and physiological alterations demonstrate ecological effects of personal care products with the tested sunscreen compounds even with a short exposure. Evidence of the consequences of organic UV filters on aquatic organisms is scarce and is an important topic for future research.

Responses of signal crayfish Pacifastacus leniusculus to single short-term pulse exposure of pesticides at environmentally relevant concentrations.

Although pesticides are often discharged into surface waters in pulses as opposed to a sustained release, the effect of episodic pollution events on freshwater crayfish is largely unknown. We monitored change in heart rate and distance moved to assess the response of signal crayfish Pacifastacus leniusculus to short-term exposure to environmentally relevant concentrations of metazachlor (MTZ), terbuthylazine (TER), and thiacloprid (TCL). Crayfish exposed to 20 µg/L of MTZ exhibited a significant increase in mean heart rate and distance moved. Increased heart rate was detected at 118 ± 74 s post-exposure to MTZ. There were no significant differences in mean heart rate and distance moved in crayfish exposed to 6 µg/L of TCL and 4 µg/L of TER. A significant correlation between heart rate and distance moved was found in all exposed groups. These results suggest that pulse exposure to MTZ impact crayfish physiology and behavior during short-term period. With pulse exposure to TCL and TER, crayfish not exhibiting a locomotor response may continue to be exposed to lower, but potentially harmful, levels of pollutants. Evidence of the impacts of pesticide pulse at environmentally relevant concentrations on crayfish is scarce. Further study is required to determine the ecological effects of such events on freshwater crayfish.

Cardiac and Locomotor Responses to Acute Stress in Signal Crayfish Pacifastacus leniusculus Exposed to Methamphetamine at an Environmentally Relevant Concentration.

Methamphetamine (METH), a central nervous system stimulant used as a recreational drug, is frequently found in surface waters at potentially harmful concentrations. To determine effects of long-term exposure to environmentally relevant levels on nontarget organisms, we analysed cardiac and locomotor responses of signal crayfish Pacifastacus leniusculus to acute stress during a 21-day exposure to METH at 1 µg L-1 followed by 14 days depuration. Heart rate and locomotion were recorded over a period of 30 min before and 30 min after exposure to haemolymph of an injured conspecific four times during METH exposure and four times during the depuration phase. Methamphetamine-exposed crayfish showed a weaker cardiac response to stress than was observed in controls during both exposure and depuration phases. Similarly, methamphetamine-exposed crayfish, during METH exposure, showed lower locomotor reaction poststressor application in contrast to controls. Results indicate biological alterations in crayfish exposed to METH at low concentration level, potentially resulting in a shift in interactions among organisms in natural environment.

Continuous Noninvasive Measuring of Crayfish Cardiac and Behavioral Activities.

A crayfish is a pivotal aquatic organism that serves both as a practical biological model for behavioral and physiological studies of invertebrates and as a useful biological indicator of water quality. Even though crayfish cannot directly specify the substances that cause water quality deterioration, they can immediately (within a few seconds) warn humans of water quality deterioration via acute changes in their cardiac and behavioral activities. In this study, we present a noninvasive method that is simple enough to be implemented under various conditions due to a combination of simplicity and reliability in one model. This approach, in which the biological organisms are implemented into environmental evaluation processes, provides a reliable and timely alarm for warning of and preventing acute water deterioration in an ambient environment. Therefore, this noninvasive system based on crayfish physiological and ethological parameter recordings was investigated for the detection of changes in an aquatic environment. This system is now applied at a local brewery for controlling quality of the water used for beverage production, but it can be used at any water treatment and supply facility for continuous, real-time water quality evaluation and for regular laboratory investigations of crayfish cardiac physiology and behavior.

Crayfish can distinguish between natural and chemical stimuli as assessed by cardiac and locomotor reactions.

In this study, cardiac and locomotor activities of signal crayfish Pacifastacus leniusculus were investigated under exposure to a range of natural (i.e., odors of conspecific crayfish, predatory fish, food, and injured conspecific) and one chemical (i.e., disinfectant chloramine-T) stimuli. Crayfish locomotion was simultaneously initiated with an increase in heart rate only when affected by chloramine-T, while locomotor response was delayed in all cases (or was not manifested at all by some specimens) when disturbed by the natural stressors. The heart rate differences measured before and during the stimulation were arranged as follows: odor of conspecific crayfish (9.2 ± 7.1%) < predator (18.4 ± 13%) < food (33.5 ± 15.7%) < chloramine-T (41.1 ± 14.7%) < injured conspecific (51.8 ± 28.4%). Analysis of the peculiarities of crayfish heartbeat under exposure to the tested stimuli revealed complex cardiac responses as was previously observed by an electrocardiography approach, that is, a slowed heart rate followed by a delayed increase. Evaluation of the intrinsic parameters of crayfish bioindicators remains essential due to the possibility of detection of the substantial ethological responses even in motionless animals. The role and appropriateness of signal crayfish as a bioindicator of water quality is discussed; they seem to be an applicable species for this task due to their sufficient sensitivity and broad availability. In addition to providing a better understanding of stereotypic crayfish behaviors induced by common and chemical stressors, the results of this study may serve as reference data for the evaluation of crayfish suitability for water quality tests.

Investigation of chloramine-T impact on crayfish Astacus leptodactylus (Esch., 1823) cardiac activity.

The crayfish play an essential role in the biomonitoring and may reflect ambient water quality through the biochemical, behavioural and physiological reactions. To assess whether narrow-clawed crayfish Astacus leptodactylus can respond by heart rate changes to presence in water of such biocide as chloramine-T, adult males were exposed to its low (2 and 5 mg L(-1)), moderate (10 mg L(-1), commonly used in industry and aquaculture) and exceeded (20 and 50 mg L(-1)) concentrations. In addition, a physical stress test evaluated energy expenditure following the chemical trials. Three key reactions (cardiac initial, first-hour and daily prolonged exposure) were discussed with particular focus on crayfish initial reaction as the most meaningful in on-line water quality biomonitoring. After short-term exposure to both chloramine-T concentrations, crayfish were found to respond rapidly, within 2-5 min. According to heart rate changes, the 1-h exposure did not adversely affect crayfish at either concentration, as well as during daily exposure to 10 mg L(-1). As assessed by the heart rate, the 24-h exposure to 50 mg L(-1) of chloramine-T was toxic for crayfish and led to substantial loss of energy that became apparent during subsequently conducted physical stress. The results supported a hypothesis that crayfish vital functions are connected with environment they inhabit closely enough to serve as biological monitors. Crayfish were tolerant to short-term chloramine-T exposure, while rapid crayfish reaction to an increased chemical level indicated their high sensitivity, an essential attribute of real-time environmental assessment.

Accumulation of heavy metals in crayfish and fish from selected Czech reservoirs.

To evaluate the accumulation of aluminium, cadmium, chromium, copper, lead, mercury, nickel, and zinc in crayfish and fish organ tissues, specimens from three drinking water reservoirs (Boskovice, Landstejn, and Nová Ríse) and one contaminated site (Darkovské more) in the Czech Republic were examined. Crayfish hepatopancreas was confirmed to be the primary accumulating site for the majority of metals (Cu > Zn > Ni > Cd > Cr), while Hg and Cr were concentrated in abdominal muscle, and Al and Pb were concentrated in gill. Metals found in Nová Ríse specimens included Cu > Zn > Ni and those found in Boskovice included Zn > Hg > Cr. Cd concentrations were observed only in Landstejn specimens, while contaminated Darkovské more specimens showed the highest levels of accumulation (Cu > Al > Zn > Pb). The majority of evaluated metals were found in higher concentrations in crayfish: Cu > Al > Zn > Ni > Cr > Cd > Pb, with Hg being the only metal accumulating higher in fish. Due to accumulation similarities of Al in crayfish and fish gill, differences of Hg in muscle, and features noted for the remaining metals in examined tissues, biomonitoring should incorporate both crayfish and fish to produce more relevant water quality surveys.

Real-time monitoring of water quality using fish and crayfish as bio-indicators: a review.

Water quality monitoring using fish and crayfish as bio-indicators requires an understanding of the state of pollution of waters, choice of bio-indicators, physiological and behavioral endpoints of fish and crayfish, and principles of the methodology and their potential applications. Here, we discuss telemetry, acoustic monitoring, vision-based monitoring, measures of ventilatory activity, electrocardiography, and fiber-optic plethysmography. Assessment of water quality must be based, not only on physicochemical characteristics of the current environment as determined by chemical analyses, but also on observations of the physiology and behavior of its inhabitants. Real-time biomonitoring is suggested as the most reliable method, since it incorporates living organisms into the system to serve as biosensors. The potential application of the methods discussed includes use at water treatment plants and water supply stations for prevention of hazardous toxicological events, and, for aquaculture, in ponds, lakes, and aquariums for monitoring growth, population size, and behavior traits.