Next Article in Journal
Analysis of Flood Fatalities–Slovenian Illustration
Previous Article in Journal
Impact of Bed Form Celerity on Oxygen Dynamics in the Hyporheic Zone
 
 
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Crayfish as Bioindicators for Monitoring ClO2: A Case Study from a Brewery Water Treatment Facility

Research Institute of Fish Culture and Hydrobiology, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Faculty of Fisheries and Protection of Waters, University of South Bohemia in České Budejovice, Zátiší 728/II, 38925 Vodňany, Czech Republic
*
Author to whom correspondence should be addressed.
Water 2020, 12(1), 63; https://doi.org/10.3390/w12010063
Received: 28 November 2019 / Revised: 17 December 2019 / Accepted: 20 December 2019 / Published: 23 December 2019
(This article belongs to the Section Wastewater Treatment and Reuse)

Abstract

:
This study focuses on the use of crayfish as bioindicators in the water treatment process during operating conditions. The crayfish physiological responses to water disinfected with chlorine dioxide (ClO2) was evaluated. Monitoring was conducted at the private commercial enterprise Protivín Brewery in Czech Republic under standard operating conditions. This brewery has a water treatment facility, where ClO2 is used for water purification. A total of 25 adult signal crayfish (Pacifastacus leniusculus) were kept in separate flow-through aquaria receiving the purified water with ClO2 concentrations ranging from 0.01 to 0.29 mg L−1. Diurnal rhythms of 32% of crayfish was disturbed even at lower concentrations of ClO2 (0.01–0.2 mg L−1), while higher concentrations (>0.2 mg L−1) affected all animals. A random decline and rise of heart rate was detected. In addition, the frequent occurrence of higher levels of ClO2 significantly increased mortality. On average, mortality of crayfish occurred three to four weeks after stocking into the experimental system. Crayfish mortality is estimated to occur at concentrations exceeding 0.2 mg L−1 of ClO2. Our results suggest that long-term exposure to ClO2 adversely affects crayfish physiology. In addition, the results of this study could contribute to the use of crayfish as bioindicators in long-term water quality monitoring under industrial conditions.

1. Introduction

Decapods, such as crayfish, are known to be sensitive to contamination in freshwater bodies. Given their sensitivity to changes in water quality, these organisms are highly responsive to changes in aquatic ecosystems [1,2,3]. Crayfish have been used as bioindicators both in the aquatic environment and under laboratory conditions. They demonstrate an affinity for accumulating pollutants in their tissues [1,4,5], and elicit a response to different substances [2,3,6]. Subsequently, there is a potential for their use as bioindicators in practical monitoring under industrial conditions.
Given that crayfish are nocturnal, their heart rate and locomotor activity increase at night [7,8,9]. However, the crayfish heart rate can also be influenced by certain stimuli [7,8,9]. Several studies have described the crayfish cardiac response to chemical stimuli, including chlorine organic compounds [3,10] and chloride content in water [11].
While different compounds may be used for water purification, the most effective disinfectant is chlorine dioxide [12]. It is a powerful oxidant among chlorine compounds and it is widely applied in surface water disinfection [13]. There are a few chemical reactions that produce chlorine dioxide (ClO2), and one of them is the hydrochloric-acid-sodium-chlorite reaction [13]:
5NaClO2 + 4HCl = 4ClO2 + 5NaCl + 2H2O.
Generally, ClO2 treatment concentrations may range from 0.07 to 2 mg L−1, which is sufficient for water disinfection [14]. Chlorine dioxide can be effectively applied for iron and manganese oxidation at temperatures as low as 2 °C and a pH of 5.5 [15]. Moreover, ClO2 is efficient at removing both tastes and odors [12], and its threshold in this case could be as low as 0.2 mg L−1 [14]. During water treatment ClO2 is reduced to its main decomposition product, chlorite (ClO2) [13,16]. Subsequently, the levels of ClO2 are directly dependent on the concentration of ClO2 used. Hence, it is important to maintain ClO2 levels during water treatment, in order to prevent chlorite levels exceeding the WHO guideline value [14].
Given that ClO2 is a widely used disinfectant, it is important to understand its effect on living organisms. Currently, the effects of ClO2 on aquatic organisms remain poorly described and mainly focus on teleost fish [17,18,19]. Further, one study describes ClO2 toxicity to zebra mussel Dreissena polymorpha [20].
The present study investigated the efficacy of crayfish as bioindicators for monitoring ClO2 levels during the water treatment process employed by a local brewery, focusing on the biological response and lethal concentration of adult signal crayfish Pacifastacus leniusculus to long term ClO2 exposure.

2. Materials and Methods

2.1. Monitoring Process

Monitoring was conducted from February to August 2017 under the running conditions of the private enterprise, Protivín Brewery, Protivín, Czech Republic. This practical investigation was operational with crayfish since April 2016 and data tracking commenced from February 2017. The brewery has a water-treatment facility, where ClO2 is used for water purification. ClO2 was produced by the hydrochloric-acid-sodium-chlorite reaction. In this reaction ClO2 yields and conversion had different values, where maximum yield is 100% and maximum conversion is 80%, which is sufficient for water treatment [13]. Water ClO2 concentrations were measured daily. All crayfish were exposed to ClO2 during monitoring. Due to the operating conditions of the enterprise, the use of an uncontaminated control group was not possible. However, previous studies have clearly described the typical dynamics of the heart rate of crayfish [8,21].

2.2. Monitoring System

This study made use of the noninvasive crayfish cardiac activity monitoring (NICCAM) system described by Pautsina et al. [22]. This NICCAM system consists of a multichannel 14 bit analog-to-digital converter (ADC) with USB interface, personal computer with software for data processing and infrared (IR) optical sensors.
This system could monitor, record, and analyze crayfish cardiac activity, expressed as heart rate, and store the text files digitally. The software graphical user interface displayed raw cardiac activity signals of five crayfish simultaneously.
The sensors were fixed with non-toxic two-component epoxy adhesive on the dorsal side of each crayfish carapace above the heart at the locality where the strongest heart rate was detected. Glue hardened in approximately 15 min. The attached sensor still allowed crayfish to move freely around the aquarium. The monitored cardiac activity signals of the crayfish were recorded and displayed on the software’s graphical user interface in real-time. Data about cardiac activity were continuously logged onto a personal computer and then processed using MS Excel for further analyzing based on created diagrams. Given that a single crayfish successful molted during the monitoring period, its pre-ecdysis period was also analyzed.

2.3. Experimental Animals

Adult signal crayfish P. leniusculus were obtained from ponds near Velké Meziříčí (49.3788544 N, 16.0825961 E) in the Vysočina Region, Czech Republic. Non-native crayfish species were used given the protected status of indigenous species and the regulations against their manipulation. The present study was carried out under the practical running conditions of the brewery, which mitigated risks associated with escape and species introduction and permitted the use of the non-native crayfish.
Before commencing the experiment, crayfish were acclimated for two weeks to the laboratory conditions of the Faculty of Fisheries and Protection of Waters, University of South Bohemia in České Budějovice, Vodňany, Czech Republic. Crayfish were individually kept in recirculating aquarium systems. Feeding and water changes were provided twice per week. No mortality was observed during the acclimation period.
Before monitoring commenced, the crayfish (with attached sensors) were acclimatized to lower temperatures in incubators (thermostatic cabinets Liebherr FK 5440, Liebherr-Hausgeräte Ochsenhausen GmbH, Ochsenhausen, Germany), where the temperature was decreased by 1 °C each following day. When acclimated temperature reached 10 °C, crayfish were transported in thermo-boxes in a small amount of water from the laboratory to the brewery by car (approximately 10 km). Before the experiment, crayfish were visually examined for absence of diseases and their biometrical measurements collected: Carapace length (mean ± SD): 43.8 ± 0.77 mm; total length: 90.13 ± 1.6 mm; and total weight: 33.68 ± 2.03 g. Weight and length was measured with digital calipers (Schut Geometrical Metrology, Groningen, The Netherlands) and an electronic balance (Kern & Sohn GmbH, Balingen, Germany). Both sexes of crayfish were used based on the previous study [23] which found no substantial differences between their reactions to stimuli. Only crayfish with intact appendages (antennae, chelae, and walking legs) were used in the experiment. During the experiment, crayfish were kept in separate 10 L flow-through aquariums, each receiving ClO2-treated water with temperature of 10 ± 0.5 °C and pH 8.3 ± 0.5, under constant photoperiod 12:12 light-dark cycle. Each aquarium was provided with an artificial shelter (halved ceramic flower pot). A hole made on the upper surface of the shelter permitted recording of cardiac activity, even when crayfish (with the attached sensors) were inside the shelter. The experimental system could hold ten crayfish simultaneously. Five crayfish received a heart rate monitor each, while the other five crayfish were kept as reserves. In case of molting or mortality, an individual was replaced by one of the reserves. Thus, twenty-five animals were used in total throughout the monitoring period. Animals were fed daily with commercial food pellets (Sera GmbH; Heinsberg, Germany), and remains and feces were removed via daily syphoning.

2.4. Statistical Analysis

The data recorded from treated crayfish was divided between three groups in accordance with the day of exposure to maximum ClO2 concentration (Cmax; ClO2 > 0.2 mg L−1): Group one got Cmax on day 4 ± 2; Group two on day 13 ± 1; and Group three was exposed to Cmax on day 38 ± 6 after stocking to experimental aquarium system (Table 1). The data was grouped for subsequent analysis.
Shapiro-Wilk’s test was used to assess the normality of residuals. Data were transformed when necessary to meet the assumptions of normality and equal variance. Differences in life duration after ClO2 Cmax exposure between tested groups were estimated by one-way analysis of variance (ANOVA) and subsequent post hoc Tukey’s test (Statistica 13, StatSoft, Inc., Tulsa, OK, USA). Data are presented as means ± standard deviation (SD). The level of significance was set at p < 0.05.

3. Results

3.1. Ecdysis Period

While five unsuccessful moltings resulted in crayfish mortality, a single molting proved successful. The highest heart rate was recorded 4 h before the molting, ranging between 39 and 60 beats per minute (bpm), with a peak of 72 bpm (Figure 1). Heart rate declined 35 min before molting with a few “leaps”.

3.2. Diurnal Rhythm

Crayfish were exposed to ClO2 concentrations ranging from 0.01 to 0.29 mg L−1. These concentrations varied every day (Figure 2). Following monitoring, the data was divided according to the number of high concentrations of ClO2. During the first three months (February–April), high ClO2 (0.2–0.29 mg L−1) concentrations were recorded 4.6 times less than during the next four months (May–August), when high concentrations occurred more often.
The heart rate daily cycle of 32% of crayfish was already disturbed at a lower level of ClO2 concentration (less than 0.2 mg L−1). A prevalence of disrupted heart rate was observed, with chaotic increases and decreases regardless of the time of day. There was no statistical difference between day and night cardiac activities within the tested groups (Table 2) as well as between groups (day: F(2,22) = 0.80780, p = 0.45863 and night: F(2,22) = 1.5974, p = 0.22503). The diurnal rhythm was disrupted, and cardiac rhythmicity was lost. This was expressed in different heart rate fluctuations of animals exposed to the same concentrations of ClO2 (Figure 3).

3.3. Mortality

High ClO2 concentrations (0.2–0.29 mg L−1) disturbed the diurnal rhythm of all individuals, inducing loss of rhythmicity and subsequent mortality (Figure 4). Mortality increased along with more frequent occurrences of high ClO2 concentrations. During the first period (89 days), where high ClO2 concentrations (higher than 0.2 mg L−1) were recorded five times, four crayfish died. During the second 113-day period, where high ClO2 concentrations occurred 23 times, 21 crayfish died. Thus, in the first period mortalities occurred 5.3 times less than in the second one. No individual survived the experiment (Figure 4).

Life Duration after Exposure to Cmax

Life duration after exposure to Cmax for each crayfish was determined (Table 1). There was a significant difference (p < 0.05) in life duration between groups. Crayfish from Group two generally lived twice as long (16 ± 8 days) as crayfish from Groups one and three (9 ± 7 and 5 ± 2 days, respectively) after exposure to Cmax (Figure 5).

4. Discussion

In the present study, the effect of long-term exposure of signal crayfish to different levels of ClO2 has been investigated and assessed through the observation of heart rate, diurnal rhythm and mortality.
A single recorded molting was preceded by rapid heart rate fluctuations. The increase in heart rate was observed four hours before the molting, up to 60 bpm with the peak of 72 bpm, and the heart rate decline was detected 35 min before molting (Figure 1). Kuramoto [24] described the cardiac changes of untreated spiny lobster Panulirus japonicus before the molting and noted that the heart rate rose and fell during molting of lobster similarly to that of crayfish. The heart rate of an unaffected lobster increased 1–2 h before ecdysis to a peak of 80–120 bpm and declined about 15 min before the beginning of molting. Thus, the changes in the heart rate of unstimulated spiny lobster and the ClO2 exposed signal crayfish were similar in the premolting period.
Unsuccessful molting was also observed to result in death. In Kuklina et al. [3], chloramine-T exposed narrow-clawed crayfish Astacus leptodactylus, suffered from lack of energy when exposed to physical stress. Energetic deficiency can be a potential reason for unsuccessful molting in our study, where ClO2 exposure depleted crayfish energy stores and prohibited molting, resulting in their mortality.
Owing to their nocturnal nature, the narrow-clawed crayfish A. leptodactylus heart rate is higher at night than during the day, even at temperatures below 14 °C [8]. The present study showed an impact of ClO2 on crayfish heart rate and nocturnal behavior. A disturbance of the circadian cardiac rhythm was observed in all individuals, expressed as a random decline and rise of heart rate, regardless of the time of day. The typical increased nocturnal heart rate was not noticed at the lowest ClO2 concentration in 32% of the crayfish, while in the high concentrations it was completely disrupted for all animals. As soon as the diurnal rhythm was disturbed, the circadian rhythmicity was lost, demonstrating impaired cardiac function and leading to crayfish mortality (Figure 3). A similar observation was described in Kuznetsova et al. [21] where highly concentrated hydroquinone solution (1 g L−1) disrupted A. leptodactylus circadian rhythm before death. Styrishave et al. [7] noticed that heart rate increased during the day and decreased at night in noble crayfish Astacus astacus when exposed to copper (8.0 mg L−1) and mercury (0.1 mg L−1). In this case high mortality (>90%) was detected after 19 days of exposure. Consequently, the ClO2 used in our monitoring and heavy metals used by Styrishave et al. [7] and hydroquinone used in Kuznetsova et al. [21] can be toxic compounds at certain concentrations, and may negatively affect the health of aquatic organisms and even induce their mortality.
Not only are the loss of circadian rhythmicity suspected to induce crayfish mortality, but also the changes in physiology. The gills of fathead minnows Pimephales promelas were negatively affected by 0.13 mg L−1 of ClO2 concentration [25]. Chupani et al. [26] found heavy histopathological changes in crayfish exposed to peracetic acid (2–10 mg L−1), while similar effects often induce mortality in juvenile grass carp Ctenopharyngodon idella [27] and channel catfish Ictalurus punctatus [28]. Subsequently, ClO2 could induce adverse cardio-respiratory responses, reduce larval rainbow trout (Oncorhynchus mykiss) growth in concentration above 0.3 mg L−1 [17] and cause oxidative damage and changes in antioxidant defenses in the heart tissue of juvenile rainbow trout [19]. Hence, it could have a similar effect in crayfish. Moreover, ClO2 is more toxic to aquatic organisms than chlorite and peracetic acid [17,18]. Therefore, considering how ClO2 is harmful for non-target aquatic animals and that it has higher toxicity than other substances, ClO2 might likely have an adverse effect on crayfish tissues, leading to various disorders and subsequent mortality.
Peak concentrations of ClO2 (0.2–0.29 mg L−1) observed during our experiment significantly influenced the life duration of animals. Another study determined that 1–5 mg L−1 of ClO2 induces mortality of zebra mussel D. polymorpha [20].
When the Cmax occurred, crayfish mortality was noticed after approximately 10 ± 7 days. Group one could likely not survive due to immediate exposure to increased ClO2 concentrations, which resulted in rapid mortality. The prolonged exposure of Group three to low-to-medium concentrations of ClO2 resulted in a cumulative effect, preventing organ and tissue regeneration, and resulted in crayfish mortalities 5 ± 2 days after Cmax occurred. Group two, which was exposed to moderate ClO2 concentrations within relatively short time (longer than Group one but shorter than Group three), had the longest life duration after getting Cmax. This may suggest that crayfish responses differ between individuals.

5. Conclusions

Changes in crayfish heart rate and circadian rhythmicity could provide information about their functional state and help us make inferences on environmental state. Crayfish’s physiological sensitivity allow early detection of increased levels of harmful chemicals, thereby presenting a practical solution for proactive water quality monitoring. Our results suggest that the changes in heart rate and diurnal rhythm of treated animals was crayfish-specific, which may stem from their varying functional state and individual physiological response to ClO2 concentrations. There was a direct correlation between Cmax, and crayfish mortality. ClO2 adversely affected crayfish circadian rhythm. In conclusion, this study demonstrated that crayfish could serve as effective bioindicators for long term practical water quality monitoring.

Author Contributions

Conceptualization, V.M., F.L., I.K. and P.K.; Data curation, P.C.; Formal analysis, V.M.; Investigation, V.M., F.L. and I.K.; Methodology, F.L. and I.K.; Project administration, V.M., F.L., I.K. and P.K.; Resources, F.L., I.K. and P.C.; Software, P.C.; Supervision, P.K.; Validation, P.K.; Visualization, V.M. and P.K.; Writing—original draft, V.M. and F.L.; Writing—review & editing, V.M., F.L., I.K., P.C. and P.K. All authors have read and agreed to the published version of the manuscript.

Funding

This study was funded by the Ministry of Education, Youth and Sports of the Czech Republic—project CENAKVA (LM2018099).

Acknowledgments

The authors would like to thank Protivín Brewery, Michal Voldřich and Roman Dědič for their help in conducting the experiment.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Schilderman, P.A.E.L.; Moonen, E.J.C.; Maas, L.M.; Welle, I.; Kleinjans, J.C.S. Use of crayfish in biomonitoring studies of environmental pollution of the river Meuse. Ecotox. Environ. Safe 1999, 44, 241–252. [Google Scholar] [CrossRef] [PubMed][Green Version]
  2. Soedarini, B.; Klaver, L.; Roessink, I.; Widianarko, B.; van Straalen, N.M.; van Gestel, C.A.M. Copper kinetics and internal distribution in the marbled crayfish (Procambarus sp.). Chemosphere 2012, 87, 333–338. [Google Scholar] [CrossRef] [PubMed]
  3. Kuklina, I.; Sladkova, S.; Kouba, A.; Kholodkevich, S.; Kozak, P. Investigation of chloramine-T impact on crayfish Astacus leptodactylus (Esch., 1823) cardiac activity. Environ. Sci. Pollut. Res. 2014, 21, 10262–10269. [Google Scholar] [CrossRef] [PubMed]
  4. Pennuto, C.M.; Lane, O.P.; Evers, D.C.; Taylor, R.J.; Loukmas, J. Mercury in the northern crayfish, Orconectes virilis (Hagen), in New England, USA. Ecotoxicology 2005, 14, 149–162. [Google Scholar] [CrossRef] [PubMed][Green Version]
  5. Alcorlo, P.; Otero, M.; Crehuet, M.; Baltanas, A.; Montes, C. The use of the red swamp crayfish (Procambarus clarkii, Girard) as indicator of the bioavailability of heavy metals in environmental monitoring in the River Guadiamar (SW, Spain). Sci. Total Environ. 2006, 366, 380–390. [Google Scholar] [CrossRef] [PubMed]
  6. Nathaniel, T.I.; Huber, R.; Panksepp, J. Repeated cocaine treatments induce distinct locomotor effects in Crayfish. Brain Res. Bull. 2012, 87, 328–333. [Google Scholar] [CrossRef] [PubMed]
  7. Styrishave, B.; Rasmussen, A.D.; Depledge, M.H. The Influence of Bulk and Trace Metals on the Circadian Rhythm of Heart Rates in Fresh Water Crayfish. Astacus astacus. Mar. Pollut Bull. 1995, 31, 87–92. [Google Scholar] [CrossRef]
  8. Udalova, G.P.; Khodasevich, S.V.; Sladkova, S.V.; Ivanov, A.V.; Rymsha, V.A. Study of circadian activity in the crayfish Pontastacus leptodactylus during their multimonth maintenance in the river water flow. J. Evol. Biochem. Physiol. 2009, 45, 372–381. [Google Scholar] [CrossRef]
  9. Bojsen, B.H.; Witthofft, H.; Styrishave, B.; Andersen, O. In situ studies on heart rate and locomotor activity in the freshwater crayfish, Astacus astacus (L.) in relation to natural fluctuations in temperature and light intensity. Freshw. Biol. 1998, 39, 455–465. [Google Scholar] [CrossRef]
  10. Kholodkevich, S.V.; Ivanov, A.V.; Kurakin, A.S.; Kornienko, E.L.; Fedotov, V.P. Real time biomonitoring of surface water toxicity level at water supply stations. Environ. Bioindic. 2008, 3, 23–34. [Google Scholar] [CrossRef]
  11. Kozak, P.; Policar, T.; Fedotov, V.P.; Kuznetsova, T.V.; Buric, M.; Kholodkevich, S.V. Effect of chloride content in water on heart rate in narrow-clawed crayfish (Astacus leptodactylus). Knowl. Manag. Aquat. Ecosyst. 2009, 394–395, 10. [Google Scholar] [CrossRef][Green Version]
  12. Lalezary, S.; Pirbazari, M.; Mcguire, M.J. Oxidation of 5 Earthy Musty Taste and Odor Compounds. J. Am. Water Works Assoc. 1986, 78, 62–69. [Google Scholar] [CrossRef]
  13. Aieta, E.M.; Berg, J.D. A Review of Chlorine Dioxide in Drinking-Water Treatment. J. Am. Water Works Assoc. 1986, 78, 62–72. [Google Scholar] [CrossRef]
  14. World Health Organization. Chlorine Dioxide, Chlorite and Chlorate in Drinking-water. In Background Document for Development of WHO Guidelines for Drinking-Water Quality; World Health Organization: Geneva, Switzerland, 2016; p. 24. [Google Scholar]
  15. Knocke, W.R.; Vanbenschoten, J.E.; Kearney, M.J.; Soborski, A.W.; Reckhow, D.A. Kinetics of Manganese and Iron Oxidation by Potassium-Permanganate and Chlorine Dioxide. J. Am. Water Works Assoc. 1991, 83, 80–87. [Google Scholar] [CrossRef]
  16. Fisher, D.J.; Burton, D.T.; Yonkos, L.T.; Turley, S.D.; Ziegler, G.P.; Turley, B.S. Derivation of acute ecological risk criteria for chlorite in freshwater ecosystems. Water Res. 2003, 37, 4359–4368. [Google Scholar] [CrossRef]
  17. Svecevicius, G.; Syvokiene, J.; Stasiunaite, P.; Mickeniene, L. Acute and chronic toxicity of chlorine dioxide (ClO2) and chlorite (ClO2-) to rainbow trout (Oncorhynchus mykiss). Environ. Sci. Pollut. Res. 2005, 12, 302–305. [Google Scholar] [CrossRef]
  18. Elia, A.C.; Anastasi, V.; Dorr, A.J.M. Hepatic antioxidant enzymes and total glutathione of Cyprinus carpio exposed to three disinfectants, chlorine dioxide, sodium hypochlorite and peracetic acid, for superficial water potabilization. Chemosphere 2006, 64, 1633–1641. [Google Scholar] [CrossRef]
  19. Tkachenko, H.; Kurhaluk, N.; Grudniewska, J. Biomarkers of oxidative stress and antioxidant defences as indicators of different disinfectants exposure in the heart of rainbow trout (Oncorhynchus mykiss Walbaum). Aquac. Res. 2015, 46, 679–689. [Google Scholar] [CrossRef]
  20. Matisoff, G.; Brooks, G.; Bourland, B.I. Toxicity of chlorine dioxide to adult zebra mussels. J. Am. Water Works Assoc. 1996, 88, 93–106. [Google Scholar] [CrossRef]
  21. Kuznetsova, T.V.; Sladkova, G.V.; Kholodkevich, S.V. Evaluation of functional state of crayfish Pontastacus leptodactylus in normal and toxic environment by characteristics of their cardiac activity and hemolymph biochemical parameters. J. Evol. Biochem. Physiol. 2010, 46, 241–250. [Google Scholar] [CrossRef]
  22. Pautsina, A.; Kuklina, I.; Stys, D.; Cisar, P.; Kozak, P. Noninvasive crayfish cardiac activity monitoring system. Limnol. Oceanogr. Meth. 2014, 12, 670–679. [Google Scholar] [CrossRef]
  23. Kuklina, I.; Lozek, F.; Cisar, P.; Kouba, A.; Kozak, P. Crayfish can distinguish between natural and chemical stimuli as assessed by cardiac and locomotor reactions. Environ. Sci. Pollut. Res. 2018, 25, 8396–8403. [Google Scholar] [CrossRef]
  24. Kuramoto, T. Cardiac Activation and Inhibition Involved in Molting Behavior of a Spiny Lobster. Experientia 1993, 49, 682–685. [Google Scholar] [CrossRef]
  25. Yonkos, L.T.; Fisher, D.J.; Wright, D.A.; Kane, A.S. Pathology of fathead minnows (Pimephales promelas) exposed to chlorine dioxide and chlorite. Mar. Environ. Res. 2000, 50, 267–271. [Google Scholar] [CrossRef]
  26. Chupani, L.; Zuskova, E.; Stara, A.; Velisek, J.; Kouba, A. Histological changes and antioxidant enzyme activity in signal crayfish (Pacifastacus leniusculus) associated with sub-acute peracetic acid exposure. Fish. Shellfish Immun. 2016, 48, 190–195. [Google Scholar] [CrossRef]
  27. Chupani, L.; Stara, A.; Velisek, J.; Zuskova, E. Evaluation of the toxic effect of peracetic acid on grass carp (Ctenopharyngodon idella) juveniles. Neuroendocrinol. Lett. 2014, 35, 86–92. [Google Scholar]
  28. Straus, D.L.; Meinelt, T.; Farmer, B.D.; Beck, B.H. Acute toxicity and histopathology of channel catfish fry exposed to peracetic acid. Aquaculture 2012, 342, 134–138. [Google Scholar] [CrossRef]
Figure 1. Heart rate of crayfish P. leniusculus four and half hours before molting. The fluctuating line shows heart rate, beats per minute (bpm).
Figure 1. Heart rate of crayfish P. leniusculus four and half hours before molting. The fluctuating line shows heart rate, beats per minute (bpm).
Water 12 00063 g001
Figure 2. Chlorine dioxide (ClO2) concentration and crayfish mortalities during the monitoring period. The stars ( Water 12 00063 i001) indicate crayfish mortalities; the fluctuating line indicates levels of ClO2 concentration; the solid horizontal line indicates level of ClO2 concentration 0.2 mg L−1; the punctuated vertical line divides exposure period in two parts: First period (89 days), when high ClO2 concentrations (up to 0.2 mg L−1) were found five times and four crayfish died; and the second period (113 days), when high ClO2 concentrations occurred 23 times and 21 crayfish died.
Figure 2. Chlorine dioxide (ClO2) concentration and crayfish mortalities during the monitoring period. The stars ( Water 12 00063 i001) indicate crayfish mortalities; the fluctuating line indicates levels of ClO2 concentration; the solid horizontal line indicates level of ClO2 concentration 0.2 mg L−1; the punctuated vertical line divides exposure period in two parts: First period (89 days), when high ClO2 concentrations (up to 0.2 mg L−1) were found five times and four crayfish died; and the second period (113 days), when high ClO2 concentrations occurred 23 times and 21 crayfish died.
Water 12 00063 g002
Figure 3. Examples of heart rate of five crayfish P. leniusculus during the day and night period, 5th to 6th of June 2017 with ClO2 concentrations of 0.25 mg L−1 and 0.19 mg L−1, respectively. The fluctuating line indicates heart rate in beats per minute (bpm), while the two vertical lines distinguish night- and day-time.
Figure 3. Examples of heart rate of five crayfish P. leniusculus during the day and night period, 5th to 6th of June 2017 with ClO2 concentrations of 0.25 mg L−1 and 0.19 mg L−1, respectively. The fluctuating line indicates heart rate in beats per minute (bpm), while the two vertical lines distinguish night- and day-time.
Water 12 00063 g003
Figure 4. Examples of heart rate of five crayfish P. leniusculus two days before mortality occurred. The line indicates heart rate in beats per minute. Daily ClO2 (mg L−1) concentration are situated above the graphs, time of death is indicated by the vertical arrow.
Figure 4. Examples of heart rate of five crayfish P. leniusculus two days before mortality occurred. The line indicates heart rate in beats per minute. Daily ClO2 (mg L−1) concentration are situated above the graphs, time of death is indicated by the vertical arrow.
Water 12 00063 g004
Figure 5. Life duration after Cmax exposure of crayfish three experimental groups: Group one got Cmax on day 4 ± 2; Group two on day 13 ± 1; and Group three was exposed to Cmax on day 38 ± 6. Life duration was: 9 ± 7 days, 16 ± 8 days, and 5 ± 2 days, respectively. There was a significant difference (p < 0.05) in life duration between groups: Individuals from Group two in general lived more than twice as long as crayfish from Groups one and three after exposure to Cmax.
Figure 5. Life duration after Cmax exposure of crayfish three experimental groups: Group one got Cmax on day 4 ± 2; Group two on day 13 ± 1; and Group three was exposed to Cmax on day 38 ± 6. Life duration was: 9 ± 7 days, 16 ± 8 days, and 5 ± 2 days, respectively. There was a significant difference (p < 0.05) in life duration between groups: Individuals from Group two in general lived more than twice as long as crayfish from Groups one and three after exposure to Cmax.
Water 12 00063 g005
Table 1. Crayfish division based on the day of exposure to maximum concentration of ClO2 (Cmax); life duration after Cmax treatment over the entire exposure period; and N—Number of crayfish in each group. Data presented as means ± SD.
Table 1. Crayfish division based on the day of exposure to maximum concentration of ClO2 (Cmax); life duration after Cmax treatment over the entire exposure period; and N—Number of crayfish in each group. Data presented as means ± SD.
Crayfish GroupNCmax of ClO2, mg L−1Ordinal Day, When Cmax OccurredExposure Period Before Mortality, DaysLife Duration After Cmax Treatment, Days
1130.21 ± 0.044 ± 213 ± 89 ± 7
260.26 ± 0.0213 ± 129 ± 816 ± 8
360.29 ± 0.0138 ± 643 ± 75 ± 2
Table 2. Average heart rate expressed as beats per minute of all monitored crayfish from the three groups during the ClO2 exposure period. Mean ± SD.
Table 2. Average heart rate expressed as beats per minute of all monitored crayfish from the three groups during the ClO2 exposure period. Mean ± SD.
Crayfish GroupDay Heart Rate, bpmMaxMinNight Heart Rate, bpmMaxMinDay Versus Night Heart Rate, p-Value
153 ± 141092552 ± 14117200.54
250 ± 13892052 ± 1391260.37
347 ± 14922048 ± 1592230.68

Share and Cite

MDPI and ACS Style

Malinovska, V.; Ložek, F.; Kuklina, I.; Císař, P.; Kozák, P. Crayfish as Bioindicators for Monitoring ClO2: A Case Study from a Brewery Water Treatment Facility. Water 2020, 12, 63. https://doi.org/10.3390/w12010063

AMA Style

Malinovska V, Ložek F, Kuklina I, Císař P, Kozák P. Crayfish as Bioindicators for Monitoring ClO2: A Case Study from a Brewery Water Treatment Facility. Water. 2020; 12(1):63. https://doi.org/10.3390/w12010063

Chicago/Turabian Style

Malinovska, Viktoriia, Filip Ložek, Iryna Kuklina, Petr Císař, and Pavel Kozák. 2020. "Crayfish as Bioindicators for Monitoring ClO2: A Case Study from a Brewery Water Treatment Facility" Water 12, no. 1: 63. https://doi.org/10.3390/w12010063

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop