Daphnia magna responses to fish kairomone and chlorpromazine exposures

https://doi.org/10.1016/j.cbi.2020.109123Get rights and content

Highlights

  • Chlorpromazine (CPZ) and Fish kairomone (FK) caused functional alteration in D. magna.

  • Feeding rate was decreased in all treatment conditions.

  • Oxygen consumption was only affected in the mixture treatment (CPZ + FK).

  • For life history traits and phototactic behavior the effects of FK dominated those of CPZ.

Abstract

To avoid being preyed, organisms must be able to identify predatory threats by sensing molecules released by predators (kairomones), and to employ effective strategies to prevent detection by predators. Furthermore, in the wild, organisms are also exposed to chemicals that may alter their behavioral traits, such as neuroactive pharmaceuticals. Considering the co-occurrence of both types of chemicals, their possible interaction needs to be studied. To address this topic, the aim of this study was to verify the effects of fish kairomone (FK – a chemical associated to putative predation by fish) and chlorpromazine (CPZ - neuroactive pharmaceutical drug, environmental contaminant), isolated and in combination, in different functional endpoints of Daphnia magna, such as oxygen consumption, feeding rate, behavior and reproduction. Among these endpoints, oxygen consumption was only affected by the combination of compounds (FK + CPZ). On the other hand, feeding rate was affected by all treatments, being lower than control. For life history traits and phototactic behavior, the effects of FK predominated over the ones caused by CPZ exposure, incrementing the reproductive output of females, leading to greater population growth rates and increasing negative phototactic behaviour.

Introduction

Among aquatic animals, chemical signals can mediate many intra- and inter-specific interactions [1] and even visual species respond strongly to such cues [2]. In aquatic systems, chemical cues are recognized as being highly important for the detection of predators, thereby determining prey survival [3]. Alarm cues and predatory cues (i.e. kairomones) are the first two primary stimuli extracted from the sensory landscape as information about the potential predation [4]. In addition, many predator–prey relationships are mediated by the release and detection of kairomones. The importance of these agents as chemical mediators is extreme, since they are responsible for the induction of different responses, such as modifications in behavior, feeding, and reproduction [5].

A biological interaction during which one organism kills and feeds on another organism is one of predation This relationship not only shapes natural ecosystems but is also likely to occur in anthropogenically impacted environments [6]. To avoid becoming someone's meal, an organism must be able to identify predatory threats and employ effective strategies to avoid being detected by predators. In the event that such avoidance fails, animals can trigger behaviors to prevent predators from attacking, and also deploy tactics that will increase their chances of surviving the attack, in the case if it does occur [7]. This can be achieved by avoiding the same habitat that is occupied by predators, or by being active at different times of the day, or by altering the behavioral activity in response to changes in predatory risk [7], among others. Besides these behavioral alterations, predator-induced responses are important aspects of the population and even community ecology of freshwater zooplankton [8]. This happens since a good perception of danger and a correct anti-predator strategy should increase the chances of survival at the population level, regardless the species-specific reaction [9].

Furthermore, in the wild individuals interact with other individuals of the same (or of other species) and are exposed to chemicals. Among the array of environmental pollutants, one may find pharmaceutical compounds, which can alter population dynamics, animal behavior, reproduction, health and other features [10,11,96]. In terms of behavioral alterations, especially those drugs with neuroactive properties are particularly troublesome. Pharmaceutical drugs are a global concern in aquatics environments, since the number of these chemicals that are found in the wild is ever increasing [12]. One of these pharmaceuticals is chlorpromazine (CPZ), a low-cost aliphatic phenothiazine, widely available and used in treatments of psychiatric disorders worldwide, including schizophrenia [13]. Its primary antipsychotic activity is due to the blockade of dopamine (D2) receptors in the mesolimbic pathway of the brain [14]. This pharmaceutical has been shown to cause toxicological effects in non-target organisms, such as macroinvertebrates [15], macrophytes [16], and fish [17], and it has been found in different aquatic compartments in concentrations ranging from 1 to 364 ng/L [18,19].

The toxic effects of environmental chemicals needs a better and more realistic comprehension about their consequences and their impact; to attain this purpose, the organisms must be analyzed in more complex interaction scenarios, mimicking the actual and more realistic conditions of the ecosystem in which organisms live in [20]. Ecotoxicological studies focusing on the crosstalk between predation avoidance (and the effects of chemicals that act as chemical signals during this intercourse) and pharmaceuticals are adequate to study the joint contribution of natural and anthropogenic stressors. Despite the increasing number of studies dealing with the effects on living organisms caused by a large set of both natural and anthropogenic stressors, the characterization of these two combined factors is still scarce, despite being recognized as very important to understand how environmental contamination can affect the natural responses of aquatic animals to predation. Daphnia species are sensitive to a large number of substances and can be easily cultured under laboratory conditions, being therefore very useful in ecotoxicology as a model organism [21]. In addition, Daphnia species seem particularly sensitive to chemicals that are released by their predators, and somehow involved in their predation. The adaptive responses induced by predator kairomones are a key element in the modulation of individual fitness, which is the result of the contribution of both survival and reproduction success. Therefore, the success of a population is the consequence of individual survival and fitness, consequently, for the maintenance of Daphnia populations in aquatic ecosystems, the protection conferred by kairomone-mediated antipredator defenses is vital [22].

Daphnia species are ecologically important due to their role in many aquatic food webs. Daphnia act as primary consumer, and thereby control populations of primary producers; in addition, they also provide a food source for many secondary consumers [1]. Consequently, the responses of Daphnia towards aquatic contamination and/or kairomones may affect the entire ecosystem. To address this topic, the aim of this paper was to verify the effects of fish kairomone (indicative of predation) and chlorpromazine (environmental contaminant), alone and in combination, in different functional endpoints in Daphnia magna.

Section snippets

Chemicals

Chlorpromazine hydrochloride (CAS: 69-09-0) was purchased from Sigma-Aldrich (USA) with 98% of purity. All other used reagents had analytical purity.

Daphnia culture and maintenance

Clone P132,85 (used in all definitive experiments) was obtained from two generations of intraclonal mixes within clone P1, isolated from a small pond which contained fish (located at Driehoeksvijver, Heusden, The Netherlands; animals were isolated in August 1986). According to Ref. [23] organisms from this clone, in the presence of fish kairomones,

Results

Obtained results indicated that the tested compound was stable in ASTM water since concentrations in freshly prepared solutions (N = 4, Mean ± SE, 1092.8 ± 10.4 μg/L) were similar to those aged for 24 (1073.5 ± 5.6 μg/L) and 48 h (1063.2 ± 11.5 μg/L).

The oxygen consumption was lower (F3;12 = 4.091, p < 0.05) than control in the combination (FK + CPZ) (Fig. 1). On the other hand, feeding rates (Fig. 2) were significantly smaller (p < 0.05) in all treatments, during the exposure (F3;16 = 4.825)

Discussion

Zooplanktonic organisms, and in particular, Daphnia species, have been shown to exhibit responses to predator-specific chemicals (i.e. kairomones) in different parameters, as life history and behavioral traits [33]. However, the association of this natural stressor with anthropogenic ones (i.e. pharmaceuticals) is less studied, and this study brings some perspectives about this potential association. The here-adopted strategy is a comprehensive framework that includes a series of multiple

Conclusions

In summary, the tested natural and anthropogenic stressors (FK and CPZ, respectively) acted jointly decreasing oxygen consumption rates, swimming speed and aggregation, but operated independently on feeding. Concerning life history traits and phototactic behavior, the effects of FK predominated over those of CPZ, making females reproduce to a greater extent and hence having greater population growth rates and also decreasing positive phototactic behavior. Finally, according to the analyzed

CRediT authorship contribution statement

Gilberto Dias de Alkimin: Conceptualization, Methodology, Formal analysis, Investigation, Visualization, Writing - original draft, Writing - review & editing. Bruno Nunes: Visualization, Writing - review & editing, Supervision, Project administration, Funding acquisition. Amadeu MVM. Soares: Writing - review & editing, Supervision, Project administration, Funding acquisition. Marina Bellot: Validation, Investigation, Writing - review & editing. Cristian Gómez-Canela: Validation, Investigation,

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgements

This work was funded by the Spanish Ministry of Science and Innovation project (CTM2017-83242-R). Thanks are due to the Portuguese Science and Technology Foundation (FCT) supported the doctoral fellowship (Ref.a CRM:0047512) of Gilberto Dias de Alkimin. Thanks, are also due to ECO-R-pharmplast - Ecotoxicity of realistic combinations of pharmaceutical drugs and microplastics in marine ecosystems, Fundação para a Ciência e a Tecnologia, FCT (reference POCI-01-0145-FEDER-029203). Thanks are also

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