Assessment of chromosomal damage induced by a deltamethrin-based insecticide in fish (Anguilla anguilla L.) – A follow-up study upon exposure and post-exposure periods

https://doi.org/10.1016/j.pestbp.2014.06.003Get rights and content

Highlights

  • The deltamethrin-based insecticide Decis® induced chromosomal damage.

  • The transience of the cytogenetic damage, measured as ENA induction, was demonstrated.

  • It was suggested a rapid metabolization/elimination of Decis® constituents by the fish.

  • An increased erythrocyte turnover was associated to the return of ENA to control levels.

  • Present findings highlighted the genetic hazard to fish associated to Decis® exposure.

Abstract

The pyrethroid insecticide Decis®, containing deltamethrin as active ingredient, is among the most popular broad-spectrum biocides, with wide application in agriculture and home pest control. The occurrence of deltamethrin in the aquatic environment is well-established, but the possible genotoxic effects of Decis® in non-target organisms, namely fish, remain unknown. Hence, this work aimed to evaluate the cytogenetic damaging potential of Decis® in European eel (Anguilla anguilla L.), adopting the erythrocytic nuclear abnormalities (ENAs) assay. In addition, it was intended to investigate the damage progression in the post-exposure period. The frequency of immature erythrocytes (IE) was also determined to provide indirect information on the erythrocyte catabolism and erythropoiesis rate. Fish were exposed to 17.5 and 35 μg L−1 of Decis® (equivalent to 0.05 and 0.1 μg L−1 of deltamethrin, respectively) during 1 and 3 days. Thereafter, fish were transferred to clean water and kept for 1, 7 and 14 days. The results demonstrated a clear potential to induce chromosomal damage following 3 days exposure, depicted in an ENA frequency increase for both Decis® concentrations. The transient nature of this cytogenetic damage was also demonstrated, as ENA frequency returned to the control level 1 and 7 days after cessation of the exposure, respectively for the higher and the lower Decis® concentration. Moreover, this response pattern suggested a rapid metabolization and elimination of the formulation constituents by A. anguilla, combined with an increased erythrocyte turnover in fish exposed to the higher Decis® concentration, as pointed out by the IE frequency rise. Overall, the demonstrated genotoxic properties of Decis® pointed out increased risk factors to fish exposed to this insecticide.

Introduction

The use of biocides to control harmful organisms is regarded as an unavoidable tool in several sectors, namely agriculture, forestry, industry and public health. Since they are intended to kill living organisms, numerous biocidal products have also an inherent capacity to cause adverse effects on humans or the environment. In this direction, it is of particular concern the contamination of aquatic systems, as these compounds are easily washed from surfaces, mainly due to natural processes as water runoff and soil leaching, reaching superficial waters also by spray-drift during applications. Therefore, it is undeniable that aquatic organisms, namely fish, will be exposed by dermal contact, breathing and water ingestion, ending particularly affected [1], [2].

Among the large variety of biocides, defined according to the organism against which are used, insecticides have triggered a considerable environmental apprehension due to their high toxicity, extensively demonstrated in non-target organisms [3], including fish [4], [5], [6]. Pyrethroids are among the most used insecticides worldwide and one of the most toxic group to fish and aquatic invertebrates [2], which has been explained by their lipophilicity and high rate of absorption through gills [7], [8]. Nonetheless, it has been also stated that the low doses commonly applied and short persistence in the environment can avoid the major adverse effects for those organisms [9]. Deltamethrin, the active ingredient of the commercial formulation Decis®, is a synthetic pyrethroid with insecticidal properties used to protect several fruit and vegetable crops, effective against a multiplicity of pests, which acts in the peripheral and central nervous systems of insects (interfering with the sodium channels) [9]. Deltamethrin has very low water solubility, strong adsorption to soil and sediments, and rapid degradation in the environment [9]. However, the deltamethrin-based commercial products, like Decis®, behave differently. Decis® is miscible with water due to the adjuvant (petroleum naphtha), which potentially increases the bioavailability of the active principle in the dissolved phase. The presence of deltamethrin in the aquatic environment was detected in water streams nearby rice cultivations on the island of Leyte (Philippines) in concentrations between 0.001 and 0.043 μg L−1 [10]. Additionally, a previous study reported 0.1 μg L−1 of deltamethrin in water samples collected near a potato field in Manitoba (Canada) [11].

Several studies have been conducted evaluating the sublethal toxicity of deltamethrin and related commercial formulations to fish. Thus, Decis® showed to induce alterations in biochemical and hematological parameters in Poecilia reticulata (e.g. glutathione S-transferase, lactate dehydrogenase and acetylcholinesterase activities) [5], Oncorhynchus mykiss and Cyprinus carpio (e.g. plasma glucose, erythrocyte count, hemoglobin content, hematocrit and plasma total proteins) [12], [13]. This formulation also caused histopathological effects in gills of Aphanius dispar [14], in gills and kidney of C. carpio [15], and in liver of Gambusia affinis [16].

It is well established that water contaminants, such as pesticides, can attack DNA and thus, the analysis of genetic damage in aquatic organisms is considered a suitable method for evaluating their environmental hazard [17]. Nevertheless, studies on the genotoxicity of Decis® or its active principle in fish remain scarce. Ansari et al. [18] assessed the genotoxic effect of deltamethrin in Channa punctata, reporting the induction of erythrocytic micronuclei and other nuclear abnormalities. Additionally, Grisolia [19] pointed out a higher vulnerability of fish to Decis®, comparing to mouse, since only fish (Tilapia rendalli) showed increased micronuclei frequency following exposure to this commercial insecticide.

Taking into account the most common applications of insecticides, their input to the aquatic systems is typically intermittent. Hence, and owing also to fish mobility, the exposure to this type of contaminants can be short and eventually followed by a period permanence in non-contaminated areas. Bearing this in mind, the assessment of genotoxic endpoints after removal of the contamination source appears as a crucial approach to determine the actual magnitude of risk posed by these biocides to fish. However, this perspective is absent in the majority of fish studies evaluating the genotoxicity of pesticides.

Considering the above statements and the gaps identified, it is relevant to further assess the genotoxic potential of Decis® to fish, clarifying the damage progression in the post-exposure period. Thus, the main goal of this work was to evaluate the genetic damage in the European eel (Anguilla anguilla L.), subjected to a short-term exposure (1 and 3 days) to 17.5 and 35 μg L−1 of Decis®, corresponding to 0.05 and 0.1 μg L−1 of deltamethrin, which should be considered environmentally realistic, regarding the levels of this pyrethroid detected in surface waters (see Refs. [10] and [11]). In addition, it was assessed the eventual genetic damage recovery after fish transfer to clean water (1–14 days post-exposure). The cytogenetic damage was evaluated through the erythrocytic nuclear abnormalities (ENAs) assay performed in peripheral erythrocytes. This methodology relies on the fact that genotoxins can promote chromosomic cleavage (clastogenicity) or even the complete loss of a chromosome, as well as dysfunction at the mitotic spindle level (aneugenicity) [20], [21], [22]. Assuming that those nuclear anomalies reflect irreparable lesions, the eventual recovery of the damage would be possible mainly by processes of cell turnover (replacement of defective cells with newly generated ones), purging the blood of abnormal erythrocytes. Therefore, the frequency of immature erythrocytes (IE) was also determined to survey the hematological dynamics, obtaining indirect information on the erythrocyte catabolism (hemocatheresis) and erythropoiesis rate.

Section snippets

Chemicals

The commercial formulation tested – Decis® – is distributed by Bayer CropScience Portugal, containing deltamethrin as the active ingredient at 25 g L−1 (2.85%), calcium 4-(4,6,8-trimethylnonan-3-yl)benzenesulfonate (1–5%), 2-methyl-1-propanol (1–5%), and naphtha (petroleum) as adjuvant/solvent (>25%). All the other chemicals were obtained from the Sigma–Aldrich Chemical Company (Madrid, Spain).

Test animals and experimental design

Eels with an average weight of 0.25 ± 0.02 g (glass eel stage) were captured at Minho river mouth, Caminha,

ENA assay

Concerning the first day of exposure, no significant alterations were found in ENA frequency, for both concentration groups of Decis® (Fig. 3), as well as in the different lesion categories analyzed individually (Table 1). Nevertheless, after 3 days of exposure, both concentrations caused significant ENA increase in comparison with the control (Fig. 3). The analysis of each individual nuclear lesion category demonstrated that K and L frequencies were significantly higher in D1 group (17.5 μg L−1

Discussion

The contamination of surface waters with genotoxic chemicals is likely to pose a serious threat to the health and survival of fish, since it can be on the basis of adverse effects such as changes in gene expression, cellular impairment and cancer. However, genotoxicity induction by pesticides in fish is a topic not entirely explored. On the other hand, since the use of deltamethrin and other pyrethroids is expected to increase worldwide, the ecotoxicologic assessment of their usage is

Conclusions

Overall, the present findings highlighted the genetic hazard to fish associated to the deltamethrin-based insecticide Decis®, since a potential to induce chromosomal damage following a short-term exposure (3 days) was clearly demonstrated.

The transient nature of the cytogenetic damage, measured as ENA induction, was also demonstrated. Thus, ENA frequency returned to the control level 1 and 7 days after cessation of the exposure, respectively for the higher and the lower Decis® concentration. The

Ethical statement

This study was conducted in accordance with the EU Directive 2010/63/EU on the protection of animals used for scientific purposes, under the supervision of a team member (Mário Pacheco) authorized by the competent authorities.

Conflict of interest

The authors declare that there are no conflicts of interest.

Acknowledgments

This work was supported by Fundação Ciência e Tecnologia (FCT; Government of Portugal) through the Research project PTDC/AAC-AMB/114123/2009 [co-financed by FCT/MCTES in its national budget component (PIDDAC) through the Investigation Fellowship (BI/CESAM/PTDC/AAC-AMB/114123/2009) and by the European Regional Development Fund (ERDF) through COMPETE – Thematic Factors of Competitiveness Operational Program (POFC)], as well as by Centre for Environmental and Marine Studies (CESAM).

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