Energy, Ecology and Environment

, Volume 3, Issue 2, pp 81–86 | Cite as

Cyclodextrins as effective tools to reduce the toxicity of atrazine

  • Adneia de Fátima Abreu Venceslau
  • Fabio Eduardo dos Santos
  • Aline de Fátima Silva
  • Denise Alvarenga Rocha
  • Ademir José de Abreu
  • Carlos Jaime
  • Larissa Fonseca Andrade-Vieira
  • Luciana de Matos Alves Pinto
Original Article
  • 267 Downloads

Abstract

Atrazine (ATZ) is an agrochemical that is still widely used in the Americas to control intrusive weeds in large monocultures. However, its intrinsic toxicity can cause diseases of the endocrine and nervous systems. Cyclodextrins (CDs) are molecular carriers that can be employed to reduce the toxicity of ATZ. In this work, CDs (α, β, and γ) were anchored on silica, forming a hybrid material (CDSI). Lettuce (Lactuca sativa) was used as a model organism to evaluate the toxicity of the following treatments: ATZ; ATZ/α-CD; ATZ/β-CD; ATZ/γ-CD; ATZ/α-CDSI; ATZ/β-CDSI; and ATZ/γ-CDSI. The greatest chromosomal aberrations (CA) and nuclear abnormalities (NA) in the lettuce were observed with non-complexed ATZ. Reductions of CA ranged from 21% for ATZ/α-CD to 59% for ATZ/γ-CDSI, compared to non-complexed ATZ. In the case of NA, the decreases ranged from 29% for ATZ/β-CDSI to 68% for ATZ/α-CD, compared to non-complexed ATZ. The new synthesized CDSI material was found to be a viable option for reducing the toxicity of atrazine herbicide.

Graphical Abstract

Keywords

Herbicide Hybrid material Cyclodextrins Inclusion complex Cytotoxicity 

1 Introduction

Atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) is a selective triazine class herbicide used to control a broad spectrum of intrusive weeds. It is a weak base (pKa = 1.7) and its mechanism of action is related to the inhibition of photosynthetic activity in the chloroplast membrane, due to binding to the polyphenol oxidase of complex B of Photosystem II, disrupting the Hill reaction and preventing the flow of electrons (LeBaron et al. 2008).

ATZ is known to be toxic toward various living organisms. It affects the endocrine systems of mammals, binding to the growth hormone receptor (Fakhouri et al. 2010). It has also shown potential genotoxicity in the fish Carassius auratus (Cavas 2011). Studies in rats indicated an association between ATZ and decreases in testicular sperm production and cell viability (Abarikwu et al. 2012), in addition to causing myocardial angiogenesis (Rajkovic et al. 2014). In humans, the effects of ATZ have been found to include the induction of apoptosis in SH-SY5Y cells and the occurrence of neurodegenerative diseases (Abarikwu and Farombi 2015).

The inappropriate use of herbicides and insecticides is a common cause of food contamination (Markovic et al. 2010). ATZ is an emerging contaminant that was banned in Europe in 2004, being frequently found in soil, surface waters, and subterranean waters, but is still used in the Americas (European Commission 2004). Given its continuing use, studies have investigated techniques to reduce the toxicity of ATZ using modified release systems with ATZ encapsulated in carriers such as microspheres of poly(hydroxybutyrate-co-hydroxyvalerate) (Grillo et al. 2010) or nanocapsules of poly(ε-caprolactone) (Grillo et al. 2012).

Cyclodextrins (CDs) are among the carriers that have the potential for reducing the toxicity of herbicides. These substances are cyclic oligosaccharides composed of d-glucose molecules connected by α-1,4 glycosidic bonds and are obtained from the enzymatic degradation of starch by the cyclodextrin glycosyl transferase (CGTase) enzyme. Three types of CDs are commonly found in the environment: α, β, and γ-cyclodextrins, composed of 6, 7, and 8 glucose units, respectively (Crini 2014). The characteristics of CDs mean that they present excellent biocompatibility and the ability to encapsulate a wide range of molecules, as well as strong functionalization capacity (Zhang and Ma 2013).

Cyclodextrins can be immobilized on a variety of magnetic or polymeric supports, further enhancing their properties (Faraji 2005). One of the available supports is silica, which when combined with CDs has many potential applications, including the incorporation and modified release of agrochemicals such as ATZ (Carvalho and Pinto 2012). The hybrid materials formed by the combination of a CD and silica (CDSI) offer advantages over materials synthesized with organosilanes, including low cost and reduced toxicity (Baracho et al. 2015).

Evaluation of the effectiveness of ATZ complexes with CD, whether or not attached to silica, in reducing the toxicity of the herbicide, compared to free ATZ, can be assessed in trials employing higher plants, as described by Grant (1999). These assays enable assessment of the damage to an organism and its DNA caused by chemical agents (Andrade-Vieira et al. 2012). Tests of the effects of chemical substances on germination, initial seedling development, and the cell cycle have been widely used in environmental monitoring studies and for the assessment of contamination risks (Palmieri et al. 2014). These procedures are recognized and validated for this purpose by agencies such as the United States Environmental Protection Agency (US EPA) and the Organization for Economic Cooperation and Development (OECD) (Grant 1999; OECD 2003; US EPA 1996).

The advantages of using plant models in environmental toxicity studies are their low cost, easy handling, and sensitivity (Andrade et al. 2010), as well as their similarity to animal models, including human cells (Palmieri et al. 2016). Plant models do not need to be scrutinized by bioethics committees, in contrast to models involving animal sacrifice (Andrade-Vieira et al. 2014), in accordance with twenty-first-century norms in the area of toxicology, which emphasize the use of organisms other than animals in experimental procedures (Hartung 2009). Among the available plant models, Lactuca sativa L. can be used for macroscopic analyses (considering germination and early development) as well as microscopic evaluations of alterations in the cell cycle, which enable determination of the mechanism of action of the chemical agent (Aragão et al. 2015).

The purpose of the present work was to investigate the effectiveness of ATZ/CD and ATZ/CDSI inclusion complexes in reducing the toxicity of the herbicide atrazine in non-target organisms.

2 Materials and methods

2.1 Functionalization of silica with CDs

The CDs employed were obtained from Sigma (α-CD: C4642; β-CD: C4767; γ-CD: C4892). Silica gel (60 G, pore volume 0.65–0.85 mL g−1, specific surface area 350–450 m2 g−1), citric acid (99.5% purity), and xylol were obtained from Vetec. Anchoring of the CDs on the silica surface was performed following the methodology described by Carvalho et al. (2014). The product (CDSI) was filtered, dried, macerated, and stored in a freezer prior to subsequent use.

2.2 Preparation of the ATZ/CD and ATZ/CDSI inclusion complexes

The inclusion complexes were obtained following the methodology described by Carvalho and Pinto (2012). Briefly, equimolar amounts of ATZ and CD or CDSI were dissolved appropriately and kept under agitation. The solvent was then removed using a rotary evaporator and the solid products obtained were macerated, resuspended in ultrapure water, and lyophilized prior to subsequent use.

2.3 Bioassays using the L. sativa model

The protocol described by Aragão et al. (2015) was followed for the macroscopic analyses: germination index (GI), germination rate index (GRI), and root growth (RG); and the microscopic analyses: mitotic index (MI), chromosomal aberrations (CA), and nuclear abnormalities (NA). The test solutions used were ATZ/α-CD, ATZ/β-CD, ATZ/γ-CD, ATZ/α-CDSI, ATZ/β-CDSI, and ATZ/γ-CDSI, all at concentrations of 3 × 10−3 g L−1. The negative and positive controls were pure water and non-complexed ATZ, respectively.

Briefly, the seeds of L. sativa (variety Veronica) were disposed into a filter paper on Petri dishes (9 cm ø) with 3 mL of the treatment solutions. Each Petri dish contained 30 seeds, and there were 5 Petri dishes (replication) per treatment. The number of germinated seed (root protrusion) was counted each 8 until 48 h of exposure. The root length was obtained with a digital caliper after 120 h of exposure.

2.4 Statistical analysis

The parameters evaluated in the L. sativa assays (GI, GRI, RG, MI, CA, and NA) were submitted to analysis of variance (ANOVA), and the means were compared to the positive control (ATZ) using Tukey’s test (p < 0.05).

3 Results and discussion

The inclusion complexes produced using the CDs and the hybrid CDSI materials showed no significant interference for the parameters related to germination (GI and GRI). In the case of RG, after 120 h of exposure, all the treatments showed a significantly smaller root size, compared to the negative control (distilled water) (Table 1). Inhibition of root growth, relative to pure ATZ, was 23.10% for α-CD, 34.97% for β-CD, 38.75% for γ-CD, 18.17% for α-CDSI, 30.97% for β-CDSI, and 33.52% for γ-CDSI (Table 1).
Table 1

Analysis of the initial development of Lactuca sativa seedlings exposed to different atrazine treatments

Treatment

GI (%)a

GRIa

RGa

Atrazine

88.00 ± 5.05

12.35 ± 0.78

25.93 ± 0.97

ATZ/α-CD

88.00 ± 6.91

11.99 ± 0.98

19.94* ± 0.35

ATZ/β-CD

89.30 ± 6.83

12.23 ± 1.91

16.87* ± 0.83

ATZ/γ-CD

88.70 ± 4.47

12.58 ± 0.78

15.88* ± 0.33

ATZ/α-CDSI

88.00 ± 6.91

11.97 ± 0.96

21.22* ± 0.90

ATZ/β-CDSI

89.30 ± 7.23

12.24 ± 1.86

17.90* ± 1.64

ATZ/γ-CDSI

89.30 ± 5.47

12.69 ± 0.83

17.24* ± 1.34

Negative control

91.30 ± 7.67

13.00 ± 0.78

26.06 ± 0.61

GI germination index, GRI germination rate index, RG root growth, ATZ atrazine, CD cyclodextrin, CDSI cyclodextrin anchored to silica

Values followed by (*) indicate significant difference (Tukey’s test, p < 0.05) compared to the negative control (water)

aMeans followed by standard deviations

Among the macroscopic parameters (GI, GRI, and RG) evaluated using the L. sativa model, those related to germination (GI and GRI) are, according to Valerio et al. (2007), those that are least sensitive. The RG parameter is more effective in indicating the toxicity of the compound tested, since it is directly affected by delayed germination, which is reflected in a lower GRI value (Mauro et al. 2014). Hence, the significant decreases observed for ATZ complexed in CD and CDSI showed that the complexes were effective in enhancing the herbicidal effect of ATZ.

The genotoxicity of ATZ does not imply that it is herbicidal. Genotoxic effects are considered when the compound induces DNA damage. The herbicidal effect is considered where the compound inhibits the development of target plants. The plant development process involves both increases on the number of cells, which implies in enhancement on MI or elongation of cells, which depends on turgor pressing extensions and other proteins found on the cell wall. The herbicidal effects of ATZ were improved by the CD and CDSI as it decreases the RG. But in cellular level, it reduces ATZ genotoxicity as nuclear abnormities, and chromosome aberrations reduce subsequently. Thus, the RG is reduced for CD and CDSI as a consequence of problems on photosynthesis. This herbicidal effect was enhanced by CD and CDSI treatments.

Root growth is an important parameter in toxicological bioassays employing plant models, since it can be related to the parameters determined in the cytogenetic tests. The mitotic index (MI), which reflects the frequency of cell division, is a karyokinetic parameter directly related to the rate of root growth (Andrade et al. 2010).

Table 2 shows the results of the microscopic MI evaluation, with the percentages of each phase of mitotic division. Lower MI values were obtained for the cells treated with ATZ complexed using CD or CDSI, compared to treatment with pure ATZ. However, a significant reduction (of 17%), relative to free ATZ, was only observed for the ATZ/γ-CD treatment (Table 2). According to Fiskesjö (1993), this reduction is not biologically significant, because only mitodepressive compounds that reduce the frequency of cells in the division by more than 50% are considered cytotoxic.
Table 2

Analysis of the cellular cycle of meristematic Lactuca sativa cells exposed to different atrazine treatments

Treatment

Mitotic phase (%)

Mitotic indexa

Chromosomal aberrations (%)a

Nuclear abnormalities (%)a

Prophase

Metaphase

Anaphase

Telophase

Atrazine

56.70

19.90

11.13

12.27

7.32 ± 0.004

0.82 ± 0.0030*

11.45 ± 0.003b

ATZ/α-CD

53.31

22.40

12.10

12.19

6.68 ± 0.009

0.64 ± 0.0020*

3.65 ± 0.010*bc

ATZ/β-CD

52.51

20.56

13.00

13.92

6.40 ± 0.012

0.65 ± 0.0010*

6.46 ± 0.040*bc

ATZ/γ-CD

52.69

21.54

12.76

13.02

6.04 ± 0.005*

0.55 ± 0.0020*

5.14 ± 0.030*bc

ATZ/α-CDSI

57.43

19.79

10.75

12.03

7.21 ± 0.012

0.50 ± 0.0020*

5.15 ± 0.030*bc

ATZ/β-CDSI

55.83

20.05

9.81

14.31

7.30 ± 0.007

0.53 ± 0.0010*

8.15 ± 0.050*bc

ATZ/γ-CDSI

55.95

19.61

12.54

11.90

7.36 ± 0.008

0.34 ± 0.0020*

6.23 ± 0.020*bc

Negative control

56.29

18.14

11.17

14.40

7.85 ± 0.007

0.02 ± 0.0004

2.77 ± 0.020c

ATZ atrazine, CD cyclodextrin, CDSI cyclodextrin anchored to silica

Values followed by (*) indicate significant difference (Tukey’s test, p < 0.05) compared to the negative control (water)

aMeans followed by standard deviations

bSignificant in relation NC

cSignificant different in relation to ATZ

Despite not exhibiting cytotoxicity, according to the change in mitotic division rate, ATZ was shown to be genotoxic, because it increased the frequency of chromosomal aberrations leading to cell death.

The use of pure ATZ resulted in a 41-fold increase in the frequency of cells with abnormalities (Table 2). As pointed out by Fernandes et al. (2007), chromosomal aberrations are important parameters for determining the mechanisms of action of a particular chemical compound. Srivastava and Mishra (2009) analyzed the genotoxic effects of commercial ATZ in Allium cepa and Vicia faba and observed that ATZ induced chromosomal and nuclear abnormalities, with the most frequent being bridges, c-metaphase, and sticky chromosomes.

The earlier results were in agreement with the data obtained in the present work. Cells exposed to ATZ showed non-oriented chromosomes (Fig. 1a), chromosome fragments (Fig. 1b), sticky chromosomes (Fig. 1c), c-metaphase (Fig. 1d), and anaphase bridges (Fig. 1e). Among these abnormalities, sticky chromosomes were most frequent (Fig. 2). Chromosomal stickiness is one of the main abnormalities that, according to El-Ghamery et al. (2003), can initiate the processes leading to cell death. The frequency of condensed (pyknotic) nuclei, with stronger coloration than the normal nuclei (Fig. 1f), increased 4.13-fold for the treatment with pure ATZ, compared to water (Table 2). Pyknotic nuclei, such as those shown in Fig. 1f, are cytological markers of cell death (Andrade-Vieira et al. 2011), and in this case could have arisen due to the cytotoxicity of ATZ.
Fig. 1

Examples of different chromosomal abnormalities observed in meristematic root cells of Lactuca sativa exposed to atrazine inclusion complexes. a Non-oriented chromosome (arrow) in metaphase; b chromosome fragment (arrow) in anaphase; c sticky metaphase; d c-metaphase; e anaphase bridge; f condensed nuclei (arrows), showing stronger pigmentation than normal nuclei (head arrow). Bar 5 μm

Fig. 2

Chromosomal abnormalities observed for the treatments with free and complexed atrazine

When ATZ was complexed with CD or CDSI, there were decreases in the frequencies of chromosomal aberrations and condensed nuclei. For CA, the decreases were 21% for ATZ/α-CD, 21% for ATZ/β-CD, 33% for ATZ/γ-CD, 39% for ATZ/α-CDSI, 36% for ATZ/β-CDSI, and 59% for ATZ/γ-CDSI, compared to free ATZ. For NA, the percentage decreases were 68% for ATZ/α-CD, 44% for ATZ/β-CD, 55% for ATZ/γ-CD, 55% for ATZ/α-CDSI, 29% for ATZ/β-CDSI, and 46% for ATZ/γ-CDSI, compared to free ATZ. Among the complexes analyzed, ATZ/γ-CDSI showed the greatest reduction in the CA frequency (Table 2), while ATZ/α-CD showed the greatest reduction for condensed nuclei (Table 2). No differences were observed between ATZ complexed with the CDs and the CDSIs.

In tests with Allium cepa, Grillo et al. (2012) found lower rates of genotoxicity when triazine herbicides (ametryn, atrazine, and simazine) were encapsulated in poly (ε-caprolactone) nanocapsules. Pereira et al. (2014) found lower genotoxicity toward A. cepa cells when ATZ was incorporated in poly (ε-caprolactone) nanoparticles, in agreement with the present results and the findings of Grillo et al. (2010, 2012). Studies such as these are extremely important from the environmental point of view, because they offer an inexpensive way to mitigate problems of contamination caused by the widespread use of herbicides.

Grillo et al. (2010) also studying a controlled release system in which ATZ was encapsulated in poly (hydroxybutyrate-co-hydroxyvalerate) microspheres observed a significant reduction of genotoxicity in L. sativa cells after 24 h exposure of the roots to the herbicide. In this study, atrazine showed a dose-dependent inhibition of mitotic index (MI) and induced chromosomal aberrations (bridges, fragments, and chromosome delay). The results showed that free atrazine induces genotoxic effects on the roots of L. sativa and that the encapsulated ATZ, that is, the modified release of agrochemicals, can reduce the ATZ genotoxicity, corroborating with our results. ATZ is an herbicide with phytotoxic and genotoxic activities, whose toxicity can be reduced by the formation of inclusion complexes with cyclodextrins and cyclodextrins anchored to silica. Among the samples tested, the most promising was γ-CDSI, which provided the greatest decrease in atrazine genotoxicity.

Assays performed using L. sativa as a plant model, with evaluation of the initial development of the seedlings and alterations in the cell cycle, were shown to be effective in demonstrating the decrease of ATZ genotoxicity after complexation. Among the samples tested, the most promising was γ-CDSI, which provided the greatest decrease in atrazine genotoxicity.

Notes

Acknowledgements

The authors thank the Cytogenetics Laboratory of DBI/UFLA for providing access to the analytical equipment. Financial support was provided by FAPEMIG (#APQ-00687-13) and CAPES (Science without Frontiers Program, #A107/2013). A.F.A.V. received a grant from CAPES to undertake this work.

References

  1. Abarikwu SO, Farombi EO (2015) Atrazine induces apoptosis of SH-SY5Y human neuroblastoma cells via the regulation of Bax/Bcl-2 ratio and caspase-3-dependent pathway. Pestic Biochem Physiol 118:90–98. doi: 10.1016/j.pestbp.2014.12.006 CrossRefGoogle Scholar
  2. Abarikwu SO, Pant AB, Farombi EO (2012) The protective effects of quercetin on the cytotoxicity of atrazine on rat Sertoli-germ cell co-culture. Int J Androl 35:590–600. doi: 10.1111/j.1365-2605.2011.01239.x CrossRefGoogle Scholar
  3. Andrade LF, Davide LC, Gedraite LS (2010) The effect of cyanide compounds, fluorides, aluminum, and inorganic oxides present in spent pot-liner on germination and root tip cells of Lactuca sativa. Ecotoxicol Environ Saf 73:626–631. doi: 10.1016/j.ecoenv.2009.12.012 CrossRefGoogle Scholar
  4. Andrade-Vieira LF, Gedraite LS, Campos JMS, Davide LC (2011) Spent Pot Liner (SPL) induced DNA damage and nuclear alterations in root tip cells of Allium cepa as a consequence of programmed cell death. Ecotoxicol Environ Saf 74:882–888. doi: 10.1016/j.ecoenv.2010.12.010 CrossRefGoogle Scholar
  5. Andrade-Vieira LF, de Campos JMS, Davide LC (2012) Effects of Spent Pot Liner on mitotic activity and nuclear DNA content in meristematic cells of Allium cepa. J Environ Manag 107:140–146. doi: 10.1016/j.jenvman.2012.04.008 CrossRefGoogle Scholar
  6. Andrade-Vieira LF, Botelho CM, Palmieri MJ, Laviola BG, Praça-Fontes MM (2014) Effects of Jatropha curcas oil in Lactuca sativa root tip bioassays. An Acad Bras Ciên 86:373–382CrossRefGoogle Scholar
  7. Aragão FB, Andrade-Vieira LF, Ferreira A, Costa AV, Queiroz VT, Pinheiro PF (2015) Phytotoxic and cytotoxic effects of Eucalyptus essential oil on Lactuca sativa L. Allelopathy J 35:259–272Google Scholar
  8. Baracho RV, Carvalho LB, Andrade JM, Venceslau AFA, Rocha DA, Pinto LMA (2015) Obtenção e caracterização de material híbrido entre sílica e ciclodextrinas. Quím Nova 38:1063–1067. doi: 10.5935/0100-4042.20150099 Google Scholar
  9. Carvalho LB, Pinto LMA (2012) Formation of inclusion complexes and controlled release of atrazine using free or silica-anchored β-cyclodextrin. J Incl Phenom Macrocycl Chem 74:375–381. doi: 10.1007/s10847-012-0125-9 CrossRefGoogle Scholar
  10. Carvalho LB, Carvalho TG, Magriotis ZM, Ramalho TC, Pinto LMA (2014) Cyclodextrin/silica hybrid adsorbent for removal of methylene blue in aqueous media. J Incl Phenom Macrocycl Chem 78:77–87. doi: 10.1007/s10847-012-0272-z CrossRefGoogle Scholar
  11. Cavas T (2011) In vivo genotoxicity evaluation of atrazine and atrazine-based herbicide on fish Carassius auratus using the micronucleus test and the comet assay. Food Chem Toxicol 49:1431–1435. doi: 10.1016/j.fct.2011.03.038 CrossRefGoogle Scholar
  12. Crini G (2014) Review: a history of cyclodextrins. Chem Rev 114:10940–10975. doi: 10.1021/cr500081p CrossRefGoogle Scholar
  13. El-Ghamery AA, El-Kholy MA, El-Yousser MAA (2003) Evaluation of cytological effects of Zn2+ in relation to germination and root growth of Nigella sativa L. and Triticum aestivum L. Mutat Res 537:29–41. doi: 10.1016/S1383-718(03)00052-4 CrossRefGoogle Scholar
  14. European Commission (2004) Commission decision of 10 March 2004 concerning the non-inclusion of atrazine in Annex I to Council Directive 91/414/EEC and the withdrawal of authorisations for plant protection products containing this active substance, 2004/248/EC. OJEU L78:53–55Google Scholar
  15. Fakhouri WD, Nuñez JL, Trail F (2010) Atrazine binds to the growth hormone-releasing hormone receptor and affects growth hormone gene expression. Environ Health Perspect 118:1400–1405. doi: 10.1289/ehp.0900738 CrossRefGoogle Scholar
  16. Faraji H (2005) β-Cyclodextrin-bonded silica particles as the solid-phase extraction medium for the determination of phenol compounds in water samples followed by gas chromatography with flame ionization and mass spectrometry detection. J Chromatogr 1087:283–288. doi: 10.1016/j.chroma.2005.06.009 CrossRefGoogle Scholar
  17. Fernandes TCC, Mazzeo DEC, Marin-Morales MA (2007) Mechanism of micronuclei formation in polyploidizated cells of Allium cepa exposed to trifluralin herbicide. Pestic Biochem Physiol 88:252–259. doi: 10.1016/j.pestbp.2006.12.003 CrossRefGoogle Scholar
  18. Fiskesjö G (1993) The Allium test—a potential standard for the assessment of environmental toxicity. Am Soc Test Mater 2:331–345Google Scholar
  19. Grant WF (1999) Higher plant assays for the detection of chromosomal aberrations and gene mutations a brief historical background on their use for screening and monitoring environmental chemicals. Mutat Res 624:107–112CrossRefGoogle Scholar
  20. Grillo R, Melo NFS, Lima R, Lourenço RW, Rosa AH, Fraceto LF (2010) Characterization of atrazine-loaded biodegradable poly(hydroxybutyrate-co-hydroxyvalerate) microspheres. J Polym Environ 18:26–32. doi: 10.1007/s10924-009-0153-8 CrossRefGoogle Scholar
  21. Grillo R, Santos NZP, Maruyamac CR, Rosa AH, Lima R, Fraceto LF (2012) Poly (ε-caprolactone) nanocapsules as carrier systems for herbicides: physico-chemical characterization and genotoxicity evaluation. J Hazard Mater 231:1–9. doi: 10.1016/j.jhazmat.2012.06.019 CrossRefGoogle Scholar
  22. Hartung T (2009) Toxicology for the twenty-first century. Nature 460(9):208–212. doi: 10.1038/460208a CrossRefGoogle Scholar
  23. LeBaron HM, McFarland JE, Burnside OC (2008) The triazine herbicides: a milestone in the development of weed control technology. In: LeBaron HM, McFarland JE, Burnside OC (eds) The triazine herbicides: 50 years revolutionizing agriculture. Elsevier, Amsterdam, pp 1–12Google Scholar
  24. Markovic M, Cupac S, Durovic R, Milinovic J, Kljajic P (2010) Assessment of heavy metal and pesticide levels in soil and plant products from agricultural area of Belgrade, Serbia. Arch Environ Contam Toxicol 58:341–351. doi: 10.1007/s00244-009-9359-y CrossRefGoogle Scholar
  25. Mauro MO, Pesarini JR, Marin-Morales MA, Monreal MTFD, Monreal ACD, Mantovani MS, Oliveira RJ (2014) Evaluation of the antimutagenic activity and mode of action of the fructooligosaccharide inulin in the meristematic cells of Allium cepa culture. Genet Mol Res 13(3):4808–4819CrossRefGoogle Scholar
  26. Organization for Economic Cooperation and Development – OECD (2003) Terrestrial plant test: 208: seedling emergence and seedling growth test. Guideline for the testing of chemicals proposal for updating guideline 208Google Scholar
  27. Palmieri MJ, Luber J, Andrade-Vieira LF, Davide LC (2014) Cytotoxic and phytotoxic effects of the main chemical components of spent pot-liner: a comparative approach. Mutat Res Genet Toxicol Environ Mutagen 763:30–35. doi: 10.1016/j.mrgentox.2013.12.008 CrossRefGoogle Scholar
  28. Palmieri MJ, Andrade-Vieira LF, Campos JMS, Gedraite LS, Davide LC (2016) Cytotoxicity of Spent Pot Liner on Allium cepa root tip cells: a comparative analysis in meristematic cell type on toxicity bioassays. Ecotoxicol Environ Saf 133:442–447. doi: 10.1016/j.ecoenv.2016.07.016 CrossRefGoogle Scholar
  29. Pereira AE, Grillo R, Mello NF, Rosa AH, Fraceto LF (2014) Application of poly (epsilon-caprolactone) nanoparticles containing atrazine herbicide as an alternative technique to control weeds and reduce damage to the environment. J Hazar Mater 268:207–215. doi: 10.1016/j.jhazmat.2014.01.025 CrossRefGoogle Scholar
  30. Rajkovic V, Kovac R, Koledin I, Matavulj M (2014) Atrazine-induced changes in the myocardial structure of peripubertal rats. Toxicol Ind Health 30–3:250–258. doi: 10.1177/0748233712456058 CrossRefGoogle Scholar
  31. Srivastava K, Mishra KK (2009) Cytogenetic effects of commercially formulated atrazine on the somatic cells of Allium cepa and Vicia faba. Pestic Biochem Physiol 93:8–12. doi: 10.1016/j.pestbp.2008.08.001 CrossRefGoogle Scholar
  32. United States Environmental Protection Agency – US EPA (1996) How to effectively recover free product at leaking underground storage tank sites—a guide for state regulators. EPA, WashingtonGoogle Scholar
  33. Valerio ME, Garcia JF, Peinado FM (2007) Determination of phytotoxicity of soluble elements in soils, based on a bioassay with lettuce (Lactuca sativa L.). Sci Total Environ 378:63–66. doi: 10.1016/j.scitotenv.2007.01.007 CrossRefGoogle Scholar
  34. Zhang J, Ma PX (2013) Cyclodextrin-based supramolecular systems for drug delivery: recent progress and future perspective. Adv Drug Deliv Rev 65(9):1215–1233. doi: 10.1016/j.addr.2013.05.001 CrossRefGoogle Scholar

Copyright information

© Joint Center on Global Change and Earth System Science of the University of Maryland and Beijing Normal University and Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Adneia de Fátima Abreu Venceslau
    • 1
  • Fabio Eduardo dos Santos
    • 2
  • Aline de Fátima Silva
    • 1
  • Denise Alvarenga Rocha
    • 1
  • Ademir José de Abreu
    • 3
  • Carlos Jaime
    • 1
    • 4
  • Larissa Fonseca Andrade-Vieira
    • 2
  • Luciana de Matos Alves Pinto
    • 1
  1. 1.Department of ChemistryFederal University of Lavras (UFLA)LavrasBrazil
  2. 2.Department of BiologyFederal University of Lavras (UFLA)LavrasBrazil
  3. 3.Department of AdministrationFaculty of Sciences and Technology of Campos Gerais (FACICA)Campos GeraisBrazil
  4. 4.Department of ChemistryUniversitat Autònoma de Barcelona (UAB)BarcelonaSpain

Personalised recommendations