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Poster highlight: Environmental concentrations of the cocaine metabolite benzoylecgonine induced sublethal toxicity in the development of plants but not in a zebrafish embryo-larval model

These results were presented in the 25th European Meeting of SETAC  (6th May 2015) in a poster (WE293) and a brief speech (poster highlight, 15:55, room 112). / Estos resultados fueron presentados en la Europea de la SETAC en Barcelona (6 de Mayo) mediante un póster y una breve exposición oral (poster highlight 15:55, sala 112).

Download the presentation for the Poster spotlight WE293 – M Catala

See the extended abstract in a previous post.

See the poster by clicking in Poster BE SETAC 2015.

Download the poster by clicking in Poster BE SETAC 2015.

Your comments will be appreciated.

Extended abstract: Environmental concentrations of the cocaine metabolite benzoylecgonine induced sublethal toxicity in the development of plants but not in a zebrafish embryo-larval model

Catalá, M.1, García-Cambero, J.P.2, García-Cortés,H.1, Valcárcel Rivera, Y3.

1Department of Biology ande Geology, Physics and Inorganic Chemistry. Higher School of Science and Technology. Rey Juan Carlos University. Campus de Móstoles c/Tulipán s/n E-28933 Móstoles

2National Centre for Environmental Health. Institute of Health Carlos III. Ctra Majadahonda-Pozuelo km 2, 28220 Majadahonda, Madrid, Spain

3Yolanda Valcárcel Rivera. Grupo de Investigación en Salud Ambiental y Ecotoxicología. Area de Medicina Preventiva y Salud Pública. Facultad de Ciencias de la Salud. Universidad Rey Juan Carlos.
E-mail contact: myriam.catala@urjc.es

1.    Introduction

Several studies have found cocaine and its main active metabolite benzoylecgonine (BE) in the aquatic environment and natural waters [1] derived from their high consumption by humans and the inability of water treatment processes to eliminate it [2]. As an emerging substance, research on BE has primarily focused on improving methodologies for its determination, while hardly any study has tackled the impact that such waste can have on wild biota. Only a couple of works have addressed BE ecotoxicology on animals [3-4] and no data have been published of the ecotoxicity of BE on plants or microalgae.

Concerning fish, there is growing interest in the use of fish embryos as an alternative tool to examine the presence and potency of aquatic toxicants. On one hand, it is generally considered that initial phases of development are more susceptible to environmental pollution than adults, what is determinant for the viability of natural populations [5]. On the other hand, new endpoints, beyond simple lethality, have emerged to assist toxicity identification in fish embryos, including the study of developmental effects and sublethal effects relative to cardiotoxicity, hepatotoxicity or neurotoxicity among others.

A microbioassay based on the development of fern spores/gametophytes has been developed some years ago, turning into a robust and sensitive method for the assessment of the toxicity of industrial pollutants [6], pharmaceuticals [7] or environmental samples [8].

So, the aim of this publication is to provide information on the toxicity of environmental concentrations of BE during animal and vascular plant development. To evaluate fish toxicity, we have used a combined model (embryo-larvae) to identify sensitive biomarkers, e.g. those related to neurodevelopment [9]. For plant toxicity, we have used the fern spores model. Data from this study will contribute to a better understanding of the impact of such emerging substances in different organisms of the freshwater ecosystems, necessary for environmental risk assessment.

2.    Materials and methods

A stock solution of 1 mg mL-1 of BE in methanol was prepared. The concentrations (ranged from 10-4 to 10 µg L-1) were selected to cover the environmental concentrations of BE detected in fluvial systems. In order to assess the toxicity of BE on zebrafish embryos, several toxicological endpoints were monitored: viability, alterations in development (morphology, biometry and hatching), cardiotoxicity (presence of edema, impaired circulation and heartbeat frequency), and finally larvae locomotion. Two different endpoints were used to evaluate acute phytotoxicity (48 hours): mitochondrial activity for cell energetic status and DNA quantification for lethality.To evaluate subchronic phytotoxicity (168 hours), chlorophyll autofluorescence was also measured. The mitochondrial activity was measured according to with minor modifications [7]. For DNA quantification, samples were sent to the Genomic Service of Pompeu Fabra University for DNA quantification with the PicoGreen® method (QuantiFluor ST Fluorometer and Paradigm Detection Platform, Beckman). Chlorophyll autofluorescence was measured (λexc: 485nm, λem: 635 nm) in a SPECTRAFluor Plus microplate reader (Tecan Group Ltd. Männedorf, Switzerland).

3.    Results and discussion

3.1.     Toxicity during fish embryo development

In spite of the potential cardiotoxic and neurotoxic effects of cocaine, the main metabolite BE (0.01 to 1000 µg L-1) did not seem to affect the heartbeat rate -indicative of cardiotoxicity- nor the locomotion at day 6pf (behavior) in this zebrafish embryo-larval bioassay. Embryos exposed to BE did not show malformations, and their growth and hatching were comparable to the control group. The low toxicity of BE to zebrafish embryos is in line with the poor toxic activity of BE found in other experimental vertebrates, such as rats [10].

3.2.     Phytotoxicity during development

BE resulted to interfere with the development of gametophytes at so low concentrations as 0.001 µg L-1 affecting both metabolic and genetic processes. BE environmental concentrations have a significant effect on total DNA levels as well as a strong inhibitory effect on mitochondrial activity. The reduction of DNA levels (ca. 20%) could be attributed either to the death of part of the spore population or to a cytostatic effect of BE, causing a delay in spore cell cycle. Alkaloids in general and benzoic acid derivatives in particular, are were studied long ago for their allelopathic effects since all seeds and fruits known for their high alkaloid content are strong germination inhibitors [11]. Evenari classified cocaine as a strong germination inhibitor together with caffeine and codeine among others in dilutions of 1/200. BE has demonstrated the ability to disrupt cell function and probably the cell cycle that may, in the long term, lead to the abortion of spore germination or to abnormalities in the developed gametophyte.

4.  Conclusions

BE could pose an environmental risk for riparian ferns gametophyte recruitment and vascular plant germination and growth. These results agree with the allelophathic role described for alkaloids such as benzoylecgonine and their unspecific interference with plant germination. We conclude that the anthropogenic dispersion of alkaloid allelochemicals may pose a risk for biodiversity and irrigated food production that should be further investigated.

 References

  1. Mendoza a, López de Alda M, González-Alonso S, Mastroianni N, Barceló D, Valcárcel Y. 2014. Occurrence of drugs of abuse and benzodiazepines in river waters from the Madrid Region (Central Spain). Chemosphere. 95:247–55.
  2. Huerta-Fontela M, Galceran MT, Ventura F. 2008. Stimulatory drugs of abuse in surface waters and their removal in a conventional drinking water treatment plant. Environ. Sci. Technol. 42:6809–6816.
  3. Parolini M, Binelli A. 2013. Adverse effects induced by ecgonine methyl ester to the zebra mussel: a comparison with the benzoylecgonine. Environ. Pollut. 182:371–8.
  4. Pedriali A, Riva C, Parolini M, Cristoni S, Sheehan D, Binelli A. 2013. A redox proteomic investigation of oxidative stress caused by benzoylecgonine in the freshwater bivalve Dreissena polymorpha. Drug Test. Anal. 5:646–56.
  5. Bergek S, Ma Q, Vetemaa M, Franzén F, Appelberg M. 2012. From individuals to populations: Impacts of environmental pollution on natural eelpout populations. Ecotoxicol. Environ. Saf. 79:1–12.
  6. Marugán J, Bru D, Pablos C, Catalá M. 2012. Comparative evaluation of acute toxicity by Vibrio fischeri and fern spore based bioassays in the follow-up of toxic chemicals degradation by photocatalysis. J. Hazard. Mater. 213-214:117–122.
  7. Feito R, Valcárcel Y, Catalá M. 2013. Preliminary data suggest that venlafaxine environmental concentrations could be toxic to plants. Chemosphere. 90:2065–2069.
  8. Esteban S, Fernandez Rodriguez J, Diaz Lopez G, Nunez M, Valcarcel Y, Catala M. 2013. New microbioassays based on biomarkers are more sensitive to fluvial water micropollution than standard testing methods. Ecotoxicol. Environ. Saf. 93:52–59.
  9. García-Cambero JP, Catalá M, Valcárcel Y. 2012. River waters induced neurotoxicity in an embryo-larval zebrafish model. Ecotoxicol. Environ. Saf. 84:84–91.
  10. Morishima HO, Okutomi T, Ishizaki A, Zhang Y, Cooper TB. 2001. The disposition of benzoylecgonine in maternal and fetal rats. Neurotoxicol. Teratol. 23:247–253.
  11. Evenari M. 1949. Germination inhibitors. Bot. Rev. 15:153–194.

Acknowledgement – The authors thank the program “Projects on Health Research 2011-2012” FIS (PI11/00180)” of Carlos III Health Institute

Comments

These results were presented in the 25th European Meeting of SETAC next week (6th May) in a poster (WE293) and a brief speech (poster highlight, 15:55, room 112). / Estos resultados fueron presentados en la reunión Europea de la SETAC en Barcelona (6 de Mayo 2015) mediante un póster y una breve exposición oral (poster highlight 15:55, sala 112).

Download the poster highlight brief presentation

See the communication by clicking in Poster BE SETAC 2015

Download by clicking in Poster BE SETAC 2015

Your comments will be appreciated.

Environmental concentrations of the cocaine metabolite benzoylecgonine induced sublethal toxicity in the development of plants but not in a zebrafish embryo-larval model

These results will be presented in the 25th European Meeting of SETAC next week (6th May) in a poster (WE293) and a brief speech (poster highlight, 15:55, room 112). / Estos resultados serán presentados en la próxima reunión Europea de la SETAC en Barcelona (6 de Mayo) mediante un póster y una breve exposición oral (poster highlight 15:55, sala 112).

See the communication by clicking in Poster BE SETAC 2015

Download by clicking in Poster BE SETAC 2015

Your comments will be appreciated.

Elimination of drugs of abuse and their toxicity from natural waters by photo-Fenton treatment

These results will be presented in the 25th European Meeting of SETAC next week (5th May) in a poster (TU379) . / Estos resultados serán presentados en la próxima reunión Europea de la SETAC en Barcelona (5 de Mayo) .

See the communication by clicking in Poster Photo-Fenton SETAC 2015

Download by clicking in Poster Photo-Fenton SETAC 2015

We will appreciate your comments

Presentación de ToxAmb y exposición sobre cocaína en aguas fluviales

En el pasado FESCET II, congreso organizado por la Unión de Alumnos del Campus de Móstoles, Helena García, miembro de ToxAmb, dio una conferencia denominada “Cocaína en aguas fluviales”.

Comenzó con la presentación del grupo de investigación y explicó algunas definiciones básicas que permitieran seguir fácilmente el resto de la exposición a los alumnos presentes de tan diversas titulaciones.

Tras un breve repaso de los estudios sobre contaminantes emergentes (o microcontaminantes) llevados a cabo por ToxAmb, se centró en el caso del principal metabolito de la cocaína (benzoilecgonina) desde la perspectiva de la ecotoxicología.

Finalmente, resaltó la importancia de este tipo de estudios ya que la presencia de determinadas sustancias tales como disruptores endocrinos, drogas de abuso y fármacos y sus metabolitos en las aguas, pueden suponer un riesgo para la salud humana y la biodiversidad.

Homeopatía: #NoSinEvidencia

Es muy triste que la pseudociencia se abra paso por la legislación por el simple hecho del beneficio económico de algunos, que puede perjudicar la salud de otros muchos. La formulación de la filosofía de la Homeopatía en sí es claramente anticientífica: la dilución “infinita” del principio activo. Si alguno de los preparados vendidos bajo la firma de “Medicamento Homeopático”, por moda o creencia, funciona, nunca lo sabremos ¡se va a admitir legalmente que se vendan sin probar su eficacia!

Y hay otro problema mayor. La toxicología tradicionalmente ha admitido que una sustancia tiene un umbral de acción, es decir, una concentración mínima a la que ejerce su acción típica. Por ejemplo, se considera que un medicamento o incluso un veneno, no ejercerá efectos por debajo de una concentración, cantidad o dosis determinada. Sin embargo, recientes estudios, en los que yo misma y ToxAmb estamos implicados, demuestran que muchas sustancias pueden ejercer efectos biológicos por debajo de esas concentraciones determinadas en los ensayos de toxicidad/actividad clásicos. Y lo que es más importante, esos efectos pueden ser distintos e incluso opuestos a los conocidos y demostrados previamente. Este fenómeno se está haciendo especialmente patente cuando investigamos los efectos biológicos de los microcontaminantes. La explicación biológica tiene total lógica y fundamento en la fisiología celular, todas las células van a intentar defenderse de sustancias extrañas (xenobióticos) en cuanto las detecten. Y los modelos que definen estas respuestas biológicas se denominan “Modelos de Hormesis”. Pues bien, concentraciones consideradas inocuas para ciertas sustancias, pueden sorprendentemente ejercer ciertos efectos biológicos a concentraciones muchísimo más bajas. A veces beneficiosos, o a veces perjudiciales ¿quién lo sabe?

Las autoridades se han quedado muy tranquilas aprobando la seguridad de los preparados homeopáticos con la idea subyacente de que son inocuos, y por ende seguros. Todo lo más, asumen, constituyen un fraude para el consumidor y lo esperable es un “efecto placebo”. Pero se sospecha que parte de la reputación de estos preparados homeopáticos, cuya dilución no es infinita aunque sí alta (superior a partes por billón), provenga de la activación de sistemas de defensa celulares que responden a mechanismos horméticos (ver la abundante bibliografía científica reciente de A. Calabrese).

Concluyendo, podríamos encontrarnos con preparados homeopáticos que pueden afectar negativamente a la salud de algunas personas. Dichos efectos negativos se sumarían a la mera ineficacia, pudiendo agravar el estado de salud de un enfermo por falta de tratamiento. Por lo tanto reclamemos que todos los medicamentos sean testados para su eficacia y seguridad.

Si quieres unirte a esta campaña, puedes visitar el Blog Nosinevidencia, y compartir tu opinión.