A comparison of biomarkers in assessing the combined effects of pesticide mixtures on non-target soil invertebrates

Date
2004-12
Journal Title
Journal ISSN
Volume Title
Publisher
Stellenbosch : University of Stellenbosch
Abstract
ENGLISH ABSTRACT: Agricultural environments are usually contaminated with mixtures of antropogenically introduced chemicals as a result of pesticide spraying, which can affect beneficial, nontarget soil invertebrates, such as earthworms negatively. Most studies on mixture toxicity have focused on interactions of chemicals with similar structures and mechanisms. However, chemical mixtures may occur as conglomerates of diverse structures and toxicological mechanisms in the environment. This study was aimed at assessing the effects of pesticides singly, and in a mixture, on earthworms, using lifecycle parameters (growth and reproduction) and biomarkers (neutral red retention (NRR) assay and acetylcholinesterase (AChE) inhibition) as endpoints. Thus, to determine whether any interactions occurred between the pesticides as shown by the measured endpoints. Another aim was to validate the use of the chosen biomarkers for assessing mixture toxicity. The pesticides used were from three groups: organophosphates, heavy metal-containing pesticides and pyrethroids. From these three groups, four of the most commonly used pesticides in the orchards and vineyards of the Western Cape, South Africa, were chosen, namely chlorpyrifos (organophosphate), azinphos-methyl (organophosphate), copper oxychloride (heavy metal-containing fungicide) and cypermethrin (pyrethroid). Earthworms were exposed in the laboratory to a range of concentrations of chlorpyrifos and copper oxychloride singly, and in 1:1 mixtures of these pesticides in artificial soil, for four weeks. After the exposure period, the biomass change was determined as measure of growth, and cocoon production, hatching success and number of hatchlings per cocoon were determined as measures of reproduction. Growth (biomass change) and reproduction (cocoon production) were affected by the highest concentration treatment (20mg/kg) of chlorpyrifos, but copper oxychloride and the mixture of the two pesticides showed no observable effects on lifecycle parameters. Dose related effects on NRR times were however determined for both pesticides and the mixture. Dose related effects on AChE activity were found for chlopyrifos and the mixture of the two pesticides, but not for copper oxychloride. Short-term exposures (48 hours) of earthworms to the following pesticides in artificial groundwater: chlorpyrifos, copper oxychloride, azinphos-methyl, cypermethrin, chlorpyrifos-copper oxychloride, chlorpyrifos -azinphos-methyl and chlorpyrifos-cypermethrin, were done followed by the determination of AChE inhibition. Dose related effects were exhibited on the AChE activity of earthworms exposed to chlorpyrifos, a mixture of chlorpyrifos and copper oxychloride, azinphos-methyl, and a mixture of azinphos-methyl and chlorpyrifos. Copper oxychloride, cypermethrin and the mixture of chlorpyrifos and cypermethrin had no effect on AChE activity. Earthworms died at the highest exposure concentration of the mixture of chlopyrifos and cypermethrin. Results have shown that although the pesticides did not cause observable effects on lifecycle parameters, there were effects at subcellular and biochemical level, as shown by the biomarkers. Mixtures of pesticides, in some instances, affected earthworms differently from their single components, indicating interactions between the pesticides in mixtures, as shown by the measured endpoints. The NRR assay proved to be a good general biomarker of soil contamination, and the AChE activity could also be a valuable tool in assessing the effects of organophosphate mixtures and mixtures of organophosphates and pesticides from other groups.
AFRIKAANSE OPSOMMING: Nie-teiken organismes, soos erdwurms, word negatief beïnvloed deur mengsels van antropogeniese chemikalieë in landbou-omgewings. Die meeste studies wat handel oor die toksisiteit van chemiese mengsels het tot dusver gefokus op chemikalieë van dieselfde aard en met dieselfde meganismes van werking. Mengsels van chemiese stowwe kan egter as konglomerate van 'n verskeidenheid strukturele eienskappe en met verskillende toksiese meganismes in die omgewing aangetref word. Tydens die studie is gepoog om die effekte van enkel pestisiede sowel as mengsels daarvan op erdwurms te bestudeer, deur van lewensloop kenmerke (groei en voortplanting) en biomerkers (neutraalrooi retensietyd - NNR en inhibisie van asetielcholienesterase -AChE) as eindpunte gebruik te maak. 'n Verdere doel van die studie was om vas te stel of daar enige wisselwerkings tussen die verskillende pestisiede plaasvind, soos aangetoon deur die gemete eindpunte, en verder ook om die gebruik van die gekose biomerkers as maatstawwe van mengseltoksisiteit te evalueer. Die pestisiede wat gebruik is, is van drie verskillende groepe afkomstig: organofosfate, swaarmetale en piretroiede. Van hierdie drie groepe is vier van die pestisiede wat vry algemeen in boorde en wingerde in die Weskaap, Suid-Afrika, gebruik word, geïdentifiseer. Hierdie stowwe is chlorpyrifos (organofosfaat), azinphos-metiel (organofosfaat), koperoksichloried (swaarmetaalbevattende fungisied) en sipermetrien (piretroied). Erdwurms is in die laboratorium aan 'n reeks konsentrasies van chlorpyrifos en koperoksichloried as enkel toksikante en as 1:1 mengsels in kunsmatige grond, vir vier weke blootgestel. Voor en na die blootstellingsperiode is die biomassa van die wurms, as maatstaf van groei, bepaal en kokonproduksie, uitbroeisukses en getal nakomelinge per kokon bepaal as maatstawwe van voortplantingsvaardigheid. Groei (biomassaverandering) en voortplanting (kokonproduksie) is beinvloed deur behandeling met die hoogste konsentrasie (20 mg/kg) chlorpyrifos, terwyl geen effek van koperoksichloried of die mengsel van hierdie twee pestisiede gevind is nie. Daar is gevind dat beide die pestisiede, enkel en in die mengsel, die NRR tye beinvloed het. Die AChE aktiwiteit is beinvloed deur chlorpyrifos en die mengsel, maar nie deur die koperoksichloried nie. Korttermyn blootstellings van erdwurms (48 uur), in kunsmatige grondwater, van erdwurms aan chlorpyrifos, koperoksichloried, azinphos-metiel en sipermetrien as enkel toksikante en mengsels van chlorpyrifos-koperoksichloried, chlorpyrifos-azinphos-metiel en chlorpyrifos-sipermetrien, is gedoen en gevolg deur die bepaling van AChE inhibisie. Koperoksichloried, cypermetrien en die chlorpyrifos-sipermetrien mengsel het geen waarneembare effek op die AChE aktiwiteit gehad nie ?????. Die erdwurms wat blootgestel is aan die hoogste konsentrasie in die mengsel van chlorpyrifos-sipermetrien het doodgegaan. Die resultate het getoon dat die pestisiede nie in die korttermyn die lewensloopkenmerke in enige waarneembare mate geaffekteer het nie maar daar was effekte op sellulêre en biochemiese vlakke soos aangetoon deur die biomerkers. Sommige mengsels van die pestisiede het die erdwurms verskillend van die enkelstowwe geaffekteer. Daar het dus wisselwerking tussen sommige van die pestisiede wat in mengsels aangewend is, plaasgevind, soos aangetoon deur die gemete eindpunte. Die NRR toets, as breë-spektrum biomerker was 'n goeie maatstaf van kontaminasie in grond en daar is aanduidings dat die AChE aktiwiteit, as 'n spesifieke biomerker, 'n nuttige maatstaf kan wees om die effekte van organofosfaatmengsels en mengsels van hierdie chemiese groep en die van ander chemikalieë aan te toon.
Description
Thesis (MSc)--University of Stellenbosch, 2004.
Keywords
Soil invertebrates -- Effect of pesticides on, Earthworms -- Effect of pesticides on, Soil invertebrates -- Effect of chemicals on, Pesticides -- Environmental aspects, Theses -- Zoology, Dissertations -- Zoology
Citation