soil and whole tissue [41]. Microbial diversity increased over time with the presence
of earthworms in soil as well as the overall enzymatic load in soil. Enzymatic
responses (AChE, GST and CEs) in valsartan-exposed L. terrestris were mostly
affected after 7 days and stabilised thereafter. Toxicity due to personal care products
(PCPs) was evaluated in E. andrei after a short term (24, 48 and 72 h) using the filter
paper contact test to several concentrations of triclocarban as modifications on CAT
and GST activities [42]. Exposure to triclosan (together with its metabolite methyltriclosan) was evaluated in E. andrei under longer (28 days) soil exposures as DNA
damage occurrence in coelomocytes [43]. To gain in environmental relevance, the
toxicity associated with drug chemical mixtures was evaluated in E. fetida exposed
ex situ to cosmetic sludge (CS) and foundry sands (FS) from industrial activities,
using DNA damage and coelomic cell composition as endpoints, after 1 h and in vivo
after 7- and 14-day exposure trials [44]. The suitability of using a non-invasive
method for coelomocyte extrusion and genotoxicity evaluation to variable percentage of water leachates, 1.5, 3, 6, 12.5, 25, 50 and 100%, was demonstrated as ex situ
but also in vivo in coelomocytes as soon as after 7-day FS and CS leachate
exposures. Toxicity evaluation of soil affected by livestock farming activities
(mostly made of trace metals, veterinary pharmaceuticals and pesticide residues)
was validated in the epi-endogeic earthworm Amynthas gracilis using sublethal
responses in AChE and SOD activities in whole body tissue and lysosomal integrity
(LMS) and DNA damage in coelomocytes after 1-, 7- and 14-day exposures [45].
7 Biomarkers of Nanomaterial Exposure
As anticipated, studies screening for the toxicity associated with this class of
emerging contaminants has been carried out mostly in the model earthworms
adopted in soil ecotoxicology testing (Eisenia spp.). Toxicity associated with
multiwalled carbon nanotubes (MWCNTs) was assessed in E. fetida exposed to
contaminated soil for up to 14 days, and, in addition to traditional endpoints
(survival and reproduction), a set of biomarkers including morphometric measures,
LMS, MT and AChE activity were considered [46]. A large proportion of soil
toxicity assessment studies with these nano-contaminants using earthworms refer
to engineered nanomaterial such as zinc oxide (ZnO). To select a few, a recent
comprehensive study with E. fetida considered an in vivo approach on the effects of
several concentrations of ZnO for up to 28 days in the responses on oxidative stressrelated biomarkers (ROS formation, LPO levels and SOD activity). This in vivo
approach was complemented by means of in vitro exposures to this nanomaterial in
the coelomic fluid as ROS formation and LDH activity measures [47]. Since
nanomaterial-induced toxicity is usually ROS-related, the highest ROS content
and LPO levels were found on day 28 at the highest dose nano-ZnO concentration
(250 mg/kg). Similarly, ZnO nanoparticles alone and combined with the pesticide
chlorpyrifos were assessed in E. andrei in a laboratory soil exposure for up to
28 days and included responses of oxidative stress biomarkers (CAT and GST),
Biomarkers in Earthworms
327
of earthworms in soil as well as the overall enzymatic load in soil. Enzymatic
responses (AChE, GST and CEs) in valsartan-exposed L. terrestris were mostly
affected after 7 days and stabilised thereafter. Toxicity due to personal care products
(PCPs) was evaluated in E. andrei after a short term (24, 48 and 72 h) using the filter
paper contact test to several concentrations of triclocarban as modifications on CAT
and GST activities [42]. Exposure to triclosan (together with its metabolite methyltriclosan) was evaluated in E. andrei under longer (28 days) soil exposures as DNA
damage occurrence in coelomocytes [43]. To gain in environmental relevance, the
toxicity associated with drug chemical mixtures was evaluated in E. fetida exposed
ex situ to cosmetic sludge (CS) and foundry sands (FS) from industrial activities,
using DNA damage and coelomic cell composition as endpoints, after 1 h and in vivo
after 7- and 14-day exposure trials [44]. The suitability of using a non-invasive
method for coelomocyte extrusion and genotoxicity evaluation to variable percentage of water leachates, 1.5, 3, 6, 12.5, 25, 50 and 100%, was demonstrated as ex situ
but also in vivo in coelomocytes as soon as after 7-day FS and CS leachate
exposures. Toxicity evaluation of soil affected by livestock farming activities
(mostly made of trace metals, veterinary pharmaceuticals and pesticide residues)
was validated in the epi-endogeic earthworm Amynthas gracilis using sublethal
responses in AChE and SOD activities in whole body tissue and lysosomal integrity
(LMS) and DNA damage in coelomocytes after 1-, 7- and 14-day exposures [45].
7 Biomarkers of Nanomaterial Exposure
As anticipated, studies screening for the toxicity associated with this class of
emerging contaminants has been carried out mostly in the model earthworms
adopted in soil ecotoxicology testing (Eisenia spp.). Toxicity associated with
multiwalled carbon nanotubes (MWCNTs) was assessed in E. fetida exposed to
contaminated soil for up to 14 days, and, in addition to traditional endpoints
(survival and reproduction), a set of biomarkers including morphometric measures,
LMS, MT and AChE activity were considered [46]. A large proportion of soil
toxicity assessment studies with these nano-contaminants using earthworms refer
to engineered nanomaterial such as zinc oxide (ZnO). To select a few, a recent
comprehensive study with E. fetida considered an in vivo approach on the effects of
several concentrations of ZnO for up to 28 days in the responses on oxidative stressrelated biomarkers (ROS formation, LPO levels and SOD activity). This in vivo
approach was complemented by means of in vitro exposures to this nanomaterial in
the coelomic fluid as ROS formation and LDH activity measures [47]. Since
nanomaterial-induced toxicity is usually ROS-related, the highest ROS content
and LPO levels were found on day 28 at the highest dose nano-ZnO concentration
(250 mg/kg). Similarly, ZnO nanoparticles alone and combined with the pesticide
chlorpyrifos were assessed in E. andrei in a laboratory soil exposure for up to
28 days and included responses of oxidative stress biomarkers (CAT and GST),
Biomarkers in Earthworms
327