A non-invasive technique for sequential collection of earthworm (<i>Lumbricus terrestris</i>) leukocytes during subchronic immunotoxicity studies

Eyambe, George S.; Goven, Arthur J.; Fitzpatrick, Lloyd C.; Venables, Barney J.; Cooper, Edwin L.

doi:10.1258/002367791780808095

Cited by 217 publications

(90 citation statements)

References 11 publications

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“…Coelomic cells were obtained using a non-invasive extrusion method according to Eyambe et al (1991) with some modifications. Polychaetes were individually placed in 15 ml glass tubes containing 10 ml of 3% ethanol and 1,0 mg/ml EDTA in Hanks' balanced salt solution (H6648, Sigma Italy).…”

Section: Biological Analysesmentioning

confidence: 99%

Comparative Effects of Ingested PVC Micro Particles With and Without Adsorbed Benzo(a)pyrene vs. Spiked Sediments on the Cellular and Sub Cellular Processes of the Benthic Organism Hediste diversicolor

Gomiero

Strafella

Pellini³

et al. 2018

Front. Mar. Sci.

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Plastic micro litter represents an emerging contaminant as well as a multiple stress agent in aquatic environments. Microplastics are found even in the remote areas of the world. Together with their occurrence in all environmental compartments, there is a growing concern about their potential to adsorb pollutants co-occurring in the environment. At present, little is known about this source of exposure for aquatic organisms in the benthic environment. Exposure conditions were set up to mimick the contribution of microplastics through different exposure routes. Potential biological effects resulting from these exposures were investigated in the model organism Hediste diversicolor, an annelid worm. Cellular effects including alterations of immunological responses, lysosomal compartment changes, mitochondrial activity, oxyradical production and onset of genotoxicity were assessed in coelomocytes while temporary and permanent effects of oxidative stress were also performed at tissue level. In this study polyvinylchloride (PVC) microparticles were shown to adsorb benzo(a)pyrene with a time and dose-dependent relationship. The elevated bioavailability of the model pollutant after ingestion induced a clear pattern of biological responses. Toxicity mainly targeted impairment of cellular functioning and genotoxicity in H. diversicolor coelomocytes, while permanent effects of oxidative stress were observed at tissue level. Coelomocytes responded fast and with a higher degree of sensitivity to the adverse stimuli. The results showed that microplastic particles in sediments may play a significant role as vectors for organic pollutants. The highest adverse responses were observed in those H. diversicolor exposed to sediments spiked with PVC particles pre-incubated with B[a]P when compared against sediments spiked with B[a]P and plastic microparticles separately.

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Section: Biological Analysesmentioning

confidence: 99%

Comparative Effects of Ingested PVC Micro Particles With and Without Adsorbed Benzo(a)pyrene vs. Spiked Sediments on the Cellular and Sub Cellular Processes of the Benthic Organism Hediste diversicolor

Gomiero

Strafella

Pellini³

et al. 2018

Front. Mar. Sci.

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“…The coelomocytes of E. fetida after 15 and 30 days of exposure were obtained as described by Eyambe et al (1991). The worms were rinsed in saline solution (4°C) and gut contents were expelled by softly pressing the posterior part.…”

Section: Comet Assaymentioning

confidence: 99%

Biomarkers responses of the earthworm Eisenia fetida to acetochlor exposure in OECD soil

Xiao

Song

et al. 2006

Chemosphere

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“…Consequently, much effort has been undertaken towards the development of tests aimed at the evaluation of cellular responses at specific levels and in particular with L. terrestris (Rodriguez-Grau et al 1989, Eyambe et al 1991, Goven et al 1994, a widespread endogenous species by comparison with E. fetida. An important step in the development of new in vitro toxicity tests is the establishment of appropriate and standardized techniques, such as for coelomocyte isolation and maintenance.…”

Section: Discussionmentioning

confidence: 99%

“…A year later, Cooper (1974) proposed the recovery of 1. terrestris coelomocytes by puncture through the integument with a sharpened pasteur pipette. More recently, a non-invasive extrusion technique for the harvesting of 1. terrestris coelomocytes was proposed by Rodriguez-Grau et al (1989), a technique fully described by Eyambe et al (1991). Its novelty was in part the use of ethanol as an irritating agent and the addition of the mucolytic agent guaiacol glyceryl ether (GGE).…”

mentioning

confidence: 99%

“…In this study, in addition to modifications of the ethanol extrusion technique of Eyambe et al (1991), we propose the use of a low-vacuum holding device that eliminates the contamination of the cell preparation by faecal material released by the earthworms during the extrusion process. Furthermore, being successfully tested on 1. terrestris, E. fetida and O. tyrtaeum, the extrusion protocol proved to be highly adaptable to earthworms of different sizes and species.…”

mentioning

confidence: 99%

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Extrusion of earthworm coelomocytes: comparison of the cell populations recovered from the species Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum

Diogène¹,

Dufour²,

Poirier³

et al. 1997

Lab Anim

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SummaryCoelomocytes were extruded from three earthworm species: Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum. Featuring a simple low-vacuum holding device, the proposed methodology allows the recovery of cells with minimum risk of contamination by faecal material. The viability of O. tyrtaeum coelomocytes was highly reproducible (average 93%), with an average yield of 0.92 x 10 6 viable cells per earthworm. Cell viability for 1. terrestris and E. fetida averaged -68% but the cell yields were higher (respectively 1.67 x 10 6 and 1.28 x 10 6 ). Large inter-individual differences in cell yields were observed with 1. terrestris. Flow cytometric analyses indicated species to species differences in cell populations.Coelomocytes from E. fetida were the smallest with -57% of the total viable cells recovered being monitored between 2 and 10 tim. Large granulated cells I~20 tim) were detected in fairly large proportions in 1. terrestris and O. tyrtaeum [-52 and -96%, respectively I while they were less abundant in E. fetida (-9%). Using the vital dye neutral red to assess functional integrity, average cellular uptakes were significantly higher for 1. terrestris and O. tyrtaeum than for E. fetida (2.94, 2.66 and 0.64 tig/2 x 10 5 cells, respectively). In summary, the extrusion methodology herein described is applicable for the recovery of coelomocytes from a wide range of earthworm sizes and species. Moreover, this study strengthens the fact that extruded coelomocytes could be used for the evaluation of cell dysfunction and/or cell death following an in vitro and/or in vivo treatment. Keywords

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A non-invasive technique for sequential collection of earthworm (Lumbricus terrestris) leukocytes during subchronic immunotoxicity studies

Cited by 217 publications

References 11 publications

Comparative Effects of Ingested PVC Micro Particles With and Without Adsorbed Benzo(a)pyrene vs. Spiked Sediments on the Cellular and Sub Cellular Processes of the Benthic Organism Hediste diversicolor

Comparative Effects of Ingested PVC Micro Particles With and Without Adsorbed Benzo(a)pyrene vs. Spiked Sediments on the Cellular and Sub Cellular Processes of the Benthic Organism Hediste diversicolor

Biomarkers responses of the earthworm Eisenia fetida to acetochlor exposure in OECD soil

Extrusion of earthworm coelomocytes: comparison of the cell populations recovered from the species Lumbricus terrestris, Eisenia fetida and Octolasion tyrtaeum

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