Ecotoxicology

, Volume 21, Issue 4, pp 1102–1111 | Cite as

Effects of metal pyrithione antifoulants on freshwater macrophyte Lemna gibba G3 determined by image analysis

  • Hideo Okamura
  • Luvsantsend Togosmaa
  • Takuya Sawamoto
  • Keiichi Fukushi
  • Tomoaki Nishida
  • Toshio Beppu
Article

Abstract

Copper pyrithione (CuPT2) and zinc pyrithione (ZnPT2) are two popular antifouling agents that prevent biofouling. Research into the environmental effects of metal pyrithiones has mainly focused on aquatic animal species such as fish and crustaceans, and little attention has been paid to primary producers. There have been few reports on residues in environmental matrices because of the high photolabile characteristics of the agents. Residue analyses and ecological effects of the metabolites and metal pyrithiones are not yet fully understood. This study was undertaken to assess the effects of CuPT2, ZnPT2, and six metabolites (PT2: 2,2′-dithio-bispyridine N-oxide, PS2: 2,2′-dithio-bispyridine, PSA: pyridine-2-sulfonic acid, HPT: 2-mercaptopyridine N-oxide, HPS: 2-mercaptopyridine, and PO: pyridine N-oxide) on a freshwater macrophyte. A 7-day static bioassay using axenic duckweed Lemna gibba G3 was performed under laboratory conditions. Toxic effects of test compounds were assessed by biomass reduction and morphological changes were determined in image analysis. Concentrations of ZnPT2 and CuPT2 and those of PT2 and HPT in the medium were determined by derivatizing 2,2′-dithio-bispyridine mono-N-oxide with pyridine disulfide/ethylene diamine tetra-acetic acid reagent that was equimolar with pyrithione. The toxic intensity of the compounds was calculated from the measured concentrations after 7-day exposure. ZnPT2, CuPT2, PT2, and HPT inhibited the growth of L. gibba with EC50 ranging from 77 to 140 μg/l as calculated from the total frond number as the conventional index, whereas the other four metabolites had less effect even at 10 mg/l. The presence of the former four toxic derivatives resulted in abnormally shaped and unhealthily colored fronds, whose size was about 20% of the control fronds. EC50, calculated from the healthy frond area determined in image analysis, ranged from 10 to 53 μg/l. Thus, image analysis as part of a duckweed bioassay can detect the toxic effects of pyrithione derivatives with 3–10 times higher sensitivity than the traditional index.

Keywords

Abnormal Copper Duckweed Frond area Frond color Mercaptopyridine 

Abbreviations

AFS treaty

International Convention on the Control of Harmful Anti-Fouling Systems on Ships

CuPT2

Copper pyrithione, 2-mercaptopyridine N-oxide copper salt

DCM

Dichloromethane

DMSO

Dimethyl sulfoxide

EC50

50%-Effect concentration

EDTA

Ethylene diamine tetra-acetic acid

HFA

Healthy frond area

HFN

Healthy frond number

HPLC

High-performance liquid chromatography

HPS

2-Mercaptopyridine

HPT

2-Mercaptopyridine N-oxide

IQL

Instrumental quantification limit

IFA

Individual frond area, same as frond size

MeCN

Acetonitrile

NOEC

No-observed-effect concentration

PDS

Pyridine disulfide, same as PS2

PO

Pyridine N-oxide

PPMD

2,2′-Dithio-bispyridine mono-N-oxide

PS2

2,2′-Dithio-bispyridine

PSA

Pyridine-2-sulfonic acid

PT2

2,2′-Dithio-bispyridine N-oxide

SGR

Specific growth rate

TFA

Total frond area

TFN

Total frond number

ZnPT2

Zinc pyrithione, 2-mercaptopyridine N-oxide zinc salt

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Hideo Okamura
    • 1
  • Luvsantsend Togosmaa
    • 2
  • Takuya Sawamoto
    • 2
  • Keiichi Fukushi
    • 2
  • Tomoaki Nishida
    • 3
  • Toshio Beppu
    • 4
  1. 1.Research Center for Inland Seas, Kobe UniversityKobeJapan
  2. 2.Graduate School of Maritime Sciences, Kobe UniversityKobeJapan
  3. 3.Faculty of Agriculture, Shizuoka UniversityShizuokaJapan
  4. 4.Faculty of Life and Environmental Science, Teikyo University of ScienceYamanashiJapan

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