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Reconfirmation of antimicrobial activity in the coelomic fluid of the earthwormEisenia fetida andrei by colorimetric assay

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Abstract

A novel tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium (MTS) was used in the assessment of antimicrobial activity in earthworm in the presence of phenazine methosulphate (PMS) as an electron coupling reagent. This activity was purified from the coelomic fluid of the earthworm (ECF),Eisenia fetida andrei (Oligochaeta, Lumbricidae, annelids) using a series of column chromatography techniques and was tested against three Gram-negative strains ofEscherichia coli, Pseudomonas aeruginosa, Aeromonas hydrophila and three Gram-positive strains ofStaphylococcus aureus, Bacillus megaterium, Arthrobacter sp., respectively. Only the pigment-free eluate of coelomic fluid of the earthworm (ECFPE) showed activity againstB. megaterium amongst three isolated active fractions. The anion (DEAE-52) exchange effluent of the ECFPE was reported to have the strongest activity againstP. aeruginosa amongst the three active fractions. The 20% acetonitrile eluate (AE) by Sep-Pak C18 cartridge was also tested and showed fair resistance againstE. coli, P. aeruginosa andArthrobacter sp., respectively.

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Abbreviations

AE:

Acetonitrile eluate

DPBS:

Dulbecco’s phosphate buffered saline

ECF:

coelomic fluid of the earth-worms

ECFPE:

ECF pigment-free eluate

MICs:

minimum inhibitory concentrations

MTS:

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrazolium

PMS:

phenazine methosulphate

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Author notes

  1. Weidong Pan

    Present address: Institute of Zoology, Beisihuan West Road 25, PO Box 27, 100080, Beijing, People’s Republic of China

Authors and Affiliations

  1. State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, People’s Republic of China

    Weidong Pan, Xianghui Liu, Feng Ge & Tao Zheng

  2. Bioelectromagnetic Laboratory, Institute of Electrical Engineering, Chinese Academy of Sciences, Beijing, People’s Republic of China

    Tao Zheng

Authors
  1. Weidong Pan
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  2. Xianghui Liu
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  3. Feng Ge
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  4. Tao Zheng
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Correspondence to Feng Ge.

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Pan, W., Liu, X., Ge, F. et al. Reconfirmation of antimicrobial activity in the coelomic fluid of the earthwormEisenia fetida andrei by colorimetric assay. J Biosci 28, 723–731 (2003). https://doi.org/10.1007/BF02708433

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  • DOI: https://doi.org/10.1007/BF02708433

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