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Effects of 5-chloro-2-methyl-4-isothiazolin-3-one and other candidate biodiesel biocides on rat alveolar macrophages and NR8383 cells

  • Organ Toxicity and Mechanisms
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Abstract

Biocides are added to biodiesels to inhibit and remove microbial growth. The effects of 5-chloro-2-methyl-4-isothiazolin-3-one (CMIT), a candidate biodiesel biocide, were studied using freshly isolated rat alveolar macrophages (AM) and NR8383 cell line. CMIT markedly inhibited phagocytic oxidative burst as measured by zymosan-induced chemiluminescence, and cellular cytokine secretion as measured by zymosan-induced TNF-α secretion. The 50% inhibition concentration (LC50) for CMIT was 0.002–0.004 mM for both cellular functions. AM exposed to CMIT for as little as 2 min showed markedly inhibited functions that persisted for at least 5 h. Sodium metabisulfite was able to partially neutralize the inhibitory activity of CMIT. Cysteine and glutathione, when present at a molar ratio of 2–1 or higher against CMIT, were effective neutralizers, while serine, histidine, alanine, and albumin were without effect. When the AM testing system was used to compare the toxicity of CMIT against three other candidate biodiesel biocides, methylene dithiocyanate (MDC) was found to be of comparable toxicity to CMIT, 2-methyl-4-isothiazolin-3-one (MIT) was much less toxic, and dimethyl acetylenedicarboxylate (DMAD) was non-toxic. Because AM is among the first cell-type exposed to inhaled biodiesel aerosols, the result suggested that CMIT present in biodiesel may produce respiratory effects, and further investigations including animal studies are warranted.

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Notes

  1. Luminol is not readily soluble in incubation mixtures. The following procedure ensures complete solubilization. Luminol (17.7 mg) in 10 ml DMEM at 37 oC was sonicated to a fine suspension using a probe sonicator (Ultrasonic Processor, Cole-Parmer Instrument C., IL., USA). One volume of the suspension was added slowly to 9 volume of DMEM solution at 37 oC with vigorous mixing.

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Acknowledgments

We wish to thank Ka Lei Ku Karen Leingartner and Brita Nadeau for excellent technical assistance. This work was supported by the Canadian Clean Air Research Agenda (CARA) and by the Canadian Program of Energy Research and Development (PERD).

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Authors and Affiliations

  1. Environmental Heath Science and Research Bureau, Health Canada, 50 Columbine Driveway, Ottawa, K1A 0K9, Canada

    R. Poon & M. Rigden

  2. Water, Air and Climate Change Bureau, Health Canada, 260 Laurier Avenue W, Ottawa, K1A 0K9, Canada

    N. Edmonds, N. Charman & S. Lamy

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  1. R. Poon
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  2. M. Rigden
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  3. N. Edmonds
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  4. N. Charman
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Correspondence to R. Poon.

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Poon, R., Rigden, M., Edmonds, N. et al. Effects of 5-chloro-2-methyl-4-isothiazolin-3-one and other candidate biodiesel biocides on rat alveolar macrophages and NR8383 cells. Arch Toxicol 85, 1419–1427 (2011). https://doi.org/10.1007/s00204-011-0689-y

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