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http://dx.doi.org/10.7847/jfp.2012.26.1.031

Comparison of microscopic counting and alamar blue assay to evaluate anti-protozoal effects against Azumiobodo hoyamushi that causes soft tunic syndrome to Halocynthia roretzi  

Lee, Jae-Geun (Department of Aquatic Life Medicine, Kunsan National University)
Zeon, Seung-Ryul (Department of Aquatic Life Medicine, Kunsan National University)
Park, Kyung-Il (Department of Aquatic Life Medicine, Kunsan National University)
Choi, Sang-Hoon (Department of Aquatic Life Medicine, Kunsan National University)
Park, Kwan Ha (Department of Aquatic Life Medicine, Kunsan National University)
Publication Information
Journal of fish pathology / v.26, no.1, 2013 , pp. 31-38 More about this Journal
Abstract
The edible ascidian, Halocynthia roretzi is a commercially important fisheries resource in Korea. However, there have been outbreaks of mass mortality due to soft tunic syndrome. It was discovered recently that the cause of death is infection by a protozoan parasite Azumiobodo hoyamushi. Alamar blue assay and microscopic counting were used to estimate anti-protozoal effects of 20 drugs having different action mechanisms. Through comparison of alamar blue assay and microscopic counting, 6 drugs were found to be potential in protozoan-killing effects: amphotericin B, formalin, hydrogen peroxide, bithionol, benzalkonium chloride, bronopol (24hr-$EC_{50}{\leq}20{\mu}g/ml$). The preliminary data can be used as a basis to develop anti-protozoal agents against A. hoyamushi.
Keywords
Halocynthia roretzi; Azumiobodo hoyamushi; microscopy; alamar blue and anti-protozoal drugs;
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Times Cited By KSCI : 2  (Citation Analysis)
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1 Ostland V. E., Byrne P. J., Speare D. J., Thorburn M. A., Cook A., Morrison D. and Ferguson H.W.: Comparison of formalin and chloramine-T for control of a mixed gill infection (bacterial gill disease and ichthyobodiasis) in rainbow trout. J. Aquat. Anim. Health., 7:118-123, 1995.   DOI
2 Park K. H., Zeon S. R., Lee J. G., Choi S. H., Shin Y. K. and Park K. I.: In vitro and in vivo efficacy of drugs against the protozoan parasite Azumiobodo hoyamushi that causes soft tunic syndrome in the edible ascidian Halocynthia roretzi. J. Fish Dis., in press, 2013
3 Raz B., Iten M., Bühler Y. G., Kaminsky R. and Brun R.: The alamar $blue^{(R)}$ assay to determine drug sensitivity of african trypanosomes (T.b. rhodesiense and T.b. gambiense) in vitro. Acta Trop., 68:139-147, 1997.   DOI   ScienceOn
4 Russo R., Curtis E. W. and Yanong R. P. E.: Preliminary investigations of hydrogen peroxide treatment of selected ornamental fishes and efficacy against external bacteria and parasites in green swordtails.
5 Saha A. K., Mukherjee T. and Bhaduri A.: Mechanism of action of amphotericin B on Leishmania donovani Promastigotes. Mol. Biochem. Parasitol., 19:195-200, 1986.   DOI   ScienceOn
6 Shinn A. P., Picón-Camacho S. M., Bron J. E., Conway D., Yoon G. H., Guo F. C. and Taylor N. G. H.: The anti-protozoal activity of bronopol on the key life-stages of Ichthyophthirius multifilliis Fouquet, 1876 (Ciliophora). Vet. Parasitol., 186:229-236, 2012.   DOI   ScienceOn
7 신윤경, 김현중, 박경일, 최민순, 전제천, 김응오: 통영산 물렁증 멍게 피막의 편모충 감염. 한국어병학회지, 24:197-204, 2011.   과학기술학회마을   DOI   ScienceOn
8 홍정표, 김영섭, 허성범: 온도자극 및 수용밀도에 따른 우렁쉥이 폐사. 한국양식학회지, 13:285-293, 2000.   과학기술학회마을
9 Florent R. L., Becker J. A. and Powell M. D.: Efficacy of bithionol as an oral treatment for amoebic gill disease in Atlantic salmon Solmo Salar(L.). Aquaculture., 270:15-22, 2007.   DOI   ScienceOn
10 Florent R. L., Becker J. A. and Powell M. D.: In vitro toxicity of bithionol and bithionol sulphoxide to Neoparamoeba spp., the causative agent of amoebic gill disease (AGD). Dis. Aquat. Org., 91:259-262, 2010.
11 Franzblau S. G., Witzig R. S., Mclaughlin J. C., Torres P., Madico G., Hernandez A., Degnan M. T., Cook M. B., Quenzer V. K., Ferguson R. M. and Gilman R. H.: Rapid, low-technology MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate alamar blue assay. J. Clin. Microbiol., 36:362-366, 1998.
12 Hirose H., Nozawa A., Kumagai A. and Kitamura S.: Azumiobodo hoyamushi gen. nov. et sp. nov. (Euglenozoa, Kinetoplastea, Neobodonida): A pathogenic kinetoplastid causing the soft tunic syndrome in ascidian aquaculture. Dis. Aquat. Organ., 97:227-236, 2012.   DOI   ScienceOn
13 Hamid R., Rotshteyn Y., Rabadi L., Parikh R. and Bullock P.: Comparison of alamar blue and MTT assays for high through-put screening. Toxicol. In Vitro., 18:703-710, 2004.   DOI   ScienceOn
14 Hirazawa N., Goto T. and Shirasu K.: Killing effect of various treatments on the monogenean Heterobothrium okamotoi eggs and oncomiracidia and the ciliate Cryptocaryon irritans cysts and theronts. Aquaculture., 223:1-13, 2003.   DOI   ScienceOn
15 Hirose E., Ohtake S. I. and Azumi K.: Morphological characterization of the tunic in the edible ascidian, Halocynthia roretzi (Drasche), with remarks on 'soft tunic syndrome' in aquaculture. J. Fish Dis., 32:433-445, 2009   DOI   ScienceOn
16 Kumagai A., Suto A., Ito H., Tanabe T., Takahashi K., Kamaishi T. and Miwa S.: Mass mortality of cultured ascidians Halocynthia roretzi associated with softening of the tunic and flagellate-like cells. Dis. Aquat. Organ., 90:223-234, 2010.   DOI   ScienceOn
17 Kumagai A., Suto A., Ito H., Tanabe T., Song J. Y., Kitamura S. I., Hirose E., Kamaishi T. and Miwa S.: Soft tunic syndrome in the edible ascidian Halocynthia roretzi is caused by a kinetoplastid protist. Dis. Aquat. Organ., 95:153-161, 2011.   DOI   ScienceOn
18 Mansour N. R. and Bickle Q. D.: Comparison of microscopy and alamar blue reduction in a larval based assay for schistosome drug screening. PLoS Negl. Trop. Dis. 4: e795, 2010.   DOI   ScienceOn
19 Nociari M. M., Shalev A., Benias P. and Russo C.: A novel one-step, highly sensitive fluorometric assay to evaluate cell-mediated cytotoxicity. J. Immunol. methods., 213:157-167, 1998.   DOI   ScienceOn