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Veterinary Parasitology 183 (2011) 8–13

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Veterinary Parasitology
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Antiparasitic efficacy of dihydrosanguinarine and
dihydrochelerythrine from Macleaya microcarpa against
Ichthyophthirius multifiliis in richadsin (Squaliobarbus curriculus)
Jia-yun Yao a , Zhi-ming Zhou a , Xi-lian Li b , Wen-linYin a , Hong-shun Ru a , Xiao-yi Pan a ,
Gui-jie Hao a , Yang Xu a , Jin-yu Shen a,∗

Zhejiang Institute of Freshwater Fisheries, Huzhou, Zhejiang 313001, China
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China



Article history:
Received 15 January 2011
Received in revised form 21 June 2011
Accepted 8 July 2011
Keywords:Ichthyophthirius multifiliis

Ichthyophthirius multifiliis is a holotrichous protozoan that invades the gills and skin surfaces of fish and can cause morbidity and high mortality in most species of freshwater
fish worldwide. The present study was undertaken to investigate the antiparasitic activity of crude extracts and purecompounds from the leaves of Macleaya microcarpa. The
chloroform extract showed a promising antiparasitic activity against I. multifiliis. Based on
these finding, the chloroform extract was fractionated on silica gel column chromatography in a bioactivity-guided isolation affording two compounds showing potent activity. The
structures of the two compounds were elucidated as dihydrosanguinarine anddihydrochelerythrine by hydrogen and carbon-13 nuclear magnetic resonance spectrum and electron
ionization mass spectrometry. The in vivo tests revealed that dihydrosanguinarine and dihydrochelerythrine were effective against I. multifiliis with median effective concentration
(EC50 ) values of 5.18 and 9.43 mg/l, respectively. The acute toxicities (LC50 ) of dihydrosanguinarine anddihydrochelerythrine for richadsin were 13.3 and 18.2 mg/l, respectively.
The overall results provided important information for the potential application of dihydrosanguinarine and dihydrochelerythrine in the therapy of serious infection caused by I.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction
The ciliate Ichthyophthirius multifiiis is one of the most
pathogenic parasites offish, and it constitutes a major
disease problem in aquaculture (Bisharyan et al., 2003;
Dickerson and Dawe, 1995; McCallum, 1985, 1986). The
morbidity rate due to this disease may reach up to 100%,
causing great economic losses in aquaculture and ornamental fish breeding (Osman et al., 2009).

∗ Corresponding author. Tel.: +86 572 2045132; fax: +86 572 2041403.
E-mail (J.-y. Yao), (J.-y. Shen).
0304-4017/$ – see front matter © 2011 Elsevier B.V. All rights reserved.

To cope with the parasitism and deleterious consequences, various parasiticides have been used in
aquaculture. Formerly, ichthyophthiriasis was treated
effectively with malachite green, a compound that has been
banned on fish farms due toits confirmed carcinogenicity in most countries (Alderman, 1985; Wahli et al., 1993).
Since the ban, formalin (Stoskopf, 1993; Rowland et al.,
2009), copper sulfate (Straus et al., 2009; Rowland et al.,
2009), potassium permanganate (Straus and Griffin, 2001;
Buchmann et al., 2003), hydrogen peroxide (Lahnsteiner
and Weismann, 2007), sodium percarbonate (Heinecke and
Buchmann, 2009) have beenused with varying levels of
success. Strategic treatments using sodium percarbonate in
combination with continuous filtration of the pond water

J.-y. Yao et al. / Veterinary Parasitology 183 (2011) 8–13

are suggested to provide a powerful and more environmentally friendly method to minimize and control infections
with I. multifiliis. In terms of current strategies for controlling...
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