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DOI: 10.3852/09-027
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Mycologia, 102(2), 2010, pp. 478-485.
© 2010 by The Mycological Society of America

Microwell enumeration of viable Saprolegniaceae in water samples


Even Thoen

     National Veterinary Institute, PO Box 750 Sentrum, N-0106 Oslo, Norway

Øystein Evensen

     Norwegian School of Veterinary Science, PO Box 8146 Dep., N-0033 Oslo, Norway

Ida Skaar 1

     National Veterinary Institute, PO Box 750 Sentrum, N-0106 Oslo, Norway

Existing methods for enumeration of viable Saprolegniaceae propagules in water are laborious, time consuming and prevent examination of large numbers of samples or samples with high spore loads. In the present study a microwell plate (MWP) assay for estimation of Saprolegniaceae in water samples, modified from Hagen (1992), was evaluated. The ability of the assay to recover Saprolegniaceae was assessed by applying it to spore suspensions with four predetermined concentrations, 500–10 000 spores per liter of samples tested. The method also was used to analyze a set of field samples and compare it to a standard filtration method to ascertain its practicability. The MWP assay underestimated the number of spores in the test suspensions with predetermined concentrations. The accuracy of the assay varied with spore concentration, giving the lowest recovery (62.5%) at low spore numbers and the highest (86%) at intermediate concentrations (1000–5000 spores/L) for both isolates and growth media. The findings indicate that spores aggregate with increasing concentration. When applied to field samples the assay clearly distinguished among samples with presumptive differences in spore load and yielded significantly higher counts than the filtration method. The results justify the MWP method for use in estimation of Saprolegniaceae in water bodies particularly relevant for monitoring of spore load in aquaculture as well as in ecological studies.

Key words: quantification method, Saprolegniaceae, spores, water


1 Corresponding author. E-mail: ida.skaar{at}vetinst.no







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