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Quantitative PCR detection of Batrachochytrium dendrobatidis DNA from sediments and water

Diseases of Aquatic Organisms

By:
, , , ,
DOI: 10.3354/dao01831

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Abstract

The fungal pathogen Batrachochytrium dendrobatidis (Bd) causes chytridiomycosis, a disease implicated in amphibian declines on 5 continents. Polymerase chain reaction (PCR) primer sets exist with which amphibians can be tested for this disease, and advances in sampling techniques allow non-invasive testing of animals. We developed filtering and PCR based quantitative methods by modifying existing PCR assays to detect Bd DNA in water and sediments, without the need for testing amphibians; we tested the methods at 4 field sites. The SYBR based assay using Boyle primers (SYBR/Boyle assay) and the Taqman based assay using Wood primers performed similarly with samples generated in the laboratory (Bd spiked filters), but the SYBR/Boyle assay detected Bd DNA in more field samples. We detected Bd DNA in water from 3 of 4 sites tested, including one pond historically negative for chytridiomycosis. Zoospore equivalents in sampled water ranged from 19 to 454 l-1 (nominal detection limit is 10 DNA copies, or about 0.06 zoospore). We did not detect DNA of Bd from sediments collected at any sites. Our filtering and amplification methods provide a new tool to investigate critical aspects of Bd in the environment. ?? Inter-Research 2007.

Additional Publication Details

Publication type:
Article
Publication Subtype:
Journal Article
Title:
Quantitative PCR detection of Batrachochytrium dendrobatidis DNA from sediments and water
Series title:
Diseases of Aquatic Organisms
DOI:
10.3354/dao01831
Volume
77
Issue:
1
Year Published:
2007
Language:
English
Larger Work Type:
Article
Larger Work Subtype:
Journal Article
First page:
11
Last page:
15
Number of Pages:
5