Diagnostic tools for rapid detection and quantification of Weissella ceti NC36 infections in rainbow trout
| Autor: | Timothy J. Welch, Jeffrey M. Hinshaw, A.K. Snyder |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Ecology
Outbreak Brain Weissella ceti Biology Diagnostic tools 16S ribosomal RNA Real-Time Polymerase Chain Reaction Applied Microbiology and Biotechnology Rapid detection Sensitivity and Specificity Bacterial Load Microbiology Disease Outbreaks Fish Diseases Oncorhynchus mykiss RNA Ribosomal 16S Weissella North Carolina Animals Rainbow trout Pathogen load Pathogen Gram-Positive Bacterial Infections Spleen |
| Zdroj: | Letters in applied microbiologyReferences. 60(2) |
| ISSN: | 1472-765X |
| Popis: | Weissellosis of rainbow trout is caused by the Gram-positive bacteria Weissella ceti and has been reported in China, Brazil and the United States. This disease can result in high mortality in market-sized fish and thus can cause significant economic loss. Thus far, phenotypic characterization and 16S rRNA sequencing have been used to confirm a Weissellosis diagnosis. Here, we present the development of PCR-based diagnostic tools for the rapid identification and quantification of W. ceti within bacteriological culture and infected tissues. A duplex PCR, which amplifies both genus- and strain-specific targets, positively identifies isolates as W. ceti NC36. A qPCR assay was also developed to quantify pathogen load from infected tissues, using a W. ceti NC36 unique locus. A proof of concept study was performed to demonstrate that quantification using traditional plate count methods and qPCR were significantly correlated when assessed from infected brain and spleen tissue. These tools were also used to confirm diagnosis of Weissellosis in a commercial rainbow trout farm during an outbreak investigation. These are the first diagnostic tools developed for identification and quantification of W. ceti infection within rainbow trout, contributing to rapid Weissellosis diagnosis, enhanced pathogen surveillance and epidemiological studies. Significance and Impact of the Study Weissellosis is a rapidly emerging infectious disease of farmed rainbow trout caused by Weissella ceti, a Gram-positive lactic-acid bacterium. This disease can cause significant economic loss, and thus, efforts to limit its spread are urgently needed and are dependent on the development of diagnostic tools. The article presents the development of PCR-based diagnostic assays for the rapid identification of W. ceti and for quantification of pathogen load directly from tissues without the need for bacterial amplification or isolation. These tools are critical for rapid confirmation of Weissellosis diagnosis and for studying the epidemiology of this emerging pathogen. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text