Membrane filtration – Fluorescent antibody staining procedure for detecting and quantifying Renibacterium salmoninarum in coelomic fluid of Chinook Salmon (Oncorhynchus tshawytscha)
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Abstract
We developed a rapid method for detecting and quantifying the pathogen Renibacterium salmoninarum in coelomic fluid of spring chinook salmon (Oncorhynchus tshawytscha) by concentrating the bacteria on 0.2-μm polycarbonate filters and staining them with specific fluorescein-labeled antibody. Centrifugation of samples and resuspension of the sedimented material in phosphate-buffered saline containing Triton X-100 increased the ease of filtration. Background fluorescence was reduced by counterstaining filters with Eriochrome black T. Postfiltration staining, rinsing, and counterstaining were done in the syringe-mounted filter holders, reducing handling of the filters and possible loss of bacteria. The number of bacteria detected by the filtration – fluorescent antibody technique in a broth culture of R. salmoninarum ranged from 6.7 × 107to7.6 × 107/mL and was slightly higher than that determined by plate count (9.6 × 106/mL). Increasing the sample dilution or decreasing the number of microscope fields examined generally increased the variability of filter counts of R. salmoninarum. Using the filtration – fluorescent antibody technique, we detected the bacterium in the coelomic fluid of 85% of spawning female spring chinook salmon sampled from a hatchery population.
| Publication type | Article |
|---|---|
| Publication Subtype | Journal Article |
| Title | Membrane filtration – Fluorescent antibody staining procedure for detecting and quantifying Renibacterium salmoninarum in coelomic fluid of Chinook Salmon (Oncorhynchus tshawytscha) |
| Series title | Canadian Journal of Fisheries and Aquatic Sciences |
| DOI | 10.1139/f87-027 |
| Volume | 44 |
| Issue | 1 |
| Year Published | 1987 |
| Language | English |
| Publisher | NRC Research Press |
| Contributing office(s) | Western Fisheries Research Center |
| Description | 5 p. |
| First page | 206 |
| Last page | 210 |
| Google Analytic Metrics | Metrics page |