Detection and Distribution of Flavobacterium columnare in Experimentally-Infected Channel Catfish, Ictalurus punctatus Using Culture and Polymerase Chain Reaction

Abstract

The authors examined the use of culture and polymerase chain reaction (PCR) to detect Flavobacterium columnare in experimentally-infected channel catfish, Ictalurus punctatus. Five treatments were utilized which included immersion exposure to 10⁶, 10⁷, 10⁸ colony forming units (CFU)/mL for 30 minutes, intramuscular injection of 10⁸ CFU/fish and a negative control (i.e., immersion in Cytophaga broth). Flavobacterium columnare was isolated and detected in mucus 30 minutes following exposure by microbiological culture and PCR in all treatments except the negative controls. Gills were positive by culture and by PCR in all treatments at 30 minutes post treatment except the 10⁶ CFU/mL immersion treatment which did not yield positive culture and PCR results until 1 hour. Culture positive samples were observed in the internal organs (anterior and posterior kidney) and blood of the 10⁷⁸ CFU/mL treatments although at low numbers (≤10 CFU). Results of PCR paralleled that of culture for the mucus and gill samples when analyzing all treatments together over time suggesting either method is useful in determination of the presence ofF. columnare. Polymerase chain reaction was significantly (P < 0.001) better at detection ofF. columnare from skin/muscle than was the use of microbiological culture. These results suggest that PCR may be useful for rapid detection of F. columnare in the mucus.

Key Words:

• Flavobacterium columnare
• PCR
• channel catfish
• columnaris disease
• Ictalurus punctatus