Felicia Soth

Friday class 1:00 PM

11/4/2013

First Record of Renibacterium Salmoninarum In The Sea Lamprey

Introduction: Renibacterium is a bacterial disease that causes Bacterial Kidney Disease. It is often found widespread throughout the Great Lakes. It greatly affects salmonine fish. It was believed that only salmonines were carriers, however, recent studies have proven that the Pacific Hake and the Pacific Herring may be carriers as well. Many non-native fish have invaded the Great Lakes causing the numbers of native fish to drop as well as causing great economic loss. Reasons for study: The purpose of this study was to determine if F. salmoninarum was jumping from salmonine fish to other species (specifically the sea lamprey). This study took place near the Great Lakes. Because so much loss, economically and ecologically, occurred due to the Sea Lamprey the Great Lakes Fishery Commission began collecting male Lampreys and making them sterile, then releasing them into the St. Marys River. This was to cause less female lampreys to get fertilized eggs, reducing their numbers. But this constant shifting of fish from place to place could very easily spread different kinds of pathogens around. Methods and materials: In 2003, some adult Lamprey male and females in spawning-phase were retrieved from the Humber River and Duffins Creek and were then given alive to the Aquatic Animal Health Laboratory. The fish were then euthanized with an excess amount of MS 222. They were then dissected and striped of their kidneys. Their kidneys were homogenized and spread onto MKDM plates. These plates were incubated for 20 days. Isolates that grew were then tested using nPCR. Tissue pellets were then obtained by centrifugation. The controls and products were loaded into the wells of a 2% agarose gel for gel electrophoresis. The gel was placed in a 1X Tris gel buffer inside of an electrophoresis chamber. The samples were to be considered positive of R. salmoninarum if they had a band and 320 base pairs.

Felicia Soth

Friday class 1:00 PM

11/4/2013

Results: R. salmoninarum was isolated from the collected sea lampreys extracted kidneys. It was not however found to be in the tested blood samples. Every isolate was tested to be gram-positive diplobacilli or coccobacilli. The colonies on the plates were white, shiny, smooth, had round colonies, and they were about 1mm in diameter. All retrieved isolates were confirmed with nPCR and Q-ELISA. With nPCR R. salmoninarum was detected in 66% of the kidneys of lampreys collected from Duffins creek and 5% from the Humber River. Discussion: This was the first report of the R. salmoninarum in the sea lamprey. Even though it was previously believed to be a salmonid-specific pathogen, it was found in sea lampreys from Lake Ontario for two consecutive years. While R. salmoninarum was found in the sea lampreys kidneys, it was not found to be in any other internal organ nor the blood. The isolates that had been found in the sea lampreys had the same attraction to kidneys as the R. salmoninarum found in salmonids. The size of the isolates found in the sea lampreys was the same size as what was published for R. salmoninarum. Tissues form the isolates retrieved was not consistent with the results. Conclusion: The reason for the tissues not being consistent may have been from bile salts that accumulated in the sea lampreys muscles and kidneys due to the fact that they have no gall bladder. Bile salts act like a detergent and may have been the reason for the inhibition on diagnostic assays. The once thought to be salmonine-specific R. salmoninarum is now beginning to jump species. In previous studies there had been no reports of the sea lamprey carrying R. salmoninarum. With this study there is proof that sea lampreys have some amounts of this disease. The sea lamprey can now be reported as a host of Renibacterium Salmoninarum. Limitations of this study: Due to the fact that the Sea Lamprey does not have a gall bladder makes it hard to perform diagnostic assays. This is because without a gall bladder bile salts build up within the muscles and kidneys which can cause an inhibition to nPCR and Q-ELISA. http://www.jwildlifedis.org/doi/pdf/10.7589/0090-3558-42.3.556

Felicia Soth

Friday class 1:00 PM

11/4/2013