SPECIES DISCRIMINATION OF VIBRIO HARVEY CLADE PRIMERS, AND THE FIRST REPORT ON V. HARVEYI AND V. CAMPBELLII IN SOUTHEASTERN U.S.A. WATERS**

Authors

Karagan X. Royer, Georgia College and State University, Milledgeville, GA 31061Follow
Cameron E. Brown, Clemson University, Clemson, SC 29634
Andrei I. Barkovskii, Georgia College and State University, Milledgeville, GA 31061Follow

Abstract

In our recent study, a wide array of virulence genes carried by Vibrio Harveyi clade pathogens has been detected in Georgia aquaculture waters and sediments. Identification of their carriers was attempted with qPCR and a set of previously published species-specific primers. Based on their described selectivity, the following primers were selected and used. Vibrio alginolyticus was targeted with gyrB gene set of primers, and V. parahaemolyticus was targeted with toxR gene set of primers. Due to a lack of primers distinguishing between Vibrio harveyi and Vibrio campbellii, both species were targeted simultaneously with rpoA gene set of primers. Related to the clade Vibrio splendidus was targeted with 16S rRNA gene set of primers. The set of primers for V. splendidus strongly cross-reacted with all species. The set of primers for V. parahaemolyticus cross-reacted with V. alginolyticus and V. campbellii. Previously, these primers have been reported as discriminating between species of Harveyi clade. Our results call for more careful testing of published species-specific primers and qPCR parameters; and possibly for re-evaluation of published data obtained with the above primers by purchasing new primers. The set of primers for gyrB in V. alginolyticus, and for rpoA gene in V. harveyi and V. campbellii exhibited clear discrimination between Vibrio Harvey clade species. No cross-reactions with any other Harveyi clade species or negative control were observed. With the rpoA set of primers, V. harveyi and V.campbelli were detected in water and sediments collected from clams (Mercenaria mercenaria) aquaculture and reefs of wild eastern oysters (Crassostrea virginica). Either one or both of these Vibrio species were detected from June through October with much higher concentrations in sediments. This is the first report on V. harveyi and V. campbellii pathogens in Southeastern USA waters which indicates that Vibrio species are expanding to new territories.

Recommended Citation

Royer, Karagan X.; Brown, Cameron E.; and Barkovskii, Andrei I. (2024) "SPECIES DISCRIMINATION OF VIBRIO HARVEY CLADE PRIMERS, AND THE FIRST REPORT ON V. HARVEYI AND V. CAMPBELLII IN SOUTHEASTERN U.S.A. WATERS**," Georgia Journal of Science, Vol. 82, No. 1, Article 37.
Available at: https://digitalcommons.gaacademy.org/gjs/vol82/iss1/37