Fig 1.
Perkinsus marinus trophozoites grown on agar plates.
Cell size of P. marinus trophozoites plated on plates containing variable percentage of agar.
Fig 2.
Perkinsus spp. colonies on agar plates.
A. General appearance of P. marinus colonies and lawn growing on agar plates (60 mm x 15 mm). B. Close up of square on A showing colonies and P. marinus lawn. C. General appearance of P. mediterraneus colonies after streaking the cultures on agar plates, note individual colonies (arrowheads). D. Low magnification of P. marinus colonies (arrowheads). E. Detail of P. marinus schizogony; note the daughter cells being released from the mother cell (arrowheads). F. Large P. marinus colony; note that cell division is not synchronized as indicated by the differences in cell size.
Fig 3.
Perkinsus mediterraneus MOE[MOE]: GFP cloning on agar plates.
A. Perkinsus mediterraneus MOE[MOE]: GFP growing in clumps in liquid medium (bright field) B. Blue light excitation; note the mother cell cell-wall (arrowheads). C. Detail of P. mediterraneus MOE[MOE]: GFP and non-fluorescent P. mediterraneus growing as single colonies after spreading the culture on the plate (bright field). D. Blue light excitation.
Fig 4.
Effect of triclosan on plated Perkinsus marinus five days after exposure.
A. Overview of the plates. B. Detail of the cells on the plates (40x).
Fig 5.
Possible applications of plating Perkinsus marinus.
A. Perkinsus sp. Isolation. Tissue samples (e.g. hemolymph) or filtrates from waters close to bivalve aquaculture operations could be directly deposited on the plates. B. Subcloning of P. marinus isolates or transfectants expressing fluorescent proteins by spreading the diluted sample on the plate or by streaking (this study). C. Phenotyping Perkinsus spp. and strains based on the colony morphology. D. Mutagenesis. Perkinsus sp. culture is exposed to mutant agents and plates infused with specific inhibitor/substrates for selection depending on the nature of the mutant phenotype of interest or based on cell/colony morphology. E. Extracellular product analysis. The solid medium can be infused with specific substrates to analyze and compare their degradation as the Perkinsus sp. colony grows over time. F. Tropism analysis by depositing a Perkinsus sp. on the center of the plate and components of the bivalve host in different parts of the plate. G. Perkinsus sp. virus isolation. Perkinsus sp. culture supernatant or filtrates from waters close to bivalve aquaculture areas could be directly deposited on Perkinsus sp. lawn plates, incubate, and monitor the plates for formation of lysis plaques.