FLV tracer assays
Prepare FLVs for tracer assays .
Collect seawater samples from the desired location .
After determining the concentration of the FLVs in the concentrate , and the ambient concentration of viruses in the seawater , the proper amount of FLV concentrate should be added at tracer levels ( < 10 % of original ambient virus concentration ) into sample volumes of no less than 400 mL .
Designate a control treatment .
At each time point , total viral abundance and FLV numbers should be determined in duplicate from small volume subsamples ( 5–30 mL ) taken into sterile , 15 - or 50 - mL polyethylene tubes .
Subsamples are immediately fixed with 1 % to 2 % ( final concentration ) 0.02 - µm filtered formalin and stored at 4°C .
Slides should be prepared according to Noble and Fuhrman ( 1998 ) or Patel et al . ( 2007 ) .
Production and removal rates are calculated from the equations of Glibert et al . ( 1982 ) and Fuhrman ( 1987 ) ( see guidelines for equations ) .
