Screening for probiotics of Greenshell™ mussel larvae, Perna canaliculus, using a larval challenge bioassay

A bioassay was developed to screen and select bacterial strains as probiotics for an alternative to antibiotic usage in the management of bacterial pathogens of Greenshell™ mussel (GSM) larvae, Perna canaliculus. Sixty-nine isolates originating from a GSM hatchery environment were tested for probiotic activity in larval pathogen-challenge bioassays conducted in tissue culture dishes (TCDs). Vibrio splendidus and a V. coralliilyticus/neptunius-like isolate, Vibrio sp. DO1, were the tested pathogens. Forty of the tested isolates afforded larval survival significantly greater than pathogen controls (p < 0.05). The bioassay technique achieved a 58% success rate in searching for putative probiotics and highlighted the benefit of including the host animal in the first stage of the screening procedure. The time of inoculation of putative probiotic strains prior to pathogen challenge influenced the outcome of the assay. A pre-exposure period of 20 h revealed a greater number of potential probiotics than a two-hour pre-exposure period. Pilot challenge tests, under normal hatchery conditions, confirmed the usefulness of the TCD screening method in recognising effective probiotics.