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Vibrio owensii  DY05 is a serious pathogen causing epizootics in the larviculture of ornate spiny lobster  Panulirus ornatus . In the present study a multi-tiered probiotic screening strategy was used to identify a probiotic combination capable of protecting  P. ornatus  larvae (phyllosomas) from experimental  V. owensii  DY05 infection. From a pool of more than 500 marine bacterial isolates, 91 showed definitive  in vitro  antagonistic activity towards the pathogen. Antagonistic candidates were shortlisted based on phylogeny, strength of antagonistic activity, and isolate origin. Miniaturized assays used a green fluorescent protein labelled transconjugant of  V. owensii  DY05 to assess pathogen growth and biofilm formation in the presence of shortlisted candidates. This approach enabled rapid processing and selection of candidates to be tested in a phyllosoma infection model. When used in combination, strains  Vibrio  sp. PP05 and  Pseudoalteromonas  sp. PP107 significantly and reproducibly protected  P. ornatus  phyllosomas during vectored challenge with  V. owensii  DY05, with survival not differing significantly from unchallenged controls. The present study has shown the value of multispecies probiotic treatment and demonstrated that natural microbial communities associated with wild phyllosomas and zooplankton prey support antagonistic bacteria capable of  in vivo  suppression of a pathogen causing epizootics in phyllosoma culture systems.

Identification of an Antagonistic Probiotic Combination Protecting Ornate Spiny Lobster (Panulirus ornatus) Larvae against Vibrio owensii Infection