Edinburgh Napier University

  Benthic microbial communities can play an important role in the induction of larval metamorphosis in marine invertebrates. The role of biofilms in recognizing the parental habitat is unknown for the ecologically and economically important mangrove crab Ucides cordatus, a species suffering from environmental pressures such as habitat degradation and disease. In the laboratory, we experimentally tested the influence of biofilms grown in offshore and estuarine waters on the moulting rates and developmental time to metamorphosis (TTM) of the last larval stage (megalopa) of U. cordatus. Here, we specifically studied whether: (i) megalopae of this larval-exporting species respond differently to marine (MB) and estuarine biofilms (EB), (ii) biofilms previously immersed (24 h) in adult crab-conditioned seawater (ACSW; i.e. conspecific chemical cues) exhibit a greater effectiveness (i.e. by inducing higher moulting rates and shorter TTM) on larval metamorphosis than non-immersed ones and (iii) biofilms pre-immersed (24 h) in ACSW decrease their effectiveness when incubated (24 h) in pure filtered seawater (FSW) before offering them to megalopae. U. cordatus megalopae metamorphosed to the first juvenile crab stage (JI) in response to both MB and EB, however the moulting rates were significantly higher and development TTM shorter in the presence of EB compared to MB. The metamorphosis-inducing effects were significantly enhanced when both types of biofilms were pre-immersed in ACSW, suggesting that conspecific cues can be absorbed and stored by microbial matrix. The higher inductiveness of EB compared to MB persisted after previous contact with ACSW. Furthermore, EB exhibited a significantly greater biomass production (measured as dry weight) than MB, and when immersed in ACSW, both groups of biofilms significantly increased in biomass (maintaining the hierarchy EB > MB), suggesting that water-soluble chemical substances emitted by the adult crabs may have been absorbed and metabolized by the biofilms. The metamorphic inductiveness and biomass production decreased when both groups of biofilms that were previously kept (24 h) in ACSW were thereafter immersed (24 h) in FSW. All megalopae successfully moulted to JI after accelerated TTM when reared in ACSW, regardless of presence or absence of biofilms, corroborating that conspecific stimuli are the most effective metamorphosis-stimulating cues tested so far in U. cordatus. The fact that EB improved the moulting rates and shortened the development TTM indicates that megalopae can recognize and respond to microbial assemblages typical for specific environments. This should be ecologically important during larval recruitment by facilitating settlement in appropriate habitats for the post-metamorphic development of early benthic recruits. In addition, the influence of EB could encourage colonization of new estuarine areas and aid natural recovery of U. cordatus in mangrove habitats where crab populations have suffered significant reduction due to deforestation, fishing pressure or diseases.