Vibrio coralliilyticus infection triggers a behavioural response and perturbs nutritional exchange and tissue integrity in a symbiotic coral

Under homoeostatic conditions, the relationship between the coralPocillopora damicornisandVibrio coralliilyticusiscommensal. An increase in temperature, or in the abundance ofV. coralliilyticus, can turn this association pathogenic,causing tissue lysis, expulsion of the corals’symbiotic algae (genusSymbiodinium), and eventually coral death. Using acombination of microfluidics,fluorescence microscopy, stable isotopes, electron microscopy and NanoSIMS isotopicimaging, we provide insights into the onset and progression ofV.coralliilyticusinfection in the daytime and at night, atthe tissue and (sub-)cellular level. The objective of our study was to connect the macro-scale behavioural response ofthe coral to the micro-scale nutritional interactions that occur between the host and its symbiont. In the daytime, polypsenhanced their mucus production, and actively spewed pathogens.Vibrioinfection primarily resulted in the formationof tissue lesions in the coenosarc. NanoSIMS analysis revealed infection reduced13C-assimilation inSymbiodinium, butincreased13C-assimilation in the host. In the night incubations, no mucus spewing was observed, and a mucusfilm wasformed on the coral surface.Vibrioinoculation and infection at night showed reduced13C-turnover inSymbiodinium, but didnot impact host13C-turnover. Our results show that both the nutritional interactions that occur between the two symbioticpartners and the behavioural response of the host organism play key roles in determining the progression and severity ofhost-pathogen interactions. More generally, our approach provides a new means of studying interactions (ranging frombehavioural to metabolic scales) between partners involved in complex holobiont systems, under both homoeostatic andpathogenic conditions

Using NanoSIMS coupled with microfluidics to visualize the early stages of coral infection by Vibrio coralliilyticus

Abstract Background Global warming has triggered an increase in the prevalence and severity of coral disease, yet little is known about coral/pathogen interactions in the early stages of infection. The point of entry of the pathogen and the route that they take once inside the polyp is currently unknown, as is the coral’s capacity to respond to infection. To address these questions, we developed a novel method that combines stable isotope labelling and microfluidics with transmission electron microscopy (TEM) and nanoscale secondary ion mass spectrometry (NanoSIMS), to monitor the infection process between Pocillopora damicornis and Vibrio coralliilyticus under elevated temperature. Results Three coral fragments were inoculated with 15N-labeled V. coralliilyticus and then fixed at 2.5, 6 and 22 h post-inoculation (hpi) according to the virulence of the infection. Correlative TEM/NanoSIMS imaging was subsequently used to visualize the penetration and dispersal of V. coralliilyticus and their degradation or secretion products. Most of the V. coralliilyticus cells we observed were located in the oral epidermis of the fragment that experienced the most virulent infection (2.5 hpi). In some cases, these bacteria were enclosed within electron dense host-derived intracellular vesicles. 15N-enriched pathogen-derived breakdown products were visible in all tissue layers of the coral polyp (oral epidermis, oral gastrodermis, aboral gastrodermis), at all time points, although the relative 15N-enrichment depended on the time at which the corals were fixed. Tissues in the mesentery filaments had the highest density of 15N-enriched hotspots, suggesting these tissues act as a “collection and digestion” site for pathogenic bacteria. Closer examination of the sub-cellular structures associated with these 15N-hotspots revealed these to be host phagosomal and secretory cells/vesicles. Conclusions This study provides a novel method for tracking bacterial infection dynamics at the levels of the tissue and single cell and takes the first steps towards understanding the complexities of infection at the microscale, which is a crucial step towards understanding how corals will fare under global warming

Metabolic fluxes and chemical signaling during coral disease at the single cell-level.

Metabolic fluxes and chemical signaling during coral disease at the single cell-level

Additional file 6: of Using NanoSIMS coupled with microfluidics to visualize the early stages of coral infection by Vibrio coralliilyticus

Correlative TEM/NanoSIMS in the infected fragment fixed at 22 h. (PDF 170 kb