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 coral Pocillopora damicornis and Vibrio coralliilyticus is commensal. An increase in temperature, or in the abundance of V. coralliilyticus, can turn this association pathogenic, causing tissue lysis, expulsion of the corals' symbiotic algae (genus Symbiodinium), and eventually coral death. Using a combination of microfluidics, fluorescence microscopy, stable isotopes, electron microscopy and NanoSIMS isotopic imaging, we provide insights into the onset and progression of V. coralliilyticus infection in the daytime and at night, at the tissue and (sub-)cellular level. The objective of our study was to connect the macro-scale behavioural response of the coral to the micro-scale nutritional interactions that occur between the host and its symbiont. In the daytime, polyps enhanced their mucus production, and actively spewed pathogens. Vibrio infection primarily resulted in the formation of tissue lesions in the coenosarc. NanoSIMS analysis revealed infection reduced 13C-assimilation in Symbiodinium, but increased 13C-assimilation in the host. In the night incubations, no mucus spewing was observed, and a mucus film was formed on the coral surface. Vibrio inoculation and infection at night showed reduced 13C-turnover in Symbiodinium, but did not impact host 13C-turnover. Our results show that both the nutritional interactions that occur between the two symbiotic partners and the behavioural response of the host organism play key roles in determining the progression and severity of host-pathogen interactions. More generally, our approach provides a new means of studying interactions (ranging from behavioural to metabolic scales) between partners involved in complex holobiont systems, under both homoeostatic and pathogenic conditions.

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

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.

Nutrient enrichment caused by in situ fish farms at Eilat, Red Sea is detrimental to coral reproduction.

Recent studies report conflicting results concerning the effects of eutrophication on coral reproduction. The present study examines reproductive effort in the brooding coral Stylophora pistillata exposed to chronic eutrophication caused by in situ fish cages (FC) in the northern Gulf of Eilat (Aqaba). Histological studies of 20 S. pistillata colonies transplanted to each of two study sites, one close to the nutrient enriched FC site and the other at a reference site (IUI), 8 km southwest of the FC site, show that, overall, corals from the FC site have a significantly higher percentage of polyps containing oocytes and testes than corals from the IUI site. However, average oocyte size and the percentage of oocytes reaching the size at which fertilization occurs (i.e., >200 microm) were both significantly greater in colonies at the IUI site compared to the FC site. As the reproductive season progressed, colonies at the IUI site exhibited a decrease in the percentage of polyps containing oocytes, concomitant with an increase in the number of polyps containing planulae, indicating successful development of oocytes into planulae. In contrast, in colonies at the FC site oocyte numbers were greatest at the end of the reproductive season, and overall, numbers of planulae were significantly lower compared with the IUI colonies, suggesting relative failure of oocyte maturation, fertilization and ensuing larval development. The significantly higher lipid content found during the reproduction season in IUI colonies compared with FC colonies corroborates this assertion. This data strongly suggest that nutrients released from the fish farms have adverse effects on successful production of larvae of S. pistillata. In view of the recent severe deterioration of the coral reefs of Eilat and their present critical state of health, the only chance for their renewal is the use of immediate, prudent and rational protection measures against all man-made perturbations.