Coral bleaching, i.e. the loss of photosynthetic algal symbionts, caused by ocean warming is now the main factor driving reef decline, but not all corals are affected equally. Corals from the Arabian Seas have unusually high temperature tolerances, and recently studies implicated salinity as one of the contributing factors. In particular, a recent heat stress experiment at different salinities using the model system Aiptasia and Red Sea corals, showed that cnidaria at large bleach less at heat stress under high salinities and that this is associated with an increase of the osmolyte, floridoside Here we were interested to assess microbial community changes under heat stress at different salinity levels and whether this could help to explain the increase in thermal tolerance of the metaorganism at high salinities. We determined microbial community composition via HiSeq 16S rRNA gene amplicon sequencing of two anemone strains that differ in their associated symbionts, namely H2-SSB01 (type B1) and CC7-SSA01 (type A4), after six days under ambient (25 °C) and heat stress (34 °C) temperatures at salinities of 36, 39, and 42. Both anemones harbored distinct microbial communities, irrespective of temperature or salinity, that were also different from the bacteria in surrounding seawater. Within both host-endosymbiont pairings, the bacterial community composition at low (36) and intermediate (39) salinities did not differ between ambient and heat stress, but was significantly different at high (42) salinities. Subsequent elucidation of bacterial indicator species revealed several taxa that could be associated with a response to temperature and salinity. Our results underline that microbial community composition adjusts under different environmental settings. Importantly, microbial community dynamics of H2-SSB01 aligned with observed differences in bleaching susceptibility and thermal tolerance, whereas the pattern remains unclear for CC7-SSA01, which harbors an intrinsically higher thermal tolerance. Such responses could argue for a contribution of the microbiome to the observed increase in temperature tolerance of the Aiptasia metaorganism at increased salinities. An alternative interpretation is that the microbiome changes denotes a parallel response to changing salinities.

Coral reefs are biologically and economically important ecosystems underpinned by corals that are able to flourish in oligotrophic waters due to their mutualistic association with dinoflagellate symbionts (genus Symbiodinium). Symbiodinium are strictly intracellular, residing within the gastrodermal tissues of the coral host, and contributing the majority of the coral’s energy requirements. Coral reefs are in rapid decline due to a range of threats such as local human influences, bleaching (loss of Symbiodnium and/or reduction of pigment), disease and ocean acidification, to which links to climate change have been made. The close association of corals and a diverse community of microbes led to development of the coral holobiont hypothesis, in which a range of microorganisms (e.g Bacteria) form a functionally-relevant mutualistic relationship with corals and Symbiodinium. This thesis aimed to fill knowledge gaps in the coral holobiont hypothesis and the host-microbe interactions within this system, including pathogen interactions and coral immune system functioning. This thesis revealed that host-microbe interactions in corals are complex, and that the underlying mechanisms of immunity and symbiosis may be similar. The findings corroborate the idea that corals maintain specific bacterial communities that have potential probiotic and nutritional value. In particular, a group of common coral associates were identified, and it is suggested that members of this group are globally occurring key associates. Corals affected by a disease previously described as “White Syndrome” were observed to undergo pronounced changes in their microbial community structure in comparison to healthy colonies. However, in contrast to previous findings, no single pathogen could be identified as the causative agent of the disease syndrome, and it is speculated that corals experiencing altered health status result in a breakdown of the resident associated microbial community structure. Culturable bacterial isolates from corals were shown to affect the growth of each other and in particular some species had great inhibitory properties. Hence, the presence of some bacterial species has the potential to influence the all over structure of the coral associated microbial community. It was also shown that changed environmental conditions may alter the growth conditions for coral associated bacteria in mucus. It is suggested that increased replication is needed in studies of bacterial assemblages on corals, as variability between coral species and sites were observed. In addition, studies of the role of coral microbial communities in health and disease should broaden their focus to more thoroughly consider the role of the coral holobiont, especially with regards to the coral host. This thesis identified the first functional Pattern Recognition Protein (PRP), a C-type lectin named Millectin, in scleractinian corals. Millectin was isolated by affinity chromatography and was shown to bind to bacterial pathogens as well as coral Symbiodinium symbionts. Gene expression of Millectin was upregulated in response to immune stimuli and the lectin was further abundantly expressed in the tissues of corals, suggesting a major role for this protein in system functioning and immunity. Further research into Millectin and a complement factor C3 homolog suggested that these molecules may have been co-opted into the equally important role of symbiont recruitment. Gene expression analysis of C3 also indicated this molecule may be involved in responses to tissue trauma. Millectin shows variability in the binding region, and hence, is the earliest evolutionary representative to date of a variable PRP. This finding, and the observed ancestral relation with vertebrate homologs, provided further information on the evolution of the innate immune system and gives further insight into invertebrate immunity.

Coral reefs are under considerable decline. The framework builders in coral reefs are scleractinian corals, which comprise so-called holobionts, consisting of cnidarian host, algal symbionts (genus Symbiodinium), and other associated microbes. Corals are commonly considered stenohaline osmoconformers, possessing limited capability to adjust to salinity changes. However, corals differ in their ability to cope with different salinities. The underlying mechanisms have not yet been addressed. To further understand putative mechanisms involved, I examined coral holobiont osmoregulation conducting a range of experiments on the coral Fungia granulosa. In my research F. granulosa from the Red Sea exhibited pronounced physiological reactions (decreased photosynthesis, cessation of calcification) upon short-term incubations (4 h) to high salinity (55). However, during a 29-day in situ salinity transect experiment, coral holobiont photosynthesis was unimpaired under high salinity (49) indicating acclimatization. F. granulosa microbiome changes after the 29-day high salinity exposure aligned with a bacterial community restructuring that putatively supports the coral salinity acclimatization (osmolyte synthesis, nutrient fixation/cycling). Long-term incubations (7 d) of cultured Symbiodinium exhibited cell growth even at ‘extreme’ salinity levels of 25 and 55. Metabolic profiles of four Symbiodinium strains exposed to increased (55) and decreased (25) salinities for 4 h indicated distinct carbohydrates and amino acids to be putatively involved in the osmoadjustment. Importantly, under high salinity the osmolyte floridoside was consistently increased. This could be corroborated in the coral model Aiptasia and in corals from the Persian/Arabian Gulf, where floridoside was also markedly increased upon short- (15 h) and long-term (>24 months) exposure to high salinity, confirming an important role of floridoside in the osmoadjustment of cnidarian holobionts. This thesis demonstrates osmoacclimatization of F. granulosa and osmoadjustment of cultured Symbiodinium. All three main compartments (i.e. coral host, Symbiodinium, bacteria) seem to contribute to the coral holobionts salinity adjustment. However, the exact mechanisms of coral host and bacteria contribution remain to be determined. Floridoside likely constitutes a conserved osmolyte increasing the salinity resilience of Symbiodinium and also of the cnidarian/coral holobiont. Floridoside further possess’ antioxidative properties, possibly providing a protection from reactive oxygen species formation as a result of salinity stress or/and other environmental stressors.