Sven Uthicke

Changes in coral-associated microbial communities during a bleaching event

Environmental stressors such as increased sea surface temperatures are well-known for contributing to coral bleaching; however, the effect of increased temperatures and subsequent bleaching on coral-associated microbial communities is poorly understood. Colonies of the hard coral Acropora millepora were tagged on a reef flat off Magnetic Island (Great Barrier Reef) and surveyed over 2.5 years, which included a severe bleaching event in January/February 2002. Daily average water temperatures exceeded the previous 10-year average by more than 1 °C for extended periods with field-based visual surveys recording all tagged colonies displaying signs of bleaching. During the bleaching period, direct counts of coral zooxanthellae densities decreased by ∼64%, before recovery to pre-bleaching levels after the thermal stress event. A subset of three tagged coral colonies were sampled through the bleaching event and changes in the microbial community elucidated. Denaturing gradient gel electrophoresis (DGGE) analysis demonstrated conserved bacterial banding profiles between the three coral colonies, confirming previous studies highlighting specific microbial associations. As coral colonies bleached, the microbial community shifted and redundancy analysis (RDA) of DGGE banding patterns revealed a correlation of increasing temperature with the appearance of Vibrio-affiliated sequences. Interestingly, this shift to a Vibrio-dominated community commenced prior to visual signs of bleaching. Clone libraries hybridized with Vibrio-specific oligonucleotide probes confirmed an increase in the fraction of Vibrio-affiliated clones during the bleaching period. Post bleaching, the coral microbial associations again shifted, returning to a profile similar to the fingerprints prior to bleaching. This provided further evidence for corals selecting and shaping their microbial partners. For non-bleached samples, a close association with Spongiobacter-related sequences were revealed by both clone libraries and DGGE profiling. Despite Vibrio species being previously implicated in bleaching of specific coral species, it is unsure if the relative increase in retrieved Vibrio sequences is due to bacterial infection or an opportunistic response to compromised health and changing environmental parameters of the coral host. This study provides the first molecular-based study demonstrating changes in coral-associated bacterial assemblages during a bleaching event on a natural reef system.

A boom–bust phylum? Ecological and evolutionary consequences of density variations in echinoderms

Echinoderms play a key role in structuring many marine ecosystems and are notorious for large population density variations in so-called “outbreak” or “dieoff” events. In a review of this phenomenon, we assess the causal factors and ecological and evolutionary consequences. We identified 28 species (6 Asteroidea, 8 Echinoidea, 10 Holothuroidea, 4 Ophiuroidea) that exhibit large (more than two population doublings or halvings) population density changes. Three generalized patterns were identified and named for exemplary species: (1) rapid decreases followed by no or slow recovery (Diadema–Paracentrotus Model), (2), rapid increase and apparent stability at a new population density (Amperima–Amphiura Model), and (3) population density fluctuations (Acanthaster–Asterias Model). Echinoderms identified were distributed from the shallow intertidal to the deep sea, and from tropical to temperate regions. In most cases, significant impacts on the respective ecosystems were observed. The most striking similarity among all species identified was possession of the ancestral-type planktotrophic larva. This larval type was significantly overrepresented in species identified within the Asteroidea, Echinoidea, Holothuroidea, and for the combined data set. We suggest three main factors that render a life history with planktotrophic larvae a high-risk–high-gain strategy: (1) a strong nonlinear dependency of larval production on adult densities (Allee effects), (2) a low potential for compensatory feedback mechanisms, and (3) an uncoupling of larval and adult ecology. The alternative (derived) lecithotrophic larva occurs in 68% of recent echinoderm species, suggesting an evolutionary trend toward this larval type. Lecithotrophic development represents a more buffered life history because compensatory feedback between adult densities and larval output is likely to be more efficient. For lecithotrophic developers, direct nutritive coupling from adult to larva to the early benthic juvenile provides a buffer against starvation. Lecithotrophic larvae are independent of the vagaries of planktonic food supply, and their short planktonic duration may promote local recruitment. Anthropogenic influences contributed to the population density variations in most cases, including increased primary productivity through eutrophication or global change, disease, overfishing, and species introductions. We suggest that anthropogenic disturbance, through its influence on the frequency and/or amplitude of echinoderm population density changes, may go beyond present ecosystem impacts and alter future evolutionary trends.

Ecological effects of ocean acidification and habitat complexity on reef-associated macroinvertebrate communities

The ecological effects of ocean acidification (OA) from rising atmospheric carbon dioxide (CO2) on benthic marine communities are largely unknown. We investigated in situ the consequences of long-term exposure to high CO2 on coral-reef-associated macroinvertebrate communities around three shallow volcanic CO2 seeps in Papua New Guinea. The densities of many groups and the number of taxa (classes and phyla) of macroinvertebrates were significantly reduced at elevated CO2 (425–1100 µatm) compared with control sites. However, sensitivities of some groups, including decapod crustaceans, ascidians and several echinoderms, contrasted with predictions of their physiological CO2 tolerances derived from laboratory experiments. High CO2 reduced the availability of structurally complex corals that are essential refugia for many reef-associated macroinvertebrates. This loss of habitat complexity was also associated with losses in many macroinvertebrate groups, especially predation-prone mobile taxa, including crustaceans and crinoids. The transition from living to dead coral as substratum and habitat further altered macroinvertebrate communities, with far more taxa losing than gaining in numbers. Our study shows that indirect ecological effects of OA (reduced habitat complexity) will complement its direct physiological effects and together with the loss of coral cover through climate change will severely affect macroinvertebrate communities in coral reefs.

Gene flow and population history in high dispersal marine invertebrates: mitochondrial DNA analysis of Holothuria nobilis (Echinodermata: Holothuroidea) populations from the Indo-Pacific

The sea cucumber, Holothuria nobilis, has a long‐lived planktotrophic larvae, and previous allozyme surveys have suggested that high dispersal is realized. In contrast, recent ecological studies indicate that dispersal is low. To reconcile these data, and to investigate the evolution of this Indo‐Pacific species, we screened geographical variation in 559 bp of a mitochondrial gene (COI) in 360 samples from the Australasian region and La Réunion. Sequences from La Réunion differed by > 7% from others and may constitute another species. Haplotype diversity in other samples was high (0.942, SD = 0.007), but haplotypes were closely related (mean nucleotide diversity: 0.0075, SD = 0.0041). amova, pairwise FST values and exact tests did not detect significant population structure. Nested clade analysis showed that one of two main clades was over‐represented in west Australia, whereas the other was more common in the northern Great Barrier Reef. Isolation‐by‐distance was identified as the main determinant of population structure at several clade levels. Contiguous range expansion was inferred for evolutionary older clade levels and this may correspond to a late Pleistocene (88 000–193 000 years ago) population expansion inferred from haplotype mismatch distributions. Thus, the population genetic structures detected are likely to be formed prior to the last ice age, with some indications for high dispersal on shorter time scales.

Effects of ocean acidification on microbial community composition of, and oxygen fluxes through, biofilms from the Great Barrier Reef

Rising anthropogenic CO(2) emissions acidify the oceans, and cause changes to seawater carbon chemistry. Bacterial biofilm communities reflect environmental disturbances and may rapidly respond to ocean acidification. This study investigates community composition and activity responses to experimental ocean acidification in biofilms from the Australian Great Barrier Reef. Natural biofilms grown on glass slides were exposed for 11 d to four controlled pCO(2) concentrations representing the following scenarios: A) pre-industrial (∼300 ppm), B) present-day (∼400 ppm), C) mid century (∼560 ppm) and D) late century (∼1140 ppm). Terminal restriction fragment length polymorphism and clone library analyses of 16S rRNA genes revealed CO(2) -correlated bacterial community shifts between treatments A, B and D. Observed bacterial community shifts were driven by decreases in the relative abundance of Alphaproteobacteria and increases of Flavobacteriales (Bacteroidetes) at increased CO(2) concentrations, indicating pH sensitivity of specific bacterial groups. Elevated pCO(2) (C + D) shifted biofilm algal communities and significantly increased C and N contents, yet O(2) fluxes, measured using in light and dark incubations, remained unchanged. Our findings suggest that bacterial biofilm communities rapidly adapt and reorganize in response to high pCO(2) to maintain activity such as oxygen production.

Natural volcanic CO2 seeps reveal future trajectories for host-microbial associations in corals and sponges

Atmospheric carbon dioxide (CO2) levels are rapidly rising causing an increase in the partial pressure of CO2 (pCO2) in the ocean and a reduction in pH known as ocean acidification (OA). Natural volcanic seeps in Papua New Guinea expel 99% pure CO2 and thereby offer a unique opportunity to explore the effects of OA in situ. The corals Acropora millepora and Porites cylindrica were less abundant and hosted significantly different microbial communities at the CO2 seep than at nearby control sites <500 m away. A primary driver of microbial differences in A. millepora was a 50% reduction of symbiotic Endozoicomonas. This loss of symbiotic taxa from corals at the CO2 seep highlights a potential hurdle for corals to overcome if they are to adapt to and survive OA. In contrast, the two sponges Coelocarteria singaporensis and Cinachyra sp. were ∼40-fold more abundant at the seep and hosted a significantly higher relative abundance of Synechococcus than sponges at control sites. The increase in photosynthetic microbes at the seep potentially provides these species with a nutritional benefit and enhanced scope for growth under future climate scenarios (thus, flexibility in symbiosis may lead to a larger niche breadth). The microbial community in the apparently pCO2-sensitive sponge species S. massa was not significantly different between sites. These data show that responses to elevated pCO2 are species-specific and that the stability and flexibility of microbial partnerships may have an important role in shaping and contributing to the fitness and success of some hosts.

Nutrient regeneration by abundant coral reef holothurians

To investigate quantity and quality of nutrients regenerated by sediment feeding holothurians excretion experiments were conducted with the common species Holothuria atra and Stichopus chloronotus on the Great Barrier Reef (GBR). These experiments were supplemented with measurements of ammonium concentrations behind holothurians in situ, and in the respiratory trees of the animals. For both H. atra and S. chloronotus, ammonium is the only important N-excretion product. Ammonium excretion rates were higher in the latter species and they were higher in summer than in winter for both species. Small amounts of phosphate were also released by both holothurian species. Area-specific nitrogen (0.52–5.35 mg m−2 day−1) and phosphorus (0.01–0.47 mg m−2 day−1) regeneration rates of the holothurians were in a similar range as previously reported nutrient fluxes on coral reefs. Ammonium excretion rates were reduced in products of asexual reproduction (transverse fission) of S. chloronotus, but not in those of H. atra. The fact that anterior sections of both species (which do not have respiratory trees) have normal (H. atra) or only slightly reduced (S. chloronotus) excretion rates, indicates that intact organisms also excrete ammonium through the bodywall. Compared to ambient levels in the water column, ammonium concentrations were higher in the respiratory trees (8–15 μM elevation) and directly behind (1.3–2.4 μM elevation) holothurians when water expulsions through the anus occur. Ammonium excretion through the bodywall in direct contact with benthic microalgae and elevated levels in the respiratory water ejected in close proximity to the sediment are suggested as important mechanisms to retain nutrients in the benthic recycling system.

The stunting effect of a high CO2 ocean on calcification and development in sea urchin larvae, a synthesis from the tropics to the poles

The stunting effect of ocean acidification on development of calcifying invertebrate larvae has emerged as a significant effect of global change. We assessed the arm growth response of sea urchin echinoplutei, here used as a proxy of larval calcification, to increased seawater acidity/pCO2 and decreased carbonate mineral saturation in a global synthesis of data from 15 species. Phylogenetic relatedness did not influence the observed patterns. Regardless of habitat or latitude, ocean acidification impedes larval growth with a negative relationship between arm length and increased acidity/pCO2 and decreased carbonate mineral saturation. In multiple linear regression models incorporating these highly correlated parameters, pCO2 exerted the greatest influence on decreased arm growth in the global dataset and also in the data subsets for polar and subtidal species. Thus, reduced growth appears largely driven by organism hypercapnia. For tropical species, decreased carbonate mineral saturation was most important. No single parameter played a dominant role in arm size reduction in the temperate species. For intertidal species, the models were equivocal. Levels of acidification causing a significant (approx. 10–20+%) reduction in arm growth varied between species. In 13 species, reduction in length of arms and supporting skeletal rods was evident in larvae reared in near-future (pCO2 800+ µatm) conditions, whereas greater acidification (pCO2 1000+ µatm) reduced growth in all species. Although multi-stressor studies are few, when temperature is added to the stressor mix, near-future warming can reduce the negative effect of acidification on larval growth. Broadly speaking, responses of larvae from across world regions showed similar trends despite disparate phylogeny, environments and ecology. Larval success may be the bottleneck for species success with flow-on effects for sea urchin populations and marine ecosystems.

Coral reef invertebrate microbiomes correlate with the presence of photosymbionts

Coral reefs provide habitat for an array of marine invertebrates that host symbiotic microbiomes. Photosynthetic symbionts including Symbiodinium dinoflagellates and diatoms potentially influence the diversity of their host-associated microbiomes by releasing carbon-containing photosynthates and other organic compounds that fuel microbial metabolism. Here we used 16S ribosomal RNA (rRNA) gene amplicon pyrosequencing to characterise the microbiomes of 11 common Great Barrier Reef marine invertebrate species that host photosynthetic symbionts and five taxa in which they are absent. The presence of photosynthetic symbionts influenced the composition but not the species richness, evenness and phylogenetic diversity of invertebrate-associated microbiomes. Invertebrates without photosynthetic symbionts were dominated by Alphaproteobacteria, whereas those hosting photosynthetic symbionts were dominated by Gammaproteobacteria. Interestingly, many microbial species from photosymbiont-bearing invertebrates, including Oceanospirillales spp., Alteromonas spp., Pseudomonas spp., Halomonas spp., are implicated in the metabolism of dimethylsulfoniopropionate (DMSP). DMSP is produced in high concentrations by photosynthetic dinoflagellates and is involved in climate regulation by facilitating cloud formation. Microbiomes correlated with host taxa and replicate individuals from most sampled species grouped in distance-based redundancy analysis of retrieved 16S rRNA gene sequences. This study highlights the complex nature of invertebrate holobionts and confirms the importance of photosynthetic symbionts in structuring marine invertebrate bacterial communities.

Sediment patch selectivity in tropical sea cucumbers (Holothurioidea: Aspidochirotida) analysed with multiple choice experiments

To investigate patch selectivity in aspidochirotide holothurians, individuals of five species (Holothuria (Halodeima) atra Jager, H. (H.) edulis Lesson, H. (Microthele) nobilis Selenka, Stichopus chloronotus Brandt and S. variegatus Semper) were subjected to multiple choice experiments. As a food source, sediments were pre-cultivated in petri dishes under different light and nutrient regimes. This resulted in four sediment treatments with different levels of microalgal biomass (measured as chlorophyll a and phaeophytin concentrations). Only two sediment treatments were used for experiments with H. nobilis and S. variegatus. The sediments were offered simultaneously to individual holothurians (six per experiment), and the weights of the sediment in each petri dish at the start and after 48 h were used to calculate a selection index together with confidence intervals for each food type. In experiments with H. atra and H. edulis, the animals exhibited no preference for any food type. In contrast, S. chloronotus significantly selected sediments with the highest contents of microalgae and avoided the sediment with the lowest pigment concentrations. These results were supported by field collections of sediments found directly underneath holothurians. Sediment underneath H. edulis did not differ from the average sediment of the habitat, while H. atra was found on sediments only slightly higher in chlorophyll a. Chlorophyll a concentrations underneath S. chloronotus were distinctly higher than in the adjacent sediment and that underneath H. atra. H. nobilis showed only a weak preference for sediments with higher pigment concentrations in aquarium experiments, and no patch selectivity in this species was found in the field. Stichopus variegatus exhibited a very distinct patch selectivity towards sediments with more nutritional value in both aquaria experiments and field measurements. Thus members of the genus Holothuria had no, or only a weak, tendency to select their food source, whereas both Stichopus species appeared to carefully select the sediment patch to feed on.