A history of the international conferences on coelenterate biology

From the author's opening address as President of the Fifth International Conference on Coelenterate Biology, given at Southampton University, 9 July 1989.     

Effects of disturbance on coral communities: bleaching in Moorea,French Polynesia

This study examines patterns of susceptibility and short-term recovery of corals from bleaching. A mass coral bleaching event began in March, 1991 on reefs in Moorea, French Polynesia and affected corals on the shallow barrier reef and to >20 m depth on the outer forereef slope. There were significant differences in the effect of the bleaching among common coral genera, with Acropora, Montastrea, Montipora, and Pocillopora more affected than Porites, Pavona, leptastrea or Millepora. Individual colonies of the common species of Acropora and Pocillopora were marked and their fate assessed on a subsequent survey in August, 1991 to determine rates of recovery and mortality. Ninety-six percent of Acropora spp. showed some degree of bleaching compared to 76% of Pocillopora spp. From March to August mortality of bleached colonies of Pocillopora was 17%, 38% recovered completely, and many suffered some partial mortality of the tissue. In contrast, 63% of the Acropora spp. died, and about 10% recovered completely. Generally, those colonies with less than 50% of the colony area affected by the bleaching recovered at a higher rate than did those with more severe bleaching. Changes in community composition four months after the event began included a significant decrease only in crustose algae and an increase in cover of filamentous algae, much of which occupied plate-like and branching corals that had died in the bleaching event. Total coral cover and cover of susceptible coral genera had declined, but not significantly, after the event.     

Habitat selection and juvenile persistence control the distribution of two closely related Caribbean damselfishes

Summary On many Caribbean fringing coral reefs, two closely related and ecologically similar damselfishes, the beaugregory (Stegastes leucostictus Müller and Troschel) and the cocoa damselfish (S. variabilis Castelnau), occupy nonoverlapping vertical distributions. In St. Croix (USVI), beaugregory are very abundant in shallow water back reef habitats (1–2 m depth) while cocoa damselfish are restricted to the base of the forereef (10–15 m depth).In this study, the roles of habitat selection at settlement and juvenile persistence were investigated to determine their influence on this pattern of zonation. Settlement events observed at intervals over a two-year period revealed that habitat selection occurred at settlement and was confined to habitats occupied by adults. In addition, differences in juvenile persistence (due to mortality and/or emigration) were found when species were translocated between depths. Over a period of 100-days, juvenile beaugregory moved from 1 m to 12 m depth suffered four-fold greater losses at the deeper sites than shallow water controls, while translocated cocoa damselfish suffered twice as many losses in shallow water than controls at 12 m depth. Despite these differences in persistence, growth rates of the two species were similar and independent of depth. These results indicate that preferential habitat selection at settlement, perhaps an evolutionary response to differential juvenile mortality, may play a deciding role in determining distributions of ecologically similar species of coral reef fishes.     

Protein expression and genetic structure of the coral Porites lobata in an environmentally extreme Samoan back reef: does host genotype limit phenotypic plasticity?

The degree to which coral reef ecosystems will be impacted by global climate change depends on regional and local differences in corals’ susceptibility and resilience to environmental stressors. Here, we present data from a reciprocal transplant experiment using the common reef building coral Porites lobata between a highly fluctuating back reef environment that reaches stressful daily extremes, and a more stable, neighbouring forereef. Protein biomarker analyses assessing physiological contributions to stress resistance showed evidence for both fixed and environmental influence on biomarker response. Fixed influences were strongest for ubiquitin‐conjugated proteins with consistently higher levels found in back reef source colonies both pre and post‐transplant when compared with their forereef conspecifics. Additionally, genetic comparisons of back reef and forereef populations revealed significant population structure of both the nuclear ribosomal and mitochondrial genomes of the coral host (F_ST = 0.146 P < 0.0001, F_ST = 0.335 P < 0.0001 for rDNA and mtDNA, respectively), whereas algal endosymbiont populations were genetically indistinguishable between the two sites. We propose that the genotype of the coral host may drive limitations to the physiological responses of these corals when faced with new environmental conditions. This result is important in understanding genotypic and environmental interactions in the coral algal symbiosis and how corals may respond to future environmental changes.     

Mesophotic communities of the insular shelf at Tutuila, American Samoa

An investigation into the insular shelf and submerged banks surrounding Tutuila, American Samoa, was conducted using a towed camera system. Surveys confirmed the presence of zooxanthellate scleractinian coral communities at mesophotic depths (30–110 m). Quantification of video data, separated into 10-m-depth intervals, yielded a vertical, landward-to-seaward and horizontal distribution of benthic assemblages. Hard substrata composed a majority of bottom cover in shallow water, whereas unconsolidated sediments dominated the deep insular shelf and outer reef slopes. Scleractinian coral cover was highest atop mid-shelf patch reefs and on the submerged bank tops in depths of 30–50 m. Macroalgal cover was highest near shore and on reef slopes approaching the bank tops at 50–60 m. Percent cover of scleractinian coral colony morphology revealed a number of trends. Encrusting corals belonging to the genus Montipora were most abundant at shallow depths with cover gradually decreasing as depth increased. Massive corals, such as Porites spp., displayed a similar trend. Percent cover values of plate-like corals formed a normal distribution, with the highest cover observed in the 60–70 m depth range. Shallow plate-like corals belonged mostly to the genus Acropora and appeared to be significantly prevalent on the northeastern and eastern banks. Deeper plate-like corals on the reef slopes were dominated by Leptoseris, Pachyseris, or Montipora genera. Branching coral cover was high in the 80–110 m depth range. Columnar and free-living corals were also occasionally observed from 40–70 m.     

Diversity and spatial pattern of coral communities in the Red Sea upper twilight zone

Summary This is the first study based on numerical analysis of the abundance of 11 scleractinian corals of depths at between 100–210 m in the Red Sea twilight zone. Two distinct coral communities were found: a Leptoseris fragilis community at a depth of 100–130 m (zone 1) and a Dendrophillia horsti community below 130 m (zone 2, 3). Population densities and coral coverage are very low; distribution of individuals is highly clumped. Highest observed densities on 100 m² were 2720 individuals for L. fraglis, 2720 for D. horsti and 2260 for Javania insignis. Calculated coverage rates were maximally 3.6% (L. fragilis), 0.08% (D. horsti) and 0.11% (J. insignis). L. fragilis, the only symbiont bearing coral, was very abundant. It has an unusual depth range for a photosynthesising coral. Coral density is only weakly correlated with hard bottom coverage. Species diversity with an average of 8 species is highest at 120–170 m and decreases in shallower and deeper water. The study depth range is a transient zone for coral distribution. It contains the upper distribution limits of a few deep sea corals and the lower ones of several shallower water species. Ahermatypic corals, collected at 160–170 m depth, were transplanted from their original depth to 159, 118, 70 and 40 m; after one year most species survived transplantation far beyond their upper distributional limits. The symbiotic L. fragilis, collected at 120 m, survived transplantation to deep water (159 m) as well as shallow zones (90, 70 and 40 m). The study demonstrates the feasibility of line-transect methods for coral community studies with a submersible.     

Deepwater broadcast spawning by Montastraea cavernosa, Montastraea franksi, and Diploria strigosa at the Flower Garden Banks, Gulf of Mexico

Broadcast spawning by corals is a tightly synchronized process characterized by co-ordinated gamete release within 30–60 min time windows once per year. In shallow water corals, annual water temperature cycles set the month, lunar periodicity the day, and sunset time the hour of spawning. This tight temporal regulation is critical for achieving high fertilization rates in a pelagic environment. Given the differences in light and temperature that occur with depth and the importance of these parameters in regulating spawn timing, it has been unclear whether deeper coral can respond to the same environmental cues that regulate spawning behaviour in shallower coral. In this report, a remotely operated vehicle was used to monitor coral spawning activity at the Flower Garden Banks at depths from 33 to 45 m. Three species Montastraea cavernosa, Montastraea franksi, and Diploria strigosa were documented spawning within this depth range. All recorded spawning events were within the same temporal windows as shallower conspecifics. These data indicate that deep corals at this location either sense the same environmental parameters, despite local attenuation, or communicate with shallower colonies that can sense such spawning cues.     

Changes in morphology and abundance of Stylophora pistillata and Acropora variabilis (anthozoa,hexactiniaria) with respect to depth at Elat,Red Sea

Entire colonies of Stylophora pistillata and Acropora variabilis were collected along a depth transect 100 m south of the Marine Biological Laboratory of Hebrew University Elat, Israel. The branch height of Acropora variabilis decreases significantly with respect to depth, and this species is equitably distributed from 5 to 30 m. Stylophora pistillata demonstrated no decrease in branch height with respect to depth and its distribution is limited to shallow water. The changes in morphology and distribution enhance the credibility of a suggestion made by Roos (1967) that corals common at all depths would show changes in morphology that minimized self-shading. Stylophora pistillata demonstrated an increase in branch density with respect to depth. A hypothesis, based upon the occurrence of Lithophaga cumingiana and water turbulence, is presented to explain the increase in branch density with respect to depth of Stylophora pistillata.     

Depth-related changes to density,diversity and structure of benthic megafaunal assemblages in the Fimbul ice shelf region,Weddell Sea,Antarctica

The depth-related patterns in the benthic megafauna of the NE Weddell Sea shelf at the edge of the Fimbul Ice Shelf were investigated at seven sites using towed camera platform photographs. Megafaunal density decreased with depth from 77,939 ha⁻¹ at 245 m to 8,895 ha⁻¹ at 510 m. While diversity was variable, with H′ ranging between 1.34 and 2.28, there were no depth related patterns. Multivariate analyses revealed two distinct assemblages; a shallow assemblage with dense patches of suspension feeders in undisturbed areas and a deep assemblage where these were not present. Disturbance from icebergs explained many observed patterns in faunal distribution. In shallow waters probable effects of disturbance were observed as changes in successional stages; in deeper waters changes in habitat as a result of past disturbance explained faunal distributions. In deeper areas ice ploughing created a mosaic landscape of fine and coarse sediments. Total megafaunal density was highest in areas of coarse sediment (up to 2.9 higher than in finer sediment areas) but diversity was highest in intermediate areas (H′ = 2.35).     

In situ release of mucus and DOC-lipid from the corals Acropora variabilis and Stylophora pistillata in different light regimes

Rates of mucus and DOC-lipid release were determined for colonies of Acropora variabilis and Stylophora pistillata at 5 m depth and for a colony of A. variabilis at 23 m depth. In addition, colonies at 5 m were shaded to simulate ambient irradiance at 6 m, 10 m and 16 m depth to evaluate the effect of light on the rates of release. A. variabilis released more mucus and DOC-lipid at 5 m than at 23 m depth. For both corals, the night rates were about 30% those of the day. A reduction in total integrated irradiance decreased mucus output from the corals. Similarly, DOC-lipid release showed a diurnal pattern and diminished with reduction in daily irradiance. For both coral species, DOC-lipid release rates were greater in the afternoon than in the morning. The night rates were less than 55% those of the day. The DOC-lipid comprised wax esters and a phospholipid fraction. The diurnal variation was due to changes in yield of wax esters which contributed >90% of the carbon released as DOC-lipid. In situ release of mucus and DOC-lipid was infuenced by light effects on phototrophic carbon metabolism. A daily budget for carbon released as mucus and DOC-lipid was estimated for each coral species at 5 m depth.     

Geographic differences in vertical connectivity in the Caribbean coral Montastraea cavernosa despite high levels of horizontal connectivity at shallow depths

The deep reef refugia hypothesis proposes that deep reefs can act as local recruitment sources for shallow reefs following disturbance. To test this hypothesis, nine polymorphic DNA microsatellite loci were developed and used to assess vertical connectivity in 583 coral colonies of the Caribbean depth‐generalist coral Montastraea cavernosa. Samples were collected from three depth zones (≤10, 15–20 and ≥25 m) at sites in Florida (within the Upper Keys, Lower Keys and Dry Tortugas), Bermuda, and the U.S. Virgin Islands. Migration rates were estimated to determine the probability of coral larval migration from shallow to deep and from deep to shallow. Finally, algal symbiont (Symbiodinium spp.) diversity and distribution were assessed in a subset of corals to test whether symbiont depth zonation might indicate limited vertical connectivity. Overall, analyses revealed significant genetic differentiation by depth in Florida, but not in Bermuda or the U.S. Virgin Islands, despite high levels of horizontal connectivity between these geographic locations at shallow depths. Within Florida, greater vertical connectivity was observed in the Dry Tortugas compared to the Lower or Upper Keys. However, at all sites, and regardless of the extent of vertical connectivity, migration occurred asymmetrically, with greater likelihood of migration from shallow to intermediate/deep habitats. Finally, most colonies hosted a single Symbiodinium type (C3), ruling out symbiont depth zonation of the dominant symbiont type as a structuring factor. Together, these findings suggest that the potential for shallow reefs to recover from deep‐water refugia in M. cavernosa is location‐specific, varying among and within geographic locations likely as a consequence of local hydrology.     

Carbonate production by scleractinian corals at Aqaba,Gulf of Aqaba,Red Sea

Summary In a fringing reef at Aqaba at the northern end of the Gulf of Aqaba (29°26′N) growth rates, density, and the calcification rate ofPorites were investigated in order to establish calculations of gross carbonate production for the reefs in this area. Colony accretion ofPorites decreases with depth as a function of decreasing growth rates. The calcification rate ofPorites is highest in shallow water (0–5 m depth) with 0.9 g·cm⁻²·yr⁻¹ and falls down to 0.5 g·cm⁻²·yr⁻¹ below 30 m. Scleractinian coral gross production is calculated from potential productivity and coral coverage. It is mainly dependent on living coral cover and to a lesser extent on potential productivity. Total carbonate production on the reef ranged from 0 to 2.7 kg/m² per year, with a reef-wide average of 1.6 kg/m² perycar. Maximum gross carbonate production by corals at Aqaba occurs at the reef crest and in the middle fore-reef from 10 to 15 m water depth. Production is low in sandy reef parts. Below 30 m depth values still reach ca. 50% of shallow water values. Mean potential production of colonies and gross carbonate production of the whole reef community at Aqaba is lower than in tropical reefs. However, carbonate production is higher than in reef areas at the same latitude in the Pacific, indicating a northward shift of reef production in the Red Sea.     

Composition and sources of near reef zooplankton on a Jamaican forereef along with implications for coral feeding

Nocturnal near-reef zooplankton from the forereef of Discovery Bay, Jamaica, were sampled during winter and summer 1994 using a diver-operated plankton pump with an intake head positioned within centimeters of benthic zooplanktivores. The pump collected zooplankton not effectively sampled by conventional net tows or demersal traps. We found consistently greater densities of zooplankton than did earlier studies that used other sampling methods in similar locations. There was no significant difference between winter (3491±578 m^–3) and summer (2853±293 m^–3) zooplankton densities. Both oceanic- and reef-associated forms were found at temporal and spatial scales relevant to benthic suspension feeders. Copepods were always the most abundant group, averaging 89% of the total zooplankton, and most were not of demersal origin. The cyclopoids, Oithona spp., were the numerically dominant organisms, with an average density of 1684±260 m^–3. Other zooplankton (e.g., shrimp larvae, crab larvae, polychaetes, chaetognaths, amphipods, and isopods) were highly variable and much less abundant. Near-reef zooplankton abundances were high throughout the night sampling period, not just after sunset and before sunrise as previously described. Mean biomass was 4.5 mg C m^–3, with values ranging from 1.0 to 15.6 mg C m^–3. This work has important implications for evaluating which zooplankton types are available to benthic suspension feeders, including corals.     

New family of sea anemones (Actiniaria,Acontiaria) from deep polar seas

We describe and illustrate two new species from polar deep seas that belong to a new genus and family. Antipodactidae fam. nov. is characterized by acontia with macrobasic p-amastigophores; this type of nematocyst has never been reported from acontia. Antipodactis gen. nov. is characterized by a column with a distinct scapus and scapulus, cuticle-bearing tenaculi on the scapus, more mesenteries proximally than distally, mesenteries regularly arranged, restricted and reniform retractor musculature, and macrobasic p-amastigophores in the acontia. Antipodactis scotiae sp. nov. and A. awii sp. nov. differ in number of mesenteries, retractor and parietobasilar muscles, cnidae, and geographic distribution. We discuss the familial and generic characters of Antipodactis gen. nov. and its relationship to other families of acontiarian sea anemones: it most closely resembles members of Kadosactidae in terms of anatomy and some aspects of cnidom, and has a cnidom identical to that of Diadumenidae in terms of the types of nematocysts. Because the morphology of nematocysts is critical to the diagnosis of this family, we review and comment on the nomenclature of mastigophores. The macrobasic p-amastigophores of Antipodactidae fam. nov. conform to England’s (Hydrobiologia 216/217:691–697, 1991) definition rather than that of Mariscal (Coelenterate Biology. Academic Press, New York, pp 129–178, 1974).     

Symbiosis-specific changes in dimethylsulphoniopropionate concentrations in Stylophora pistillata along a depth gradient

Scleractinian corals are prolific producers of dimethylsulphoniopropionate (DMSP), but ecophysiological mechanisms influencing cellular concentrations are uncertain. While DMSP is often proposed to function as an antioxidant, interactions between specific host–symbiont genotype associations, plasticity in DMSP concentrations and environmental conditions that can either exert or alleviate oxidative stress are unclear. We used long-term (6 months) reciprocal transplantation of Stylophora pistillata hosting two distinct symbiont phylotypes along a depth gradient, clades A (<20 m) and C (>20 m), to assess the effect of change in depth (light intensity) on DMSP concentrations in relation to symbiont genotype and photoacclimation in corals between 3 and 50 m in the Gulf of Aqaba. Bathymetric distribution of total DMSP (DMSPt) per cell varied significantly while particulate DMSP (DMSPp) appeared to be unaffected by depth. Highest DMSPt concentrations in control corals occurred at 20 m. While 3-m transplants showed a significant increase in DMSPt concentration at 20 m and became affiliated with an additional genotype (C72), 50-m transplants largely persisted with their original genotype and exhibited no significant changes in DMSPt concentrations. DMSPt concentrations in transplants at both 3 and 50 m, on the other hand, increased significantly while all corals maintained their original symbiont genotypes. Photoacclimation differed significantly with transplantation direction relative to the controls. Symbionts in 3-m transplants at 20 m exhibited no changes in chlorophyll a (chl a) concentration, cell density or cell diameter while symbiont densities decreased and chl a concentrations increased significantly at 50 m. In contrast, symbiont densities in 50-m transplants remained unaffected across depths while symbiont diameters decreased. Chl a concentrations decreased at 20 m and increased at 3 m. Our results indicate that DMSPt concentrations following changes in depth are not only a function of symbiont genotype but result from different acclimation abilities of both symbiotic partners.

     

Damage to the leeward reefs of Curaçao and Bonaire,Netherlands Antilles from a rare storm event: Hurricane Lenny,November 1999

Fringing reefs along the southwestern shores of the Caribbean islands of Curaçao and Bonaire (12°N), located outside the most frequent hurricane tracks, are rarely affected by heavy wave-action and major storms, yet have experienced disturbances such as coral bleaching, coral diseases, and mass mortalities. The last major hurricane to hit these islands occurred over 100 yr ago. In November 1999, Hurricane Lenny took an unusual west-to-east track, bisecting the Caribbean Basin and passing approximately 200 miles north of Curaçao and Bonaire. The leeward shores of both islands were pounded for 24 h by heavy waves (~3–6 m) generated while the storm was centered far to the west. Reef damage surveys at 33 sites conducted between November 1999 to April 2000, following the storm, documented occurrences of toppling, fragmentation, tissue damage, bleaching, and smothering due to the storm. Reefs were severely damaged along westward-facing shores but less impacted where the reef front was tangential to the wave direction or was protected by offshore islands. At the most severely damaged sites, massive coral colonies 2–3-m high (older than 100 yr) were toppled or overturned, smaller corals were broken loose and tumbled across the shallow reef platform and either deposited on the shore or dropped onto the deeper forereef slope. Branching and plating growth forms suffered more damage than massive species and large colonies experienced greater damage than small colonies. Toppled massive corals have a high potential of preserving the event signature even if they survive and continue to grow. Reorientation of large, long-lived coralla may provide a unique indicator of disturbance in a reef system rarely affected by hurricanes. At some locations, wave scouring removed loose sediment to reveal a cemented framework of Acropora cervicornis rubble on the shallow platform above 10-m depth. This rubble was generated in situ, not by storm processes, but rather by an earlier mass mortality of thickets of staghorn coral that covered extensive areas of the shallow platform prior to the incidence of white band disease in the early 1980s.     

Phenotypic plasticity for skeletal growth,density and calcification of <Emphasis Type="Italic">Porites lobata</Emphasis> in response to habitat type

A reciprocal transplant experiment (RTE) of the reef-building coral Porites lobata between shallow (1.5 m at low tide) back reef and forereef habitats on Ofu and Olosega Islands, American Samoa, resulted in phenotypic plasticity for skeletal characteristics. Transplants from each source population (back reef and forereef) had higher skeletal growth rates, lower bulk densities, and higher calcification rates on the back reef than on the forereef. Mean annual skeletal extension rates, mean bulk densities, and mean annual calcification rates of RTE groups were 2.6–9.8 mm year⁻¹, 1.41–1.44 g cm⁻³, and 0.37–1.39 g cm⁻² year⁻¹ on the back reef, and 1.2–4.2 mm year⁻¹, 1.49–1.53 g cm⁻³, and 0.19–0.63 g cm⁻² year⁻¹ on the forereef, respectively. Bulk densities were especially responsive to habitat type, with densities of transplants increasing on the high energy forereef, and decreasing on the low energy back reef. Skeletal growth and calcification rates were also influenced by source population, even though zooxanthella genotype of source colonies did not vary between sites, and there was a transplant site x source population interaction for upward linear extension. Genetic differentiation may explain the source population effects, or the experiment may have been too brief for phenotypic plasticity of all skeletal characteristics to be fully expressed. Phenotypic plasticity for skeletal characteristics likely enables P. lobata colonies to assume the most suitable shape and density for a wide range of coral reef habitats.     

The role of zoobenthos in energy flow in two shallow lakes

Net production of zoobenthos in two shallow and eutrophic lakes, i.e. the S-basin of Mývatn, Iceland (maximum depth 4.2 m, mean depth 2.3 m) and Hjarbæk Fjord, Denmark (maximum depth 6.5 m, mean depth 1.9 m) were calculated as 878 and 1093 kJ m^-2 yr^-1, respectively. The zoobenthos in both lakes was dominated by Chironomidae (Diptera) living partly as filtrators feeding on suspended particles (phytoplankton) and partly as surface feeders foraging on benthic algae and/or seston. Respiration and consumption were estimated from the literature. Net production efficiency averaged 0.41 and 0.48 in Hjarbæk Fjord and Mývatn, respectively. Ingestion was dominated by herbivorous chironomids, while detritivorous tubificids were insignificant. Zoobenthic production made up 86% of total secondary production (zooplankton plus zoobenthos) in both lakes. The trophic efficiency between net primary production and benthic net secondary production was 8% and 11% in Hjarbæk Fjord and Lake Mývatn S-basin, respectively.     

Out of Their Depth? Isolated Deep Populations of the Cosmopolitan Coral Desmophyllum dianthus May Be Highly Vulnerable to Environmental Change

Deep sea scleractinian corals will be particularly vulnerable to the effects of climate change, facing loss of up to 70% of their habitat as the Aragonite Saturation Horizon (below which corals are unable to form calcium carbonate skeletons) rises. Persistence of deep sea scleractinian corals will therefore rely on the ability of larvae to disperse to, and colonise, suitable shallow-water habitat. We used DNA sequence data of the internal transcribed spacer (ITS), the mitochondrial ribosomal subunit (16S) and mitochondrial control region (MtC) to determine levels of gene flow both within and among populations of the deep sea coral Desmophyllum dianthus in SE Australia, New Zealand and Chile to assess the ability of corals to disperse into different regions and habitats. We found significant genetic subdivision among the three widely separated geographic regions consistent with isolation and limited contemporary gene flow. Furthermore, corals from different depth strata (shallow <600 m, mid 1000-1500 m, deep >1500 m) even on the same or nearby seamounts were strongly differentiated, indicating limited vertical larval dispersal. Genetic differentiation with depth is consistent with the stratification of the Subantarctic Mode Water, Antarctic Intermediate Water, the Circumpolar Deep and North Pacific Deep Waters in the Southern Ocean, and we propose that coral larvae will be retained within, and rarely migrate among, these water masses. The apparent absence of vertical larval dispersal suggests deep populations of D. dianthus are unlikely to colonise shallow water as the aragonite saturation horizon rises and deep waters become uninhabitable. Similarly, assumptions that deep populations will act as refuges for shallow populations that are impacted by activities such as fishing or mining are also unlikely to hold true. Clearly future environmental management strategies must consider both regional and depth-related isolation of deep-sea coral populations.