The reproductive biology and early life ecology of a common Caribbean brain coral, <Emphasis Type="Italic">Diploria labyrinthiformis</Emphasis> (Scleractinia: Faviinae)

Despite the fact that most of the severe demographic bottlenecks in coral populations occur during their earliest life stages, information on the reproductive biology and early life history traits of many coral species is limited and often inferred from adult traits only. This study reports on several atypical aspects of the reproductive biology and early life ecology of the grooved brain coral, Diploria labyrinthiformis (Linnaeus, 1758), a conspicuous reef-building species on Caribbean reefs. The timing of gamete release of D. labyrinthiformis was monitored in Curaçao over eight consecutive months, and embryogenesis, planulae behavior, and settlement rates were observed and quantified. We further studied growth and symbiont acquisition in juvenile D. labyrinthiformis for 3.5 yr and compared settler survival under ambient and nutrient-enriched conditions in situ. Notably, D. labyrinthiformis reproduced during daylight hours in six consecutive monthly spawning events between May and September 2013, with a peak in June. This is the largest number of reproductive events per year ever observed in a broadcast-spawning Caribbean coral species. In settlement experiments, D. labyrinthiformis planulae swam to the bottom of culture containers 13 h after spawning and rapidly settled when provided with settlement cues (42% within 14 h). After 5 months, the survival and growth rates of settled juveniles were 3.7 and 1.9 times higher, respectively, for settlers that acquired zooxanthellae within 1 month after settlement, compared to those that acquired symbionts later on. Nutrient enrichment increased settler survival fourfold, but only for settlers that had acquired symbionts within 1 month after settlement. With at least six reproductive events per year, a short planktonic larval phase, high settlement rates, and a positive response to nutrient enrichment, the broadcast-spawning species D. labyrinthiformis displays a range of reproductive and early life-history traits that are more often associated with brooding coral species, illustrating that classical divisions of coral species by reproductive mode alone do not always reflect the true biology and ecology of their earliest life stages.     

Variability in chemical defense across a shallow to mesophotic depth gradient in the Caribbean sponge <Emphasis Type="Italic">Plakortis angulospiculatus</Emphasis>

The transition between shallow and mesophotic coral reef communities in the tropics is characterized by a significant gradient in abiotic and biotic conditions that could result in potential trade-offs in energy allocation. The mesophotic reefs in the Bahamas and the Cayman Islands have a rich sponge fauna with significantly greater percent cover of sponges than in their respective shallow reef communities, but relatively low numbers of spongivores. Plakortis angulospiculatus, a common sponge species that spans the depth gradient from shallow to mesophotic reefs in the Caribbean, regenerates faster following predation and invests more energy in protein synthesis at mesophotic depths compared to shallow reef conspecifics. However, since P. angulospiculatus from mesophotic reefs typically contain lower concentrations of chemical feeding deterrents, they are not able to defend new tissue from predation as efficiently as conspecifics from shallow reefs. Nonetheless, following exposure to predators on shallow reefs, transplanted P. angulospiculatus from mesophotic depths developed chemical deterrence to predatory fishes. A survey of bioactive extracts indicated that a specific defensive metabolite, plakortide F, varied in concentration with depth, producing altered deterrence between shallow and mesophotic reef P. angulospiculatus. Different selective pressures in shallow and mesophotic habitats have resulted in phenotypic plasticity within this sponge species that is manifested in variable chemical defense and tissue regeneration at wound sites.     

Modeling vertical coral connectivity and mesophotic refugia

Whether mesophotic reefs will behave as refugia for corals threatened by global climate change and coastal development depends on vertical exchange of larvae between diverse habitats. Here we use a biophysical model of larval dispersal to estimate vertical connectivity of a broadcasting (Orbicella faveolata) and a brooding (Porites astreoides) species of coral in the US Virgin Islands. Modeling predicts subsidy to shallow areas by mesophotic larvae of both species based on local hydrology, adult reproductive characteristics, larval traits, and a wide range of scenarios developed to test depth-sensitive factors, such as fertilization rates and post-settlement survivorship. In extreme model scenarios of reduced fertilization and post-settlement survivorship of mesophotic larvae, 1–10 % local mesophotic subsidy to shallow recruitment is predicted for both species, which are demographically significant. Although direct vertical connectivity is higher for the broadcaster, the brooder demonstrates higher local multigenerational vertical connectivity, which suggests that local P. astreoides populations are more resilient than those of O. faveolata, and corroborates field studies. As shallow habitat degrades, mesophotic–shallow subsidy is predicted to increase for both species. This study is the first of its kind to simulate larval dispersal and settlement between habitats of different depths, and these findings have local, regional, and global implications for predicting and managing coral reef persistence in a changing climate.     

Modeled connectivity of <Emphasis Type="Italic">Acropora millepora</Emphasis> populations from reefs of the Spratly Islands and the greater South China Sea

The Spratly Island archipelago is a remote network of coral reefs and islands in the South China Sea that is a likely source of coral larvae to the greater region, but about which little is known. Using a particle-tracking model driven by oceanographic data from the Coral Triangle region, we simulated both spring and fall spawning events of Acropora millepora, a common coral species, over a 46-yr period (1960–2005). Simulated population biology of A. millepora included the acquisition and loss of competency, settlement over appropriate benthic habitat, and mortality based on experimental data. The simulations aimed to provide insights into the connectivity of reefs within the Spratly Islands, the settlement of larvae on reefs of the greater South China Sea, and the potential dispersal range of reef organisms from the Spratly Islands. Results suggest that (1) the Spratly Islands may be a significant source of A. millepora larvae for the Palawan reefs (Philippines) and some of the most isolated reefs of the South China Sea; and (2) the relatively isolated western Spratly Islands have limited source reefs supplying them with larvae and fewer of their larvae successfully settling on other reefs. Examination of particle dispersal without biology (settlement and mortality) suggests that larval connectivity is possible throughout the South China Sea and into the Coral Triangle region. Strong differences in the spring versus fall larval connectivity and dispersal highlight the need for a greater understanding of spawning dynamics of the region. This study confirms that the Spratly Islands are likely an important source of larvae for the South China Sea and Coral Triangle region.     

Microhabitat characteristics of <Emphasis Type="Italic">Stegastes planifrons</Emphasis> and <Emphasis Type="Italic">S. adustus</Emphasis> territories

Stegastes adustus and Stegastes planifrons are two species of damselfishes commonly found in the Caribbean. These territorial fishes have been widely studied due to their major ecological role on coral reef in controlling the growth of macroalgae that compete with corals for space and, inversely, on their deleterious role in destroying coral tissues to impulse the development of algae. However, few studies were conducted on the biotic and abiotic components of their territories. In the present study, territory size and surfaces of benthic components (macroalgae, algal turf, massive corals, branching corals, Milleporidae, sponges, sand and rubbles) were estimated for the two species at two contrasted sites. At Ilet Pigeon site (IP), the two damselfishes were found at different depth and exhibited different territory sizes. S. adustus defended a larger territory characterized by massive corals, sand and Milleporidae, while S. planifrons territories were smaller, deeper and characterized by branching corals, sponges and rubble. At Passe-à-Colas site (PC), the two fish species coexisted in the same depth range and defended territories of similar size. Their territories presented higher proportions of macroalgae, but smaller surfaces of Milleporidae than at IP. At PC, the main difference between the two species was a higher surface of massive corals inside S. planifrons territories than S. adustus territories. Differences in microhabitat characteristics between the two Stegastes seemed mostly site related. This resulted from the high plasticity of two species, allowing them to persist on Caribbean coral reefs after the decline of most branching acroporids, their former favorite habitats.     

Mesophotic reef fish assemblages of the remote St. Peter and St. Paul’s Archipelago,Mid-Atlantic Ridge,Brazil

Mesophotic reef fish assemblages (30–90 m depth) of the small and remote St. Peter and St. Paul’s Archipelago (SPSPA), Mid-Atlantic Ridge, Brazil, were characterized using remotely operated vehicles. Ordination analyses identified distinct fish assemblages in the upper (30–50 m) and lower (50–90 m) mesophotic zones, the former characterized by high abundances of species that are also abundant at euphotic reefs (Caranx lugubris, Melichthys niger, Stegastes sanctipauli and Chromis multilineata) and the latter dominated by two mesophotic specialists (Prognathodes obliquus and Chromis enchrysura). Planktivores dominated fish assemblages, particularly in the upper mesophotic zone, possibly due to a greater availability of zooplankton coming from the colder Equatorial Undercurrent in mesophotic depths of the SPSPA. Turf algae, fleshy macroalgae and scleractinian corals dominated benthic assemblages between 30 and 40 m depth, while bryozoans, black corals and sponges dominated between 40 and 90 m depth. Canonical correspondence analysis explained 74 % of the relationship between environmental characteristics (depth, benthic cover and complexity) and structure of fish assemblages, with depth as the most important independent variable. Juveniles of Bodianus insularis and adults of P. obliquus and C. enchrysura were clearly associated with branching black corals (Tanacetipathes spp.), suggesting that black corals play key ecological roles in lower mesophotic reefs of the SPSPA. Results from this study add to the global database about mesophotic reef ecosystems (MREs) and provide a baseline for future evaluations of possible anthropogenic and natural disturbances on MREs of the SPSPA.     

Changes in gametogenesis and fecundity of acroporid corals that were exposed to elevated nitrogen and phosphorus during the ENCORE experiment

Colonies of two scleractinian reef coral species, Acropora longicyathus and Acropora aspera were transplanted into patch reefs at One Tree Reef, Great Barrier Reef, Australia as part of the ENCORE experiment. These corals and colonies of A. aspera which were naturally present in the patch reefs were exposed to four treatments over two years: controls with normal seawater, elevated levels of nitrogen only, phosphorus only, or nitrogen plus phosphorus. These corals were sampled and used to determine whether gametogenic cycles and fecundity were affected by nutrient enrichment. Acropora longicyathus had a single annual gametogenic cycle. Corals exposed to elevated nitrogen produced significantly smaller and fewer eggs and contained less testes material than those which were not exposed to nitrogen. Exposure to elevated phosphorus only resulted in corals producing more but smaller eggs, and more testes material. Egg numbers of colonies from other treatments decreased as the gametogenic cycles continued, but those of the phosphorus colonies showed almost no reduction in egg numbers between the early and late stages of the gametogenic cycles. These results have important management implications for coral reefs as they demonstrate that small increases in concentrations of nitrogen and phosphorus can have severe effects on reproductive activity in these species of scleractinian corals.     

Protection of Coral Larvae from Thermally Induced Oxidative Stress by Redox Nanoparticles

Coral reefs are one of the most biologically diverse and economically important ecosystems on earth. However, the destruction of coral reefs has been reported worldwide owing to rising seawater temperature associated with global warming. In this study, we investigated the potential of a redox nanoparticle (RNP^O) to scavenge reactive oxygen species (ROS), which are overproduced under heat stress and play a crucial role in causing coral mortality. When reef-building coral (Acropora tenuis) larvae, without algal symbionts, were exposed to thermal stress at 33 °C, RNP^O treatment significantly increased the survival rate. Proteome analysis of coral larvae was performed using nano-liquid chromatography-tandem mass spectrometry for the first time. The results revealed that several proteins related to ROS-induced oxidative stress were specifically identified in A. tenuis larvae without RNP^O treatment, whereas these proteins were absent in RNP^O-treated larvae, which suggested that RNP^O effectively scavenged ROS from A. tenuis larvae. Results from this study indicate that RNP^O treatment can reduce ROS in aposymbiotic coral larvae and would be a promising approach for protecting corals from thermal stress.     

Large-amplitude internal waves sustain coral health during thermal stress

Ocean warming is a major threat for coral reefs causing widespread coral bleaching and mortality. Potential refugia are thus crucial for coral survival. Exposure to large-amplitude internal waves (LAIW) mitigated heat stress and ensured coral survival and recovery during and after an extreme heat anomaly. The physiological status of two common corals, Porites lutea and Pocillopora meandrina, was monitored in host and symbiont traits, in response to LAIW-exposure throughout the unprecedented 2010 heat anomaly in the Andaman Sea. LAIW-exposed corals of both species survived and recovered, while LAIW-sheltered corals suffered partial and total mortality in P. lutea and P. meandrina, respectively. LAIW are ubiquitous in the tropics and potentially generate coral refuge areas. As thermal stress to corals is expected to increase in a warming ocean, the mechanisms linking coral bleaching to ocean dynamics will be crucial to predict coral survival on a warming planet.     

Temporal consistency in background mortality of four dominant coral taxa along Australia’s Great Barrier Reef

Studies on the population and community dynamics of scleractinian corals typically focus on catastrophic mortality associated with acute disturbances (e.g., coral bleaching and outbreaks of crown-of-thorns starfish), though corals are subject to high levels of background mortality and injuries caused by routine and chronic processes. This study quantified prevalence (proportion of colonies with injuries) and severity (areal extent of injuries on individual colonies) of background mortality and injuries for four common coral taxa (massive Porites, encrusting Montipora, Acropora hyacinthus and branching Pocillopora) on the Great Barrier Reef, Australia. Sampling was conducted over three consecutive years during which there were no major acute disturbances. A total of 2276 adult colonies were surveyed across 27 sites, within nine reefs and three distinct latitudinal sectors. The prevalence of injuries was very high (>83%) across all four taxa, but highest for Porites (91%) and Montipora (85%). For these taxa (Montipora and Pocillopora), there was also significant temporal and spatial variation in prevalence of partial mortality. The severity of injuries ranged from 3% to more than 80% and varied among coral taxa, but was fairly constant spatially and temporally. This shows that some injuries have considerable longevity and that corals may invest relatively little in regenerating tissue over sites of previous injuries. Inter-colony variation in the severity of injury also had no apparent effect on the realized growth of individual colonies, suggesting that energy diverted to regeneration has a limited bearing on overall energetic allocation, or impacts on other life-history processes (e.g., reproduction) rather than growth. Establishing background levels of injury and regeneration is important for understanding energy investment and life-history consequences for reef-building corals as well as for predicting susceptibility to, and capacity to recover from, acute disturbances.     

Reproductive biology and management of two commercially important groupers in the SW Atlantic

The reproductive biology of Epinephelus morio (red grouper) and Mycteroperca bonaci (black grouper) were evaluated based on 533 specimens collected from artisanal fisheries landings in the Abrolhos Bank, Brazil, between May 2005 and September 2012. Sex ratio for the black grouper was 1:14 (n = 155 females and 11 males; 26.1–147 cm TL) and 1:10 for the red grouper (n = 334 females and 33 males; 15.0–96.0 cm TL). For both species, highest values of the gonadosomatic index (GSI) for females were recorded between July and October, indicating spawning during the austral winter. The length at first maturity (L₅₀) for females was estimated at 62.0 and 47.0 cm TL for the black and red grouper, respectively. Batch fecundity based on TL and TW ranged from 2 to 15.4^?10⁶ and 1.5 to 13.7^?10⁶ for the black and red grouper, respectively. Interviews with experienced fishers revealed that spawning seasons of both groupers are largely unrecognized. Results demonstrate a positive relationship between GSI peaks, lower temperatures and stronger winds. The information provided herein may help decision-making regarding fisheries management and conservation for E. morio and M. bonaci at various levels of governance in the Abrolhos Bank, the region with the largest and richest coralline reefs in the South Atlantic.     

Into the depth of population genetics: pattern of structuring in mesophotic red coral populations

Deep-sea reef-building corals are among the most conspicuous invertebrates inhabiting the hard-bottom habitats worldwide and are particularly susceptible to human threats. The precious red coral (Corallium rubrum, L. 1758) has a wide bathymetric distribution, from shallow up to 800 m depth, and represents a key species in the Mediterranean mesophotic reefs. Several studies have investigated genetic variability in shallow-water red coral populations, while geographic patterns in mesophotic habitats are largely unknown. This study investigated genetic variability of C. rubrum populations dwelling between 55 and 120 m depth, from the Ligurian to the Ionian Sea along about 1500 km of coastline. A total of 18 deep rocky banks were sampled. Colonies were analyzed by means of a set of microsatellite loci and the putative control region of the mitochondrial DNA. Collected data were compared with previous studies. Both types of molecular markers showed high genetic similarity between populations within the northern (Ligurian Sea and Tuscan Archipelago) and the southern (Tyrrhenian and Ionian seas) study areas. Variability in habitat features between the sampling sites did not affect the genetic variability of the populations. Conversely, the patchy distribution of suitable habitats affected populations’ connectivity within and among deep coral banks. Based on these results and due to the emphasis on red coral protection in the Mediterranean Sea by international institutions, red coral could be promoted as a ‘focal species’ to develop management plans for the conservation of deep coralligenous reefs, a reservoir of marine biodiversity.     

Influence of the coral reef assemblages on the spatial distribution of echinoderms in a gradient of human impacts along the tropical Mexican Pacific

Fourteen species of echinoderms and their relationships to the benthic structure of the coral reefs were assessed at 27 sites—with different levels of human disturbances—along the coast of the Mexican Central Pacific. Diadema mexicanum and Phataria unifascialis were the most abundant species. The spatial variation of the echinoderm assemblages showed that D. mexicanum, Eucidaris thouarsii, P. unifascialis, Centrostephanus coronatus, Toxopneustes roseus, Holothuria fuscocinerea, Cucumaria flamma, and Echinometra vanbrunti accounted for the dissimilarities among the sites. The spatial variation among the sites was mainly explained by the cover of the hard corals (Porites, Pocillopora, Pavona, Psammocora), different macroalgae species (turf, encrusting calcareous algae, articulated calcareous algae, fleshy macroalgae), sponges, bryozoans, rocky, coral rubble, sand, soft corals (hydrocorals and octocorals), Tubastrea coccinea coral, Balanus spp., and water depth. The coverage of Porites, Pavona, and Pocillopora corals, soft coral, rock, and Balanos shows a positive relationship with the sampling sites included within the natural protected area with low human disturbances. Contrary, fleshy macroalgae, sponges, and soft coral show a positive relationship with higher disturbance sites. The results presented here show the importance of protecting the structural heterogeneity of coral reef habitats because it is a significant factor for the distribution of echinoderm species and can contribute to the design of conservation programs for the coral reef ecosystem.     

Microbiome variation in corals with distinct depth distribution ranges across a shallow–mesophotic gradient (15–85 m)

Mesophotic coral ecosystems (MCEs) are generally poorly studied, and our knowledge of lower MCEs (below 60 m depth) is largely limited to visual surveys. Here, we provide a first detailed assessment of the prokaryotic community associated with scleractinian corals over a depth gradient to the lower mesophotic realm (15–85 m). Specimens of three Caribbean coral species exhibiting differences in their depth distribution ranges (Agaricia grahamae, Madracis pharensis and Stephanocoenia intersepta) were collected with a manned submersible on the island of Curaçao, and their prokaryotic communities assessed using 16S rRNA gene sequencing analysis. Corals with narrower depth distribution ranges (depth-specialists) were associated with a stable prokaryotic community, whereas corals with a broader niche range (depth-generalists) revealed a higher variability in their prokaryotic community. The observed depth effects match previously described patterns in Symbiodinium depth zonation. This highlights the contribution of structured microbial communities over depth to the coral’s ability to colonize a broader depth range.     

Age,growth and reproduction of the Iranian goby, <Emphasis Type="Italic">Ponticola iranicus</Emphasis> (Perciformes: Gobiidae) from the southern Caspian Sea basin

To estimate reproductive biology of Ponticola iranicus Vasil’eva et al., 2015, in Sefidroud River in the southern Caspian Sea basin age, sex ratio, fecundity, oocytes diameter, gonadosomatic, modified gonadosomatic and dobriyal indices were measured. Regression analyses were used to find relations between fecundity and fish size (length and weight), gonad weight, and age. Mature females and males were longer than 46 and 49 mm in total length, +1 in age. The Von Bertalanffy growth models were L _∞ = 23.00 cm (TL), K = 0.21, t ₀ =–0.25 for all specimens. Sex ratio was 1: 1.32 in favor of males. The average egg diameter was 0.96 mm. The spawning period extended from late February to late April. The averages of absolute and relative fecundity were calculated as 536.9 and 81.3, respectively. The absolute fecundity was significantly related to the body length and weight, as well as gonad weight.     

Long-term dynamics of the brown macroalga <Emphasis Type="Italic">Lobophora variegata</Emphasis> on deep reefs in Curaçao

Lobophora variegata occurs in the eulittoral zone and in deep water on coral reefs in Curaçao. An analysis of the long-term (1979–2006) changes in the vertical distribution of the macroalga in permanent quadrats indicated a significant increase in cover of the deepwater community. In 1998, Lobophora covered 1 and 5% of the quadrats at 20 and 30 m, respectively. By 2006, these values had risen to 25 and 18%, precipitating a shift in abundance of corals and macroalgae at both depths. This increase coincided with losses in coral cover, possibly linked to bleaching, disease and storm-related mortality in deep water plating Agaricia corals. In contrast, macroalgae remained relatively rare (<6% cover) on shallower (10 m) and deeper (40 m) reefs despite declines in coral cover also occurring at these depths, illustrating the depth-dependent dynamics of coral reefs. Several hypotheses are suggested to explain these changes.     

Coral reproduction in the world��s warmest reefs: southern Persian Gulf (Dubai, United Arab Emirates)

Despite extensive research on coral reproduction from numerous geographic locations, there remains limited knowledge within the Persian Gulf. Given that corals in the Persian Gulf exist in one of the most stressful environments for reef corals, with annual variations in sea surface temperature (SST) of 12°C and maximum summer mean SSTs of 36°C, understanding coral reproductive biology in the Gulf may provide clues as to how corals may cope with global warming. In this study, we examined six locally common coral species on two shallow reef sites in Dubai, United Arab Emirates (UAE), in 2008 and 2009 to investigate the patterns of reproduction, in particular the timing and synchrony of spawning. In total, 71% colonies in April 2008 and 63% colonies in April 2009 contained mature oocytes. However, the presence of mature gametes in May indicated that spawning was potentially split between April and May in all species. These results demonstrate that coral reproduction patterns within this region are highly seasonal and that multi-species spawning synchrony is highly probable. Acropora downingi, Cyphastrea microphthalma and Platygyra daedalea were all hermaphroditic broadcast spawners with a single annual gametogenic cycle. Furthermore, fecundity and mature oocyte sizes were comparable to those in other regions. We conclude that the reproductive biology of corals in the southern Persian Gulf is similar to other regions, indicating that these species have adapted to the extreme environmental conditions in the southern Persian Gulf.     

The functional importance of <Emphasis Type="Italic">Acropora austera</Emphasis> as nursery areas for juvenile reef fish on South African coral reefs

Many coral reef fish species use mangrove and seagrass beds as nursery areas. However, in certain regions, the absence or scarcity of such habitats suggests that juvenile coral reef fish may be seeking refuge elsewhere. The underlying biogenic substratum of most coral reefs is structurally complex and provides many types of refuge. However, on young or subtropical coral reefs, species may be more reliant on the living coral layer as nursery areas. Such is the case on the high-latitude coral reefs of South Africa where the coral communities consist of a thin veneer of coral overlaying late Pleistocene bedrock. Thus, the morphology of coral species may be a major determinant in the availability of refuge space. Acropora austera is a branching species that forms large patches with high structural complexity. Associated with these patches is a diverse community of fish species, particularly juveniles. Over the past decade, several large (>100 m²) A. austera patches at Sodwana Bay have been diminishing for unknown reasons and there is little evidence of their replacement or regrowth. Seven patches of A. austera (AP) and non-A. austera (NAP) were selected and monitored for 12 months using visual surveys to investigate the importance of AP as refugia and nursery areas. There were significant differences in fish communities between AP and NAP habitats. In total, 110 species were recorded within the patches compared to 101 species outside the patches. Labrids and pomacentrids were the dominant species in the AP habitats, while juvenile scarids, acanthurids, chaetodons and serranids were also abundant. The diversity and abundance of fish species increased significantly with AP size. As the most structurally complex coral species on the reefs, the loss of APs may have significant implications for the recruitment and survival of certain fish species.     

The effect of seasonal temperature extremes on sediment rejection in three scleractinian coral species

Sedimentation from resuspension following storm surge is a natural occurrence on coral reefs, and scleractinian corals have adapted to effectively reject sediment. However, it is unclear whether the physical ability to reject sedimentation is affected during seasonal temperature extremes. We acclimated three coral species (Montipora aequituberculata, Lobophyllia corymbosa and Fungia fungites), with different active shedding mechanisms, to three temperature treatments (winter minimum, summer maximum and mean). Corals were then exposed to a sediment rejection experiment in which we measured clearance rates and tissue inflation cycles associated with the clearance of sediment. Temperature impacted clearing rates of M. aequituberculata, which exhibited significantly faster sediment rejection under winter temperatures. Fungia fungites, on the other hand, exhibited significantly higher tissue inflation rates under summer temperatures. Although limited in scope, this study demonstrates that temperature can have a strong effect on the response of corals to sedimentation.