Organellar genomic characterization of Anunuuluaehu liula representing a new genus and species of Phyllophoraceae (Gigartinales, Rhodophyta) from the mesophotic zone of Hawai'i.

Over the last 2 decades, routine collections in the Hawaiian Archipelago have expanded to mesophotic reefs, leading to the discovery of a new red algal genus and species, here described as Anunuuluaehu liula gen. et sp. nov. This study provides a detailed genus and species description and characterizes chloroplast and mitochondrial organellar genomes. The new genus, Anunuuluaehu, shares many characteristics with the family Phyllophoraceae and shows close similarities to Archestennogramma and Stenogramma, including habit morphology, nemathecia forming proliferations at the outer cortex with terminal chains of tetrasporangia, and carposporophytes with multi-layered pericarps. The single species in this genus exhibits distinctive features within the Phyllophoraceae: the presence of single-layer construction of large medullary cells and the development of long, tubular gonimoblastic filaments. Multi-gene phylogenetic analyses confirmed it as a unique, monophyletic lineage within the family. Cis-splicing genes, interrupted by intron-encoded proteins within group II introns, are present in both the chloroplast and mitochondrial genomes of A. liula. Notably, a specific region of the coxI group II intron exhibits similarity to fungal introns. Anunuuluaehu liula is presumed to be endemic to the Hawaiian Archipelago and thus far is known to live solely at mesophotic depths from Holaniku to Kaho'olawe ranging from 54 to 201 m, which is the deepest collection record of any representative in the family. Overall, this study enhances our understanding of the genomic and taxonomic complexities of red algae in mesophotic habitats, emphasizing the significance of continued research in this area to uncover further insights into evolutionary processes and biogeographic patterns.

Characterization of macroalgal-associated microbial communities from shallow to mesophotic depths at Manawai, Papahanaumokuakea Marine National Monument, Hawai'i.

The Papahanaumokuakea Marine National Monument, Hawai'i, is one of the most isolated and protected archipelagos in the world, making it a natural laboratory to examine macroalgal-microbial diversity because of limited direct anthropogenic impacts. We collected the most abundant macroalgae from nine sites ranging from shallow subtidal (1.5 m) to mesophotic (75 m) depths around Manawai (Pearl and Hermes Atoll). We characterized the macroalgal bacterial communities via high-throughput amplicon sequencing and compared the influence of host phylum, species, site, and depth on these relationships at a single atoll. Ochrophyta species had the lowest bacterial diversity compared to Chlorophyta and Rhodophyta. Site and/or depth may influence the microbial community structure associated with Microdictyon setchellianum, indicating a possible disconnect of these microbial communities among habitats. Chondria tumulosa, a cryptogenic species with invasive traits, differed in associated microbiota compared to the native Laurencia galtsoffii, an alga from the same family collected at the same site and depth. While there was overlap of bacterial communities across sites for some algal species, the majority had minimal macroalgal-microbial community connectivity across Manawai. This mesophotic system, therefore, did not appear to be refugia for shallow water coral reefs at microscopic scales. Additional studies are required to identify other significant influences on microbial community variation.

Complete chloroplast genome of Chondria tumulosa (Ceramiales, Rhodophyta), a recently described cryptogenic species with invasive traits from Papahanaumokuakea Marine National Monument, Hawai'i.

The complete chloroplast genome of Chondria tumulosa, a red alga from Manawai (Pearl and Hermes Atoll), Hawai'i, was determined and analyzed using next-generation sequencing and de novo assembly approaches. The chloroplast genome sequence of C. tumulosa was 172,617 bp and contained 231 genes, consisting of 197 protein-coding genes, 29 transfer RNA genes, three ribosomal RNA genes, one transfer-messenger RNA gene, one non-coding RNA gene, and one intron inserted into the trnM gene. The number of genes and genome structure was largely similar to other members of the family Rhodomelaceae. The phylogenomic analysis of 32 complete cpDNA from the red algal order Ceramiales showed that C. tumulosa is a distinct species within the Chondrieae tribe, and is a diverging early relative to the other three available Chondria chloroplast genomes.

3D assessment of a coral reef at Lalo Atoll reveals varying responses of habitat metrics following a catastrophic hurricane.

Extreme disturbances such as hurricanes can cause reductions in coral cover and three-dimensional (3D) structural complexity of coral reefs. We examined changes in structural complexity utilizing 3D reconstruction of a coral-reef site before and after Hurricane Walaka passed through Lalo of the Northwestern Hawaiian Islands. This event resulted in complete destruction of the coral-reef habitat, with dramatic changes in benthic cover from pre-hurricane tabulate coral to post-hurricane rubble. Rugosity and mean slope decreased after the hurricane, while structural complexity, captured by vector ruggedness measure (VRM), showed resolution-specific responses. This metric captured the structural complexity of rubble at a high raster resolution of 1 cm and that of tabulate coral at lower resolutions, resulting in decreases in mean VRM values at 2- and 4-cm resolutions but an increase at 1-cm resolution. Variability in profile and planform curvature was reduced after the hurricane due to a disappearance of extreme curvature values created by the tabulate coral after the hurricane. This study highlights the varying responses of habitat complexity metrics to the complete destruction of a coral reef and provides us with insights into how choices of habitat complexity metrics can affect quantitative assessments of 3D habitat structure.

Taxonomic determination of the cryptogenic red alga, Chondria tumulosa sp. nov., (Rhodomelaceae, Rhodophyta) from Papahanaumokuakea Marine National Monument, Hawai'i, USA: A new species displaying invasive characteristics.

Survey cruises by the National Oceanic and Atmospheric Administration (NOAA) in 2016 and 2019 yielded specimens of an undetermined red alga that rapidly attained alarming levels of benthic coverage at Pearl and Hermes Atoll, Papahanaumokuakea Marine National Monument, Hawai'i. By 2019 the seaweed had covered large expanses on the northeast side of the atoll with mat-like, extensive growth of entangled thalli. Specimens were analyzed using light microscopy and molecular analysis, and were compared to morphological descriptions in the literature for closely related taxa. Light microscopy demonstrated that the specimens likely belonged to the rhodomelacean genus Chondria, yet comparisons to taxonomic literature revealed no morphological match. DNA sequence analyses of the mitochondrial COI barcode marker, the plastidial rbcL gene, and the nuclear SSU gene confirmed its genus-level placement and demonstrated that this alga was unique compared to all other available sequences. Based on these data, this cryptogenic seaweed is here proposed as a new species: Chondria tumulosa A.R.Sherwood & J.M.Huisman sp. nov. Chondria tumulosa is distinct from all other species of Chondria based on its large, robust thalli, a mat-forming tendency, large axial diameter in mature branches (which decreases in diameter with subsequent orders of branching), terete axes, and bluntly rounded apices. Although C. tumulosa does not meet the criteria for the definition of an invasive species given that it has not been confirmed as introduced to Pearl and Hermes Atoll, this seaweed is not closely related to any known Hawaiian native species and is of particular concern given its sudden appearance and rapid increase in abundance in the Papahanaumokuakea Marine National Monument; an uninhabited, remote, and pristine island chain to the northwest of the Main Hawaiian Islands.

Effects of SCUBA bubbles on counts of roving piscivores in a large remote marine protected area.

This study examined the effects of SCUBA bubbles on fish counts in underwater visual surveys conducted in the Papahanaumokuakea Marine National Monument (PMNM). Specifically, paired fish surveys were conducted at each survey site, utilizing two different gear types: open-circuit SCUBA (OC) and closed-circuit rebreather (CCR). Bubble exhaust released from the OC equipment is a potential source of bias for in-situ fish observations, as the associated audio and visual disturbances could either attract or repel fishes depending on whether their behavior is more driven by curiosity or caution. The study area, is a large (~1.5 million km2) and extremely remote marine protected area in which the response of coral reef fishes to divers represent natural behavior of naive fishes with little or no previous contact with humans. Historically, surveys conducted on OC in this area have shown an abundance of large roving piscivores and this study set out to determine the extant, if any, the audible and visual disturbances of OC bubbles have. The species typically seen in these prior surveys were Caranx ignobilis, Caranx melampygus, Aprion virescens, and a couple of species of sharks. We found differences in counts for some roving piscivores, including significantly more jacks observed on OC than CCR (Caranx ignobilis 57% more, and Caranx melampygus 113% more). Instance of first encounter, i.e. the time when a fish was first observed during a survey, also varied for some species. Higher numbers of Aprion virescens (p = 0.04), and C. melampygus (p = <0.001) were observed in the first 5-minutes of counts by divers on OC (i.e. when they were using breathing apparatus that produced noises that could be heard over long distances). Although not the focus of the study, we also assessed differences between OC and CCR counts for other groups of fishes. Estimated abundance of benthic damselfish was higher on OC than CCR, and counts of butterflyfish were lower on OC; but there were no significant differences for the other groups considered. This is an important control study that documents the natural responses of coral reef fishes to SCUBA bubbles generated by in-situ surveys.

Long-term monitoring of coral reef fish assemblages in the Western central pacific.

Throughout the tropics, coral reef ecosystems, which are critically important to people, have been greatly altered by humans. Differentiating human impacts from natural drivers of ecosystem state is essential to effective management. Here we present a dataset from a large-scale monitoring program that surveys coral reef fish assemblages and habitats encompassing the bulk of the US-affiliated tropical Pacific, and spanning wide gradients in both natural drivers and human impact. Currently, this includes >5,500 surveys from 39 islands and atolls in Hawaii (including the main and Northwestern Hawaiian Islands) and affiliated geo-political regions of American Samoa, the Commonwealth of the Northern Mariana Islands, Guam, and the Pacific Remote Islands Areas. The dataset spans 2010-2017, during which time, each region was visited at least every three years, and ~500-1,000 surveys performed annually. This standardised dataset is a powerful resource that can be used to understand how human, environmental and oceanographic conditions influence coral reef fish community structure and function, providing a basis for research to support effective management outcomes.

Mass coral bleaching due to unprecedented marine heatwave in Papahanaumokuakea Marine National Monument (Northwestern Hawaiian Islands).

2014 marked the sixth and most widespread mass bleaching event reported in the Northwestern Hawaiian Islands, home to the Papahanaumokuakea Marine National Monument (PMNM), the world's second largest marine reserve. This event was associated with an unusual basin-scale warming in the North Pacific Ocean, with an unprecedented peak intensity of around 20°C-weeks of cumulative heat stress at Lisianksi Island. In situ bleaching surveys and satellite data were used to evaluate the relative importance of potential drivers of bleaching patterns in 2014, assess the subsequent morality and its effects on coral communities and 3D complexity, test for signs of regional acclimation, and investigate long-term change in heat stress in PMNM. Surveys conducted at four island/atoll (French Frigate Shoals, Lisianski Island, Pearl and Hermes Atoll, and Midway Atoll) showed that in 2014, percent bleaching varied considerably between islands/atolls and habitats (back reef/fore reef and depth), and was up to 91% in shallow habitats at Lisianski. The percent bleaching during the 2014 event was best explained by a combination of duration of heat stress measured by Coral Reef Watch's satellite Degree Heating Week, relative community susceptibility (bleaching susceptibility score of each taxon * the taxon's abundance relative to the total number of colonies), depth and region. Mean coral cover at permanent Lisianski monitoring sites decreased by 68% due to severe losses of Montipora dilatata complex, resulting in rapid reductions in habitat complexity. Spatial distribution of the 2014 bleaching was significantly different from the 2002 and 2004 bleaching events likely due to a combination of differences in heat stress and local acclimatization. Historical satellite data demonstrated heat stress in 2014 was unlike any previous event and that the exposure of corals to the bleaching-level heat stress has increased significantly in the northern PMNM since 1982, highlighting the increasing threat of climate change to reefs.

Use of multivariate control charts to assess the status of reef fish assemblages in the Northwestern Hawaiian Islands.

A distance-based multivariate control chart is a useful tool for ecological monitoring to detect changes in biological community resulting from natural or anthropogenic disturbances at permanent monitoring sites. It is based on a matrix of any distances or dissimilarities among observations obtained from species composition and abundance data, and bootstrapping techniques are used to set upper confidence bounds that trigger an alarm for further investigations. We extended the use of multivariate control charts to stratified random sampling and analyzed reef fish monitoring data collected annually on shallow (≤30 m) reefs across the Northwestern Hawaiian Islands (NWHI), part of the Papahanaumokuakea Marine National Monument. Fish assemblages in the NWHI were mostly stable, with exceptions in the south region (Nihoa, Mokumanamana and French Frigate Shoals) in 2012 and 2015 where changes in the assemblage structure exceeded the upper confidence bounds of multivariate control charts. However, these were due to changes in relative abundances of native species, and potentially related to the small numbers of survey sites and relatively low coral covers at the sites, particularly in 2015. The present study showed that multivariate control charts can be used to evaluate the status of biological communities in a very large protected area. Future monitoring of fish assemblages in the Papahanaumokuakea Marine National Monument should be accompanied by specific habitat or environmental variables that are related to potential threats to its shallow-water ecosystems. This should allow for more detailed investigations into potential causes and mechanisms of changes in fish assemblages when a multivariate control chart triggers an alarm.

Comparison of Reef Fish Survey Data Gathered by Open and Closed Circuit SCUBA Divers Reveals Differences in Areas With Higher Fishing Pressure.

Visual survey by divers using open-circuit (OC) SCUBA is the most widely used approach to survey coral reef fishes. Therefore, it is important to quantify sources of bias in OC surveys, such as the possibility that avoidance of OC divers by fishes can lead to undercounting in areas where targeted species have come to associate divers with a risk of being speared. One potential way to reduce diver avoidance is to utilize closed circuit rebreathers (CCRs), which do not produce the noise and bubbles that are a major source of disturbance associated with OC diving. For this study, we conducted 66 paired OC and CCR fish surveys in the Main Hawaiian Islands at locations with relatively high, moderate, and light fishing pressure. We found no significant differences in biomass estimates between OC and CCR surveys when data were pooled across all sites, however there were differences at the most heavily fished location, Oahu. There, biomass estimates from OC divers were significantly lower for several targeted fish groups, including surgeonfishes, targeted wrasses, and snappers, as well as for all targeted fishes combined, with mean OC biomass between 32 and 68% of mean CCR biomass. There were no clear differences between OC and CCR biomass estimates for these groups at sites with moderate or low fishing pressure, or at any location for other targeted fish groups, including groupers, parrotfishes, and goatfishes. Bias associated with avoidance of OC divers at heavily fished locations could be substantially reduced, or at least calibrated for, by utilization of CCR. In addition to being affected by fishing pressure, the extent to which avoidance of OC divers is problematic for visual surveys varies greatly among taxa, and is likely to be highly influenced by the survey methodology and dimensions used.

A comprehensive investigation of mesophotic coral ecosystems in the Hawaiian Archipelago.

Although the existence of coral-reef habitats at depths to 165 m in tropical regions has been known for decades, the richness, diversity, and ecological importance of mesophotic coral ecosystems (MCEs) has only recently become widely acknowledged. During an interdisciplinary effort spanning more than two decades, we characterized the most expansive MCEs ever recorded, with vast macroalgal communities and areas of 100% coral cover between depths of 50-90 m extending for tens of km2 in the Hawaiian Archipelago. We used a variety of sensors and techniques to establish geophysical characteristics. Biodiversity patterns were established from visual and video observations and collected specimens obtained from submersible, remotely operated vehicles and mixed-gas SCUBA and rebreather dives. Population dynamics based on age, growth and fecundity estimates of selected fish species were obtained from laser-videogrammetry, specimens, and otolith preparations. Trophic dynamics were determined using carbon and nitrogen stable isotopic analyses on more than 750 reef fishes. MCEs are associated with clear water and suitable substrate. In comparison to shallow reefs in the Hawaiian Archipelago, inhabitants of MCEs have lower total diversity, harbor new and unique species, and have higher rates of endemism in fishes. Fish species present in shallow and mesophotic depths have similar population and trophic (except benthic invertivores) structures and high genetic connectivity with lower fecundity at mesophotic depths. MCEs in Hawai'i are widespread but associated with specific geophysical characteristics. High genetic, ecological and trophic connectivity establish the potential for MCEs to serve as refugia for some species, but our results question the premise that MCEs are more resilient than shallow reefs. We found that endemism within MCEs increases with depth, and our results do not support suggestions of a global faunal break at 60 m. Our findings enhance the scientific foundations for conservation and management of MCEs, and provide a template for future interdisciplinary research on MCEs worldwide.

Prognathodes basabei, a new species of butterflyfish (Perciformes, Chaetodontidae) from the Hawaiian Archipelago.

A new species of the butterflyfish genus Prognathodes is described from specimens collected at a depth of 55-61 m off Pearl and Hermes Atoll, Northwestern Hawaiian Islands. This species has been observed by mixed-gas divers and from submersibles at depths ranging from 45-187 m throughout the Hawaiian Archipelago, with shallower sightings in the Northwestern Hawaiian Islands and deeper in the Main Hawaiian Islands. It is similar to Prognathodes guezei (Maugé and Bauchot 1976) from the western Indian Ocean, and at least one other undescribed species of Prognathodes from Palau, differing from these species in the number of soft dorsal-fin rays, size of head, and body depth. There are also differences in the life color, and a substantial genetic difference from the Palauan species (d ¼ .08 in mtDNA cytochrome oxidase I).

Structure of Mesophotic Reef Fish Assemblages in the Northwestern Hawaiian Islands.

Mesophotic coral ecosystems (MCEs) support diverse communities of marine organisms with changes in community structure occurring along a depth gradient. In recent years, MCEs have gained attention due to their depths that provide protection from natural and anthropogenic stressors and their relative stability over evolutionary time periods, yet ecological structures of fish assemblages in MCEs remain largely un-documented. Here, we investigated composition and trophic structure of reef fish assemblages in the Northwestern Hawaiian Islands (NWHI) along a depth gradient from 1 to 67 m. The structure of reef fish assemblages as a whole showed a clear gradient from shallow to mesophotic depths. Fish assemblages at mesophotic depths had higher total densities than those in shallower waters, and were characterized by relatively high densities of planktivores and invertivores and relatively low densities of herbivores. Fishes that typified assemblages at mesophotic depths included six species that are endemic to the Hawaiian Islands. The present study showed that mesophotic reefs in the NWHI support unique assemblages of fish that are characterized by high endemism and relatively high densities of planktivores. Our findings underscore the ecological importance of these undersurveyed ecosystems and warrant further studies of MCEs.

Ecological and morphological traits predict depth-generalist fishes on coral reefs.

Ecological communities that occupy similar habitats may exhibit functional convergence despite significant geographical distances and taxonomic dissimilarity. On coral reefs, steep gradients in key environmental variables (e.g. light and wave energy) restrict some species to shallow depths. We show that depth-generalist reef fishes are correlated with two species-level traits: caudal fin aspect ratio and diet. Fishes with high aspect ratio (lunate) caudal fins produce weaker vortices in the water column while swimming, and we propose that 'silent swimming' reduces the likelihood of detection and provides an advantage on deeper reefs with lower light irradiance and water motion. Significant differences in depth preference among trophic guilds reflect variations in the availability of different food sources along a depth gradient. The significance of these two traits across three geographically and taxonomically distinct assemblages suggests that deep-water habitats exert a strong environmental filter on coral reef-fish assemblages.

Tosanoides obama, a new basslet (Perciformes, Percoidei, Serranidae) from deep coral reefs in the Northwestern Hawaiian Islands.

The new species Tosanoides obama is described from two specimens collected at a depth of 90-92 m off Kure Atoll and Pearl and Hermes Atoll, Northwestern Hawaiian Islands. It differs from the other two species of this genus in life color and in certain morphological characters, such as number of pored lateral-line scales, pectoral-fin rays, snout length, anterior three dorsal-fin spine lengths, dorsal-fin profile, and other characters. There are also substantial genetic differences from the other two species of Tosanoides (d ≈ 0.10 in mtDNA cytochrome oxidase I). The species is presently known only from the Northwestern Hawaiian Islands within the Papahanaumokuakea Marine National Monument.

Acoustic communication in territorial butterflyfish: test of the sound production hypothesis.

Butterflyfishes are conspicuous members of coral reefs and well known for their visual displays during social interactions. Members of the genus Chaetodon have a unique peripheral arrangement of the anterior swim bladder that connects with the lateral line (the laterophysic connection) and in many species projects towards the inner ear. This morphology has lead to the proposal that the laterophysic connection and swim bladder system may be a specialized structure for the detection of sound. However, the relevant stimuli, receiver mechanisms and functions for these putative hearing structures were unknown because butterflyfishes were previously not recognized to produce sounds during natural behavior. We performed field experiments to test the hypothesis that Chaetodon produces sounds in natural social contexts. Acoustic and motor behaviors of the monogamous multiband butterflyfish, C. multicinctus, were evoked and recorded by placement of bottled fish into feeding territories of conspecific pairs. We demonstrate that territory defense includes the production of agonistic sounds and hydrodynamic stimuli that are associated with tail slap, jump, pelvic fin flick and dorsal-anal fin erection behaviors. In addition, grunt pulse trains were produced by bottled intruders and are tentatively interpreted to function as an alert call among pair mates. Acoustic behaviors include low frequency hydrodynamic pulses <100 Hz, sounds with peak energy from 100 Hz to 500 Hz, and a broadband high frequency click (peak frequency=3.6 kHz), which is produced only during the tail slap behavior. These results provide a biological framework for future studies to interpret the proximate function of the acoustico-lateralis sensory system, the evolution of the laterophysic mechanism and their relevance to butterflyfish social behavior.